The effects of estrogen on the α2-adrenergic receptor subtypes in rat uterine function in late pregnancy in vitro

Aim To assess the effect of 17β-estradiol pretreatment on the function and expression of α2- adrenergic receptors (ARs) subtypes in late pregnancy in rats. Methods Sprague-Dawley rats (n = 37) were treated with 17β-estradiol for 4 days starting from the 18th day of pregnancy. The myometrial expression of the α2-AR subtypes was determined by real time polymerase chain reaction and Western blot analysis. In vitro contractions were stimulated with (-)-noradrenaline, and its effect was modified with the selective antagonists BRL 44408 (α2A), ARC 239 (α2B/C), and spiroxatrine (α2A). The cyclic adenosine monophosphate (cAMP) accumulation was also measured. The activated G-protein level was investigated by guanosine 5’-O-[gamma-thio]triphosphate (GTPγS) binding assay. Results 17β-estradiol pretreatment decreased the contractile effect of (-)-noradrenaline via the α2-ARs, and abolished the contractile effect via the α2B-ARs. All the α2-AR subtypes’ mRNA was significantly decreased. 17β-estradiol pretreatment significantly increased the myometrial cAMP level in the presence of BRL 44408 (P = 0.001), ARC 239 (P = 0.007), and spiroxatrine (P = 0.045), but did not modify it in the presence of spiroxatrine + BRL 44408 combination (P = 0.073). It also inhibited the G-protein-activating effect of (-)-noradrenaline by 25% in the presence of BRL 44408 + spiroxatrine combination. Conclusions The expression of the α2-AR subtypes is sensitive to 17β-estradiol, which decreases the contractile response of (-)-noradrenaline via the α2B-AR subtype, and might cause changes in G-protein signaling pathway. Estrogen dysregulation may be responsible for preterm labor or uterine inertia via the α2-AR

Thermotropic and structural effects of poly(malic acid) on fully hydrated multilamellar DPPC–water systems

The thermotropic and structural effects of low molecular weight poly(malic acid) (PMLA) on fully hydrated multilamellar dipalmitoylphosphatidylcholine (DPPC)-water systems were investigated using differential scanning calorimetry (DSC), small-angle X-ray scattering (SAXS), and freeze-fracture transmission electron microscopy (FFTEM). Systems of 20 wt% DPPC concentration and 1 and 5 wt% PMLA to lipid ratios were studied. The PMLA derivatives changed the thermal behavior of DPPC significantly and caused a drastic loss in correlation between lamellae in the three characteristic thermotropic states (i.e., in the gel, rippled gel and liquid crystalline phases). In the presence of PBS or NaCl, the perturbation was more moderate. The structural behavior on the atomic level was revealed by FTIR spectroscopy. The molecular interactions between DPPC and PMLA were simulated via modeling its measured infrared spectra, and their peculiar spectral features were interpreted. Through this interpretation, the poly(malic acid) is inferred to attach to the headgroups of the phospholipids through hydrogen bonds between the free hydroxil groups of PMLA and the phosphodiester groups of DPPC

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In vitro Interaction between Fumonisin B1 and the Intestinal Microflora of Pigs

The caecal chyme of pigs was incubated anaerobically in McDougall buffer with and without fumonisin B1 (5 &mu;g/ml) for 0, 24 and 48 h. The plate count agar technique was applied for enumerating the amount of bacteria including aerobic, anaerobic bacteria, coliform, Escherichia coli and Lactobacillus sp. The quantitative polymerase chain reaction was also performed to estimate the number of copies of the total bacteria, Lactobacillus, Bacteroides and Prevotella. No significant differences in the amount of bacterial groups between the experimental (buffer, chyme, and fumonisin B1) and control 1 groups (buffer + chyme) were observed in both methods. Fumonisin B1 and hydrolysed fumonisin B1 concentration were analysed by liquid chromatograghy &ndash; mass spectrometry. There was no significant difference in FB1 concentration between the experimental and the control 2 group (buffer and fumonisin B1) at 0 h incubation, 5.185 &plusmn; 0.174 &mu;g/ml com&shy;pared with 6.433 &plusmn; 0.076 &mu;g/ml. Fumonisin B1 concentration in the experimental group was reduced to 4.080 &plusmn; 0.065 &mu;g/ml at 24 h and to 2.747 &plusmn; 0.548 &mu;g/ml at 48 h incubation and was significantly less than that of in the control group. Hydrolysed fumonisin B1 was detected after 24 h incubation (0.012 &plusmn; 0 &mu;g/ml). At 48 h incubation time, hydrolysed fumonisin B1 concentration was doubled to 0.024 &plusmn; 0.004 &mu;g/ml. These results indicate that fumonisin B1 can be metabolised by caecal microbiota in pigs though the number of studied bacteria did not change

Fumonisin B1 exposure increases Hsp70 expression in the lung and kidney of rats without inducing significant oxidative stress

The objective of this experiment was to determine whether fumonisin B1 (FB1) added to the diet of rats in a dose of 50 mg/kg changes the production of heat shock protein 70 (Hsp70) in the lungs and kidney of rats. We also studied the effect of this mycotoxin on the antioxidant system of the body. Mature (8 weeks old) male Wistar Crl:WI BR rats (n = 6/group) were fed the toxin-containing diet for 5 days. FB1 resulted in a 7% body weight reduction without significantly changing the feed intake. Western blot analysis of the lungs and kidney demonstrated a substantial (1.4-fold and 1.8-fold, respectively) increase in Hsp70 expression. Alterations could not be detected in the clinical chemical parameters (total protein, albumin, total cholesterol, glucose, creatinine and urea concentrations, and aspartate aminotransferase activity). There was no statistically significant change in malondialdehyde concentrations and the measured antioxidant parameters (the amount of reduced glutathione, GSH and glutathione peroxidase activity, GPx) in the blood plasma, lung and kidney tissue. Thus, it can be concluded that FB1 did not induce oxidative stress in the lungs and kidney, but increased Hsp70 production

Individual and combined effects of feed artificially contaminated with with fumonisin B 1 and T-2 toxin in weaned rabbits

Co-contamination of feed and feed raw materials with two or more mycotoxins is frequently reported, however, only a few studies have investigated the combined effects of low doses of multiple mycotoxins. In the present study the individual and combined effects of 10 mg/kg fumonisin B1 and 2 mg/kg T-2 toxin (n=12/group) were investigated in weaned rabbits. Mycotoxin contaminated feed was produced by adding fungal cultures of Fusarium verticillioides and Fusarium sporotrichioides, and fed to 40 days old rabbits during 28 days. Feed intake and body weight were measured weekly, serum biochemistry and antioxidant parameters on day 0, 14 and 28, while histopathological examination and comet assay were performed at the end of the experiment. T-2 exposure both alone and in combination resulted in 15-18% less final body weight compared to the control and FB1 treatment. There was a significant increase in the concentration of plasma total protein, albumin, fructosamine and creatinine in the group treated with FB1 compared to the control. The liver and the kidney of most animals treated with T-2 toxin, FB1 and their combination showed pathological changes, occurring more frequent in animals exposed to both toxins. T-2 resulted in depletion of lymphocytes in the spleen. FB1 and T-2 exerted synergistic effect on the antioxidant/oxidative parameters after 2 weeks of exposure, manifesting in less glutathione and glutathione peroxidase, while more malondialdehyde was produced. Both toxins caused DNA damage in the lymphocytes, which was more pronounced in the group fed T-2 toxin and T-2 combined with FB1, without additive or synergistic effects