Hepatocyte Swelling Research Articles

P1.14-51 LOC440416 lncRNA-Mediated Cell Cycle Arrest Regulated Crizotinib-Induced Hepatotoxicity

To evaluate crizotinib-induced hepatotoxicity in mice and investigate the role of cell cycle arrest mediated by LOC440416lncRNA during liver injury, furthermore, to provide experiment evidence for screening available hepar protector and expending the clinical application of crizotinib. The levels of blood serum ALT, AST, LDH were detected using ICR mouse model. Liver samples of mouse fixed were stained with hematoxylin and eosin for histopathological analysis. The cell inhibition rate of primary hepatocytes was detected by MTT assay.Cell cyclin distribution was detected by flow cytometry(FCM) and the expressions of cell cycle protein cyclinB1 and CDK1 were detected with Westblot after mouse primary hepatocytes were treated with 2.5 μM, 5μM, 10 μM crizotinib for 48h. Apoptosis was evaluated by DAPI staining and PI staining for flow cytometry and the expressions of cell apoptosis protein c-PARP and c-caspase-3 were detected with Westblot. LncRNAs regulating hepar cell cycle arrest were screened using Microarray Chip technique after primary hepatocytes were treated with 5μM crizotinib for 24h. The expressions of cell cyclin protein cyclin B1 and CDK1 were tested with Westblot, after siRNA gene silencing technology was used to knock down LOC440416lncRNA. The levels of blood serum ALT, AST, LDH elevated significantly in mice after crizotinib treatment. Slices of liver by HE staining for analysis showed cellular swelling of hepatocytes, nucleolus staining deepened and karyokinesis phenomenon with inflammatory cell infiltrate. In vitro, MTT assay showed crizotinib-induced hepatotoxicity in mouse primary hepatocytes presented dose-dependented and IC50 value was 4.6μM. DAPI staining exhibited clear chromatin condensation, fragment and apoptotic bodies and FCM showed sub-G1 apoptosis peak was formed after 5μM and10 μM crizotinib treatment. Westblot demonstrated caspase-3 activation and PARP cleavage. All these data revealed that crizotinib-induced hepatotoxicity was through the process of apoptosis. FCM showed the population of G2/M phase elevated, these of G0/G1 phase decreased and G2/M arrested while the concentration of crizotinib was increased. Reduced protein levels of Cyclin B1 and increased protein level of CDK1 were also observed with Westblot. After crizotinib treatment, sixteen elevated or decreased lncRNAs were found more than ten times separately by terms of Microarray Chip technique. LOC440416lncRNA regulating cell cycle arrest was screened by Real-time PCR. The restoration of reduced Cyclin B1, increased CDK1 expressions, even if hepatocytes were treated with crizotinib, after siRNA gene silencing technology was used to knock down LOC440416lncRNA. Crizotinib-induced hepatotoxicity was demonstrated by experiment in vivo and in vitro. Cell cyclin G2/M check point arrest accompanied with decreased expression of cyclin B1 protein and increased expression CDK1, mediated by LOC440416 lncRNA, could be the possible mechanism.

The Potential of Neoagaro-Oligosaccharides as a Treatment of Type II Diabetes in Mice.

Type 2 diabetes mellitus (T2DM) accounts for more than 90% of cases of diabetes mellitus, which is harmful to human health. Herein, neoagaro-oligosaccharides (NAOs) were prepared and their potential as a treatment of T2DM was evaluated in KunMing (KM) mice. Specifically, a T2DM mice model was established by the combination of a high-fat diet (HFD) and alloxan injection. Consequently, the mice were given different doses of NAOs (100, 200, or 400 mg/kg) and the differences among groups of mice were recorded. As a result of the NAOs treatment, the fasting blood glucose (FBG) was lowered and the glucose tolerance was improved as compared with the model group. As indicated by the immunohistochemistry assay, the NAOs treatment was able to ameliorate hepatic macrovesicular steatosis and hepatocyte swelling, while it also recovered the number of pancreatic β-cells. Additionally, NAOs administration benefited the antioxidative capacity in mice as evidenced by the upregulation of both glutathione peroxidase and superoxide dismutase activity and the significant reduction of the malondialdehyde concentration. Furthermore, NAOs, as presented by Western blotting, increased the expression of p-ERK1/2, p-JNK, NQO1, HO-1, and PPARγ, via the MAPK, Nrf2, and PPARγ signaling pathways, respectively. In conclusion, NAOs can be used to treat some complications caused by T2DM, and are beneficial in controlling the level of blood glucose and ameliorating the damage of the liver and pancreatic islands.

