我々は知識欲と行動力を持ち、ヒト・動物・環境の問題を解決するためにグローバルな活躍を目指す人材を求めています。当プログラムに参加し、地球規模でのOne Healthの実現に貢献しませんか？
プログレスの報告
「プログレス」とは、大学院生が自主的に企画・運営する研究討論会です。
1. Bongkot Soonthornsata (Unit of Risk Analysis and Management, CZC, D1)
"Transmission risks of major mosquito-borne diseases in Thailand"
Introduction
Mosquito-borne diseases are causing a substantial burden of mortality, morbidity and economic loss in many parts of the world including Thailand and its neighboring countries. Given the persistent of case notifications of dengue and chikungunya including a recent uncovered Zika cases in Thailand, in combination with the abundance and presence of their significant vectors, Aedes aegypti and Aedes albopictus in the entire country, high level of population movement along border areas and within the country, and changes in environment, indicating that transmission in endemic areas will likely be continued and it is possible that transmission potential in new risk areas where suitable vectors are present will be identified. The current research is thus to investigate transmission risks of dengue and other relevant diseases in Thailand by conducting entomological and epidemiological surveillances and all data collected will be statistically assessed. The study will be taken place at 13 sites throughout Thailand and all study sites intend to start conducting their first entomological surveillance in February 2016 and mosquito samples are expected to collect three times a year. Mosquito samples collected will be used for molecular detection and genotyping of pathogens in mosquitoes and/or for monitoring of currently used insecticide resistance status.
2. Tomoya Morita (Lab. of Internal Medicine, D2)
"Assessment of right heart function by echocardiography in dog models of acute pulmonary hypertension and dogs with heart disease."
Introduction
Right heart dysfunction has recently been shown to predict poor prognosis and symptom in patients with heart disease. Therefore, right heart function test attracts attention in human medicine. However, the quantitative assessment of right heart function using noninvasive method, such as echocardiography, is challenging because of the complex shape of right ventricular (RV). Tei index is an echocardiographic index of the global myocardial function, including systolic and diastolic performance. Tei index is unaffected by complex shape of RV, and it has been shown to predict the prognosis, severity of the human patients with heart disease. However, few reports are available on the relationship between Tei index and hemodynamic variables or prognosis in dogs. Therefore, we measured Tei index in dog models of acute RV pressure overload and client-owned dogs with heart disease. As a result, Tei index reflected hemodynamic deterioration and diastolic dysfunction in dog models of acute RV pressure overload, and it was a good predictor of death in dogs with heart disease. For the future, we`ll create dog models of chronic RV pressure overload, and validate the usefulness of Tei index in the assessment of right heart function.
3. Memi Muto (Lab. of Public Health, D2)
"Isolation and characterization of tick-borne encephalitis virus from Ixodes persulcatus in Mongolia in 2012."
Introduction
Tick-borne encephalitis virus (TBEV) is a zoonotic virus belonging to the genus Flavivirus, in the family Flaviviridae. The virus, which is endemic in Europe and northern parts of Asia, causes severe encephalitis. Tick-borne encephalitis (TBE) has been reported in Mongolia since the 1980s, but details about the biological characteristics of the endemic virus are lacking. In this study, 680 ticks (Ixodes persulcatus) were collected in Selenge aimag, northern Mongolia, in 2012. Nine Mongolian TBEV strains were isolated from tick homogenates. A sequence analysis of the envelope protein gene revealed that all isolates belonged to the Siberian subtype of TBEV. Two strains showed similar growth properties in cultured cells, but their virulence in mice differed. Whole genome sequencing revealed only thirteen amino acid differences between these Mongolian TBEV strains. Our results suggest that these naturally occurring amino acid mutations affected the pathogenicity of Mongolian TBEV. Our results may be an important platform for monitoring TBEV to evaluate the epidemiological risk in TBE endemic areas of Mongolia.
4. Wallaya Phongphaew (Division of Molecular Pathobiology, CZC, D2)
"Valosin containing protein plays an important role in West Nile virus infection"
Introduction
The Endoplasmic Reticulum is related to the genome replication and assembly steps of WNV life cycle. The Valosin containing protein (VCP/p97) is a one component of ER-associated degradation (ERAD) pathway involving in transportation of ubiquitinated protein to proteasome for degradation and the other cellular processes. Previous studies revealed that VCP also participates in the life cycle of several viruses such as Poliovirus and Sindbis virus. The present study demonstrates the role of the VCP in WNV infection. Inhibition of VCP by treatment with either Eeyarestatin I or 3,4-Methylenedioxy-β-nitrostyrene inhibits the WNV infection in a dose-dependent manner. Furthermore, the depletion of the endogenous VCP expression using siRNA significantly attenuates the WNV infection. Therefore, we conclude that the VCP is required for WNV infection. We also examine which step of WNV life cycle should be participated by VCP. The results suggest that VCP may play a role in the either WNV genome replication or protein synthesis.
5. Tapiwa Lundu (Laboratory of Public Health, D2)
"Studies on the ecology of Severe Fever with Thrombocytopenia Syndrome Virus and the analysis of the mechanisms of virion formation of SFTSV"
Introduction
1) A variety of animals are suggested to be reservoir hosts of severe fever with thrombocytopenia syndrome virus (SFTSV). In this study we screened 315 deer sera from Hokkaido (non-endemic area), 41 deer sera from Miyazaki (endemic area), 4 wild boar sera from Nagasaki and 910 rodent sera from six locations in the endemic area of Japan. All the sera from Hokkaido showed a negative result by ELISA (OD<0.1). Fourteen rodent sera had slightly higher OD values on ELISA (0.1
2) In the bunyavirus replication cycle, for assembly of virions to occur, nucleoprotein and glycoproteins must move to the same intracellular location. The Gn protein of members of the phlebovirus possesses a Golgi retention signal which assists in localisation of Gn and Gc to the Golgi. For NP to localise to the Golgi, it may need to interact with glycoprotein. In this study we hope to show that the SFTSV N and Gn proteins interact and thereby directing NP to the site of virus assembly.
1. Tatsunari Kondoh (Division of Global Epidemiology, CZC, D1)
"Host factors involved in the cell susceptibility to filovirus"
Introduction
1) Niemann-Pick C1 (NPC1), a protein involved in filovirus glycoprotein (GP)-mediated membrane fusion during the virus entry into cells, is believed to be a major determinant of cell susceptibility to filovirus infection. To analyze polymorphism of human NPC1s, we cloned NPC1 genes from permissive and non-permissive cell lines for filovirus infection and detected nonsynonymous polymorphism in these NPC1s. Three amino acid differences were found between NPC1s of permissive and non-permissive cell lines. We hypothesize that this polymorphism may affect efficiency of filovirus entry. In the future, we will generate cell lines expressing each NPC1 to assess the GP-dependent entry.
2) It has been reported that some non-retroviral RNA virus genes are integrated into vertebrate genomes. Recent studies suggest that expression of endogenously encoded bornavirus-like nucleoprotein confers resistance to the virus infection by inhibiting its replication in cells. Endogenous filovirus-like elements (EFLEs) have also been discovered in several mammalian genomes, however the role of EFLEs is not known. We constructed plasmid expressing an endogenous filovirus VP35-like element (mlEEL35) containing nearly full-length genes corresponding to the VP35 gene. We will express the mlEEL35 in cell culture and clarify whether it has a VP35-like function (i.e., inhibition of interferon response).
2. Rabin Kadariya (Lab. of Wildlife Biology and Medicine, D1)
"Genetic Variation, Ecology and Conservation of Asiatic Black Bears (Ursus thibetanus) in Annapurna Conservation Area of Nepal"
Introduction
The Asiatic black bear is endangered in mountain landscape of Nepal but has never been received for conservation importance. Retaliatory killing, poaching, climate change and habitat degradation have led to dwindling and scattered bear populations, however, no study on population genetic structure and ecology was conducted. Genetic variation may reduce in small breeding population due to inbreeding and genetic drift that impede adaptability of population to new environment. This situation is true in spatially isolated small bear population in Nepal. It may lead to extirpation of certain populations if timely conservation measures are not undertaken. This research explores genetic variation and seasonal diet preference of isolated bear population using non-invasive method.
During initial survey, 25 fecal and 10 hair samples were collected from bear habitat. DNA from fecal and hair samples was extracted using QIAamp mini-stool kit and DNA extractor FM kit (Wako) respectively. The highly variable region of mitochondrial DNA control region and nuclear DNA microsatellite will be sequenced for study of phylogenetic relationship and genetic variation respectively. The percent frequency of occurrence shows that wild fruits/nuts (85%) are major food followed grasses (43%) whereas small amount of mammal, insects and crop are also recorded in bear feces.
3. Suranji Wijekoon (Lab. of Veterinary Surgery, D2)
"Osteoclast precursor as cellular biomarker to assess relative protection against bone erosion in concurrent patellar luxation and cranial cruciate ligament rupture among dogs"
Introduction
Inflammatory joint disease characterized by abnormal synovial proliferation and destruction of articular cartilage and bone. Synovial membranes (SM) were collected from dogs associated with cranial cruciate ligament rupture (CCLr, n=5) and medial patella luxation (MPL, n=5). Samples were homogenized to extract RNA and another part was cultured in the presence of receptor-activator for nuclear factor-kB ligand (RANKL) and monocyte-colony stimulating factor (M-CSF) in the presence/ absence of interleukin-1β (IL1β), tumor necrosing factor-α (TNFα) and IL17 (10ng/ml). Osteoclasts differentiation was assessed by expression of tartrate-resistant acid phosphatase (TRAP), carbonic anhydraseII (CA), matrix metalloproteinase-9 (MMP9), RANK and cathepsinK (CatK). Relative expression of CA, MMP9, RANK and CatK were increases in TNFα treated cells (p<0.05) with CCLr. Cells from CCLr and MPL increased the expression of CA and RANK with IL1β (p<0.05) and TNFα (p<0.05) respectively. In the dogs with CCLr, MMP9 and RANK expression were increased (p<0.05), when compared with MPL derived samples. There was mild expression level of RANK in fresh SM from CCLr and MPL though catK expression increased in CCLr. The results of this study are expected to be emphasis the important of investigating of biomarkers and biotherapies targeting osteoclasts precursors those are presenting in vary number in CCLr and MPL to minimize joint inflammation and structural damage.
1. Lai Lai San (Division of Bioresources, CZC, D2)
"MIRU-VNTR Typing of Beijing multidrug resistant tuberculosis strains from Myanmar"
Introduction
Myanmar is one of the countries with the highest burden of tuberculosis (TB) as well as multidrug-resistant tuberculosis (MDR-TB). The previous studies on genotping in Myanmar pointed out that the Beijing strain has a significant association with MDR-TB. The active transmission of MDR Mycobacterium tuberculosis (MTB) strains is the emerging problem, hence, it is necessary to identify characteristics that can explain why they keep spreading widely. A total of 212 Beijing MDR-MTB isolates collected from Myanmar was analyzed by variable number of tandem repeat(s) of mycobacterial interspersed repetitive units (MIRU-VNTR) typing. Fifteen loci of MIRU-VNTR ( QUB26, Mtub21, MIRU31, MIRU10, QUB11b, MIRU26, MIRU4, MIRU39, MIRU40, Mtub4, QUB4156, Mtub30, MIRU39, ETR A, MIRU16) were used and the results were analyzed by bionumeric software. Thirty-eight clusters in 212 isolates with the biggest cluster consisted of 8 isolates were found. VNTR set used in this study was useful for the discrimination of this genotype and effective for the determination of transmission pattern of MDR-TB. Beijing MDR-MTB was highly diverse and it can be inferred that the outbreak of MDR-TB is less likely to occur. MDR-MTB may be emerging in each patient individually. Therefore, we can stop spread of MDR-MTB with proper drug usage.
2. Tatsuya Deguchi (Lab. of Veterinary Surgery, D1)
"Analysis for radioresistance of cancer stem-like cells derived from canine tumors"
Introduction
CSCs have been defined as a small population of cancer cells that are resistant to radiotherapy and their survival after radiotherapy has been linked to recurrence and metastasis. Various studies have been performed to characterize human CSCs and in the canine existence of CSCs has been reported. However, it is uncertain on the level of radioresistance of these canine CSCs. My research is to identify and characterize canine CSCs and the level of radioresistant and how they relate to prognosis. In this study, CSCs of various canine cancer cell lines are identified by using sphere formation assay. The stemness of spheroid cells derived from canine cancer cells was evaluated by expression of stemness markers, tumorigenesis and investigated radioresistance. Spheroid cells from canine osteosarcoma cell line were identified. Spheroid cells showed expression of stemness markers CD133 gene. In future, spheroid cells will be investigated for expression of other stemness markers and radioresistance evaluated and compared with adherent-cultured cells. The expression of CD133 marker would suggest that spheroid cells possess stemness and provide a CSCs research tool to investigate CSCs of canine tumour cells.
3. Ochirkhuu Nyamsuren (Lab. of Infectious Disease, D3)
"Study on epidemiological and immunological research for intractable infectious diseases in livestock"
Introduction
Out of 339 samples collected from cattle in the Luzon island of the Philippines, 324 (95.5%), 154 (45.4%), 209 (61.6%), 140 (41.3%), and 2 (0.6%) were positive for Anaplasma marginale, Babesia bigemina, Babesia bovis, Theileria spp., and Trypanosoma evansi infections. Phylogenetic analyses showed that A. marginale 16S rRNA gene was same lineage across the world, most of B. bovis RAP-1 and B. bigemina AMA-1 genes detected were same and similar lineage from Asian isolates, and the MPSP genes of Theileria spp. belonged to the group of T. sergenti and T. buffeli. On the other hand, molecular epidemiological studies of several infectious diseases were done in Mongolian livestock, 3 (0.84%), 20 (3.86%), 11(8.7%) and 97 (23.2%) were positive for Mycobacterium avium subspecies paratuberculosis, Bovine leukemia virus, Bovine viral diarrhea viruses and Anaplasma marginale in cattle samples by ELISA, nested PCR and RT-PCR assays. In addition, Anaplasma ovis was detected in other ruminant species and infection rate was 3 (33.3%) of 100 yak, 189 (94%) of 201 sheep and 73 (36.5%) of 200 goat samples, respectively. The phylogenetic analysis of Bovine leukemia virus, Bovine viral diarrhea viruses Anaplasma marginale and Anaplasma ovis were performed and compared with other isolates from several countries.
4. Mizuho Suzuki (Lab. of Microbiology, D2)
"Exploring the mechanisms of interspecies transmission of H2 influenza viruses and vaccine preparation for a future pandemic"
Introduction
H2N2 influenza viruses caused a pandemic in 1957, but have not been detected from human population since 1968. H2 influenza viruses might be re-introduced into humans in the future because the H2 influenza viruses are still circulating in birds. Since most of human population are immunologically naive to H2 influenza viruses, it would cause a pandemic. Previous studies revealed that human pandemic influenza viruses originated from avian influenza viruses and emerged via pigs. The aim of this study is to evaluate pandemic potential of avian H2 influenza viruses based on their receptor specificity, and to prepare a vaccine for a future pandemic. During three-time passages of A/duck/Hong Kong/278/1978 (H2N9) in pigs, the recovered viruses did not show specificity to human-type receptors. The H2N9 influenza viruses will be passaged a few more time to clarify the pandemic potential of avian H2 influenza viruses. In order to prepare a vaccine, we analyzed avian, human and swine H2 influenza viruses genetically and antigenically. Antigenicity of H2 influenza viruses did not differ each other despite their genetic diversity. Based on this result, we will select vaccine candidate strain which growth effectively in chicken embryonated eggs and possess enough immunogenicity in mice.
1. Jinxi Wang (Lab. of Experimental Animal Science, D2)
"Studies of the Influence of Genetic Background on the Severity of Hirschsprung's Disease by Using Rat Models"
Introduction
The Hirschsprung's Disease (HSCR) is an inherited disease controlled by multiple genes with complicated genetic mechanism. Patients with this disease suffer various extent of constipation including complete intestinal obstruction. Up to now, although more than 11 genes have been identified as responsible for this disease, the vast majority (about 80%) of HSCR heritability is still hidden or missing. The Ednrb gene is one of the most important genes for the development of HSCR. By studying three different rat strains carrying the same Ednrb mutation that show a big difference in their symptom, we found 17 SNPs in the critical genomic region responsible for the different severity in mutated Ednrb gene-induced HSCR. To confirm our idea and identify the specific SNP/SNPs, we selected another two rat strains each possessing the same haplotype as rat strains showing mild and severe symptoms, respectively, and induce a mutation on their Ednrb gene by using GONAD (Genome-editing via Oviductal Nucleic Acids Delivery) method. Through this investigation, we expect to find out some hidden genetic mechanisms of the HSCR, which will help further studies of this disorder.
2. Md. Abdul Masum (Lab. of Anatomy, D1)
"Vascular associated stem cell niche and its pathological alternation in mouse kidney"
Introduction
In this study, the author firstly examined the alternation of blood vessel structures in the kidney of CKD model mice. BXSB/MpJ-Yaa mice clearly developed glomerulonephritis with tubulointerstitial lesions. B- and T-cells were infiltrated to both glomerulus and tubulointerstitium, and macrophages were diffusely found in peri-glomerular or tubulointerstitial regions. These cell infiltrations were severer at late stage compared to early stage, and significant increase was detected in B-cells (P < 0.01). IL-1F6+ injured renal tubules were more abundantly observed at late stage compared to early stage. No myofibroblasts were found in tubulointerstitial lesions at both stages. For vasculature, no significant difference was observed in the number of peri-tubular capillaries contacting with renal tubules between early and late stage. However, CD34+ peri-tubular capillary/unit area in the kidney was significantly decreased at late stage compared to early stage (P < 0.01). In examined parameters, the decreased CD34+ peri-tubular capillaries were significantly correlated with the increased B-cells (P < 0.001, ρ = -0.812). Genetic mutation in Y chromosome of BXSB/MpJ-Yaa mice causes autoimmune disease phenotypes, especially systemic increase of autoreactive B-cells. Our results indicated the negative correlation between B-cell infiltrations and decreased CD34+ capillaries with the progression of CKD. From the results of histoplanimetry, the author considered that altered microvascular environment by inflammation might affect CD34 expression in endothelial cells without the quantitative changes of capillaries.
3. Shirin Akter (Lab. of Parasitology, D3)
"Serological and molecular survey of Leishmania infection in stray dogs of visceral leishmaniasis endemic areas in Bangladesh"
Introduction
Transmission of VL caused by Leishmania Donovani is considered to be anthroponotic (human to human transmission) in the Indian subcontinent. L. infantum which is phylogenetically most related to L. donovani is found in the Mediterranean, Middle East, Asia, and South American countries, and responsible for zoonotic VL, with dogs as the main reservoir host. However, limited studies have been conducted to investigate the role of dogs in VL caused by L. donovani. In 2012, a total of 50 stray dogs in VL endemic areas of Bangladesh were screened for Leishmania infection. Anti-Leishmania antibodies were detected in 6 (12%) dog serum samples using rK39 immunochromatographic stick test (ICT). Five (10%) samples were positive for Leishmania DNA using ITS1-PCR. Leishmania species was identified as L. donovani by sequencing of the amplified products. In order to demonstrate the presence of Leishmania in dogs, another 30 dogs were captured in 2013 from the same sampling area. The microscopic examination of peripheral blood smears was negative for Leishmania infection in all dogs. The rK39 ICT detected one positive dog, which was sacrificed to collect lymph nodes, liver and spleen. Both microscopic analysis and PCR were negative for these tissue samples. Although our findings support the assumption that dogs might be a probable animal reservoir for L. donovani, further studies are required to understand the role of dogs in the transmission VL.
4. Chihiro Kanno (Lab. of Theriogenology, D2)
"Development of the procedure for simultaneous evaluation of multiple characteristics of bovine spermatozoa by flowcytometry"
Introduction
The objective of this study was to develop an objective evaluation procedure to estimate viability, acrosomal integrity, and mitochondrial membrane potential of bull spermatozoa simultaneously by flowcytometry. Firstly, we used frozen-thawed semen mixed with 0%, 25%, 50%, 75% and 100% dead spermatozoa. Semen was stained using three staining solutions: SYBR-14, propidium iodide (PI), and phycoerythrin-conjugated peanut agglutinin (PE-PNA), for the evaluation of spermatozoon viability and acrosomal integrity. Then, the characteristics evaluated by flowcytometry and by fluorescent microscopy were compared. In terms of the results, the characteristics of spermatozoa evaluated by two procedures were similar. Secondly, we attempted to evaluate viability, acrosomal integrity, and also mitochondrial membrane potential of spermatozoa by flowcytometry using conventional staining (SYBR-14, PI, and PE-PNA) combined with MitoTracker Deep Red (MTDR) staining. Then, the spermatozoon characteristics evaluated by flowcytometry using quadruple staining were compared with those of staining using SYBR-14, PI, and PE-PNA and staining using SYBR-14 and MTDR. From the obtained results, there were no significant differences in all characteristics of spermatozoa evaluated by quadruple staining and the other procedures. In conclusion, staining procedure using SYBR-14, PI, PE-PNA, and MTDR for flowcytometry can evaluate viability, acrosomal integrity, and mitochondrial membrane potential of bovine spermatozoa simultaneously.
1. May June Thu (Unit of Risk Analysis and Management, CZC, D2)
"Genetic characterization of Rickettsia spp. circulating in Japan"
Introduction
Rickettsiae are obligate intracellular gram-negative bacteria that cause rickettsioses in humans. They are divided into two groups; the spotted fever group (SFG) transmitted by ticks and the typhus group (TG). In Japan, approximately 40 cases of Japanese spotted fever (JSF), caused by Rickettsia japonica, are reported annually. The previous studies reported only partial sequences of rickettsiae circulating in Japan. Therefore, isolation of diverse Rickettsia including non-JSF Rickettsia, followed by genetic characterization at the whole genome level, is required to evaluate the potential risk of the emergence of non-JSF rickettsiosis. From 2013 to 2015, a total of 2,175 ticks, including 17 different tick species, were collected from 13 different prefectures in Japan. DNA samples were examined for Rickettsia infection by real-time PCR. Positive samples were used for genetic characterization by conventional PCR and sequencing. Out of 2,175 ticks, 404 samples (18.6%) were infected with Rickettsia spp. Sequencing analysis of the amplified PCR products indicated the presence of 19 different genotypes. Tick homogenates of Rickettsia-positive samples were inoculated into ISE6 (tick) and C6/36 (mosquito) cell lines. Six different genotypes of Rickettsia spp. were isolated from ISE 6, while one genotype was isolated from C6/36.
2. Ai Dantsuka (Lab. of Anatomy, D1)
"Elucidation of biological characteristics determined by genomic diversity using MRL/MpJ as a mouse model"
Introduction
The mammalian genome contains its own genetic information. Genomic diversity is crucial for the species-specific characteristics in organ morphology and function. Genomic mutations also contribute to their developments, however, they often cause diseases in both humans and animals. Experimental animals, especially inbred mice, are valuable to clarify the relationship between genetic factors and phenotypes because of their abundant biological information.
Inbred MRL/MpJ mice show several unique phenotypes in urogenital organs or immune system caused by the genetic factors in MRL/MpJ-type genome. To discover novel genetic factors making MRL/MpJ unique, I established genetically modified MRL/MpJ, carrying transgenic oncogene or a mutation of muscular dystrophy. Then their phenotypes are being compared to those in other inbred strains.
In this study, I introduce a transgenic MRL/MpJ-Tg(SV40-Ahr)1Poe, expressing the constitutively active aryl hydrocarbon receptor (CA-Ahr) under the control of SV40 promoter. Although original-type (FVB/N) CA-Ahr mice develop stomach tumors in the glandular part, the macroscopic severity of tumors in CA-Ahr mice seemed to be milder in MRL/MpJ-background than in original-type. Therefore, MRL-MpJ mice could be a useful model to elucidate genomic diversity, in particular in the genomic factor-dependent differences of tumorgenesis.
3. Nao Eguchi (Div. of Global Epidemiology, CZC, D2)
"The Mechanism of Inhibition of Influenza Virus Infection by M2 specific Antibody"
Introduction
The antigenic variability of Influenza A virus (IFV) hemagglutinin (HA), which is the principal target of neutralizing antibodies, is a major problem for influenza vaccine manufacturing. In contrast, the M2 protein, one of the envelope proteins of IFVs, is known to have highly conserved amino acid sequences regardless of the HA antigenicity, and expected as a potential target for universal influenza vaccines. It was shown that a monoclonal antibody rM2ss23 recognizing the M2 extracellular domain (M2e) did not neutralize infectivity of an influenza A virus strain, A/Aichi/2/68 (H3N2) (Aichi), but reduced the number and size of plaques of this strain when infected cells were cultured in the presence of the antibody. However, interestingly, rM2ss23 did not show any antiviral effects against another strain, A/Puerto Rico/8/34 (H1N1) (PR8), despite its ability to similarly bind to M2e of PR8. A reverse genetics approach revealed that rM2ss23 reduce the plaque formation of the reassortant viruses containing Aichi HA and M segments but not that of the virus containing M segment alone. For future research, we are planning to elucidate the antiviral mechanism of rM2ss23 in more detail.
4. Kochi Toyomane (Div. of Infection and Immunity, CZC, D2)
"Investigation of the molecular mechanism of AtxA-mediated virulence regulation on Bacillus anthracis"
Introduction
Anthrax is caused by Bacillus anthracis, a Gram-positive, spore-forming bacterium. The pathogenicity of this bacterium depends on two plasmids encoding the most important virulence factors. One encodes anthrax toxin composed of protective antigen, edema factor and lethal factor, while the other encodes capsule synthases. Expression of these factors is coordinately induced at transcription level. This regulation is mediated by a transcription factor of B. anthracis, AtxA. Since it is reported that deletion of atxA leads to low production of virulence factors and attenuation of B. anthracis, AtxA is thought to be a key factor for pathogenicity of anthrax. In spite of its important role in B. anthracis infection, the molecular mechanism by which AtxA controls the expression of virulence factors is not elucidated. To identify and characterize the factors regulated by AtxA directly, we constructed a reporter system. Using the system, we evaluated the interaction between AtxA and a promoter region that is predicted to be binding region of AtxA. As a result, the direct interaction between AtxA and the promoter region was not observed. It is suggested that AtxA controls the transcription of the target genes by unknown mechanisms rather than direct interaction with the promoter regions.
1. Khoirun Nisa (Lab. of Internal Medicine, D2)
"The Application of Contrast-Enhanced Ultrasonography (CEUS) of Duodenum to Diagnose Chronic Enteropathy in Dogs: Repeatability study"
Introduction
Contrast-enhanced ultrasonography (CEUS) with microbubble as contrast agent can detect pathological changes in intestine by qualitative and quantitative assessment of its perfusion. It may provide a non-invasive modality without radiation that not only support in finding definitive diagnosis but also enable disease monitoring of canine chronic enteropathy (CE). Repeatability of duodenal CEUS is necessary for technique validation and determining reliable measurements to detect changes related to intestinal disorders. Five healthy beagles (age 1-4 year, body weight 9-12 kg) were enrolled for this study. Duodenum was imaged following intravenous injection of Sonazoid® (0.01 ml/kg). Region of interest (ROI) were drawn in duodenal mucosa to create time-intensity curve (TIC). Perfusion parameters including time [wash-in 30% (WI-30), wash-in 70% (WI-70), time-to-peak (TTP), wash-out 70% (WO-70), wash-out 30% (WO-30)] and intensity parameters [peak intensity (PI), area-under-curve (AUC), peak-phase-rate (PPR)] were derived from TIC. CEUS was performed three times within one day (4-hour-interval) and in two different days (1-week-interval) to evaluate intraday and interday variability, respectively. Both time and intensity parameters except WO-30 were relatively stable with intraday and interday coefficient of variation (CV) for most parameters ≤20% (range, 8.30% to 20.31%). Duodenal CEUS could be reliable to evaluate changes related to CE in dogs.
2. Jeewan Thapa (Div. of Bioresources, CZC, D3)
"Molecular characterization of Mycobacterium orygis isolates from animals of South Asia"
Introduction
Mycobacterium orygis, a new member of Mycobacterium tuberculosis complex (MTC) species cause tuberculosis (TB) in humans and animals. We have isolated this bacterium from wild animals in Nepal and captured monkeys and dairy cattle in Bangladesh. MTC isolates from a spotted deer and a blue bull in Nepal and from a dairy cattle herd and captured rhesus monkeys in Bangladesh were included. Most of these animals had TB suspected lung lesions, these lesions were later cultured in LJ media. Spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR), region of difference analysis and SNP detection were performed to ascertain species and molecular epidemiology of the isolates. All the isolates had a typical spoligotype, SIT587 in SpolDB4 database, corresponding to M. orygis and in an agreement with many other reported isolates. RD analysis (cfp32: +, RD9 - and RD12-) and SNP detection of gyrB, mmpL6, TbD1, PPE55 and Rv2042c confirmed the isolates to be M. orygis. The MIRU-VNTR analysis of these isolates grouped them into four MIRU-VNTR types. Previous studies have reported M. orygis from humans in South Asia. These finding suggests its endemic prevalence in the sub-continent. Moreover, genetic diversity of these isolates indicates a long history of prevalence in South Asia. We hope these findings can be helpful to understand the situation of tuberculosis in the region.
3. Eugene Chisela Bwalya (Lab. of Veterinary Surgery, D3)
"Pentosan polysulphate downregulates iNOS, c-JUN and HIF-1α expression in IL-1β- stimulated canine articular chondrocytes"
Introduction
Stimulation with 1, 5, 10 and 20 ng/mL rhIL-1 for 8 hours significantly upregulated (p < 0.05) iNOS, C-JUN, HIF-1α and HIF-2α mRNA expression. Pre-incubation of chondrocytes with 1, 5, 15 and 40 μg/mL PPS before rhIL-1β-stimulation resulted in a dose-dependent downregulation of iNOS, c-JUN, and HIF-1α relative to the positive control. c-JUN and HIF-1α were significantly downregulated (p < 0.05) at all PPS concentrations. Although both HIF-1α and -2α were upregulated by rhIL-1β and significantly associated (p < 0.001) with iNOS expression (R2 = 0.61, correlation coefficient = 0.8 for HIF-1α versus R2 = 0.61, correlation coefficient = 0.61 for HIF-2α), HIF-1α was significantly downregulated at all PPS concentrations suggesting a protective role of PPS from HIF-1α catabolic effects. HIF-1α (and not HIF-2α) may be involved in the induction of iNOS and promotion of cartilage matrix degradation. HIF-2α mRNA remained upregulated at almost all PPS concentrations suggesting a chondroprotective role. The expression of iNOS mRNA in rhIL-1-untreated chondrocytes suggest post-transcriptional regulation. The study has for the first time demonstrated that PPS can ameliorate IL-1β driven OA via effective inhibition of iNOS, c-JUN and HIF-1α mRNA.
4. Heidi Lynn Tessmer (Div. of Bioinformatics, CZC, D3)
"Computational Analyses of Host and Viral Genomes"
Introduction
As our ability to obtain genomic data increases, so does our ability to conduct in-depth analysis of diseases, uncovering previously unknown patterns of evolution and spread. In the future, our ability to effectively combat zoonotic infectious diseases, such as influenza and Ebola virus disease, will require a thorough understanding of the evolutionary mechanisms of the viruses. These mechanisms can most efficiently be discovered through computational analysis of genomes from the nucleotide and protein levels up. This research focuses on rudimentary analysis of single and dinucleotide content in numerous viral and host species. Previously identified trends in C+G content, as well as dinucleotide composition in these viruses, were corroborated and new viruses were analyzed, specifically the 2009 pandemic influenza virus and the Ebola virus from 1976 to the current epidemic. Our findings show similar patterns between human and avian H3N2 viruses, as well as between human influenza and the Ebola virus. Further research is needed to clarify whether these similarities are characteristic of all viruses or specific to the ones which we have chosen.
1. Nan Aye Thida Oo (Div. of Bioresources, CZC, D1)
"Detection of pncA gene mutations in multidrug-resistant Mycobacterium tuberculosis isolates from Myanmar"
Introduction
Pyrazinamide (PZA) is a first line anti-tuberculosis drug and used for both short course and multi-drug resistant tuberculosis (MDR-TB) treatment regimens. It is a pro-drug and activated by pyrazinamidase which is encoded by pncA. Numerous studies reported that mutations of pncA were responsible for PZA resistance by losing or reducing PZase activity. The aim of present study was to detect the frequencies and patterns of pncA mutation among MDR-TB strains isolated in Myanmar. Clinical MDR-TB isolates were collected and PCR amplification and DNA sequencing of pncA were carried out to detect mutations. Of 369 isolates, 230 samples were processed for DNA sequencing. Mutations in pncA were identified in 96 (41.7%) isolates, while 134 (58.3%) samples had wild type sequences. A total of 69 different types of mutation were dispersed on the pncA and thirty-four of which were found to be novel mutations. This study showed that there were new and diverse mutations on pncA and further studies of correlation between mutations in pncA and alteration of pyrazinamidase activity are needed. Mutation patterns of pncA can be applied for the future development of rapid genotypic PZA susceptibility assays which can lead to great achievement in tuberculosis treatment and control strategies.
2. Yusuke Izumi (Lab. of Advanced Veterinary Medicine, D2)
"Isolation and characterization of tumor endothelial cells"
Introduction
Tumor angiogenesis is the growth of new blood vessels from the existing vasculature toward tumor tissues. Tumor vasculature supplies nutrients and oxygen. It is necessary to tumor growth and metastasis.
Anti-angiogenic therapies function by inhibiting tumor angiogenesis. It causes tumor death indirectly. Several anti-angiogenic therapies have been developed, including bevacizumab, toceranib, and metronomic chemotherapy. Conventional anti-angiogenic drugs target not only tumor endothelial cells (TECs), but also normal endothelial cells (NECs), which may cause adverse effects.
Recently, a few reports indicated that the morphology and genetic expression of human and mouse TECs might differ from NECs. However, there is quite limited information about canine TECs Understanding the characteristics of TECs is essential to the establishment of a new generation of anti-angiogenic therapies with fewer adverse effects. The present study involved the comparison of the characteristics of canine TECs.
In my preliminary experiment, I revealed that canine TECs have different characteristic compared to NECs, such as PD-L1 and CXCR7 expression in the limited histophathological specimens.
