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Rxivist combines preprints from bioRxiv with data from Twitter to help you find the papers being discussed in your field. Currently indexing 83,434 bioRxiv papers from 359,709 authors.

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in category pathology

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301: Predictive factors in identifying operative risks in cholecystectomies.
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Posted to bioRxiv 05 Jul 2019

Predictive factors in identifying operative risks in cholecystectomies.
210 downloads pathology

Murat Kanlioz, Ugur Ekici

Purpose : This study aims to forecast findings showing the difficulty of operation in cholecystectomy through pre-operative examinations and reduce morbidity and mortality with the predictive data obtained. Materials and Methods: In the preoperative period, the followings were measured in patients who will undergo cholecystectomy: C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), WBC, Neutrophil ratio (NR), erythrocyte distribution range (RDW), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), total bilirubin (TB), direct bilirubin (DB). Following the preoperative ultrasound (USG), the patients were recorded in two groups as patients with "normal" and "increased" gallbladder wall thickness. Also, the patients were asked if they underwent ERCP and whether they received antibiotic treatment in the last 10 days due to their disease in the preoperative period. The appearance of the peroperative gallbladder was recorded in two groups as "has a normal appearance" or "edematous and/or adherent to peripheral tissues". Whether or not there is a correlation between the preoperative findings and peroperative appearance was evaluated. The recordings and analyzes were made using SPSS statistics program. Correlation between the data were analyzed by Chi-square test. p?0.05 was considered significant. Results: The study achieved statistically significant results for the correlation between the "gallbladder edema and/or adhesion to peripheral tissues" in the peroperative period and the following five parameters: increased WBC, increased NR, increased gallbladder wall thickness at USG, compulsory ERCP and receipt of antibiotic treatment for the disease in the last 10 days.(p?0,05). Conclusion: Taking into consideration the presence, in the preoperative period, of some or all of the five criteria -namely, increased WBC, increased NR, increased gallbladder wall thickness at USG, receiving antibiotic treatment for the disease in the last 10 days and undergoing ERCP- in patients with cholelithiasis for whom cholecystectomy is envisaged would make it easier to estimate the degree of difficulty of the surgery and the possibility of encountering difficult and complicated cases.

302: A Leptosphaeria maculans set of isolates characterised on all available differentials and used as control to identify virulence frequencies in a current French population
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Posted to bioRxiv 09 Jan 2020

A Leptosphaeria maculans set of isolates characterised on all available differentials and used as control to identify virulence frequencies in a current French population
210 downloads pathology

L. Bousset, M. Ermel, R. Delourme

The characterization of virulence frequencies has to be regularly updated to identify which genes are currently efficient and use this information to advise gene deployment by choosing varieties depending on the current composition of local pathogen population. In L. maculans on Brassica napus, because different genes were characterized by different teams, because new interactions are continuously identified and seed of differentials are difficult to obtain, we today still lack isolates characterized on all current resistance genes. On the one hand, we assembled a set of 12 isolates characterized on 13 of the 17 described resistance genes, having clearly compatible and clearly incompatible isolates for each interaction. This set can be used to characterize the L. maculans & B. napus interaction at cotyledon stage. Expanding the set of isolates with clearly virulent ones allowed us to detect inconsistent behaviour or intermediate (avirulent) phenotypes. On the other hand, we used this set of isolates as controls to identify virulence frequencies in a current French L. maculans population sampled in 2018 at Le Rheu. We provide the current status for 13 avirulence frequencies, including LepR1, LepR2 and LepR3 available in near isogenic lines of spring canola but not yet documented in France. Avirulence frequencies on the genes Rlm1, Rlm2, Rlm3, Rlm4, Rlm7, Rlm9 and LepR3 were low, indicating the lack of efficacy of these genes against the current population. In the opposite, all or most of isolates were avirulent for the genes Rlm5, Rlm6, Rlm10, Rlm11, LepR1 and LepR2. An optimistic point of view could conclude that there are ample resources for oilseed rape breeding. However, as compared to previous studies, so far all the resistance genes used on significant acreage without additional management practices have lost efficacy and only avirulences corresponding to resistance genes not deployed in France retain efficacy. While the call to wisely manage the available host resistance genes is not recent, it is still relevant. Adding, management practices to the deployment of resistance genes in order to reduce inoculum carry-over from one growing season to the next and to lower population sizes is key to maintain their efficacy over time.

