Epithelial Necrosis Research Articles

Seasonal development of the coccidian parasite Goussia bayae and hepatobiliary histopathology in white perch Morone americana from Chesapeake Bay.

The coccidium Goussia bayae infects the gallbladder and bile ducts of white perch Morone americana from Chesapeake Bay, USA. Seasonal changes in coccidian infections were analyzed from bile specimens of 1588 fish from the Choptank River during 2016-2018 using wet mount preparations with a Sedgwick-Rafter counting chamber. Histopathology of the gallbladder and liver was analyzed from a subset (n = 480) of these fish. Maximum parasite prevalence (100%) and intensities in the gallbladder occurred during the fish spawning season in March and April. Asynchronous coccidian development and prevalence of infections in fish increased gradually during autumn and winter, but coccidian intensity increased sharply 2-4 wk prior to the onset of fish spawning activity and decreased after spawning activity concluded. Sporulation was internal, and the gallbladder was the primary reservoir for oocysts. Two previously undescribed species of coccidia were observed in the intestine. Lesions in the gallbladder were rare and included cholecystitis and epithelial necrosis. Intrahepatic bile duct lesions were more common and included distension, cholangitis, epithelial erosion and necrosis, cholestasis, hyperplasia, and neoplasia. Cholangitis and necrosis of intrahepatic bile ducts were significantly associated with coccidial infections, while plasmodia of a myxosporean (spore morphology consistent with the genera Myxidium and Zschokella) were significantly associated with bile duct hyperplasia. Biliary neoplasia included cholangiomas (5% prevalence) and cholangiocarcinomas (1% prevalence). No association was detected between G. bayae and biliary neoplasms, but an association may exist between these lesions and the myxosporean plasmodia.

Skin burn and ocular damage by calcium oxide (virgin lime) in swines

ABSTRACT: An outbreak of skin burn and ocular damage by calcium oxide in swines is reported. The outbreak occurred in a rural property located in the municipality of Iuiu, Western Bahia, during the transport of 60 pigs from a recreation farm to a fattening farm. The observed macroscopic lesions were erythematous areas; formation of papules, vesicles, and in some cases, there was the formation of firm, parched, high brown plaques and in other cases detachment and loss of skin. His eyes were dull. Histological analysis of the skin revealed areas of necrosis of focally extensive clotting of the epidermis, and dermis, delimited in the paniculus, by cellular debris and rare neutrophils. Corneal necrosis with perforated ulcer formation, superficial epithelial necrosis, edema, and neutrophil infiltration of the cornea adjacent to the ulcer were revealed in the eye.

Pathological changes of the nasolacrimal duct in rabbit models of chronic dacryocystitis: correlation with lacrimal endoscopic findings.

The aim of this study was to explore the pathological changes of the nasolacrimal duct in rabbits with experimentally induced obstructive dacryocystitis in correlation with lacrimal endoscopic findings. The rabbit model of obstructive dacryocystitis was created by injecting 0.15ml of self-curing resin into the lacrimal duct. The control group received 0.15ml of normal saline. Within 16weeks after the obstructive, lacrimal endoscopy and pathological examination of the nasolacrimal duct were conducted at different time points of 1, 2, 4, 8, and 16weeks. In the control group, lacrimal endoscopy revealed pink and smooth mucosa; and the pathological analysis revealed an epithelial layer that was composed of superficial columnar cells and a deep basal epithelial layer. The experimental rabbits showed clinical manifestations of obstructive dacryocystitis a week after the injection of self-curing resin. At weeks 1 and 2, the lacrimal endoscopy showed mucosal hyperemia and hemorrhagic spots on the nasolacrimal duct; and the pathological features included epithelial cell swelling and inflammatory cell infiltration. At weeks 4 and 8, the experimental group showed alternatively red and white mucosa under the lacrimal endoscopy, and the pathological features included proliferative epithelium accompanied by papillary hyperplasia. At week 16, the experimental group showed pale and coarse mucosa and white membrane-like layer covering the mucosal surface, and the pathological features included epithelial necrosis, squamous metaplasia, and sub-epithelial fibrosis. The mucosa of the nasolacrimal duct showed different pathological features at different time points after lacrimal duct obstruction, which was well correlated with the endoscopic findings. It is possible to predict the pathological stages by the endoscopic observation in NLOD patients.

