The effect of grape seed and grape marc meal extract on milk performance and the expression of genes of endoplasmic reticulum stress and inflammation in the liver of dairy cows in early lactation.

During the periparturient phase, cows are typically in an inflammation-like condition, and it has been suggested that inflammation associated with the development of stress of the endoplasmic reticulum (ER) in the liver contributes to the development of fatty liver syndrome and ketosis. In the present study, we investigated the hypothesis that feeding grape seed and grape marc meal extract (GSGME) as a plant extract rich in flavonoids attenuates inflammation and ER stress in the liver of dairy cows. Two groups of cows received either a total mixed ration as a control diet or the same total mixed ration supplemented with 1% of GSGME over the period from wk 3 prepartum to wk 9 postpartum. Dry matter intake during wk 3 to 9 postpartum was not different between the 2 groups. However, the cows fed the diet supplemented with GSGME had an increased milk yield and an increased daily milk protein yield. Cows supplemented with GSGME moreover had a significantly reduced mRNA abundancy of fibroblast growth factor (FGF) 21, a stress hormone induced by various stress conditions, in the liver in wk 1 and 3 postpartum. In contrast, mRNA abundances of a total of 3 genes involved in inflammation and 14 genes involved in ER stress response, as well as concentrations of triacylglycerols and cholesterol, in liver samples of wk 1 and 3 postpartum did not differ between the 2 groups. Overall, this study shows that supplementation of GSGME did not influence inflammation or ER stress in the liver but increased milk yield, an effect that could be due to effects on ruminal metabolism.

Association of Doberman hepatitis to canine major histocompatibility complex II.

Doberman hepatitis (DH) is a chronic and progressive inflammatory liver disease that mainly affects female dogs. The high incidence of chronic hepatitis in Dobermans is suggestive of a genetic predisposition. DH is characterized by mononuclear cell infiltration and copper accumulation in the liver and major histocompatibility complex (MHC) class II antigen expression in the hepatocytes. In dogs, the MHC is referred to as the dog leukocyte antigen (DLA) system. In this study, the potential role of DLA genes in DH was investigated by sequence-based typing in the exon 2 of DLA-DRB1, -DQA1 and -DQB1. The case group comprised 37 Dobermans with subclinical or clinical DH. The control group consisted of 37 healthy Dobermans, with normal liver enzyme values and without immunosuppressive medication. The control dogs were over 10 years old to include dogs with the lowest genetic risk of DH. Our results indicate that Dobermans with homozygous DLA-DRB1*00601/DQA1*00401/DQB1*01303 [odds ratio (OR) = 14.9, confidence limit (CL) = 3.1-71.7, P < 0.00005], especially with homozygosity for DLA-DRB1*00601 (P < 0.0005), are susceptible to DH. The DQ heterodimer DLA-DQA1*00901/DQB1*00101 and the allele DLA-DRB1*01501 appear to confer protection against DH (P < 0.001). Allele and haplotype frequencies were compared using chi-squared statistics. The disease shows a complex pattern of inheritance, but the observed DLA class II association with DH suggests a role for the immune system in the development of the disease.

Adenoviral hepatitis in two Nile crocodile (Crocodylus niloticus) hatchlings from South Africa.

Adenoviral infections may cause mild to severe morbidity or fatality in a large array of animal species. In crocodilians, hatchlings under 5 months of age are usually affected. However, there is a paucity of information on actual incidences in hatchlings originating from South Africa. Two cases of adenoviral hepatitis in crocodile hatchlings about 2 weeks old, bred on a commercial farm in South Africa, are described. Both hatchlings showed typical clinical signs of hepatitis. The identification of intranuclear inclusion bodies in the liver was used to differentiate between adenoviral hepatitis and chlamydial hepatitis. Although vertical transmission has never been proven in crocodiles, the young age of the affected hatchlings raises the possibility of vertical transmission. The lack of epidemiological information on adenoviral hepatitis in crocodiles highlights the need for further characterisation of the virus and targeted surveillance.