Effects of replacing fishmeal protein with poultry by-product meal protein and soybean meal protein on growth, feed intake, feed utilization, gut and liver histology of hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) juveniles

Two consecutive eight-week growth trials were undertaken to evaluate the effects of replacing fishmeal protein (FMP) with poultry by-product meal protein (PBMP) and soybean meal protein (SBMP) on growth, feed intake, feed utilization, gut and liver histology of hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) juveniles. In trial 1, eight isoenergetic (340 kcal per 100 g dry matter) and isoproteic (53.5% of dry matter) experimental diets were formulated to replace 0%, 10%, 20%, 30%, 40%, 50%, 60% and 70% FMP with PBMP, being abbreviated as FMP, PBMP10, PBMP20, PBMP30, PBMP40, PBMP50, PBMP60 and PBMP70, respectively. Based on the results of trial 1, another six isoenergetic (340 kcal per 100 g dry matter) and isoproteic (53.5% of dry matter) experimental diets including a high FM (86.64%) diet (FMP) and five low-FM (25.99%) diets were formulated in trial 2, and in the low-FM diets, 0%, 7%, 14%, 21% and 21% PBMP was replaced with SBMP, being abbreviated as PBMP, SBMP7, SBMP14, SBMP21, SBMP28, respectively. The initial average body weights of experimental fish were 6.0 ± 0.05 g in trial 1 and 6.8 ± 0.08 g in trial 2 (means ± S.D.), and fish were fed their described diets by hand to apparent satiation twice daily (08:00 and 16:30) at a density of 30 fish/cage (trial 1) or 15 fish/cage (trial 2). Experimental cages were labeled and located in connective 6-m3 indoor concrete tanks (L 3 m × W 2 m × H 1 m) with 3 cages occurring in each tank. Each treatment had three replicates both in trial 1 and in trial 2.Results showed that in trial 1, weight gain% (WG%), daily feed intake (DFI), feed conversion ratio (FCR), protein efficiency ratio (PER) as well as protein productive value (PPV) were not significantly affected by different FMP replacements with PBMP. Fish fed FMP, PBMP10 and PBMP20 had higher hepatosomatic index (HSI) and condition factor (CF) than fish fed PBMP30, PBMP40, PBMP50, PBMP60 and PBMP70, and intraperitoneal fat (IPF) ratio was increased as dietary PBM inclusion levels increased. Whole-body and muscle lipid contents of fish were elevated by the increments in PBM. High FMP replacements (50%–70%) by PBMP induced steatosis in hepatocytes. Different replacements of FMP by PBMP had no significant effects on gut morphology of experimental fish. In trial 2, there were also no remarkable variations in WG%, DFI, FCR and PER among various experimental treatments. Fish fed SBMP7, SBMP14, SBMP21 and SBMP28 showed similar fold height (hF), enterocyte height (hE) as well as microvilli height (hMV) in foregut and hF, hE in midgut in comparison to fish fed PBMP or the high FM, but for midgut hMV and hindgut hF, hE, hMV, fish fed the low-FM diets (PBMP, SBMP7, SBMP14, SBMP21 and SBMP28) displayed lower values of these parameters than fish fed the high-FM diet. Swelling in hepatocytes was observed in fish fed SBMP14, SBMP21 and SBMP28. Generally, replacing 70% FM protein or reducing the FM inclusion level to about 26% with PBMP or the combination of PBMP and SBMP at the 53.5% dietary crude protein level did not negatively affect growth, feed intake as well as feed utilization of hybrid grouper, but liver health and gut morphology of fish fed low FM diets need to be further focused on.

Acute and sub-acute toxicity study of the root extracts of Fagaropsis hildebrandtii in mice and evaluation of their antimicrobial effects

Background:Among the Kamba community of Kenya, roots ofFagaropsis hildebrandtii(FH) are boiled and used in managing cough, fertility problems, and microbial infections. The safety of this plant in oral administration and the validity of the ethnomedical claims is unverified. This study evaluated the toxicity of the aqueous and hexane root extracts of FH in mice and antimicrobial effects againstStaphylococcus aureus,Salmonella typhimuriumandCandida albicans.Methods: Doses (300 and 2000mg/kg) of the extracts were administered orally to mice for 14 days. The weight, feed, and water consumption, organ weight of mice and gross macroscopy of the liver were used in evaluating acute toxicity. Mice were additionally treated with 250, 500, and 1000mg/kg body weight doses of the extracts for 28 days and haematological, biochemical, and histological parameters noted. The minimum inhibitory and minimum bactericidal/fungicidal concentrations (MIC; MBC/MFC) of the extracts against the aforementioned pathogens were determined by broth dilution.Results:Acute oral toxicity of the extracts was >2000mg/kg, there were dose dependent changes in haematological and biochemical parameters, all female mice died when treated with doses of 1000mg/kg and doses ≥500mg/kg caused tubular degeneration and haemorrhage of the kidney, cloudy swelling of hepatocytes, and multifocal necrosis and pyknosis in the liver. The MBC/MIC ratio of each of the extracts againstStaph. aureusandS. typhimuriumwas 2, whileC. albicanswas not sensitive to any of the extracts.Conclusions:Long term use of root extracts of FH was associated with dose-dependent changes in the kidney and liver of mice and changes in biochemical and haematological parameters. Root extracts of FH are bactericidal againstStaph. aureusandS. typhimuriumbut have no effect onC. albicans. Future work should aim at identifying the metabolites responsible for the observed toxic and bactericidal effects of the roots of FH.