Pathological Changes of Copper Ion Poisoning in Mice

In this study, the clinical manifestations of copper poisoning and the pathological changes of tissues and organs were observed by copper sulfate. The experimental mice were randomly divided into four groups: control group (0 mg / kg), low dose group (50 mg / kg), medium dose group (100 mg / kg), high dose group (200 mg / kg), 6 rats in each group, the test period was 2 weeks. The results showed that the mortality rate in the high dose group was higher than that in the middle dose group and the low dose group. Pathological observation of high-dose group showed hepatic steatosis and granular degeneration, hepatocyte showed honeycomb appearance, low dose group and middle dose group hepatocyte enlargement, cytoplasm filled with red stained fine particles; low dose group In the renal tubular degeneration, renal tubular epithelial cell swelling, so that the lumen becomes smaller or narrow, interstitial telangiectasia, congestion. In the middle dose group, there were obvious degeneration of the renal tubules, and there were many red dye protein in the lumen. Part of the renal tubular wall cell structure was destroyed. High dose group of renal tubular epithelial necrosis, disintegration, fall off in the lumen. The results show that copper poisoning mainly damage the liver, kidney and other tissues and organs, eventually leading to morbidity and even death.

Preparation and characterization of pH-sensitive nanoparticles of budesonide for the treatment of ulcerative colitis.

ObjectiveThe aim of this study was to develop pH sensitive nanoparticles of budesonide for the treatment of ulcerative colitis.MethodsThe NPs system was characterized by the transmission electron microscopy (TEM), particle size, drug loading and encapsulation efficiency. In addition, in vitro drug release prop-erties and pharmacokinetics were also investigated in detail. The optimized formulation was examined for its in-vivo targeting potential using 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in a rat model.ResultsDynamic light-scattering results showed that the particle size of budesonide-Eudragit S100/poly(lactic-co-glycolic acid) nanoparticles was around 110.5 nm, with a polydispersity index of 0.098. Transmission electron microscopy images showed that BUD-ES100/PLGA NPs were spherical with uniform size and relatively smooth surfaces. In vitro release showed that BUD-ES100/PLGA NPs required minimal release of drugs during its transit in the stomach and the upper small intestine to ensure that a maximum dose reached the colon. After the pharma-codynamic treatment, the myeloperoxidase value of BUD-ES100/PLGA NPs was close to the normal group. The histopathological examination of rectum showed that no sign of damages such as epithelial necrosis and sloughing epithelial cells was detected.ConclusionOur findings suggested that BUD-ES100/PLGA NPs were a promising alternative to single pH-dependent systems for colitis therapy.

The impact of Clostridium butyricum MIYAIRI 588 on the murine gut microbiome and colonic tissue

BackgroundClostridium butyricum MIYAIRI 588 (CBM 588) is a probiotic bacterium that is used as an anti-diarrheal medicine in Japan. However, the impact of this probiotic on the gut microbiome has not been fully elucidated, especially, when used with antimicrobials. Material and methodsIn an in vivo study, CBM 588 monotherapy, clindamycin monotherapy, CBM 588 and clindamycin (combination therapy), or normal saline (control) was orally administered to mice for 4 days, and fecal samples were collected for 18 days to enumerate C. butyricum. We also extracted DNA from these fecal samples for metagenomics analysis by amplification of the V3-V4 region of the bacterial 16S rRNA gene and MiSeq Illumina sequencing. In addition, the concentrations of some short chain fatty acids were assessed in the fecal samples. A histological analysis was also conducted. ResultsOn day 4 (the last treatment day), there was no difference in the total counts of C. butyricum between the CBM 588 monotherapy and combination therapy groups (5.21 ± 0.78 vs. 5.13 ± 0.45 log10 cfu/g, p = 0.86). Clindamycin treatment resulted in dramatic increases in the phylum Firmicutes, especially Enterobacteriaceae, Clostridiaceae, Lactobacillus, and Enterococcus, compared with the other groups during the treatment period. CBM 588 treatment modified the bacterial community composition at lower phylogenetic levels. Some bacterial taxa, such as Bifidobacterium, Coprococcus, and Bacteroides, were significantly increased in the combination therapy group when compared with the other groups. In the metabolic analysis, CBM 588 enhanced lactic acid production. It also enhanced the efficiency of lactic acid use for the production of butyric acid. Only the clindamycin monotherapy group showed abnormal colon tissue, with superficial epithelial necrosis and the presence of inflammatory cells. ConclusionCBM 588 treatment modulated the gut microbiota composition under dysbiosis due to the use of an antimicrobial with strong activity against anaerobes and significantly reduced epithelial damage.

