Journal articles: 'Liver and gut histology and morphology' – Grafiati (2024)

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Abstract Background and aims : Background and aims : The liver during fetogenesis does not follow classical lobular pattern. Normal histology of the fetal liver at various stages of development was studied to get insight into the morphology of fetal liver and special function it performs in fetal life. Method: Dissection of 29 normal human fetuses was done and histological findings of liver were noted with respect to the age of fetus. The histology of fetal liver was studied using H & E stain and important features of fetal liver were studied. Result: Fetal liver histology is different from adult liver. Unlike adult liver, fetal liver shows hepatocytes arranged in sheets predominantly with cordlike pattern at places. The sinusoids are dilated and filled with hemopoietic cells, which could be appreciated at different stages of fetal development. The Kupffer cells were also noticed in fetal liver. Conclusion: The present study will be helpful in understanding the normal histology of fetal liver and add to the existing knowledge regarding development of fetal liver.

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Abstract. The present study was planned to investigate the effect of different levels of chlorine dioxide on the performance, gut microbiota and intestinal histology of quails. For this purpose, 300 day-old healthy quail chicks were randomly placed in 20 specially designed pens (15 birds/pen) with water troughs containing chlorine dioxide at the level of 0.00 (DW-0.00), 0.3 (DW-0.3), 0.4 (DW-0.4) and 0.5 ppm (DW-0.5) in replicated fashion (5 replicate/treatment) for 28 days. Weight gain, feed conversion ratio and dressing percentage increased significantly (P<0.05) in DW-0.5 group. Similarly, liver, gizzard and heart weight increased significantly in treated groups linearly with increasing levels of treatment at day 21 and 28. The results showed that population of Salmonella and E. coli decreased linearly at day 21 and 28 of age. Villus height and goblet cells at day 21 and 28 were significantly higher in DW-0.5 group. The results of the present study suggested that the treatment of chlorine dioxide linearly increased the performance and gut morphology and decreased microbial population in quails.

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The present study investigates if the total replacement of dietary fishmeal (FM) with poultry by-product meal (PBM), supplemented with methionine influences the muscle fatty acids composition, normal gut morphology, histological traits of the liver, muscle, and gill, liver enzymes, immune and antioxidant response, and stress-related gene in juvenile barramundi, Lates calcarifer in relation to growth and feed utilization. Barramundi (3.58±0.01g) were randomly distributed into six 300 L seawater recirculating tanks (25 fish/tank) and fed two formulated isonitrogenous and isolipidic diets for 6 weeks. The control diet had FM as the sole animal protein source, whereas other test diet had only PBM as an animal protein source. Dietary PBM affected the fish performance and feed utilization. Regarding muscle fatty acid profile, total saturated fatty acids and monounsaturated fatty acids elevated while total PUFA particularly n-3 LC-PUFA and EPA decreased in PBM fed fish than control diet fed fish. Liver, muscle, gill, and intestinal histology showed no obvious alteration in control diet fed fish, however, more lipid droplets and hepatic vacuolization in the liver, necrotic myotome in muscle, hyperplasia in secondary lamellae in gill and short and broken folds in the intestine were observed in PBM fed fish. Similar to light microscopy observation of intestinal morphology, the transmission electron microscopy (TEM) analysis revealed shorter and smaller microvilli in fish fed PBM. Histopathological alterations in the liver of PBM fed fish were further associated with the elevated levels of aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH) and the significant upregulation of stress-related genes, HSP70 and HSP90. Also, a negative influence on lysozyme activity, and antioxidant enzymatic activities were recorded in fish fed PBM. Overall, it can be concluded that a total substitution of FM protein by methionine supplemented PBM negatively influenced the growth performance, liver health, histological traits of different organs, immune and antioxidant response, and expression of stress-related genes in juvenile barramundi.

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Introduction The objective of the present study was to investigate the in vitro inhibitory effect of the Potentilla fulgens extract on amylase, α- and β-glucosidase, and lipase, as well as its effect on the ultrastructure of the liver, of the kidneys, and of the eye tissues in alloxan-induced diabetic mice. The present study was designed to get further insight regarding the action of P. fulgens from what has been previously known and reported about this plant. Materials and Methods Roots of P. fulgens were extracted with 10 volumes of aqueous-methanol solution (1:4), and the prepared extract was used for in vitro inhibitory activity on amylase, α-glucosidase, β-glucosidase, and lipase. Afterwards, the plant extract was intraperitoneally administered for alternated days (250 mg/kg body weight) to diabetic mice for 4 weeks, and an ultrastructural examination of the liver, the kidneys and the eye tissues was performed using a transmission electron microscope (JEM-100 CX II, Jeol Ltd., Tokyo Japan). Results The P. fulgens extract showed inhibitory activity against all the four enzymes (amylase, α- and β-glucosidase, and lipase), with the highest percentage of inhibition (94.57% ± 0.16 at 1 mg/mL) being observed against α-glucosidase when compared with the standard. The ultrastructural studies revealed a distortion in the structure of the nuclei and of the mitochondria in the kidneys and liver tissues of diabetic mice. Distortion of cell shape and disturbed orientation was observed in the eye lens of diabetic mice. The P. fulgens extract reversed/protected/reduced the ultrastructural alteration observed in the tissues (liver, kidney, and eye lens) of diabetic mice. Conclusion The inhibitory effect of the P. fulgens extract against the aforementioned enzymes and its protective effect on the tissues of diabetic mice against alloxan-induced diabetes add further insight into the antidiabetic properties of this plant.

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Objective: This study was undertaken to evaluate the potential role of Mannan oligosaccharides (MOS) on growth performance, liver and intestine tissue morphology, and gut microbiota of Nile tilapia (Oreochromis niloticus). Design: Controlled study. Animals: Three groups of Nile tilapia. Procedures: Three diets were formulated to contain 0 % of MOS 500 (MOS) as control (control), 0.05% of MOS plus adding MOS into water as well at level of 12.5 mg/l (MOS (feed +water), and the third group MOS was added only to water (MOS water) at the same level, and fed to Nile tilapia for 6 weeks. Results: Simultaneous water and feed additives with (MOS feed+water) at (0.05% of feed + 12.5 mg/l water) caused a significant increase in growth parameters (initial and final weight, weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR) and condition factor (k), when compared to both MOS added only in tank water (MOS water) and control groups. In addition, MOS in both treatment types increased survival rates significantly compared to the control. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) analysis showed that a single species or species that contained chemically equivalent DNA dominated the intestinal tract of all Nile tilapia regardless of dietary treatment. While MOS had no adverse effect of intestinal histology, an increase in the villi length was recorded. Conclusion and clinical relevance: Collectively, our results indicate that MOS added to feed and water could be used to enhance the growth performances, increase the survivability and exert beneficial effects on the gut microbiota of Nile tilapia.

