bioRxiv - Pathology最新文献

{"title":"Challenging the Conventional Treatment Initiation Paradigm: Early Detection of Irreversible Cellular Damage in Cardiac Biopsies of Fabry Disease Before the Formation of Gb3 Inclusion Bodies","authors":"Chung Lin Lee, Pei-Sin Chen, Yu-Ying Lu, Yu-Ting Chiang, Ching-Tzu Yen, Chun-Ying Huang, Yen-Fu Cheng, Hsiang-Yu Lin, Yun-Ru Chen, Dau-Ming Niu","doi":"10.1101/2024.06.28.601296","DOIUrl":"https://doi.org/10.1101/2024.06.28.601296","url":null,"abstract":"Background: Fabry disease (FD) is a lysosomal storage disorder impacting multiple organs, including the heart. We investigated whether early-stage globotriaosylceramide (Gb3) accumulation, before occurrence of inclusion bodies, could cause significant stress and irreversible damages of the cardiomyocytes in FD patients. To assess the cellular stress and irreversible damage of cardiomyocytes in FD during early-stage Gb3 accumulation before the occurrence of typical pathology.\u0000Methods: Immunofluorescent (IF) staining or Western blotting were performed on fibroblasts from FD patients and myocardial biopsies from G3Stg/GLAko mice and FD patients. Notably, all biopsies exhibited detectable Gb3 accumulation under IF but lacked typical FD (Gb3 inclusion body) pathology. Staining targeted nuclear factor-κB (NF-κB), interleukin-18 (IL-18), phospho-p42/44 mitogen-activated protein kinase (MAPK), and inducible nitric oxide synthase (iNOS) as inflammatory and oxidative stress markers. Alpha-smooth muscle actin (α-SMA) IF staining was conducted to detect myofibroblasts.\u0000Results: Fibroblasts from FD patients, in conjunction with cardiomyocytes from both G3Stg/GLAko mice and FD patients, exhibited significant accumulation of inflammatory markers such as NF-κB IL-18 and phospho-p42/44 MAPK, as well as the oxidative stress marker iNOS. Despite the absence of typical FD pathology, the presence of fibrosis was confirmed in myocardial biopsies from these patients through strong positive staining of α-SMA.\u0000Conclusions: Significant cellular stress and even irreversible damage may occur before the onset of typical pathological changes in cardiomyocytes of FD. Based on our findings, treatment should be initiated much earlier than we currently thought to prevent irreversible damage and improve the prognosis of FD patients.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"21 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507864","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Validation of morphological ear classification devised by principal component analysis using three-dimensional images for human identification","authors":"Hitoshi Biwasaka, Akihito Usui, Masataka Takamiya, Nikolaos Angelakopoulos, Roberto Cameriere, Akiko Kumagai","doi":"10.1101/2024.06.26.600836","DOIUrl":"https://doi.org/10.1101/2024.06.26.600836","url":null,"abstract":"This study attempts to classify ear morphology for human identification in forensic investigations by distinguishing between the upper auricle and lobule areas. A three-dimensional homologous model of the ear was created using 414 ear images of males aged 17–93 years reconstructed from computed tomography scans of forensic autopsy cases. Morphological changes were visualized using principal component analysis and areas of significant individual differences within the entire ear were identified. The classification criterion images for the upper auricle (ten images) and lobule (12 images) were developed by combining multiple principal component values: components 1–5 for the upper auricle and 1–6 for the lobule. Three-dimensional ear images of the upper auricle and lobule areas from 414 subjects were categorized using a measurement method based on the minimum distance between 5,507 corresponding points. The results indicate the applicability of the criterion images for the morphological classification of ears in this study.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"59 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141530868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Loss of asgr1a leads to the secretion of excess dietary cholesterol in zebrafish","authors":"Tabea O.C. Moll, Joshua T Derrick, Darby W Sweeney, Jeffrey Shin, Steven A Farber","doi":"10.1101/2024.06.26.600848","DOIUrl":"https://doi.org/10.1101/2024.06.26.600848","url":null,"abstract":"One of the major pathways to clear glycoproteins from circulation is via the liver-specific asialoglycoprotein receptor (ASGPR). Loss of asialoglycoprotein receptor 1 (ASGR1), the major subunit of ASGPR, was recently found to correlate with lower levels of plasma apolipoprotein B- containing lipoproteins (B-lps) and a profoundly reduced risk of cardiovascular disease in humans. We set out to identify the zebrafish ortholog of ASGR1 (asgr1a) and generated two independent mutations in asgr1a using CRISPR/Cas9. Neither asgr1a mutation displayed changes in larval, juvenile, and adult B-lp numbers or sizes. However, when challenged with a Western diet, asgr1a mutant zebrafish exhibit less hepatic steatosis and lower hepatic triglyceride levels compared to control animals. Instead, the excess dietary cholesterol was excreted. While these results do not explain the cardioprotective nature of ASGR1 in humans, they indicate the importance of ASGR1 in modulating whole animal cholesterol flux.