Inflammatory stress and altered angiogenesis evoked by very high-fat diets in mouse liver.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD), a condition that leads to fibrosis, is caused by intake of very high-fat diets (HFDs). However, while the negative impact on the liver of these diets has been an issue of interest, systematic research on the effect of HFDs are lacking. OBJECTIVE: To characterize the overall impact of HFDs on both molecular and morphological signs of liver remodeling. METHODS: A study was conducted on male C57BL/6J mice to assess the effect of 4- and 8-week HFDs (60% kcal from fat) on (i) liver steatosis and fibrosis, and (ii) expression of factors involved in inflammation and angiogenesis. RESULTS: After an 8-week HFD, vascular endothelial growth factor type-2 receptor (VEGF-R2) and fatty acid translocase/trombospondin-1 receptor (CD36) were overexpressed in liver tissue of mice given HFDs. These changes suggest impaired liver angiogenesis and occurred together with (i) increased GPR78-BiP and EIF2α phosphorylation, suggesting endoplasmic reticulum stress, (ii) induction of Col1a1 gene expression, a marker of fibrosis, and (iii) increased CD31 immunolabeling, consistent with active angiogenesis and fibrosis. CONCLUSION: Our data show that very HFDs promote a rapid inflammatory response, as well as deregulation of angiogenesis, both consistent with development of liver fibrosis.

Canine Copper-Associated Hepatitis.

Copper-associated hepatitis is recognized with increasing frequency in dogs. The disease is characterized by centrolobular hepatic copper accumulation, leading to hepatitis and eventually cirrhosis. The only way to establish the diagnosis is by histologic assessment of copper distribution and copper quantification in a liver biopsy. Treatment with the copper chelator d-penicillamine is the most commonly used treatment. In addition, a low-copper/high-zinc diet can help prevent accumulation or reaccumulation of hepatic copper. Mutations in the copper metabolism genes COMMD1 or ATP7A and ATP7B have been associated with hepatic copper concentrations in Bedlington terriers and Labrador retrievers respectively. In the Labrador retriever, dietary copper intake contributes strongly to the disease phenotype.

Canine Idiopathic Chronic Hepatitis.

The World Small Animal Veterinary Association's Liver Standardization Group produced standardized criteria for the histologic diagnosis of canine chronic hepatitis (CH). They define CH by the presence of hepatocellular apoptosis or necrosis, a variable mononuclear or mixed inflammatory cell infiltrate, regeneration, and fibrosis. There are variations in histologic appearance between breeds. Hepatic copper accumulation is an important cause of canine CH. However, where copper accumulation has been ruled out, dogs are said to have idiopathic CH. This article reviews theories regarding the etiopathogenesis of canine CH other than copper accumulation, and its clinical features, diagnostic findings, and management.

Washout Ratio in the Hepatic Vein Measured by Contrast-Enhanced Ultrasonography to Distinguish Between Inflammatory and Noninflammatory Hepatic Disorders in Dogs.