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The paper deals with studying the pathom*orphology of cat pancreas under chronic pancreatitis. This paper is a component of a research mix of the Department of Anatomy and Histology, it goes under the title “The development, morphology and histo-chemistry of animal organs in health and in disease”, (state registration number № 0120U100796). A pancreas is an azygos parenchymatous organ which refers to the endoexocrine glands, includes exocrine and endocrine pancreas, is involved in the processes of digestion and regulation of carbohydrate metabolism, protein metabolism and lipid exchange in tissues. Pancreatic juice, which is rich in enzymes (trypsin, lipase, amylase), is produced in an exocrine pancreas, and hormones (insulin, somatostatin, glucagon (vasoactive intestinal polypeptide), pancreatic polypeptide) are produced in endocrine pancreas. This galand is involved in the process of digestion while producing digestive enzymes, which get into the duodenum and hydrolyze practically all parts of feeds which enter the body. It is located in an abdominal cavity, anatomically connected with a stomach, liver and duedenum. It has been found that pathom*orphological changes in pancreas under chronic pancreatit* manifest themselves depending on the disease stage and are revealed by insignificant progress of the pathological process. Herewith, morphological parameters of pancreas width and length in cats under chronical pancreatitis did not significantly change, but these indices tended to decrease. Its absolute weight in cats under chronical pancreatitis, as compared with clinically healthy cats, did not change and equalled 9.12 ± 2.03 g. But pancreas relative weight in sick cats increased by 1.4 (Р ≤ 0.01) and equalled 0.51 ± 0.08 %, as compared with control 0.38 ± 0.06 %. Under histological analysis of pancreas histology specimen stained with hematoxylin Corazzi and eosin, some distortion in a microscopic structure of a pancreas was observed, it manifested itself in thickening of interparticle tissue-connective layers which spread like desmogenous bands. Some destructive changes in acini in exocrinal pancreas, which manifested themselves in losing their characteristic form, were noticed. The cytoplasm of such acinous cells was in a state of plasmorrhexis, the pycnosis was observed.

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329 Background: CD30 is invariably expressed by untreated EC thus lending support to a rationale for a targeted approach. However, preliminary reports claimed an effect of chemotherapy (CT)-induced downmodulation of CD30 that might affect the potential use in relapsing disease. We aimed at evaluating the persistence of CD30-positivity in post-CT residuals. Methods: We retrieved paraffin-embedded post-CT samples yielding non-teratoma viable cells after ≥ 1 CDDP-based CT from the institutional surgical series. EC component was required and assessed by morphology and Oct-3/4 staining. The entire set was re-stained for CD30 and assessed by 2 pathologists blinded to study purpose. CD30-positivity was defined as a >80% membranous staining with a diffuse moderate-to-strong intensity. Clinical data included site of tumor primary, histology, CT regimen, and type of surgery. Results: From 12/1990 to 09/2012, a total of 246 cases with pure EC or mixed GCT residuals were treated. 49 (EC: 16; mixed GCT: 33) had both complete data and suitable tissue for study purposes. 40 pts had retroperitoneal or mediastinal nodes, 12 pts had lung metastases, 4 had either liver, bone or brain mets. 35/49 cases (70%) preserved CD30 positivity. 20/35 (57%) pts had residual disease after 1stline CT, 15/35 (43%) after multiple CTs (median 3.5, range 2-5). 2 pts (6%) had undergone high-dose CT and 4/35 (11%) were late relapses. 32/35 pts (91%) had gonadal primary, 1 had a retroperitoneal and another a mediastinal primary. The median survival of 35 CD30-positive patients was 16 mos (IQR 3.3-22.9) while it was 48.5 mos (IQR 21.7-72.6) for the 14 CD30-negative patients (p=0.0297 at Gehan-Breslow-Wilcoxon test). 5-year overall survival of CD30-positive and negative patients was 35.7% (95% CI: 18.2-53.6) and 49.7% (95% CI: 19.7-74.0), respectively. Conclusions: Our results on selected relapsing pts suggest that EC retained CD30 even in the far salvage setting, thus confirming to be a reliable target for treatment. A trend towards a poorer prognosis and shorter survival characterized CD30+ cases. A phase 2 study with the antibody-drug conjugate Brentuximab vedotin is currently in plan for GCT.

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BACKGROUNDThe long term outcome of drug related liver disease is unknown.AIMSTo study the natural history of histologically proved drug induced hepatotoxicity.METHODS110 patients with liver biopsies coded either as drug induced liver disease or hepatitis/cholestasis of unknown aetiology were identified from hospital records 1978–1996. Review of case notes and histology identified 44 patients with definite drug induced hepatotoxicity. Forty surviving patients were invited to attend a follow up clinic. History, examination, full liver screen, and isotope and ultrasound liver scans were repeated in all patients. Repeat liver biopsies were offered to patients with abnormal liver tests.RESULTSPresentation at index biopsy was jaundice in 24 patients, abnormal liver tests in 17, and hepatic failure in three. Antibiotics (n=13) and non-steroidal anti-inflammatory drugs (n=11) were the most common drugs implicated. Initial histology showed acute hepatitis in six, chronic hepatitis in 20, and cholestasis in 18. At 1–19 years (median 5 years) follow up, 13/33 (39%) patients had persistent significant abnormalities in their liver blood tests and/or scans. Three of the five repeat liver biopsies performed showed significant abnormalities. Factors predicting persistence or development of chronic liver disease were fibrosis and continued exposure to the drug.CONCLUSIONSDrugs should be considered in the differential diagnosis of abnormal liver function and/or histology, as failure to withdraw the offending drug is associated with a high risk of persistent liver damage.