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"21 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synovial fluid transcriptome dynamics in osteoarthritis progression: Implications in pathogenesis","authors":"Rinkle Sharma, Diksha Rana, Rahul Kumar, Sakshi Narula, Alpa Chaudhary, Bhavneet Kaur, Khushpreet Kaur, Mandeep Singh Dhillon, Devendra Chouhan, Uttam Chand Saini, Sadhna Sharma, Jyotdeep Kaur, Indu Verma","doi":"10.1101/2024.06.24.600143","DOIUrl":"https://doi.org/10.1101/2024.06.24.600143","url":null,"abstract":"Background: Osteoarthritis, a degenerative joint disease associated with various pathological manifestations in the joint including cartilage loss, alterations in subchondral bone and synovial inflammation. Objective: This study aimed to elucidate the transcriptional and molecular changes in synovial fluid associated with OA progression, focusing on differential gene expression and pathway enrichment across OA grades. Methodology: Patients with different OA grades were recruited from PGIMER, Chandigarh, following the KL classification. Microarray analysis was conducted to study the transcriptional profiles in different OA grades using a fold-change (FC) cutoff of 2 and a p-value cutoff of 0.05, followed by pathway analysis performed using GSEA and STRING database. Selected genes from microarray and pathway analysis were validated using qRT-PCR. Results: Microarray analysis reveals distinct gene expression patterns corresponding to different OA stages (KL grade 2 to KL grade 4). Notably, the upregulation of AMTN and DKK2, alongside the downregulation of MSLN, highlighted their roles in pathological mineralization and disrupted bone remodeling in OA. Pathway enrichment analysis revealed significant changes in immune response, inflammation related pathways and cellular processes such as autophagy and programmed cell death, indicating their involvement in disease progression. Furthermore, mitochondrial dysfunction and impaired autophagy were linked to increased inflammation in advanced OA. Conclusion: These findings suggest that targeting mineralization and inflammatory pathways could offer novel therapeutic avenues for OA management.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"190 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Negative Effects of Smoking Cigarettes on Morphological and Histological Damages to 18 and 21 Days Mouse Embryo Liver","authors":"zahra peyrovi, Soheil Azizi, Mohammad Hosseini","doi":"10.1101/2024.06.22.600226","DOIUrl":"https://doi.org/10.1101/2024.06.22.600226","url":null,"abstract":"Smoking cigarettes can lead to morphological and histological changes in several human and animal tissues. This paper describes the effects of CS exposure on the liver, weight, and length of mouse embryos. 42 NMRI adult mice were used for mating. Pregnant female mice were categorized into 3 groups: F (filtered), NF (non-filtered), and C (controlled). Each group included 14 female mice exposed to 12 cigarettes daily; 7 mice for 18 days and 7 mice for 21 days. In addition, only the group C were exposed to ambient air. Seven mouse embryos from each group were euthanized. Their livers were fixed for histological processing. The Livers of groups F & NF as compared with group C revealed changes in cellular architecture, centrilobular veins, inflammatory cells, kupffer cells, cytoplasmic vacuolation, hepatocytes necrosis, and decreased parenchymal cellularity. The average weight of embryo, liver, and CR in F & NF significantly reduced (p<0.05 and p<0.01).","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Surveillance and molecular characterization of banana viruses and their association with Musa germplasm in Malawi.","authors":"Johnny Isaac Gregorio Masangwa, Nuria Fontdevila Pareta, Philemon Moses, Eva Hřibová, Jaroslav Doležel, Isaac Fandika, Sebastien Massart","doi":"10.1101/2024.06.24.600487","DOIUrl":"https://doi.org/10.1101/2024.06.24.600487","url":null,"abstract":"Malawi has local banana germplasm preferred by its population. However, the epidemics of banana bunchy top disease, caused by the banana bunchy top virus (BBTV), is wiping out the most preferred germplasm and limiting its plantation. On the other hand, there needs to be more knowledge on viruses present in Malawi on banana crops. Therefore, a survey was conducted country-wide to