In my future study, I would like to do immunohistochemical study using CD105, kind of TEC marker, of various canine tumor tissues and isolate canine TECs using magnetic cell sorting system.
3. Paulina Duhita Anindita (Div. of Molecular Pathobiology, CZC, D2)
"Establishment of compound screening assays for identification of novel antivirals against rabies virus"
Introduction
Rabies is an invariably fatal neurological disease predominantly in rural areas of Asian and African countries caused by rabies virus (RV). Currently available post-exposure prophylaxis procedure and antiviral therapies are not effective for RV-infected individuals after onset of symptoms. Thus, it is urgent to develop new antiviral therapies as an approach to treat exposed and symptomatic individuals. Novel antiviral agents can be identified by screening of compound libraries. As it is known that RV infection does not generate easily discernible cytopathic effects, cell viability assays cannot be employed for such screening of small compounds. To overcome this issue, recombinant RVs (rRVs) encoding NanoLuc luciferase (NanoLuc) inserted in the intergenic region of the RV genome. We determined the growth kinetics of rRVs and demonstrated no decreases in NanoLuc expression over ten passages of virus. We further examined the usefulness of the rRVs for screening of antiviral compounds by treating rRV-inoculated cells with ribavirin, a nucleoside analogue which has a known anti-RV activity in vitro. We demonstrated that NanoLuc signals were decreased in a dose-dependent manner of ribavirin. These results showed that rRVs encoding NanoLuc are a useful tool for screening of antiviral compounds against RV.
4. Yinghua Yang (Lab. of Theriogenology, D2)
"The effect of bone morphogenetic protein 4 (BMP4) on in vitro growth and subsequent development of oocytes derived from bovine early antral follicles"
Introduction
Oocytes in small follicles are less competent than those in large follicles are. The inferior may be caused partly by suboptimal in vitro growth (IVG) culture system in which role of theca cells is absent. Theca cells are considered as an indispensable part to a follicle and one of their major products, BMP4, is an important regulatory factor during follicle development. This study was carried out to test the effects of BMP4 on oocyte IVG and subsequent development. Oocytes from small follicles were treated with three doses (0, 10 and 50 ng/ml) of BMP4 during IVG culture. Throughout IVG, progesterone production from granulosa cell (GC) was suppressed in BMP4 treated groups, consistent with delayed luteinization. After IVG, GC number and oocyte viability were significantly decreased in 50 ng/ml BMP-4 treated group compared to non-treatment group. For subsequent development of IVG oocyte, BMP-4 treated group did not bring improvement in nuclear maturation after in vitro maturation and fertilizability after in vitro fertilization while embryonic development was compromised. In conclusion, BMP4 addition during IVG culture showed the effect of anti-luteinization associated with decreased progesterone level; however, impaired GC proliferation, oocyte survivability and subsequent embryonic development.
1. Md. Atiqul Islam (Lab. of Experimental Animal Science, D1)
"Analysis for resistance/susceptible genes to Echinococcus multilocularis infection in the mouse"
Introduction
Establishment of congenic lines from B6 and D2 mice using microsettelite PCR could reveal significant resistance/susceptible genetic locus to E. multilocularis infection in the mouse. We already developed 7 congenic and subcongenic lines using microsettelite markers that include chromosomal regions involving the genetic locus responsible for cyst establishment and protoscolex development. For cyst establishment B.D-Emcys1 and D.B-Emcys1 congenic lines and for protoscolex development B.D-Empsc1, D.B-Empsc1, B.D-Empsc1sub1, B.D-Empsc1sub2, and D.B-Empsc1sub1 were established. We challenged these mice with E. multilocularis infection by oral administration of eggs of E. multilocularis. After infection experiments, we found that there was no significant effects of D6Mit188-D6Mit15 locus to cyst establishment. However, we found interesting results in subcongenic line B.D-Empsc1sub1 where 3 mice developed protoscolex among 7 mice. B.D-Empscsub1 subcongenic mice contains D1Mit191-D1Mit14 locus from D2 mice that might possess susceptible gene for protoscolex development. As there were 4 mice in this same line that did not develop protoscolex, so it is required to re-confirm the substituted genomic region. In future identification of the responsible gene and furthermore validation of responsible gene by generating knock-in mice with CRISPR/Cas gene-editing method would be necessary. Our research outcomes would lead novel treatment strategies to control Echinococcosis.
2. Naoya Maekawa (Lab. of Infectious Disease, D2)
"Development of a novel immunotherapy against dog tumors by targeting immunoinhibitory molecules"
Introduction
An immunoinhibitory receptor, PD-1, is expressed on tumor antigen-specific T cells, and suppresses the effector functions of these cells when it binds its ligand, PD-L1. PD-L1 is expressed on many types of tumors and thus plays an important role in the immune evasion of tumor cells. In the field of medical science, therapeutic effect of anti-PD-1 antibody, which inhibits the interaction of PD-1 with PD-L1, has been well characterized and this antibody is expected to be a new treatment for several types of malignancies including advanced melanoma. The purpose of this study is to apply this strategy to dog malignancies.
Immunohistochemical and flow cytometric analyses revealed that dog melanoma, mastocytoma, renal cell carcinoma and other types of tumors express PD-L1. In addition, the expression levels of PD-1 on tumor-infiltrating lymphocytes (TILs) were higher than those of healthy controls. Moreover, dog TILs treated with anti-PD-L1 antibody had improved abilities to produce IFN-gamma, suggesting the potential of this antibody as a novel drug against dog tumors.
Further analysis is required to confirm the therapeutic effect of our antibody on dog patients. After the in vitro analysis, we are planning to perform a clinical trial by using this antibody.
3. Jednipit Borthong (Div. of Bioinformatics, CZC, D2)
"Analysis of bacterial population in water samples collected from the Phadungkrukaseam Canal, Bangkok, Thailand using NGS"
Introduction
Phadungkrukaseam Canal is located in Bangkok, Thailand, is abundant with aquatic plants and animals, and is closely associated with human life. This canal also receives wastewater from household areas and several markets: fresh food, animals, and plants. Thus, this canal may contain a difference of bacterial population. However, this data is still remained and required. In this study, a total of 5 samples (A - E) were analyzed bacterial population based on the nucleotide sequences at the highly variable V3 - V4 region of 16S rRNA using Illumina Miseq Platform. Furthermore, Shannon's index and Simpson's index were used to determine genus richness while SØrensen-Disc coefficient and Kullback-Leibler (KL) divergence were used to compare the similarity of bacterial population. The results revealed that a total of 238 genera (belonging 28 phyla) were found, 27 genera are associated with infectious disease in human. Additionally, alpha diversity indicated that the point C was highly divergent than the other points. SØrensen-Disc coefficient displayed approximately 60 - 70% similarity of bacterial genera detected while KL divergence obviously certified significant difference between bacterial populations. Our results verified that this canal harbors different population of bacteria depending the points and contains some bacteria related with infectious diseases.
4. Minato Hirano (Lab. of Public Health, D2)
"Study on genomic transport-mechanism of the neuronal infection of flaviviruses"
Introduction
Neurological diseases caused by encephalitic flaviviruses are severe and are associated with high levels of mortality. However, detailed mechanisms of the viral replication and pathogenesis in the brain remain poorly understood. Previously, we reported that the replication of tick-borne encephalitis virus (TBEV) altered membrane structure in dendrites of mouse primary neuronal culture, which might be involvegd in the neuro-pathogenicity of TBEV. In the current study, I analyzed detailed mechanisms of the transport of the viral genome to the neurites. Transfection of the DNA-based TBEV replicon caused the viral antigen and genomic RNA accumulations in neurites of the PC12 cells. The viral genomic RNA was still observed in the cells transfected with a replicon not expressing the viral proteins. As well as in the cells transfected with a replicon in which most of the coding sequence was deleted. These results showed that the viral genomic RNA was directly transported to the neurites, and the transport was independent of the viral proteins. The RNA sequence of the 5'-UTR and/or 3'-UTR were required for the transport to the neurites. I will further examine the viral and host factors involved in the transport to reveal molecular mechanisms of TBEV infection in neuronal cells.
1. Masahiro Sato (Div. of Global Epidemiology, CZC, D1)
"Studies on the biological property of bat-derived influenza viruses"
Introduction
Influenza A virus (IAV) is a member of the family Orthomyxoviridae and have 8 segmented single-stranded negative-sense RNA genomes. IAV is one of the most important zoonotic pathogens. Recently, IAV genomes of novel subtypes (i.e., H17N10 and H18N11) were detected from frugivorous bats in Guatemala and Peru. However, since infectious virus strains have never been isolated, the biological properties of these bat-derived IAVs are largely unknown. In this study, we aim to elucidate receptor molecules required for cellular entry of bat-derived IAVs, and also attempt to obtain information on the host range and zoonotic potential of the virus. First, we will try to rescue a reassortant virus which has HA and NA genes of bat-derived IAV and the other genes of a well-characterized IAV strain (PR8) and use the virus for screening receptor molecules. We will further attempt to generate bat-derived IAVs with a complete set of the 8 gene segments to infect experimental animals. Second, receptor molecules of bat-derived IAVs will be identified by an expression cloning method using cDNA library from IndFSPT1 cells which are potentially susceptible to bat-derived IAVs. Third, we will generate monoclonal antibodies against bat-derived IAVs and identify neutralizing epitopes on HAs.
2. Yuri Sakai (Lab. of Radiation Biology, D2)
"The response of cancer-associated fibroblasts to ionizing radiation and its effect on cancer pathophysiology"
Introduction
To improve the efficacy of cancer radiotherapy, the effect of radiation on cancer cells in tumor has been studied for a long time. On the other hand, the growing evidence suggests that cancer prognosis after treatment is greatly influenced by tumor stroma. Cancer-associated fibroblasts (CAFs) are major components of tumor stroma and secrete various soluble factors, resulting in malignant progression and metastasis. However, there is little information about how CAFs respond to ionizing radiation. Therefore, the aim of this study is to reveal the effect of radiation on the secretory phenotype of CAFs and its effect on cancer pathophysiology. In this study, following studies are planned: (1) production of experimental CAFs (eCAFs), (2) analyses of the soluble factor and gene expression of irradiated eCAFs, (3) analyses of the effect of irradiated eCAFs on cancer cell phenotypes, and (4) analyses of the effect of irradiated eCAFs on cancer pathophysiology. For these experiments, cell cultures and materials for these studies were prepared and some preliminary experiments were performed. The tumorigenicity of BALB/c mouse-derived mammary carcinoma cell line EMT6 was confirmed by inoculating them into mammary fat pads of BALB/c mice, and the metastasis to other organs was investigated. As a result, they formed multiple metastatic lesions in the lung.
3. Chihiro Ishii (Lab. of Toxicology, D2)
"Lead (Pb) poisoning is still a serious problem in raptors from Japan"
Introduction
Lead (Pb) poisoning occurs widely among raptors. Raptors ingest Pb rifle bullet or shot fragments when they consume mammals or birds that were hunted with Pb ammunition. In Hokkaido, the northern island of Japan, the use of Pb rifle bullets for deer hunting has been prohibited since 2001. Incidentally, other areas of Japan, such as Honshu or Shikoku have few regulations and the current studies of Pb poisoning are little clarified. In this study, we measured the Pb concentrations and Pb stable isotope ratios (Pb-IRs) to identify the source of poisoning in 16 species of 174 raptors from Japan. Pb distribution in bones and bone marrows was also measured. This study clarified that accumulation of one third sea eagle which died in Hokkaido exceeded the level of Pb poisoning after the regulation. In Honshu and Shikoku, 5 eagles include endangered species accumulated high Pb concentration, indicating that they were exposed to Pb. Pb-IRs indicate that both of Pb bullets and shot have the risk of causing Pb poisoning in all parts of Japan. Furthermore, there is a possibility that analysis of bones and bone marrows would be effective to reveal Pb exposure and periods of after exposure.
4. Yogendra Shah (Div. of Bioresources, CZC, D2)
"Diversity of MIRU-VNTR among Mycobacterium tuberculosis Central Asian strain family isolates from Nepalese patients"
Introduction
Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) poses major challenges and public health problems in Nepal. Central Asian Strain (CAS) family has been reported as one of the most predominant genotypes of MTB in South Asian countries including Nepal. The main objective of this study was to understand molecular epidemiological feature of MTB CAS Delhi strain circulating in Nepalese patients and to assess its transmission dynamics. A total of 145 MTB CAS isolates from Nepalese patients were analyzed by spoligotyping and 24 loci MIRU-VNTR. Using HGDI, three loci (QUB26, MIRU10, VNTR 424) were found to be "highly discriminatory" (DI>0.6), eleven loci (MIRU26, MIRU31, MIRU40, VNTR1955, VNTR 3960, ETR-A, VNTR 2401, QUB 4156, MIRU39, ETR-F and QUB 11a) were "moderately discriminatory" (DI:0.3-0.6) and ten loci (MIRU16, ETR-C, QUB11b, MIRU4, MIRU2, MIRU20, MIRU23, MIRU 24, MIRU27 and ETR-B) were poorly discriminatory (DI<0.3). By combining both MIRU-VNTR and Spoligotyping comprised eighteen clusters having the 47 isolates of identical pattern where as clustering rate was found to be 20%. The proposed 15 loci MIRU-VNTR typing scheme is well suited to assess the MTB population structure and diversity, trace back the transmission dynamics and epidemiological link among MTB CAS family strain within Nepal and surrounding countries.
1. Woongchul Shin (Lab. of Biochemistry, DC1)
"Effects of dietary component on the activation, proliferation, and recruitment of thermogenic cells"
Introduction
There are two kinds of fat tissue in mammals, brown adipose tissue (BAT) for heat generation and white adipose tissue(WAT) for energy storage. BAT exclusively expresses uncoupling protein 1 (UCP1), which is responsible for thermogenesis. Interestingly, in response to sympathetic nerve stimulation, WAT changes its metabolic phenotype to BAT-like one (WAT browning); UCP1-expressing brown-like adipocytes, termed as beige adipocyte are induced within the tissue. In the previous study, we found that ginger feeding suppressed diet-induced obesity in mice and that it activate brown adipocyte and also induced beige adipocyte. However, actual contribution of BAT activation and WAT browning to the anti-obesity effect by ginger is not clear. Thus, our purpose of study is to evaluate how brown and beige adipocytes contribute to the anti-obesity effects of ginger by using UCP1-KO mice. Also, although recent studies revealed that beige adipocyte arises from a progenitor cell different from brown adipocyte, the cellular origin of the progenitor is unknown. WAT is shown to contain mesenchymal stem cells, almost identical to those from bone marrow (BM). Also, it has reported that BM-derived stem cells migrate to WAT following to some pharmacological stimulation. Thus, we will examine whether BM-derived residential stem cells differentiate into beige adipocytes.
2. Teppei Ikeda (Lab. of Anatomy, D2)
"Studies on mechanism of skeletal muscle remodeling - Analysis of the molecules contributing to myoblast differentiation and growth -"
Introduction
All animals repeat disruption and regeneration of myofibers, that is, skeletal muscle remodeling. Aging or several diseases could cause disorder of skeletal muscle remodeling, which leads to breakdown and atrophy of skeletal muscle, resulting in astasia. These pathological progressions can be serious problems in livestock animals and elder companion animals since they get in difficulty in feeding, which leads to euthanasia and continuing care. Muscle repair is a multistep process that includes myofiber degeneration and regeneration. After damage to the myofibers, satellite cells activate, proliferate, and give rise to a population of transient-amplifying myogenic cells, called myoblasts, which express myogenic regulatory factors (MRFs) such as myogenic differentiation 1 and myogenic factor 5. Myoblasts subsequently express another MRF called myogenin, commit to terminal differentiation, and fuse to reconstruct their host myofibers or to generate new myofibers and repair the damaged tissue. Although much is known about the transcription factors and signaling pathways involved in the skeletal muscle remodeling, the detailed process is still incompletely understood. In this study, we widely investigated the morphological and pathological features of skeletal muscles in C57BL/10ScSn-Dmdmdx (mdx) mice, known as a representative model for Duchenne muscular dystrophy in humans.
3. Asami Nishimori (Lab. of Infectious Disease, D2)
"Large-scale production of the monoclonal antibody against bovine immunoinhibitory molecules for clinical application in cattle"
Introduction
In previous study, we revealed one of the disease developing mechanisms in bovine leukemia virus (BLV) infection. Programmed death-1 (PD-1) is a receptor on T cells, and the interaction of PD-1 and its ligand (programmed death-ligand 1: PD-L1) inhibits T-cell activation. Our previous reports showed that the both of PD-1 and PD-L1 expression were upregulated in line with disease progression of BLV infection, and moreover, the blockade of the PD-1/PD-L1 interaction by using antibodies increased IFN-γ production in vitro. These findings suggest that the PD-1/PD-L1 blockade could be clinically applied as a novel therapeutic method for controlling BLV infection. The next stage of our research is to evaluate the in vivo function of the blocking antibody. However, there are several problems to apply this antibody in cattle; 1) because the blocking antibody is delivered from rats, it might induce severe inflammation if frequently administered; 2) a large amount of antibody is required for clinical trial in cattle. Thus, we are planning to develop rat-bovine chimeric antibody and to establish cell clones for large-scale antibody production. Now, cell groups we established were producing 21.12mg/L of the chimeric antibody, and further steps to obtain more superior cell clones is ongoing.
4. Yuji Wada (Div. of Molecular Pathobiology, CZC, D2)
"Development of novel chemical therapy for Chikungunya fever"
Introduction
Chikungunya fever (CHIKF) is caused by Chikungunya virus (CHIKV) belonging to genus Alphavirus, family of Togaviridae. The main symptom of CHIKF is arthritis which often persists several months to over year, and sometime fatal for children, elderly and the persons who have underlying condition. CHIKF has now widely spread and caused severe damage for health and economic in the world because of no effective treatments and approval drugs. In order to control CHIKF, it is necessary to develop therapeutic strategies. First, we conducted the screening of chemical compounds library to find candidate compounds which inhibit CHIKV infection. As a result, we found the some candidate compounds. Additionally, we confirmed one of the candidate compounds inhibits CHIKV replication significantly. To unveil the CHIKV inhibitory mechanism of these compounds, we performed selective amplification of CHIKVs (res-CHIKV) whish are resistant to the inhibitory effect of candidate compounds. Subsequently, we performed full genome sequence analysis to find characteristic amino acid change in res-CHIKVs. As a result, we found three or four characteristic amino acid change from two clones of res-CHIKV. Now, we assess the relation between each mutation and resistant ability against candidate compounds by reverse engineering.
1. Reiko Akamatsu (Div. of Infection and Immunity, CZC, DC2)
"Multilocus sequence typing of Bacillus cereus nosocomial isolates"
Introduction
Bacillus cereus is associated with food-borne illness with vomiting and diarrhea. It is known that several strains among B. cereus potentially cause severe extra-intestinal infections. Pathogenic B. cereus caused nosocomial outbreaks and were lethal to patients in Japan. Based on multilocus sequence typing (MLST) and phylogenetic analysis, pathogenic B. cereus, which were isolated in Gifu University hospital and Jichi Medical University (GTC strains), are closely related to B. anthracis than B. cereus standard strain. However it is difficult to discriminate pathogenic B. cereus strains from less virulent or non-pathogenic strains. In 2013, National Cancer Center (NCC) had an outbreak of B. cereus infection. Twenty-nine strains of B. cereus were isolated from human blood samples and hospital appurtenances. From the result of MLST, we found that the properties of 8 strains were similar to GTC strains. There is a possibility that NCC outbreak was caused by B. cereus strains which were similar in nature to GTC strains.
2. Chiho Kaneko (Div. of Risk Analysis and Management, CZC, D2)
"Studies for enhancement of strategies for the control of rabies in Zambia"
Introduction
Rabies still causes 55,000 deaths per annum, predominantly among children in Asia and Africa. The execution of control of dogs undertakes the success of control of rabies, yet the situations in the endemic countries are confronted with various challenges. This research aims to understand the current status of rabies and evaluate the current control measures against rabies in Zambia. It will be succeeded by the risk analysis of rabies in Zambia using relevant data. The comprehensive outcome will apply to establish the future strategy for the control of rabies. The field survey collecting canine blood samples was conducted to evaluate the immunization coverage against rabies in domestic dogs and to evaluate the efficacy of domestic produced rabies vaccine. For the collection of canine blood samples, the mass vaccination campaigns were held in the target local districts. 736 dogs were vaccinated and then 265 blood samples were collected in total. In Lusaka, the capital of Zambia, two-stage cluster sampling was carried out. 278 canine blood samples were collected through the programme. KAP (Knowledge, Attitude and Practice) survey for the dogs' owners was also conducted and 224 responses were obtained. Subsequent analysis is in progress.
3. Suppalak Kaewkwan (Lab. of Wildlife and Medicine, D2)
"Genetic status monitoring of wild Asiatic black bears in Thailand forests"
Introduction
The Ursidae family is the largest carnivore group that has many published of brown bears and American black bears from Europe and North America. On the other hand, the status of wild Asiatic black bears in Thailand is little known, especially the genetic variation of this species has never been evaluated. Thai researchers are interested in wild bear ecology, habitat, food variety, and bear activity patterns. My study plan is focusing on genetic variation of wild Asiatic black bear in the forest complexes that are continuous evergreen forest in northern, northeastern and western Thailand. Hair samples will obtain from a rub tree trapping method using a DNA hair sampling technique. A set of hair trapping contained 20 coiled wires on the tree (5 rows and 4 columns) and a camera trap was set up at the opposite side. Wire has 1 inch with 0.5 inch spike at the end. All designed traps will be distributed in 5 wildlife sanctuaries with 6x6 km. (36 sq.km.) grid cell size that referred minimal Asiatic bear home range. We will put at least 10 traps near watershed areas where bears are commonly found.
4. Manyando Simbotwe (Div.of Infection and Immunity, CZC, D2)
"Development of an enzyme-linked immunosorbent assay for the sero surveillance of anthrax in endemic areas"
Introduction
Bacillus anthracis, protective antigen (PA) is composed of four structural domains and is responsible for the shuttling of edema factor and lethal factor into the host cell during the pathogenesis of anthrax. Each of the four domains of PA plays a role in the host intoxication process. Current serological assays including ELISAs have been developed based on PA, to analyze immune responses against B. anthracis infection. However, one of their major limitations is their inability to distinguish vaccine-induced immunity from natural infection. In order to effectively carry out sero surveillance and evaluate the immunization status of cattle in anthrax-endemic areas, the development of an assay that will overcome the above mentioned limitation is necessary. In this study, to prepare antigens for the new ELISA, we cloned and expressed each of the four PA domains individually. The recombinant proteins PA-D1, PA-D2, PA-D3 and PA-D4 were expressed as GST fusion proteins in Escherichia coli. The results of the protein expression revealed that PA-D1 was expressed in the soluble fraction of the cell lysate. On the other hand, PA-D2, PA-D3 and PA-D4 had low levels of expression in the soluble fraction. Predominant expression of the proteins was in the pellet fraction as inclusion bodies.
5. Motofumi Suzuki (Lab. of Radiation Biology, DC3)
"MPS1 inhibition enhances chemosensitivity to etoposide and paclitaxel but not radiosensitivity in murine SCCVII cells"
Introduction
In eukaryotes, the spindle assembly checkpoint (SAC) is an mechanism to ensure the proper chromosome segregation. Recently, several studies have demonstrated that MPS1 plays an important role in SAC signaling. Furthermore, MPS1 inhibitors have been shown to exhibit potent cytocidal effect, implying the potential of MPS1 as a novel target of cancer treatment. However, it remains to be determined how MPS1 inhibition influences the cell-killing effect by ionizing radiation or anti-cancer drugs. Therefore, our aim in this study is to elucidate the relationship between MPS1 and cellular radio-/chemo-sensitivity. To evaluate whether MPS1 inhibition influences cellular radio/chemosensitivity, we analyzed cellular survival after exposure to cytotoxic treatments with or without MPS1 inhibitors by clonogenic survival assay. Whereas both MPS1 inhibitors sensitized SCCVII cells to anticancer drugs, they did not enhance cellular radiosensitivity. Furthermore, to evaluate the effect of MPS1 inhibitors on cell cycle progression, we assessed cell cycle distribution after cytotoxic treatments with or without MPS1 inhibitors by flow cytometry. We found that tetraploid cells were increased when cells were co-treated with MPS1 inhibitors and anticancer drugs, while MPS1 inhibition had little effect on cell cycle status after irradiation. This study revealed that MPS1 is a potential target to improve cancer chemotherapy.
1. Wallaya Phongphaew (Div. of Molecular Pathobiology, CZC, DC2)
"Role of the Endoplasmic Reticulum-associated protein degradation (ERAD) in West Nile virus infection"
Introduction
ERAD is protein quality control of cells. It is related to degradation of misfolded-proteins to prevent cell death caused by accumulation of abnormal proteins. The strategies can be divided into four major steps including misfolded-protein recognition, retranslocation the misfolded-proteins into cytosol, ubiquitination to target for proteasome and degradation by proteasome. However, ERAD is not only protein quality control, but it also plays some roles in infection of many viruses. Human immunodeficiency virus type 1 (HIV-1) and herpesviruses manipulated ERAD to evade from host immune response. While hepatitis c virus (HCV) was increased when ERAD or ERAD components were inhibited. HCV envelope protein was target for ERAD. Decrease in ERAD activity could retain HCV envelope protein in the infected cells and increase production titer of infectious particles. On the other hand, the present results indicate that inhibition of ERAD using chemical compound, Eeyarestatin 1 decrease WNV infection. This compound target SEC61 and Valosin containing protein, the components in ERAD machinery. However, the mechanism remains unknown and requires further study to elucidate. We have planned to use other ERAD inhibitor and ERAD gene silencing to confirm whether ERAD inhibition effect to WNV infection and reveal that which steps of WNV replication is involved in ERAD machinery.
2. Yuji Hirai (Lab. of Veterinary Hygiene, D2)
"Analyses of cell-attachment and growth of Listeria monocytogenes in intestinal models for identification of novel virulence-associated markers"
Introduction
Listeria monocytogenes (Lm) causes severe and fatal foodborne diseases, such as gastroenteritis, meningoencephalitis, and maternofetal infections. Lm is isolated from patients, food products, and the environment. There are several genetic analyses for estimating the virulence of Lm isolates, however, genetic analyses are still short of the estimation of pathogenicity of Lm isolates. Thus, further virulence-associated markers are required to estimate the pathogenicity of isolates. In Lm infections, Lm multiplies in the intestinal lumen, cross the intestinal barrier and disseminate to the mesenteric lymph nodes, spleen, and liver upon ingestion of contaminated food. Cytotoxicity of Lm to intestine cells may provide the information for the assessment to the pathogenicity of Lm. Thus, we attempted to evaluate interactions of Lm with intestinal cells on cell-attachment, intracellular growth, and cell-to-cell spread. In this year, we carried out biofilm forming assay, cell-attachment assay, and plaque forming assay. We tentatively selected 20 representative isolates, including 9 clinical isolates and 11 food isolates, from 171 isolates based on the results of biofilm assay, and used them for following analyses. No difference was observed in results of clinical or food isolates in cell attachment assay and plaque forming assay. Hierarchical cluster analysis revealed that the 20 representative isolates were successfully classified into three clusters based on results of three in vitro assays. In the future, fine cluster analysis will be carried out using a number of Lm isolates and additional parameters, including molecular epidemiological characters. Pathogenicity and whole genome sequences of the representative isolates of each cluster will be analyzed for the identification of novel virulence markers.
3. Lesa Angela Thompson (Lab. of Toxicology, D2)
"Chicken - source of nutrition or toxin?"
Introduction
WHO estimated that in 2013 there were 198 million cases of malaria, with 584,000 deaths. 90% of these deaths occur in Africa. The disease is caused by species of Plasmodium parasite and spread by Anopheles mosquitoes. A common method of control is to use pesticides such as dichloro-diphenyl-trichloroethane (DDT) to reduce numbers of the vector. However, this chemical has toxic effects in many species - problems such as population declines in wild avian species resulted in the banning of DDT in many countries and its use is now strictly regulated. Although thought to be relatively safe for humans, reports have shown bioaccumulation to occur and suggested exposure may result in neurotoxic, carcinogenic, immunotoxic and reproductive effects. Muscle samples from free-ranging chickens (n=48) in the KwaZulu-Natal region of South Africa, where DDT is sprayed annually in homes were analysed for DDT content. The predominant DDT congener detected was the p,p'-DDE metabolite (mean, 10.09ug/g wet weight). The mean ΣDDT detected in free-ranging chicken muscle was 11.98ug/g wet weight. However, most chicken eaten in the area is intensively-reared broilers purchased locally. Analysis of such broilers (n=6) showed much lower levels of ΣDDTs (mean, 0.09ug/g wet weight).
4. Memi Muto (Lab. of Public Health, D2)
"Isolation and characterization of tick-borne encephalitis virus from Ixodes persulcatus in Mongolia in 2012"
Introduction
Tick-borne encephalitis virus (TBEV) is a zoonotic virus belonging to the genus Flavivirus, in the family Flaviviridae. The virus, which is endemic in Europe and northern parts of Asia, causes severe encephalitis. Tick-borne encephalitis (TBE) has been reported in Mongolia since the 1980s, but details about the biological characteristics of the endemic virus are lacking. In this study, 680 ticks (Ixodes persulcatus) were collected in Selenge aimag, northern Mongolia, in 2012. Nine Mongolian TBEV strains were isolated from tick homogenates. A sequence analysis of the envelope protein gene revealed that all isolates belonged to the Siberian subtype of TBEV. Two strains showed similar growth properties in cultured cells, but their virulence in mice differed. Whole genome sequencing revealed only thirteen amino acid differences between these Mongolian TBEV strains. Our results suggest that these naturally occurring amino acid mutations affected the pathogenicity of Mongolian TBEV. Our results may be an important platform for monitoring TBEV to evaluate the epidemiological risk in TBE endemic areas of Mongolia.
5. Tomoyuki Yamaguchi (Div. of Bioresources, CZC, DC3)
"The action of DNA gyrase and quinolone resistance"
Introduction
DNA gyrase is an essential enzyme for DNA replication and transcription of bacteria. It catalyzes the negative supercoiling of the bacterial circular DNA by cleaving the double strands, passing another part of DNA through the cleavage and resealing the DNA. Quinolone, a family of antibacterial drugs, interferes with the DNA-resealing of DNA gyrase and induce malfunction of DNA replication or transcription. Amino acid substitution in the quinolone binding site of DNA gyrase may confer quinolone resistance on bacteria. In Mycobacterium leprae that is the cause of leprosy, amino acid substitution at position 89, 91 and 95 on DNA gyrase subunit A can confer quinolone resistance. In our recent study, we conducted in vitro assays with moxifloxacin (MXF), sitafloxacin (SIT) and a newly developed quinolone to find out effective quinolones even against resistant strains. Their inhibitory activities on recombinant M. leprae DNA gyrases with four types of GyrA (wild type, Gly89Cys, Ala91Val and Asp95Gly) were examined and compared. We found SIT and the newly developed quinolone have much higher efficacy as DNA gyrase inhibitors than MXF.
1. Tapiwa Lundu (Lab. of Public Health, DC1)
"Studies on the ecology of Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV) and the interactions of viral proteins in infected cells"
Introduction
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonosis in China, Japan and South Korea, caused by SFTS virus (SFTSV), a novel phlebovirus in the family Bunyaviridae. The disease is characterized by fever, thrombocytopenia, coagulopathy and a case fatality rate of 2 to 15%. The SFTSV genome consists of L, M and S segments which encode viral RNA polymerase, glycoproteins (Gn and Gc), nucleoprotein (NP) and a nonstructural (NSs) protein, respectively. Haemaphysalis longicornis, Rhipicephalus microplus and Amblyomma testudinarium ticks are the vectors for SFTSV. Several mammals are potential reservoir hosts of SFTSV because anti-SFTSV antibodies have been detected in both wild and domestic animals. To determine the sero-prevalence of SFTSV antibodies in rodents, we used ELISA and IFA to test 1783 rodent sera collected between 1997 and 2012 in western Japan, Russia and Mexico. No antibodies were detected in rodent sera from the three countries. The result suggests a very low SFTSV prevalence in rodents during the survey period. It is suggested that NP and NSs interact with each other and are involved in virus replication. To understand the mechanism of this interaction in transfected and SFTSV infected mammalian cells, plasmids expressing NP and NSs protein were constructed.
2. Tomoya Morita (Lab. of Internal Medicine, D1)
"Establishment of a new right heart function test by echocardiography in dogs"
Introduction
Heart disease is important for dogs. Until now, heart function test has been focused on left heart, and right heart function has been underestimated. Right heart dysfunction has recently been shown to predict poor prognosis and symptom. Therefore, right heart function test attracts attention in human medicine. Echocardiography is the mainstay of evaluation of heart function because it is noninvasive and repeatable test. However, the quantitative assessment of right heart function using conventional echocardiography is challenging, because of the complex shape of right ventricular (RV). Recently, right heart function can be more easily measured thanks to a new echocardiography technique, called speckle tracking echocardiography (STE). STE enable to quantify right ventricular systolic performance, and this value (RV strain) has been shown to predict prognosis, severity of the patients with several heart disease. On the other hand, STE has not applied to dogs so far. Therefore, we assess the repeatability and effect of hemodynamic change on RV strain. Good repeatability in normal beagle dogs was shown in this study. RV strain decreased in acute pressure overload condition caused by infusion U46619, which is pulmonary artery vasoconstrictor. For the future, we`ll create the chronic pressure overload model dogs and measure RV strain.
3. Nyamsuren Ochirkhuu (Laboratory of Infectious Diseases, D2)
"Study on epidemiological and immunological research for intractable infectious diseases in livestock"
Introduction
Out of 339 samples collected from cattle in the Luzon island of the Philippines, 324 (95.5%), 154 (45.4%), 209 (61.6%), 140 (41.3%), and 2 (0.6%) were positive for Anaplasma marginale, Babesia bigemina, Babesia bovis, Theileria spp., and Trypanosoma evansi infections, respectively. Phylogenetic analyses showed that A. marginale 16S rRNA gene detected in this study was in a same lineage across the world, most of B. bovis RAP-1 and B. bigemina AMA-1 genes detected were in same and similar lineage from Asian isolates, and the MPSP genes of Theileria spp. belonged to the group of T. sergenti and T. buffeli.