BACKGROUND: Perflubutane microbubbles, a second-generation ultrasound contrast agent, are phagocytized by Kupffer cells. This characteristic may be useful to differentiate diffuse hepatic diseases in dogs. HYPOTHESIS/OBJECTIVES: To determine whether the washout ratio in the hepatic vein (HV) measured by contrast-enhanced ultrasonography (CEUS) can distinguish between inflammatory and noninflammatory hepatic disorders in dogs. ANIMALS: Forty-one client-owned dogs with hepatic disorders including 14 with hepatitis, 7 with primary hypoplasia of the portal vein (PHPV), 9 with congenital portosystemic shunt (cPSS), and 11 with other hepatopathy were enrolled. Six dogs without hepatic disease also were evaluated as healthy controls. METHODS: Dogs with hepatic disorders were prospectively included. Contrast-enhanced ultrasonography of the HV was performed for 2 minutes. Washout ratio was defined as the attenuation rate from peak intensity to the intensity at the end of the CEUS study. RESULTS: Washout ratio in the hepatitis group (median, 18.0%; range, 2.0-37.0%) was significantly lower than that of the PHPV (median, 52.2%; range, 11.5-86.3%), cPSS (median, 60.0%; range, 28.6-77.4%), other hepatopathy (median, 70.5%; range, 26.6-88.4%), and normal (median, 78.0%; range, 60.7-91.7%) groups. The area under the receiver operating characteristic curve for hepatitis was 0.960, with a 95% confidence interval (CI) of 0.853-0.990. Washout ratio ≤37.1% resulted in a sensitivity of 100% (95% CI, 78.5-100%) and specificity of 85.2% (95% CI, 67.5-94.1%) for the prediction of hepatitis. CONCLUSIONS AND CLINICAL IMPORTANCE: Washout ratio can distinguish hepatitis from the other noninflammatory disorders with high accuracy. This result might reflect impaired Kupffer cell phagocytosis in dogs with hepatitis.

Novel insights into the biology and physiology of the Ito cell.

Ito cells, perisinusoidal mesenchymal elements with possible pericytic functions within the liver, recently have been shown to play multiple physiological and pathophysiological roles. In particular, several in vivo and in vitro studies have clearly indicated that Ito cells play a relevant role in the progression of liver fibrogenesis. More recently, attention has been focussed on the mechanisms leading to Ito cell activation, proliferation and synthesis of extracellular matrix components. Among other soluble factors potentially involved in these processes, transforming growth factor-beta 1 and platelet-derived growth factor have been shown to act in a paracrine, and possibly autocrine, fashion on Ito cells, thus perpetuating their activated state. Finally, other studies have shown that Ito cells could play an active role in chronic liver tissue inflammation by promoting chemotaxis of infiltrating inflammatory cells.

Requirement of peptidergic sensory innervation for disease activity in murine models of immune hepatitis and protection by beta-adrenergic stimulation.

To investigate the interaction between the peripheral nervous and the immune system in vivo, we used two mouse models of T cell and TNF-alpha dependent liver injury inducible by either concanavalin A or a combination of D-galactosamine and staphylococcal enterotoxin B. Mice depleted of peptidergic sensory nerve fibres by capsaicin were protected from liver injury. Moreover, TNF-alpha production was significantly reduced. Examination of the effect of catecholamines on liver injury showed that the beta2-adrenergic agonist salbutamol prevented, whereas chemical sympathectomy by 6-hydroxydopamine, deteriorated the disease. Hence, strategies reducing the activity of peptidergic sensory nerve fibres or stimulating beta2-adrenoreceptors, may be of benefit in immune-mediated liver disease.

Distinct effects of systemic infusion of G-CSF vs. IL-6 on lung and liver inflammation and injury in hemorrhagic shock.

Production of pro-inflammatory cytokines, such as granulocyte colony-stimulating factor (G-CSF) and interleukin-6 (IL-6) occurs at multiple tissue sites in hemorrhagic shock (HS), resulting in elevated circulating plasma levels. The current study was designed to test the hypothesis that circulating G-CSF and IL-6 contribute to polymorphonuclear neutrophilic granulocyte (PMN)-mediated inflammation and organ injury in HS. Sprague-Dawley rats were subjected to decompensated HS (mean arterial blood pressure = 40 mm Hg for 2.5 h), followed by resuscitation with lactated Ringer's solution with or without G-CSF (3 microg/kg) or IL-6 (3 microg/kg). Animals were killed 4 h after resuscitation, and their lungs and livers were assessed quantitatively for PMN infiltration, organ injury, and activation of NF-kappaB and signal transducer and activator or transcription (STAT) 3. Infusion of G-CSF during resuscitation increased PMN infiltration into the lungs by 2.4-fold (P < 0.01) compared with animals resuscitated with lactated Ringer's solution alone. Increased PMN infiltration was accompanied by interstitial edema and pneumocyte swelling, resulting in a 42% increase in lung alveolar wall cross-sectional surface area (P < 0.01) and a 3.7-fold increase in Stat3 activity (P < 0.01). G-CSF infusion did not affect PMN infiltration into the liver and was accompanied by a 68% decrease in focal hepatocellular necrosis (P < 0.01). Infusion of IL-6, in contrast, dramatically decreased inflammation and injury in both the lung and liver; the anti-inflammatory effects of IL-6 may be mediated, in part, by down-modulation of nuclear factor (NF)-kappaB activity. Thus, circulating G-CSF and IL-6 have opposing effects on PMN recruitment and injury in the lung in HS while both protect against hepatic necrosis. The beneficial effect of these cytokines on liver injury in HS appears to be independent of PMN recruitment.

