Intraocular pressure in cats is lowered by drops of hornet venom

1. Nine cats were given an intravenous injection of the Oriental hornet (Vespa orientalis, Vespinae; Hymenoptera) venom sac extract (VSE) and seven cats had the same VSE administered as eye drops. 2. When injected intravenously, the hornet VSE decreased the intraocular pressure in both eyes sharply during the first 20 min and with a slower rate later on until the end of the 3 hr experiment. The intraocular pressure dropped to zero in some cases. 3. VSE eye drops decreased the intraocular pressure only in the treated eye, while in the second eye (left as a control) the intraocular pressure remained the same throughout the experiment. 4. The decrease in the intraocular pressure was sharp during the first 20 min and slowed down afterwards until the end of the experiment. 5. The intraocular pressure did not reduce to zero. 6. This study shows that the active components of the hornet venom which caused a decrease in the intraocular pressure can cross the cornea and exert a hypotensive effect in the eye.     

Cardioactive effects of hornet venom, Vespa mandarinia

In the experiment of electrocardiogram, the crude venom of giant hornet (Vespa mandarinia) showed cardioactive effects on rat heart. The heart rate was accelerated within 5 min after injection of the venom intraperitoneally, then the heart beat was blocked, resulting in conduction delay. The cardioactive constituent was separated into two components by gel filtration. One which was high molecular species such as protein showed complete atrioventricular block. Another component, having a low molecular weight, was fractionated in 5 peaks, which accelerated heart rate.     

Bile acids secretion and synthesis by isolated rat hepatocytes.

Bile acids secretion and their distribution were studied in isolated rat hepatocytes. Bile acids secretion was linearly related with time for first three hours of incubation and the net secretion rate was 23.2 ± 2.74 nmoles per g cells (wet weight) per minute. Isolated hepatocytes synthesized relatively more chenodeoxycholic acid than cholic acid compared to whole animal. These results suggest that isolated hepatocytes synthesize and secrete bile acids and thus provide experimental system to study the effect of drugs on bile acids secretion and synthesis at cellular level.     

Bile acid sulfates in serum bile acids determination.

Some bile acid sulfates were synthesized and characterized. The configuration of sulfate groups at C-3, C-7 and C-12 positions was confirmed by Nuclear Magnetic Resonance analysis. These sulfates were utilized in a study of their chemical behaviour in different analytical procedures currently used for serum bile acids determination. Procedures for bile acids extraction from serum with ethanol or Amberlite XAD-2 result in an important loss of the most polar sulfated bile acids. Complete separation of unsulfated from sulfated bile acids on Sephadex LH-20 is not achieved when deconjugation of the most polar bile acid sulfate is slow but does not produce artifacts. Enzymatic determination of bile acids gives positive response with some bile acid sulfates. The current procedures of serum bile acids determination are discussed in consideration of these results.     

Bile acids as carcinogens in human gastrointestinal cancers

Bile acids were first proposed to be carcinogens in 1939 and 1940. On the basis of later work with rodent models, bile acids came to be regarded as cancer promoters rather than carcinogens. However, considerable indirect evidence, obtained more recently, supports the view that bile acids are carcinogens in humans. At least 15 reports, from 1980 through 2003, indicate that bile acids cause DNA damage. The mechanism is probably indirect, involving induction of oxidative stress and production of reactive oxygen species that then damage DNA. Repeated DNA damage likely increases the mutation rate, including the mutation rate of tumor suppressor genes and oncogenes. Additional reports, from 1994 through 2002, indicate that bile acids, at the increased concentrations accompanying a high fat diet, induce frequent apoptosis. Those cells within the exposed population with reduced apoptosis capability tend to survive and selectively proliferate. That bile acids cause DNA damage and may select for apoptosis-resistant cells (both leading to increased mutation), indicates that bile acids are likely carcinogens. In humans, an increased incidence of cancer of the laryngopharyngeal tract, esophagus, stomach, pancreas, the small intestine (near the Ampulla of Vater) and the colon are associated with high levels of bile acids. The much larger number of cell generations in the colonic (and, likely, other gastrointestinal) epithelia of humans compared to rodents may allow time for induction and selection of mutations leading to cancer in humans, although not in rodents.     