Toxicological findings about an anticancer fraction with casearins described by traditional and alternative techniques as support to the Brazilian Unified Health System (SUS)

Ethnopharmacological relevanceExtracts, essential oils and molecules from Casearia sylvestris have popularly shown pharmacological actions against chronic diseases, as anxiety, inflammation, cancer and dyslipidemia. In the context of antitumoral therapy, we investigated in vitro, ex vivo and in vivo toxicological changes induced by a Fraction with Casearins (FC) and its component Casearin X isolated from C. sylvestris on animal and vegetal cells, and upon invertebrates and mammals. Material and methodsCytotoxicity was carried out using normal lines and absorbance and flow cytometry techniques, Artemia salina nauplii, Danio rerio embryos and meristematic cells from Allium cepa roots. Acute and 30 days-mice analysis were done by behavioral, hematological and histological investigations and DNA/chromosomal damages detected by alkaline Cometa and micronucleus assays. ResultsFC was cytotoxic against lung and fibroblasts cells and caused DNA breaks, loss of integrity and mitochondrial depolarization on ex vivo human leukocytes. It revealed 24 h-LC50 values of 48.8 and 36.7 μg/mL on A. salina nauplii and D. rerio embryos, reduced mitotic index of A. cepa roots, leading to cell cycle arrest at metaphase and anaphase and micronuclei. FC showed i.p. and oral LD50 values of 80.9 and 267.1 mg/kg body weight. Subacute i.p. injections induced loss of weight, swelling of hepatocytes and tubules, tubular and glomerular hemorrhage, microvesicular steatosis, lung inflammatory infiltration, augment of GPT, decrease of albumin, alkaline phosphatase, glucose, erythrocytes, and lymphocytes, and neutrophilia (p > 0.05). FC-treated animals at 10 mg/kg/day i.p. caused micronuclei in bone marrow and DNA strand breaks in peripheral leukocytes. ConclusionsThis research postulated suggestive side effects after use of FC-related drugs, demonstrating FC as antiproliferative and genotoxic on mammal and meristematic cells, including human leukocytes, teratogenicity upon zebrafish embryos, myelosuppression, clastogenicity, and morphological and biochemical markers indicating liver as main target for FC-induced systemic toxicity.

Histopathological changes in placenta and liver of pregnant rats administered with aqueous extract of Dioscorea hispida var. daemona (Roxb) Prain & Burkill

Dioscorea hispida var. daemona (Roxb) Prain & Burkill (DH), also known a tropical yam or intoxicating yam is a bitter wild tuber which is consumed as a staple food and traditionally used as a remedy in Malaysia. However, DH is also notorious for its intoxicating effects and there is currently a dearth of study of possible effects of DH on liver and placental tissues and hence its safe consumption warrants in-depth investigation. This study was therefore designed to investigate into the effect of DH on liver and placenta of pregnant rat via histopathological examination. Thirty pregnant Sprague-Dawley rats were randomly divided into five groups consisting of a control (distilled water) and four DH aqueous extract groups (250, 500, 1000 and 2000 mg/kg body weight). The extracts were administered via oral gavage daily throughout the study and animals were sacrificed on day 21. Paraffin-embedded, hematoxylin and eosin stained sections of placenta and liver were examined. Significant changes (p < 0.05) were observed on relative liver and placental weights of animals treated with 2000 mg/kg body weight DH extract. The placental numbers were decreased with the increased of DH extract concentration. Liver histological examination in all treated groups showed that tissues underwent degeneration characterized by hepatocyte swelling, cytoplasmic vacuolation, cytolysis, margination and clumping of nucleus chromatin. Changes of the basal and labyrinth zone were observed in placental tissues in all treated groups. Glycogen cells were reduced with fibrin deposition in the basal zone, while irregular vessel formation was demonstrated in the labyrinth zone. UHPLC-ESI-MS analysis showed the presence four steroidal saponins DH. In conclusion, DH aqueous extract exert hepatotoxicity and adverse effects on the placenta of rats. However, the underlying mechanism and phytochemicals inducing the observed toxicity require further investigation.

Influence of ferulic acid consumption in ameliorating the cadmium-induced liver and renal oxidative damage in rats.

