Diabetic condition induces hypertrophy and vacuolization in glomerular parietal epithelial cells

Diabetic nephropathy (DN) is accompanied by characteristic changes in the glomerulus, but little is known about the effect of diabetes on parietal epithelial cells (PECs). In this study, a descriptive analysis of PECs was undertaken in diabetic db/db mice and in diabetic patients. PEC hypertrophy was significantly more prominent in diabetic mice than in nondiabetic mice, and this was evident even at the early stage. Additionally, the number of vacuoles in PECs was markedly increased in diabetic mice, suggesting the presence of cellular injury in PECs in DN. Although rare, binuclear cells were observed in mice with early diabetes. In cultured PECs, a high glucose condition, compared with normal glucose condition, induced cellular hypertrophy and apoptosis. Flow cytometry showed that some PECs in the G0 phase reentered the cell cycle but got arrested in the S phase. Finally, in human diabetic subjects, hypertrophy and vacuolization were observed in the PECs. Our data showed that PECs undergo substantial changes in DN and may participate in rearrangement for differentiation into podocytes

Meeting Abstract Enhancing Automatic Classification of Hepatocellular Carcinoma Images through Image Masking, Tissue Changes, and Trabecular Features

Background Hepatocellular carcinoma (HCC) is a malignant tumor with hepatocellular differentiation and one of the most common cancers in the world. This type of cancer is often diagnosed when the survival time is measured in months causing high death rates Method We enhanced the classification process presented in [3] by including 11 features of tissue changes (i.e., features related to fatty change, cytoplasm colors, cell clearness index, and stroma) and 10 features of trabecular (e.g., nuclei-cytoplasmic ratio, irregularity of sinusoid, and trabecular arrangements). Furthermore, we apply a mask obtained by the stroma segmentation before calculating the 13 types of nuclear and structural features such that those features are derived from hepatocytes only, thus generating in total 177 features. The experiments were performed on a collection of region-ofinterest (ROI) images extracted from HE stained whole slide images (WSI), consisting of 1054 ROIs of HCC biopsy samples (504 negatives and 550 positives) and 1076 ROIs of HCC surgically resected samples (533 negatives and 543 positives). In the process, we made some combinations on the sets of features and sets of training data from both biopsy and surgery samples. As for the classification, we used 5-fold cross validation support vector machine (SVM) with LibSVM as our library. Results The results of classification experiment are summarized in Conclusion The combination of nuclear, trabecular, and other tissue features enables improved classification rate in HCC detection using SVM. Even though the image characteristics are different in biopsy and surgically resected samples, the same classification system gives good performance in both samples. The HCC classification scheme introduced in this paper is implemented in the prototype "feature measurement software for liver biopsy, " and the probability of HCC is visualized for every ROI in the WSI. It will support pathologists in the HCC diagnosis along with the quantitative measurements of tissue morphology

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

Maternal undernutrition aggravates renal tubular necrosis and interstitial fibrosis after unilateral ureteral obstruction in male rat offspring.

Maternal undernutrition is known to reduce glomerular number but it may also affect tubulointerstitium, capillary density, and response to oxidative stress. To investigate whether the latter elements are affected, we examined the response to unilateral ureteral obstruction (UUO), an established model of renal tubulointerstitial fibrosis, in the kidney of offspring from control and nutrient restricted rats. Six-week old male offspring from rats given food ad libitum (CON) and those subjected to 50% food restriction throughout pregnancy (NR) were subjected to UUO for 7 days. Body weight was significantly lower in NR. Systolic blood pressure and blood urea nitrogen increased similarly in CON and NR after UUO. Tubular necrosis in the obstructed kidney, on the other hand, was more extensive in NR. Also, the collagen area, a marker of fibrosis, of the obstructed kidney was significantly increased compared with the contralateral kidney only in NR. Capillary density was decreased similarly in the obstructed kidney of CON and NR compared with the contralateral kidney. Urine nitrate/nitrite, a marker of nitric oxide production, from the obstructed kidney was significantly increased in NR compared with CON. Nitrotyrosine, a marker of nitric oxide-mediated free radical injury, was increased in the obstructed kidney compared with the contralateral kidney in both CON and NR, but the extent was significantly greater in NR. In conclusion, more severe tubular necrosis and fibrosis after UUO was observed in NR, which was thought to be due to increased nitrosative stress