Bile acids inhibit secretion of very low density lipoprotein by rat hepatocytes

The influence of taurocholate on very low density lipoprotein (VLDL) triacylglycerol synthesis and secretion was studied by isolated rat liver-parenchymal cells. The incorporation of [3H]glycerol into cell-associated and VLDL triacylglycerols were measured after incubation in medium containing 0.75 mM oleate. Taurocholate caused a maked decrease in VLDL [3H]triacylglycerol secretion from the hepatocytes: 50-150 microM taurocholate inhibited secretion of VLDL [3H]triacylglycerols by 70-90%. Similar results were obtained when the mass of secreted VLDL triacylglycerols was measured. Taurocholate caused a decreased secretion of VLDL [3H]triacylglycerols after 15-30 min incubation. A higher amount of cellular triacylglycerols was found in taurocholate-supplemented cells. Furthermore taurocholate did not change the intracellular lipolysis of triacylglycerols. These results suggest that bile acids interfere more probably with the assembly and/or secretion of VLDL-particles and not with earlier stages of VLDL formation, e.g. triacylglycerol synthesis.     

Cadmium-induced hepatotoxicity in cultured rat hepatocytes as evaluated by morphometric analysis

Freshly isolated hepatocytes from neonatal rats were cultured for approximately 24 h; incubated for 5, 30, or 60 min in solutions containing 0, 50, 100, or 200 microM cadmium; embedded in plastic; and sectioned for optical microscopy. The extent of cadmium-induced hepatotoxicity was evaluated by double-blind morphometric analysis (a geometricostatistical processing of two-dimensional data for the collection of three-dimensional information) whereby hepatocytes were classified on the basis of the severity of morphologic damage at the optical level. Both time and concentration effects were studied. Cultures exposed to 200 microM cadmium, for various intervals of time from 5 to 60 min, showed statistically significant reductions in the relative volume percent of normal hepatocytes, elevations (then reductions) in the relative volume percent of slightly damaged hepatocytes, increases in the relative volume percent of moderately damaged cells, and increases in the relative volume percent of severely damaged liver cells. As the concentration of cadmium was increased from 50 to 200 microM cadmium (during both 30 and 60-min exposures), significant trends were observed in cellular distribution patterns based on relative volume percent. Morphologically normal cells decreased, both slightly damaged and moderately damaged cells increased, and severely damaged cells remained unchanged. These results indicated that morphometric analysis at the optical level provided quantitative estimates for the evaluation of time- and concentration-effects of cadmium on cultured hepatocytes.     

Cadmium-induced hepatotoxicity in cultured rat hepatocytes as evaluated by morphometric analysis

Summary Freshly isolated hepatocytes from neonatal rats were cultured for approximately 24 h; incubated for 5, 30, or 60 min in solutions containing 0, 50, 100, or 200 μM cadmium; embedded in plastic; and sectioned for optical microscopy. The exeent of cadmium-induced hepatotoxicity was evaluated by double-blind morphometric analysis (a geometricostatistical processing of two-dimensional data for the collection of threedimensional information) whereby hepatocytes were classified on the basis of the severity of morphologic damage at the optical level. Both time and concentration effects were studied. Cultures exposed to 200 μM cadmium, for various intervals of time from 5 to 60 min, showed statistically significant reductions in the relative volume percent of normal hepatocytes, elevations (then reductions) in the relative volume percent of slightly damaged hepatocytes, increases in the relative volume percent of moderately damaged cells, and increases in the relative volume percent of severely damaged liver cells. As the concentration of cadmium was increased from 50 to 200 μM cadmium (during both 30 and 60-min exposures), significant trends were observed in cellular distribution patterns based on relative volume percent. Morphologically normal cells decreased, both slightly damaged and moderately damaged cells increased, and severely damaged cells remained unchanged. These results indicated that morphometric analysis at the optical level provided quantitative estimates for the evaluation of time- and concentration-effects of cadmium on cultured hepatocytes. This work was supported by a grant from the Johns Hopkins University Center for Alternatives to Animal Testing.     

A fatal effect of hornet venom on rat-brain cortical neurons

In a previous study, it was shown that the hornet venom or, more specifically, its venom sac extract (VSE) possesses deoxyribonuclease activity that exerts an effect both on insects as well as on mammals. We have now examined the effect of hornet VSE on primary culture of rat cortical neurons. Judging on the basis of our results, VSE induces a rapid cell death by a) permeabilizing the cell membrane, b) inducing DNA breaks, and c) cleaving the nuclear protein poly-ADP-ribose polymerase (PARP-1), thereby preventing DNA repair.     