Effects of three commercial diets on growth, cataract development and histopathology of lumpfish (Cyclopterus lumpusL.)

Three duplicate groups of individually tagged lumpfish (mean initial weight: 20.0 ± 4.0 g) were fed three commercially available diets (A, B and C) for a period of 147 days. There were significant differences in growth rates between the groups with fish fed diet A having the highest growth rates whilst fish fed diet C achieved the lowest growth. Lumpfish fed diets A and B had lower prevalence and severity of cataracts compared with fish fed diet C, suggesting a possible dietary effect on cataract, alone or in combination with other environmental factors. There was a significant difference in the degree of liver vacuolization between the three dietary treatments at the end of the study period with fish fed diets A and B showing moderate to severe vacuolization compared with liver tissue from fish fed diet C, which had milder vacuolizations. There was little or no inflammation in pyloric caeca, midgut and hindgut and similarly for epithelial necrosis or vacuolization for all three dietary treatments at the end of the study. Fish fed diet A had the longest intestinal folds whilst fish fed diet C had the shortest.

Berberine Improves Mitochondrial Function in Colon Epithelial Cells to Protect L-Cells from Necrosis in Preservation of GLP-1 Secretion

Berberine (BBR) was reported to preserve the colon function in GLP-1 secretion in obese rodents through the butyrate-GPR43 pathway. However, the mechanism remains largely unknown except the pathway. Here, we tested BBR activity in the inhibition of clone epithelial necrosis in diet-induced obese mice. BBR was administrated in DIO mice through dietary supplementation for 8 weeks. Energy balance, insulin sensitivity and intestine mucosal function were monitored. Mitochondria were examined in the large intestine epithelial cells for structure and function to understand cell death. Mitochondrial structure was examined under the transmission electron microscopy and the function was determined using Seahorse XF24. Gut microflora was examined by Miseq sequencing and the fecal short chain fatty acids (SCFAs) were examined using gas chromatograph (GC). An increase in epithelial necrosis was observed in the colon mucosa together with a reduction in GLP-1 secretion and expression in DIO mice. Cristae loss and membrane rupture were observed in the mitochondrial structure of the colon epithelial cells. A disbalance of mitochondrial complexes was observed with an elevation in the complex I activity, and a fall in the complex II and IV activities. The abundance of short chain fatty acids were decreased along with the dysbiosis in the DIO mice. All of those alterations were corrected in DIO mice together with glucose disorder by the BBR treatment. The data suggest that BBR may protect the colon epithelial cells from necrosis in DIO mice to preserve GLP-1 secretion function. The effect was mediated by restoration of mitochondrial function from the disbalance among the complex I, II and IV. The direct and indirect effects of BBR are discussed. Disclosure J. Ye: None. J. Le: None. Y. Sun: None.

Caspase-3 Is a Pivotal Regulator of Microvascular Rarefaction and Renal Fibrosis after Ischemia-Reperfusion Injury.