characterize banana germplasm and evaluate the presence, incidence and prevalence of banana viruses. The survey covered four country zones, combined farmer's structured questionnaire and plant sampling. PCR products from infected germplasm were sequenced and aligned for each detected virus to build a phylogenetic tree. BBTV, BanMMV and BSV species were detected in Malawi. Malawi's BBTV isolates belonged to the Pacific Indian Ocean group only and BanMMV isolates clustered to three sub-branches. All the BSV species present in Malawi belonged to clade 1. Among the genetic groups of Musa, the identified banana germplasm belonged to AA, AAA, AB, AAB, and ABB groups with some germplasms unique compared to already genotyped germplasms. The ABB group was dominant in Malawi and was often infected by BSV species (probably originating from endogenous viral sequences), while BBTV more often infected the AAA group. Banana propagule sharing was the primary source of banana planting materials with a higher risk of spreading virus diseases. The survey underlined the need to set up a banana seed industry and policies that promote farmers' access to virus-tested planting materials, which will consequently prevent future virus epidemics.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"225 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Towards routine proteome profiling of FFPE tissue: Insights from a 1,200 case pan-cancer study","authors":"Johanna Tueshaus, Stephan Eckert, Marius Fraefel, Yuxiang Zhou, Pauline Pfeiffer, Christiane Halves, Federico Fusco, Johannes Weigl, Lisa Hönikl, Vicki Butenschön, Rumyana Todorova, Hilka Rauert-Wunderlich, Matthew The, Andreas Rosenwald, Volker Heinemann, Julian Walter Holch, Bernhard Meyer, Wilko Weichert, Carolin Mogler, Peer-Hendrik Kuhn, Bernhard Küster","doi":"10.1101/2024.06.21.600043","DOIUrl":"https://doi.org/10.1101/2024.06.21.600043","url":null,"abstract":"Proteome profiling of formalin-fixed paraffin-embedded (FFPE) specimens has gained traction for the analysis of cancer tissue for the discovery of molecular biomarkers. However, reports so far focused on single cancer entities, comprised relatively few cases and did not assess the long-term performance of experimental workflows. Here, we did so by analyzing 1,220 tumors from six cancer entities processed over the course of three years. Key findings include the need for a new normalization method ensuring equal and reproducible sample loading for LC-MS/MS analysis across cohorts, showing that tumors can, on average, be profiled to a depth of >4,000 proteins and discovering that current software fails to process such large data sets. We report the first comprehensive pan-cancer proteome expression resource for FFPE material comprising 11,000 proteins which is of immediate utility to the scientific community by way of a web resource. It enables a range of analysis including quantitative comparisons of proteins between patients or cohorts or the discovery of protein fingerprints representing the tissue of origin, or proteins enriched in certain cancer entities.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"50 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterizing neuroinvasion and neuropathology of SARS-CoV-2 by using AC70 human ACE2 transgenic mice","authors":"Jason Cheng-Chieh Hsu, Panatda Saenkham, Pinghan Huang, Cassio Pontes Octaviani, Aleksandra Drelich, Bi-Hung Peng, Chien-Te K Tseng","doi":"10.1101/2024.06.21.600068","DOIUrl":"https://doi.org/10.1101/2024.06.21.600068","url":null,"abstract":"COVID-19 presents with a plethora of neurological signs and symptoms despite being characterized as a respiratory disease, including seizures, anxiety, depression, amnesia, attention deficits, and alterations in consciousness. The olfactory nerve is widely accepted as the neuroinvasive route by which the etiological agent SARS-CoV-2 enters the brain, but the trigeminal nerve is an often-overlooked additional route. Based on this consensus, we initially conducted a pilot experiment investigating the olfactory nerve route of SARS-CoV-2 neuroinvasion via intranasal inoculation in AC70 human ACE2 transgenic mice. Notably, we found that the trigeminal ganglion is an early and highly efficient site of viral replication, which then rapidly spread widely throughout the brain where neurons were primarily targeted. Despite the extensive viral infection across the brain, obvious evidence of tissue pathology including inflammatory infiltration, glial activation, and apoptotic cell deaths were not consistently observed, albeit inflammatory cytokines were significantly induced. However, the expression levels of different genes related to neuronal function, including the neurotransmitter dopamine pathway as well as synaptic function, and markers of neuronal damage were altered as compared to mock-infected mice. Our findings suggest that the trigeminal nerve can be a neuroinvasive route complementary to the olfactory nerve and that the ensuing neuroinvasion presented a unique neuropathological profile. This study provides insights into potential neuropathogenic mechanisms utilized by coronaviruses.