303: DoUble resin Casting micro computed Tomography (DUCT) reveals biliary and vascular pathology in a mouse model of Alagille syndrome
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Posted to bioRxiv 21 Nov 2019

DoUble resin Casting micro computed Tomography (DUCT) reveals biliary and vascular pathology in a mouse model of Alagille syndrome
209 downloads pathology

Simona Hankeova, Jakub Salplachta, Tomas Zikmund, Michaela Kavkova, Noemi Van Hul, Adam Brinek, Veronika Smekalova, Jakub Laznovsky, Josef Jaros, Vítězslav Bryja, Urban Lendahl, Ewa Ellis, Edouard Hannezo, Jozef Kaiser, Emma R Andersson

BACKGROUND AND AIMS: Alagille syndrome, like several other liver diseases, is characterized by malformation of lumenized structures, such as the circulatory or biliary systems. Liver architecture has typically been studied through 2D sections and, more recently, using thick tissue sections combined with immunofluorescence. We aimed to develop a robust method to image, digitalize and quantify 3D architecture of the biliary and vascular systems in tandem. METHODS: The biliary and portal vein trees of the mouse liver were injected with Microfil resin, followed by microCT scanning. Tomographic data was segmented and analyzed using a MATLAB script we wrote to investigate length, volume, tortuosity, branching, and the relation between the vascular and biliary systems. Double resin casting micro computed tomography (DUCT) was applied to a mouse model for Alagille syndrome ( Jag1Ndr/Ndr mice), in which the biliary system is absent at postnatal stages, but regenerates by adulthood. Phenotypes discovered using DUCT were validated with cumbersome consecutive liver sections from mouse and human liver including patients with Alagille syndrome. RESULTS: DUCT revealed tortuous bile ducts either placed further from portal veins, or ectopically traversing the parenchyma and connecting two portal areas, in Jag1Ndr/Ndr mice. Furthermore, bile ducts either ended abruptly, or branched independently of portal vein branching, with bifurcations placed hilar or peripheral to portal vein branches. The branching defects, parenchymal bile ducts, and blunt endings were confirmed in patient samples. CONCLUSION: DUCT is a powerful technique, which provides computerized 3D reconstruction of casted networks. It exposes and quantifies previously unknown vascular and biliary phenotypes in mouse models, revealing new phenotypes in patients.

304: REVERSIBLE PULMONARY TRUNK BANDING. XI: MYOCARDIAL VASCULAR ENDOTHELIAL GROWTH FACTOR EXPRESSION IN YOUNG GOATS SUBMITTED TO VENTRICULAR RETRAINING
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Posted to bioRxiv 21 May 2019

REVERSIBLE PULMONARY TRUNK BANDING. XI: MYOCARDIAL VASCULAR ENDOTHELIAL GROWTH FACTOR EXPRESSION IN YOUNG GOATS SUBMITTED TO VENTRICULAR RETRAINING
208 downloads pathology

Renato S. Assad, Eduardo A. V. Rocha, Vera D. Aiello, Tiago A. Meniconi, Maria C. D. Abduch, Petronio G. Thomaz, Marcelo B. Jatene, Luiz F. P. Moreira

Background: Ventricle retraining has been extensively studied by our laboratory. Previous studies have demonstrated that intermittent overload causes a more efficient ventricular hypertrophy. The adaptive mechanisms involved in the ventricle retraining are not completely established. This study assessed vascular endothelial growth factor (VEGF) expression in the ventricles of goats submitted to systolic overload. Methods: Twenty-one young goats were divided into 3 groups (7 animals each): control, 96-hour continuous systolic overload, and intermittent systolic overload (four 12-hour periods of systolic overload paired with 12-hour resting period). During the 96-hour protocol, systolic overload was adjusted to achieve a right ventricular (RV) / aortic pressure ratio of 0.7. Hemodynamic evaluations were performed daily before and after systolic overload. Echocardiograms were obtained preoperatively and at protocol end to measure cardiac masses thickness. At study end, the animals were killed for morphologic evaluation and immunohistochemical assessment of VEGF expression. Results: RV-trained groups developed hypertrophy of RV and septal masses, confirmed by increased weight and thickness, as expected. In the study groups, there was a small but significantly increased water content of the RV and septum compared with those in the control group (p<0.002). VEGF expression in the RV myocardium was greater in the intermittent group (2.89% ± 0.41%) than in the continuous (1.80% ± 0.19%) and control (1.43% ± 0.18%) groups (p<0.023). Conclusions: Intermittent systolic overload promotes greater upregulation of VEGF expression in the subpulmonary ventricle, an adaptation that provides a mechanism for increased myocardial perfusion during the rapid myocardial hypertrophy of young goats.