The aim of this study relates to the modulatory role of ferulic acid (FA) against cadmium (Cd)-induced oxidative stress in the liver and kidney of male Wistar albino rats. Cd is an extremely toxic industrial and environmental pollutant and is well known for its varied toxic clinical manifestations. FA is a derivative of curcumin and a ubiquitous phenolic compound having a wide range of therapeutic activities. In the current study, Cd (10mg/kg) was administered subcutaneously for 15 and 30days to induce hepato-renal toxicity. Cd concentration was found to be significantly high in Cd-intoxicated rats (liver > kidney) while the supplementation of FA (50mg/kg) significantly reduces the Cd concentration in liver and kidney tissues. Reduced body and organ weights and food and water consumption and increased rectal temperature were noticed in Cd-treated rats while these parameters were significantly ameliorated in FA-supplemented rats. Liver and kidney damage induced by Cd was significantly revealed by the reduction in serum total protein contents (TPC) and increased activities of serum nitric oxide (NO) levels and hepato-nephrotoxicity marker enzymes, namely aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactic dehydrogenase (LDH), AST:ALT ratio, uric acid, urea, urea nitrogen, and creatinine, along with the increased levels of hepatic and renal oxidative stress markers, namely lipid peroxidation (MDA levels), lipid hydroperoxides (LOOH), protein carbonyl content (PCC), total oxidant status (TOS), and oxidative stress index (OSI) in liver and kidney tissues. In addition, the toxicity of Cd was also evidenced by a significant decrease in the levels of total thiols (TTH), total antioxidant concentration (TAC), enzymatic antioxidants (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)), and non-enzymatic antioxidants (reduced glutathione (GSH) and total free sulfhydryl groups (TSH)). Administration of FA significantly restored the serum total protein levels and activities of serum NO levels and hepatic and renal marker enzymes to normal levels in comparison with Cd-intoxicated rats. Furthermore, FA significantly reduced the oxidative stress markers and recuperated the levels of antioxidant defense in the liver and kidney as evidenced by native PAGE and spectrophotometric assays, correlation and regression analysis and multivariate analysis of variance (MANOVA), and inferring the antioxidant role of FA. Histopathological damage due to Cd intoxication in the liver and kidney is demonstrated as vasodilatation and congestion in central veins and sinusoids as well as around the glomerulus, infiltration of mixed inflammatory cells and peripheral hemorrhage, hemorrhagic and enlarged sinusoids, disorganization of the hepatic parenchyma, focal necrosis, swelling of hepatocytes, calcified tissue inside blood vessels, hepatocyte degeneration and vacuolization of liver cells, hyaline casts, degenerated glomerulus with wide space and detached basement membrane, distal tubule with wide lumen, deformed proximal tubules with detached brush border, and degeneration and hyalinization of glomerular tuft. But, FA significantly reduced the toxicity of Cd and protected the normal histological architecture of the liver and kidney tissues. Cd-intoxicated rats were associated with a significant upregulation of TNF-α, COX-2, and HSP70 proteins, whereas treatment with FA caused downregulation of the above inflammatory markers indicating the anti-inflammatory role of FA. Principal component analysis (PCA) and Euclidean similarity measure studies clearly indicate that the liver is more prone to Cd toxicity than the kidney and FA supplementation significantly prevents oxidative stress, augmenting antioxidative status, and regaining histological parameters of the liver and kidney to normal, indicating hepato-nephroprotective, antiradical, antioxidant, and anti-inflammatory effects of this phenolic compound.

Serum ferritin in neonatal cholestasis: A specific and active molecule or a non-specific bystander marker?

BackgroundSerum ferritin (SF) and consequently hepatic iron have long been considered important in liver fibrosis progression. They have been studied in different liver diseases with no previous reports in neonatal cholestasis (NC). This study aimed to measure SF in different etiologies of NC and investigate its relation to hepatic iron and fibrosis. MethodsSF was measured in 75 infants, including 50 with NC and 25 with sepsis. SF was compared between these two groups. Biochemical parameters, hepatic iron grades, and liver fibrosis and other histopathological characteristics and correlated with SF were assessed in NC group. Finally, a comparison between intrahepatic cholestasis and obstructive etiology was performed. ResultsSF was elevated in NC (1598 ± 2405 ng/mL) with no significant difference from those with sepsis (P = 0.445). NC and sepsis constituted augmenting factors leading to more elevation of SF (2589 ± 3511 ng/mL). SF was significantly correlated with hepatic iron grades (r = 0.536, P < 0.0001) and a cut-off value of 803.5 ng/mL can predict higher grades (≥ grade 3) of iron deposition with sensitivity of 100%, specificity of 70% and accuracy of 85%. Moreover, SF was significantly higher (P < 0.0001) in those with intrahepatic cholestasis (2602 ± 3154 ng/mL) and their prevalent pathological findings of giant cell transformation (P = 0.009) and hepatocyte swelling (P = 0.023) than those with obstructive etiology (672 ± 566 ng/mL) and their prevalent pathological findings of ductular proliferation (P = 0.003) and bile plugs (P = 0.002). SF was unrelated to the grade of liver fibrosis (P = 0.058). ConclusionsSF is non-specifically elevated in NC, with positive correlation to hepatic iron grades. SF ≥ 803.5 ng/mL can predict higher grades (≥ grade 3) of hepatic iron. However, an active role of increased SF and hepatic iron in disease progression remains questionable.

