Application of complement component 4d immunohistochemistry to ABO-compatible and ABO-incompatible liver transplantation.

Antibody-mediated rejection (AMR) is difficult to diagnose after ABO-compatible or ABO-identical (ABO-C) liver transplantation. To determine whether complement component 4d (C4d) immunostaining would be useful for diagnosing AMR, we compared the results of C4d immunohistochemistry for allograft biopsy samples with assays for anti-donor antibodies performed at the time of biopsy. One hundred fourteen patients with ABO-C grafts and 29 patients with ABO-incompatible (ABO-I) grafts were included. Linear C4d endothelial staining (identifiable with a 4× objective lens) or staining seen in 50% or more of the portal tracts was considered positive. Five of the 114 patients (4%) with ABO-C grafts and 15 of the 29 patients (52%) with ABO-I grafts showed C4d positivity. In the ABO-C cases, C4d positivity in late biopsy samples (≥30 days after transplantation) was associated with stage 2 or higher fibrosis (METAVIR score; P = 0.01) and with the presence of donor-specific anti-human leukocyte antigen DR antibodies (HLA-DR DSAs) with a mean fluorescence intensity > 5000 according to the Luminex single-antigen bead assay (P = 0.04). Conversely, the presence of HLA-DR DSAs was associated with the presence of stage 2 or higher fibrosis, acute cellular rejection, and C4d positivity. During the 2-year follow-up, neither C4d positivity nor HLA-DR DSAs were related to graft loss. Among ABO-I patients, C4d positivity was not associated with allograft dysfunction or fibrosis. Only 3 of the 15 C4d-positive patients (20%) showed periportal hemorrhagic edema, which could be a histological sign of AMR in ABO-I grafts, and they were the only cases associated with elevations in anti-donor A/B antibody titers. In conclusion, C4d endothelial positivity among ABO-C patients is an uncommon event that could be associated with chronic graft damage with or without clinical AMR. C4d positivity is common among ABO-I patients and may not be associated with allograft dysfunction if alloantibody titers are not elevated

Retroperitoneal Laparoscopic Radical Nephrectomy for Renal Cell Carcinoma: A Report on 2 Initial Cases

We report our experience with retroperitoneal laparoscopic radical nephrectomy in 2 patients with renal cell carcinoma. In this procedure, a working space in the retroperitoneum is created using the blunt balloon dissection technique. Carbon dioxide insufflation is performed, and 4 trocars are inserted into the retroperitoneal cavity through the lateral abdominal wall. The kidney is removed together with the perirenal fat and Gerota's fascia in a muscle-splitting fashion. Using this procedure, a right nephrectomy was performed in a 65-year-old man with a 2.4-cm tumor and in a 54-year-old woman with a 3.5-cm tumor. Operative time was 220 min and 195 min, respectively, and estimated blood loss was 10 mL and 115 mL, respectively. There were no major perioperative complications. Although a long-term follow-up is necessary to evaluate the efficacy of this procedure, it will probably become a standard treatment modality for localized renal cell carcinoma

Pretransplant serum hepatitis C virus RNA levels predict response to antiviral treatment after living donor liver transplantation.

[Background]Given the limited efficacy and high adverse event rate associated with treatment of recurrent hepatitis C after liver transplantation, an individualized treatment strategy should be considered. The aim of this study was to identify predictors of response to antiviral therapy for hepatitis C after living donor liver transplantation (LDLT) and to study the associated adverse events. [Methods]A retrospective chart review was performed on 125 hepatitis C virus (HCV)-positive LDLT recipients who received interferon plus ribavirin and/or peginterferon plus ribavirin therapy at Kyoto University between January 2001 and June 2011. [Results]Serum HCV RNA reached undetectable levels within 48 weeks in 77 (62%) of 125 patients, and these patients were defined as showing virological response (VR). Of 117 patients, 50 (43%) achieved sustained VR (SVR). Predictive factors associated with both VR and SVR by univariate analysis included low pretransplant serum HCV RNA levels, a non-1 HCV genotype, and low pretreatment serum HCV RNA levels. In addition, LDLT from ABO-mismatched donors was significantly associated with VR, and white cell and neutrophil counts before interferon therapy were associated with SVR. Multivariate analysis showed that 2 variables–pretransplant serum HCV RNA level less than 500 kIU/mL and a non-1 HCV genotype–remained in models of both VR and SVR and that an ABO mismatch was associated with VR. No variables with a significant effect on treatment withdrawal were found. [Conclusions]Virological response to antiviral therapy in patients with hepatitis C recurring after LDLT can be predicted prior to transplant, based on pretransplant serum HCV-RNA levels and HCV genotype. LDLT from ABO-mismatched donors may contribute to more efficacious interferon therapy