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"89 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141507861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Spatial Clustering Analysis with Spectral Imaging-based Single-Step Multiplex Immunofluorescence (SISS-mIF)","authors":"Tomohiko Nakamura, Noe Kaneko, Towako Taguchi, Kenji Ikeda, Moe Sakata, Miori Inoue, Tetsuro Kuwayama, Hirokazu Tatsuta, Iichiroh Onishi, Morito Kurata, Kazuhiro Nakagawa","doi":"10.1101/2024.06.17.597874","DOIUrl":"https://doi.org/10.1101/2024.06.17.597874","url":null,"abstract":"Precision medicine, anchored in spatial biology, is essential for the accurate diagnosis of cancer and prediction of drug responses. We have introduced the Spectral Imaging-based Single-Step Multiplex Immunofluorescence (SISS-mIF) technique, which leverages hyperspectral imaging to simultaneously capture fluorescence spectra. This approach automatically optimizes tissue autofluorescence spectra for each image, facilitating the use of fluorescent direct-labeled antibodies for multicolor staining in a single step. Unlike conventional methods, images are outputted as antibody counts rather than fluorescence intensity, allowing for consistent comparisons under different imaging conditions. We demonstrate that this technique allows for identical cell detection of CD3, CD5, and CD7 in T-cell lymphoma on a single slide. The utilization of fluorescent direct-labeled antibodies enables the triple staining of CD3, CD5, and CD7 without cross-reactivity, maintaining the same intensity as single stains. Moreover, we developed a joint Non-Negative Matrix Factorization-based Spatial Clustering Analysis (jNMF-SCA) with a modified spectral unmixing system, highlighting its potential as a supportive diagnostic tool for T-cell lymphoma.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly Repeatable Tissue Proteomics for Kidney Transplant Pathology: Technical and Biological Validation of Protein Analysis using LC-MS/MS","authors":"Rianne Hofstraat, Kristina Marx, Renata Blatnik, Nike Claessen, Aleksandra Chojnacka, Hessel Peters-Sengers, Sandrine Florquin, Jesper Kers, Garry Corthals","doi":"10.1101/2024.06.14.599091","DOIUrl":"https://doi.org/10.1101/2024.06.14.599091","url":null,"abstract":"Accurate pathological assessment of tissue samples is key for diagnosis and optimal treatment decisions. Traditional pathology techniques suffer from subjectivity resulting in inter-observer variability, and limitations in identifying subtle molecular changes. Omics approaches provide both molecular evidence and unbiased classification, which increases the quality and reliability of final tissue assessment. Here, we focus on mass spectrometry (MS)-based proteomics as a method to reveal biopsy tissue differences. For MS data to be useful, molecular information collected from formalin fixed paraffin embedding (FFPE) biopsy tissues needs to be consistent and quantitatively accurate and contain sufficient clinically relevant molecular information. Therefore, we developed an MS-based workflow and assessed the analytical repeatability on 36 kidney biopsies, ultimately analysing molecular differences and similarities of over 5000 proteins per biopsy. Additional 301 transplant biopsies were analysed to understand other physical parameters including effects of tissue size, standing time in autosampler, and the effect on clinical validation. MS data were acquired using Data-Independent Acquisition (DIA) which provides gigabytes of data per sample in the form of high proteome (and genome) representation, at exquisitely high quantitative accuracy. The FFPE-based method optimised here provides a coefficient of variation below 20%, analysing more than 5000 proteins per sample in parallel. We also observed that tissue thickness does affect the outcome of the data quality: 5 μm sections show more variation in the same sample than 10 μm sections. Notably, our data reveals an excellent agreement for the relative abundance of known protein biomarkers with kidney transplantation lesion scores used in clinical pathological diagnostics. The findings presented here demonstrate the ease, speed, and robustness of the MS-based method, where a wealth of molecular data from minute tissue sections can be used to assist and expand pathology, and possibly reduce the inter-observer variability.","PeriodicalId":501471,"journal":{"name":"bioRxiv - Pathology","volume":"37 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141515673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}