Characterization and Pathology of Aeromonas veronii Biovar Sobria from Diseased Sheatfish Silurus glanis in China

Aeromonas spp., is an important opportunistic pathogen responsible for great economic losses in aquaculture. In this paper, we report our study of an Aeromonas infection that occurred in cultured sheatfish Silurus glanis in May 2016 in a farm in Tianjin city. Typical external symptoms included petechial skin hemorrhages, ascitic distension of the abdomen, redness and swelling of the anus. The isolate SAV-165 from the diseased fish was identified as A. veronii biovar sobria by biochemical properties and molecular techniques. Experimental challenge showed that a 50% lethal dose (LD50) of SAV-165 was 1.59 × 106 CFU/ml in zebrafish Danio rerio. The antimicrobial susceptibility test indicated that the isolate was susceptible to aminoglycosides and chloramphenicol, but resistant to penicillins and sulphonamides. Histopathological examination showed fatty changes and swelling of hepatocytes in the liver, congestion in the submucosa of intestine and the renal veins of the kidney. This is the first report of A. veronii biovar sobria infection in S. glanis in China.

The protective effect of Protectin DX on acute liver injury induced by sepsis in mice and the underlying mechanism

Objective To explore the effect of Protectin DX(PDX) on acute liver injury(ALI) induced by sepsis in mice and the underlying mechanism. Methods Mice received cecum ligation and puncture(CLP) to induce sepsis-associated acute liver injury. Male C57BL/6 mice were randomly (random number) divided into 3 groups (n=10 each group): (1) sham group (S group), (2) CLP group and (3) CLP + PDX group (PDX group ). Mice in the PDX group were received PDX 1 μg (intraperitoneal injection). One hour after CLP operation, mice in the S and CLP groups were received equal amounts of saline. The serum and liver tissues were collected at 24 h after CLP. The histological changes of the liver were observed by HE staining. The ALT and AST levels in the serum were assessed by using the automatic biochemical analyzer. The levels of cytokines (TNF-α, IL-6, IFN-γ and IL-10) in the serum were quantified by ELISA. MPO activity in the liver tissues were assessed. Western blot was used to detect the expression of pNF-kB p65 and NF-kB p65 in liver tissues. Results Compared with the S group, HE staining in the CLP group showed disordered hepatic cords, hepatocyte swelling and necrosis, infiltrations of inflammatory cells, congestion and bleeding, and the score of liver injury was increased significantly (P<0.05). Levels of ALT, AST, TNF-α, IL-6, IFN-γ, and IL-10 were increased in the CLP group (P<0.05). The activities of NF-κB and MPO in the liver tissues were obviously enhanced (P<0.05). The levels of liver injury, cytokines (TNF-α, IL-6, IFN-γ), MPO and activities of NF-κB in the CLP+PDX group were significantly decreased when compared with those in the CLP group (P<0.05),while the concentration of IL-10 was significantly increased (P<0.05). Conclusions PDX can alleviate sepsis-induced acute liver injury through inhibiting NF-KB activity in the liver tissues. Key words: Protectin DX; Sepsis; Acute liver injury; NF-κB

Experimental poisoning in broiler chickens by Senecio vernonioides, Senecio conyzaefolius and Senecio paulensis

ABSTRACT: Plants of the genus Senecio sp. are known to produce hepatic lesions in different animal species, including man. To evaluate the toxicity of three species of Senecio found in regions where cattle are bred in the state of Santa Catarina, experiments were conducted on broiler chickens (Gallus domesticus). Green leaves of S. conyzaefolius, S. vernonioides and S. paulensis were collected and dried in the shade. After drying, the leaves were ground, mixed into the feed and fed to the chickens divided into 3 groups. Group 1 and Group 2 received single doses of 5g/kg and 20g/kg, respectively. Group 3 was given daily doses of 1g/kg for 20 days (values corresponding to green plant) and Group 4 (control) received free feed of the plant. For each group, five chickens were submitted to necropsy at 30 days and five to 60 days after planting, and five chickens that consumed S. vernonioides and S. conyzaefolius at the dose of 1g/kg for 20 days were necropsied at 90 days after the beginning of the experiment. Macroscopic lesions were observed in chickens that consumed S. vernonioides at a dose of 1 g/kg for 20 days in birds necropsied at 30, 60 and 90 days and were characterized mainly by liver of firm consistency, diminished in size with yellowish coloration and evident lobular pattern, ascites and hydropericardium. In microscopy the main lesions observed were megalocytosis, hepatocyte swelling, fibrosis and biliary hyperplasia and were more intense at the dosage of 1 g/kg for 20 days for S. vernonioides and less severe in the chickens that ingested S. conyzaefolius. The chickens that received S. paulensis showed no macroscopic and microscopic lesions.

