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Effect of aminoglycoside otic drops on isolated cochlear outer hair cells with and without liver extract activation.

OBJECTIVE: To assess the ototoxicity of commercially available Gentacidin and TobraDex ear drops with and without liver extract activation using isolated cochlear outer hair cells (OHCs). MATERIAL AND METHODS: OHCs from adult chinchilla cochleae were exposed to standard bathing solution (SBS), liver extract alone and Gentacidin and TobraDex ear drops with and without liver extract. All experiments were performed at an osmolality of 305 +/-5 mOsm, at room temperature and for up to 60 min. OHC images were recorded using an inverted microscope and analyzed electronically. Time to cell death and changes in cell length were measured. RESULTS: The time to cell death and the percent change in cell length were significantly shorter in the Gentacidin+liver extract group than in the Gentacidin alone group (p < 0.05). The TobraDex+liver extract group showed a significantly decreased time to cell death compared to the SBS control group (p < 0.05). There were no significant differences in cell length or time to cell death between the TobraDex+liver extract group and the TobraDex alone group (p > 0.05). CONCLUSION: This study suggests that the cytotoxicity of aminoglycoside ear drops to isolated OHCs in vitro requires

Aminoglycosides↗

Soluble liver extract as supplement to detoxification by hemoperfusion in liver failure.

Charcoal hemoperfusion has been effective in reversing grade IV hepatic encephalopathy in patients. In animal studies essential factors have to be added to result in significant increase in the survival rate of grade III coma rats treated by hemoperfusion. In the present study, a water soluble liver extract was prepared and injected intraperitoneally into fulminant hepatic failure rats in grade II hepatic coma. This significantly (p less than 0.001) increased the survival time in these animals. However, no significant increase in survival rate was obtained. We were also able to significantly (p less than 0.0005) control gastrointestinal bleeding in the treated rats. We postulate that this supplementation of essential factors in combination with detoxification using hemoperfusion could form the basis of a more complete artificial liver support system.

Animals↗

Liver extract and its free amino acids equally stimulate gastric acid secretion.

Using intragastric titration in dogs with gastric fistulas, dose-response studies were carried out with liver extract and with a mixture of amino acids that matched the free amino acids found in liver extract. All solutions were adjusted to pH 7.0 and osmolality to 290 mosmol x kg-1. Doses are expressed as the sum of the concentrations of all free amino acids. At each dose studied (free amino acid concentration: 2.8, 5.6, 11, 23, and 45 mM), acid secretion in response to the free amino acid mixture was not significantly different from that of liver extract. The peak response to both liver extract and the free amino acid mixture occurred with the 23-mM dose and represented about 60% of the maximal response to histamine. The serum concentrations of gastrin after liver extract and the amino acid mixture were not significantly different. It is concluded that in dogs with gastric fistula, gastric acid secretion and release of gastrin were not significantly different in response to liver extract and to a mixture of amino acids that simulated the free amino acid content of liver extract.

Amino Acids↗

[Characteristic effects of rabbit liver extract on hepatocyte proliferation in the regenerating liver of mice of various ages].

Chalone isolated by the Verly method was injected into animals of different age (2 groups) under partial hepatectomy. The percentage of hepatocytes in S-, G2-, M- and post-M-periods was determined by morphoautoradiography. These parameters were used for the reconstruction of the extent and dynamics of cell proliferation during 36-48 hours of regeneration. The suppressing effect of chalone on hepatocyte proliferation was more pronounced in young mice. The lack of complete inhibition of DNA synthesis and of the effect of proliferative synchronization was observed in both groups of mice.

Aging↗

[Effect of the administration of liver extract after surgical operations in urology].

The effect of liver extract administered to 22 patients after urological operations was evaluated with reference to 19 controls. No significant difference was observed between the cases and the controls. We rather supposed, however, that the liver extract was effective as a liver protection drug, because the results of liver function in the cases were less changed than the controls throughout the post-operative course.

Adult↗

Double-blind study of a total liver extract in patients with hepatic dysfunction.

Forty hospitalized male patients with hepatic functional deficit were treated i.m. for 3 weeks with a total liver extract or placebo. The study was a double-blind. Statistical analysis (Student's t test, analysis of variance. Mann-Whitney test and Fisher's probability test) showed that liver extract therapy was more effective than placebo in improving liver function as indicated by changes in clinical features(digestive disorders, constipation, hepatomegaly) and colloidal and other laboratory tests, e.g. plasma cholesterol, prothrombin time, red cell count and hemoglobin concentration. An overall clinical assessment showed that 60% of patients treated with the liver extract were improved. In addition, no local or general side effects were observed. Present results are in favour of an hepatoprotective activity of a total liver extract.

Adult↗

The intestinal phase of gastric secretion. Response to liver extract infusion into the proximal jejunum of healthy human subjects.

A Levine tube was placed under radiological control in the stomach, and a thin polyethylene tube in the proximal jejunum of 6 healthy volunteers. The stomach and proximal part of jejunum were perfused for 2 hours with 1% acetylcholine, 20% meat extract (Bovril), and 15% liver extract (LE) alone and in combination with simultaneous infusion of different doses of exogenous pentagastrin intravenously. A significant increase in serum gastrin concentration was found with antral perfusion of LE only, whereas perfusion of the proximal jejunum did not change the basal level of the serum gastrin concentration. No change from control values was observed in gastric acid, and pepsin output on perfusing proximal jejunum with LE alone, or in combination with pentagastrin. Reflux to the stomach varied between 0-1.4%, as determined by addition of radioactive B12 to the perfusates. The experiments showed that gastrin was released from the antrum of the stomach by perfusion with 15 per cent LE, but not from the jejunum under the present experimental conditions. In the present experiments Bovril and acetylcholine perfusions did not cause significant responses from the antrum or from the proximal jejunum.