The seroprevalence of Mycobacterium avium subspecies paratuberculosis in Mongolian cattle was examined by ELISA, and 3 (0.84%) seropositives were detected from 356 cattle. The bovine leukemia virus genome was detected by a nested PCR, and 20 (3.86%) were positive out of 517 samples. Phylogenetic analysis showed that 6 isolates were classified into genotype-4, one into genotype-7 and the other 4 into genotype-1. Bovine viral diarrhea virus was detected by immunochromatographic tests, and 6 (5.5%) were positive out of 110 serum samples. However, the virus genome was not detected by RT-PCR in the cattle positive for the BVDV antigen.
4. Wakako Furuyama (Div. of Global Epidemiology, CZC, D2)
"Host signaling pathways involved in antibody-dependent enhancement of Ebola virus infection"
Introduction
Antibody-dependent enhancement (ADE) of Ebola virus (EBOV) infection has been shown to depend on the cross-linking of virus-antibody or virus-antibody-complement complexes to cell surface Fc or complement receptors, leading to enhanced infection of susceptible cells. In this study, we examined whether the Fc receptor-dependent ADE entry of EBOV requires host cell signaling. ADE-dependent infectivity of vesicular stomatitis virus pseudotyped with EBOV glycoprotein (VSV-EBOV GP) was first compared using Jurkat cells transduced with cDNA encoding full-length Fc gamma receptor IIa (FcγRIIa) or mutant FcγRIIa lacking its cytoplasmic tail (FcγRIIa (CT-)). We found that infectivity of VSV-EBOV GP was significantly enhanced in cells expressing full-length FcγRIIa in the presence of an ADE antibody, whereas the enhancement of infectivity was limited in FcγRIIa (CT-)-expressing cells. To investigate possible signaling pathways downstream of FcγRIIa in the ADE entry of EBOV, ADE-dependent and -independent infectivities of VSV-EBOV GP were compared using K562 cells naturally expressing FcγRIIa in the presence of several inhibitors of FcγRIIa-mediated signaling pathways. We found that the ADE-dependent infection was exclusively reduced by R788 and PP2, inhibitors of Syk or Src family PTKs. These results suggest that the FcγRIIa-mediated signaling pathways are important for ADE of Ebola virus infection.
1. Huy Duc Chu (Lab. of Microbiology, DC2)
"The evaluation of hygiene status at live bird markets (LBMs) in Vietnam by virological survey and knowledge, attitude and practice (KAP) survey"
Introduction
The findings of previous study suggested that LBMs have a potential to act more than backyard farms as the primary source of AI virus as well as H5N1 HPAI virus to amplify, maintain, circulate and transmit the virus in the environment in Vietnam (Okamatsu et al., Virus Genes. 2013). It was reported that closing of LBMs was effective solution to minimize a risk of virus transmission to humans. However, closing of LBMs is a difficult challenge at the moment but it can do step-by-step. Because of cultural issues and tradition of live poultry consumption may not be changed immediately. It is important to improve biosecurity of LBMs to minimize a virus transmission to other animals and humans for the control of AI not only in Vietnam but also in Asian countries. We are conducting the virological survey and KAP survey e for the improvement of hygiene status of LBMs for the control of HPAIV. The preliminary results indicated that the AI prevalence at LBMs is quite high, 42.1%. There are 64 AI viruses were isolated including HPAIV H5N1, H5N6 strains.
2. Jemima Tiwaa Marfo (Lab. of Toxicology, D2)
"Determination of urinary neonicotinoid and its relation to some typical symptoms"
Introduction
Neonicotinoid insecticides are used worldwide in agriculture, domestic and public pest control. The Environmental health effects of six neonicotinoids and a metabolite, namely, imidacloprid, acetamiprid, nitenpyram, thiacloprid, clothianidin, thiamethoxam and N-desmethyl-acetamiprid were studied on humans. The association between urinary chlorinated-NN/metabolite and clinical symptoms in 85 subjects from the Japanese general population was evaluated by a prevalence case-control study. The detection of thiamethoxam or N-desmethyl-acetamiprid was correlated to the simultaneous exhibition of postural finger tremor, recent memory loss, headache, general fatigue, palpitation/chest pain, abdominal pain, and muscle pain/weakness/spasm with electrocardiographic findings. Their symptoms mostly continued for several days to months after prohibiting consumption of locally grown produce. The results suggest environmentally exposed NN/metabolite may cause adverse health effects on humans. As a result of the outcome of that study, quantification method for dinotefuran which is mainly used in Japan is currently under investigation. Enzymatic kinetics of neonicotinoids can also provide useful information for detoxification. Therefore, in vitro enzymatic metabolic activity measurement of some of the neonicotinoids such acetamiprid is being studied. Also, methods for other unique metabolites of neonicotinoids such as 6-chloronicotinic acid (common metabolite for imidacloprid, acetamiprid and nitenpyram) and chlorothiazolecarboxaldehyde (common metabolite for clothianidin, thiamethoxam and thiacloprid) in urine shall be developed.
"Improvement of Layperson's Understanding in Clinical Trial Using SIDCER Methodology: A Randomized-Controlled Study "
Introduction
Introduction: Informed consent form (ICF) is a required document in clinical studies. However, the quality of ICF is problematic due mainly to its extensive length with jargons and a lack of essential elements required by regulations. As such, the Strategic Initiative for Developing Capacity in Ethical Review (SIDCER) has recently proposed the SIDCER methodology for ICF development.
Objective: To evaluate the comprehension of laypersons with two interventions: SIDCER ICF vs. conventional ICF, using a post-test questionnaire as an assessment tool.
Methods: 550 laypersons in Chiang Mai, Thailand, were randomly assigned to read either SIDCER ICF (group I) or conventional ICF (group II) (1:1) and subsequently performed the post-test. The primary endpoint was the proportion of participants who have the post-test score of .80%; the secondary endpoint was the total score of the post-test.
Results: The primary endpoint was higher in group I, compared to group II (82.2% vs. 60.4%, p <0.001). The median total score in group I was higher than that in group II (19/21 vs. 18/21, p <0.001).
Conclusion: The ICF developed based on the SIDCER methodology can enhance the understanding of laypersons in this study. Evaluation of its effectiveness in different types of studies and languages is necessary.
2. Nozomi Imanishi (Nagasaki University Graduate School of Biomedical Sciences)
"Morphological and phylogenetic studies on Anopheline mosquitoes in Hokkaido"
Introduction
There was a survey to refer to this issue previously in 1968-1974, however, no definitive surveys had been carried out thereafter to approach to it systematically. In Kushiro wetland, in northern Japan, there is a newly discovered population of anopheline mosquitoes (=An. sp. Kushiro) that has a unique DNA sequence, different from any of the Anopheles species in Japan. Internal Transcribed Spacer 2 (ITS2) sequences of rDNA identified this population as Anopheles belenrae Rueda, first described in Korea in 2005. However, the taxonomic status of An. sp. Kushiro is still unclear. In this study, further phylogenetic study was performed to ask if An. sp. Kushiro is genetically identical to An. belenrae not only ITS2 region but also DNA barcoding region (partial mtDNA COI). Moreover, further morphological comparison between An. sp. Kushiro and the Korean An. belenrae was performed using paratype specimens.
3. Satoshi Shimada (Institute of Tropical Medicine, Nagasaki University)
"Therapeutic effect of post-exposure treatment with antiserum on severe fever with thrombocytopenia syndrome in a mouse model of SFTS virus infection"
Introduction
Severe fever with thrombocytopenia syndrome is an emerging viral disease that is endemic in China, Korea and Japan. No effective vaccine or specific treatment for SFTS is currently available. In this study, we examined the effects of ribavirin, site-1 protease inhibitor PF-429242, steroids, minocycline and ciprofloxacin, and antiserum from a recovered SFTS case on SFTSV infection using an in vivo mouse model. Administration of antiserum protected against lethal infection with SFTSV. This treatment protected against clinical signs of disease due to non-lethal infection. Although ribavirin had no statistically significant effect, treatment still had a slight protective effect against SFTSV infection. MC treatment resulted in prolonged survival times during lethal infection. Although other agents had no significant protective effects, they did not show detrimental effects leading to disease progression. Our results suggest that antiserum treatment may be clinically useful for post-exposure prophylaxis against SFTSV infection.
4. Amuza Byaruhanga Lucky (Institute of Tropical Medicine, Nagasaki University)
"In vitro protracted cultivation of Plasmodium knowlesi for blood stage parasitology"
Introduction
In the past decade, concerted efforts and interventions have realised a significant decrease in global malaria incidence. At the same time, Plasmodium knowlesi has gained notoriety as one of the common causes of human malaria in Southeast Asia. The salient feature of this simian parasite is the ability to establish severe disease in both humans and monkeys through natural infection. In order to understand this unusual host sharing and adaptation to a wider niche of host cells, establishment of long term P.knowlesi in vitro culture that obviates the need for animal subjects and containment facilities is necessary. Firstly, we tried to adapt monkey-derived P. knowlesi H strain using published culture conditions for this parasite. However, we were able to maintain culture up to only 4 weeks. To overcome this challenge, multiple culture medium conditions were explored to optimise long term cultivation. To this end, we successfully established an in vitro culture condition that is able to support long term P. knowlesi H strain cultivation. By using this optimised condition, we also established culture-adapted P. knowlesi Hackeri strain for the first time. Further experiments to determine genetic factors mediating P. knowlesi host switch are ongoing.
1. Kochi Toyomane (Div. of Infection and Immunity, CZC, D1)
"Investigation of the molecular mechanism of AtxA-mediated virulence regulation on Bacillus anthracis"
Introduction
Anthrax is one of the fetal infectious diseases in human and animals, caused by Bacillus anthracis which is gram-positive spore forming bacterium. B. anthracis distributes worldwide including Japan, although no anthrax cases have been reported since 2001 in this country. The pathogenicity of B. anthracis depends on two major virulence factors, the anthrax toxin and the capsule. The anthrax toxin is composed of protective antigen, edema factor and lethal factor. These factors are coded on two plasmids, pXO1 for the toxin proteins and pXO2 for capsule synthases, and coordinately regulated at the transcription level by anthrax toxin activator gene, atxA. AtxA is a pXO1-encoded transcription factor and found in B. anthracis and the few B. cereus strains that harbor pXO1-like plasmid. It is known that AtxA activates not only expression of virulence gene but also that of the numerous genes on both chromosome and plasmids, in a CO2 dependent manner. However, activation mechanism of virulence gene expression by AtxA is not well understood. The objective of my research is to find molecular mechanism of AtxA-mediated virulence regulation, for which I am planning to analyze AtxA-DNA interations.
2. Suranji Wijekoon (Lab. of Veterinary Surgery, D1)
"Inhibitory Effects of Pentosan Polysulfate Sodium on Fos-Jun/AP1 Transcriptional Regulator Complex in Canine Bone Marrow Derived Osteoclasts In Vitro"
Introduction
Rheumatoid arthritis (RA) is generally a chronic, auto immune condition in which true cures or permanent remissions are unusual with the medication at the later stage of the disease. DMARDs that stand for disease-modifying antirheumatic drugs are an important part of an overall treatment plan. DMARDS exert minimal of anti- inflammatory effect and analgesic effect. Suppression of inflammation rather than immunosuppression in the early stages of the RA can result in substantial improvements in long-term outcomes. Recent years, it was newly recognized that PPS reduce pain and inflammation on osteoarthritis. But molecular biological mechanism of PPS as anti-bone resorptive agent on osteoclastogenesis and osteoclasts function in RA is not fully understood. The purpose of the current study is to investigate the mechanisms of action of PPS on inflammatory reaction of osteoclasts in vitro and compare with Heparin and PSGAG. PPS will be directly act on intracellular signaling pathway that play a pivotal role in many essential cellular processes such as proliferation and differentiation of osteoclasts which could be seen accelerating in the inflammation of RA process to produce osteoclasts. Bone marrow derived osteoclasts were treated during 14 days of osteoclastogenic process with PPS, Polysulphated glycosaminoglycan (PSGAG) and Heparin (0.2, 1 and 5 µg/ml) after treatment of human recombinant IL-1β (10ng/ml) as an inflammatory stimulator. Gene expression of osteoclasts differentiation markers (Carbonic anhydrase, calcitonine receptors, NFATc1, MMP9, c-Fos, RANK, Cathepsin K) and transcriptional regulator activator protein-1 was measured by using quantitative reverse transcription-polymerase chain reaction. Even there is no statistical significant on gene expression of calcitonine receptor by three treatments, highest concentration of PPS, PSGAG, Heparin significantly suppress the MMP9 gene and genomic expression of NFATC by PPS and Heparin (5µg/ml). Cathepsin K gene that is the major factor necessary for osteoclastic activity was significantly suppress by all concentration of PPS. Even Fos-Jun/AP1 transcriptional regulator complex was suppressed by heparin in dose dependent manner, PPS 1µg/ml, 5µg/ml concentrations was significantly affect AP1 activity. These data suggest that PPS capable to inhibit Fos-Jun/AP1 transcriptional regulator complex which is induced in RA by inflammatory cytokines and has a complex impact on osteoclast differentiation and production of soluble mediators of bone erosion.
3. Mizuho Suzuki (Lab. of Microbiology, D1)
"Exploring the mechanisms of interspecies transmission of H2 influenza viruses and vaccine preparation for a future pandemic"
Introduction
H2N2 influenza viruses caused a pandemic in 1957, however, the viruses have disappeared after 1968. Therefore infections of H2 viruses could be a pandemic in humans due to the lack of immunity against H2 viruses. H2 avian influenza viruses are circulating in birds and H2N3 virus was isolated from pigs. H2 viruses might transmit to humans from birds via pigs same as previous pandemic viruses. In order to prepare a future pandemic, vaccine preparation and exploring mechanisms of interspieces transmission are needed. In this study, genetic and antigenic analyses were conducted to prepare vaccines. And we conducted an experimental infection of pigs with H2 avian influenza viruses to reveal mechanisms of interspieces transmission.
The HA genes of H2 viruses were classified into two lineages; North American and Eurasian. The results of antigenic analysis suggested that antigenicity of North American and Eurasian viruses are closely related despite their genetic variation. As a future plan, viruses with high yield in embryonated eggs will be selected for vaccine strains. Three avian influenza viruses were inoculated to pigs and one virus was replicated in respiratory tract. Sequence analysis and measuring binding affinity to human-type receptor will be conducted.
4. Nipawit Karnbunchob (Div. of Bioinformatics, CZC, D2)
"Use of reciprocal best hits to explore the interspecies transmission of influenza A viruses"
Introduction
Influenza A virus is a zoonotic pathogen that infects avian and mammalian hosts. Genetic reassortment between human and avian viruses in pigs is responsible for the emergence of pandemic influenza viruses. Phylogenetic analysis is a conventional approach to study transmissions of the viruses. However, the growth in the amount of viral genetic information in the database increases the size of phylogenetic trees, creating technical difficulty in detecting the interspecies transmission. In this study, we employed the reciprocal best BLAST hits algorithm to detect interspecies transmission of avian viruses in pigs. Using nucleotide sequences of the hemagglutinin and neuraminidase of avian and swine influenza viruses, our method detected one hundred five possible avian-to-swine transmissions of H1N1, H1N2, H2N3, H3N2, H5N1, H5N2, H6N6, H7N2, H9N2, H10N5, H11N6, and H11N9. Thirteen results of them are consistent with the results from scientific papers that were published previously, suggesting that our method can correctly detect interspecies transmission.
5. Takeshi Kobayashi (Lab. of Pharmacology, D2)
"Identification of responsible mechanisms for the species difference of the effect of alpha2-agonists"
Introduction
Sedative effect of XYL has a clear species difference, very effective for cattle but less effective for pigs. DEX is effective for wide species of animals. So I hypothesized that α2A-AR subtype has some roles in this species difference.
Firstly I tried to compare the strictures of α2A-AR in human, mouse, cattle and pig. And then, I simulated the binding between α2A-AR and α2-agonists in each animals. As a result, the binding pocket of DEX was TM3, 5, 6 region in every animal's α2A-AR. Same results were obtained in XYL-α2A-AR expect the pig. The binding pocket of XYL was TM3, 4, 5 region in pig α2A-AR. This result suggests that the manner of binding would be responsible for the species difference of the effect of XYL.
So I'm planning to evaluate the changes of intracellar mechanisms evoked by XYL by using the expression systems. If the α2A-AR of each animals are expressed into AtT-20 cells, which endogenously express the Gi/o coupled GPCR, I will be able to evaluate the inhibition of Ca channel, activation of K channel and decrease of cAMP evoked by XYL. These mechanisms would be responsible for species difference.
1. Nao Eguchi (Div. of Global Epidemiology, CZC, D1)
"The mechanism of inhibition of influenza virus infection by M2e-specific antibody"
Introduction
The highly conserved nature of extracellular domain of M2 protein (M2e) among influenza A viruses makes it a potential candidate for the development of a universal vaccine. In a previous study it was found that an anti-M2e antibody (r2ss23), in the absence of neutralizing activity, inhibited virus release. Interestingly, the inhibitory mechanism still remains to be elucidated. Therefore, the present study is aimed at determining the inhibitory mechanism of r2ss23.
2. Shirin Akter (Lab. of Parasitology, D2)
"Epidemiological investigation of animal reservoir for visceral leishmaniasis (kala-azar) in Bangladesh"
Introduction
More than 60% of the world's visceral leishmaniasis (VL) cases are reported from Bangladesh, India and Nepal (WHO, 2005). The true incidence of VL in Bangladesh is close to 40,000-45,000 per year. Understanding of the epidemiological role of domestic and stray dogs as reservoir host for VL in Bangladesh is still very limited which is very important to improve the current control strategies. Serological and molecular tests were performed to investigate the dogs for the presence of Leishmania infection.
3. Chihiro Kanno (Lab. of Theriogenology, D1)
"Analysis of bovine sperm motility using computer-assisted sperm analysis (CASA) for the development of new criteria of sperm motility aimed at improvement of cryopreservation method"
Introduction
Sperm motility is very important for the fertilization. Computer-assisted sperm analysis (CASA) gives us many parameters related to motility for each sperm, but those detailed data have not used effectively. We do not know what the each parameter indicates and which kind of sperm related to fertilization. The aim of present study is to develop a profitable analyzing method of CASA data for the prediction of sperm fertility.
4. Dugar Delgermurun (Lab. of Pharmacology, D3)
"Mechanism of 5-HT outflow from glomus cells in the chick aorta induced by some chemical compounds"
Introduction
Antidepressants are believed to increase the extracellular level of serotonin (5-hydroxytryptamine; 5-HT) by inhibiting its reuptake into presynaptic cell. Amphetamine derivatives are capable of acting on amine transporters and then result in monoamine release from the neurons. Endogenous H2S is one of the key factors for rat carotid glomus cells responding to hypoxia. In this study, we investigated 5-HT outflow from 5-HT containing cells in the chick aorta induced by antidepressants, amphetamine derivatives and H2S.
5. Nesta Bortey-Sam (Lab. of Toxicology, D3)
"Concentrations of polycyclic aromatic hydrocarbon (PAH) metabolites in human and domestic animal urine, and their association with carcinogenicity"
Introduction
Polycyclic aromatic hydrocarbons (PAHs) are pollutants formed during incomplete combustion of organic materials. Many PAHs are mutagenic or carcinogenic. PAHs absorbed inside the body are metabolized (liver). There, they are converted into monohydroxylated PAHs and may be subsequently conjugated with glucuronide and sulfate before being eliminated in urine or bile. The aim of this study was therefore to determine concentrations of OH-PAHs, pyrene glucuronide and sulfate in cattle urine.
1. Yinghua Yang (Lab. of Theriogenology, D1)
"The effect of bone morphogenetic protein 4 (BMP4) on in vitro growth culture of oocytes derived from bovine early antral follicles"
Introduction
Theca cells are considered as an indispensable part to a follicle. BMP4, secreted exclusively by theca cells, is a regulator during folliculogenesis. However, the effect of BMP4 on in vitro growth (IVG) culture of oocytes from early antral follicles is still unclear. In present preliminary study, the effect of 2 doses of BMP4 added to IVG culture medium was investigated.
2. Eugene Chisela Bwalya (Lab.of Veterinary Surgery, D2)
"The effects of pentosan polysulphate or polysulphated glycosaminoglycan supplemented chondrogenic differentiation medium on chondrogenic differentiation of canine bone marrow-derived mesenchymal stem cells in three-dimensional alginate cultures"
Introduction
Articular cartilage is an avascular tissue that has a limited capacity for self-renewal and repair. Currently there is no effective treatment for osteoarthritis. Pluripotent mesenchymal stem cells offer the best alternative for cartilage tissue regeneration and repair of osteoarthritic defects by autologous implantation of differentiated stem cells. The use of PPS or PSGAG and 3-D alginate beads provides an intriguing novel approach to successful cartilage tissue engineering and warrants investigating.
3. Heidi Lynn Tessmer (Div. of Bioinformatics, CZC, D2)
"Tracking Influenza Epidemics using Bioinformatics Techniques"
Introduction
Influenza is a viral disease with annual epidemics resulting in 3 to 5 million cases and between 250,000 and 500,000 deaths [WHO]. Currently, the primary method of reducing influenza's impact is the administration of an annual vaccine. This research focuses on analyzing changes in the HA component of the H3N2 strain of human influenza to locate patterns in strain content and mutation with a goal of improving the accuracy of mutation prediction leading to a more effective influenza vaccine.
4. Jing Fang (Lab. of Veterinary Surgery, D3)
"Proliferation and differentiation of cultured canine chondrocytes in different concentrations of supplemental sera as nutrients"
Introduction
Serum is commonly used as a supplementation to cell culture media. In most cases of cell cultures, serum is added to culture media at concentrations of 5% or more. This may however cause to lose some phenotypic features of cells from cells physiologically maintained in low nutrients. The objective of this study is to investigate proliferation and differentiation capabilities in cultured articular chondrocytes under various densities of nutrients conditions.
5. Mohamed Elfeky (Lab. of Biochemistry, D3)
"Effect of adiponectin on LPS - induced HMGB1 release and HMGB1- induced inflammation"
Introduction
Adiponectin predominantly secreted from adipose tissue plays an important role in various physiological processes including lipid metabolism, insulin sensitization and anti-inflammatory responses. Although there are a great deal of interest that adiponectin attenuates inflammation, however, it is not known and there is no published report regarding whether adiponectin have a role in decreasing HMGB1 release and/or signaling.
1. Paulina Duhita Anindita (Div. of Molecular Pathobiology, CZC, D2)
"Development of Antiviral Therapy against Rabies Virus"
Introduction
Rabies is a fatal disease caused by rabies virus from genus Lyssavirus, family Rhabdoviridae. Rabies virus is usually transmitted to people by rabid animal bite, mostly from dogs but sometimes involving bats. Most cases ended with death. Currently, there is no effective drug against rabies. In this study, we attempt to establish a drug screening assay in order to find compounds that possess antiviral property against rabies and then evaluate drug candidates in vitro.
2. Yusuke Izumi (Lab.of Advanced Veterinary Medicine, D1)
"Isolation and characterization of tumor endothelial cells: investigation of immunosuppressive factor"
Introduction
Recently, a few reports indicated that the morphology and genetic expression of human and mouse tumor endothelial cells (TECs) might differ from normal endothelial cells (NECs). However, there is quite limited information about immunoreaction of TECs. In my preliminary experiment, I revealed that TECs expressed PD-L1 in immunohistopathological specimens. The purpose of my study is to isolate TECs and investigate immunosuppressive factors in TECs.
3. Jeewan Thapa (Div. of Bioresources, CZC, D2)
"Molecular characterization of Mycobacterium tuberculosis complex isolates from human and animal interface in Nepal"
Introduction
Tuberculosis is a chronic bacterial infection caused by members of M. tuberculosis complex (MTC). In Nepal, TB is prevalent in human, livestock and wild animals. Although, TB in humans has been studied, relatively comprehensively, studies of TB in livestock and wild animals are still in early stage and very few with molecular works. In this study, we are planning to study TB in human-animal interface with molecular epidemiological approach.
4. Zhifu Shan (Lab. of Veterinary Hygiene, D2)
"High throughput detection of PrPSc from prion-infected cells without PK-treatment: Cell-based ELISA for novel screening method for anti-prion compounds"
Introduction
Prion diseases are neurodegenerative disorders of humans and animals that are associated with conversion of normal prion protein (PrPC) to a disease-specific isoform of prion protein (PrPSc). No effective treatments are available for prion diseases to date. It is known that PrPSc consists of proteinase-sensitive (PrPSc-sen) and proteinase-resistant (PrPSc-res). Detection of PrPSc-sen is important because of the higher prion infectivity and conversion activity than PrPSc-res. However, most previous studies on screening anti-prion compounds only detected PrPSc-res after digesting PrPC and PrPSc-sen by proteinase K. Therefore, we established a novel, cell-based ELISA for screening of anti-prion compounds, in which PrPSc in the prion-infected cells were detected without proteinase K treatment.
5. Marvin A. Villanueva (Div. of Bioresources, CZC, D2)
"Understanding the circulation of Leptospira among water buffaloes in an intensive farm setting"
Introduction
Leptospirosis is considered the most widespread zoonosis worldwide. A wide range of animals harbor this bacteria and can be transmitted to human via infected urine. Our previous results revealed the presence of pathogenic leptospires in water buffalo, an indespensable livestock in the Philippines. In this study, we tried to understand how Leptospira circulate and maintained in an intensive farm setting, in order to control/prevent further spread of this disease which is of serious public health concern.
1. Jednipit Borthong (Div. of Bioinformatics, CZC, D1)
"Analysis of bacterial population in aquatic samples using 16s rRNA fragment through an ability of metagenomics strategy"
Introduction
Aquatic bacteria play significant roles for maintenance of elemental cycles and ecological systems. In order to comprehensively study whole population of bacteria in aquatic environments, a specific tool such as a metagenomic strategy can be used. This assay let us to elucidate the bacterial population using genetic material extracted from directly environmental samples and then sequenced through the sequencer machine. The sequences were compared the identity with the reference sequences in the database for identification of bacteria. Nowadays, metagenomic strategy becomes to the effective tool for studying the whole population of bacteria in aquatic environments.
2. Yuta Sakurai (Lab.of Biochemistry, D1)
"Age-related changes in the thermoregulatory strategy in harbor seal"
Introduction
Thick subcutaneous fat layer is important to maintain body temperature for seals. However, newborn seals store very little body fat and the thickness of their fat layer is extremely thin compared to that of adults. Thus, the other mechanisms may be involved in thermoregulation. In this study, I examined whether seals possessed brown adipose tissue (BAT) that is a site of cold-induced thermogenesis in small rodents.
3. Minato Hirano (Lab. of Public Health, D1)
"Study on genomic transportation-mechanism of neural infection of flaviviruses"
Introduction
The genus Flavivirus contains over 70 members, many of which are arthropod-borne pathogens distributed widely. Neurological diseases caused by encephalitic flaviviruses are severe and associated with high levels of mortality. However, detailed mechanisms of viral replication in the brain and features of viral pathogenesis remain poorly understood. To investigate the role of viral replication during flaviviral encephalitis, I focus on the molecular mechanism of viral genome-transportation in infected neuron.
4. Ayako Ujike (Lab. of Pharmacology, D2)
"The effect of hydrogen sulfide on enterochromaffin cells"
Introduction
Hydrogen sulfide (H2S) has been recently considered as a gasotransmitter. In the gastrointestinal tract, the amount of H2S is reported to increase in some diseases, such as colitis, suggesting that H2S acts as a pathological signal. Enterochromaffin (EC) cells, one of enteroendocrine cells, sense various luminal stimuli and regulate gastrointestinal function by 5-HT release. The purpose of this study is to reveal the relation between H2S signal and EC cells.
5. Marvin A. Villanueva (Div. of Bioresources, CZC, D2)
"Understanding the circulation of Leptospira among water buffaloes in an intensive farm setting"
Introduction
Leptospirosis is considered the most widespread zoonosis worldwide. A wide range of animals harbor this bacteria and can be transmitted to human via infected urine. Our previous results revealed the presence of pathogenic leptospires in water buffalo, an indespensable livestock in the Philippines. In this study, we tried to understand how Leptospira circulate and maintained in an intensive farm setting, in order to control/prevent further spread of this disease which is of serious public health concern.
1. Chihiro Ishii (Lab. of Toxiology, D1)
"Lead poisoning in raptors from Japan and source identification using lead stable isotope ratios"
Introduction
Lead poisoning is occurred among raptors because of consuming animals that were shot with Pb bullets or shot. In Hokkaido, the use of Pb bullets for deer hunting has been prohibited since 2001, however, one eagle died due to Pb poisoning in 2013. Few areas in other parts of Japan have regulations. In this study, we measured Pb concentrations and Pb stable isotope ratio to identify the source of poisoning.
2. Yogendra Shah (Div. of Bioresources, CZC, D1)
"Diversity of Mycobacterial interspersed repetitive units-variable number of tandem repeats among Mycobacterium tuberculosis Central Asian strain family isolates from Nepalese patients"
Introduction
Tuberculosis (TB) is the major public health problem in Nepal. Several South Asian countries has reported the predominant genotype of Mycobacterium tuberculosis (MTB) belonging to Central Asian Strain (CAS) family. Mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) is a reliable and reproducible method for differentiation of MTB isolates. However, information of its utility in determining the diversity of CAS strain is limited. In the present study, we evaluate and optimized the appropriate VNTR locus combination for the detailed genotyping of MTB strains of CAS family.
3. Naoya Maekawa (Lab. of Infectious Diseases, D1)
"Expression of PD-L1 on dog tumor cells and enhancement of IFN-γ production from tumor-infiltrating cells by PD-L1 blockade"
Introduction
Programmed death 1 (PD-1), an immunoinhibitory receptor, and programmed death ligand 1 (PD-L1), its ligand, together induce the "exhausted" status in antigen-specific lymphocytes and are thus involved in the immune evasion of tumor cells. In this study, dog PD-1 and PD-L1 were molecularly characterized, and their potential as therapeutic targets for dog tumors was discussed. The final goal of this study is to establish a novel immunotherapy against dog tumors.
4. Sawa Onouchi (Lab.of Anatomy, D2)
"Morphogenesis of the murine duodenojejunal flexure"
Introduction
The gastrointestinal tract has complex flexures for its compaction to fit into the abdominal cavity. These flexures are organized for the proper digestive system and determine gut unique morphology. Mammalian animals have species-common flexures in their gut morphology, and the duodenojejunal flexure is one of them. In this study, I focus on its morphogenesis as a model for flexure formation in intestinal development.
5. Nozomu Yokoyama (Lab. of Veterinary Internal Medicine, D2)
"Expression of claudins in the colorectal mucosa of inflammatory colorectal polyps in miniature duchshund"
Introduction
Inflammatory colorectal polyps (ICRPs) were recently recognized as a major cause of large bowel diarrhea in miniature dachshunds in Japan and are thought to be a novel form of inflammatory bowel disease (IBD). In human IBD, it has been suggested that derangements in tight junction (TJ) proteins may be involved in aberrant immune response in the intestine. The aim of this study is to determine the expression and distribution of TJ proteins claudins in the colorectal mucosa of ICRPs.
1. Yuji Wada (Div. of Molecular Pathobiology, CZC, D1)
"Study on Chikungunya virus; development of novel therapy and epidemiological survey"
Introduction
Chikungunya virus (CHIKV) is the cause of chikungunya fever, reemerging infectious disease, characterized by abrupt fever, rash, and acute and chronic joint pain. CHIKV is spreading all around the world including not only tropical areas that have often suffered from various insect-mediated infectious diseases but also temperate areas such as Italy and America. Because there are no approval drugs or vaccine, it is difficult to treat patient suffered from CHIKV and to prevent CHIKV spreading.
2. Yuri Sakaki (Lab. of Radiation Biology, D1)
"The effect of downregulation of DNA repair enzyme apurinic/apyrimidinic endonuclease 1 on cellular functions"
Introduction
Apurinic/apyrimidinic endonuclease 1 (APE1) plays an important role in the repair of DNA. Recent discoveries of new functions of APE1 show that APE1 is a multifunctional protein. However, it has not been well understood how these functions of APE1 affect cellular functions as a whole. In this study, the effect of APE1 downregulation on gene expression and cellular function was evaluated to reveal the biological role of APE1.
3. Minori Kuroda (Lab. of Veterinary Hygiene, D2)
"Involvement of astrocytes in neuropathogenesis of prion diseases"
Introduction
Activation of glial cells is a hallmark in prion diseases; however, the role of activated astrocytes remains to be elucidated. Thus my Ph.D. research focuses on understanding of astrocytes involvement in pathobiology of prion diseases. I am analyzing expression activation state of astrocytes from brains of prion-infected mice. Expression of CXCL10, which has multiple functions in neurodegeneration diseases was upregulated during 90 - 120 dpi.
4. Kiyeon Kim (Div. of Bioinformatics, CZC, D2)
"Inferring the population structure of Influenza A viruses with Tajima's D and its application to surveillance"
Introduction
Influenza has been big burden to public health all over the world. And thank to active surveillance and sequencing technique we can easily get huge amount of sequence data of Influenza A virus in public. Meanwhile, Tajima's D is a statistics that explain the state of population by using sequence dataset. Using public dataset, in this study, we tried to apply Tajima's D to comprehend epidemic situation in population.
1. Teppei Ikeda (Lab. of Anatomy, D1)
"Analysis of the functions of molecules that support bidirectional functions of caspases in skeletal muscle"
Introduction
Caspases are a family of cysteine proteases that are known to regulate apoptotic signal. Apoptosis by activation of caspases is strongly associated with embryonal development and regeneration in many organs. In previous study, we detected the activation of caspases in skeletal muscle of mouse embryo, whereas there were little apoptotic cells. In this study, we focus on apoptotic and non-apoptotic functions of caspases and clarify the mechanism of myogenesis and pathogenesis that caspases activation regulates in skeletal muscle.
2. Asami Nishimori (Lab. of Infectious Disease, D1)
"A simple and rapid method for diagnosis of bovine leukemia virus (BLV) infection"
Introduction
Bovine leukemia virus (BLV) is a retrovirus associated with persistent lymphocytosis and enzootic bovine leukosis. The recent research showed that the prevalence of BLV infections in Japanese dairy cattle was increased more than 8-fold in past thirty years. In this study, we attempted the establishment of a new method which was more simple and rapid for diagnosis of BLV infection, and compared its efficacy with that of a conventional method.