305: Desmin is a modifier of dystrophic muscle features in Mdx mice
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Posted to bioRxiv 23 Aug 2019

Desmin is a modifier of dystrophic muscle features in Mdx mice
208 downloads pathology

Arnaud Ferry, Julien Messéant, Ara Parlakian, Mégane Lemaitre, Pauline Roy, Clément Delacroix, Alain Lilienbaum, Yeranuhi Hovhannisyan, Denis Furling, Arnaud Klein, Zhenlin Li, Onnik Agbulut

Duchenne muscular dystrophy (DMD) is a severe neuromuscular disease, caused by dystrophin deficiency. Desmin is like dystrophin associated to costameric structures bridging sarcomeres to extracellular matrix that are involved in force transmission and skeletal muscle integrity. In the present study, we wanted to gain further insight into the roles of desmin which expression is increased in the muscle from the mouse Mdx DMD model. We show that a deletion of the desmin gene (Des) in Mdx mice (DKO, Mdx:desmin-/-) induces a marked worsening of the weakness (reduced maximal force production) as compared to Mdx mice. Fragility (higher susceptibility to contraction-induced injury) was also aggravated and fatigue resistance was reduced in DKO mice. Moreover, in contrast to Mdx mice, the DKO mice did not undergo a muscle hypertrophy because of smaller and less numerous fibers, with reduced percentage of centronucleated fibres. Interestingly, Desmin cDNA transfer with adeno-associated virus in 1-month-old DKO mice and newborn Mdx mice improved muscle weakness. Overall, desmin plays important and beneficial roles on muscle performance, fragility and remodelling in dystrophic Mdx mice.

306: Detection of apoptosis and matrical degeneration within the intervertebral discs of rats due to passive cigarette smoking.
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Posted to bioRxiv 31 May 2019

Detection of apoptosis and matrical degeneration within the intervertebral discs of rats due to passive cigarette smoking.
208 downloads pathology

Masahiro Nakahashi, Mariko Esumi, Yasuaki Tokuhashi

Although low-back pain is considered to be associated with cigarette smoking, the influence of cigarette smoking on the intervertebral discs (IVD) has not been confirmed. We established a rat model of passive cigarette smoking-induced IVD degeneration, and investigated the cytohistological changes in the IVD and the accompanying changes in gene expression. IVD from rats exposed to 8 weeks of passive cigarette smoking were stained with Elastica van Gieson, and exhibited marked destruction of the supportive structure of the reticular matrix in the nucleus pulposus (NP). Positive signals on safranin O, alcian blue, type II collagen and aggrecan staining were decreased in the destroyed structure. Safranin O and type II collagen signals were also decreased in the cartilage end-plate (CEP) after 4- and 8-weeks of cigarette smoking. In the CEP, the potential for apoptosis was increased significantly, as demonstrated by staining for single-strand DNA. However, there were no signs of apoptosis in the NP or annulus fibrosus cells. Based on these findings, we concluded that passive cigarette smoking-induced stress stimuli first affect the CEP through blood flow due to the histological proximity, thereby stimulating chondrocyte apoptosis and reduction of the extracellular matrix (ECM). This leads to reduction of the ECM in the NP, destroying the NP matrix, which can then progress to IVD degeneration.

307: Towards Staining Independent Segmentation of Glomerulus from Histopathological Images of Kidney
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Posted to bioRxiv 29 Oct 2019

Towards Staining Independent Segmentation of Glomerulus from Histopathological Images of Kidney
208 downloads pathology

Robin Liu, Lu Wang, Jim He, Wenfang Chen

This paper introduces a detection-based framework to segment glomeruli from digital scanning image of light microscopic slide of renal biopsy specimens. The proposed method aims to better use the precise localization ability of Faster R-CNN and powerful segmentation ability of U-Net. We use a detector to localize the glomeruli from whole slide image to make the segmentation only focus on the most relevant area of the image. We explored the effectiveness of the network depth on its localization and segmentation ability in glomerular classification, and then propose to use the classification network with enhanced ability of localization and segmentation to construct and initialize a segmentation network. We also propose a weakly supervised training strategy to train the segmentation network by taking advantage of the unique morphology of the glomerulus. Both strong initialization and weakly supervised training are used to resolve the problem of insufficient and inaccurate data annotations and enhance the adaptability of the segmentation network. Experimental results demonstrate that the proposed framework is effective and robust.