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Background—A novel flavivirus has been described recently and designated hepatitis G virus (HGV). The virus is transmitted by the parenteral route but it is uncertain whether it is associated with chronic liver disease because liver biopsy is difficult to justify in this group.Aims—To examine histological features of liver biopsy in patients infected with hepatitis C virus (HCV) according to the presence or absence of HCV and HGV RNA.Methods—One hundred and thirty one consecutive HCV carriers undergoing staging liver biopsy were studied retrospectively. In each, HCV RNA and HGV RNA were detected by reverse transcription polymerase chain reaction on serum samples collected at the time of biopsy. The presence of each RNA was correlated with histological features blind to the RNA results; individual histological features of inflammation or fibrosis were scored separately.Results—Nineteen patients were positive for both HGV and HCV RNA in serum, 91 were positive for HCV RNA alone, two were positive for HGV RNA alone, and 19 were negative for both RNA species. Neither age nor sex differed between the groups; a greater proportion of intravenous drug users were HGV RNA positive, but this was not statistically significant. There was no effect of HGV coinfection on the stage of fibrosis or any other histological parameter except steatosis; patients with HCV and HGV RNA had a higher mean score for fat than those patients with HCV RNA alone (p<0.05).Conclusions—HGV coinfection has no important effects on histological features in chronic HCV carriers. It is unlikely that HGV infection causes chronic liver disease.

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The present study was conducted to evaluate the effects of probiotic supplementation on theintestinal morphology of Thai pangasius (Pangasianodonhypophthalmus). The experiment was conducted in15 earthen ponds of 0.75 dec. each. Each pond contained 600 fingerlings/decimal(5.69±1.23 g) and thefeeding trial was performed for 90 days. Commercially available gut nutrient additive, Mutagen and pHFIXER (water additive probiotic) were used in the experiment. Five treatments were conducted each havingthree replicates.Treatment 1 (T1) was provided with gut probiotics, treatment 2 (T2) with water probiotics,Treatment 3 (T3) with50% gut probiotic and 50% water probiotic; Treatment 4 (T4) with prebiotic (yeast andrice starch) and 50% gut and 50% water probiotic.Treatment 5 (T5) was designed as control. Fishes were fedtwice a day at the rate of 10-5% of body weight. Samplings were carried out fortnightly where gut sampleswere collected for histology and water quality parameters were recorded. All water quality parameters werefound to be favorable for fish culture. Histological analysis of the gastrointestinal tract revealed that foldlength, width and enterocyte height were significantly larger (p<0.05) in fish of T3 and T4compared to thoseof T1, T2, T5. Thus, probiotic supplementation can improve the intestinal morphology of Thai pangasiuswhich may increase the nutrient absorption in fish and thereby increase the digestion capacity.

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Recently, the second mammalian chitinase, designated acidic mammalian chitinase (AMCase), has been identified in human, mouse, and cow. In contrast to the earlier identified macrophage-derived chitinase (chitotriosidase), this chitinase is richly expressed in the gastrointestinal (GI) tract, suggesting its role in digestion of chitin-containing foods as well as defense against chitin-coated microorganisms and parasites. This in situ hybridization study first revealed cellular localization of the gut-type chitinase in the mouse and chicken. In adult mice, the parotid gland, von Ebner's gland, and gastric chief cells, all of which are exocrine cells of the serous type, expressed the gut chitinase mRNA. In the chicken, oxyntico-peptic cells in glandular stomach (proventriculus) and hepatocytes expressed the chitinase mRNA. Because cattle produce the gut chitinase (chitin-binding protein b04) only in the liver, the gut chitinases in mammals and birds have three major sources of production, i.e., the salivary gland, stomach, and liver. During ontogenetic development, the expression level in the parotid gland and stomach of mice increased to the adult level before weaning, whereas in the stomach of chickens intense signals were detectable in embryos from incubation day 7.

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SUMMARYStudies have been carried out to establish the relative importance of oral and trans-tegumental uptake of triclabendazole by the liver fluke,Fasciola hepatica. Experiments were designed to block either oral uptake of drug, by use of ligatures, or trans-tegumental diffusion, by allowing the drug to bind to bovine serum albumin (BSA) in the medium. Changes to the surface morphology of the tegument and gut were assessed by scanning electron microscopy. Flukes were incubatedin vitrofor 24 h in TCBZ.SO at a concentration of 15 μg/ml. Tegumental disruption in ligatured and non-ligatured flukes was similar, suggesting that closing the oral route did not affect drug uptake. The gut remained unaffected by drug treatment. When BSA (30 mg/ml) was present in the medium, there was a marked decline in the level of tegumental disruption. Again, the gut retained a normal morphology. Non-ligatured flukes were also incubated for 24 hin vitroin TCBZ.SO (15 μg/ml) in the presence of red blood cells. Oral ingestion of blood was demonstrated, although the gut surface retained a normal morphology. In contrast, the tegumental surface was severely affected by the drug. The findings support previous pharmacological studies which suggest that trans-tegumental uptake of triclabendazole predominates in the liver fluke.

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The mechanisms whereby exogenous growth hormone modulates intestinal structure and function in fish were investigated. Goldfish (Carassius auratus) were fed commercial flake diet sprayed with recombinant carp growth hormone (cGH) daily for 1 month. Control animals received food sprayed with the vehicle. After 1 month of daily feedings, body mass and length were determined, and animals were sacrificed to study intestinal characteristics. Sections of foregut were removed after determination of total gut length for measurement of leucine uptake, histology, and epithelial ultrastructure. Oral administration of cGH for 1 month resulted in a 40% increase in body mass and an 8% increase in body length above controls. Gut length was 43% greater and the gut length to body length ratio was 32% greater as a result of the cGH treatment. Feeding with cGH also resulted in a significant increase in leucine uptake and increased gut mucosal thickness. Analysis of transmission electron micrographs revealed significant increases in the microvillous height and density and epithelial surface area. The findings indicate that growth hormone added to feed may increase growth in fish, in part by significantly increasing gut length, mucosal thickness, and epithelial brush border surface area, leading to enhanced epithelial absorption.Key words: growth hormone, goldfish, intestine, morphology, leucine.

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A patient presented with pruritus and recent elevation of aminotransferases. The case fulfilled most of the criteria for the diagnosis of autoimmune hepatitis and achieved clinical and complete biochemical response to steroid therapy. However, the liver biopsy specimen revealed an unusual histological pattern consisting of severe centrilobular necrosis demarcated by a thin rim of hepatitic reaction. In contrast, the portal tracts appeared almost normal. This histological appearance has not been associated with autoimmune hepatitis. This presentation and the histology may represent an early pattern of autoimmune injury to the liver.