Single Nucleotide Polymorphisms of the IZUMO Gene in Male Infertile Patients

IZUMOは精子頭部の膜表面に局在し、マウスの精子と卵の受精に必須なタンパク質である。日本人におけるIZUMO遺伝子の多型と精子形成の異常の関係を解析するために、我々は妊孕性の確認されたボランティアの男性１７２人と男性不妊症患者８９２人のゲノムDNAを用いてIZUMO遺伝子の多型と変異の検索を行った。ABI‐PRISM 3730xl Genetic Analyzer and SeqScape software （Applied Biosystems社）を用い解析を行った結果、妊孕性の確認されたボランティアの男性では検出されない一塩基置換を男性不妊症患者に見出した。初回の探索において、男性不妊症患者に４つの一塩基多型（４１７T>G（Cys１３９Trp），５８９A>T（Ser１９７Cys），９３９T>A（Phe３１３Leu），９４４G>A（Arg３１５Gln））が有意に検出された。これらの一塩基置換は、別のプライマーセットを用いた塩基配列解析では検出することができなかったが、一塩基多型と男性不妊症の関係の解析において重要な標的候補と考えられる。IZUMO is a surface protein on sperm and essential for egg fusion in mice. To investigate the possible association between variations in the IZUMO gene and impaired spermatogenesis in Japanese males, we screened for mutations in the human IZUMO gene using DNA from 892 sterile male patients and 172 proven-fertile male volunteers. Single Nucleotide Polymorphisms (SNPs) were identified in the heterozygous state in the infertile patients, and neither variant was identified in fertile subjects using an ABI-PRISM 3730xl Genetic Analyzer and SeqScape software (Applied Biosystems). Four single nucleotide polymorphisms (SNPs), 417 T>G (Cys 139 Trp), 589 A>T (Ser 197 Cys), 939 T>A (Phe 313 Leu), and 944 G>A (Arg 315 Gln), were found significantly more often in infertile subjects than in fertile controls in the first screening. Otherwise, the four SNPs were not detected by direct sequencing using the other primers. These results show that four SNPs exist as an analytical target of SNPs that associated with the male infertility n the IZUMO gene

[23-1]English Summaries of the Papers Contributed to Tonan Ajia Kenkyu (The Southeast Asian Studies) Vol.23, No.3 (Special Issue for Don Daeng : An Integrated Village Study in Northeast Thailand)

Tonan Ajia Kenkyu (The Southeast Asian Studies) = 東南アジア研究 (Japanese Journal of Southeast Asian Studies)[1]EDITOR'S NOTE … Hayao FUKUI[2]A Brief Look at the Village … Yoshihiro Kaida, Masuo Kuchiba[3]Typology of Rice Cultivation … Shuichi MIYAGAWA, Toshiro KURODA, Hiroyuki MATSUFUJI, Tomoo HATTORI[4]Instability of Rice culture … Yoshihiro KAIDA, Kazutoshi HOSHIKAWA, Yasuyuki KOHNO[5]POPULATION (I) … Hayao FUKUI[6]The Process of Emigration in Search of Good Land to Mo Nua Village, Udonthani Province … Yukio HAYASHI[7]An Economic Analysis of Endogenous Rural Economic Evolution and its Policy Implications … Hiroshi Tsujii[8]Kin Relationships and Kin Cooperation for Farming and Consumption … Masuo KUCHIBA, Takahiro TAKEMURA[9]Daily Activity Survey (1) … Satoshi KOIKE, Shinji SUWA, Haruo NOMA[10]Sharing of Merit and the Associated Social Relationships of Funeral Rites … Yukio HAYASHI[11]AN OVERVIEW ON NATURE, AGRICULTURE AND ECONOMY … Hayao FUKU

Collagen adhesion gene is associated with blood stream infections caused by methicillin-resistant Staphylococcus aureus

Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) causes hospital- and community-acquired infections. It is not clear whether genetic characteristics of the bacteria contribute to disease pathogenesis in MRSA infection. We hypothesized that whole genome analysis of MRSA strains could reveal the key gene loci and/or the gene mutations that affect clinical manifestations of MRSA infection. Methods: Whole genome sequences (WGS) of MRSA of 154 strains were analyzed with respect to clinical manifestations and data. Further, we evaluated the association between clinical manifestations in MRSA infection and genomic information. Results: WGS revealed gene mutations that correlated with clinical manifestations of MRSA infection. Moreover, 12 mutations were selected as important mutations by Random Forest analysis. Cluster analysis revealed strains associated with a high frequency of bloodstream infection (BSI). Twenty seven out of 34 strains in this cluster caused BSI. These strains were all positive for collagen adhesion gene (cna) and have mutations in the locus, those were selected by Random Forest analysis. Univariate and multivariate analysis revealed that these gene mutations were the predictor for the incidence of BSI. Interestingly, mutant CNA protein showed lower attachment ability to collagen, suggesting that the mutant protein might contribute to the dissemination of bacteria. Conclusions: These findings suggest that the bacterial genotype affects the clinical characteristics of MRSA infection. (c) 2019 The Author(s). Published by Elsevier Ltd on behalf of International Society for Infectious Diseases

A Single Nucleotide Polymorphism within the Novel Sex-Linked Testis-Specific Retrotransposed PGAM4 Gene Influences Human Male Fertility

The development of novel fertilization treatments, including in vitro fertilization and intracytoplasmic injection, has made pregnancy possible regardless of the level of activity of the spermatozoa; however, the etiology of male-factor infertility is poorly understood. Multiple studies, primarily through the use of transgenic animals, have contributed to a list of candidate genes that may affect male infertility in humans. We examined single nucleotide polymorphisms (SNPs) as a cause of male infertility in an analysis of spermatogenesis-specific genes.We carried out the prevalence of SNPs in the coding region of phosphoglycerate mutase 4 (PGAM4) on the X chromosome by the direct sequencing of PCR-amplified DNA from male patients. Using RT-PCR and western blot analyses, we identified that PGAM4 is a functional retrogene that is expressed predominantly in the testes and is associated with male infertility. PGAM4 is expressed in post-meiotic stages, including spermatids and spermatozoa in the testes, and the principal piece of the flagellum and acrosome in ejaculated spermatozoa. A case-control study revealed that 4.5% of infertile patients carry the G75C polymorphism, which causes an amino acid substitution in the encoded protein. Furthermore, an assay for enzymatic activity demonstrated that this polymorphism decreases the enzyme's activity both in vitro and in vivo.These results suggest that PGAM4, an X-linked retrogene, is a fundamental gene in human male reproduction and may escape meiotic sex chromosome inactivation. These findings provide fresh insight into elucidating the mechanisms of male infertility

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target