Acetylcholine↗

Plasma secretion and pancreatic secretion in response to liver extract meal with varied pH and exogenous secretin in the dog.

1. In dogs with chronic gastric and pancreatic fistulas, a liver extract meal adjusted to various pH levels ranging from 7.0 to 2.0, was introduced into the stomach and the increments in plasma secretin levels were correlated with the pH of the liver extract meal and pancreatic bicarbonate outputs. 2. The pH threshold for both bicarbonate secretion and secretin release was found to be about 4.5. With a stepwise decrease in the pH of the meal below pH 4.5, there were stepwise increments in the plasma secretin concentrations and pancreatic bicarbonate outputs. 3. Exogenous secretin, given in graded doses ranging from 0.03 to 2.0 clinical unit/kg per hr, increased the plasma secretin concentrations and bicarbonate secretion in a dose-dependent fashion. 4. These results indicate that the pH threshold for release of endogenous secretin is 4.5 and suggests that, at pH levels below 4.5, pancreatic bicarbonate secretion depends upon the duodenal acid load and is linearly correlated to an increment in plasma secretin concentrations. 5. It is concluded that endogenous secretin is a major determinant of pancreatic bicarbonate secretion after a meal. 6. Pancreatic protein secretion by intragastric liver extract meal was greatly increased both in experiments with liver extract meal, pH 4.0 or below, and I.V. infusion of secretin at a dose of 0.12 u./kg per hr. It is questioned, however, whether this effect of secretin is physiological.

Animals↗

Inhibition of protein and nucleic acid synthesis of animal cells in vitro mediated by high molecular weight inhibitors in human liver extract.

1. The addition of human liver extract to HeLa cells induces a reversible inhibition of the incorporation of [3H] thymidine into the DNA, [3H] uridine into the RNA, and 14C-labelled amino acids into the protein of HeLa cells. The inhibitory effects appear after treatment for 1 h and reach a maximum after 4-8 h. These effects do not depend on a defective precursor penetration, isotopic dilution or degradation of labelled precursor (thymidine-degrading enzymes were inactivated by the addition of unlabelled thymine), reduced activity of thymidine and uridine kinase, medium impairment, or an impairment of the cell-membrane function. 2. The nucleic acid synthesis-inhibiting activity of the extract seems to be dependent on cellular protein synthesis but independent of RNA synthesis which indicates that the inhibitors act in an indirect way. Furthermore, the inhibitors seem to lack the tissue-specific character of chalones. 3. The extract contains separate inhibitors of DNA, RNA and protein synthesis. These inhibitors were found to have different physical-chemical characteristics and to be macromolecules with a protein or conjugated protein character (mol. wt. approx. 90 000). 4. The possibility that the activity of the high molecular weight inhibitors resides in low molecular weight factors (bound to protein carriers) was tested: No true low molecular weight inhibitors could be liberated by extraction with trichloroacetic acid/organic solvents or by dialysis/enzymatic treatments. Nucleosides such as thymidine, uridine, and cytidine, however, were liberated and could be shown to interfere with the uptake of [3H] thymidine/[3H] uridine.

Cell Line↗

Amelioration of non-alcoholic steatohepatitis and glucose intolerance in ob/ob mice by oral immune regulation towards liver-extracted proteins is associated with elevated intrahepatic NKT lymphocytes and serum IL-10 levels.

Non-alcoholic steatohepatitis (NASH) is a common cause of cryptogenic cirrhosis in the Western world. In an animal model of NASH, leptin-deficient ob/ob mice present with alterations in number and function of hepatic NKT and peripheral CD4 lymphocytes. Oral immune regulation is a method to alter the immune response towards orally administered antigens. To determine the effect of oral immune regulation towards liver-extracted proteins on the metabolic disorders in ob/ob mice, ob/ob mice and their lean littermates were orally administered liver extracts from wild-type or ob/ob mice or bovine serum albumin for 1 month. The effect of treatment on hepatic fat content was measured by magnetic resonance imaging (MRI) and using a histological steatohepatitis grading scale. Glucose tolerance was measured by an oral glucose tolerance test (GTT). T lymphocyte subpopulations were assessed by flow cytometry analysis. Induction of immune regulation by oral presentation of liver-extracted proteins resulted in a significant 18% reduction of the hepatic fat content in ob/ob mice fed with either wild-type or ob/ob liver extracts for 1 month. The MRI signal intensity index in treated mice decreased to 0.48 and 0.51, respectively, compared with 0.62 in BSA-fed controls (p = 0.037 and p = 0.019, respectively), while the histological steatohepatitis score decreased in both treated groups to 2.0, compared with 2.4 in BSA-fed controls (p = 0.05). A significant improvement in GTT was noted in treated ob/ob mice. These changes were accompanied by a marked increase in the intrahepatic NKT lymphocyte population in mice fed with proteins extracted from both wild-type and ob/ob mice (46.96% and 56.72%, respectively, compared with 26.21% in BSA-fed controls; p < 0.05) and a significant elevation in serum IL-10 levels. Oral immune regulation towards liver extracted proteins in leptin-deficient mice resulted in a marked reduction in hepatic fat content and improved glucose tolerance. This effect was associated with a significant increase in the intrahepatic NKT lymphocyte population and serum IL-10 levels, suggesting a Th1 to Th2 immune shift. Immune regulation towards disease-associated antigens holds promise as a new mode of therapy for NASH.

Adipose Tissue↗