Background Ischemia-reperfusion injury (IRI) is a major risk factor for chronic renal failure. Here, we characterize the different modes of programmed cell death in the tubular and microvascular compartments during the various stages of IRI-induced AKI, and their relative importance to renal fibrogenesis.Methods We performed unilateral renal artery clamping for 30 minutes and contralateral nephrectomy in wild-type mice (C57BL/6) or caspase-3-/- mice.Results Compared with their wild-type counterparts, caspase-3-/- mice in the early stage of AKI had high urine cystatin C levels, tubular injury scores, and serum creatinine levels. Electron microscopy revealed evidence of tubular epithelial cell necrosis in caspase-3-/- mice, and immunohistochemistry showed upregulation of the necroptosis marker receptor-interacting serine/threonine-protein kinase 3 (RIPK3) in renal cortical sections. Western blot analysis further demonstrated enhanced levels of phosphorylated RIPK3 in the kidneys of caspase-3-/- mice. In contrast, caspase-3-/- mice had less microvascular congestion and activation in the early and extension phases of AKI. In the long term (3 weeks after IRI), caspase-3-/- mice had reduced microvascular rarefaction and renal fibrosis, as well as decreased expression of α-smooth muscle actin and reduced collagen deposition within peritubular capillaries. Moreover, caspase-3-/- mice exhibited signs of reduced tubular ischemia, including lower tubular expression of hypoxia-inducible factor-1α and improved tubular injury scores.Conclusions These results establish the pivotal importance of caspase-3 in regulating microvascular endothelial cell apoptosis and renal fibrosis after IRI. These findings also demonstrate the predominant role of microvascular over tubular injury as a driver of progressive renal damage and fibrosis after IRI.

The Ex vivo Eye Irritation Test (EVEIT) model as a mean of improving venom ophthalmia understanding

Snakes belonging to the genus Naja (Elapid family), also known as “spitting cobras”, can spit venom towards the eyes of the predator as a defensive strategy, causing painful and potentially blinding ocular envenoming. Venom ophthalmia is characterized by pain, hyperemia, blepharitis, blepharospasm and corneal erosions. Elapid venom ophthalmia is not well documented and no specific treatment exists. Furthermore, accidental ejection of venom by non-spitting vipers, as Bothrops, also occurs. The Ex vivo Eye Irritation Test model (EVEIT) has enabled important progress in the knowledge of chemical ocular burns. Considering the lack of experimental animal model, we adapted the EVEIT to study venom ophthalmia mechanisms. Ex vivo rabbit corneas were exposed to venoms from spitting (Naja mossambica, Naja nigricollis) and non-spitting (Naja naja, Bothrops jararaca and Bothrops lanceolatus) snakes, and rinsed or not with water. The corneal thickness and the depth of damage were assessed using high-resolution optical coherence tomography (HR-OCT) imaging and histological analysis. All Naja venoms induced significant corneal edema, collagen structure disorganization and epithelial necrosis. Corneas envenomed by African N. mossambica and N. nigricollis venoms were completely opaque. Opacification was not observed in corneas treated with venoms from non-spitting snakes, such as the Asian cobra, N. naja, and the vipers, B. jararaca and B. lanceolatus. Moreover, Bothrops venoms were able to damage the epithelium and cause collagen structure disorganization, but not edema. Immediate water rinsing improved corneal status, though damage and edema could still be observed. In conclusion, the present study shows that the EVEIT model was successfully adapted to set a new experimental ex vivo animal model of ophthalmia, caused by snake venoms, which will enable to explore new therapies for venom ophthalmia.

Photosensitization in Cattle Caused by Spontaneous and Experimentally Ingestion of Stryphnodendron fissuratum