3. Keigo Fukano (Lab. of Biochemistry, D2)
"The proliferative ability of mature brown adipocytes and its regulation"
Introduction
There are two types of fat tissue in mammals, white adipose tissue and brown adipose tissue. It is generally assumed that the number of white adipocyte is controlled by the proliferation of pre-adipocytes and their differentiation, and mature adipocytes have no proliferative ability. However, little is known about the regulatory mechanisms of brown adipocyte number. To clarify the proliferative ability of mature adipocyte, we generated adipocyte-specific cell-cycle arrested mice.
4. Tomoyuki Yamaguchi (Div. of Bioresources, CZC, D2)
"The role of DNA gyrase and impact of amino acid substitution in DNA gyrase"
Introduction
DNA gyrase is a bacterial enzyme, which plays an essential role in bacterial replication and transcription by giving topological changes to the double strands of DNA. This enzyme is a target of quinolone, a family of commonly used antibacterial drugs. Amino acid substitution in the quinolone-binding site may confer quinolone resistance on bacteria. I will explain how DNA gyrase works and also talk about my research work on this enzyme.
1. Balazs Oroszlany (Lab. of Toxicology, D3)
"Predictive Toxicology and modeling the reptile xenobioticmetabolism with transgenic yeast"
Introduction
Toxicology has to face two big challenges. One is to move away from animal testing (3R principles), and the other, at the same time, is to increase the throughput of pharmaceutical and chemical assays. This resulted in a fast development of human-focused *in silico* and *in vitro* screening methods. But how do these techniques translate into environmental risk assessment?
*Saccharomyces cerevisiae* based reptile xenobiotic metabolism models could answer that question.
2. Mohamed Moustafa (Lab. of Wildlife Biology and Medicine, D3)
"Epidemiology, isolation and identification of Anaplasma phagocytophilum from sika deer (Cervus nippon) in Hokkaido, Japan."
Introduction
Anaplasma phagocytophilumis a Gram-negative, obligate intracellular bacterium and causes human granulocytic anaplasmosis (HGA), which is a tick-borne rickettsial disease. In USA, the number of anaplasmosis cases reported to CDC has increased steadily since the disease became reportable, from 348 cases in 2000, to 1761 cases in 2010. Therefore, a study is required to understand the epidemiology of A. phagocytophilum and identify its strains in sika deer of Japan.
3. Siriporn Kongsoi (Div. of Bioresources, CZC, D3)
"Impact of mutations on DNA gyrase genes of Salmonella enterica"
Introduction
Quinolones are the first drug of choice for treatment of systemic salmonellosis that occurs in humans and animals. The prevalence of quinolone resistant Salmonella has become a concern for public health. Amino acid substitutions conferring resistance to quinolones in Salmonella enterica have generally been found within the quinolone resistance-determining regions (QRDRs) in the A subunit of DNA gyrase (GyrA).
1. Wakako Furuyama (Div. of Global Epidemiology, CZC, D2)
"Study on host signaling pathway involved in antibody-dependent enhancenment of Ebola virus infection"
Introduction
Antibody-dependent enhancement (ADE) is a well-known phenomenon in which viral infectivity is increased by virus-specific antibodies in vitro. Ebolaviruses utilize viral glycoprotein-specific antibodies for their entry into cells through the interaction of the antibodies with cellular Fc receptors or complement component C1q and its receptors. Since these cellular molecules is known to activate various signaling pathways, we examined whether the ebolavirus ADE requires host cell signaling pathways through these molecules.
2. Tomokazu Tamura (Lab. of Micobiology, D3)
"Extra story of my PhD study－Immunopathogenesis of classical swine fever virus infection-"
Introduction
During evolution, viruses have acquired numerous mechanisms to evade or subvert key elements of the host viral response. For classical swine fever virus, the strategy to dampen the immune response by suppression of type I interferon and RNase activity was established. However, several questions are still remaining. I will show some pieces of data that could lead to understand virus-host interaction.
3. Katsuhisa Nagai (Lab. of Theriogenology, D3)
"The relationship between antral follicle counts (AFC) and quality of oocytes in cattle"
Introduction
Ovarian reserve is defined as the capability of ovaries to produce fertilizable oocytes, and antral follicle count (AFC) in ovaries is used as an indicator. Though reduced ovarian reserve is considered to impair the quality of oocytes in human, relationship between AFC and oocytes quality in cattle is still unclear. In this study, to clarify this relationship, fertilizabilities of oocytes derived from living cows with different AFC were examined.
1. Motofumi Suzuki (Lab. of Radiation Biology, D1)
"Elucidation of the relationship between spindle assembly checkpoint and cellular radiosensitivity"
Introduction
In eukaryotes, the spindle assembly checkpoint (SAC) acts to maintain genome stability by delaying cell division until accurate chromosome segregation is ensured. Many studies have demonstrated that G1/S and G2/M checkpoints are two important checkpoints to determine the fate of irradiated cells. On the other hand, the role of SAC in cellular radioresponse remains elusive. The purpose of my study is to elucidate the relationship between SAC and cellular radioresponse.
2. Naganori Nao (Div. of Global Epidemiology, CZC, D2)
"Different pathogenicity of H5N1 highly pathogenic avian influenza viruses determined by a single amino acid in the M1 protein"
Introduction
Two highly pathogenic avian influenza virus strains, A/duck/Hokkaido/WZ83/2010 (H5N1) (WZ83) and A/duck/Hokkaido/WZ101/2010 (H5N1) (WZ101) showed different pathogenicity to chickens and ducks in spite of high nucleotide sequence identity of them. We revealed that the difference between WZ83 and WZ101 in the pathogenicity for chickens and ducks is determined only by the single amino acid in the M1 protein.
3. Keisuke Aoshima (Div. of Molecular Pathobiology, CZC, D3)
"Analysis of Reprogramming of Mouse Pronuclear Zygotes"
Introduction
Epigenetic reprogramming in zygotes is required to obtain the totipotency by erasing parental epigenetic marks such as DNA methylation and histone modifications. Although it is reported that reprogramming predominantly occurs in paternal pronucleus at the pronuclear stage soon after fertilization, the mechanism remains poorly understood. I would like to report on my progress so far in this meeting.
1. Jemima Tiwaa Marfo (Lab. of Toxicology, D1)
"Establishment of diagnosis and elucidation of toxicological effects of subacute and chronic exposure to neonicotinoids"
Introduction
Neonicotinoid pesticides are systemic insecticides that possess nicotinic acetylcholine receptor agonist activity. Their importance in protecting crops and pest insect attack has been established. Nevertheless, there are some reports about toxicity of neonicotinoids to other animals and humans.
In regards to that, my aim is to investigate the toxicity of neonicotinoid pesticides to humans.
2. Nipawit Karnbunchob (Div. of Bioinformatics, CZC, D1)
"Role of α2A-adrenoceptor for the effects of dexmedetomidine and xylazine"
Introduction
Influenza A virus is a zoonotic pathogen that infects avian and mammalian hosts. Pigs are potential intermediate hosts for viral genetic reassortment between human and avian viruses. This genetic reassortment can led to new viral strains that might cause a pandemic. Thus, monitoring influenza A viruses in pigs play an important role to control the disease. The aim of this study is to clarify inter species transmission in avian, swine and human species.
3. Huy Duc Chu (Lab. of Microbiology, D1)
"Potency of an inactivated influenza vaccine prepared from A/duck/Mongolia/119/2008 (H7N9) against the challenge with A/Anhui/1/2013(H7N9) "
Introduction
H7N9 influenza virus infection in humans was recently reported in China on March 31, 2013. Characterization of the H7N9 influenza virus indicated that it has the potential to infect humans. The urgent need for the development of such a vaccine has been acknowledged by the WHO. A/duck/Mongolia/119/2008 (H7N9) was used to prepare an inactivated whole virus particle vaccine. The potency of this test vaccine was evaluated by a challenge with A/Anhui/1/2013 (H7N9).
1. Minori Kuroda (Lab. of Veterinary Hygiene, D1)
"Involvement of astrocytes in neuropathogenesis of prion diseases"
Introduction
Prion diseases are fatal neurodegenerative diseases that are characterized by PrPSc, neuronal vacuolation and microglial activation and astrocytosis. Recent studies in my lab suggested that after the appearance of PrPSc, astrocyte activation precedes microglial activation in the early stage of prion infection; however, the role of astrocytes in prion diseases remains to be elucidated. Thus I am focusing on elucidation of roles of astrocytes in neuropathogenesis of prion diseases.
2. Takeshi Kobayashi (Lab. of Pharmacology, D1)
"Role of α2A-adrenoceptor for the effects of dexmedetomidine and xylazine"
Introduction
Dexmedetomidine and xylazine, α2-adrenoceptor agonists, are used as the analgesics or sedatives for human and animals. Descending antinociception system and α2-adrenoceptor at spinal cord may involve these effects and α2A-adrenoceptor subtype has been reported to have a important role, but the mechanisms for these drugs are still unclear. So, I estimated the effects of dexmedetomidine and xylazine by in vitro and in vivo methods using C57BL/6J and α2A -adrenoceptor knockout mice.
3. Junpei Kimura (Lab. of Anatomy, D2)
"Elucidation of the pathogenesis of glomerular disease"
Introduction
Chronic kidney disease (CKD) is one of the most serious public health problems because it is strongly associated with not only end-stage renal disease (ESRD) but also cardiovascular diseases. Although membranous proliferative glomerulonephritis (MPGN) is a major primary cause of CKD, little has been known about the pathogenesis of MPGN. Here, I investigate the genetic factors contributing glomerular injuries using MPGN model mice.
1. Ryuji Kaede (Lab. of Biochemistry, D1)
"Roles of C1q/TNF related proteins in bovine mammary gland development and mastitis"
Introduction
Milk is important nutritious drink including proteins, lipids, calcium and so on, especially as safe drink in developing countries instead of contaminated water. Bovine mastitis damages mammary glands and significantly decreases milk production. C1q/tumor necrosis factor-related proteins (CTRPs), novel adipokines, paralogs of adiponectin, play an important role in anti-inflammation and so on. In terms of mammary gland, physiological roles regulated by CTRPs are completely unknown. So, we focus on roles of CTRPs in bovine mammary glands.
2. Hidehiro Yoshimura (Lab. of Infectious diseases, D1)
"Epidemiological research of Borrelia miyamotoi in Hokkaido"
Introduction
Tick parasites impact economically by reducing animal weight gain and livestock production. Ticks also transmit many kinds of pathogens that cause diseases which greatly impact human and animal health, or zoonotic diseases. Borrelia miyamotoi (B. miyamotoi) causes relapsing fever. B. miyamotoi was firstly isolated from Ixodes persulcatus in Hokkaido in 1995.In recent years, clinical cases in human were reported in Russia and North America, and field isolates in Russia were genetically close to those in Japan. No clinical cases in Japan have been reported, but B. miyamotoi may cause future outbreak in Japan. We researched the distribution of B. miyamotoi in Ixodes ticks in Hokkaido.
1. Shan Zhifu (Lab. of Veterinary Hygiene, D1)
"Studies on the establishment of cell and immune therapy model for prion diseases"
Introduction
Bone marrow derived mesenchymal stem cells (MSCs) has been reported to show some therapeutic effects on prion infected mice. Anti-PrP antibodies or neurotrophic factors also have the neuroprotective effects on neurodegenerative diseases. Therefore, the present study is to build the MSC which can express anti-PrP antibody or neurotrophic factors and to find if they show the synergistic effects on therapy for prion diseases.
2. Fang Jing (Lab. of Veterinary Surgery, D2)
"Behavior of cultured canine chondrocytes under hyaluronic acid supplementation and low-density nutrients"
Introduction
Hyaluronic acid (HA) is a disease-modifying osteoarthritis drug to improve cartilage environment. In investigation of HA in vitro, 10% serum is commonly supplemented for maintenance of chondrocytes viability. However, chondrocytes in articular cartilage are in low-density nutrients. The aim of this study is to investigate effects of HA supplementation to culture media of chondrocytes on their proliferation, phenotype and differentiation under low-density nutrients.
3. Mohamed Sobhy (Lab. of Biochemistry, D2)
"Role of adiponectin in the regulation of obesity associated HMGB1 induced inflammation"
Introduction
Obesity is a metabolic disease of pandemic proportion. Obesity associated with a chronic inflammatory response characterized by increased expression of pro-inflammatory adipokines and decreased expression of the anti-inflammatory adipokine, adiponectin. The anti-inflammatory properties of adiponectin may be a major component of its beneficial effects on cardiovascular and metabolic disorders including atherosclerosis and insulin resistance. High-mobility group box 1 protein (HMGB1), a cytosolic ubiquitous protein stimulate necrosis-induced inflammation. Moreover, HMGB1 induces other cytokines. The identification of HMGB1 inhibitors is therefore of significant experimental and clinical interest.
1. Sawa Onouchi (Lab. of Anatomy, D1)
"Novel Theory of Gut Morphogenesis -Spatiotemporal Gene Expression Designs Flexure Formation in Fetal Mice-"
Introduction
The gastrointestinal tract has complicated morphology, containing both species-common and species-specific features, meanings that gut morphogenesis is controlled by genetic programs. In this study, we are investigating murine gut morphogenesis especially that of the duodenojejunal flexure (DJF) based on spatiotemporal gene expression, different from the common theory depending on mesenteric tension. These findings will provide a crucial knowledge in the areas of developmental anatomy as well as regenerative medicine.
2. Tatsuyuki Osuga (Lab. of Internal Medicine, D2)
"Vitamin D status in canine heart disease"
Introduction
In humans with heart failure, vitamin D deficiency is common and associated with severe heart failure symptoms and short survival. In addition, recent evidence suggests that vitamin D supplementation in heart failure patients with vitamin D deficiency may prolong their survival. In dogs with heart disease, the status of vitamin D is unknown. The purpose of the study was to evaluate the status of vitamin D in canine heart disease.
3. Tomohiro Okagawa (Lab. of Infectious Diseases, D2)
"Bovine immunoinhibitory receptors: novel therapeutic targets for bovine leukemia virus infection in cattle"
Introduction
The immunoinhibitory receptors contribute to the immune evasion mechanisms of several viruses causing chronic infections. In bovine leukemia virus (BLV) infection, several inhibitory receptors played a critical role in functional exhaustion of T cell and disease progression. We are underway to establish the monoclonal chimeric antibodies specific to bovine inhibitory receptors and investigate these effects to develop a new therapeutic method for BLV infection.
1. Delgermurun Dugar (Lab. of Pharmacology, D1)
"Mechanism of 5-HT release from glomus cells in the chick Aortic body (AB)"
Introduction
The glomus cells in chick aortic body (AB) sense changes in plasma PO2, PCO2 and pH and then release 5-hydroxytriptamine (5-HT). The purpose of this study is to reveal the effect of H2S and SSRIs drugs on 5-HT release from the glomus cells. In my preliminary experiments, we found that by both H2S and some SSRIs increased 5-HT release from the glomus cells in the chick AB.
2. Marvin A. Villanueva (Div. of Global Epidemiology, CZC, D1)
"Epidemiological study of leptospirosis among water buffalo (Bubalus bubalis)"
Introduction
Leptospirosis is a global re-emerging zoonotic infection caused by Leptospira spp. This study investigated the water buffalo, an indispensable animal in the Philippines that may contribute in maintaining and infecting leptospirosis to humans. Indirect ELISA system using recombinant outer membrane proteins were used in this study. Results showed all serum samples tested reacted to recombinant antigens used, suggesting that the disease is circulating and maintained within the herd, and may cause public health concern.
1. Ayako Ujike (Lab. of Pharmacology, D1)
"The effect of hydrogen sulfide on enterochromaffin cells"
Introduction
Hydrogen sulfide (H2S), generally known as a toxic gas, is suggested to act as a gasotransmitter in mammalian body. High concentration of H2S is detected in large intestine and H2S production is reported to increase under pathological conditions such as colitis. The purpose of this study is to examine the effect of H2S on enterochromaffin cells which are distributed in the intestines and modulate intestinal function by 5-HT release.
2. Takahiro Hiono (Lab. of Microbiology, D2)
"Analyses of influenza virus receptors on the epithelial cells of chicken trachea"
Introduction
Wild ducks are the natural hosts of influenza A viruses. Although avian influenza viruses recognize SAα2,3Gal as virus receptors, chickens are rarely infected with viruses of duck origin. In order to better understand relationship between receptor specificity and host range of influenza virus in ducks and chickens, receptor binding specificity of the viruses and the distributions of receptors in tissues of ducks and chickens were investigated.
3. Saori Yano (Lab. of Physiology, D2)
"Characteristics of estrus cycle-dependent preference for male mice's odor"
Introduction
In mammals, individual information of a donor is included in odor. Female mice obtain information about males via the odor of the urine and choose mating partner. In a previous study, we found that the preference of females for odor of particular males is changed according to estrus cycle. The aim of this study is to investigate possible mechanism underlying regulation of the odor preference.
1. Kim Kiyeon (Div. of Bioinformatics, CZC, D1)
"Inferring the population structure of viruses with Tajima's D and its application to global surveillance"
Introduction
Tajima's D is a statistic that discriminates mutation patterns in a set of nucleotide sequences. Based on the coalescent theory, Tajim's D tells us whether the population is expanding or declining and this property can be used for inferring the change of demography and evolution of population.
2. Bortey-Sam Nesta (Lab. of Toxicology, D1)
"Prevalence of Polycyclic aromatic hydrocarbons (PAHs) and their Metabolites in air, soil and urine of humans and domestic animals from Kumasi-Ghana"
Introduction
Polycyclic Aromatic Hydrocarbons (PAHs) are environmental pollutants found in air, soil, water and food. Most PAHs entering the environment are formed during burning of fossil fuel, wood etc. Many PAHs are mutagenic or carcinogenic. The aim of this study is to compare concentrations and abundances of PAHs and their metabolites (OH-PAHs), their contribution to the overall carcinogenic potential of PAH mixture and to assess the major sources of PAHs pollution in Kumasi.
3. Sarad Paudel (Lab. of Wild life Biology and Medicine, D2)
"Epidemiological and molecular studies of tuberculosis in captive elephants of Nepal"
Introduction
Epidemiological and molecular studies of tuberculosis (TB) were conducted in captive elephants of Nepal. Serological testing has shown that 25% elephants were infected with TB. Mycobacterium tuberculosis was isolated from three elephants having suspected TB lesions upon necropsy. Genotyping of isolates showed that the elephants were infected with local Mycobacterium tuberculosis strains prevalent among local population of Nepal. The results strongly suggest the transmission of TB from human to elephant.
1. Keigo Fukano (Laboratory of Biochemistry, D1)
"The mechanism of the disappearance of Brown Adipose Tissue (BAT)"
Introduction
BAT is one of two types of adipose tissue found in mammals. Its primary function is to generate body heat in animals or newborns that don't shiver. In human, it is known that the metabolic activity of BAT differs among individuals, being lower in older and obese individuals. But it is not known how it disappears. The aim of this study is to elucidate the mechanism of the disappearance of BAT.
2. Nozomu Yokoyama (Laboratory of Veterinary Internal Medicine, D1)
"The molecular mechanism and functional role of Toll-like Receptor in Canine Inflammatory Colorectal Polyps"
Introduction
Canine Inflammatory Colorectal Polyps (ICRP) in miniature dachshunds are recently recognized in Japan as a major cause of large bowel symptoms in this breed. In human, accumulating evidence indicate that Toll-like Receptors (TLRs) in intestinal epithelial cells play important roles in the pathogenesis of human IBD. The aim of my study is to examine the molecular mechanism and functional role of TLRs in the colonic epithelial cells of ICRP.
3. Junki Maruyama (CZC, Division of Global Epidemiology, D2)
"Characterization of the novel filovirus, Lloviu virus"
Introduction
Lloviu virus (LLOV) is a novel filovirus detected from bats in Europe. This virus is phylogenetically distinct from other filoviruses in the genus Ebolavirus or Marburgvirus, thus belongs to the newly proposed genus Lloviu cuevavirus in the family Filoviridae. Filovirus has a single envelope glycoprotein (GP) that plays major roles in the replication cycle and pathogenicity. In this study, we focused on GP to estimate the biological property of LLOV.
1stKatsuhisa Nagai（Laboratory of Theriogenology, D2）
【Title】
「Ovarian follicular reserve of cattle」
【Introduction】
A term "ovarian follicular reserve" is used to explain the capacity of the ovary providing oocytes with fertilizability resulting in a successful pregnancy. The number of small antral follicles in ovaries and anti-mullerian hormone produced by granulosa cells of growing follicles, both could be used as a marker of ovarian follicular reserve. So, we examined the utility of markers and the relationships between ovarian follicular reserve and fertility in cattle.
2ndYongjin Qiu（CZC, Division of Collaboration and Education, D3）
【Title】
「Tick microbial population analysis using 16S rRNA gene analysis with next generation sequencer」
【Introduction】
Ticks can transmit various pathogens. For example, nine spotted fever rickettsia species were identified as emerging agents recently. This fact indicates that ticks contain a large number of potential pathogens which have not been identified so far. 16S rRNA gene analysis is able to find unculturable microbes in environmental samples. The aim of this study is to analyze the tick microbes including potential pathogens by 16S rRNA gene analysis
1stMohamed Abdallah （Laboratory of Wildlife Biology and Medicine, D2）
【Title】
「Implications of microparasites coexistence on the infection risk in Hokkaido wildlife」
【Introduction】
The study of tick borne pathogens (TBPs) in wildlife is extremely important because more than 70% of human emerging infectious diseases (EID) between 1940 and 2004 have their origin in wildlife. We believe that the coexistence of these pathogens at the same ecological system has created a network of interactions and co-infections between them inside the hosts. Our study will include the molecular diagnosis of different TBPs in Hokkaido wildlife.
2ndNaganori Nao （CZC, Division of Global Epidemiology, D2）
【Title】
「Mechanisms for the acquisition of high pathogenicity by avian influenza viruses」
【Introduction】
It is known that highly pathogenic avian influenza viruses (HPAIVs) evolve from low pathogenic precursor viruses maintained in wild aquatic birds, the natural host of influenza A viruses. While viruses of 16 hemagglutinin (HA; H1-H16) subtypes have been identified in the waterfowl reservoirs, the HA subtype of naturally isolated HPAIVs are restricted to only H5 or H7. The purpose of our research is to identify this restriction factors.
3rdJun Moriwaki （Laboratory of Wildlife Biology and Medicine, D2）
【Title】
「Development of studies on reproduction and behavior of brown bears with a goal of conservation of biodiversity」
【Introduction】
The Hokkaido brown bear (Ursus arctos yesoensis) is the largest terrestrial mammal in Hokkaido, Japan. Human-bear conflicts have become a social problem. For the conservation and management of wild brown bears, it is necessary to understand biological traits in brown bears, including population dynamics, etc. One of the biological factors which directly affect population dynamics is reproduction. For clarifying the reproductive rate of Hokkaido brown bears, we conducted two approaches.
1stChukwunonso Onyemaechi Nzelu (Laboratory of Parasitology, D2)
【Title】
「Survey of natural infection by Leishmania species in anthropophilic sand flies」
【Introduction】
Leishmaniasis is a vector-borne disease caused by the protozoan of genus Leishmania. The protozoan is transmitted by the female sand fly. Nearly 1,000 sand fly species has been described, but only a few are medically important. The present study employed the use of molecular biological method to evaluate natural Leishmania infection rates in man-biting sand flies.
2ndSangho Kim （Laboratory of Veterinary Surgery, D2）
【Title】
「Evaluate of association between the endoplasmic reticulum stress and canine osteoarthritis」
【Introduction】
Molecular mechanism of osteoarthritis (OA) is unclear, hence, therapeutic target of OA is not established. In this study we intend to confirm the hypothesis that excessive endoplasmic reticulum (ER) stress inducing cellular apoptosis plays a role in the pathology of cartilage degeneration. So, we investigate about evaluate of association between the ER stress and canine osteoarthritis.
3rdMakoto Kuroda （CZC, Division of Global epidemiology, D2）
【Title】
「Search for a receptor for filovirus」
【Introduction】
While several cellular molecules have been proposed to be filovirus receptors or co-receptors, the molecular mechanism of filovirus entry has not been fully understood. A monoclonal antibody M224/1, which was produced against some cellular molecule on the surface of Vero E6 cells, blocks infection with vesicular stomatitis virus pseudotyped with filovirus glycoprotein. In this study, we identified the target molecule of M224/1 by an expression cloning method using Vero E6 cDNA library.
1st木村 純平 (解剖学教室, D1)
【Title】
「Analysis of Podocyte Injury in Murine Glomerulonephritis」
【Introduction】
Podocytes are highly differentiated cells lining the outside of glomerular capillaries. The podocyte foot processes form the slit diaphragm contributing to the regulation of glomerular filtration barrier. The effacement of foot processes are representative pathological features of podocyte injuries causing glomerular diseases with albuminuria. Although choronic glomerulonephritis (CGN) is a serious glomerular disease leading dialysis, little is known about the podocyte injuries. In the present study, we analyzed the pathological features of CGN model mice, especially focusing on the podocyte injuries.
2nd田村 友和 （微生物学教室, D1）
【Title】
「Identification of viral factors responsible for the pathogenicity of classical
swine fever virus in pigs」
【Introduction】
Numerous classical swine fever (CSF) virus strains show differences in CSF pathogenesis, resulting in clinical disease with different degrees of severity. The present study demonstrated that the amino acid residues in a structural protein and nonstructural proteins of CSF virus could function in a synergistic manner to determine the pathogenicity of the virus in pigs.
1stRuchirada Changkwanyeu (CZC Division of Global Epidemiology, D2)
【Title】
「Inhibition activity of quinolones against DNA gyrase of Campylobacter
jejuni」
【Introduction】
Quinolones are the antimicrobials most commonly used for treatment of adults with Campylobacter infections. Target of this drug in C. jejuni has been shown to be DNA gyrase. To investigate the interaction of C. jejuni gyrase with quinolones, we reconstituted in vitro the C. jejuni DNA gyrase by expressing the GyrA and GyrB subunits and evaluated the interaction of the enzyme with a large panel of quinolone with inhibition of DNA supercoiling assay..
2ndTatsuyuki Osuga （Lab. Of Internal medicine, D1）
【Title】
「Evaluation of left atrial phasic function by echocardiography in canine heart
disease」
【Introduction】
The left atrium modulates left ventricular filling through its 3 phasic function (reservoir, conduit, and booster pump function) to maintaining an optimal performance of the heart. In humans with heart diseases, the left atrial phasic function has been evaluated mainly by echocardiography and a strong correlation between left atrial dysfunction and the severity or the prognosis of the heart diseases has been described. In dogs with heart disease, the evaluation of the left atrial phasic function has not been reported and therefore the clinical usefulness of the assessment of the left atrial phasic function remains unknown. The purpose of the present study was to evaluate the clinical value of the assessment of the left atrial phasic function via echocardiography in canine heart disease.
3rdSayuri Nakamura （Lab. Of Comparative pathology, D2）
【Title】
「Pathologic features of astrocytic growth early after infection with fowl
glioma-inducing retroviruses」
【Introduction】
Fowl glioma is histopathologically characterized by multiple nodular astrocytic growths. This disease is caused by subgroup A of avian leukosis viruses, including fowl glioma-inducing virus (FGV). However, how and when astrocytes show neoplastic growth in this disease is not fully elucidated. Here, we investigated the relationship among viral replication, host immune responses and brain lesions in the initial infectious phase.
1stSiriporn Kongsoi (CZC Division of Global Epidemiology, D2)
【Title】
「Studies on the drug resistance in Salmonella Typhimurium, Salmonella
Enteritidis and Salmonella Choleraesuis」
【Introduction】
Salmonella gastroenteritis is generally a self-limiting illness; however, antimicrobial may be required in invasive or severe cases that may occur in some patients, particularly in vulnerable groups such as young children, elderly, and immune-compromised. Quinolones are the drug of first choice for treatment of systemic salmonellosis that occurs in humans and animals. However, resistance to these drugs has increased significantly in recent years.
2nd千原正尚 （解剖学教室, 博士課程2年）
【Title】
「Elucidation of the function of blood-testis barrier」
【Introduction】
In adult testes, differentiating meiotic and postmeiotic germ cells are separated from blood by the blood-testis barrier (BTB) formed by tight junctions between adjacent Sertoli cells. Although BTB has been considered to be essential for normal spermatogenesis, the function of BTB remains obscure. Here, we clarified the expression of BTB component proteins in adult mouse testis and investigated the spermatogenesis after BTB disruption.
3rd宮園耕介 （臨床分子生物学教室, 博士課程2年）
【Title】
「The elucidation of the alternative control mechanism of γ globin in bovine
globin switching」
【Introduction】
Globin switching in the birth period is a physiological response in which the globin chain substitution of the fetal γ-chain with the adult γ-chain. Bovine γ- and γ-globin possess the promoter sequences that are very similar to each other and the switching mechanism remains unknown. The purpose of the present study was to identify transcription factors and to analyze the mechanisms for differential promoter activation of γ- and γ-globin genes in cattle.
第1演題 Oroszlany Balazs (Laboratory of Toxicology)
【タイトル】
First steps in investigating the reptile xenobiotics metabolism: The metabolism of pyrene in freshwater turtles
【要旨】
Reptiles are one of the most endangered, and, at the same time, the most understudied group of vertebrates. To start investigating the xenobiotics metabolism in reptiles, as a first step, I have chosen freshwater turtles. Initial results indicates that turtles produce an unique metabolite distribution, which might be explained by the measured low activity of phase II enzymes.
第2演題 岡川朋弘（感染症学教室）
【タイトル】
ウシの免疫抑制受容体に対する阻害剤の開発および牛白血病ウイルス感染症の新規制御法への
応用
Investigation of immunoinhibitory receptors as novel therapeutic targets for bovine leukemia virus infection
【要旨】
牛白血病ウイルス (BLV) 感染症は、病態の進行に伴う免疫抑制を特徴とする慢性感染症である。我々はこれまでに、ウシの免疫抑制受容体を４種類同定し、これらの受容体がBLV感染症における免疫抑制に関与していることを報告した。本研究では、BLV感染症に対する新規治療法を開発することを目的に、これらの受容体を阻害するキメラ抗体を作製し、その治療効果を検討する。
Bovine leukemia virus (BLV) infection is characterised by immune supression in line with disease progression. Until now, we identified four immunoinhibitory receptors from cattle. Interestingly, we found that these inhibitory receptors play a critical role in immune supression during BLV infection. In this study, we establish the monoclonal chimeric antibodies specific to bovine inhibitory receptors, and investigate their effect to develop a new therapeutic method for BLV infection.
第3演題 足立真実（外科学教室）
【タイトル】
犬の血管肉腫転移に伴う血管内皮前駆細胞の変動
Functional changes in endothelial progenitor cells associated with metastasis of canine hemangiosarcoma
【要旨】
犬の血管肉腫は、高率に転移を引き起こす悪性腫瘍である。しかし、その転移機構は未だ解明されていない。我々は血管肉腫の転移は血管内皮前駆細胞によって引き起こされていると仮定し、血管肉腫罹患犬の末梢血中の血管内皮前駆細胞を検出することを目的として、これまでに得た成績を報告する。
Canine hemangiosarcoma (HSA) is a progressive malignant neoplasm because of a high metastasis. However, the underlying mechanism for this metastatic behavior of this tumor has not been elucidated yet. I hypothesized that this metastatic behavior was caused by EPCs. In this study, I am planning to detect circulating EPCs in the peripheral blood of dogs with HSA, and here we report some results.
dai
 |
 |
| ● Date: | March 4 (Fri), 2016, 18:00 - 20:15 |
| ● Venue: | Lecture Hall |
1. Bongkot Soonthornsata (Unit of Risk Analysis and Management, CZC, D1)
"Transmission risks of major mosquito-borne diseases in Thailand"
Introduction
Mosquito-borne diseases are causing a substantial burden of mortality, morbidity and economic loss in many parts of the world including Thailand and its neighboring countries. Given the persistent of case notifications of dengue and chikungunya including a recent uncovered Zika cases in Thailand, in combination with the abundance and presence of their significant vectors, Aedes aegypti and Aedes albopictus in the entire country, high level of population movement along border areas and within the country, and changes in environment, indicating that transmission in endemic areas will likely be continued and it is possible that transmission potential in new risk areas where suitable vectors are present will be identified. The current research is thus to investigate transmission risks of dengue and other relevant diseases in Thailand by conducting entomological and epidemiological surveillances and all data collected will be statistically assessed. The study will be taken place at 13 sites throughout Thailand and all study sites intend to start conducting their first entomological surveillance in February 2016 and mosquito samples are expected to collect three times a year. Mosquito samples collected will be used for molecular detection and genotyping of pathogens in mosquitoes and/or for monitoring of currently used insecticide resistance status.
2. Tomoya Morita (Lab. of Internal Medicine, D2)
"Assessment of right heart function by echocardiography in dog models of acute pulmonary hypertension and dogs with heart disease."
Introduction
Right heart dysfunction has recently been shown to predict poor prognosis and symptom in patients with heart disease. Therefore, right heart function test attracts attention in human medicine. However, the quantitative assessment of right heart function using noninvasive method, such as echocardiography, is challenging because of the complex shape of right ventricular (RV). Tei index is an echocardiographic index of the global myocardial function, including systolic and diastolic performance. Tei index is unaffected by complex shape of RV, and it has been shown to predict the prognosis, severity of the human patients with heart disease. However, few reports are available on the relationship between Tei index and hemodynamic variables or prognosis in dogs. Therefore, we measured Tei index in dog models of acute RV pressure overload and client-owned dogs with heart disease. As a result, Tei index reflected hemodynamic deterioration and diastolic dysfunction in dog models of acute RV pressure overload, and it was a good predictor of death in dogs with heart disease. For the future, we`ll create dog models of chronic RV pressure overload, and validate the usefulness of Tei index in the assessment of right heart function.
3. Memi Muto (Lab. of Public Health, D2)
"Isolation and characterization of tick-borne encephalitis virus from Ixodes persulcatus in Mongolia in 2012."