308: Monodelphis domestica as a fetal intra-cerebral inoculation model for Zika virus pathogenesis
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Posted to bioRxiv 27 Sep 2019

Monodelphis domestica as a fetal intra-cerebral inoculation model for Zika virus pathogenesis
207 downloads pathology

John M. Thomas, Juan Garcia, Matthew Terry, Ileana Lozano, Susan M. Mahaney, Oscar Quintanilla, Dionn Carlo-Silva, Marisol Morales, John L. VandeBerg

Monodelphis domestica , also known as the laboratory opossum, is a marsupial native to South America.  At birth, these animals are developmentally equivalent to human embryos at approximately 5 weeks of gestation which, when coupled with other characteristics including the size of the animals, the development of a robust immune system during juvenile development, and the relative ease of experimental manipulation, have made M. domestica a valuable model in many areas of biomedical research.  However, their suitability as models for infectious diseases, especially diseases caused by viruses such as Zika virus (ZIKV), is currently unknown.  Here, we describe the replicative effects of ZIKV using a fetal intra-cerebral model of inoculation.  Using immunohistochemistry and in situ hybridization, we found that opossum embryos and fetuses are susceptible to infection by ZIKV administered intra-cerebrally, that the infection persists long term, and that the infection and viral replication consistently results in neural pathology and may occasionally result in global growth restriction.  These results demonstrate the utility of M. domestica as a new animal model for investigating ZIKV infection in vivo. This new model will facilitate further inquiry into viral pathogenesis, particularly for those viruses that are neurotropic, that may require a host with the ability to support sustained viral infection, and/or that may require intra-cerebral inoculations of large numbers of embryos or fetuses.

309: Measurement of Prostate Volume with MRI (A Guide for the Perplexed): Biproximate Method with Analysis of Precision and Accuracy
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Posted to bioRxiv 04 Dec 2019

Measurement of Prostate Volume with MRI (A Guide for the Perplexed): Biproximate Method with Analysis of Precision and Accuracy
205 downloads pathology

Neil F Wasserman, Eric Niendorf, Benjamin Spilseth

Purpose: To review the anatomic basis of prostate boundary selection on T2-weighted magnetic resonance imaging (MRI). To introduce an alternative 3D ellipsoid measuring technique that maximizes precision, report the intra- and inter-observer reliability, and to advocate its use for research involving multiple observers. Methods: A demonstration of prostate boundary anatomy using gross pathology and MRI examples provides background for selection of key boundary marks when measuring prostate volume. An alternative ellipsoid volume method is illustrated. An IRB approved retrospective study of 140 patients with elevated serum prostate specific antigen levels and/or abnormal digital rectal examinations was done with T2- weighted MRI applying the new (Biproximate) technique. Measurements were made by 2 examiners, correlated with each other for interobserver precision and with an expert observer for accuracy. Correlation statistics, linear regression analysis, and tests of means were applied using p≤0.05 as the threshold for significance. Results: Inter-observer correlation (precision) was 0.95 between observers. Correlation between these observers and the expert (accuracy) was 0.94 and 0.97 respectively. Intra-observer correlation for the expert was 0.98. Means for inter-rater reliability and accuracy were all the same (p=0.001). Conclusions: Anatomic foundations for the boundaries of the prostate are reviewed. Precision and accuracy of total prostate volume using an alternative method is reported and found to be reproducible.

310: Benzo(a)pyrene (B[a]P) and its adverse effects on male testies
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Posted to bioRxiv 27 Jun 2018

Benzo(a)pyrene (B[a]P) and its adverse effects on male testies
205 downloads pathology