Background: Stryphnodendron fissuratum is a tree from the Brazilian Cerrado. Its fruit is toxic to cattle and can cause clinical digestive signs, hepatogenous photosensitization, and abortion. Cases of poisoning in cattle, goats and guinea pigs have been experimentally reproduced; however, photosensitization could not be reproduced. The aim of this work was to describe an outbreak of natural poisoning and experimental reproduction in cattle, both with hepatogenous photosensitization.Materials, Methods & Results: Its described and natural outbreak and an experimental poisoning. In the outbreak, three bovines in the acute phase and three in the chronic phase were examined. Blood samples were collected from all of these animals in order to measure serum levels of aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), urea, and creatinine. The first three animals underwent necropsy and histopathological evaluation. The experiment was conducted with two nine-month-old calves that received an oral paste made with crushed S. fissuratum fruits mixed with water. These fruits were collected at a farm at which cattle poisoning cases had occurred. Blood samples were collected in order to measure serum levels of AST, GGT, urea, and creatinine, before plant administration and then daily during the experimental period. Skin biopsies were taken before plant administration and new one after the first signs of skin lesions. The natural outbreak affected 52 of 160 bovine (31 calves and 21 cows) in the lot. Two calves and 14 cows died. Clinical signs consisted of depression, ataxia, incoordination, behavioral changes, decubitus, and death. One animal that died and 36 others that recovered had photodermatitis. Necropsy findings in the animals consisted of bad corporal condition, pale kidneys, evidence of liver lobular pattern, dry rumen contents, and full bladder. In two animals, fruit seeds were found in the rumen, and one animal had ulcers and transmural edema in the abomasum. Microscopically, mild to moderate renal tubular distension, accumulation of proteinaceous material in lumen with mild to moderate swelling, and epithelial necrosis. In the liver, swelling of hepatocytes and moderate bile stasis was detected. Enzymes values in all evaluated bovines were higher than those considered normal for the species. Experimentally, both calves became ill and one died. The clinical signs were apathy, inappetence, wobbling, weight loss, and goosebumps. One of them had jaundice, tearing, photophobia, ear skin detachment, and ulcers at the muzzle, nostrils and ventral face of the tongue. This animal was euthanized in extremis, and the necropsy findings showed generalized jaundice, evidence of increased liver lobular pattern, thick bile, pale kidneys, and esophageal, tongue, and epiglottal ulcers. Microscopically, the lesions were similar to those described during the natural outbreak. The skin biopsy from the calf that recovered showed perivascular edema and mild eosinophilia.Discussion: The diagnosis was made based on clinical signs, necropsy findings, histopathological lesions, and epidemiological analysis. Experimentally, the plant was toxic at the administered doses. Photosensitization was the most common clinical sign during the natural outbreak and until now, has never been experimentally reproduced. Based on histopathological lesions observed in this study, we can consider that is from hepatogenous origin. The results showed that the kidney lesions have an important role during the pathogenesis caused by this poisoning and during disease evolution.

Epithelial TSC1-mTOR controls RIPK3-dependent necroptosis and susceptibility to IBD by sensing dietary and microbial metabolites

Abstract Intestinal epithelial cells (IECs) are responsible for the sensing and uptake of dietary and microbial metabolites, yet how dietary and microbial components exert on intestinal homeostasis and Inflammatory Bowel Diseases (IBDs) has yet to be better understood. Here we firstly identified epithelial necroptosis as a key process influenced by nutrients protein and fatty acid, as feeding high-protein or high-fat diet to mice led to mTOR activation and aberrant necrosis in IECs. Notably, genetic ablation of TSC1 in the IECs mimicked the effect of high-protein/fat feeding, resulting in mTOR dysregulation, epithelial necrosis and hyper-susceptibility to colitis. Mechanistically, our data demonstrated that aberrant activation of mTOR could result in the overexpression of RIPK3 in enterocytes, whereby rendering epithelial necrosis and ensuing colitis, a process could be reversed by microbiota-depletion or rapamycin treatment. Remarkably, this mTOR-RIP3-necrosis axis seemed to underlie the pathogenesis of human IBDs as well. Furthermore, we found that mTOR-regulated selective autophagy might have contributed to the degradation of RIPK3 in IECs, revealing an intricate association between autophagy and necroptosis. Together, our data unravel an unsuspected link between metabolism and necroptosis, and highlight a crucial role for TSC1-mTOR in the maintenance of intestinal homeostasis and prevention of colonic inflammation through coordinating epithelial metabolism, autophagy and necroptosis.

Uremic encephalopathy in a rhesus macaque (Macaca mulatta): A case report and a brief review of the veterinary literature.

Uremic encephalopathy is uncommon yet is one of the most severe complications of renal failure. We present a case of acute renal failure and associated cerebral and vascular lesions consistent with uremic encephalopathy in a rhesus macaque (Macaca mulatta). A 14-year-old, female, specific-pathogen-free rhesus macaque presented in lateral recumbency, obtunded, severely dehydrated, and hypothermic, with severe azotemia, mild hyponatremia, hypokalemia, hypochloremia, increased anion gap, and hypercholesterolemia. Due to poor prognosis, the animal was euthanized and a complete necropsy was conducted. The animal had diffuse proximal renal tubular epithelial necrosis and loss; regeneration of tubular epithelium was not observed. There was bilateral necrosis and loss of neurons and glial cells in the hippocampus and deep cerebral cortex with edema and multifocal areas of hemorrhage. We present the first reported case of uremic encephalopathy in a rhesus macaque and describe the associated cerebral and vascular lesions.