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Serum vitamin D levels negatively correlate with obesity and associated disorders such as non-alcoholic steatohepatitis (NASH). However, the mechanisms linking low vitamin D (VD) status to disease progression are not completely understood. In this study, we analyzed the effect of VD treatment on NASH in mice. C57BL6/J mice were fed a high-fat/high-sugar diet (HFSD) containing low amounts of VD for 16 weeks to induce obesity, NASH and liver fibrosis. The effects of preventive and interventional VD treatment were studied on the level of liver histology and hepatic/intestinal gene expression. Interestingly, preventive and to a lesser extent also interventional VD treatment resulted in improvements of liver histology. This included a significant decrease of steatosis, a trend towards lower non-alcoholic fatty liver disease (NAFLD) activity score and a slight non-significant decrease of fibrosis in the preventive treatment group. In line with these changes, preventive VD treatment reduced the hepatic expression of lipogenic, inflammatory and pro-fibrotic genes. Notably, these beneficial effects occurred in conjunction with a reduction of intestinal inflammation. Together, our observations suggest that timely initiation of VD supplementation (preventive vs. interventional) is a critical determinant of treatment outcome in NASH. In the applied animal model, the improvements of liver histology occurred in conjunction with reduced inflammation in the gut, suggesting a potential relevance of vitamin D as a therapeutic agent acting on the gut–liver axis.

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This study was undertaken to identify Fasciolagiganticai on the basis of its morphology and histology to be the common cause of fasciolosis in infected buffaloes. Material & Method: Adult liver flukes were recovered from the liver of naturally infected buffaloes slaughtered in various abattoirs in Gujarat. Some adult flukes were flattened, put between two slides , pressed and stained in Borax carmine, and some flukes were sectioned in the median sagittal plane and histological slides of the flukes were prepared for detailed morphological and histological studies. Result: Microscopic pictures of the parasite used in identification defines the similarity in the morphology and histology of the F. gigantica on the basis of morphology of flukes; anterior sucker, posterior sucker (acetabulum), pharynx, uterus, ovary and type of epithelium. Conclusion: It can be concluded that the most common species found in buffaloes infected with Fasciolagigantica on the basis of its histo-morphological appearance in Gujarat.

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As the most common grain contaminant worldwide, deoxynivalenol is of high importance despite its low toxicity compared to other trichothecene mycotoxins. Data on the effects of deoxynivalenol in rabbits are scarce. Thus, the aim of this study was to investigate the effects of dietary deoxynivalenol fed at a high level (10 mg/kg of feed) on the productive performance, blood indices, immunological variables, histopathological changes, and genotoxicity in rabbits. Forty-eight Pannon White rabbits were exposed to contaminated diets for three weeks. Despite its high concentration, deoxynivalenol did not affect the feed intake, body weight, and body weight gain. Liver and kidney function was not affected, as shown by the clinical chemistry indices. Conversely, in two rabbits the toxin caused mild fibrosis of the liver, without degenerative changes of the hepatocytes. No genotoxicity could be observed either. Gut cytokines and the phagocytic activity of the macrophages did not differ significantly. The percentage of neutrophils was significantly lower, whereas that of eosinophils was significantly higher in the toxin-fed group. Deoxynivalenol did not cause significant changes in gut and villus morphology. In 4 out of the 6 deoxynivalenol-treated animals, the ratio of lymphoblast proliferation and simultaneous apoptosis shifted towards apoptosis in the gut-associated lymphoid tissue. In the central part of the lymphoid follicles of the spleen, lymphocyte depletion and follicular atrophy could be detected. It can be concluded that rabbits are less sensitive to deoxynivalenol, but the findings confirm that this Fusarium toxin is capable of modulating the immune response.

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Fenestrations are pores within the liver sinusoidal endothelial cells (LSECs) that line the sinusoids of the highly vascularized liver. Fenestrations facilitate the transfer of substrates between blood and hepatocytes. With pseudocapillarization of the hepatic sinusoid in old age, there is a loss of fenestrations. LSECs are uniquely exposed to gut-derived dietary and microbial substrates delivered by the portal circulation to the liver. Here we studied the effect of 25 diets varying in content of macronutrients and energy on LSEC fenestrations using the Geometric Framework method in a large cohort of mice aged 15 mo. Macronutrient distribution rather than total food or energy intake was associated with changes in fenestrations. Porosity and frequency were inversely associated with dietary fat intake, while fenestration diameter was inversely associated with protein or carbohydrate intake. Fenestrations were also linked to diet-induced changes in gut microbiome, with increased fenestrations associated with higher abundance of Firmicutes and reduced abundance of Bacteroidetes. Diet-induced changes in levels of several fatty acids (C16:0, C19:0, and C20:4) were also significantly inversely associated with fenestrations, suggesting a link between dietary fat and modulation of lipid rafts in the LSECs. Diet influences fenestrations and these data reflect both the key role of the LSECs in clearing gut-derived molecules from the vascular circulation and the impact these molecules have on LSEC morphology.

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Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in western countries, with a continuously rising incidence. Gut-liver communication and microbiota composition have been identified as critical drivers of the NAFLD progression. Hence, it has been shown that microbiota depletion can ameliorate high-fat diet or western-diet induced experimental Non-alcoholic steatohepatitis (NASH). However, its functional implications in the methionine-choline dietary model, remain incompletely understood. Here, we investigated the physiological relevance of gut microbiota in methionine-choline deficient (MCD) diet induced NASH. Experimental liver disease was induced by 8 weeks of MCD feeding in wild-type (WT) mice, either with or without commensal microbiota depletion, by continuous broad-spectrum antibiotic (AB) treatment. MCD diet induced steatohepatitis was accompanied by a reduced gut microbiota diversity, indicating intestinal dysbiosis. MCD treatment prompted macroscopic shortening of the intestine, as well as intestinal villi in histology. However, gut microbiota composition of MCD-treated mice, neither resembled human NASH, nor did it augment the intestinal barrier integrity or intestinal inflammation. In the MCD model, AB treatment resulted in increased steatohepatitis activity, compared to microbiota proficient control mice. This phenotype was driven by pronounced neutrophil infiltration, while AB treatment only slightly increased monocyte-derived macrophages (MoMF) abundance. Our data demonstrated the differential role of gut microbiota, during steatohepatitis development. In the context of MCD induced steatohepatitis, commensal microbiota was found to be hepatoprotective.