Wei Liu, Aihua Gu

ABSTRACT It has been proved that Benzo(a)pyrene (B[a]P) is mutagenic in somatic cells, whereas the adverse effect of BaP on male reproduction remains unclear. To investigate whether it can pass through the blood-testis barrier (BTB) and its potential reproductive toxicology and molecular mechanisms, mice were exposed to B[a]P (there are two doses ,that is 13mg/kg body weight and 26 mg/kg body weight; three times per week) during 6 weeks and sacrificed 6 weeks after the final exposure to obtain B[a]P-exposed testis, blood and others. Electron microscopy analysis was performed to confirm whether the integrity of BTB and the ultra-structure changes in testes of B[a]P treated mice, which showed that the integrity of the BTB was disrupted, accompanied with the structure of sertoli cells seriously damaged, including the integrity of the nuclear membrane of the sertoli cells impaired and the basement membrane of the seminiferous tubules disrupted. X-ray imaging in vitro told us that BaP can overgo the BTB and gathered in the testis of mice. We found the significantly decreased expression of ZO-1, occludin, N-cadherin, vimentin and claudin-1 in the testes of B[a]P treated group by immunofluorescence detection. B[a]P induced BTB component protein decreased were also found in TM4 cells exposed to 5μmol/L B[a]P for 24h. We found a significantly decrease of testosterone level and a significantly increase of estrogen level in the serum of treated groups comparing with the control one by radioimmunoassay. TM4 cells, MLTC-1 cells and GC-2 cells was cultured with medium contains B[a]P. MTT Cell Proliferation and Cytotoxicity Assay ,cell apoptosis analysis, FACScan analyzer, We observed apparent increase of TM4 and GC-2 cells apoptosis after expose to B[a]P for 24h. B[a]P induced TM4 cell, GC-2 cell and MLTC-1 cell G2/M phase cell arrest. In conclusion, these results suggested that BaP has an adverse impact on male reproduction, it can cross the blood-testis barrier and damage it, the component proteins of the BTB significantly decreased, it can also produce adverse impact on male germ cells.

311: Cathepsin G Degrades Synovial Fluid Lubricin: Relevance For Osteoarthritis Pathogenesis
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Posted to bioRxiv 15 Oct 2019

Cathepsin G Degrades Synovial Fluid Lubricin: Relevance For Osteoarthritis Pathogenesis
204 downloads pathology

Shan Huang, Kristina A. Thomsson, Chunsheng Jin, Sally Alweddi, André Struglics, Ola Rolfson, Lena I Björkman, Sebastian Kalamajski, Tannin A. Schmidt, Gregory D. Jay, Roman Krawetz, Niclas G. Karlsson, Thomas Eisler

Lubricin (PRG4) is a mucin type protein that plays an important role in maintaining normal joint function by providing lubrication and chondroprotection. Improper lubricin modification and degradation has been observed in idiopathic osteoarthritis (OA), while the detailed mechanism still remains unknown. We hypothesized that the protease cathepsin G (CG) may participate in degrading lubricin in synovial fluid (SF). The presence of endogenous CG in SF was confirmed in 16 patients with knee OA. Recombinant human lubricin (rhPRG4) and native lubricin purified from the SF of patients were incubated with exogenous CG and lubricin degradation was monitored using western blot, staining by Coomassie or Periodic Acid-Schiff in gels, and with proteomics. Full length lubricin (~300 kDa), was efficiently digested with CG generating a 25-kDa protein fragment, originating from the densely glycosylated mucin domain (~250 kDa). The 25-kDa fragment was present in the SF from OA patients, and the amount was increased after incubation with CG. A CG digest of rhPRG4 revealed 135 peptides and 72 glycopeptides, and confirmed that the protease could cleave in different domains of lubricin. Our results suggest that synovial CG may take part in the degradation of lubricin, which could affect the lubrication of OA joints.

312: Degradations of tannin and saponin during co-composting of shell and seed cake of Camellia oleifera Abel
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Posted to bioRxiv 28 May 2019

Degradations of tannin and saponin during co-composting of shell and seed cake of Camellia oleifera Abel
204 downloads pathology

Jinping Zhang, Yue Ying, Xiaohua Yao

The degradation processes were studied in this paper of tannin and saponin during the co-composting of the shell and seed cake of Camellia oleifera Abel. Four treatments were designed with the dry weight of the seed cake accounting for 1/3(A1), 1/4(A2), 1/5(A3), and 1/10(A4) of the shell. The proportion of the seed cake is positively correlated with the duration of thermophilic phase, highest temperature and degradation rate of tannin and saponin whose maximum were in A1, but negatively correlated with C/N ratio and tannin content which were least in A1 of the final products. The content of saponin were all about 2% finally. The final content of saponin and tannin decreased 68.92-75.22% and 34.57-59.52%. The organic matters, total nutrient (N, P2O5 and K2O) increased with the rising proportion of the seed cake. Overall, the addition of the seed cake promoted the stability, fertilizer efficiency and safety of the co-compost product.