Introduction
Tick-borne encephalitis virus (TBEV) is a zoonotic virus belonging to the genus Flavivirus, in the family Flaviviridae. The virus, which is endemic in Europe and northern parts of Asia, causes severe encephalitis. Tick-borne encephalitis (TBE) has been reported in Mongolia since the 1980s, but details about the biological characteristics of the endemic virus are lacking. In this study, 680 ticks (Ixodes persulcatus) were collected in Selenge aimag, northern Mongolia, in 2012. Nine Mongolian TBEV strains were isolated from tick homogenates. A sequence analysis of the envelope protein gene revealed that all isolates belonged to the Siberian subtype of TBEV. Two strains showed similar growth properties in cultured cells, but their virulence in mice differed. Whole genome sequencing revealed only thirteen amino acid differences between these Mongolian TBEV strains. Our results suggest that these naturally occurring amino acid mutations affected the pathogenicity of Mongolian TBEV. Our results may be an important platform for monitoring TBEV to evaluate the epidemiological risk in TBE endemic areas of Mongolia.
4. Wallaya Phongphaew (Division of Molecular Pathobiology, CZC, D2)
"Valosin containing protein plays an important role in West Nile virus infection"
Introduction
The Endoplasmic Reticulum is related to the genome replication and assembly steps of WNV life cycle. The Valosin containing protein (VCP/p97) is a one component of ER-associated degradation (ERAD) pathway involving in transportation of ubiquitinated protein to proteasome for degradation and the other cellular processes. Previous studies revealed that VCP also participates in the life cycle of several viruses such as Poliovirus and Sindbis virus. The present study demonstrates the role of the VCP in WNV infection. Inhibition of VCP by treatment with either Eeyarestatin I or 3,4-Methylenedioxy-β-nitrostyrene inhibits the WNV infection in a dose-dependent manner. Furthermore, the depletion of the endogenous VCP expression using siRNA significantly attenuates the WNV infection. Therefore, we conclude that the VCP is required for WNV infection. We also examine which step of WNV life cycle should be participated by VCP. The results suggest that VCP may play a role in the either WNV genome replication or protein synthesis.
5. Tapiwa Lundu (Laboratory of Public Health, D2)
"Studies on the ecology of Severe Fever with Thrombocytopenia Syndrome Virus and the analysis of the mechanisms of virion formation of SFTSV"
Introduction
1) A variety of animals are suggested to be reservoir hosts of severe fever with thrombocytopenia syndrome virus (SFTSV). In this study we screened 315 deer sera from Hokkaido (non-endemic area), 41 deer sera from Miyazaki (endemic area), 4 wild boar sera from Nagasaki and 910 rodent sera from six locations in the endemic area of Japan. All the sera from Hokkaido showed a negative result by ELISA (OD<0.1). Fourteen rodent sera had slightly higher OD values on ELISA (0.1
 |
 |
| ● Date: | February 5 (Fri), 2016, 18:00 - 19:15 |
| ● Venue: | Lecture Hall |
1. Tatsunari Kondoh (Division of Global Epidemiology, CZC, D1)
"Host factors involved in the cell susceptibility to filovirus"
Introduction
1) Niemann-Pick C1 (NPC1), a protein involved in filovirus glycoprotein (GP)-mediated membrane fusion during the virus entry into cells, is believed to be a major determinant of cell susceptibility to filovirus infection. To analyze polymorphism of human NPC1s, we cloned NPC1 genes from permissive and non-permissive cell lines for filovirus infection and detected nonsynonymous polymorphism in these NPC1s. Three amino acid differences were found between NPC1s of permissive and non-permissive cell lines. We hypothesize that this polymorphism may affect efficiency of filovirus entry. In the future, we will generate cell lines expressing each NPC1 to assess the GP-dependent entry.
2) It has been reported that some non-retroviral RNA virus genes are integrated into vertebrate genomes. Recent studies suggest that expression of endogenously encoded bornavirus-like nucleoprotein confers resistance to the virus infection by inhibiting its replication in cells. Endogenous filovirus-like elements (EFLEs) have also been discovered in several mammalian genomes, however the role of EFLEs is not known. We constructed plasmid expressing an endogenous filovirus VP35-like element (mlEEL35) containing nearly full-length genes corresponding to the VP35 gene. We will express the mlEEL35 in cell culture and clarify whether it has a VP35-like function (i.e., inhibition of interferon response).
2. Rabin Kadariya (Lab. of Wildlife Biology and Medicine, D1)
"Genetic Variation, Ecology and Conservation of Asiatic Black Bears (Ursus thibetanus) in Annapurna Conservation Area of Nepal"
Introduction
The Asiatic black bear is endangered in mountain landscape of Nepal but has never been received for conservation importance. Retaliatory killing, poaching, climate change and habitat degradation have led to dwindling and scattered bear populations, however, no study on population genetic structure and ecology was conducted. Genetic variation may reduce in small breeding population due to inbreeding and genetic drift that impede adaptability of population to new environment. This situation is true in spatially isolated small bear population in Nepal. It may lead to extirpation of certain populations if timely conservation measures are not undertaken. This research explores genetic variation and seasonal diet preference of isolated bear population using non-invasive method.
During initial survey, 25 fecal and 10 hair samples were collected from bear habitat. DNA from fecal and hair samples was extracted using QIAamp mini-stool kit and DNA extractor FM kit (Wako) respectively. The highly variable region of mitochondrial DNA control region and nuclear DNA microsatellite will be sequenced for study of phylogenetic relationship and genetic variation respectively. The percent frequency of occurrence shows that wild fruits/nuts (85%) are major food followed grasses (43%) whereas small amount of mammal, insects and crop are also recorded in bear feces.
3. Suranji Wijekoon (Lab. of Veterinary Surgery, D2)
"Osteoclast precursor as cellular biomarker to assess relative protection against bone erosion in concurrent patellar luxation and cranial cruciate ligament rupture among dogs"
Introduction
Inflammatory joint disease characterized by abnormal synovial proliferation and destruction of articular cartilage and bone. Synovial membranes (SM) were collected from dogs associated with cranial cruciate ligament rupture (CCLr, n=5) and medial patella luxation (MPL, n=5). Samples were homogenized to extract RNA and another part was cultured in the presence of receptor-activator for nuclear factor-kB ligand (RANKL) and monocyte-colony stimulating factor (M-CSF) in the presence/ absence of interleukin-1β (IL1β), tumor necrosing factor-α (TNFα) and IL17 (10ng/ml). Osteoclasts differentiation was assessed by expression of tartrate-resistant acid phosphatase (TRAP), carbonic anhydraseII (CA), matrix metalloproteinase-9 (MMP9), RANK and cathepsinK (CatK). Relative expression of CA, MMP9, RANK and CatK were increases in TNFα treated cells (p<0.05) with CCLr. Cells from CCLr and MPL increased the expression of CA and RANK with IL1β (p<0.05) and TNFα (p<0.05) respectively. In the dogs with CCLr, MMP9 and RANK expression were increased (p<0.05), when compared with MPL derived samples. There was mild expression level of RANK in fresh SM from CCLr and MPL though catK expression increased in CCLr. The results of this study are expected to be emphasis the important of investigating of biomarkers and biotherapies targeting osteoclasts precursors those are presenting in vary number in CCLr and MPL to minimize joint inflammation and structural damage.
 |
 |
| ● Date: | January 8 (Fri), 2016, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Lai Lai San (Division of Bioresources, CZC, D2)
"MIRU-VNTR Typing of Beijing multidrug resistant tuberculosis strains from Myanmar"
Introduction
Myanmar is one of the countries with the highest burden of tuberculosis (TB) as well as multidrug-resistant tuberculosis (MDR-TB). The previous studies on genotping in Myanmar pointed out that the Beijing strain has a significant association with MDR-TB. The active transmission of MDR Mycobacterium tuberculosis (MTB) strains is the emerging problem, hence, it is necessary to identify characteristics that can explain why they keep spreading widely. A total of 212 Beijing MDR-MTB isolates collected from Myanmar was analyzed by variable number of tandem repeat(s) of mycobacterial interspersed repetitive units (MIRU-VNTR) typing. Fifteen loci of MIRU-VNTR ( QUB26, Mtub21, MIRU31, MIRU10, QUB11b, MIRU26, MIRU4, MIRU39, MIRU40, Mtub4, QUB4156, Mtub30, MIRU39, ETR A, MIRU16) were used and the results were analyzed by bionumeric software. Thirty-eight clusters in 212 isolates with the biggest cluster consisted of 8 isolates were found. VNTR set used in this study was useful for the discrimination of this genotype and effective for the determination of transmission pattern of MDR-TB. Beijing MDR-MTB was highly diverse and it can be inferred that the outbreak of MDR-TB is less likely to occur. MDR-MTB may be emerging in each patient individually. Therefore, we can stop spread of MDR-MTB with proper drug usage.
2. Tatsuya Deguchi (Lab. of Veterinary Surgery, D1)
"Analysis for radioresistance of cancer stem-like cells derived from canine tumors"
Introduction
CSCs have been defined as a small population of cancer cells that are resistant to radiotherapy and their survival after radiotherapy has been linked to recurrence and metastasis. Various studies have been performed to characterize human CSCs and in the canine existence of CSCs has been reported. However, it is uncertain on the level of radioresistance of these canine CSCs. My research is to identify and characterize canine CSCs and the level of radioresistant and how they relate to prognosis. In this study, CSCs of various canine cancer cell lines are identified by using sphere formation assay. The stemness of spheroid cells derived from canine cancer cells was evaluated by expression of stemness markers, tumorigenesis and investigated radioresistance. Spheroid cells from canine osteosarcoma cell line were identified. Spheroid cells showed expression of stemness markers CD133 gene. In future, spheroid cells will be investigated for expression of other stemness markers and radioresistance evaluated and compared with adherent-cultured cells. The expression of CD133 marker would suggest that spheroid cells possess stemness and provide a CSCs research tool to investigate CSCs of canine tumour cells.
3. Ochirkhuu Nyamsuren (Lab. of Infectious Disease, D3)
"Study on epidemiological and immunological research for intractable infectious diseases in livestock"
Introduction
Out of 339 samples collected from cattle in the Luzon island of the Philippines, 324 (95.5%), 154 (45.4%), 209 (61.6%), 140 (41.3%), and 2 (0.6%) were positive for Anaplasma marginale, Babesia bigemina, Babesia bovis, Theileria spp., and Trypanosoma evansi infections. Phylogenetic analyses showed that A. marginale 16S rRNA gene was same lineage across the world, most of B. bovis RAP-1 and B. bigemina AMA-1 genes detected were same and similar lineage from Asian isolates, and the MPSP genes of Theileria spp. belonged to the group of T. sergenti and T. buffeli. On the other hand, molecular epidemiological studies of several infectious diseases were done in Mongolian livestock, 3 (0.84%), 20 (3.86%), 11(8.7%) and 97 (23.2%) were positive for Mycobacterium avium subspecies paratuberculosis, Bovine leukemia virus, Bovine viral diarrhea viruses and Anaplasma marginale in cattle samples by ELISA, nested PCR and RT-PCR assays. In addition, Anaplasma ovis was detected in other ruminant species and infection rate was 3 (33.3%) of 100 yak, 189 (94%) of 201 sheep and 73 (36.5%) of 200 goat samples, respectively. The phylogenetic analysis of Bovine leukemia virus, Bovine viral diarrhea viruses Anaplasma marginale and Anaplasma ovis were performed and compared with other isolates from several countries.
4. Mizuho Suzuki (Lab. of Microbiology, D2)
"Exploring the mechanisms of interspecies transmission of H2 influenza viruses and vaccine preparation for a future pandemic"
Introduction
H2N2 influenza viruses caused a pandemic in 1957, but have not been detected from human population since 1968. H2 influenza viruses might be re-introduced into humans in the future because the H2 influenza viruses are still circulating in birds. Since most of human population are immunologically naive to H2 influenza viruses, it would cause a pandemic. Previous studies revealed that human pandemic influenza viruses originated from avian influenza viruses and emerged via pigs. The aim of this study is to evaluate pandemic potential of avian H2 influenza viruses based on their receptor specificity, and to prepare a vaccine for a future pandemic. During three-time passages of A/duck/Hong Kong/278/1978 (H2N9) in pigs, the recovered viruses did not show specificity to human-type receptors. The H2N9 influenza viruses will be passaged a few more time to clarify the pandemic potential of avian H2 influenza viruses. In order to prepare a vaccine, we analyzed avian, human and swine H2 influenza viruses genetically and antigenically. Antigenicity of H2 influenza viruses did not differ each other despite their genetic diversity. Based on this result, we will select vaccine candidate strain which growth effectively in chicken embryonated eggs and possess enough immunogenicity in mice.
 |
 |
| ● Date: | December 4 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Jinxi Wang (Lab. of Experimental Animal Science, D2)
"Studies of the Influence of Genetic Background on the Severity of Hirschsprung's Disease by Using Rat Models"
Introduction
The Hirschsprung's Disease (HSCR) is an inherited disease controlled by multiple genes with complicated genetic mechanism. Patients with this disease suffer various extent of constipation including complete intestinal obstruction. Up to now, although more than 11 genes have been identified as responsible for this disease, the vast majority (about 80%) of HSCR heritability is still hidden or missing. The Ednrb gene is one of the most important genes for the development of HSCR. By studying three different rat strains carrying the same Ednrb mutation that show a big difference in their symptom, we found 17 SNPs in the critical genomic region responsible for the different severity in mutated Ednrb gene-induced HSCR. To confirm our idea and identify the specific SNP/SNPs, we selected another two rat strains each possessing the same haplotype as rat strains showing mild and severe symptoms, respectively, and induce a mutation on their Ednrb gene by using GONAD (Genome-editing via Oviductal Nucleic Acids Delivery) method. Through this investigation, we expect to find out some hidden genetic mechanisms of the HSCR, which will help further studies of this disorder.
2. Md. Abdul Masum (Lab. of Anatomy, D1)
"Vascular associated stem cell niche and its pathological alternation in mouse kidney"
Introduction
In this study, the author firstly examined the alternation of blood vessel structures in the kidney of CKD model mice. BXSB/MpJ-Yaa mice clearly developed glomerulonephritis with tubulointerstitial lesions. B- and T-cells were infiltrated to both glomerulus and tubulointerstitium, and macrophages were diffusely found in peri-glomerular or tubulointerstitial regions. These cell infiltrations were severer at late stage compared to early stage, and significant increase was detected in B-cells (P < 0.01). IL-1F6+ injured renal tubules were more abundantly observed at late stage compared to early stage. No myofibroblasts were found in tubulointerstitial lesions at both stages. For vasculature, no significant difference was observed in the number of peri-tubular capillaries contacting with renal tubules between early and late stage. However, CD34+ peri-tubular capillary/unit area in the kidney was significantly decreased at late stage compared to early stage (P < 0.01). In examined parameters, the decreased CD34+ peri-tubular capillaries were significantly correlated with the increased B-cells (P < 0.001, ρ = -0.812). Genetic mutation in Y chromosome of BXSB/MpJ-Yaa mice causes autoimmune disease phenotypes, especially systemic increase of autoreactive B-cells. Our results indicated the negative correlation between B-cell infiltrations and decreased CD34+ capillaries with the progression of CKD. From the results of histoplanimetry, the author considered that altered microvascular environment by inflammation might affect CD34 expression in endothelial cells without the quantitative changes of capillaries.
3. Shirin Akter (Lab. of Parasitology, D3)
"Serological and molecular survey of Leishmania infection in stray dogs of visceral leishmaniasis endemic areas in Bangladesh"
Introduction
Transmission of VL caused by Leishmania Donovani is considered to be anthroponotic (human to human transmission) in the Indian subcontinent. L. infantum which is phylogenetically most related to L. donovani is found in the Mediterranean, Middle East, Asia, and South American countries, and responsible for zoonotic VL, with dogs as the main reservoir host. However, limited studies have been conducted to investigate the role of dogs in VL caused by L. donovani. In 2012, a total of 50 stray dogs in VL endemic areas of Bangladesh were screened for Leishmania infection. Anti-Leishmania antibodies were detected in 6 (12%) dog serum samples using rK39 immunochromatographic stick test (ICT). Five (10%) samples were positive for Leishmania DNA using ITS1-PCR. Leishmania species was identified as L. donovani by sequencing of the amplified products. In order to demonstrate the presence of Leishmania in dogs, another 30 dogs were captured in 2013 from the same sampling area. The microscopic examination of peripheral blood smears was negative for Leishmania infection in all dogs. The rK39 ICT detected one positive dog, which was sacrificed to collect lymph nodes, liver and spleen. Both microscopic analysis and PCR were negative for these tissue samples. Although our findings support the assumption that dogs might be a probable animal reservoir for L. donovani, further studies are required to understand the role of dogs in the transmission VL.
4. Chihiro Kanno (Lab. of Theriogenology, D2)
"Development of the procedure for simultaneous evaluation of multiple characteristics of bovine spermatozoa by flowcytometry"
Introduction
The objective of this study was to develop an objective evaluation procedure to estimate viability, acrosomal integrity, and mitochondrial membrane potential of bull spermatozoa simultaneously by flowcytometry. Firstly, we used frozen-thawed semen mixed with 0%, 25%, 50%, 75% and 100% dead spermatozoa. Semen was stained using three staining solutions: SYBR-14, propidium iodide (PI), and phycoerythrin-conjugated peanut agglutinin (PE-PNA), for the evaluation of spermatozoon viability and acrosomal integrity. Then, the characteristics evaluated by flowcytometry and by fluorescent microscopy were compared. In terms of the results, the characteristics of spermatozoa evaluated by two procedures were similar. Secondly, we attempted to evaluate viability, acrosomal integrity, and also mitochondrial membrane potential of spermatozoa by flowcytometry using conventional staining (SYBR-14, PI, and PE-PNA) combined with MitoTracker Deep Red (MTDR) staining. Then, the spermatozoon characteristics evaluated by flowcytometry using quadruple staining were compared with those of staining using SYBR-14, PI, and PE-PNA and staining using SYBR-14 and MTDR. From the obtained results, there were no significant differences in all characteristics of spermatozoa evaluated by quadruple staining and the other procedures. In conclusion, staining procedure using SYBR-14, PI, PE-PNA, and MTDR for flowcytometry can evaluate viability, acrosomal integrity, and mitochondrial membrane potential of bovine spermatozoa simultaneously.
 |
 |
| ● Date: | November 6 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. May June Thu (Unit of Risk Analysis and Management, CZC, D2)
"Genetic characterization of Rickettsia spp. circulating in Japan"
Introduction
Rickettsiae are obligate intracellular gram-negative bacteria that cause rickettsioses in humans. They are divided into two groups; the spotted fever group (SFG) transmitted by ticks and the typhus group (TG). In Japan, approximately 40 cases of Japanese spotted fever (JSF), caused by Rickettsia japonica, are reported annually. The previous studies reported only partial sequences of rickettsiae circulating in Japan. Therefore, isolation of diverse Rickettsia including non-JSF Rickettsia, followed by genetic characterization at the whole genome level, is required to evaluate the potential risk of the emergence of non-JSF rickettsiosis. From 2013 to 2015, a total of 2,175 ticks, including 17 different tick species, were collected from 13 different prefectures in Japan. DNA samples were examined for Rickettsia infection by real-time PCR. Positive samples were used for genetic characterization by conventional PCR and sequencing. Out of 2,175 ticks, 404 samples (18.6%) were infected with Rickettsia spp. Sequencing analysis of the amplified PCR products indicated the presence of 19 different genotypes. Tick homogenates of Rickettsia-positive samples were inoculated into ISE6 (tick) and C6/36 (mosquito) cell lines. Six different genotypes of Rickettsia spp. were isolated from ISE 6, while one genotype was isolated from C6/36.
2. Ai Dantsuka (Lab. of Anatomy, D1)
"Elucidation of biological characteristics determined by genomic diversity using MRL/MpJ as a mouse model"
Introduction
The mammalian genome contains its own genetic information. Genomic diversity is crucial for the species-specific characteristics in organ morphology and function. Genomic mutations also contribute to their developments, however, they often cause diseases in both humans and animals. Experimental animals, especially inbred mice, are valuable to clarify the relationship between genetic factors and phenotypes because of their abundant biological information.
Inbred MRL/MpJ mice show several unique phenotypes in urogenital organs or immune system caused by the genetic factors in MRL/MpJ-type genome. To discover novel genetic factors making MRL/MpJ unique, I established genetically modified MRL/MpJ, carrying transgenic oncogene or a mutation of muscular dystrophy. Then their phenotypes are being compared to those in other inbred strains.
In this study, I introduce a transgenic MRL/MpJ-Tg(SV40-Ahr)1Poe, expressing the constitutively active aryl hydrocarbon receptor (CA-Ahr) under the control of SV40 promoter. Although original-type (FVB/N) CA-Ahr mice develop stomach tumors in the glandular part, the macroscopic severity of tumors in CA-Ahr mice seemed to be milder in MRL/MpJ-background than in original-type. Therefore, MRL-MpJ mice could be a useful model to elucidate genomic diversity, in particular in the genomic factor-dependent differences of tumorgenesis.
3. Nao Eguchi (Div. of Global Epidemiology, CZC, D2)
"The Mechanism of Inhibition of Influenza Virus Infection by M2 specific Antibody"
Introduction
The antigenic variability of Influenza A virus (IFV) hemagglutinin (HA), which is the principal target of neutralizing antibodies, is a major problem for influenza vaccine manufacturing. In contrast, the M2 protein, one of the envelope proteins of IFVs, is known to have highly conserved amino acid sequences regardless of the HA antigenicity, and expected as a potential target for universal influenza vaccines. It was shown that a monoclonal antibody rM2ss23 recognizing the M2 extracellular domain (M2e) did not neutralize infectivity of an influenza A virus strain, A/Aichi/2/68 (H3N2) (Aichi), but reduced the number and size of plaques of this strain when infected cells were cultured in the presence of the antibody. However, interestingly, rM2ss23 did not show any antiviral effects against another strain, A/Puerto Rico/8/34 (H1N1) (PR8), despite its ability to similarly bind to M2e of PR8. A reverse genetics approach revealed that rM2ss23 reduce the plaque formation of the reassortant viruses containing Aichi HA and M segments but not that of the virus containing M segment alone. For future research, we are planning to elucidate the antiviral mechanism of rM2ss23 in more detail.
4. Kochi Toyomane (Div. of Infection and Immunity, CZC, D2)
"Investigation of the molecular mechanism of AtxA-mediated virulence regulation on Bacillus anthracis"
Introduction
Anthrax is caused by Bacillus anthracis, a Gram-positive, spore-forming bacterium. The pathogenicity of this bacterium depends on two plasmids encoding the most important virulence factors. One encodes anthrax toxin composed of protective antigen, edema factor and lethal factor, while the other encodes capsule synthases. Expression of these factors is coordinately induced at transcription level. This regulation is mediated by a transcription factor of B. anthracis, AtxA. Since it is reported that deletion of atxA leads to low production of virulence factors and attenuation of B. anthracis, AtxA is thought to be a key factor for pathogenicity of anthrax. In spite of its important role in B. anthracis infection, the molecular mechanism by which AtxA controls the expression of virulence factors is not elucidated. To identify and characterize the factors regulated by AtxA directly, we constructed a reporter system. Using the system, we evaluated the interaction between AtxA and a promoter region that is predicted to be binding region of AtxA. As a result, the direct interaction between AtxA and the promoter region was not observed. It is suggested that AtxA controls the transcription of the target genes by unknown mechanisms rather than direct interaction with the promoter regions.
 |
 |
| ● Date: | October 2 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Khoirun Nisa (Lab. of Internal Medicine, D2)
"The Application of Contrast-Enhanced Ultrasonography (CEUS) of Duodenum to Diagnose Chronic Enteropathy in Dogs: Repeatability study"
Introduction
Contrast-enhanced ultrasonography (CEUS) with microbubble as contrast agent can detect pathological changes in intestine by qualitative and quantitative assessment of its perfusion. It may provide a non-invasive modality without radiation that not only support in finding definitive diagnosis but also enable disease monitoring of canine chronic enteropathy (CE). Repeatability of duodenal CEUS is necessary for technique validation and determining reliable measurements to detect changes related to intestinal disorders. Five healthy beagles (age 1-4 year, body weight 9-12 kg) were enrolled for this study. Duodenum was imaged following intravenous injection of Sonazoid® (0.01 ml/kg). Region of interest (ROI) were drawn in duodenal mucosa to create time-intensity curve (TIC). Perfusion parameters including time [wash-in 30% (WI-30), wash-in 70% (WI-70), time-to-peak (TTP), wash-out 70% (WO-70), wash-out 30% (WO-30)] and intensity parameters [peak intensity (PI), area-under-curve (AUC), peak-phase-rate (PPR)] were derived from TIC. CEUS was performed three times within one day (4-hour-interval) and in two different days (1-week-interval) to evaluate intraday and interday variability, respectively. Both time and intensity parameters except WO-30 were relatively stable with intraday and interday coefficient of variation (CV) for most parameters ≤20% (range, 8.30% to 20.31%). Duodenal CEUS could be reliable to evaluate changes related to CE in dogs.
2. Jeewan Thapa (Div. of Bioresources, CZC, D3)
"Molecular characterization of Mycobacterium orygis isolates from animals of South Asia"
Introduction
Mycobacterium orygis, a new member of Mycobacterium tuberculosis complex (MTC) species cause tuberculosis (TB) in humans and animals. We have isolated this bacterium from wild animals in Nepal and captured monkeys and dairy cattle in Bangladesh. MTC isolates from a spotted deer and a blue bull in Nepal and from a dairy cattle herd and captured rhesus monkeys in Bangladesh were included. Most of these animals had TB suspected lung lesions, these lesions were later cultured in LJ media. Spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR), region of difference analysis and SNP detection were performed to ascertain species and molecular epidemiology of the isolates. All the isolates had a typical spoligotype, SIT587 in SpolDB4 database, corresponding to M. orygis and in an agreement with many other reported isolates. RD analysis (cfp32: +, RD9 - and RD12-) and SNP detection of gyrB, mmpL6, TbD1, PPE55 and Rv2042c confirmed the isolates to be M. orygis. The MIRU-VNTR analysis of these isolates grouped them into four MIRU-VNTR types. Previous studies have reported M. orygis from humans in South Asia. These finding suggests its endemic prevalence in the sub-continent. Moreover, genetic diversity of these isolates indicates a long history of prevalence in South Asia. We hope these findings can be helpful to understand the situation of tuberculosis in the region.
3. Eugene Chisela Bwalya (Lab. of Veterinary Surgery, D3)
"Pentosan polysulphate downregulates iNOS, c-JUN and HIF-1α expression in IL-1β- stimulated canine articular chondrocytes"
Introduction
Stimulation with 1, 5, 10 and 20 ng/mL rhIL-1 for 8 hours significantly upregulated (p < 0.05) iNOS, C-JUN, HIF-1α and HIF-2α mRNA expression. Pre-incubation of chondrocytes with 1, 5, 15 and 40 μg/mL PPS before rhIL-1β-stimulation resulted in a dose-dependent downregulation of iNOS, c-JUN, and HIF-1α relative to the positive control. c-JUN and HIF-1α were significantly downregulated (p < 0.05) at all PPS concentrations. Although both HIF-1α and -2α were upregulated by rhIL-1β and significantly associated (p < 0.001) with iNOS expression (R2 = 0.61, correlation coefficient = 0.8 for HIF-1α versus R2 = 0.61, correlation coefficient = 0.61 for HIF-2α), HIF-1α was significantly downregulated at all PPS concentrations suggesting a protective role of PPS from HIF-1α catabolic effects. HIF-1α (and not HIF-2α) may be involved in the induction of iNOS and promotion of cartilage matrix degradation. HIF-2α mRNA remained upregulated at almost all PPS concentrations suggesting a chondroprotective role. The expression of iNOS mRNA in rhIL-1-untreated chondrocytes suggest post-transcriptional regulation. The study has for the first time demonstrated that PPS can ameliorate IL-1β driven OA via effective inhibition of iNOS, c-JUN and HIF-1α mRNA.
4. Heidi Lynn Tessmer (Div. of Bioinformatics, CZC, D3)
"Computational Analyses of Host and Viral Genomes"
Introduction
As our ability to obtain genomic data increases, so does our ability to conduct in-depth analysis of diseases, uncovering previously unknown patterns of evolution and spread. In the future, our ability to effectively combat zoonotic infectious diseases, such as influenza and Ebola virus disease, will require a thorough understanding of the evolutionary mechanisms of the viruses. These mechanisms can most efficiently be discovered through computational analysis of genomes from the nucleotide and protein levels up. This research focuses on rudimentary analysis of single and dinucleotide content in numerous viral and host species. Previously identified trends in C+G content, as well as dinucleotide composition in these viruses, were corroborated and new viruses were analyzed, specifically the 2009 pandemic influenza virus and the Ebola virus from 1976 to the current epidemic. Our findings show similar patterns between human and avian H3N2 viruses, as well as between human influenza and the Ebola virus. Further research is needed to clarify whether these similarities are characteristic of all viruses or specific to the ones which we have chosen.
 |
 |
| ● Date: | September 4 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Nan Aye Thida Oo (Div. of Bioresources, CZC, D1)
"Detection of pncA gene mutations in multidrug-resistant Mycobacterium tuberculosis isolates from Myanmar"
Introduction
Pyrazinamide (PZA) is a first line anti-tuberculosis drug and used for both short course and multi-drug resistant tuberculosis (MDR-TB) treatment regimens. It is a pro-drug and activated by pyrazinamidase which is encoded by pncA. Numerous studies reported that mutations of pncA were responsible for PZA resistance by losing or reducing PZase activity. The aim of present study was to detect the frequencies and patterns of pncA mutation among MDR-TB strains isolated in Myanmar. Clinical MDR-TB isolates were collected and PCR amplification and DNA sequencing of pncA were carried out to detect mutations. Of 369 isolates, 230 samples were processed for DNA sequencing. Mutations in pncA were identified in 96 (41.7%) isolates, while 134 (58.3%) samples had wild type sequences. A total of 69 different types of mutation were dispersed on the pncA and thirty-four of which were found to be novel mutations. This study showed that there were new and diverse mutations on pncA and further studies of correlation between mutations in pncA and alteration of pyrazinamidase activity are needed. Mutation patterns of pncA can be applied for the future development of rapid genotypic PZA susceptibility assays which can lead to great achievement in tuberculosis treatment and control strategies.
2. Yusuke Izumi (Lab. of Advanced Veterinary Medicine, D2)
"Isolation and characterization of tumor endothelial cells"
Introduction
Tumor angiogenesis is the growth of new blood vessels from the existing vasculature toward tumor tissues. Tumor vasculature supplies nutrients and oxygen. It is necessary to tumor growth and metastasis.
Anti-angiogenic therapies function by inhibiting tumor angiogenesis. It causes tumor death indirectly. Several anti-angiogenic therapies have been developed, including bevacizumab, toceranib, and metronomic chemotherapy. Conventional anti-angiogenic drugs target not only tumor endothelial cells (TECs), but also normal endothelial cells (NECs), which may cause adverse effects.
Recently, a few reports indicated that the morphology and genetic expression of human and mouse TECs might differ from NECs. However, there is quite limited information about canine TECs Understanding the characteristics of TECs is essential to the establishment of a new generation of anti-angiogenic therapies with fewer adverse effects. The present study involved the comparison of the characteristics of canine TECs.
In my preliminary experiment, I revealed that canine TECs have different characteristic compared to NECs, such as PD-L1 and CXCR7 expression in the limited histophathological specimens.
In my future study, I would like to do immunohistochemical study using CD105, kind of TEC marker, of various canine tumor tissues and isolate canine TECs using magnetic cell sorting system.
3. Paulina Duhita Anindita (Div. of Molecular Pathobiology, CZC, D2)
"Establishment of compound screening assays for identification of novel antivirals against rabies virus"
Introduction
Rabies is an invariably fatal neurological disease predominantly in rural areas of Asian and African countries caused by rabies virus (RV). Currently available post-exposure prophylaxis procedure and antiviral therapies are not effective for RV-infected individuals after onset of symptoms. Thus, it is urgent to develop new antiviral therapies as an approach to treat exposed and symptomatic individuals. Novel antiviral agents can be identified by screening of compound libraries. As it is known that RV infection does not generate easily discernible cytopathic effects, cell viability assays cannot be employed for such screening of small compounds. To overcome this issue, recombinant RVs (rRVs) encoding NanoLuc luciferase (NanoLuc) inserted in the intergenic region of the RV genome. We determined the growth kinetics of rRVs and demonstrated no decreases in NanoLuc expression over ten passages of virus. We further examined the usefulness of the rRVs for screening of antiviral compounds by treating rRV-inoculated cells with ribavirin, a nucleoside analogue which has a known anti-RV activity in vitro. We demonstrated that NanoLuc signals were decreased in a dose-dependent manner of ribavirin. These results showed that rRVs encoding NanoLuc are a useful tool for screening of antiviral compounds against RV.