Mitochondrial DNA induces Foley catheter related bladder inflammation via Toll-like receptor 9 activation

Bladder instrumentation engages the innate immune system via neutrophil activation, promoting inflammation and pain. Elevated levels of mitochondrial DNA (mtDNA) have been associated with tissue damage and organ dysfunction. We hypothesized that local bladder trauma induced by a Foley catheter (FC) will result in mtDNA release, migration of neutrophils into the bladder lumen, and activation of the Toll-like receptor 9 (TLR9) and nuclear factor kappa B (NF-κB) pathway leading to bladder tissue damage. We randomized 10 swine into two groups receiving uncoated, or chloroquine/N-Acetylcysteine (CQ/NAC)-coated FCs. Urine samples were analyzed for mtDNA activation of TLR9/NF-κB as demonstrated by indicators of neutrophil adhesion, migration, and activation. We found that uncoated FCs resulted in a unique active neutrophil phenotype that correlated with bladder epithelial injury, neutrophilia, necrosis, mtDNA release, TLR9/NF-κB activation, transcription and secretion of pro-inflammatory cytokines, and enhanced respiratory burst. In our study we observed that the high levels of mtDNA and elevated TLR9/NF-κB activity were ameliorated in the CQ/NAC-coated FC group. These findings suggest that post-migrated bladder luminal neutrophils are involved in local tissue damage and amelioration of the mtDNA/TLR9/NF-κB inflammatory axis may represent a therapeutic target to prevent inflammation, and bladder tissue injury.

N-acetylcysteine transforms necrosis into apoptosis and affords tailored protection from cisplatin cytotoxicity

Nephrotoxicity is the main limitation to the dosage and anticancer efficacy of cisplatin. Cisplatin produces tubular epithelial cell apoptosis and necrosis depending on the concentration of the drug. Protection from cisplatin nephrotoxicity must therefore tackle both cell death modes. For its ability to reduce cisplatin reactivity, in addition to its antioxidant effect, we tested and found that N-acetylcysteine (NAC) was most effective at inhibiting cisplatin cytotoxicity. NAC has no significant effect on cell death induced by either cycloheximide or Fas activation, indicating a rather selective action. Pt-DNA-binding experiments suggest that the differential effectiveness of NAC is due to its capacity to quench cisplatin reactivity inside the cell. NAC abolishes cisplatin-induced apoptosis, and transforms the necrosis induced by high concentrations of cisplatin into apoptosis. In fact, NAC allows the anti-apoptotic molecule Bcl-2 to reduce the cell death caused by pro-necrotic concentrations of cisplatin, to a significantly greater extent than in the absence of NAC. In rats, a dosage of NAC that significantly ameliorates cisplatin nephrotoxicity, has little effect on gentamicin nephrotoxicity. These characteristics provide NAC with a rationale as a potential nephroprotectant specifically tailored to and especially effective for therapeutic courses with platinated antineoplastics, which prompts to deepening into further preclinical knowledge, and to initiate clinical studies with NAC and mixed therapies composed of NAC and antiapoptotic drugs.

Ecotoxicological impacts of exposure to copper oxide nanoparticles on the gill of the Swan mussel, Anodonta cygnea (Linnaeus, 1758)