Identification of hepatotoxicity and renal dysfunction of pyrene in adult male rats.

Polycyclic aromatic hydrocarbons (PAHs) are a group of persistent organic pollutants primarily formed from the incomplete combustion of carbonaceous materials, and have adverse effects on human health. In this study, we investigated whether pyrene, a PAH consisting of 4 fused benzene rings, has adverse effects on rat. Adult male Sprague-Dawly rats were treated daily by oral gavage with vehicle (corn oil) or pyrene at doses of 375, 750, 1500, or 2200 mg/kg/day for 4 days. The results showed that pyrene caused hepatotoxicity in rats. When compared with the control group, relative liver weights, plasma alanine aminotransferase, and direct bilirubin levels significantly increased after pyrene exposure. Hepatocyte swelling and degeneration and decreased hepatic total glutathione (GSH) levels were also found in pyrene-exposed rats. We further observed that mRNA levels of several hepatic metabolizing enzymes regulated by constitutive androstane receptor (CAR) such as CYP2B1 and CYP2B2 significantly increased in pyrene-exposed rats. These results suggest that decreased GSH levels, elevated hepatic metabolizing enzyme gene expression, and CAR activation are important contributors for pyrene-induced hepatotoxicity in rats. Additionally, we found pyrene significantly induced plasma inflammatory indices including white blood cell and lymphocyte counts. We also observed that pyrene exposure increased relative weight of kidneys and disrupted kidney function with elevated urea and creatinine levels in rats.

Cross-species infection of mice by rabbit hepatitis E virus

Rabbits are recognized as a zoonotic reservoir of hepatitis E virus (HEV) for transmission to humans and other zoonotic reservoirs such as swine. The purpose of this study was to assess the ability of rabbit HEV to cross the species barrier to infect mice and also the usefulness of this animal to study HEV transmission and pathogenesis. In this study, uninfected BALB/c mice were experimentally inoculated with rabbit HEV either via gavage or through contact-exposure with infected mice. Rabbit HEV propagation in mice was evaluated by studying fecal virus shedding, viremia, seroconversion and microscopic liver lesions. Rabbit HEV could be detected in all mice infected by gavage, but only in some contact-exposed mice, with some animals exhibiting fecal virus shedding, seroconversion or viremia (one mouse only). Compared with inoculated mice, anti-rabbit HEV antibody titers and viral copy numbers in fecal and serum samples were lower in contact-exposed mice. Infected mice mainly exhibited phlebitis, hepatocyte swelling and necrosis. Microscopic liver lesion scores for inoculated and contact-exposed infected mice were higher than scores for negative controls. This study therefore demonstrates that rabbit HEV could infect BALB/c mice both though inoculation via gavage and through contact-exposure.

Co-Delivery of Teriflunomide and Methotrexate from Hydroxyapatite Nanoparticles for the Treatment of Rheumatoid Arthritis: In Vitro Characterization, Pharmacodynamic and Biochemical Investigations.

The present investigation was aimed at developing Teriflunomide (TEF) and Methotrexate (MTX) loaded hydroxyapatite nanoparticles and increasing tolerability towards combination therapy against rheumatoid arthritis by reducing hepatotoxicity. Drug-loaded HAp-NPs were synthesized by wet-chemical precipitation method and optimized by Box-Behnken experimental design. The developed NPs were subjected to in vitro and in vivo characterization. In-vivo pharmacodynamics and biochemical studies were performed on adjuvant- induced arthritis model treated with different formulations; MTX-TEF-SOL, TEF-HAp-NP, MTX-HAp-NP, TEF-MTX-HAp-NP, FOLITRAX-10 and AUBAGIO. The size of the optimized formulations, TEF-HAp-NP and MTX-HAp-NP, was found to be 224.3 ± 83.80nm and 268.3 ± 73.86nm with drug loading 53.11 ± 0.84% and 67.04 ± 1.12% respectively. In vitro release of TEF from TEF-HAp-NP (70.41 ± 1.22%) and MTX from MTX-HAp-NP (82.43 ± 1.31%) up to 24h revealed sustained release pattern. Results of the arthritic assessment study showed a significant (P < 0.05) reduction in ankle diameter (61.30 ± 7.42) and arthritis score (2.35 ± 0.24) with a marked restoration of ankle joint micro-architecture in TEF-MTX-HAp-NP treated group. During Hepatotoxicity studies, liver histopathology revealed that the formulation MTX-TEF-HAp-NP was least hepatotoxic with less hepatocyte swelling and fibrous connective tissue proliferation while Folitrax-10 was found to be most hepatotoxic. Biochemical studies revealed that Folitrax-10 significantly (P < 0.05) increased the GOT (313.64 ± 16) and GPT level (334.46 ± 13) while insignificant (P > 0.05) change in GOT (263.68 ± 17) and GPT (229.38 ± 10) level was recorded with TEF-MTX-HAp-NP. We report that the subcutaneous delivery of TEF-MTX-HAp-NP was most effective as it successfully reduced the dosage by half for maximizing therapeutic efficacy and minimizing side effects. Graphical Abstract ᅟ.