4. Yinghua Yang (Lab. of Theriogenology, D2)
"The effect of bone morphogenetic protein 4 (BMP4) on in vitro growth and subsequent development of oocytes derived from bovine early antral follicles"
Introduction
Oocytes in small follicles are less competent than those in large follicles are. The inferior may be caused partly by suboptimal in vitro growth (IVG) culture system in which role of theca cells is absent. Theca cells are considered as an indispensable part to a follicle and one of their major products, BMP4, is an important regulatory factor during follicle development. This study was carried out to test the effects of BMP4 on oocyte IVG and subsequent development. Oocytes from small follicles were treated with three doses (0, 10 and 50 ng/ml) of BMP4 during IVG culture. Throughout IVG, progesterone production from granulosa cell (GC) was suppressed in BMP4 treated groups, consistent with delayed luteinization. After IVG, GC number and oocyte viability were significantly decreased in 50 ng/ml BMP-4 treated group compared to non-treatment group. For subsequent development of IVG oocyte, BMP-4 treated group did not bring improvement in nuclear maturation after in vitro maturation and fertilizability after in vitro fertilization while embryonic development was compromised. In conclusion, BMP4 addition during IVG culture showed the effect of anti-luteinization associated with decreased progesterone level; however, impaired GC proliferation, oocyte survivability and subsequent embryonic development.
 |
 |
| ● Date: | August 14 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Md. Atiqul Islam (Lab. of Experimental Animal Science, D1)
"Analysis for resistance/susceptible genes to Echinococcus multilocularis infection in the mouse"
Introduction
Establishment of congenic lines from B6 and D2 mice using microsettelite PCR could reveal significant resistance/susceptible genetic locus to E. multilocularis infection in the mouse. We already developed 7 congenic and subcongenic lines using microsettelite markers that include chromosomal regions involving the genetic locus responsible for cyst establishment and protoscolex development. For cyst establishment B.D-Emcys1 and D.B-Emcys1 congenic lines and for protoscolex development B.D-Empsc1, D.B-Empsc1, B.D-Empsc1sub1, B.D-Empsc1sub2, and D.B-Empsc1sub1 were established. We challenged these mice with E. multilocularis infection by oral administration of eggs of E. multilocularis. After infection experiments, we found that there was no significant effects of D6Mit188-D6Mit15 locus to cyst establishment. However, we found interesting results in subcongenic line B.D-Empsc1sub1 where 3 mice developed protoscolex among 7 mice. B.D-Empscsub1 subcongenic mice contains D1Mit191-D1Mit14 locus from D2 mice that might possess susceptible gene for protoscolex development. As there were 4 mice in this same line that did not develop protoscolex, so it is required to re-confirm the substituted genomic region. In future identification of the responsible gene and furthermore validation of responsible gene by generating knock-in mice with CRISPR/Cas gene-editing method would be necessary. Our research outcomes would lead novel treatment strategies to control Echinococcosis.
2. Naoya Maekawa (Lab. of Infectious Disease, D2)
"Development of a novel immunotherapy against dog tumors by targeting immunoinhibitory molecules"
Introduction
An immunoinhibitory receptor, PD-1, is expressed on tumor antigen-specific T cells, and suppresses the effector functions of these cells when it binds its ligand, PD-L1. PD-L1 is expressed on many types of tumors and thus plays an important role in the immune evasion of tumor cells. In the field of medical science, therapeutic effect of anti-PD-1 antibody, which inhibits the interaction of PD-1 with PD-L1, has been well characterized and this antibody is expected to be a new treatment for several types of malignancies including advanced melanoma. The purpose of this study is to apply this strategy to dog malignancies.
Immunohistochemical and flow cytometric analyses revealed that dog melanoma, mastocytoma, renal cell carcinoma and other types of tumors express PD-L1. In addition, the expression levels of PD-1 on tumor-infiltrating lymphocytes (TILs) were higher than those of healthy controls. Moreover, dog TILs treated with anti-PD-L1 antibody had improved abilities to produce IFN-gamma, suggesting the potential of this antibody as a novel drug against dog tumors.
Further analysis is required to confirm the therapeutic effect of our antibody on dog patients. After the in vitro analysis, we are planning to perform a clinical trial by using this antibody.
3. Jednipit Borthong (Div. of Bioinformatics, CZC, D2)
"Analysis of bacterial population in water samples collected from the Phadungkrukaseam Canal, Bangkok, Thailand using NGS"
Introduction
Phadungkrukaseam Canal is located in Bangkok, Thailand, is abundant with aquatic plants and animals, and is closely associated with human life. This canal also receives wastewater from household areas and several markets: fresh food, animals, and plants. Thus, this canal may contain a difference of bacterial population. However, this data is still remained and required. In this study, a total of 5 samples (A - E) were analyzed bacterial population based on the nucleotide sequences at the highly variable V3 - V4 region of 16S rRNA using Illumina Miseq Platform. Furthermore, Shannon's index and Simpson's index were used to determine genus richness while SØrensen-Disc coefficient and Kullback-Leibler (KL) divergence were used to compare the similarity of bacterial population. The results revealed that a total of 238 genera (belonging 28 phyla) were found, 27 genera are associated with infectious disease in human. Additionally, alpha diversity indicated that the point C was highly divergent than the other points. SØrensen-Disc coefficient displayed approximately 60 - 70% similarity of bacterial genera detected while KL divergence obviously certified significant difference between bacterial populations. Our results verified that this canal harbors different population of bacteria depending the points and contains some bacteria related with infectious diseases.
4. Minato Hirano (Lab. of Public Health, D2)
"Study on genomic transport-mechanism of the neuronal infection of flaviviruses"
Introduction
Neurological diseases caused by encephalitic flaviviruses are severe and are associated with high levels of mortality. However, detailed mechanisms of the viral replication and pathogenesis in the brain remain poorly understood. Previously, we reported that the replication of tick-borne encephalitis virus (TBEV) altered membrane structure in dendrites of mouse primary neuronal culture, which might be involvegd in the neuro-pathogenicity of TBEV. In the current study, I analyzed detailed mechanisms of the transport of the viral genome to the neurites. Transfection of the DNA-based TBEV replicon caused the viral antigen and genomic RNA accumulations in neurites of the PC12 cells. The viral genomic RNA was still observed in the cells transfected with a replicon not expressing the viral proteins. As well as in the cells transfected with a replicon in which most of the coding sequence was deleted. These results showed that the viral genomic RNA was directly transported to the neurites, and the transport was independent of the viral proteins. The RNA sequence of the 5'-UTR and/or 3'-UTR were required for the transport to the neurites. I will further examine the viral and host factors involved in the transport to reveal molecular mechanisms of TBEV infection in neuronal cells.
 |
 |
| ● Date: | July 10 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Masahiro Sato (Div. of Global Epidemiology, CZC, D1)
"Studies on the biological property of bat-derived influenza viruses"
Introduction
Influenza A virus (IAV) is a member of the family Orthomyxoviridae and have 8 segmented single-stranded negative-sense RNA genomes. IAV is one of the most important zoonotic pathogens. Recently, IAV genomes of novel subtypes (i.e., H17N10 and H18N11) were detected from frugivorous bats in Guatemala and Peru. However, since infectious virus strains have never been isolated, the biological properties of these bat-derived IAVs are largely unknown. In this study, we aim to elucidate receptor molecules required for cellular entry of bat-derived IAVs, and also attempt to obtain information on the host range and zoonotic potential of the virus. First, we will try to rescue a reassortant virus which has HA and NA genes of bat-derived IAV and the other genes of a well-characterized IAV strain (PR8) and use the virus for screening receptor molecules. We will further attempt to generate bat-derived IAVs with a complete set of the 8 gene segments to infect experimental animals. Second, receptor molecules of bat-derived IAVs will be identified by an expression cloning method using cDNA library from IndFSPT1 cells which are potentially susceptible to bat-derived IAVs. Third, we will generate monoclonal antibodies against bat-derived IAVs and identify neutralizing epitopes on HAs.
2. Yuri Sakai (Lab. of Radiation Biology, D2)
"The response of cancer-associated fibroblasts to ionizing radiation and its effect on cancer pathophysiology"
Introduction
To improve the efficacy of cancer radiotherapy, the effect of radiation on cancer cells in tumor has been studied for a long time. On the other hand, the growing evidence suggests that cancer prognosis after treatment is greatly influenced by tumor stroma. Cancer-associated fibroblasts (CAFs) are major components of tumor stroma and secrete various soluble factors, resulting in malignant progression and metastasis. However, there is little information about how CAFs respond to ionizing radiation. Therefore, the aim of this study is to reveal the effect of radiation on the secretory phenotype of CAFs and its effect on cancer pathophysiology. In this study, following studies are planned: (1) production of experimental CAFs (eCAFs), (2) analyses of the soluble factor and gene expression of irradiated eCAFs, (3) analyses of the effect of irradiated eCAFs on cancer cell phenotypes, and (4) analyses of the effect of irradiated eCAFs on cancer pathophysiology. For these experiments, cell cultures and materials for these studies were prepared and some preliminary experiments were performed. The tumorigenicity of BALB/c mouse-derived mammary carcinoma cell line EMT6 was confirmed by inoculating them into mammary fat pads of BALB/c mice, and the metastasis to other organs was investigated. As a result, they formed multiple metastatic lesions in the lung.
3. Chihiro Ishii (Lab. of Toxicology, D2)
"Lead (Pb) poisoning is still a serious problem in raptors from Japan"
Introduction
Lead (Pb) poisoning occurs widely among raptors. Raptors ingest Pb rifle bullet or shot fragments when they consume mammals or birds that were hunted with Pb ammunition. In Hokkaido, the northern island of Japan, the use of Pb rifle bullets for deer hunting has been prohibited since 2001. Incidentally, other areas of Japan, such as Honshu or Shikoku have few regulations and the current studies of Pb poisoning are little clarified. In this study, we measured the Pb concentrations and Pb stable isotope ratios (Pb-IRs) to identify the source of poisoning in 16 species of 174 raptors from Japan. Pb distribution in bones and bone marrows was also measured. This study clarified that accumulation of one third sea eagle which died in Hokkaido exceeded the level of Pb poisoning after the regulation. In Honshu and Shikoku, 5 eagles include endangered species accumulated high Pb concentration, indicating that they were exposed to Pb. Pb-IRs indicate that both of Pb bullets and shot have the risk of causing Pb poisoning in all parts of Japan. Furthermore, there is a possibility that analysis of bones and bone marrows would be effective to reveal Pb exposure and periods of after exposure.
4. Yogendra Shah (Div. of Bioresources, CZC, D2)
"Diversity of MIRU-VNTR among Mycobacterium tuberculosis Central Asian strain family isolates from Nepalese patients"
Introduction
Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) poses major challenges and public health problems in Nepal. Central Asian Strain (CAS) family has been reported as one of the most predominant genotypes of MTB in South Asian countries including Nepal. The main objective of this study was to understand molecular epidemiological feature of MTB CAS Delhi strain circulating in Nepalese patients and to assess its transmission dynamics. A total of 145 MTB CAS isolates from Nepalese patients were analyzed by spoligotyping and 24 loci MIRU-VNTR. Using HGDI, three loci (QUB26, MIRU10, VNTR 424) were found to be "highly discriminatory" (DI>0.6), eleven loci (MIRU26, MIRU31, MIRU40, VNTR1955, VNTR 3960, ETR-A, VNTR 2401, QUB 4156, MIRU39, ETR-F and QUB 11a) were "moderately discriminatory" (DI:0.3-0.6) and ten loci (MIRU16, ETR-C, QUB11b, MIRU4, MIRU2, MIRU20, MIRU23, MIRU 24, MIRU27 and ETR-B) were poorly discriminatory (DI<0.3). By combining both MIRU-VNTR and Spoligotyping comprised eighteen clusters having the 47 isolates of identical pattern where as clustering rate was found to be 20%. The proposed 15 loci MIRU-VNTR typing scheme is well suited to assess the MTB population structure and diversity, trace back the transmission dynamics and epidemiological link among MTB CAS family strain within Nepal and surrounding countries.
 |
 |
| ● Date: | June 12 (Fri), 2015, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Woongchul Shin (Lab. of Biochemistry, DC1)
"Effects of dietary component on the activation, proliferation, and recruitment of thermogenic cells"
Introduction
There are two kinds of fat tissue in mammals, brown adipose tissue (BAT) for heat generation and white adipose tissue(WAT) for energy storage. BAT exclusively expresses uncoupling protein 1 (UCP1), which is responsible for thermogenesis. Interestingly, in response to sympathetic nerve stimulation, WAT changes its metabolic phenotype to BAT-like one (WAT browning); UCP1-expressing brown-like adipocytes, termed as beige adipocyte are induced within the tissue. In the previous study, we found that ginger feeding suppressed diet-induced obesity in mice and that it activate brown adipocyte and also induced beige adipocyte. However, actual contribution of BAT activation and WAT browning to the anti-obesity effect by ginger is not clear. Thus, our purpose of study is to evaluate how brown and beige adipocytes contribute to the anti-obesity effects of ginger by using UCP1-KO mice. Also, although recent studies revealed that beige adipocyte arises from a progenitor cell different from brown adipocyte, the cellular origin of the progenitor is unknown. WAT is shown to contain mesenchymal stem cells, almost identical to those from bone marrow (BM). Also, it has reported that BM-derived stem cells migrate to WAT following to some pharmacological stimulation. Thus, we will examine whether BM-derived residential stem cells differentiate into beige adipocytes.
2. Teppei Ikeda (Lab. of Anatomy, D2)
"Studies on mechanism of skeletal muscle remodeling - Analysis of the molecules contributing to myoblast differentiation and growth -"
Introduction
All animals repeat disruption and regeneration of myofibers, that is, skeletal muscle remodeling. Aging or several diseases could cause disorder of skeletal muscle remodeling, which leads to breakdown and atrophy of skeletal muscle, resulting in astasia. These pathological progressions can be serious problems in livestock animals and elder companion animals since they get in difficulty in feeding, which leads to euthanasia and continuing care. Muscle repair is a multistep process that includes myofiber degeneration and regeneration. After damage to the myofibers, satellite cells activate, proliferate, and give rise to a population of transient-amplifying myogenic cells, called myoblasts, which express myogenic regulatory factors (MRFs) such as myogenic differentiation 1 and myogenic factor 5. Myoblasts subsequently express another MRF called myogenin, commit to terminal differentiation, and fuse to reconstruct their host myofibers or to generate new myofibers and repair the damaged tissue. Although much is known about the transcription factors and signaling pathways involved in the skeletal muscle remodeling, the detailed process is still incompletely understood. In this study, we widely investigated the morphological and pathological features of skeletal muscles in C57BL/10ScSn-Dmdmdx (mdx) mice, known as a representative model for Duchenne muscular dystrophy in humans.
3. Asami Nishimori (Lab. of Infectious Disease, D2)
"Large-scale production of the monoclonal antibody against bovine immunoinhibitory molecules for clinical application in cattle"
Introduction
In previous study, we revealed one of the disease developing mechanisms in bovine leukemia virus (BLV) infection. Programmed death-1 (PD-1) is a receptor on T cells, and the interaction of PD-1 and its ligand (programmed death-ligand 1: PD-L1) inhibits T-cell activation. Our previous reports showed that the both of PD-1 and PD-L1 expression were upregulated in line with disease progression of BLV infection, and moreover, the blockade of the PD-1/PD-L1 interaction by using antibodies increased IFN-γ production in vitro. These findings suggest that the PD-1/PD-L1 blockade could be clinically applied as a novel therapeutic method for controlling BLV infection. The next stage of our research is to evaluate the in vivo function of the blocking antibody. However, there are several problems to apply this antibody in cattle; 1) because the blocking antibody is delivered from rats, it might induce severe inflammation if frequently administered; 2) a large amount of antibody is required for clinical trial in cattle. Thus, we are planning to develop rat-bovine chimeric antibody and to establish cell clones for large-scale antibody production. Now, cell groups we established were producing 21.12mg/L of the chimeric antibody, and further steps to obtain more superior cell clones is ongoing.
4. Yuji Wada (Div. of Molecular Pathobiology, CZC, D2)
"Development of novel chemical therapy for Chikungunya fever"
Introduction
Chikungunya fever (CHIKF) is caused by Chikungunya virus (CHIKV) belonging to genus Alphavirus, family of Togaviridae. The main symptom of CHIKF is arthritis which often persists several months to over year, and sometime fatal for children, elderly and the persons who have underlying condition. CHIKF has now widely spread and caused severe damage for health and economic in the world because of no effective treatments and approval drugs. In order to control CHIKF, it is necessary to develop therapeutic strategies. First, we conducted the screening of chemical compounds library to find candidate compounds which inhibit CHIKV infection. As a result, we found the some candidate compounds. Additionally, we confirmed one of the candidate compounds inhibits CHIKV replication significantly. To unveil the CHIKV inhibitory mechanism of these compounds, we performed selective amplification of CHIKVs (res-CHIKV) whish are resistant to the inhibitory effect of candidate compounds. Subsequently, we performed full genome sequence analysis to find characteristic amino acid change in res-CHIKVs. As a result, we found three or four characteristic amino acid change from two clones of res-CHIKV. Now, we assess the relation between each mutation and resistant ability against candidate compounds by reverse engineering.
 |
 |
| ● Date: | May 15 (Fri), 2015, 18:00 - 19:55 |
| ● Venue: | Lecture Hall |
1. Reiko Akamatsu (Div. of Infection and Immunity, CZC, DC2)
"Multilocus sequence typing of Bacillus cereus nosocomial isolates"
Introduction
Bacillus cereus is associated with food-borne illness with vomiting and diarrhea. It is known that several strains among B. cereus potentially cause severe extra-intestinal infections. Pathogenic B. cereus caused nosocomial outbreaks and were lethal to patients in Japan. Based on multilocus sequence typing (MLST) and phylogenetic analysis, pathogenic B. cereus, which were isolated in Gifu University hospital and Jichi Medical University (GTC strains), are closely related to B. anthracis than B. cereus standard strain. However it is difficult to discriminate pathogenic B. cereus strains from less virulent or non-pathogenic strains. In 2013, National Cancer Center (NCC) had an outbreak of B. cereus infection. Twenty-nine strains of B. cereus were isolated from human blood samples and hospital appurtenances. From the result of MLST, we found that the properties of 8 strains were similar to GTC strains. There is a possibility that NCC outbreak was caused by B. cereus strains which were similar in nature to GTC strains.
2. Chiho Kaneko (Div. of Risk Analysis and Management, CZC, D2)
"Studies for enhancement of strategies for the control of rabies in Zambia"
Introduction
Rabies still causes 55,000 deaths per annum, predominantly among children in Asia and Africa. The execution of control of dogs undertakes the success of control of rabies, yet the situations in the endemic countries are confronted with various challenges. This research aims to understand the current status of rabies and evaluate the current control measures against rabies in Zambia. It will be succeeded by the risk analysis of rabies in Zambia using relevant data. The comprehensive outcome will apply to establish the future strategy for the control of rabies. The field survey collecting canine blood samples was conducted to evaluate the immunization coverage against rabies in domestic dogs and to evaluate the efficacy of domestic produced rabies vaccine. For the collection of canine blood samples, the mass vaccination campaigns were held in the target local districts. 736 dogs were vaccinated and then 265 blood samples were collected in total. In Lusaka, the capital of Zambia, two-stage cluster sampling was carried out. 278 canine blood samples were collected through the programme. KAP (Knowledge, Attitude and Practice) survey for the dogs' owners was also conducted and 224 responses were obtained. Subsequent analysis is in progress.
3. Suppalak Kaewkwan (Lab. of Wildlife and Medicine, D2)
"Genetic status monitoring of wild Asiatic black bears in Thailand forests"
Introduction
The Ursidae family is the largest carnivore group that has many published of brown bears and American black bears from Europe and North America. On the other hand, the status of wild Asiatic black bears in Thailand is little known, especially the genetic variation of this species has never been evaluated. Thai researchers are interested in wild bear ecology, habitat, food variety, and bear activity patterns. My study plan is focusing on genetic variation of wild Asiatic black bear in the forest complexes that are continuous evergreen forest in northern, northeastern and western Thailand. Hair samples will obtain from a rub tree trapping method using a DNA hair sampling technique. A set of hair trapping contained 20 coiled wires on the tree (5 rows and 4 columns) and a camera trap was set up at the opposite side. Wire has 1 inch with 0.5 inch spike at the end. All designed traps will be distributed in 5 wildlife sanctuaries with 6x6 km. (36 sq.km.) grid cell size that referred minimal Asiatic bear home range. We will put at least 10 traps near watershed areas where bears are commonly found.
4. Manyando Simbotwe (Div.of Infection and Immunity, CZC, D2)
"Development of an enzyme-linked immunosorbent assay for the sero surveillance of anthrax in endemic areas"
Introduction
Bacillus anthracis, protective antigen (PA) is composed of four structural domains and is responsible for the shuttling of edema factor and lethal factor into the host cell during the pathogenesis of anthrax. Each of the four domains of PA plays a role in the host intoxication process. Current serological assays including ELISAs have been developed based on PA, to analyze immune responses against B. anthracis infection. However, one of their major limitations is their inability to distinguish vaccine-induced immunity from natural infection. In order to effectively carry out sero surveillance and evaluate the immunization status of cattle in anthrax-endemic areas, the development of an assay that will overcome the above mentioned limitation is necessary. In this study, to prepare antigens for the new ELISA, we cloned and expressed each of the four PA domains individually. The recombinant proteins PA-D1, PA-D2, PA-D3 and PA-D4 were expressed as GST fusion proteins in Escherichia coli. The results of the protein expression revealed that PA-D1 was expressed in the soluble fraction of the cell lysate. On the other hand, PA-D2, PA-D3 and PA-D4 had low levels of expression in the soluble fraction. Predominant expression of the proteins was in the pellet fraction as inclusion bodies.
5. Motofumi Suzuki (Lab. of Radiation Biology, DC3)
"MPS1 inhibition enhances chemosensitivity to etoposide and paclitaxel but not radiosensitivity in murine SCCVII cells"
Introduction
In eukaryotes, the spindle assembly checkpoint (SAC) is an mechanism to ensure the proper chromosome segregation. Recently, several studies have demonstrated that MPS1 plays an important role in SAC signaling. Furthermore, MPS1 inhibitors have been shown to exhibit potent cytocidal effect, implying the potential of MPS1 as a novel target of cancer treatment. However, it remains to be determined how MPS1 inhibition influences the cell-killing effect by ionizing radiation or anti-cancer drugs. Therefore, our aim in this study is to elucidate the relationship between MPS1 and cellular radio-/chemo-sensitivity. To evaluate whether MPS1 inhibition influences cellular radio/chemosensitivity, we analyzed cellular survival after exposure to cytotoxic treatments with or without MPS1 inhibitors by clonogenic survival assay. Whereas both MPS1 inhibitors sensitized SCCVII cells to anticancer drugs, they did not enhance cellular radiosensitivity. Furthermore, to evaluate the effect of MPS1 inhibitors on cell cycle progression, we assessed cell cycle distribution after cytotoxic treatments with or without MPS1 inhibitors by flow cytometry. We found that tetraploid cells were increased when cells were co-treated with MPS1 inhibitors and anticancer drugs, while MPS1 inhibition had little effect on cell cycle status after irradiation. This study revealed that MPS1 is a potential target to improve cancer chemotherapy.
 |
 |
| ● Date: | April 17 (Fri), 2015, 18:00 - 20:00 |
| ● Venue: | Lecture Hall |
1. Wallaya Phongphaew (Div. of Molecular Pathobiology, CZC, DC2)
"Role of the Endoplasmic Reticulum-associated protein degradation (ERAD) in West Nile virus infection"
Introduction
ERAD is protein quality control of cells. It is related to degradation of misfolded-proteins to prevent cell death caused by accumulation of abnormal proteins. The strategies can be divided into four major steps including misfolded-protein recognition, retranslocation the misfolded-proteins into cytosol, ubiquitination to target for proteasome and degradation by proteasome. However, ERAD is not only protein quality control, but it also plays some roles in infection of many viruses. Human immunodeficiency virus type 1 (HIV-1) and herpesviruses manipulated ERAD to evade from host immune response. While hepatitis c virus (HCV) was increased when ERAD or ERAD components were inhibited. HCV envelope protein was target for ERAD. Decrease in ERAD activity could retain HCV envelope protein in the infected cells and increase production titer of infectious particles. On the other hand, the present results indicate that inhibition of ERAD using chemical compound, Eeyarestatin 1 decrease WNV infection. This compound target SEC61 and Valosin containing protein, the components in ERAD machinery. However, the mechanism remains unknown and requires further study to elucidate. We have planned to use other ERAD inhibitor and ERAD gene silencing to confirm whether ERAD inhibition effect to WNV infection and reveal that which steps of WNV replication is involved in ERAD machinery.
2. Yuji Hirai (Lab. of Veterinary Hygiene, D2)
"Analyses of cell-attachment and growth of Listeria monocytogenes in intestinal models for identification of novel virulence-associated markers"
Introduction
Listeria monocytogenes (Lm) causes severe and fatal foodborne diseases, such as gastroenteritis, meningoencephalitis, and maternofetal infections. Lm is isolated from patients, food products, and the environment. There are several genetic analyses for estimating the virulence of Lm isolates, however, genetic analyses are still short of the estimation of pathogenicity of Lm isolates. Thus, further virulence-associated markers are required to estimate the pathogenicity of isolates. In Lm infections, Lm multiplies in the intestinal lumen, cross the intestinal barrier and disseminate to the mesenteric lymph nodes, spleen, and liver upon ingestion of contaminated food. Cytotoxicity of Lm to intestine cells may provide the information for the assessment to the pathogenicity of Lm. Thus, we attempted to evaluate interactions of Lm with intestinal cells on cell-attachment, intracellular growth, and cell-to-cell spread. In this year, we carried out biofilm forming assay, cell-attachment assay, and plaque forming assay. We tentatively selected 20 representative isolates, including 9 clinical isolates and 11 food isolates, from 171 isolates based on the results of biofilm assay, and used them for following analyses. No difference was observed in results of clinical or food isolates in cell attachment assay and plaque forming assay. Hierarchical cluster analysis revealed that the 20 representative isolates were successfully classified into three clusters based on results of three in vitro assays. In the future, fine cluster analysis will be carried out using a number of Lm isolates and additional parameters, including molecular epidemiological characters. Pathogenicity and whole genome sequences of the representative isolates of each cluster will be analyzed for the identification of novel virulence markers.
3. Lesa Angela Thompson (Lab. of Toxicology, D2)
"Chicken - source of nutrition or toxin?"
Introduction
WHO estimated that in 2013 there were 198 million cases of malaria, with 584,000 deaths. 90% of these deaths occur in Africa. The disease is caused by species of Plasmodium parasite and spread by Anopheles mosquitoes. A common method of control is to use pesticides such as dichloro-diphenyl-trichloroethane (DDT) to reduce numbers of the vector. However, this chemical has toxic effects in many species - problems such as population declines in wild avian species resulted in the banning of DDT in many countries and its use is now strictly regulated. Although thought to be relatively safe for humans, reports have shown bioaccumulation to occur and suggested exposure may result in neurotoxic, carcinogenic, immunotoxic and reproductive effects. Muscle samples from free-ranging chickens (n=48) in the KwaZulu-Natal region of South Africa, where DDT is sprayed annually in homes were analysed for DDT content. The predominant DDT congener detected was the p,p'-DDE metabolite (mean, 10.09ug/g wet weight). The mean ΣDDT detected in free-ranging chicken muscle was 11.98ug/g wet weight. However, most chicken eaten in the area is intensively-reared broilers purchased locally. Analysis of such broilers (n=6) showed much lower levels of ΣDDTs (mean, 0.09ug/g wet weight).
4. Memi Muto (Lab. of Public Health, D2)
"Isolation and characterization of tick-borne encephalitis virus from Ixodes persulcatus in Mongolia in 2012"
Introduction
Tick-borne encephalitis virus (TBEV) is a zoonotic virus belonging to the genus Flavivirus, in the family Flaviviridae. The virus, which is endemic in Europe and northern parts of Asia, causes severe encephalitis. Tick-borne encephalitis (TBE) has been reported in Mongolia since the 1980s, but details about the biological characteristics of the endemic virus are lacking. In this study, 680 ticks (Ixodes persulcatus) were collected in Selenge aimag, northern Mongolia, in 2012. Nine Mongolian TBEV strains were isolated from tick homogenates. A sequence analysis of the envelope protein gene revealed that all isolates belonged to the Siberian subtype of TBEV. Two strains showed similar growth properties in cultured cells, but their virulence in mice differed. Whole genome sequencing revealed only thirteen amino acid differences between these Mongolian TBEV strains. Our results suggest that these naturally occurring amino acid mutations affected the pathogenicity of Mongolian TBEV. Our results may be an important platform for monitoring TBEV to evaluate the epidemiological risk in TBE endemic areas of Mongolia.
5. Tomoyuki Yamaguchi (Div. of Bioresources, CZC, DC3)
"The action of DNA gyrase and quinolone resistance"
Introduction
DNA gyrase is an essential enzyme for DNA replication and transcription of bacteria. It catalyzes the negative supercoiling of the bacterial circular DNA by cleaving the double strands, passing another part of DNA through the cleavage and resealing the DNA. Quinolone, a family of antibacterial drugs, interferes with the DNA-resealing of DNA gyrase and induce malfunction of DNA replication or transcription. Amino acid substitution in the quinolone binding site of DNA gyrase may confer quinolone resistance on bacteria. In Mycobacterium leprae that is the cause of leprosy, amino acid substitution at position 89, 91 and 95 on DNA gyrase subunit A can confer quinolone resistance. In our recent study, we conducted in vitro assays with moxifloxacin (MXF), sitafloxacin (SIT) and a newly developed quinolone to find out effective quinolones even against resistant strains. Their inhibitory activities on recombinant M. leprae DNA gyrases with four types of GyrA (wild type, Gly89Cys, Ala91Val and Asp95Gly) were examined and compared. We found SIT and the newly developed quinolone have much higher efficacy as DNA gyrase inhibitors than MXF.
 |
 |
| ● Date: | March 13 (Fri), 2015, 18:00 - 19:55 |
| ● Venue: | Lecture Hall |
1. Tapiwa Lundu (Lab. of Public Health, DC1)
"Studies on the ecology of Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV) and the interactions of viral proteins in infected cells"
Introduction
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonosis in China, Japan and South Korea, caused by SFTS virus (SFTSV), a novel phlebovirus in the family Bunyaviridae. The disease is characterized by fever, thrombocytopenia, coagulopathy and a case fatality rate of 2 to 15%. The SFTSV genome consists of L, M and S segments which encode viral RNA polymerase, glycoproteins (Gn and Gc), nucleoprotein (NP) and a nonstructural (NSs) protein, respectively. Haemaphysalis longicornis, Rhipicephalus microplus and Amblyomma testudinarium ticks are the vectors for SFTSV. Several mammals are potential reservoir hosts of SFTSV because anti-SFTSV antibodies have been detected in both wild and domestic animals. To determine the sero-prevalence of SFTSV antibodies in rodents, we used ELISA and IFA to test 1783 rodent sera collected between 1997 and 2012 in western Japan, Russia and Mexico. No antibodies were detected in rodent sera from the three countries. The result suggests a very low SFTSV prevalence in rodents during the survey period. It is suggested that NP and NSs interact with each other and are involved in virus replication. To understand the mechanism of this interaction in transfected and SFTSV infected mammalian cells, plasmids expressing NP and NSs protein were constructed.
2. Tomoya Morita (Lab. of Internal Medicine, D1)
"Establishment of a new right heart function test by echocardiography in dogs"
Introduction
Heart disease is important for dogs. Until now, heart function test has been focused on left heart, and right heart function has been underestimated. Right heart dysfunction has recently been shown to predict poor prognosis and symptom. Therefore, right heart function test attracts attention in human medicine. Echocardiography is the mainstay of evaluation of heart function because it is noninvasive and repeatable test. However, the quantitative assessment of right heart function using conventional echocardiography is challenging, because of the complex shape of right ventricular (RV). Recently, right heart function can be more easily measured thanks to a new echocardiography technique, called speckle tracking echocardiography (STE). STE enable to quantify right ventricular systolic performance, and this value (RV strain) has been shown to predict prognosis, severity of the patients with several heart disease. On the other hand, STE has not applied to dogs so far. Therefore, we assess the repeatability and effect of hemodynamic change on RV strain. Good repeatability in normal beagle dogs was shown in this study. RV strain decreased in acute pressure overload condition caused by infusion U46619, which is pulmonary artery vasoconstrictor. For the future, we`ll create the chronic pressure overload model dogs and measure RV strain.
3. Nyamsuren Ochirkhuu (Laboratory of Infectious Diseases, D2)
"Study on epidemiological and immunological research for intractable infectious diseases in livestock"
Introduction
Out of 339 samples collected from cattle in the Luzon island of the Philippines, 324 (95.5%), 154 (45.4%), 209 (61.6%), 140 (41.3%), and 2 (0.6%) were positive for Anaplasma marginale, Babesia bigemina, Babesia bovis, Theileria spp., and Trypanosoma evansi infections, respectively. Phylogenetic analyses showed that A. marginale 16S rRNA gene detected in this study was in a same lineage across the world, most of B. bovis RAP-1 and B. bigemina AMA-1 genes detected were in same and similar lineage from Asian isolates, and the MPSP genes of Theileria spp. belonged to the group of T. sergenti and T. buffeli.
The seroprevalence of Mycobacterium avium subspecies paratuberculosis in Mongolian cattle was examined by ELISA, and 3 (0.84%) seropositives were detected from 356 cattle. The bovine leukemia virus genome was detected by a nested PCR, and 20 (3.86%) were positive out of 517 samples. Phylogenetic analysis showed that 6 isolates were classified into genotype-4, one into genotype-7 and the other 4 into genotype-1. Bovine viral diarrhea virus was detected by immunochromatographic tests, and 6 (5.5%) were positive out of 110 serum samples. However, the virus genome was not detected by RT-PCR in the cattle positive for the BVDV antigen.
4. Wakako Furuyama (Div. of Global Epidemiology, CZC, D2)
"Host signaling pathways involved in antibody-dependent enhancement of Ebola virus infection"
Introduction
Antibody-dependent enhancement (ADE) of Ebola virus (EBOV) infection has been shown to depend on the cross-linking of virus-antibody or virus-antibody-complement complexes to cell surface Fc or complement receptors, leading to enhanced infection of susceptible cells. In this study, we examined whether the Fc receptor-dependent ADE entry of EBOV requires host cell signaling. ADE-dependent infectivity of vesicular stomatitis virus pseudotyped with EBOV glycoprotein (VSV-EBOV GP) was first compared using Jurkat cells transduced with cDNA encoding full-length Fc gamma receptor IIa (FcγRIIa) or mutant FcγRIIa lacking its cytoplasmic tail (FcγRIIa (CT-)). We found that infectivity of VSV-EBOV GP was significantly enhanced in cells expressing full-length FcγRIIa in the presence of an ADE antibody, whereas the enhancement of infectivity was limited in FcγRIIa (CT-)-expressing cells. To investigate possible signaling pathways downstream of FcγRIIa in the ADE entry of EBOV, ADE-dependent and -independent infectivities of VSV-EBOV GP were compared using K562 cells naturally expressing FcγRIIa in the presence of several inhibitors of FcγRIIa-mediated signaling pathways. We found that the ADE-dependent infection was exclusively reduced by R788 and PP2, inhibitors of Syk or Src family PTKs. These results suggest that the FcγRIIa-mediated signaling pathways are important for ADE of Ebola virus infection.
 |
 |
| ● Date: | February 6 (Fri), 2015, 18:00 - 20:20 |
| ● Venue: | Lecture Hall |
1. Huy Duc Chu (Lab. of Microbiology, DC2)
"The evaluation of hygiene status at live bird markets (LBMs) in Vietnam by virological survey and knowledge, attitude and practice (KAP) survey"
Introduction
The findings of previous study suggested that LBMs have a potential to act more than backyard farms as the primary source of AI virus as well as H5N1 HPAI virus to amplify, maintain, circulate and transmit the virus in the environment in Vietnam (Okamatsu et al., Virus Genes. 2013). It was reported that closing of LBMs was effective solution to minimize a risk of virus transmission to humans. However, closing of LBMs is a difficult challenge at the moment but it can do step-by-step. Because of cultural issues and tradition of live poultry consumption may not be changed immediately. It is important to improve biosecurity of LBMs to minimize a virus transmission to other animals and humans for the control of AI not only in Vietnam but also in Asian countries. We are conducting the virological survey and KAP survey e for the improvement of hygiene status of LBMs for the control of HPAIV. The preliminary results indicated that the AI prevalence at LBMs is quite high, 42.1%. There are 64 AI viruses were isolated including HPAIV H5N1, H5N6 strains.