Thematic Review Series: Bile Acids: Bile acids as regulatory molecules

In the past, bile acids were considered to be just detergent molecules derived from cholesterol in the liver. They were known to be important for the solubilization of cholesterol in the gallbladder and for stimulating the absorption of cholesterol, fat-soluble vitamins, and lipids from the intestines. However, during the last two decades, it has been discovered that bile acids are regulatory molecules. Bile acids have been discovered to activate specific nuclear receptors (farnesoid X receptor, preganane X receptor, and vitamin D receptor), G protein coupled receptor TGR5 (TGR5), and cell signaling pathways (c-jun N-terminal kinase 1/2, AKT, and ERK 1/2) in cells in the liver and gastrointestinal tract. Activation of nuclear receptors and cell signaling pathways alter the expression of numerous genes encoding enzyme/proteins involved in the regulation of bile acid, glucose, fatty acid, lipoprotein synthesis, metabolism, transport, and energy metabolism. They also play a role in the regulation of serum triglyceride levels in humans and rodents. Bile acids appear to function as nutrient signaling molecules primarily during the feed/fast cycle as there is a flux of these molecules returning from the intestines to the liver following a meal. In this review, we will summarize the current knowledge of how bile acids regulate hepatic lipid and glucose metabolism through the activation of specific nuclear receptors and cell signaling pathways.     

Increased serum bile acids as a possible biomarker of hepatotoxicity in Brazilian workers exposed to solvents in car repainting shops

Abstract

The objective was to evaluate total serum bile acids (SBA) as a biological marker of hepatotoxicity in car painters exposed to organic solvents and to compare their performance with classic biochemical parameters of liver function. SBA were analysed in a selected group of workers (n=57) occupationally exposed to a mixture of organic solvents and in a control group (n=51). In addition, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TB), gamma-glutamyltransferase (GGT) and alkaline phosphatase (ALP) were determined in the two groups. Urinary hippuric acid was measured in all samples. Statistical analysis of the data revealed a significant increase in the concentration of SBA, AST, ALP and TB in exposed workers compared with controls (Mann–Whitney, p≤0.05). However, SBA was the parameter most frequently altered in exposed workers and showed higher significance between the two groups (chi-square test) compared with the upper limit of the reference range (8?µmol?l^?1). In conclusion, SBA can be considered to be a sensitive parameter of hepatotoxicity induced by organic solvents than the traditional tests and it can be used as an biological marker of subclinical liver injury.     

Increased serum bile acids as a possible biomarker of hepatotoxicity in Brazilian workers exposed to solvents in car repainting shops.

The objective was to evaluate total serum bile acids (SBA) as a biological marker of hepatotoxicity in car painters exposed to organic solvents and to compare their performance with classic biochemical parameters of liver function. SBA were analysed in a selected group of workers (n=57) occupationally exposed to a mixture of organic solvents and in a control group (n=51). In addition, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TB), gamma-glutamyltransferase (GGT) and alkaline phosphatase (ALP) were determined in the two groups. Urinary hippuric acid was measured in all samples. Statistical analysis of the data revealed a significant increase in the concentration of SBA, AST, ALP and TB in exposed workers compared with controls (Mann-Whitney, p相似文献   

Bile acids. XXXVII. Identification of the 3 beta isomers of allocholic and allochenodeoxycholic acids as metabolites of 5 -cholestanol in the rat

Bile was collected for 18-24 days from adult male rats with cannulated bile ducts that had received intraperitoneally 0.8 mg of 5alpha-[4-(14)C, 3alpha-(3)H]cholestan-3beta-ol. Bile from the first 2 days containing 14.2% of the administered (14)C and 3.3% of the (3)H was hydrolyzed, and the bile acids were separated by acetic acid partition chromatography. The previously unidentified metabolite more polar than cholic and allocholic acids was identified by isotopic dilution as 3beta,7alpha,12alpha-trihydroxy-5alpha-cholanic acid and represented 3% of the biliary (14)C and 15% of the (3)H. Similarly, 3beta,7alpha-dihydroxy-5alpha-cholanic acid was identified in fractions more polar than allochenodeoxycholic acid and represented 0.6% of the biliary (14)C and 8% of the (3)H. More polar fractions contained 4% of the (14)C and 31% of the (3)H in unidentified metabolites.     