Color correction for automatic fibrosis quantification in liver biopsy specimens

Context: For a precise and objective quantification of liver fibrosis, quantitative evaluations through image analysis have been utilized. However, manual operations are required in most cases for extracting fiber areas because of color variation included in digital pathology images. Aims: The purpose of this research is to propose a color correction method for whole slide images (WSIs) of Elastica van Gieson (EVG) stained liver biopsy tissue specimens and to realize automated operation of image analysis for fibrosis quantification. Materials and Methods: Our experimental dataset consisted of 38 WSIs of liver biopsy specimens collected from 38 chronic viral hepatitis patients from multiple medical facilities, stained with EVG and scanned at ×20 using a Nano Zoomer 2.0 HT (Hamamatsu Photonics K.K., Hamamatsu, Japan). Color correction was performed by modifying the color distribution of a target WSI so as to fit to the reference, where the color distribution was modeled by a set of two triangle pyramids. Using color corrected WSIs; fibrosis quantification was performed based on tissue classification analysis. Statistical Analysis Used: Spearman′s rank correlation coefficients were calculated between liver stiffness measured by transient elastography and median area ratio of collagen fibers calculated based on tissue classification results. Results: Statistical analysis results showed a significant correlation r = 0.61-0.68 even when tissue classifiers were trained by using a subset of WSIs, while the correlation coefficients were reduced to r = 0.40-0.50 without color correction. Conclusions: Fibrosis quantification accompanied with the proposed color correction method could provide an objective evaluation tool for liver fibrosis, which complements semi-quantitative histologic evaluation systems

Meeting Abstract Development of a Prototype for Hepatocellular Carcinoma Classification Based on Morphological Features Automatically Measured in Whole Slide Images

Introduction The advent of new digital imaging technologies including high-throughput slide scanners is making a very compelling case as part of the clinical workflow. Tools developed for morphometric image analysis are accelerating the transition of pathology into a more quantitative science. The system for detection of regions suspected to be cancerous in gastric and colorectal tissue is already available. There is a real need for not only cancer detection but also quantification of histological features, because quantitative morphological characteristics can include important diagnostic and prognostic information. If an association between quantitative features and clinical findings is indicated, quantification of morphological features would be extremely useful to select the best treatment. Image measurement technology also has the potential for investigative pathology. We have developed a prototype system for both quantification of morphological features and automated identification of hepatocellular carcinoma (HCC) within whole slide images (WSI) of liver biopsy based on image recognition and measurement techniques. Our system displays quantified cell and tissue features as histogram, bar graph, and heat map on the screen. Displaying all features in such a unified visualization makes it easy to interpret quantitative feature. In this paper, we present a prototype designed specifically for morphological feature visualization in an easy-to-understand manner. Method Our system automatically analyzes WSI of liver biopsy stained by hematoxylin and eosin (H&E) stain, as shown in Results The placement of ROIs is in a fine balance, and quantitative morphological features are visually displayed, as shown i

Combined over-the-top reconstruction with posterolateral bundle remnant re-tensioning in pediatric anterior cruciate ligament reconstruction: A technical note

Epiphyseal injury, particular on femoral side, is a major concern in pediatric anterior cruciate ligament (ACL) reconstruction. Therefore, the over-the-top route (OTTR) method has frequently been selected in pediatric ACL reconstruction, with good clinical results reported. However, a cadaver study reported the inferior rotational stability of the OTTR method to that of anatomical single bundle reconstruction. In recent years, a new method of reconstruction, which involves the remnant being detached, re-tensioned, and re-attached, achieved good short-term results. We developed a surgical method to restore the remnant to the posterolateral (PL) bundle footprint and obtain rotational stability in patients, thereby preserving the remnant. We hypothesized that repairing the residual remnant to the PL bundle footprint in pediatric ACL reconstruction could achieve rotational stability. This report offers the surgical techniques for PL bundle tensioning repair using remnants in the pediatric ACL OTTR procedure