313: The P387 Thrombospondin-4 Variant Promotes Accumulation of Macrophages in Atherosclerotic Lesions
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Posted to bioRxiv 11 Jun 2019

The P387 Thrombospondin-4 Variant Promotes Accumulation of Macrophages in Atherosclerotic Lesions
203 downloads pathology

Santoshi Muppala, Mohammed Tanjimur Rahman, Irene Krukovets, Dmitriy Verbovetskiy, Elzbieta Pluskota, Aaron Fleischman, D. Geoffrey Vince, Edward F. Plow, Olga Stenina-Adognravi

Aims: Thrombopspondin-4 (TSP4) is a pro-angiogenic protein that has been implicated in tissue remodeling and local vascular inflammation. TSP4 and, in particular, its SNP variant, P387 TSP4, have been associated with cardiovascular disease. Macrophages are central to initiation and resolution of inflammation and development of atherosclerotic lesions, but the effects of the P387 TSP4 on macrophages remain essentially unknown. We examined the effects of the P387 TSP4 variant on macrophages in cell culture and in vivo in a murine model of atherosclerosis. Further, the levels and distributions of the twoTSP4 variants were assessed in human atherosclerotic arteries. Methods and Results: In ApoE-/-/P387-TSP4 knock-in mice, atherosclerotic lesions accumulated more macrophages than lesions bearing A387 TSP4. The levels of inflammatory markers were increased in lesions of ApoE-/-/P387-TSP4 knock-in mice compared to ApoE-/- mice. Lesions in human arteries from individuals carrying the P387 variant had higher levels of TSP4 and higher macrophage accumulation. P387 TSP4 was more active in supporting adhesion of cultured human and mouse macrophages in experiments using recombinant TSP4 variants and in cells derived from P387-TSP4 knock-in mice. Conclusions: TSP4 supports the adhesion of macrophages and their accumulation in atherosclerotic lesions. P387 TSP4 is more active in supporting these pro-inflammatory events in the vascular wall, which may contribute to the increased association of P387 TSP4 with cardiovascular disease.

314: Cyclophosphamide immunosuppressed Xid mice model clarify the protective role of B cells in experimental encephalitozoonosis
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Posted to bioRxiv 28 Sep 2018

Cyclophosphamide immunosuppressed Xid mice model clarify the protective role of B cells in experimental encephalitozoonosis
202 downloads pathology

Carla Renata Serantoni Moysés, Lidiana Flora Vidôto da Costa, Elizabeth Cristina Perez, José Guilherme Xavier, Diva Denelle Spadacci-Morena, Paulo Ricardo Dell’Armelina Rocha, Anuska Marcelino Alvares-Saraiva, Maria Anete Lallo

Encephalitozoon cuniculi is an intracellular pathogen that stablishes a balanced relationship with immunocompetent individuals, which is dependent of T lymphocytes activity. We previously showed X-linked immunodeficiency (XID – B cell deficient) mice are more susceptible to encephalitozoonosis and B-1 cells presence influences in the immune response. Because XID mice are deficient both in B-1 and B-2 cells, here we investigate the role of these cells against E. cuniculi infection using cyclophosphamide (Cy) immunosuppressed murine model to exacerbate the infection. XID mice presented lethargy and severe symptoms, associated with encephalitozoonosis and there was an increase in the peritoneal populations of CD8+ and CD4+ T lymphocytes and macrophages and also in the proinflammatory cytokines IFN-γ, TNF-α and IL-6. In BALB/c mice, no clinical signs were observed and there was an increase of T lymphocytes and macrophages in the spleen, showing an effective immune response. B-2 cells transfer to XID mice resulted in reduction of symptoms and lesion area with increase of B-2 and CD4+ T populations in the spleen. B-1 cells transfer increased the peritoneal populations of B-2 cells and macrophages and also reduced the symptoms. Therefore, the immunodeficiency of B cells associated to Cy immunosuppression condition leads to disseminated and severe encephalitozoonosis in XID mice with absence of splenic immune response and ineffective local immune response, evidencing the B-1 and B-2 cells role against microsporidiosis.

315: Clarification and confocal imaging of the non-human primate placental micro-anatomy
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Posted to bioRxiv 24 May 2018

Clarification and confocal imaging of the non-human primate placental micro-anatomy
202 downloads pathology

James A. Sargent, Victoria H.J. Roberts, Jessica Gaffney, Antonio E. Frias

Placental function is essential for the development of the fetus, and is - in part - related to the 3D arrangement of the villous and vascular geometry. Recent advances in tissue clarification techniques allow for deep high-resolution imaging with confocal microscopy without altering the spatial characteristics of the tissue. These image stacks can be analyzed quantitatively to provide insights regarding the villous and vascular micro-anatomy as well as the interrelationships between the two. However, such analyses require optimization of the tissue preparation, immuno-labeling, and clarification protocol in order to provide reliable results suitable for the detection of subtle differences in pathologic pregnancies. Placental and fetal development are similar between human and non-human primate pregnancies, with the latter serving as a reliable, validated, highly-controlled, well-characterized translational model for the former. We present a protocol for the preparation, immuno-labeling, and clarification of the non-human primate placenta optimized for confocal microscopy and subsequent quantification of the micro-anatomic structures.