ABSTRACTThis study was conducted to investigate the ecotoxicological effects of exposure to copper oxide nanoparticles (CuO NPs) on the gill of the swan mussel Anodonta cygnea using several approaches including qualitative and quantitative histopathology, ultra-morphology (scanning electron microscopy [SEM]) and measures of clearance rate (CR) and bioaccumulation of CuO NPs. Histological alterations in mussels exposed to 0.25 (T1), 2.5 (T2) and 25.0 µg L−1 (T3) CuO NPs for 12 days include changes in the length and form of gill lamellae, changes in inter-lamellar spaces, epithelial hyperplasia, atrophy and tissue rupture. Ultra-morphological changes following CuO NP exposure included epithelial hyperplasia and hypertrophy, epithelial lifting, tissue rupture (water channel fusion) and extensive necrosis of the gill surfaces. IGill (gill damage severity) index values for both histopathological and ultra-morphological data were significantly (P < 0.05) higher in T3. The CR of mussels was significantly (P < 0.05) lower in the highest exposure concentration (CR = 39.21 ± 16.85 [L min−1 g−1 dry weight]) in comparison to controls (CR = 108 ± 47.14 [L min−1 g−1 dry weight]). CuO NPs accumulated in exposed mussels at all exposure concentrations until day 4, but there was no further change in accumulation levels by the end of the exposure period. The accumulated content of CuO NPs was significantly (P < 0.05) lower in 25.0 µg L−1 exposure concentration. Based on these results, significant accumulation of CuO NPs in the gills of swan mussel could affect histological and ultra-structural characteristics of this organ and consequently have deleterious impacts on its filtration activity.

Effect of tadalafil on chronic pelvic pain and prostatic inflammation in a rat model of experimental autoimmune prostatitis.

Experimental autoimmune prostatitis (EAP) shares important clinical features with clinical chronic prostatitis/chronic pelvic pain. We investigated the effect of tadalafil on pelvic pain and prostatic inflammation in a rat EAP model. EAP was induced in rats by intradermal injection of rat prostate antigen and complete Freund's adjuvant on days 0 and 28. Rats were treated with tadalafil (2 mg/kg, p.o.; EAP-tadalafil) or vehicle (EAP-vehicle) once daily from day 0, while sham-operated animals were treated with vehicle only (Sham). Tactile allodynia was measured on days 28, 35, and 42 by applying von Frey filaments to the lower abdomen as an index of pelvic pain. On day 42, the plasma immunoglobulin G (IgG) concentration and the testosterone/estradiol ratio were measured and histopathological analysis of the prostate was performed. Tactile allodynia in the pelvic region was observed on days 28, 35, and 42 after EAP induction. The tactile allodynia observed on day 42 was significantly reduced by repeated treatment with tadalafil. Plasma IgG concentrations increased after EAP induction but the increase was not changed by tadalafil treatment. Prostate tissues were characterized by epithelial necrosis, infiltration of neutrophils and/or lymphocytes to acini and stroma, and fibrosis, in addition to a high stroma-to-epithelium ratio. Tadalafil treatment significantly suppressed the severity of the lesions. EAP rats developed pelvic pain, prostatic inflammation and increased plasma IgG concentrations. Tadalafil inhibited the chronic pelvic pain and prostatic inflammation, suggesting that its anti-inflammatory action may contribute to its blocking of pain development in the EAP model.

Severe Intestinal Inflammation in the Small Intestine of Mice Induced by Controllable Deletion of Claudin-7.

As a potential tumor suppressor gene, Claudin-7 (Cldn7), which is a component of tight junctions, may play an important role in colorectal cancer occurrence and development. To generate a knockout mouse model of inducible conditional Cldn7 in the intestine and analyze the phenotype of the mice after induction with tamoxifen. We constructed Cldn7-flox transgenic mice and crossed them with Villin-CreERT2 mice. The Cldn7 inducible conditional knockout mice appeared normal and were well developed at birth. We induced Cldn7 gene deletion by injecting different dosages of tamoxifen into the mice and then conducted a further phenotypic analysis. After induction for 5days in succession at a dose of 200µl tamoxifen in sunflower oil at 10mg/ml per mouse every time, the mice appeared dehydrated, had a lower temperature, and displayed inactivity or death. The results of hematoxylin-eosin staining showed that the intestines of the Cldn7 inducible conditional knockout mice had severe intestinal defects that included epithelial cell sloughing, necrosis, inflammation and hyperplasia. Owing to the death of ICKO mice, we adjusted the dose of tamoxifen to a dose of 100µl in sunflower oil at 10mg/ml per mouse (aged more than 8weeks old) every 4days. And we could induce atypical hyperplasia and adenoma in the intestine. Immunofluorescent staining indicated that the intestinal epithelial structure was destroyed. Electron microscopy experimental analysis indicated that the intercellular gap along the basolateral membrane of Cldn7 inducible conditional knockout mice in the intestine was increased and that contact between the cells and matrix was loosened. We generated a model of intestinal Cldn7 inducible conditional knockout mice. Intestinal Cldn7 deletion induced by tamoxifen initiated inflammation and hyperplasia in mice.