Experimental reproduction of Escherichia coli cellulitis and septicemia in broiler chickens.

Experimental reproduction of avian cellulitis was conducted by subcutaneous inoculation of 25-day-old broiler chickens with a field isolate of serogroup O78 Escherichia coli. Development of the cellulitis lesion occurred as early as 24 h post-infection. Reproduction of cellulitis occurred in 98% of inoculated birds, and E. coli was isolated from > 75% of cellulitis lesions. In addition to cellulitis, other lesions, including pericarditis, airsacculitis, osteomyelitis, arthritis, and perihepatitis, occurred in > 80% of birds inoculated with E. coli. Bacteremia occurred as early as 6 h post-infection and dramatically declined by 5 days post-infection. Seventeen of 59 (29%) birds inoculated with E. coli developed a fatal infection between 1 and 6 days post-infection, and bacteria were isolated from lesions in 98% birds. In contrast, E. coli was not isolated from lesions in birds that survived until days 7-14 post-infection. Birds that survived with cellulitis and other lesions until day 14 post-infection had a significantly lower body weight compared with the control group. This avian model of cellulitis and other lesions will be useful for studying the development of vaccination strategies for E. coli in broilers.

Age-dependent phagocytosis of erythrocytes by the isolated perfused rat liver after galactosamine hepatitis and alpha-naphthylisothiocyanate cholestasis.

UNLABELLED: Isolated rat livers were perfused with an immunoglobulin-free fluid containing homologous rat erythrocytes suspended in an isotonic saline solution. Galactosamine hepatitis and alpha-naphthylisothiocyanate cholestasis were induced as experimental liver diseases. The glutamic oxalacetic transaminase (GOT) and guanosine triphosphate (GPT) activities, the potassium level as well as the redox quotients of lactate/pyruvate and beta-hydroxybutyrate/acetoacetate were determined as liver function parameters. RESULTS: The same dose of galactosamine led to two different types of reaction. The group suffering more damage (rendering maximum GOT activities) phagocytosed significantly (p less than or equal to 0.05) more erythrocytes than the other group. Galactosamine hepatitis significantly slows down the phagocytosis of erythrocytes. The function of the mononuclear phagocytosing cells in the liver is intact. The alpha-naphthylisothiocyanate (ANIT) cholestasis significantly reduces the capacity of the mononuclear phagocytosing cells in the liver. Young erythrocytes were phagocytosed significantly better than old ones in either type of liver damage, in galactosamine hepatitis and in ANIT cholestasis as well as by healthy livers.

Experimental hepatitis induced by Campylobacter jejuni infection in Japanese quail (Coturnix coturnix japonica).