316: A critical role of the T3SS effector EseJ in intracellular trafficking and replication of Edwardsiella piscicida in non-phagocytic cells
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Posted to bioRxiv 29 Nov 2018

A critical role of the T3SS effector EseJ in intracellular trafficking and replication of Edwardsiella piscicida in non-phagocytic cells
202 downloads pathology

Lingzhi Zhang, Jiatiao Jiang, Tianjian Hu, Jin Zhang, Xiaohong Liu, Dahai Yang, Yuanxing Zhang, Qin Liu

Edwardsiella piscicida (E. piscicida) is an intracellular pathogen within a broad spectrum of hosts. Essential to E. piscicida virulence is its ability to survive and replicate inside host cells, yet the underlying mechanisms and the nature of the replicative compartment remain unclear. Here, we characterized its intracellular lifestyle in non-phagocytic cells and showed that intracellular replication of E. piscicida in non-phagocytic cells is dependent on its type III secretion system. Following internalization, E. piscicida is contained in vacuoles that transiently mature into early endosomes, but subsequently bypasses the classical endosome pathway and fusion with lysosomes which depends on its T3SS. Following a rapid escape from the degradative pathway, E. piscicida was found to create a specialized replication-permissive niche characterized by endoplasmic reticulum (ER) markers. We also found that a T3SS effector EseJ is responsible for intracellular replication of E. piscicida by preventing endosome/lysosome fusion. Furthermore, in vivo experiments confirmed that EseJ is necessary for bacterial colonization of E. piscicida in both mice and zebrafish. Thus, this work elucidates the strategies used by E. piscicida to survive and proliferate within host non-phagocytic cells.

317: Developing a primary Paralichthys olivaceus gill epithelial cells as an in vitro model for propagation of VHSV show a corresponding increase in cell viability with increase in protein concentration in growth media
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Posted to bioRxiv 19 Jul 2019

Developing a primary Paralichthys olivaceus gill epithelial cells as an in vitro model for propagation of VHSV show a corresponding increase in cell viability with increase in protein concentration in growth media
198 downloads pathology

Sori Han, Jimin Hong, Jong-pyo Seo, Hetron Mweemba Munang’andu, In-kyu Yeo, Sung-hyun Kim

Background: Viral Hemorrhagic Septicemia Virus (VHSV) is a rhabdovirus that causes high mortalities linked to high economic losses in aquaculture. It has been grouped in four genotypes of which some do not easily propagate on continuous cell lines. As an alternative, the objectives of this study was to develop a primary gill epithelial cell (GEC) model from olive flounder (Paralichthys olivaceus) as an in vitro model for the propagation of VHSV. Results: Our findings show that the primary GECs developed herein are highly permissive to replication of the JF-09 genotype IVa strain leading to high cytopathic effect observed within 96 hours post virus inoculation. Our findings also show that the viability GECs produced herein corresponded with increase in the concentration fetal bovine serum in growth medium. We envision that GECs produced herein will heighten our understanding of immune mechanisms associated with virus entry on gill mucosal surfaces in flounder.

318: Hypertriglyceridemia and obesity exacerbate the course of SIRS induced by SAP in Rats
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Posted to bioRxiv 06 Jun 2019

Hypertriglyceridemia and obesity exacerbate the course of SIRS induced by SAP in Rats
198 downloads pathology

Kelei Hua, RuiXia Li, LiYing Cao, WanSheng Lao

The aim of the present study was to explore the mechanism underlying how HTG (hypertriglyceridaemia) and obesity exacerbate the course of the systemic inflammatory response syndrome (SIRS) induced by severe acute pancreatitis (SAP) in rats. Seventy-two rats were fed a normal or high-fat diet to induce HTG and obesity, and SAP was induced by retrograde injection of 5% sodium taurocholate solution at a volume of 1 ml/kg into the biliopancreatic duct. The injury to the pancreas was assessed by macroscopic observation, pancreatic histological evaluation and serum levels of amylase and lipase. SIRS was estimated by measuring SIRS scores and interleukin-6 (IL-6), tumour necrosis factor alpha (TNF-α) and interleukin-10 (IL-10) expression. The results showed that the SIRS scores and pancreatic histological scores increased significantly and the blood calcium level decreased significantly in the hypertriglyceridaemia SAP (HSAP) group compared with those of the SAP group. In addition, HTG and obesity significantly increased plasma levels of the proinflammatory cytokines IL-6 and TNF-α and significantly downregulated the proinflammatory cytokine IL-10. Our findings showed that HSAP rats exhibited more severe pancreatic injury and more serious SIRS scores than the SAP rats did. The underlying mechanism may be that HTG and obesity intensify early-stage SIRS by regulating the levels of inflammatory and anti-inflammatory cytokines.