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Thioacetamide (TAA) is widely used to study liver toxicity accompanied by oxidative stress, inflammation, cell necrosis, fibrosis, cholestasis, and hepatocellular carcinoma. As an efficient free radical’s scavenger, C60 fullerene is considered a potential liver-protective agent in chemically-induced liver injury. In the present work, we examined the hepatoprotective effects of two C60 doses dissolved in virgin olive oil against TAA-induced hepatotoxicity in rats. We showed that TAA-induced increase in liver oxidative stress, judged by the changes in the activities of SOD, CAT, GPx, GR, GST, the content of GSH and 4-HNE, and expression of HO-1, MnSOD, and CuZnSOD, was more effectively ameliorated with a lower C60 dose. Improvement in liver antioxidative status caused by C60 was accompanied by a decrease in liver HMGB1 expression and an increase in nuclear Nrf2/NF-κB p65 ratio, suggesting a reduction in inflammation, necrosis and fibrosis. These results were in accordance with liver histology analysis, liver comet assay, and changes in serum levels of ALT, AST, and AP. The changes observed in gut microbiome support detrimental effects of TAA and hepatoprotective effects of low C60 dose. Less protective effects of a higher C60 dose could be a consequence of its enhanced aggregation and related pro-oxidant role.

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The gut microbiota plays a role in nonalcoholic fatty liver disease (NAFLD), but data about gut dysbiosis in Asians with NAFLD remains scarce. We analyzed the differences in fecal microbiota between adults with and without NAFLD. This cross-sectional study examined adults with histology-proven NAFLD (25 nonalcoholic fatty liver (NAFL) patients, 25 nonalcoholic steatohepatitis (NASH) patients, and 25 living liver donors (healthy controls)). The taxonomic composition of the gut microbiota was determined by 16S ribosomal RNA gene sequencing of stool samples. The NAFL and NASH groups showed lower total bacterial diversity and richness than the controls. NAFLD patients had higher levels of the phylum Bacteroidetes and lower levels of Firmicutes than controls. The genus Ruminococcaceae UCG-010, family Ruminococcaceae, order Clostridiales, and class Clostridia were less abundant in patients with NAFL or NASH than healthy individuals. The lipopolysaccharide biosynthesis pathway was differentially enriched in the NASH group. This study examined the largest number of Asian patients with biopsy-proven NAFL and NASH in terms of dysbiosis of the gut microbiota in NAFLD patients. NAFLD patients had higher levels of Bacteroidetes and lower levels of Firmicutes. These results are different from research from western countries and could provide different targets for therapies by region.

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A compared study of the morphology of the alimentary tract and liver of seven selected species corresponding to the main clades of the Characidae family is presented herein. Three new set of characters corresponding to 1) alimentary tract gross anatomy, 2) alimentary tract histology and 3) liver gross anatomy are evaluated as potential sources of data for future phylogenetic studies of the Characidae. Not considerable interspecific variation was observed at the histological level and therefore this source is not considered to be phylogenetically informative at the taxonomic level analyzed. In contrast, liver and alimentary tract gross anatomy presented important interspecific variation while a relatively hom*ogenous intraspecific morphology was observed. Those characters are optimized in tree-topologies from previous phylogenetic analyses and their evolution and potential relationship with ecological traits are discussed.

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The purpose of this study was to determine the effects of single and combined administrations of deoxynivalenol (DON) and zearalenone (ZEN) on the histology and ultrastructure of pig liver. The study was performed on immature gilts, which were divided into four equal groups. Animals in the experimental groups received DON at a dose of 12 μg/kg body weight (BW) per day, ZEN at 40 μg/kg BW per day, or a mixture of DON (12 μg/kg BW per day) and ZEN (40 μg/kg BW). The control group received vehicle. The animals were killed after 1, 3, and 6 weeks of experiment. Treatment with mycotoxins resulted in several changes in liver histology and ultrastructure, including: (1) an increase in the thickness of the perilobular connective tissue and its penetration to the lobules in gilts receiving DON and DON + ZEN; (2) an increase in the total microscopic liver score (histology activity index (HAI)) in pigs receiving DON and DON + ZEN; (3) dilatation of hepatic sinusoids in pigs receiving ZEN, DON and DON + ZEN; (4) temporary changes in glycogen content in all experimental groups; (5) an increase in iron accumulation in the hepatocytes of gilts treated with ZEN and DON + ZEN; (6) changes in endoplasmic reticulum organization in the hepatocytes of pigs receiving toxins; (7) changes in morphology of Browicz–Kupffer cells after treatment with ZEN, DON, and DON + ZEN. The results show that low doses of mycotoxins used in the present study, even when applied for a short period, affected liver morphology.

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Background and aims—The liver is frequently involved in amyloidosis but the significance of hepatic amyloid has not been systematically studied. We have previously developed scintigraphy with 123I serum amyloid P component (123I-SAP) to identify and monitor amyloid deposits quantitatively in vivo and we report here our findings in hepatic amyloidosis.Methods—Between 1988 and 1995, 805 patients with clinically suspected or biopsy proven systemic amyloidosis were evaluated. One hundred and thirty eight patients had AA amyloidosis, 180 had AL amyloidosis, 99 had hereditary amyloid syndromes, and 67 had dialysis related (β2 microglobulin) amyloid. One hundred and ninety two patients with amyloidosis were followed for six months to eight years.Results—Hepatic amyloid was found in 98/180 (54%) AL and 25/138 (18%) AA patients but in only 1/53 patients with familial transthyretin amyloid polyneuropathy and in none with dialysis related amyloidosis. There was complete concordance between hepatic SAP scintigraphy and the presence or absence of parenchymal amyloid deposits on liver histology. Amyloidosis was never confined to the liver. Mortality was rarely due to hepatic failure, although hepatic involvement with AA amyloid carried a poor prognosis. Successful therapy to reduce the supply of amyloid fibril protein precursors was followed by substantial regression of all types of amyloid.Conclusions—SAP scintigraphy is a specific and sensitive method for detecting and monitoring hepatic amyloid. Liver involvement is always associated with major amyloid in other organ systems and carries a poor prognosis in AA type. Appropriate therapy may substantially improve prognosis in many patients.