To establish an experimental model for vibrionic hepatitis caused by Campylobacter jejuni, Japanese quails (Coturnix coturnix japonica) were inoculated with C. jejuni strains isolated from chicken hepatitis (BL107) and human diarrhea (HP5113). Necrotic liver lesions were formed by intra-pancreaticoduodenal vein injection by which the bacteria reached the liver directly via the portal vein, but not by intra-gastric infection. These liver lesions were observed from day 1 to 7 after the infection. The pathological changes were weak and no clinical signs were observed throughout the experimental period. By immunohistochemistry, the bacterial antigens were detected in the hepatocytes, and intercellular spaces between the hepatocytes, and in the macrophages during the early stage of the infection. When focal hepatocyte necrosis was formed, the antigen was detected more frequently in the intact hepatocytes at the periphery of the lesions than within necrotic foci. The bacteria were not detected from the liver, spleen or blood according to raising the serum agglutination titer. In contrast, the bacteria immediately invaded the bile in 5 min post-infection and were retained in the gallbladder for long periods. The present study showed that necrotizing hepatitis was formed by intra-pancreaticoduodenal vein infection of the quail with C. jejuni.

[Adaptive changes in the body upon exposure to electromagnetic radiation]. / Adaptivnye izmeneniia v organizme pri deistvii élektromagnitnykh izluchenii.

The chance to use electromagnetic exposures as active adaptogen and the detecting of adaptive changes following them were objects of our studies. The data of experimental and clinical studies significative the dependence of changes on the functional state of organism were seen. Particular attention is paid to the site of exposure and to the advantages in the action of electromagnetic exposures on areas overlaying the endocrine glands and control centers of central nerve system. In these conditions electromagnetic exposures play a part of trigger initiated natural processes of homeostatic regulation in the organism functional systems. It is shown that the course of electromagnetic exposures in wide frequency range until laser radiation (infrared and red) arises adaptive changes of the regulator systems, of the bioenergetic and the biosynthetic processes in myocardium, liver, brain, thymus and other tissues predetermined genetically and secured the power of the adaptive systems. The cross-adaptation effects underlie the electromagnetic exposures medical action.

[Cytokine network involved in inflammatory reactions].

A cytokine, present in a minute amount in vivo, generally exhibits multiple functions shared by other cytokines. The recent progress in molecular biology and protein chemistry have facilitated identification of the structures and functions of cytokines and their receptors. This has resulted to the clinical application of several cytokines, such as granulocyte colony-stimulating factor and erythropoietin. However, the elucidation of the pathophysiological roles of cytokines in various diseases, has been hindered by a complicated network where cytokines regulate the production and/or functions of others. The utilization of homologous recombination and ES cells has led to the generation of mice that are deficient in a particular cytokine or cytokine receptor gene. Analyses of these deficient mice have given us invaluable information to unravel the roles of each cytokine or cytokine receptor. Here, we described our studies on the pathophysiological roles of pro-inflammatory cytokines using cytokine or cytokine receptor gene-deficient mice to provide examples of the usefulness of these mice in clarifying the cytokine network.

[Investigation of the hepatoprotector action of natural flavonoids]. / Izuchenie gepatozashchitnogo deistviia prirodnikh flavonoidnykh soedeninii.

The prophylactic hepatoprotective properties of a series of natural flavonoids (hesperidin from citrus peel, diosmine from Vicia tanuifolia Roth., and diosmine analog from Vicia trunctula) were studied in a group of white male rats with a model of acute CCl4 hepatitis. The effect was evaluated by the ability of tested substances to normalize the biochemical characteristics of the functional state of the liver, in comparison to the reference drug carsil. The most pronounced hepatoprotective action was observed for diosmine analog administered in a dose of 100 mg/kg, which was superior in some respects to the reference hepatoprotector (in equivalent doses). This is probably related to the higher bioaccessibility of diosmine analog ensured by a carbohydrate moiety of this compound, in combination with the C2-C3 double bond present in diosmetin (diosmin aglycon).

[Effect of antihepatotoxic tea on the course of drug-induced hepatitis]. / Vliianie antigepatotoksicheskogo chaia na techenie lekarstvennogo gepatita.

Pharmacotherapeutic efficacy of antihepatoxic tea was studied on an experimental model of tetracycline-induced hepatitis. It was concluded that the tea had a marked pharmacotherapeutic effect on the process of tetracycline-induced hepatitis in animals. It lowered the level of the cytolytic syndrome, prevented the progress of cholestasis and stimulated the bile secretory function of the liver. The favourable effect of the plant antihepatotoxic preparation was due to the presence of a complex of its biologically active substances.