319: New North American isolates of Venturia inaequalis can overcome apple scab resistance of Malus floribunda 821
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Posted to bioRxiv 07 Oct 2019

New North American isolates of Venturia inaequalis can overcome apple scab resistance of Malus floribunda 821
197 downloads pathology

David Papp, Jugpreet Singh, David Gadoury, Awais Khan

Apple scab, caused by Venturia inaequalis (Cke.) Wint., is a destructive fungal disease of major apple cultivars worldwide, most of which are moderately to highly susceptible. Thus, development of scab resistant cultivars is one of the highest priorities of apple breeding programs. The principal source of resistance for breeding programs has been the scab resistance gene Rvi6 that originated from the Japanese crabapple Malus floribunda (Sieb.) sel. 821. Isolates of V. inaequalis able to overcome Rvi6 have been identified in Europe, but have not yet been reported on the American continents. We recently discovered scab infection on M. floribunda 821 trees in a research orchard at Geneva, New York, USA, where approximately 10% of the leaves bore profusely sporulating apple scab lesions, many of which had coalesced to cover entire leaves. Chlorosis and pinpoint pitting symptoms typical of failed infections by V. inaequalis on hosts bearing the Rvi6 and Rvi7 genes were also observed. We assessed genetic diversity and population genetic structure of six V. inaequalis isolates collected from M. floribunda 821, one isolate from 'Nova Easygro', one isolate from 'Golden Delicious' and two isolates from Europe (11 isolates in total) using 16,321 genome-wide SNPs. Population genetic structure and PCA separated the isolates into distinct European and USA groups. The forgoing suggests that the new Rvi6 virulent isolates emerged within USA populations, rather than being transported from Europe. The overcoming of resistance in M. floribunda 821 but not in descendant cultivars suggests that durable resistance to apple scab will require a more comprehensive understanding of Rvi6 mediated resistance in diverse genetic backgrounds.

320: Loss of ELK1 has differential effects on age-dependent organ fibrosis and integrin expression
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Posted to bioRxiv 05 Sep 2019

Loss of ELK1 has differential effects on age-dependent organ fibrosis and integrin expression
197 downloads pathology

Jennifer T Cairns, Anthony Habgood, Rochelle C Edwards-Pritchard, Chloe Wilkinson, Iain D Stewart, Jack Leslie, Burns C Blaxall, Katalin Susztak, Siegfried Alberti, Alfred Nordheim, Fiona Oakley, R Gisli Jenkins, Amanda L Tatler

ETS domain-containing protein-1 (ELK1) is a transcriptional repressor important in regulating αvβ6 integrin expression. αvβ6 integrins activate the profibrotic cytokine Transforming Growth Factor β1 (TGFβ) and are increased in the alveolar epithelium in Idiopathic Pulmonary Fibrosis (IPF). IPF is a disease associated with ageing and therefore we hypothesised that aged animals lacking Elk1 globally would develop spontaneous fibrosis in organs where αvβ6-mediated TGFβ activation has been implicated. Here we identify that Elk1-knockout (Elk1-/0) mice aged to one year developed spontaneous fibrosis in the absence of injury in both the lung and the liver but not in the heart or kidneys. The lungs of Elk1-/0 aged mice demonstrated increased collagen deposition, in particular collagen 3α1, located in small fibrotic foci and thickened alveolar walls. Despite the liver having relatively low global levels of ELK1 expression, Elk1-/0 animals developed hepatosteatosis and fibrosis. The loss of Elk1 also had differential effects on Itgb1, Itgb5 and Itgb6 genes expression in the four organs potentially explaining the phenotypic differences in these organs. To understand the potential causes of reduced ELK1 in human disease we exposed human cells and murine lung slices to cigarette smoke extract which lead to reduced ELK1 expression which may explain the loss of ELK1 in human disease. These data support a fundamental role for ELK1 in protecting against the development of progressive fibrosis via transcriptional regulation of beta integrin subunit genes, and demonstrate that loss of ELK1 can be caused by cigarette smoke.

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