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The development of effective strategies for local administration of chemotherapeutic drugs, thus minimizing the adverse side effects to patients, is one of the key challenges in biomedicine and cancer research. This work reports the formulation and characterization of PluronicF127 microemulsions to enhance the bioavailability of Cisplatin (Cis). The size of Cis microemulsion was about 12.0 nm, as assessed by dynamic light scattering analysis. In vitro cytotoxic activity of free Cis and F127/Cis microemulsions were studied on malignant (C152 and MCF7) and normal (HUVEC) cells via tetrazolium (MTT) colorimetric assay. Cell morphology was also monitored. In vitro assessments revealed thatF127/Cis microemulsions induced cytotoxicity/morphological changes to a lesser extent than free Cis. Regarding in vivo experiments, F127/Cis microemulsions were injected intraperitoneally at 7 and 14 mg/kg doses into adult male Wistar rats to assess histologic and biochemical changes. In this case, the bulk Cis group caused severe histopathological changes and significant increases in serum liver enzymes and serum kidney function markers. The group treated with the 14 mg/kg dose of F127/Cis microemulsions also showed severe fatty changes and significant increases in serum liver enzymes, blood urea nitrogen, and creatinine levels. The group treated with the low dose of nano-Cis showed a significant increase in serum liver enzymes levels accompanied by mild fatty changes of the liver. Theoretical surveys were performed to get an understanding of the interplay between F127 and Cis. Results reveal that hydrogen bonding (HB) interactions with F127have an influence on the molecular properties of Cis and may playa role in the lower toxicity of F127/Cis in comparison to free Cis.

Abstract:

When the aquaculture water environment deteriorates or the temperature rises, shrimp are susceptible to viral or bacterial infections, causing a large number of deaths. This study comprehensively evaluated the effects of the oral administration of a chitosan–gentamicin conjugate (CS-GT) after Litopenaeus vannamei were infected with Vibrio parahaemolyticus, through nonspecific immunity parameter detection, intestinal morphology observation, and the assessment of microbial flora diversification by 16S rRNA gene sequencing. The results showed that the oral administration of CS-GT significantly increased total hemocyte counts and reduced hemocyte apoptosis in shrimp (p < 0.05). The parameters (including superoxide dismutase, glutathione peroxidase, glutathione, lysozyme, acid phosphatase, alkaline phosphatase, and phenoloxidase) were significantly increased (p < 0.05). The integrity of the intestinal epithelial cells and basem*nt membrane were enhanced, which correspondingly alleviated intestinal injury. In terms of the microbiome, the abundances of Vibrio (Gram-negative bacteria and food-borne pathogens) in the water and gut were significantly reduced. The canonical correspondence analysis (CCA) showed that the abundances of Vibrio both in the water and gut were negatively correlated with CS-GT dosage. In conclusion, the oral administration of CS-GT can improve the immunity of shrimp against pathogenic bacteria and significantly reduce the relative abundances of Vibrio in aquaculture water and the gut of Litopenaeus vannamei.

Abstract:

BackgroundThe pathogenesis of non-alcoholic fatty liver disease (NAFLD) is incompletely understood. Kupffer cells (KCs), phagocytic liver-resident macrophages, provide a protective barrier against egress of endotoxin from the portal to the systemic circulation. It is not known if KC phagocytic function is impaired in NAFLD. Super-paramagnetic iron oxide (SPIO) magnetic resonance imaging is a comparative technology dependent on KC phagocytic function.ObjectiveTo evaluate KC uptake function, in patients and experimental animals with NAFLD, using SPIO.MethodsAbdominal CT and histological examination of liver biopsy specimens were used to estimate the degree of steatosis in patients with NAFLD and controls with chronic hepatitis C. SPIO-MRI was then performed in all patients. Normal rats fed a methionine-choline-deficient diet to induce non-alcoholic steatohepatitis (NASH), the more severe stage of NAFLD, and obese, insulin resistant, Zucker fa/fa rats with steatohepatitis, were also studied with SPIO-MRI and analysed for hepatic uptake of fluorescent microbeads. Immunohistochemical analysis evaluated the numbers of KCs in patients and rat livers.ResultsRelative signal enhancement (RSE), inversely proportional to KC function, was higher in patients with NAFLD than in controls and with the degree of steatosis on CT. RSE also positively correlated with the degree of steatosis on histology and was similarly higher in rats with induced severe NAFLD (NASH). On immunohistochemistry, defective phagocytic function was the result of reduced phagocytic uptake and not due to reduced KC numbers in rats or patients with NAFLD.ConclusionsKC uptake function is significantly impaired in patients with NAFLD and experimental animals with NASH, worsens with the degree of steatosis and is not due to a reduction of KC numbers.

Abstract:

The trial investigates if a highly defatted Hermetia illucens larva meal (H) at two dietary inclusion levels and a vegetable protein based diet (VEG) influences the normal gut and liver histology and the oxidative stress biomarkers in liver and kidney of Siberian sturgeon juveniles. Fish were fed four diets: one control diet (H0) containing 70% of fishmeal (FM), two diets including 18.5% (H185) and 37.5% (H375) of highly defatted H in substitution for 25% and 50% of FM, and one vegetable protein based diet (VEG). At the end of a growth trial, 12 fish per treatment were sacrificed by over-anaesthesia to collect 12 liver and 5 distal intestine samples for histological analyses, as well as 12 liver and kidney samples for biochemical analyses. The H and VEG diets did not significantly affect the histology of liver and distal intestine, but alterations of the oxidative stress biomarkers were detected at the highest inclusion level of H (37.5%). In order to avoid unfavorable effects on the fish health, an inclusion level up to 18.5% of H is recommended for Siberian sturgeon juveniles.

Abstract:

The liver is an organ notable for its sensitivity to a great variety of environmental factors. It is composed of a parenchyma divided into irregular lobules by the exocrine pancreas or hepatopancreas; the pancreatic cells are arranged around a branch of the portal vein. The hepatocytes are radially arranged in cords around a central sinusoid. The liver histomorphology and the organization of exocrine pancreatic tissue of O. jenynsii (Günther, 1864) is similar to the acinar morphology of many teleosts, including freshwater and marine species. The aim of the present work was to carry out the histological analysis of the liver of Oligosarcus jenynsii, one of the most common species inhabiting Los Padres Lake (Buenos Aires Province, Argentina).