2. Jemima Tiwaa Marfo (Lab. of Toxicology, D2)
"Determination of urinary neonicotinoid and its relation to some typical symptoms"
Introduction
Neonicotinoid insecticides are used worldwide in agriculture, domestic and public pest control. The Environmental health effects of six neonicotinoids and a metabolite, namely, imidacloprid, acetamiprid, nitenpyram, thiacloprid, clothianidin, thiamethoxam and N-desmethyl-acetamiprid were studied on humans. The association between urinary chlorinated-NN/metabolite and clinical symptoms in 85 subjects from the Japanese general population was evaluated by a prevalence case-control study. The detection of thiamethoxam or N-desmethyl-acetamiprid was correlated to the simultaneous exhibition of postural finger tremor, recent memory loss, headache, general fatigue, palpitation/chest pain, abdominal pain, and muscle pain/weakness/spasm with electrocardiographic findings. Their symptoms mostly continued for several days to months after prohibiting consumption of locally grown produce. The results suggest environmentally exposed NN/metabolite may cause adverse health effects on humans. As a result of the outcome of that study, quantification method for dinotefuran which is mainly used in Japan is currently under investigation. Enzymatic kinetics of neonicotinoids can also provide useful information for detoxification. Therefore, in vitro enzymatic metabolic activity measurement of some of the neonicotinoids such acetamiprid is being studied. Also, methods for other unique metabolites of neonicotinoids such as 6-chloronicotinic acid (common metabolite for imidacloprid, acetamiprid and nitenpyram) and chlorothiazolecarboxaldehyde (common metabolite for clothianidin, thiamethoxam and thiacloprid) in urine shall be developed.
-Introduction of their works by Nagasaki Univ. Graduate Students-
1. Nut Koonrungsesomboon (Institute of Tropical Medicine, Nagasaki University)"Improvement of Layperson's Understanding in Clinical Trial Using SIDCER Methodology: A Randomized-Controlled Study "
Introduction
Introduction: Informed consent form (ICF) is a required document in clinical studies. However, the quality of ICF is problematic due mainly to its extensive length with jargons and a lack of essential elements required by regulations. As such, the Strategic Initiative for Developing Capacity in Ethical Review (SIDCER) has recently proposed the SIDCER methodology for ICF development.
Objective: To evaluate the comprehension of laypersons with two interventions: SIDCER ICF vs. conventional ICF, using a post-test questionnaire as an assessment tool.
Methods: 550 laypersons in Chiang Mai, Thailand, were randomly assigned to read either SIDCER ICF (group I) or conventional ICF (group II) (1:1) and subsequently performed the post-test. The primary endpoint was the proportion of participants who have the post-test score of .80%; the secondary endpoint was the total score of the post-test.
Results: The primary endpoint was higher in group I, compared to group II (82.2% vs. 60.4%, p <0.001). The median total score in group I was higher than that in group II (19/21 vs. 18/21, p <0.001).
Conclusion: The ICF developed based on the SIDCER methodology can enhance the understanding of laypersons in this study. Evaluation of its effectiveness in different types of studies and languages is necessary.
2. Nozomi Imanishi (Nagasaki University Graduate School of Biomedical Sciences)
"Morphological and phylogenetic studies on Anopheline mosquitoes in Hokkaido"
Introduction
There was a survey to refer to this issue previously in 1968-1974, however, no definitive surveys had been carried out thereafter to approach to it systematically. In Kushiro wetland, in northern Japan, there is a newly discovered population of anopheline mosquitoes (=An. sp. Kushiro) that has a unique DNA sequence, different from any of the Anopheles species in Japan. Internal Transcribed Spacer 2 (ITS2) sequences of rDNA identified this population as Anopheles belenrae Rueda, first described in Korea in 2005. However, the taxonomic status of An. sp. Kushiro is still unclear. In this study, further phylogenetic study was performed to ask if An. sp. Kushiro is genetically identical to An. belenrae not only ITS2 region but also DNA barcoding region (partial mtDNA COI). Moreover, further morphological comparison between An. sp. Kushiro and the Korean An. belenrae was performed using paratype specimens.
3. Satoshi Shimada (Institute of Tropical Medicine, Nagasaki University)
"Therapeutic effect of post-exposure treatment with antiserum on severe fever with thrombocytopenia syndrome in a mouse model of SFTS virus infection"
Introduction
Severe fever with thrombocytopenia syndrome is an emerging viral disease that is endemic in China, Korea and Japan. No effective vaccine or specific treatment for SFTS is currently available. In this study, we examined the effects of ribavirin, site-1 protease inhibitor PF-429242, steroids, minocycline and ciprofloxacin, and antiserum from a recovered SFTS case on SFTSV infection using an in vivo mouse model. Administration of antiserum protected against lethal infection with SFTSV. This treatment protected against clinical signs of disease due to non-lethal infection. Although ribavirin had no statistically significant effect, treatment still had a slight protective effect against SFTSV infection. MC treatment resulted in prolonged survival times during lethal infection. Although other agents had no significant protective effects, they did not show detrimental effects leading to disease progression. Our results suggest that antiserum treatment may be clinically useful for post-exposure prophylaxis against SFTSV infection.
4. Amuza Byaruhanga Lucky (Institute of Tropical Medicine, Nagasaki University)
"In vitro protracted cultivation of Plasmodium knowlesi for blood stage parasitology"
Introduction
In the past decade, concerted efforts and interventions have realised a significant decrease in global malaria incidence. At the same time, Plasmodium knowlesi has gained notoriety as one of the common causes of human malaria in Southeast Asia. The salient feature of this simian parasite is the ability to establish severe disease in both humans and monkeys through natural infection. In order to understand this unusual host sharing and adaptation to a wider niche of host cells, establishment of long term P.knowlesi in vitro culture that obviates the need for animal subjects and containment facilities is necessary. Firstly, we tried to adapt monkey-derived P. knowlesi H strain using published culture conditions for this parasite. However, we were able to maintain culture up to only 4 weeks. To overcome this challenge, multiple culture medium conditions were explored to optimise long term cultivation. To this end, we successfully established an in vitro culture condition that is able to support long term P. knowlesi H strain cultivation. By using this optimised condition, we also established culture-adapted P. knowlesi Hackeri strain for the first time. Further experiments to determine genetic factors mediating P. knowlesi host switch are ongoing.
 |
 |
| ● Date: | January 16 (Fri), 2015, 18:00 - 20:00 |
| ● Venue: | Lecture Hall |
1. Kochi Toyomane (Div. of Infection and Immunity, CZC, D1)
"Investigation of the molecular mechanism of AtxA-mediated virulence regulation on Bacillus anthracis"
Introduction
Anthrax is one of the fetal infectious diseases in human and animals, caused by Bacillus anthracis which is gram-positive spore forming bacterium. B. anthracis distributes worldwide including Japan, although no anthrax cases have been reported since 2001 in this country. The pathogenicity of B. anthracis depends on two major virulence factors, the anthrax toxin and the capsule. The anthrax toxin is composed of protective antigen, edema factor and lethal factor. These factors are coded on two plasmids, pXO1 for the toxin proteins and pXO2 for capsule synthases, and coordinately regulated at the transcription level by anthrax toxin activator gene, atxA. AtxA is a pXO1-encoded transcription factor and found in B. anthracis and the few B. cereus strains that harbor pXO1-like plasmid. It is known that AtxA activates not only expression of virulence gene but also that of the numerous genes on both chromosome and plasmids, in a CO2 dependent manner. However, activation mechanism of virulence gene expression by AtxA is not well understood. The objective of my research is to find molecular mechanism of AtxA-mediated virulence regulation, for which I am planning to analyze AtxA-DNA interations.
2. Suranji Wijekoon (Lab. of Veterinary Surgery, D1)
"Inhibitory Effects of Pentosan Polysulfate Sodium on Fos-Jun/AP1 Transcriptional Regulator Complex in Canine Bone Marrow Derived Osteoclasts In Vitro"
Introduction
Rheumatoid arthritis (RA) is generally a chronic, auto immune condition in which true cures or permanent remissions are unusual with the medication at the later stage of the disease. DMARDs that stand for disease-modifying antirheumatic drugs are an important part of an overall treatment plan. DMARDS exert minimal of anti- inflammatory effect and analgesic effect. Suppression of inflammation rather than immunosuppression in the early stages of the RA can result in substantial improvements in long-term outcomes. Recent years, it was newly recognized that PPS reduce pain and inflammation on osteoarthritis. But molecular biological mechanism of PPS as anti-bone resorptive agent on osteoclastogenesis and osteoclasts function in RA is not fully understood. The purpose of the current study is to investigate the mechanisms of action of PPS on inflammatory reaction of osteoclasts in vitro and compare with Heparin and PSGAG. PPS will be directly act on intracellular signaling pathway that play a pivotal role in many essential cellular processes such as proliferation and differentiation of osteoclasts which could be seen accelerating in the inflammation of RA process to produce osteoclasts. Bone marrow derived osteoclasts were treated during 14 days of osteoclastogenic process with PPS, Polysulphated glycosaminoglycan (PSGAG) and Heparin (0.2, 1 and 5 µg/ml) after treatment of human recombinant IL-1β (10ng/ml) as an inflammatory stimulator. Gene expression of osteoclasts differentiation markers (Carbonic anhydrase, calcitonine receptors, NFATc1, MMP9, c-Fos, RANK, Cathepsin K) and transcriptional regulator activator protein-1 was measured by using quantitative reverse transcription-polymerase chain reaction. Even there is no statistical significant on gene expression of calcitonine receptor by three treatments, highest concentration of PPS, PSGAG, Heparin significantly suppress the MMP9 gene and genomic expression of NFATC by PPS and Heparin (5µg/ml). Cathepsin K gene that is the major factor necessary for osteoclastic activity was significantly suppress by all concentration of PPS. Even Fos-Jun/AP1 transcriptional regulator complex was suppressed by heparin in dose dependent manner, PPS 1µg/ml, 5µg/ml concentrations was significantly affect AP1 activity. These data suggest that PPS capable to inhibit Fos-Jun/AP1 transcriptional regulator complex which is induced in RA by inflammatory cytokines and has a complex impact on osteoclast differentiation and production of soluble mediators of bone erosion.
3. Mizuho Suzuki (Lab. of Microbiology, D1)
"Exploring the mechanisms of interspecies transmission of H2 influenza viruses and vaccine preparation for a future pandemic"
Introduction
H2N2 influenza viruses caused a pandemic in 1957, however, the viruses have disappeared after 1968. Therefore infections of H2 viruses could be a pandemic in humans due to the lack of immunity against H2 viruses. H2 avian influenza viruses are circulating in birds and H2N3 virus was isolated from pigs. H2 viruses might transmit to humans from birds via pigs same as previous pandemic viruses. In order to prepare a future pandemic, vaccine preparation and exploring mechanisms of interspieces transmission are needed. In this study, genetic and antigenic analyses were conducted to prepare vaccines. And we conducted an experimental infection of pigs with H2 avian influenza viruses to reveal mechanisms of interspieces transmission.
The HA genes of H2 viruses were classified into two lineages; North American and Eurasian. The results of antigenic analysis suggested that antigenicity of North American and Eurasian viruses are closely related despite their genetic variation. As a future plan, viruses with high yield in embryonated eggs will be selected for vaccine strains. Three avian influenza viruses were inoculated to pigs and one virus was replicated in respiratory tract. Sequence analysis and measuring binding affinity to human-type receptor will be conducted.
4. Nipawit Karnbunchob (Div. of Bioinformatics, CZC, D2)
"Use of reciprocal best hits to explore the interspecies transmission of influenza A viruses"
Introduction
Influenza A virus is a zoonotic pathogen that infects avian and mammalian hosts. Genetic reassortment between human and avian viruses in pigs is responsible for the emergence of pandemic influenza viruses. Phylogenetic analysis is a conventional approach to study transmissions of the viruses. However, the growth in the amount of viral genetic information in the database increases the size of phylogenetic trees, creating technical difficulty in detecting the interspecies transmission. In this study, we employed the reciprocal best BLAST hits algorithm to detect interspecies transmission of avian viruses in pigs. Using nucleotide sequences of the hemagglutinin and neuraminidase of avian and swine influenza viruses, our method detected one hundred five possible avian-to-swine transmissions of H1N1, H1N2, H2N3, H3N2, H5N1, H5N2, H6N6, H7N2, H9N2, H10N5, H11N6, and H11N9. Thirteen results of them are consistent with the results from scientific papers that were published previously, suggesting that our method can correctly detect interspecies transmission.
5. Takeshi Kobayashi (Lab. of Pharmacology, D2)
"Identification of responsible mechanisms for the species difference of the effect of alpha2-agonists"
Introduction
Sedative effect of XYL has a clear species difference, very effective for cattle but less effective for pigs. DEX is effective for wide species of animals. So I hypothesized that α2A-AR subtype has some roles in this species difference.
Firstly I tried to compare the strictures of α2A-AR in human, mouse, cattle and pig. And then, I simulated the binding between α2A-AR and α2-agonists in each animals. As a result, the binding pocket of DEX was TM3, 5, 6 region in every animal's α2A-AR. Same results were obtained in XYL-α2A-AR expect the pig. The binding pocket of XYL was TM3, 4, 5 region in pig α2A-AR. This result suggests that the manner of binding would be responsible for the species difference of the effect of XYL.
So I'm planning to evaluate the changes of intracellar mechanisms evoked by XYL by using the expression systems. If the α2A-AR of each animals are expressed into AtT-20 cells, which endogenously express the Gi/o coupled GPCR, I will be able to evaluate the inhibition of Ca channel, activation of K channel and decrease of cAMP evoked by XYL. These mechanisms would be responsible for species difference.
 |
 |
| ● Date: | December 12 (Fri), 2014, 17:00 - 19:00 |
| ● Venue: | Lecture Hall |
1. Nao Eguchi (Div. of Global Epidemiology, CZC, D1)
"The mechanism of inhibition of influenza virus infection by M2e-specific antibody"
Introduction
The highly conserved nature of extracellular domain of M2 protein (M2e) among influenza A viruses makes it a potential candidate for the development of a universal vaccine. In a previous study it was found that an anti-M2e antibody (r2ss23), in the absence of neutralizing activity, inhibited virus release. Interestingly, the inhibitory mechanism still remains to be elucidated. Therefore, the present study is aimed at determining the inhibitory mechanism of r2ss23.
2. Shirin Akter (Lab. of Parasitology, D2)
"Epidemiological investigation of animal reservoir for visceral leishmaniasis (kala-azar) in Bangladesh"
Introduction
More than 60% of the world's visceral leishmaniasis (VL) cases are reported from Bangladesh, India and Nepal (WHO, 2005). The true incidence of VL in Bangladesh is close to 40,000-45,000 per year. Understanding of the epidemiological role of domestic and stray dogs as reservoir host for VL in Bangladesh is still very limited which is very important to improve the current control strategies. Serological and molecular tests were performed to investigate the dogs for the presence of Leishmania infection.
3. Chihiro Kanno (Lab. of Theriogenology, D1)
"Analysis of bovine sperm motility using computer-assisted sperm analysis (CASA) for the development of new criteria of sperm motility aimed at improvement of cryopreservation method"
Introduction
Sperm motility is very important for the fertilization. Computer-assisted sperm analysis (CASA) gives us many parameters related to motility for each sperm, but those detailed data have not used effectively. We do not know what the each parameter indicates and which kind of sperm related to fertilization. The aim of present study is to develop a profitable analyzing method of CASA data for the prediction of sperm fertility.
4. Dugar Delgermurun (Lab. of Pharmacology, D3)
"Mechanism of 5-HT outflow from glomus cells in the chick aorta induced by some chemical compounds"
Introduction
Antidepressants are believed to increase the extracellular level of serotonin (5-hydroxytryptamine; 5-HT) by inhibiting its reuptake into presynaptic cell. Amphetamine derivatives are capable of acting on amine transporters and then result in monoamine release from the neurons. Endogenous H2S is one of the key factors for rat carotid glomus cells responding to hypoxia. In this study, we investigated 5-HT outflow from 5-HT containing cells in the chick aorta induced by antidepressants, amphetamine derivatives and H2S.
5. Nesta Bortey-Sam (Lab. of Toxicology, D3)
"Concentrations of polycyclic aromatic hydrocarbon (PAH) metabolites in human and domestic animal urine, and their association with carcinogenicity"
Introduction
Polycyclic aromatic hydrocarbons (PAHs) are pollutants formed during incomplete combustion of organic materials. Many PAHs are mutagenic or carcinogenic. PAHs absorbed inside the body are metabolized (liver). There, they are converted into monohydroxylated PAHs and may be subsequently conjugated with glucuronide and sulfate before being eliminated in urine or bile. The aim of this study was therefore to determine concentrations of OH-PAHs, pyrene glucuronide and sulfate in cattle urine.
 |
 |
| ● Date: | November 7 (Fri), 2014, 18:00 - 20:00 |
| ● Venue: | Lecture Hall |
1. Yinghua Yang (Lab. of Theriogenology, D1)
"The effect of bone morphogenetic protein 4 (BMP4) on in vitro growth culture of oocytes derived from bovine early antral follicles"
Introduction
Theca cells are considered as an indispensable part to a follicle. BMP4, secreted exclusively by theca cells, is a regulator during folliculogenesis. However, the effect of BMP4 on in vitro growth (IVG) culture of oocytes from early antral follicles is still unclear. In present preliminary study, the effect of 2 doses of BMP4 added to IVG culture medium was investigated.
2. Eugene Chisela Bwalya (Lab.of Veterinary Surgery, D2)
"The effects of pentosan polysulphate or polysulphated glycosaminoglycan supplemented chondrogenic differentiation medium on chondrogenic differentiation of canine bone marrow-derived mesenchymal stem cells in three-dimensional alginate cultures"
Introduction
Articular cartilage is an avascular tissue that has a limited capacity for self-renewal and repair. Currently there is no effective treatment for osteoarthritis. Pluripotent mesenchymal stem cells offer the best alternative for cartilage tissue regeneration and repair of osteoarthritic defects by autologous implantation of differentiated stem cells. The use of PPS or PSGAG and 3-D alginate beads provides an intriguing novel approach to successful cartilage tissue engineering and warrants investigating.
3. Heidi Lynn Tessmer (Div. of Bioinformatics, CZC, D2)
"Tracking Influenza Epidemics using Bioinformatics Techniques"
Introduction
Influenza is a viral disease with annual epidemics resulting in 3 to 5 million cases and between 250,000 and 500,000 deaths [WHO]. Currently, the primary method of reducing influenza's impact is the administration of an annual vaccine. This research focuses on analyzing changes in the HA component of the H3N2 strain of human influenza to locate patterns in strain content and mutation with a goal of improving the accuracy of mutation prediction leading to a more effective influenza vaccine.
4. Jing Fang (Lab. of Veterinary Surgery, D3)
"Proliferation and differentiation of cultured canine chondrocytes in different concentrations of supplemental sera as nutrients"
Introduction
Serum is commonly used as a supplementation to cell culture media. In most cases of cell cultures, serum is added to culture media at concentrations of 5% or more. This may however cause to lose some phenotypic features of cells from cells physiologically maintained in low nutrients. The objective of this study is to investigate proliferation and differentiation capabilities in cultured articular chondrocytes under various densities of nutrients conditions.
5. Mohamed Elfeky (Lab. of Biochemistry, D3)
"Effect of adiponectin on LPS - induced HMGB1 release and HMGB1- induced inflammation"
Introduction
Adiponectin predominantly secreted from adipose tissue plays an important role in various physiological processes including lipid metabolism, insulin sensitization and anti-inflammatory responses. Although there are a great deal of interest that adiponectin attenuates inflammation, however, it is not known and there is no published report regarding whether adiponectin have a role in decreasing HMGB1 release and/or signaling.
 |
 |
| ● Date: | October 3 (Fri), 2014, 18:30 - 20:30 |
| ● Venue: | Lecture Hall |
1. Paulina Duhita Anindita (Div. of Molecular Pathobiology, CZC, D2)
"Development of Antiviral Therapy against Rabies Virus"
Introduction
Rabies is a fatal disease caused by rabies virus from genus Lyssavirus, family Rhabdoviridae. Rabies virus is usually transmitted to people by rabid animal bite, mostly from dogs but sometimes involving bats. Most cases ended with death. Currently, there is no effective drug against rabies. In this study, we attempt to establish a drug screening assay in order to find compounds that possess antiviral property against rabies and then evaluate drug candidates in vitro.
2. Yusuke Izumi (Lab.of Advanced Veterinary Medicine, D1)
"Isolation and characterization of tumor endothelial cells: investigation of immunosuppressive factor"
Introduction
Recently, a few reports indicated that the morphology and genetic expression of human and mouse tumor endothelial cells (TECs) might differ from normal endothelial cells (NECs). However, there is quite limited information about immunoreaction of TECs. In my preliminary experiment, I revealed that TECs expressed PD-L1 in immunohistopathological specimens. The purpose of my study is to isolate TECs and investigate immunosuppressive factors in TECs.
3. Jeewan Thapa (Div. of Bioresources, CZC, D2)
"Molecular characterization of Mycobacterium tuberculosis complex isolates from human and animal interface in Nepal"
Introduction
Tuberculosis is a chronic bacterial infection caused by members of M. tuberculosis complex (MTC). In Nepal, TB is prevalent in human, livestock and wild animals. Although, TB in humans has been studied, relatively comprehensively, studies of TB in livestock and wild animals are still in early stage and very few with molecular works. In this study, we are planning to study TB in human-animal interface with molecular epidemiological approach.
4. Zhifu Shan (Lab. of Veterinary Hygiene, D2)
"High throughput detection of PrPSc from prion-infected cells without PK-treatment: Cell-based ELISA for novel screening method for anti-prion compounds"
Introduction
Prion diseases are neurodegenerative disorders of humans and animals that are associated with conversion of normal prion protein (PrPC) to a disease-specific isoform of prion protein (PrPSc). No effective treatments are available for prion diseases to date. It is known that PrPSc consists of proteinase-sensitive (PrPSc-sen) and proteinase-resistant (PrPSc-res). Detection of PrPSc-sen is important because of the higher prion infectivity and conversion activity than PrPSc-res. However, most previous studies on screening anti-prion compounds only detected PrPSc-res after digesting PrPC and PrPSc-sen by proteinase K. Therefore, we established a novel, cell-based ELISA for screening of anti-prion compounds, in which PrPSc in the prion-infected cells were detected without proteinase K treatment.
5. Marvin A. Villanueva (Div. of Bioresources, CZC, D2)
"Understanding the circulation of Leptospira among water buffaloes in an intensive farm setting"
Introduction
Leptospirosis is considered the most widespread zoonosis worldwide. A wide range of animals harbor this bacteria and can be transmitted to human via infected urine. Our previous results revealed the presence of pathogenic leptospires in water buffalo, an indespensable livestock in the Philippines. In this study, we tried to understand how Leptospira circulate and maintained in an intensive farm setting, in order to control/prevent further spread of this disease which is of serious public health concern.
 |
 |
| ● Date: | September 5 (Fri), 2014, 18:00 - 20:00 |
| ● Venue: | Lecture Hall |
1. Jednipit Borthong (Div. of Bioinformatics, CZC, D1)
"Analysis of bacterial population in aquatic samples using 16s rRNA fragment through an ability of metagenomics strategy"
Introduction
Aquatic bacteria play significant roles for maintenance of elemental cycles and ecological systems. In order to comprehensively study whole population of bacteria in aquatic environments, a specific tool such as a metagenomic strategy can be used. This assay let us to elucidate the bacterial population using genetic material extracted from directly environmental samples and then sequenced through the sequencer machine. The sequences were compared the identity with the reference sequences in the database for identification of bacteria. Nowadays, metagenomic strategy becomes to the effective tool for studying the whole population of bacteria in aquatic environments.
2. Yuta Sakurai (Lab.of Biochemistry, D1)
"Age-related changes in the thermoregulatory strategy in harbor seal"
Introduction
Thick subcutaneous fat layer is important to maintain body temperature for seals. However, newborn seals store very little body fat and the thickness of their fat layer is extremely thin compared to that of adults. Thus, the other mechanisms may be involved in thermoregulation. In this study, I examined whether seals possessed brown adipose tissue (BAT) that is a site of cold-induced thermogenesis in small rodents.
3. Minato Hirano (Lab. of Public Health, D1)
"Study on genomic transportation-mechanism of neural infection of flaviviruses"
Introduction
The genus Flavivirus contains over 70 members, many of which are arthropod-borne pathogens distributed widely. Neurological diseases caused by encephalitic flaviviruses are severe and associated with high levels of mortality. However, detailed mechanisms of viral replication in the brain and features of viral pathogenesis remain poorly understood. To investigate the role of viral replication during flaviviral encephalitis, I focus on the molecular mechanism of viral genome-transportation in infected neuron.
4. Ayako Ujike (Lab. of Pharmacology, D2)
"The effect of hydrogen sulfide on enterochromaffin cells"
Introduction
Hydrogen sulfide (H2S) has been recently considered as a gasotransmitter. In the gastrointestinal tract, the amount of H2S is reported to increase in some diseases, such as colitis, suggesting that H2S acts as a pathological signal. Enterochromaffin (EC) cells, one of enteroendocrine cells, sense various luminal stimuli and regulate gastrointestinal function by 5-HT release. The purpose of this study is to reveal the relation between H2S signal and EC cells.
5. Marvin A. Villanueva (Div. of Bioresources, CZC, D2)
"Understanding the circulation of Leptospira among water buffaloes in an intensive farm setting"
Introduction
Leptospirosis is considered the most widespread zoonosis worldwide. A wide range of animals harbor this bacteria and can be transmitted to human via infected urine. Our previous results revealed the presence of pathogenic leptospires in water buffalo, an indespensable livestock in the Philippines. In this study, we tried to understand how Leptospira circulate and maintained in an intensive farm setting, in order to control/prevent further spread of this disease which is of serious public health concern.
 |
 |
| ● Date: | August 8 (Fri), 2014, 18:00 - 20:00 |
| ● Venue: | Lecture Hall |
1. Chihiro Ishii (Lab. of Toxiology, D1)
"Lead poisoning in raptors from Japan and source identification using lead stable isotope ratios"
Introduction
Lead poisoning is occurred among raptors because of consuming animals that were shot with Pb bullets or shot. In Hokkaido, the use of Pb bullets for deer hunting has been prohibited since 2001, however, one eagle died due to Pb poisoning in 2013. Few areas in other parts of Japan have regulations. In this study, we measured Pb concentrations and Pb stable isotope ratio to identify the source of poisoning.
2. Yogendra Shah (Div. of Bioresources, CZC, D1)
"Diversity of Mycobacterial interspersed repetitive units-variable number of tandem repeats among Mycobacterium tuberculosis Central Asian strain family isolates from Nepalese patients"
Introduction
Tuberculosis (TB) is the major public health problem in Nepal. Several South Asian countries has reported the predominant genotype of Mycobacterium tuberculosis (MTB) belonging to Central Asian Strain (CAS) family. Mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) is a reliable and reproducible method for differentiation of MTB isolates. However, information of its utility in determining the diversity of CAS strain is limited. In the present study, we evaluate and optimized the appropriate VNTR locus combination for the detailed genotyping of MTB strains of CAS family.
3. Naoya Maekawa (Lab. of Infectious Diseases, D1)
"Expression of PD-L1 on dog tumor cells and enhancement of IFN-γ production from tumor-infiltrating cells by PD-L1 blockade"
Introduction
Programmed death 1 (PD-1), an immunoinhibitory receptor, and programmed death ligand 1 (PD-L1), its ligand, together induce the "exhausted" status in antigen-specific lymphocytes and are thus involved in the immune evasion of tumor cells. In this study, dog PD-1 and PD-L1 were molecularly characterized, and their potential as therapeutic targets for dog tumors was discussed. The final goal of this study is to establish a novel immunotherapy against dog tumors.
4. Sawa Onouchi (Lab.of Anatomy, D2)
"Morphogenesis of the murine duodenojejunal flexure"
Introduction
The gastrointestinal tract has complex flexures for its compaction to fit into the abdominal cavity. These flexures are organized for the proper digestive system and determine gut unique morphology. Mammalian animals have species-common flexures in their gut morphology, and the duodenojejunal flexure is one of them. In this study, I focus on its morphogenesis as a model for flexure formation in intestinal development.
5. Nozomu Yokoyama (Lab. of Veterinary Internal Medicine, D2)
"Expression of claudins in the colorectal mucosa of inflammatory colorectal polyps in miniature duchshund"
Introduction
Inflammatory colorectal polyps (ICRPs) were recently recognized as a major cause of large bowel diarrhea in miniature dachshunds in Japan and are thought to be a novel form of inflammatory bowel disease (IBD). In human IBD, it has been suggested that derangements in tight junction (TJ) proteins may be involved in aberrant immune response in the intestine. The aim of this study is to determine the expression and distribution of TJ proteins claudins in the colorectal mucosa of ICRPs.
 |
 |
| ● Date: | July 11 (Fri), 2014, 18:00 - 19:40 |
| ● Venue: | Lecture Room 2 |
1. Yuji Wada (Div. of Molecular Pathobiology, CZC, D1)
"Study on Chikungunya virus; development of novel therapy and epidemiological survey"
Introduction
Chikungunya virus (CHIKV) is the cause of chikungunya fever, reemerging infectious disease, characterized by abrupt fever, rash, and acute and chronic joint pain. CHIKV is spreading all around the world including not only tropical areas that have often suffered from various insect-mediated infectious diseases but also temperate areas such as Italy and America. Because there are no approval drugs or vaccine, it is difficult to treat patient suffered from CHIKV and to prevent CHIKV spreading.
2. Yuri Sakaki (Lab. of Radiation Biology, D1)
"The effect of downregulation of DNA repair enzyme apurinic/apyrimidinic endonuclease 1 on cellular functions"
Introduction
Apurinic/apyrimidinic endonuclease 1 (APE1) plays an important role in the repair of DNA. Recent discoveries of new functions of APE1 show that APE1 is a multifunctional protein. However, it has not been well understood how these functions of APE1 affect cellular functions as a whole. In this study, the effect of APE1 downregulation on gene expression and cellular function was evaluated to reveal the biological role of APE1.
3. Minori Kuroda (Lab. of Veterinary Hygiene, D2)
"Involvement of astrocytes in neuropathogenesis of prion diseases"
Introduction
Activation of glial cells is a hallmark in prion diseases; however, the role of activated astrocytes remains to be elucidated. Thus my Ph.D. research focuses on understanding of astrocytes involvement in pathobiology of prion diseases. I am analyzing expression activation state of astrocytes from brains of prion-infected mice. Expression of CXCL10, which has multiple functions in neurodegeneration diseases was upregulated during 90 - 120 dpi.
4. Kiyeon Kim (Div. of Bioinformatics, CZC, D2)
"Inferring the population structure of Influenza A viruses with Tajima's D and its application to surveillance"
Introduction
Influenza has been big burden to public health all over the world. And thank to active surveillance and sequencing technique we can easily get huge amount of sequence data of Influenza A virus in public. Meanwhile, Tajima's D is a statistics that explain the state of population by using sequence dataset. Using public dataset, in this study, we tried to apply Tajima's D to comprehend epidemic situation in population.
 |
 |
| ● Date: | June 13 (Fri), 2014, 18:00 - 19:50 |
| ● Venue: | Lecture Hall |
1. Teppei Ikeda (Lab. of Anatomy, D1)
"Analysis of the functions of molecules that support bidirectional functions of caspases in skeletal muscle"
Introduction
Caspases are a family of cysteine proteases that are known to regulate apoptotic signal. Apoptosis by activation of caspases is strongly associated with embryonal development and regeneration in many organs. In previous study, we detected the activation of caspases in skeletal muscle of mouse embryo, whereas there were little apoptotic cells. In this study, we focus on apoptotic and non-apoptotic functions of caspases and clarify the mechanism of myogenesis and pathogenesis that caspases activation regulates in skeletal muscle.
2. Asami Nishimori (Lab. of Infectious Disease, D1)
"A simple and rapid method for diagnosis of bovine leukemia virus (BLV) infection"
Introduction
Bovine leukemia virus (BLV) is a retrovirus associated with persistent lymphocytosis and enzootic bovine leukosis. The recent research showed that the prevalence of BLV infections in Japanese dairy cattle was increased more than 8-fold in past thirty years. In this study, we attempted the establishment of a new method which was more simple and rapid for diagnosis of BLV infection, and compared its efficacy with that of a conventional method.
3. Keigo Fukano (Lab. of Biochemistry, D2)
"The proliferative ability of mature brown adipocytes and its regulation"
Introduction
There are two types of fat tissue in mammals, white adipose tissue and brown adipose tissue. It is generally assumed that the number of white adipocyte is controlled by the proliferation of pre-adipocytes and their differentiation, and mature adipocytes have no proliferative ability. However, little is known about the regulatory mechanisms of brown adipocyte number. To clarify the proliferative ability of mature adipocyte, we generated adipocyte-specific cell-cycle arrested mice.