Nonclinical Safety Assessment of the γ-Secretase Inhibitor Avagacestat

The toxicity of avagacestat, a sulfonamide-based gamma (γ)-secretase inhibitor that was in development as a treatment for Alzheimer's disease, was evaluated in a comprehensive nonclinical toxicology program that included 6-month and 1-year repeat-dose toxicity studies in rats and dogs, respectively. There was a spectrum of mechanism-based changes attributed to inhibition of Notch signaling that regulates the differentiation and proliferation of cells throughout development and in adult tissues. In both rats and dogs, ovarian follicular degeneration and atrophy and a low incidence of granulosa cell hyperplasia and benign granulosa-thecal cell tumors were observed. Gastrointestinal (GI) findings, including goblet cell metaplasia, dilatation of intestinal crypts/glands, mucosal epithelial necrosis and regeneration, and villous atrophy, were limited to dogs that had clinical evidence of GI toxicity. Other avagacestat-related findings attributed to interference with Notch signaling included decreases in peripheral lymphocytes (T and/or B cells) and lymphoid depletion in lymph nodes and the spleen in both species, as well as epiphyseal cartilage and trabecular bone changes in rats. Pharmacologically mediated decreases in brain and cerebrospinal fluid levels of β-amyloid (Aβ) peptides Aβ40 and Aβ42 and decreased expression of white blood cell mRNA levels of the Notch-regulated gene hairy and enhancer of split-1 confirmed target engagement at all doses. Reductions in brain Aβ peptide levels (22 to 34%) in dogs after 1 year at exposures up to the no-observed-effect level for GI toxicity of 1.1× the human plasma exposure, and reversible GI changes at a 3.2× multiple, indicated that a sustained pharmacodynamic effect was attained at exposures without dose-limiting toxicity.

Targeting intestinal epithelial cell–programmed necrosis alleviates tissue injury after intestinal ischemia/reperfusion in rats

BackgroundIntestinal dysfunction, especially acute pathologies linked to intestinal ischemia/reperfusion (I/R) injury, is profoundly affected by inflammation and improper execution of cell death. Few studies have examined the efficacy of combined strategies in regulated intestinal epithelial necrosis after intestinal I/R. Here, we evaluated the functional interaction between poly (adenosine diphosphate-ribose) polymerase 1 (PARP-1)-induced parthanatos and receptor-interacting protein 1/3 (RIP1/3) kinase-induced necroptosis in the pathophysiological course of acute ischemic intestinal injury. MethodsAnesthetized adult male Sprague–Dawley rats were subjected to superior mesenteric artery occlusion consisting of 1.5 h of ischemia and 6 h of reperfusion. The PARP-1-specific inhibitor PJ34 (10 mg/kg) and the RIP1-specific inhibitor Necrostatin-1 (1 mg/kg) were intraperitoneally administered 30 min before the induction of ischemia. ResultsIntestinal I/R was found to result in PARP-1 activation and RIP1/3-mediated necrosome formation. PJ34 or Necrostatin-1 treatment significantly improved the mucosal injury, while the combined inhibition of PARP-1 and RIP1/3 conferred optimal protection of the intestine. Meanwhile, results from terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay showed a decrease in intestinal epithelial cell death. Interestingly, we further showed that PARP-1 might act as a downstream signaling molecule of RIP1 in the process of I/R-induced intestinal injury and that the RIP1/PARP-1-dependent cell death signaling pathway functioned independently of caspase 3 inhibition. ConclusionsThe results of our study provide a molecular basis for combination therapy that targets both pathways of regulated necrosis (parthanatos and necroptosis), to treat acute intestinal I/R-induced intestinal epithelial barrier disruption.