Abstract:

Background. Accumulative evidence showed that gut microbiota was important in regulating the development of nonalcoholic fatty liver disease (NAFLD). Hugan Qingzhi tablet (HQT), a lipid-lowering and anti-inflammatory medicinal formula, has been used to prevent and treat NAFLD. However, its mechanism of action is unknown. The aim of this study was to confirm whether HQT reversed the gut microbiota dysbiosis in NAFLD rats. Methods. We established an NAFLD model of rats fed with a high-fat diet (HFD), which was given different interventions, and measured the level of liver biochemical indices and inflammatory factors. Liver tissues were stained with hematoxylin-eosin and oil red O. Changes in the gut microbiota composition were analyzed using 16S rRNA sequencing. Results. The hepatic histology and biochemical data displayed that HQT exhibited protective effects on HFD-induced rats. Moreover, HQT also reduced the abundance of the Firmicutes/Bacteroidetes ratio in HFD-fed rats and modified the gut microbial species at the genus level, increasing the abundances of gut microbiota which were reported to have an effect on relieving NAFLD, such as Ruminococcaceae, Bacteroidales_S24-7_group, Bifidobacteria, Alistipes, and Anaeroplasma, and significantly inhibiting the relative abundance of Enterobacteriaceae, Streptococcus, Holdemanella, Allobaculum, and Blautia, which were reported to be potentially related to NAFLD. Spearman’s correlation analysis found that [Ruminococcus]_gauvreauii_group, Lachnoclostridium, Blautia, Allobaculum, and Holdemanella exhibited significant (p<0.001) positive correlations with triglyceride, cholesterol, low-density lipoprotein cholesterol, interleukin-6, interleukin-1β, tumor necrosis factor-α, and body weight and negative correlations with high-density lipoprotein cholesterol (p<0.001). The norank_f__Bacteroidales_S24-7_group and Alistipes showed an opposite trend. Moreover, the HQT could promote flavonoid biosynthesis compared with the HFD group. Conclusion. In summary, the HQT has potential applications in the prevention and treatment of NAFLD, which may be closely related to its modulatory effect on the gut microbiota.

Abstract:

This study was conducted to compare the effects of using direct fed microbial (RE3) and antibiotics on the gut morphology. Two hundred and forty unsexed, one-day old Marshal strain broiler chicks were used for the experiment. The birds were randomly allotted into three dietary treatments with each treatment replicated four times at 20 birds per replicate making a total of 80 birds per treatment. The experimental design was completely randomized design. The villous height (9.303E2), the crypt depth (1.5053E2) and the muscular thickness (2.2311E2) of the RE3 treated birds were higher (P<0.05) when compared with birds fed with control diet (T1) 8.619E2, 1.3790E2 and 1.9645E2 and the control + antibiotics (T3) 7.0677E2, 1.3331E2 and 1.9027E2 respectively. The observations revealed that birds put on treatment 2 (RE3 probiotics) had better presentation and preservation of the intestinal villi, glands and intestinal wall integrity. It was concluded that supplementation of broilers diets with the direct fed microbial (RE3) will lead to maintenance of intestinal health and better utilization of nutrients to enable full expression of genetic potential.

Abstract:

Summary Cholestasis is a condition wherein bile flow is interrupted and lithocholic acid is known to play a key role in causing severe liver injury. In this study, we performed in-depth analysis of the morphological changes in bile canaliculi and the biological role of villin in cholestasis using lithocholic acid-stimulated HepG2 human hepatocarcinoma cells. We confirmed disruption of the bile canaliculi in liver sections from a liver allograft patient with cholestasis. Lithocholic acid caused strong cytotoxicity in HepG2 cells, which was associated with abnormal morphology. Lithocholic acid reduced villin expression, which recovered in the presence of nuclear receptor agonists. Furthermore, villin mRNA expression increased following small interfering RNA (siRNA)-mediated knockdown of the nuclear farnesoid X receptor and pregnane X receptor. Villin knockdown using siRNA caused cell growth arrest in HepG2 cells. The effect of villin-knockdown on whole-genome expression in HepG2 cells was analyzed by DNA microarray. Our data suggest that lithocholic acid caused cell growth arrest by suppressing villin expression via farnesoid X receptor and pregnane X receptor in HepG2 cells.

Abstract:

Angiomyxomas are uncommon myxoid tumours arising most commonly from the pelvis. A 46-year-old woman with a history of polycystic kidney disease presented asymptomatically for surveillance ultrasonography; changes were noted in the size and morphology of her liver cysts. Subsequent radiological assessment displayed features suspicious of malignancy and a right hemihepatectomy was performed with curative intent. Pathological examination of the resected specimen found histology consistent with an angiomyxoma arising primarily from the liver parenchyma. Follow-up review of the patient has been uneventful with annual imaging showing no evidence of recurrence. Angiomyxomas do not characteristically invade other tissues. However, any liver lesion displaying suspicious features of malignancy should be resected in the absence of disseminated disease.

Abstract:

This study was conducted to estimate the effect of adding different levels of turmeric powder and curcumin on broiler performance, carcass traits, and immunity response and jejunum histology. A total of 750 Ross day old chicks obtained from a local hatchery and distributed into two rearing conditions (normal and heat stress condition) with five treatments 3 replicate for each one had (25 chicks). Dietary supplements of turmeric powder and curcumin significantly (P ≤ 0.05) improved performance under normal and heat stress conditions. Moreover, treatments significantly (P ≤ 0.05) increased the total titter antibody against Newcastle and infectious bursal disease compared to control. Concerning histology of jejunum treatments significantly (P≤0.01) improved jejunum histology. Regarding the effect of rearing condition heat stress conditions significantly (P ≤ 0.01) decreased performance compared to normal rearing condition. Small intestine percent from live body weight significantly (P ≤ 0.01) decreased compared to normal rearing conditions. Moreover, heat stress significantly (P≤0.01) affected the total antibody titer against Newcastle disease and infectious bursal disease. Jejunum histology significantly (P ≤ 0.01) affected by rearing conditions.

Abstract:

Valine is an important essential amino acid of laying hens. Dietary supplemented with BCAAs ameliorated gut microbiota, whereas elevated blood levels of BCAAs are positively associated with obesity, insulin resistance, and diabetes in both humans and rodents. General controlled nonrepressed (GCN2) kinase plays a crucial role in regulating intestinal inflammation and hepatic fatty acid homeostasis during amino acids deficiency, while GCN2 deficient results in enhanced intestinal inflammation and developed hepatic steatosis. However, how long-term dietary valine impacts gut health and the development of nonalcoholic fatty liver disease (NAFLD) remains unknown. Hence, in the present study, we elucidated the effects of dietary valine on intestinal barrier function, microbial homeostasis, and the development of NAFLD. A total of 960 healthy 33-weeks-old laying hens were randomly divided into five experimental groups and fed with valine at the following different levels in a feeding trial that lasted 8 weeks: 0.59, 0.64, 0.69, 0.74, and 0.79%, respectively. After 8 weeks of treatment, related tissues and cecal contents were obtained for further analysis. The results showed that diet supplemented with valine ameliorated gut health by improving intestinal villus morphology, enhancing intestinal barrier, decreasing cecum pathogenic bacteria abundances such as Fusobacteriota and Deferribacterota, and inhibiting inflammatory response mediated by GCN2. However, long-term intake of high levels of dietary valine (0.74 and 0.79%) accelerated the development of NAFLD of laying hens by promoting lipogenesis and inhibiting fatty acid oxidation mediated by GCN2-eIF2α-ATF4. Furthermore, NAFLD induced by high levels of dietary valine (0.74 and 0.79%) resulted in strengthening oxidative stress, ER stress, and inflammatory response. Our results revealed that high levels of valine are a key regulator of gut health and the adverse metabolic response to NAFLD and suggested reducing dietary valine as a new approach to preventing NAFLD of laying hens.