4. Tomoyuki Yamaguchi (Div. of Bioresources, CZC, D2)
"The role of DNA gyrase and impact of amino acid substitution in DNA gyrase"
Introduction
DNA gyrase is a bacterial enzyme, which plays an essential role in bacterial replication and transcription by giving topological changes to the double strands of DNA. This enzyme is a target of quinolone, a family of commonly used antibacterial drugs. Amino acid substitution in the quinolone-binding site may confer quinolone resistance on bacteria. I will explain how DNA gyrase works and also talk about my research work on this enzyme.
 |
 |
| ● Date: | May 16 (Fri), 2014, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Balazs Oroszlany (Lab. of Toxicology, D3)
"Predictive Toxicology and modeling the reptile xenobioticmetabolism with transgenic yeast"
Introduction
Toxicology has to face two big challenges. One is to move away from animal testing (3R principles), and the other, at the same time, is to increase the throughput of pharmaceutical and chemical assays. This resulted in a fast development of human-focused *in silico* and *in vitro* screening methods. But how do these techniques translate into environmental risk assessment?
*Saccharomyces cerevisiae* based reptile xenobiotic metabolism models could answer that question.
2. Mohamed Moustafa (Lab. of Wildlife Biology and Medicine, D3)
"Epidemiology, isolation and identification of Anaplasma phagocytophilum from sika deer (Cervus nippon) in Hokkaido, Japan."
Introduction
Anaplasma phagocytophilumis a Gram-negative, obligate intracellular bacterium and causes human granulocytic anaplasmosis (HGA), which is a tick-borne rickettsial disease. In USA, the number of anaplasmosis cases reported to CDC has increased steadily since the disease became reportable, from 348 cases in 2000, to 1761 cases in 2010. Therefore, a study is required to understand the epidemiology of A. phagocytophilum and identify its strains in sika deer of Japan.
3. Siriporn Kongsoi (Div. of Bioresources, CZC, D3)
"Impact of mutations on DNA gyrase genes of Salmonella enterica"
Introduction
Quinolones are the first drug of choice for treatment of systemic salmonellosis that occurs in humans and animals. The prevalence of quinolone resistant Salmonella has become a concern for public health. Amino acid substitutions conferring resistance to quinolones in Salmonella enterica have generally been found within the quinolone resistance-determining regions (QRDRs) in the A subunit of DNA gyrase (GyrA).
 |
 |
| ● Date: | April 18 (Fri), 2014, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Wakako Furuyama (Div. of Global Epidemiology, CZC, D2)
"Study on host signaling pathway involved in antibody-dependent enhancenment of Ebola virus infection"
Introduction
Antibody-dependent enhancement (ADE) is a well-known phenomenon in which viral infectivity is increased by virus-specific antibodies in vitro. Ebolaviruses utilize viral glycoprotein-specific antibodies for their entry into cells through the interaction of the antibodies with cellular Fc receptors or complement component C1q and its receptors. Since these cellular molecules is known to activate various signaling pathways, we examined whether the ebolavirus ADE requires host cell signaling pathways through these molecules.
2. Tomokazu Tamura (Lab. of Micobiology, D3)
"Extra story of my PhD study－Immunopathogenesis of classical swine fever virus infection-"
Introduction
During evolution, viruses have acquired numerous mechanisms to evade or subvert key elements of the host viral response. For classical swine fever virus, the strategy to dampen the immune response by suppression of type I interferon and RNase activity was established. However, several questions are still remaining. I will show some pieces of data that could lead to understand virus-host interaction.
3. Katsuhisa Nagai (Lab. of Theriogenology, D3)
"The relationship between antral follicle counts (AFC) and quality of oocytes in cattle"
Introduction
Ovarian reserve is defined as the capability of ovaries to produce fertilizable oocytes, and antral follicle count (AFC) in ovaries is used as an indicator. Though reduced ovarian reserve is considered to impair the quality of oocytes in human, relationship between AFC and oocytes quality in cattle is still unclear. In this study, to clarify this relationship, fertilizabilities of oocytes derived from living cows with different AFC were examined.
 |
 |
| ● Date: | March 7 (Fri), 2014, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Motofumi Suzuki (Lab. of Radiation Biology, D1)
"Elucidation of the relationship between spindle assembly checkpoint and cellular radiosensitivity"
Introduction
In eukaryotes, the spindle assembly checkpoint (SAC) acts to maintain genome stability by delaying cell division until accurate chromosome segregation is ensured. Many studies have demonstrated that G1/S and G2/M checkpoints are two important checkpoints to determine the fate of irradiated cells. On the other hand, the role of SAC in cellular radioresponse remains elusive. The purpose of my study is to elucidate the relationship between SAC and cellular radioresponse.
2. Naganori Nao (Div. of Global Epidemiology, CZC, D2)
"Different pathogenicity of H5N1 highly pathogenic avian influenza viruses determined by a single amino acid in the M1 protein"
Introduction
Two highly pathogenic avian influenza virus strains, A/duck/Hokkaido/WZ83/2010 (H5N1) (WZ83) and A/duck/Hokkaido/WZ101/2010 (H5N1) (WZ101) showed different pathogenicity to chickens and ducks in spite of high nucleotide sequence identity of them. We revealed that the difference between WZ83 and WZ101 in the pathogenicity for chickens and ducks is determined only by the single amino acid in the M1 protein.
3. Keisuke Aoshima (Div. of Molecular Pathobiology, CZC, D3)
"Analysis of Reprogramming of Mouse Pronuclear Zygotes"
Introduction
Epigenetic reprogramming in zygotes is required to obtain the totipotency by erasing parental epigenetic marks such as DNA methylation and histone modifications. Although it is reported that reprogramming predominantly occurs in paternal pronucleus at the pronuclear stage soon after fertilization, the mechanism remains poorly understood. I would like to report on my progress so far in this meeting.
 |
 |
| ● Date: | February 7 (Fri), 2014, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Jemima Tiwaa Marfo (Lab. of Toxicology, D1)
"Establishment of diagnosis and elucidation of toxicological effects of subacute and chronic exposure to neonicotinoids"
Introduction
Neonicotinoid pesticides are systemic insecticides that possess nicotinic acetylcholine receptor agonist activity. Their importance in protecting crops and pest insect attack has been established. Nevertheless, there are some reports about toxicity of neonicotinoids to other animals and humans.
In regards to that, my aim is to investigate the toxicity of neonicotinoid pesticides to humans.
2. Nipawit Karnbunchob (Div. of Bioinformatics, CZC, D1)
"Role of α2A-adrenoceptor for the effects of dexmedetomidine and xylazine"
Introduction
Influenza A virus is a zoonotic pathogen that infects avian and mammalian hosts. Pigs are potential intermediate hosts for viral genetic reassortment between human and avian viruses. This genetic reassortment can led to new viral strains that might cause a pandemic. Thus, monitoring influenza A viruses in pigs play an important role to control the disease. The aim of this study is to clarify inter species transmission in avian, swine and human species.
3. Huy Duc Chu (Lab. of Microbiology, D1)
"Potency of an inactivated influenza vaccine prepared from A/duck/Mongolia/119/2008 (H7N9) against the challenge with A/Anhui/1/2013(H7N9) "
Introduction
H7N9 influenza virus infection in humans was recently reported in China on March 31, 2013. Characterization of the H7N9 influenza virus indicated that it has the potential to infect humans. The urgent need for the development of such a vaccine has been acknowledged by the WHO. A/duck/Mongolia/119/2008 (H7N9) was used to prepare an inactivated whole virus particle vaccine. The potency of this test vaccine was evaluated by a challenge with A/Anhui/1/2013 (H7N9).
 |
 |
| ● Date: | January 17 (Fri), 2014, 18:10 - 20:15 |
| ● Venue: | Lecture Hall |
1. Minori Kuroda (Lab. of Veterinary Hygiene, D1)
"Involvement of astrocytes in neuropathogenesis of prion diseases"
Introduction
Prion diseases are fatal neurodegenerative diseases that are characterized by PrPSc, neuronal vacuolation and microglial activation and astrocytosis. Recent studies in my lab suggested that after the appearance of PrPSc, astrocyte activation precedes microglial activation in the early stage of prion infection; however, the role of astrocytes in prion diseases remains to be elucidated. Thus I am focusing on elucidation of roles of astrocytes in neuropathogenesis of prion diseases.
2. Takeshi Kobayashi (Lab. of Pharmacology, D1)
"Role of α2A-adrenoceptor for the effects of dexmedetomidine and xylazine"
Introduction
Dexmedetomidine and xylazine, α2-adrenoceptor agonists, are used as the analgesics or sedatives for human and animals. Descending antinociception system and α2-adrenoceptor at spinal cord may involve these effects and α2A-adrenoceptor subtype has been reported to have a important role, but the mechanisms for these drugs are still unclear. So, I estimated the effects of dexmedetomidine and xylazine by in vitro and in vivo methods using C57BL/6J and α2A -adrenoceptor knockout mice.
3. Junpei Kimura (Lab. of Anatomy, D2)
"Elucidation of the pathogenesis of glomerular disease"
Introduction
Chronic kidney disease (CKD) is one of the most serious public health problems because it is strongly associated with not only end-stage renal disease (ESRD) but also cardiovascular diseases. Although membranous proliferative glomerulonephritis (MPGN) is a major primary cause of CKD, little has been known about the pathogenesis of MPGN. Here, I investigate the genetic factors contributing glomerular injuries using MPGN model mice.
 |
 |
| ● Date: | December 6 (Fri), 2013, 18:00 - 19:20 |
| ● Venue: | Lecture Hall |
1. Ryuji Kaede (Lab. of Biochemistry, D1)
"Roles of C1q/TNF related proteins in bovine mammary gland development and mastitis"
Introduction
Milk is important nutritious drink including proteins, lipids, calcium and so on, especially as safe drink in developing countries instead of contaminated water. Bovine mastitis damages mammary glands and significantly decreases milk production. C1q/tumor necrosis factor-related proteins (CTRPs), novel adipokines, paralogs of adiponectin, play an important role in anti-inflammation and so on. In terms of mammary gland, physiological roles regulated by CTRPs are completely unknown. So, we focus on roles of CTRPs in bovine mammary glands.
2. Hidehiro Yoshimura (Lab. of Infectious diseases, D1)
"Epidemiological research of Borrelia miyamotoi in Hokkaido"
Introduction
Tick parasites impact economically by reducing animal weight gain and livestock production. Ticks also transmit many kinds of pathogens that cause diseases which greatly impact human and animal health, or zoonotic diseases. Borrelia miyamotoi (B. miyamotoi) causes relapsing fever. B. miyamotoi was firstly isolated from Ixodes persulcatus in Hokkaido in 1995.In recent years, clinical cases in human were reported in Russia and North America, and field isolates in Russia were genetically close to those in Japan. No clinical cases in Japan have been reported, but B. miyamotoi may cause future outbreak in Japan. We researched the distribution of B. miyamotoi in Ixodes ticks in Hokkaido.
 |
 |
| ● Date: | November 8 (Fri), 2013, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Shan Zhifu (Lab. of Veterinary Hygiene, D1)
"Studies on the establishment of cell and immune therapy model for prion diseases"
Introduction
Bone marrow derived mesenchymal stem cells (MSCs) has been reported to show some therapeutic effects on prion infected mice. Anti-PrP antibodies or neurotrophic factors also have the neuroprotective effects on neurodegenerative diseases. Therefore, the present study is to build the MSC which can express anti-PrP antibody or neurotrophic factors and to find if they show the synergistic effects on therapy for prion diseases.
2. Fang Jing (Lab. of Veterinary Surgery, D2)
"Behavior of cultured canine chondrocytes under hyaluronic acid supplementation and low-density nutrients"
Introduction
Hyaluronic acid (HA) is a disease-modifying osteoarthritis drug to improve cartilage environment. In investigation of HA in vitro, 10% serum is commonly supplemented for maintenance of chondrocytes viability. However, chondrocytes in articular cartilage are in low-density nutrients. The aim of this study is to investigate effects of HA supplementation to culture media of chondrocytes on their proliferation, phenotype and differentiation under low-density nutrients.
3. Mohamed Sobhy (Lab. of Biochemistry, D2)
"Role of adiponectin in the regulation of obesity associated HMGB1 induced inflammation"
Introduction
Obesity is a metabolic disease of pandemic proportion. Obesity associated with a chronic inflammatory response characterized by increased expression of pro-inflammatory adipokines and decreased expression of the anti-inflammatory adipokine, adiponectin. The anti-inflammatory properties of adiponectin may be a major component of its beneficial effects on cardiovascular and metabolic disorders including atherosclerosis and insulin resistance. High-mobility group box 1 protein (HMGB1), a cytosolic ubiquitous protein stimulate necrosis-induced inflammation. Moreover, HMGB1 induces other cytokines. The identification of HMGB1 inhibitors is therefore of significant experimental and clinical interest.
 |
 |
| ● Date: | October 11 (Fri), 2013, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Sawa Onouchi (Lab. of Anatomy, D1)
"Novel Theory of Gut Morphogenesis -Spatiotemporal Gene Expression Designs Flexure Formation in Fetal Mice-"
Introduction
The gastrointestinal tract has complicated morphology, containing both species-common and species-specific features, meanings that gut morphogenesis is controlled by genetic programs. In this study, we are investigating murine gut morphogenesis especially that of the duodenojejunal flexure (DJF) based on spatiotemporal gene expression, different from the common theory depending on mesenteric tension. These findings will provide a crucial knowledge in the areas of developmental anatomy as well as regenerative medicine.
2. Tatsuyuki Osuga (Lab. of Internal Medicine, D2)
"Vitamin D status in canine heart disease"
Introduction
In humans with heart failure, vitamin D deficiency is common and associated with severe heart failure symptoms and short survival. In addition, recent evidence suggests that vitamin D supplementation in heart failure patients with vitamin D deficiency may prolong their survival. In dogs with heart disease, the status of vitamin D is unknown. The purpose of the study was to evaluate the status of vitamin D in canine heart disease.
3. Tomohiro Okagawa (Lab. of Infectious Diseases, D2)
"Bovine immunoinhibitory receptors: novel therapeutic targets for bovine leukemia virus infection in cattle"
Introduction
The immunoinhibitory receptors contribute to the immune evasion mechanisms of several viruses causing chronic infections. In bovine leukemia virus (BLV) infection, several inhibitory receptors played a critical role in functional exhaustion of T cell and disease progression. We are underway to establish the monoclonal chimeric antibodies specific to bovine inhibitory receptors and investigate these effects to develop a new therapeutic method for BLV infection.
 |
 |
| ● Date: | September 6 (Fri), 2013 18:00-19:20 |
| ● Venue: | Lecture Hall |
1. Delgermurun Dugar (Lab. of Pharmacology, D1)
"Mechanism of 5-HT release from glomus cells in the chick Aortic body (AB)"
Introduction
The glomus cells in chick aortic body (AB) sense changes in plasma PO2, PCO2 and pH and then release 5-hydroxytriptamine (5-HT). The purpose of this study is to reveal the effect of H2S and SSRIs drugs on 5-HT release from the glomus cells. In my preliminary experiments, we found that by both H2S and some SSRIs increased 5-HT release from the glomus cells in the chick AB.
2. Marvin A. Villanueva (Div. of Global Epidemiology, CZC, D1)
"Epidemiological study of leptospirosis among water buffalo (Bubalus bubalis)"
Introduction
Leptospirosis is a global re-emerging zoonotic infection caused by Leptospira spp. This study investigated the water buffalo, an indispensable animal in the Philippines that may contribute in maintaining and infecting leptospirosis to humans. Indirect ELISA system using recombinant outer membrane proteins were used in this study. Results showed all serum samples tested reacted to recombinant antigens used, suggesting that the disease is circulating and maintained within the herd, and may cause public health concern.
 |
 |
| ● Date: | August 9 (Fri), 2013, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Ayako Ujike (Lab. of Pharmacology, D1)
"The effect of hydrogen sulfide on enterochromaffin cells"
Introduction
Hydrogen sulfide (H2S), generally known as a toxic gas, is suggested to act as a gasotransmitter in mammalian body. High concentration of H2S is detected in large intestine and H2S production is reported to increase under pathological conditions such as colitis. The purpose of this study is to examine the effect of H2S on enterochromaffin cells which are distributed in the intestines and modulate intestinal function by 5-HT release.
2. Takahiro Hiono (Lab. of Microbiology, D2)
"Analyses of influenza virus receptors on the epithelial cells of chicken trachea"
Introduction
Wild ducks are the natural hosts of influenza A viruses. Although avian influenza viruses recognize SAα2,3Gal as virus receptors, chickens are rarely infected with viruses of duck origin. In order to better understand relationship between receptor specificity and host range of influenza virus in ducks and chickens, receptor binding specificity of the viruses and the distributions of receptors in tissues of ducks and chickens were investigated.
3. Saori Yano (Lab. of Physiology, D2)
"Characteristics of estrus cycle-dependent preference for male mice's odor"
Introduction
In mammals, individual information of a donor is included in odor. Female mice obtain information about males via the odor of the urine and choose mating partner. In a previous study, we found that the preference of females for odor of particular males is changed according to estrus cycle. The aim of this study is to investigate possible mechanism underlying regulation of the odor preference.
 |
 |
| ● Date: | July 5 (Fri), 2013, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Kim Kiyeon (Div. of Bioinformatics, CZC, D1)
"Inferring the population structure of viruses with Tajima's D and its application to global surveillance"
Introduction
Tajima's D is a statistic that discriminates mutation patterns in a set of nucleotide sequences. Based on the coalescent theory, Tajim's D tells us whether the population is expanding or declining and this property can be used for inferring the change of demography and evolution of population.
2. Bortey-Sam Nesta (Lab. of Toxicology, D1)
"Prevalence of Polycyclic aromatic hydrocarbons (PAHs) and their Metabolites in air, soil and urine of humans and domestic animals from Kumasi-Ghana"
Introduction
Polycyclic Aromatic Hydrocarbons (PAHs) are environmental pollutants found in air, soil, water and food. Most PAHs entering the environment are formed during burning of fossil fuel, wood etc. Many PAHs are mutagenic or carcinogenic. The aim of this study is to compare concentrations and abundances of PAHs and their metabolites (OH-PAHs), their contribution to the overall carcinogenic potential of PAH mixture and to assess the major sources of PAHs pollution in Kumasi.
3. Sarad Paudel (Lab. of Wild life Biology and Medicine, D2)
"Epidemiological and molecular studies of tuberculosis in captive elephants of Nepal"
Introduction
Epidemiological and molecular studies of tuberculosis (TB) were conducted in captive elephants of Nepal. Serological testing has shown that 25% elephants were infected with TB. Mycobacterium tuberculosis was isolated from three elephants having suspected TB lesions upon necropsy. Genotyping of isolates showed that the elephants were infected with local Mycobacterium tuberculosis strains prevalent among local population of Nepal. The results strongly suggest the transmission of TB from human to elephant.
 |
 |
| ● Date: | June 14 (Fri), 2013, 18:00 - 20:05 |
| ● Venue: | Lecture Hall |
1. Keigo Fukano (Laboratory of Biochemistry, D1)
"The mechanism of the disappearance of Brown Adipose Tissue (BAT)"
Introduction
BAT is one of two types of adipose tissue found in mammals. Its primary function is to generate body heat in animals or newborns that don't shiver. In human, it is known that the metabolic activity of BAT differs among individuals, being lower in older and obese individuals. But it is not known how it disappears. The aim of this study is to elucidate the mechanism of the disappearance of BAT.
2. Nozomu Yokoyama (Laboratory of Veterinary Internal Medicine, D1)
"The molecular mechanism and functional role of Toll-like Receptor in Canine Inflammatory Colorectal Polyps"
Introduction
Canine Inflammatory Colorectal Polyps (ICRP) in miniature dachshunds are recently recognized in Japan as a major cause of large bowel symptoms in this breed. In human, accumulating evidence indicate that Toll-like Receptors (TLRs) in intestinal epithelial cells play important roles in the pathogenesis of human IBD. The aim of my study is to examine the molecular mechanism and functional role of TLRs in the colonic epithelial cells of ICRP.
3. Junki Maruyama (CZC, Division of Global Epidemiology, D2)
"Characterization of the novel filovirus, Lloviu virus"
Introduction
Lloviu virus (LLOV) is a novel filovirus detected from bats in Europe. This virus is phylogenetically distinct from other filoviruses in the genus Ebolavirus or Marburgvirus, thus belongs to the newly proposed genus Lloviu cuevavirus in the family Filoviridae. Filovirus has a single envelope glycoprotein (GP) that plays major roles in the replication cycle and pathogenicity. In this study, we focused on GP to estimate the biological property of LLOV.
 |
 |
| ●日 時 | 平成25年5月17日（金）18：00－19：20 |
| ●場 所 | 講堂 |
【Title】
「Ovarian follicular reserve of cattle」
【Introduction】
A term "ovarian follicular reserve" is used to explain the capacity of the ovary providing oocytes with fertilizability resulting in a successful pregnancy. The number of small antral follicles in ovaries and anti-mullerian hormone produced by granulosa cells of growing follicles, both could be used as a marker of ovarian follicular reserve. So, we examined the utility of markers and the relationships between ovarian follicular reserve and fertility in cattle.
2ndYongjin Qiu（CZC, Division of Collaboration and Education, D3）
【Title】
「Tick microbial population analysis using 16S rRNA gene analysis with next generation sequencer」
【Introduction】
Ticks can transmit various pathogens. For example, nine spotted fever rickettsia species were identified as emerging agents recently. This fact indicates that ticks contain a large number of potential pathogens which have not been identified so far. 16S rRNA gene analysis is able to find unculturable microbes in environmental samples. The aim of this study is to analyze the tick microbes including potential pathogens by 16S rRNA gene analysis
 |
 |
| ●日 時 | 平成25年4月12日（金）18：00－20：10 |
| ●場 所 | 講堂 |
【Title】
「Implications of microparasites coexistence on the infection risk in Hokkaido wildlife」
【Introduction】
The study of tick borne pathogens (TBPs) in wildlife is extremely important because more than 70% of human emerging infectious diseases (EID) between 1940 and 2004 have their origin in wildlife. We believe that the coexistence of these pathogens at the same ecological system has created a network of interactions and co-infections between them inside the hosts. Our study will include the molecular diagnosis of different TBPs in Hokkaido wildlife.
2ndNaganori Nao （CZC, Division of Global Epidemiology, D2）
【Title】
「Mechanisms for the acquisition of high pathogenicity by avian influenza viruses」
【Introduction】
It is known that highly pathogenic avian influenza viruses (HPAIVs) evolve from low pathogenic precursor viruses maintained in wild aquatic birds, the natural host of influenza A viruses. While viruses of 16 hemagglutinin (HA; H1-H16) subtypes have been identified in the waterfowl reservoirs, the HA subtype of naturally isolated HPAIVs are restricted to only H5 or H7. The purpose of our research is to identify this restriction factors.
3rdJun Moriwaki （Laboratory of Wildlife Biology and Medicine, D2）
【Title】
「Development of studies on reproduction and behavior of brown bears with a goal of conservation of biodiversity」
【Introduction】
The Hokkaido brown bear (Ursus arctos yesoensis) is the largest terrestrial mammal in Hokkaido, Japan. Human-bear conflicts have become a social problem. For the conservation and management of wild brown bears, it is necessary to understand biological traits in brown bears, including population dynamics, etc. One of the biological factors which directly affect population dynamics is reproduction. For clarifying the reproductive rate of Hokkaido brown bears, we conducted two approaches.
 |
 |
| ●日 時 | 平成25年3月8日（金）18：00－20：10 |
| ●場 所 | 講堂 |
【Title】
「Survey of natural infection by Leishmania species in anthropophilic sand flies」
【Introduction】
Leishmaniasis is a vector-borne disease caused by the protozoan of genus Leishmania. The protozoan is transmitted by the female sand fly. Nearly 1,000 sand fly species has been described, but only a few are medically important. The present study employed the use of molecular biological method to evaluate natural Leishmania infection rates in man-biting sand flies.
2ndSangho Kim （Laboratory of Veterinary Surgery, D2）
【Title】
「Evaluate of association between the endoplasmic reticulum stress and canine osteoarthritis」
【Introduction】
Molecular mechanism of osteoarthritis (OA) is unclear, hence, therapeutic target of OA is not established. In this study we intend to confirm the hypothesis that excessive endoplasmic reticulum (ER) stress inducing cellular apoptosis plays a role in the pathology of cartilage degeneration. So, we investigate about evaluate of association between the ER stress and canine osteoarthritis.
3rdMakoto Kuroda （CZC, Division of Global epidemiology, D2）
【Title】
「Search for a receptor for filovirus」
【Introduction】
While several cellular molecules have been proposed to be filovirus receptors or co-receptors, the molecular mechanism of filovirus entry has not been fully understood. A monoclonal antibody M224/1, which was produced against some cellular molecule on the surface of Vero E6 cells, blocks infection with vesicular stomatitis virus pseudotyped with filovirus glycoprotein. In this study, we identified the target molecule of M224/1 by an expression cloning method using Vero E6 cDNA library.
 |
 |
| ●日 時 | 平成25年2月8日（金）18：00－19：20 |
| ●場 所 | 講堂 |
【Title】
「Analysis of Podocyte Injury in Murine Glomerulonephritis」
【Introduction】
Podocytes are highly differentiated cells lining the outside of glomerular capillaries. The podocyte foot processes form the slit diaphragm contributing to the regulation of glomerular filtration barrier. The effacement of foot processes are representative pathological features of podocyte injuries causing glomerular diseases with albuminuria. Although choronic glomerulonephritis (CGN) is a serious glomerular disease leading dialysis, little is known about the podocyte injuries. In the present study, we analyzed the pathological features of CGN model mice, especially focusing on the podocyte injuries.
2nd田村 友和 （微生物学教室, D1）
【Title】
「Identification of viral factors responsible for the pathogenicity of classical
swine fever virus in pigs」
【Introduction】
Numerous classical swine fever (CSF) virus strains show differences in CSF pathogenesis, resulting in clinical disease with different degrees of severity. The present study demonstrated that the amino acid residues in a structural protein and nonstructural proteins of CSF virus could function in a synergistic manner to determine the pathogenicity of the virus in pigs.
 |
 |
| ●日 時 | 平成25年1月11日（金）18：00－20：10 |
| ●場 所 | 講堂 |
【Title】
「Inhibition activity of quinolones against DNA gyrase of Campylobacter
jejuni」
【Introduction】
Quinolones are the antimicrobials most commonly used for treatment of adults with Campylobacter infections. Target of this drug in C. jejuni has been shown to be DNA gyrase. To investigate the interaction of C. jejuni gyrase with quinolones, we reconstituted in vitro the C. jejuni DNA gyrase by expressing the GyrA and GyrB subunits and evaluated the interaction of the enzyme with a large panel of quinolone with inhibition of DNA supercoiling assay..
2ndTatsuyuki Osuga （Lab. Of Internal medicine, D1）
【Title】
「Evaluation of left atrial phasic function by echocardiography in canine heart
disease」
【Introduction】
The left atrium modulates left ventricular filling through its 3 phasic function (reservoir, conduit, and booster pump function) to maintaining an optimal performance of the heart. In humans with heart diseases, the left atrial phasic function has been evaluated mainly by echocardiography and a strong correlation between left atrial dysfunction and the severity or the prognosis of the heart diseases has been described. In dogs with heart disease, the evaluation of the left atrial phasic function has not been reported and therefore the clinical usefulness of the assessment of the left atrial phasic function remains unknown. The purpose of the present study was to evaluate the clinical value of the assessment of the left atrial phasic function via echocardiography in canine heart disease.
3rdSayuri Nakamura （Lab. Of Comparative pathology, D2）
【Title】
「Pathologic features of astrocytic growth early after infection with fowl
glioma-inducing retroviruses」
【Introduction】
Fowl glioma is histopathologically characterized by multiple nodular astrocytic growths. This disease is caused by subgroup A of avian leukosis viruses, including fowl glioma-inducing virus (FGV). However, how and when astrocytes show neoplastic growth in this disease is not fully elucidated. Here, we investigated the relationship among viral replication, host immune responses and brain lesions in the initial infectious phase.
 |
 |
| ●日 時 | 平成24年12月7日（金）18：00－20：10 |
| ●場 所 | 第3講義室（新講義棟） |
【Title】
「Studies on the drug resistance in Salmonella Typhimurium, Salmonella
Enteritidis and Salmonella Choleraesuis」
【Introduction】
Salmonella gastroenteritis is generally a self-limiting illness; however, antimicrobial may be required in invasive or severe cases that may occur in some patients, particularly in vulnerable groups such as young children, elderly, and immune-compromised. Quinolones are the drug of first choice for treatment of systemic salmonellosis that occurs in humans and animals. However, resistance to these drugs has increased significantly in recent years.
2nd千原正尚 （解剖学教室, 博士課程2年）
【Title】
「Elucidation of the function of blood-testis barrier」
【Introduction】
In adult testes, differentiating meiotic and postmeiotic germ cells are separated from blood by the blood-testis barrier (BTB) formed by tight junctions between adjacent Sertoli cells. Although BTB has been considered to be essential for normal spermatogenesis, the function of BTB remains obscure. Here, we clarified the expression of BTB component proteins in adult mouse testis and investigated the spermatogenesis after BTB disruption.
3rd宮園耕介 （臨床分子生物学教室, 博士課程2年）
【Title】
「The elucidation of the alternative control mechanism of γ globin in bovine
globin switching」
【Introduction】
Globin switching in the birth period is a physiological response in which the globin chain substitution of the fetal γ-chain with the adult γ-chain. Bovine γ- and γ-globin possess the promoter sequences that are very similar to each other and the switching mechanism remains unknown. The purpose of the present study was to identify transcription factors and to analyze the mechanisms for differential promoter activation of γ- and γ-globin genes in cattle.
 |
 |
| ●日 時 | 平成24年11月2日（金）18：00－20：10 |
| ●場 所 | 第3講義室（新講義棟） |
【タイトル】
First steps in investigating the reptile xenobiotics metabolism: The metabolism of pyrene in freshwater turtles
【要旨】
Reptiles are one of the most endangered, and, at the same time, the most understudied group of vertebrates. To start investigating the xenobiotics metabolism in reptiles, as a first step, I have chosen freshwater turtles. Initial results indicates that turtles produce an unique metabolite distribution, which might be explained by the measured low activity of phase II enzymes.
第2演題 岡川朋弘（感染症学教室）
【タイトル】
ウシの免疫抑制受容体に対する阻害剤の開発および牛白血病ウイルス感染症の新規制御法への
応用
Investigation of immunoinhibitory receptors as novel therapeutic targets for bovine leukemia virus infection
【要旨】
牛白血病ウイルス (BLV) 感染症は、病態の進行に伴う免疫抑制を特徴とする慢性感染症である。我々はこれまでに、ウシの免疫抑制受容体を４種類同定し、これらの受容体がBLV感染症における免疫抑制に関与していることを報告した。本研究では、BLV感染症に対する新規治療法を開発することを目的に、これらの受容体を阻害するキメラ抗体を作製し、その治療効果を検討する。
Bovine leukemia virus (BLV) infection is characterised by immune supression in line with disease progression. Until now, we identified four immunoinhibitory receptors from cattle. Interestingly, we found that these inhibitory receptors play a critical role in immune supression during BLV infection. In this study, we establish the monoclonal chimeric antibodies specific to bovine inhibitory receptors, and investigate their effect to develop a new therapeutic method for BLV infection.
第3演題 足立真実（外科学教室）
【タイトル】
犬の血管肉腫転移に伴う血管内皮前駆細胞の変動
Functional changes in endothelial progenitor cells associated with metastasis of canine hemangiosarcoma
【要旨】
犬の血管肉腫は、高率に転移を引き起こす悪性腫瘍である。しかし、その転移機構は未だ解明されていない。我々は血管肉腫の転移は血管内皮前駆細胞によって引き起こされていると仮定し、血管肉腫罹患犬の末梢血中の血管内皮前駆細胞を検出することを目的として、これまでに得た成績を報告する。
Canine hemangiosarcoma (HSA) is a progressive malignant neoplasm because of a high metastasis. However, the underlying mechanism for this metastatic behavior of this tumor has not been elucidated yet. I hypothesized that this metastatic behavior was caused by EPCs. In this study, I am planning to detect circulating EPCs in the peripheral blood of dogs with HSA, and here we report some results.
 |
 |
| ●日 時 | 平成24年10月5日 (金) 18:00-20:05 |
| ●場 所 | 第3講義室（新講義棟） |
| ●演 題 | 「Characterization of antimicrobial resistance and class 1 integrons in |
| Escherichia coli isolated from shrimps and their environment」 | |
| Kanjana CHANGKAEW | |
| （人獣共通感染症リサーチセンター・国際疫学部門 D2） | |
| 「ニワトリ由来インフルエンザウイルスはカモ由来インフルエンザウイルスと | |
| 異なる構造のSAα2,3Galをレセプターとして認識している」 | |
| 日尾野 隆大（微生物学教室 D1） | |
| 「Tensin2変異マウスを用いた腎症抵抗性遺伝子の同定」 | |
| 佐々木 隼人（実験動物学 D2） |
 |
 |
| ●日 時 | 平成24年9月7日 (金) 18:00-20:10 | 引き続き ディスカッション20:10-20:40 |
| ●場 所 | 第2講義室（新講義棟） |
| ●演 題 | 「新規フィロウイルスの糖タンパク質GPの機能解析」 |
| 丸山 隼輝（人獣共通感染症リサーチセンター・国際疫学部門 D1） | |
| 「雌マウスの発情周期依存性の雄マウスに対する選好性のメカニズム解明 | |
| について」 | |
| 矢野 沙織（生理学教室 D1） | |
| 「In vitro production of embryo using sex-sorted spermatozoa」 | |
| 姜 成植（繁殖学教室 D2） |
 |
 |
| ●日 時 | 平成24年8月3日 (水) 18:00-19:30 | 小休憩の後、引き続き ディスカッション19:30-21:00 |
| ●場 所 | 講堂及び会議室（新講義棟） |
| ●演 題 | 「生体内における酵素を介した硫化水素(H2S)産生について」 |
| 宮本 亮（薬理学教室 D2） | |
| 「極東型ダニ媒介性脳炎ウイルスの強毒化に関わるウイルス側因子の特定」 | |
| 境 瑞紀（公衆衛生学教室 D2） |