Bile acids in the fetal rat: Effect of maternal bile duct ligation

The effect of bile duct ligation during pregnancy in rats (thereby increasing maternal plasma bile acids levels) on the bile acid content and composition in the fetus was examined. In spite of 30-fold increase in maternal plasma cholic acid, the bile acid content in the fetus of bile duct ligated rats was significantly lower (P <0.05) with a significant reduction in cholic acid content. Plasma cholesterol levels of fetuses from bile duct ligated rats were also significantly lower (p <0.05). In addition to the commonly expected bile acids, gas-liquid Chromatographic analysis of the fetal bile acid pool showed peaks corresponding to several secondary bile acids. These results suggest that the transfer of primary bile acids of maternal origin into the fetus is minimal.     

Bile acids. 38. Conversion of 5 -cholestane-3 ,7 -diol to allo bile acids by the rat

5alpha-[4-(14)C, 3alpha-(3)H]Cholestane-3beta,7alpha-diol was prepared from individual samples of 5alpha-[3alpha-(3)H]cholestane-3beta,7alpha-diol and 5alpha-[4-(14)C]cholestane-3beta,7alpha-diol, each derived from 3beta-acetoxycholest-5-en-7-one. Bile was collected for 11 days from adult male rats, with cannulated bile ducts, that had received intraperitoneally 0.90-0.92 mg of the doubly labeled diol. Bile from the first 10 hr, containing 63% of the administered (14)C and 6% of the (3)H, was hydrolyzed, and the bile acids were separated by acetic acid partition chromatography. Allochenodeoxycholic and allocholic acids contained at least 20.6% and 48.6%, respectively, of the (14)C retained in the biliary acids. Small amounts of (14)C (2.5% and 1.9%, respectively) were present in the 3beta isomers of these acids, but the tritium content totaled more than half of that found in the bile acid fraction. No evidence was obtained for presence of the extensive quantities of the allomuricholates.     

Hornetin: the lethal protein of the hornet (Vespa flavitarsus) venom

By gel permeation on a Fractogel TSK HW 50 column followed by ion-exchange chromatography on carboxymethylcellulose CM 52, a lethal protein, designated hornetin, was purified from the venom of Vespa flavitarsus. Hornetin is a highly basic protein (pI 10.2) with a molecular mass of about 32 kDa. Its amino acid composition is characterized by a high content of lysine, aspartic and glutamic acid, and is devoid of tryptophan and cysteine. The lack of cysteine in the molecule is distinct from other known vespid venom proteins of comparable size. The i.v. LD50 of the toxin is 0.42 microgram per g mouse. Assayed on the red blood cells of the mouse and guinea-pig as well as isolated nerve muscle preparations of the chick and mouse, hornetin showed direct hemolytic activity and presynaptic neurotoxicity at microgram level and displayed musculotropic effect at higher concentrations.     

Bile acids in the rat: studies in experimental occlusion of the bile duct

Bile acids in the plasma, urine, and small intestine of adult male rats with occluded bile ducts have been studied using a method of high specificity for their determination. After bile duct ligation cholic acid rapidly accumulates in the plasma for 8 hr, remains high for a further 8 hr, and subsequently diminishes; bile acids disappear from the small intestine. During the first 12 hr after bile duct ligation the excretion of trihydroxy acids in the urine was 10 times that of the dihydroxy acids. Subsequently the two excretion rates became equal. Because bile acids have been implicated in the etiology of hepatic damage following bile duct ligation, studies have been made of the effect on the liver of removing (with cholestyramine) and supplementing (with cholic acid) the intestinal bile acid pool. The addition of cholestyramine to the stock diet prevented the rise in trihydroxy bile acids after bile duct ligation, but did not prevent the development of histological abnormalities in the liver. Supplementing the diet with cholic acid raised the plasma cholic acid levels but had little effect on the hepatic histological findings.     

Orientotoxin--a new presynaptic neurotoxin from the venom of the giant hornet Vespa orientalis

Orientotoxin, a novel presynaptically acting neurotoxin from the venom of giant hornet Vespa orientalis, has been isolated by gel filtration and ion exchange chromatography and characterized. The toxin has a molecular mass of 18,000. Highly purified preparations of orientotoxin possessed clearly manifested lysophospholipase activity and can block both induced and spontaneous release of neurotransmitter from the presynaptic nerve membrane.