Environmentally relevant concentrations of carbamazepine induce liver histopathological changes and a gender-specific response in hepatic proteome of Chinese rare minnows (Gobiocypris rarus)

To assess hepatotoxicity and to determine the underlying mechanisms of carbamazepine (CBZ) toxicity in fish, histopathology and the liver proteome were examined after Chinese rare minnow (Gobiocypris rarus) were exposed to 1, 10, and 100 μg/L CBZ for 28 days. Histopathological changes included disruption of spatial structure, pyknotic nuclei, cellular vacuolization and deformation of cell nuclei, in addition to marked swelling of hepatocytes in all treatment groups. Protein analysis revealed that there were gender-specific responses in rare minnow following exposure, and there were 47 proteins in females and 22 proteins in males identified as differentially abundant following CBZ treatments. Pathway analysis revealed that cellular processes affected by CBZ included apoptosis, cell differentiation, cell proliferation, and the respiratory chain, indicating impaired energy homeostasis. Noteworthy was that 15 proteins identified as different in abundance were associated with carcinogenicity. Relative mRNA levels for select transcripts were consistent with the changes of proteins N-myc downstream regulated gene (NDRG), Tropomyosin 2-Beta (TPM2) and annexin A4 (ANXA4). Protein pyruvate kinase, liver and RBC (PKLR) were increased at 1 and 100 μg/L CBZ without significant difference in transcript levels. These findings characterize molecular responses and histological changes in the liver that generate new insights into CBZ hepatotoxicity in Chinese rare minnow.

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 &amp; 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.

The 1H-NMR-based metabolite profile of acute alcohol consumption: A metabolomics intervention study.

Metabolomics studies of disease conditions related to chronic alcohol consumption provide compelling evidence of several perturbed metabolic pathways underlying the pathophysiology of alcoholism. The objective of the present study was to utilize proton nuclear magnetic resonance (1H-NMR) spectroscopy metabolomics to study the holistic metabolic consequences of acute alcohol consumption in humans. The experimental design was a cross-over intervention study which included a number of substances to be consumed—alcohol, a nicotinamide adenine dinucleotide (NAD) supplement, and a benzoic acid-containing flavoured water vehicle. The experimental subjects—24 healthy, moderate-drinking young men—each provided six hourly-collected urine samples for analysis. Complete data sets were obtained from 20 of the subjects and used for data generation, analysis and interpretation. The results from the NMR approach produced complex spectral data, which could be resolved sufficiently through the application of a combination of univariate and multivariate methods of statistical analysis. The metabolite profiles resulting from acute alcohol consumption indicated that alcohol-induced NAD+ depletion, and the production of an excessive amount of reducing equivalents, greatly perturbed the hepatocyte redox homeostasis, resulting in essentially three major metabolic disturbances—up-regulated lactic acid metabolism, down-regulated purine catabolism and osmoregulation. Of these, the urinary excretion of the osmolyte sorbitol proved to be novel, and suggests hepatocyte swelling due to ethanol influx following acute alcohol consumption. Time-dependent metabolomics investigations, using designed interventions, provide a way of interpreting the variation induced by the different factors of a designed experiment, thereby also giving methodological significance to this study. The outcomes of this approach have the potential to significantly advance our understanding of the serious impact of the pathophysiological perturbations which arise from the consumption of a single, large dose of alcohol—a simulation of a widespread, and mostly naive, social practice.