[Effect of maternal toxic hepatitis on the functional characteristics of the lactation process]. / Vliianie toksicheskogo gepatita u materi na funktsional'nye osobennosti protsessa laktatsii.

The authors have studied the influence of chronic heliotrine intoxication on female rats subjected to it before their pregnancy and on the quantity of proteins, carbohydrates, activity of enzymes and immunocompetent cells (ICC) of rats milk during the period of breast feeding. Decrease in an amount of proteins and carbohydrates since the third day of lactation, lowering in dipeptidhydrolase, r-amilase and maltase activity were observed in the study. It has been also seen the decrease in an amount of ICC (monocytes, macrophages, small lymphocytes) just after parturitions. This deacrease was mostly expressed on the 14th day of lactation. IC cells were not determined in milk on the 21st day of breast feeding.

[Expression of hepatocyte growth factor (HGF) in LEC rats at various phases of hepatitis and hepatoma].

The role of hepatocyte growth factor (HGF) in LEC rats were investigated at various phases of liver diseases by the detection of HGF expression, using ELISA assay, mRNA analysis and immunohistochemistry. Levels of plasma HGF increase in the fulminant hepatitis phase, decreased during chronic/cholangiofibrosis phase, and in some LEC rats, high levels of HGF were observed in hepatoma phase. HGF mRNA level in the liver was very high in fulminant hepatitis phase and low in chronic hepatitis phase. In hepatoma phase, HGF mRNA level was intermediate or high in the liver. In fulminant hepatitis phase, HGF mRNA level in the lung was slightly increased, while it was almost stable in the kidney in all the conditions studied. Immunohistochemical studies revealed that the frequency of HGF positive cells increased remarkably in fulminant hepatitis phase, and that many of them were located at the portal triads. Fewer HGF-positive cells were found in chronic hepatitis phase and were not found in the tissue of cholangiofibrosis. HGF was found in the surface of hepatocellular carcinoma cells and in the cytoplasm of the non-epithelial cells in cancerous liver tissues. HGF-positive cells appeared 24h after partial hepatectomy, diffusely, and HGF mRNA increased earlier in the kidney and lung than in the liver. Moreover, HGF mRNA level was higher in the lung than in the liver. These results suggest that in the natural course of spontaneous hepatitis and hepatoma in LEC rats, HGF is expressed mainly in the liver and that HGF may play an important role in the regeneration of hepatocytes in a paracrine manner. In contrast, after partial hepatectomy, HGF produced in the lung may be effective for liver regeneration in an endocrine manner.

Assessment of hepatic inflammation after spinal cord injury using intravital microscopy.

OBJECTIVES: The liver has been shown to play a particularly important role in the initiation and progression of the early systemic inflammatory response (SIR) to spinal cord injury (SCI). The purpose of this study was to determine the time course of leucocyte recruitment to the liver, and to determine the effect of injury severity on the magnitude of leucocyte recruitment and hepatic injury. METHODS: Rats were randomly assigned to one of the following groups: uninjured, sham-injured (laminectomy and no cord injury), cord compressed or cord transected. At 30 min and 90 min after SCI rats had the left lobe of their livers externalised and visualised using intravital video microscopy. RESULTS: Thirty minutes after injury the total number of leucocytes per post-sinusoidal venule was significantly increased after cord transection compared to that in uninjured and sham-injured rats (P<0.05). Of these leucocytes, significantly more were adherent to venule walls (P<0.05). At 90 min the total number of leucocytes per post-sinusoidal venule and the number of adherent and rolling leucocytes was significantly increased after cord transection and cord compression (P<0.05). DISCUSSION: This is the first study to use intravital microscopy to visualise systemic inflammation in the liver following SCI. We have demonstrated immediate leucocyte recruitment to the liver within 30 min after injury and have shown that systemic inflammation increases with time after injury and with severity of injury.