Abstract:

Abstract Possible liver damage induced by chemicals or drugs must be detected early during drug development or industrial exposure, although damage is still difficult to predict, especially when immunotoxicity is involved. Liver toxicity may result from cytolysis, steatosis, cholestasis, phospholipidosis, or vascular lesions, most the outcome of a disadvantageous balance between chemicals or metabolites vs protective mechanisms, resulting from chemical dosage, genetic factors, or the immunoallergic status of the patient. Drug metabolism, lipid peroxidation, and thiol oxidation are frequently involved in liver toxicities. Classical guidelines in toxicology propose many methods for liver toxicity assessment: histology; chemical changes in hepatic tissue (lipids, glutathione, enzymes); physiological changes in biosynthesis (proteins, glycoproteins); excretion function (fructose); drug metabolism; and concentrations of related enzymes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyltransferase) in blood. In vitro studies in human or animal hepatocytes or tumor-derived cell lines are useful in detecting hepatocellular lesions by cell viability, glutathione concentration, amount of lactate dehydrogenase released, cellular ATP, morphology (blebs), and drug metabolism.

Abstract:

The relationship between gender and alcohol-induced liver disease is complex; however, endotoxin is most likely involved. Recently, it was reported that estriol activated Kupffer cells by upregulation of the endotoxin receptor CD14. Therefore, the purpose of this work was to study how estriol sensitizes Kupffer cells. Rats were given estriol (20 mg/kg ip), and Kupffer cells were isolated 24 h later. After addition of lipopolysaccharide (LPS), intracellular Ca2+ concentration was measured using a microspectrofluorometer with the fluorescent indicator fura 2, and tumor necrosis factor-α was measured by ELISA. CD14 was evaluated by Western analysis. One-half of the rats given estriol intraperitoneally 24 h before an injection of a sublethal dose of LPS (5 mg/kg) died within 24 h, whereas none of the control rats died. Mortality was prevented totally by sterilization of the gut with antibiotics. A similar pattern was obtained with liver histology and serum transaminases. Translocation of horseradish peroxidase was increased about threefold in gut segments by treatment with estriol. This increase was not altered by treatment with nonabsorbable antibiotics. On the other hand, endotoxin levels were increased to 60–70 pg/ml in plasma of rats treated with estriol. As expected, this increase was prevented (<20 pg/ml) by antibiotics. In isolated Kupffer cells, LPS-induced increases in intracellular Ca2+ concentration, tumor necrosis factor-α production, and CD14 were increased, as previously reported. All these phenomena were blocked by antibiotics. Therefore, it is concluded that estriol treatment in vivo sensitizes Kupffer cells to LPS via mechanisms dependent on increases in CD14. This is most likely due to elevated portal blood endotoxin caused by increased gut permeability.

Abstract:

Abstract A N-balance study was conducted to compare the effect of D-methionine (D-Met) or L-methionine (L-Met) supplementation on N balance, gut morphology and antioxidant status of weaned pigs. Fifty-six weaned barrows (10.5 ± 1.2 kg initial BW) were allotted to 7 diets in 2 blocks. A Met-deficient basal diet (BD; 0.24% standardized ileal digestible (SID) Met) but adequate in other AA, was supplemented with 3 graded levels (0.036, 0.072, and 0.108%) of D-Met or L-Met. After a 7-d adaptation, feces and urine were collected quantitatively for 5 d to determine N balance. At the completion of the experiment, blood samples were collected from all pigs. Pigs fed the BD and pigs fed the highest level of SID Met (0.34%) of both Met sources were euthanized and tissue samples from liver, kidney, muscle (longissimus dorsi), duodenal and jejunal mucosa were collected. N retention as % of N intake increased (P &lt; 0.001; 67, 72, 73, 74, 71, 74, 74%, respectively) by graded supplemental level of D-Met or L-Met. However, there was no interaction between Met source and supplemental level for all N-balance metrics. Using a slope-ratio regression, the bioavailability of D-Met relative to L-Met was 100.1% (95% confidence intervals: 85-116%) based on N retention (% of N intake). Villus height and crypt depth in the duodenum and jejunum were not affected by Met sources. Pigs fed the D-Met diet had a greater (P &lt; 0.05) total glutathione concentration in liver (4.9 vs. 1.5 µM) vs. BD. However, total antioxidant capacity and concentration of thiobarbituric acid-reactive substances in liver, muscle or plasma samples were not different among treatments. Supplementation with D-Met increased glutathione peroxidase activity in kidney (878 vs. 413 and 229 mU/mL; P &lt; 0.05) compared with BD or L-Met diet, however, activity of glutathione reductase in liver and kidney were not affected by treatments. These data indicate that D-Met and L-Met are equally efficient to support N retention, intestinal morphology and oxidative status in weaned pigs.

Abstract:

Unfixed cryostat sections of rat liver were incubated to demonstrate D-amino acid oxidase activity at the ultrastructural level. Incubation was performed by mounting the sections on a semipermeable membrane which was stretched over a gelled incubation medium containing D-proline as substrate and cerium ions as capture reagent for hydrogen peroxide. After an incubation period of 30 min, ultrastructural morphology was retained to such an extent that the final reaction product could be localized in peroxisomes, whereas the crystalline core remained unstained. Control incubations were performed in the absence of substrate; the lack of final reaction product in peroxisomes indicated the specificity of the reaction. We conclude that the semipermeable membrane technique opens new perspectives for localization of enzyme activities at the ultrastructural level without prior tissue fixation, thus enabling localization of the activity of soluble and/or labile enzymes.