Assessment of copper nanoparticles (Cu-NPs) and copper (II) oxide (CuO) induced hemato- and hepatotoxicity in Cyprinus carpio

Recently, Cu-based nanoparticles have drawn considerable attention for their various fascinating roles in multiple biological systems. It is recognized that their frequent use can create compatibility challenges for the recipient systems. Nevertheless, it is unclear how various biological interactions affect the compatibility of Cu oxide II (CuO) and Cu oxide nanoparticles (Cu-NPs) for different organisms. Consequently, it has been difficult to perform structured risk assessments for their use in biological systems. Therefore, this study compared the effects of different doses of waterborne Cu-NPs and CuO on the blood and liver of selected groups of Cyprinus (C) carpio. These fish while housed in suitable water tanks were exposed to one of the following treatments for 14 d: control (no added Cu) or 0.5 or 1 or 1.5 mg Cu as Cu-NPs or CuO l–1 of water. We found significant changes in all assessed blood parameters of fish in response to increasing doses from 0 to 1.5 mg of Cu-NPs or CuO. Similarly, increased levels of lipid peroxide and reduced glutathione (GSH) were also observed in the livers of C. carpio in Cu-NPs or CuO treated groups. Enhanced levels of lipid peroxidation and GSH were also recorded in the Cu-NP treated groups compared with the CuO treated groups in a dose dependent manner. The lowest catalase activity was observed in the liver of C. carpio treated with the higer dose of Cu-NPs. Cu-NP or CuO exposure induced significant histological alterations in the liver of C. carpio including focal necrosis, cloudy swelling of hepatocytes, degenerative hepatocytes, vacuolization, pyknotic nuclei, damaged central vein, nuclear hypertrophy, dilated sinusoid, vacuolated degeneration, congestion, and complete degeneration in a dose dependent manner. Substantial alterations in blood and liver specimens were observed in the Cu-NP treated fish when compared with the CuO treated fish. It appeared that the Cu-NPs were more toxic than the CuO as shown by the hemato- and hepatotoxicity in C. carpio of this study.

Effect of Kaempferol Pretreatment on Myocardial Injury in Rats.

The present study was undertaken to evaluate the effect of kaempferol in isoprenaline (ISP)-induced myocardial injury in rats. ISP was administered subcutaneously for two subsequent days to induce myocardial injury. Assessment of myocardial injury was done by estimation of hemodynamic functions, myocardial infarcted area, cardiac injury markers, lipid profile, oxidative stress, pro-inflammatory cytokines and histopathology of heart and liver. Rats pretreated with kaempferol showed reduction in the myocardial infarcted area and heart rate. However, no improvement was observed in change in body weight, mean arterial, systolic and diastolic blood pressure. Kaempferol showed significant decrease in serum LDH, CK-MB, troponin-I and lipid profile. However, highest dose of kaempferol did not reduce the serum triglyceride level. Further, antioxidant enzymes, SOD and catalase, were also higher. However, reduced glutathione, serum SGOT and creatinine did not show any improvement. Kaempferol showed reduction in MDA level. Kaempferol at highest dose showed reduction in pro-MMP-2 expression and MMP-9 level. mRNA expression level of TNF-α was not different in kaempferol-pretreated myocardial injured rats with ISP-alone group. Pretreatment with kaempferol at highest dose showed mild mononuclear infiltration and degenerative changes in heart tissue section of myocardial injured rats. Rats pretreated with kaempferol at higher concentration showed normal cordlike arrangement of hepatocytes with moderate swelling of hepatocytes (vacuolar degeneration) around the central vein. Study suggests that kaempferol attenuated lipid profile, infarcted area and oxidative stress in ISP-induced myocardial injury in rats.

Benzene histotoxic and teratogenic Effects in exposed Mice

Throughout the last two decades, benzene was a prominent source of an occupational and environmental pollution in Iraq. Especially from direct contact with private electric generators which were installed closer to homes or even inside every home in Iraq. As a result of handling without taking enough precautions, environmental threat has increased due to serious toxic, carcinogenic and teratogenic effects of benzene. Toxicity, carcinogenicity and teratogenicity of this chemical carcinogen were experimented in Swiss albino mice by intraperitoneal (i.p.) injection. Six-week-old male mice were used as a model for toxicity and carcinogenicity together with female mice were treated with benzene, and newborns were sacrificed to study the teratogenic impact. During a period of three months, twice per week, male mice were injected with two doses (0.1% and 0.2%) of benzene diluted with corn oil. Female mice were also treated with (0.1% and 0.2%) of benzene on day 7 prior to gestation at 72 hr intervals for one month and sacrificed 10 days after labor. At the end of experiment, sections of different organs were histopathologically observed and significant changes were noticed. Dose-related changes were detected during examination of males' liver showing hepatocyte swelling, degeneration and fibrosis and similarly glomerular enlargement and tubular necrosis of the kidney. New born mice liver sections showed a significant liver and kidney histological changes. These results may explain a correlation between tumor incidence and transplacental benzene exposure in mice and shade the light on the potential health risks that Iraqi community exposed to.Conclusions: When the researchers take into account the degree of damage resulted from direct exposing Albino mice to benzene for a limited period of time, there will be a reasonable wariness because of the increasing level of toxicity and carcinogenicity in the Iraqi environment.