The effect of exogenous oxytocin on streptozotocin (STZ)-induced diabetic adult rat testes.

Oxytocin (OXY) plays a crucial role in reproduction. The aim of this study is to investigate the therapeutic and protective effects of oxytocin treatment on streptozotocin (STZ) induced diabetes in testicular tissue. The rats were randomly divided into four experimental groups: (I) Control Group, (II) STZ induced Diabetic Group (STZ Group), (III) STZ induced Diabetic Group with Pre-Oxytocin treatment (Pre-OXY Group) and (IV) STZ induced Diabetic Group with Post-Oxytocin treatment (Post-OXY Group); each group contains six animals. The rats whose blood glucose levels were more than 200mg/dl were included to the experiment. At the end of the 4th week, testes tissue samples were taken to be processed for light microscopy and transmission electron microscopy. Malondialdehyde (MDA), Glutathione (GSH) and Advanced Oxidation Protein Products (AOPP) levels were determined biochemically in blood samples. Testicular tissue samples stained with Hematoxylin and Eosin (H&E) and Periodic acid-Schiff (PAS) reaction were evaluated under light microscope. The histopathological damage score of testicular tissue, which was significantly increased in STZ group, was decreased by oxytocin treatment. According to biochemical data, MDA and AOPP levels have been increased in the blood of STZ Group compared to the Control Group whereas they decreased significantly in Oxytocin-treated Groups compared to STZ Group. GSH levels were significantly decreased in the blood of STZ Group and increased in the blood of Oxytocin-treated Groups compared to STZ Group. In conclusion, oxytocin has a potential protective effect on the testes tissue of STZ-induced diabetic rats.

Plasma levels of growth arrest specific protein 6 are increased in idiopathic recurrent pregnancy loss.

AIM: The aim of this study was to determine plasma Growth Arrest Specific Protein 6 (GAS6) protein levels in patients with unexplained recurrent pregnancy loss (RPL) and compare them to those of pregnant and to healthy non-pregnant women with a history of at least one live delivery. PATIENTS AND METHODS: A total of 205 women were included in the study. Of these, 68 were diagnosed with unexplained RPL and were not pregnant at the time of the study. The second group consisted of 67 pregnant women in the third trimester of pregnancy. The third group constituted the control group of 70 healthy non-pregnant women who had at least one live birth. Plasma levels of GAS6 protein were measured by ELISA. RESULTS: Mean plasma GAS6 levels were found to be different between RPL group and healthy non-pregnant women (12.17 ± 4.39 µg/L  and 9.18 ± 4.65, respectively, p = 0.0013). Although it was not statistically significant, plasma levels of GAS6 in the third trimester pregnant group (10.65 ± 3.74 µg/L) were found to be slightly higher than non-pregnant controls, and slightly lower than RPL group. In comparison of those with 2 or more than 2 pregnancy losses in the RPL group, there was no difference between these two subgroups in terms of GAS6 levels (12.39 ± 4.27 µg/L and 11.83 ± 4.63 µg/L, respectively, p = 0.62). CONCLUSIONS: Our findings revealed that plasma GAS6 levels were increased in patients with RPL. The prothombotic and proinflammatory nature of the GAS 6 protein makes it a likely culprit involved in the pathologic process in patients with RPL. Further studies are warranted to determine the possible role of GAS6 protein in the pathophysiology of idiopathic RPL.

Apo A-I binding to platelets detected by flow cytometry.

Lipoprotein-platelet interactions are very important in atherosclerosis and thrombosis. Several studies have been carried out on specific binding of various lipoproteins to platelets. But there is considerable disagreement about the details of these binding sites. Although low-density lipoprotein (LDL) receptors of several cells have been studied extensively, there is little datum about high-density lipoprotein (HDL) receptors. Apolipoprotein (apo) A-I may play a major role in the determination of the specificity of HDL receptors. In this study, binding of apo A-I to platelets was investigated by using a flow cytometric method. Citrated blood samples were obtained from five healthy and seven hypercholesterolemic subjects. Apo A-I antibody was incubated with the citrated whole blood before and after activation with ADP or thrombin receptor agonist peptide (TRAP). Then fluorescein isothiocyanate (FITC)-labeled secondary antibodies were added and analyzed on a Becton-Dickinson FACSort flow cytometer. In the hypercholesterolemic group, apo A-I binding to platelets was found to be significantly decreased after activation with TRAP (P<.05), but not after activation with ADP. In the control group, after platelet activation with ADP or TRAP, the apo A-I MFI values were not found to be significantly different from the values of resting platelets (P>.05). In this study, we demonstrated that apo A-I can bind to platelets, and this supports the hypothesis that apo A-I may play a major role in HDL binding to platelets.

Avidin-biotin ELISA for measurement of prothrombin in human plasma.

In this study, IgG was partially purified from rabbit antisera against human normal prothrombin. The presence of antibodies against prothrombin was shown by Ouchterlony's double immunodiffusion method. This IgG was adsorbed to microwells and an enzyme-linked immunosorbent assay (ELISA) method was developed by using biotinylated prothrombin. We used alkaline phosphatase as the labeling enzyme. Plasma prothrombin concentrations in 30 healthy individuals and 15 patients with liver cirrhosis were measured using this ELISA method. Prothrombin levels of the cirrhotic patients (61+/-36 microg/ml) were significantly lower than those of the control values (117+/-43 microg/ml) (P<.001). In addition to the ELISA method in the same samples, we assayed prothrombin levels by nephelometry and prothrombin activities using staphylocoagulase and chromogenic substrate. With nephelometry, in healthy individuals, a mean value of 104+/-17 microg/ml and, in cirrhotic patients, a mean value of 51+/-26 microg/ml of prothrombin levels were detected. The prothrombin activities were 10532+/-2429 and 2433+/-330 U/l, respectively. In both methods, the prothrombin values of cirrhotic patients were also significantly lower than those of the healthy individuals (P<.001). The correlation coefficient between ELISA and nephelometry was r=.90, P<.01, and between ELISA and the amidolytic assay was r=.69, P<.01. Prothrombin times (PT) of cirrhotic patients (18.5+/-2.3 s) were significantly longer and albumin (2.4+/-0.5 g/dl) levels were significantly lower than those of normal values (P<.001). PT were negatively correlated with prothrombin levels assayed by ELISA (r=-.59, P<.01). This enzyme immunoassay technique can be used to measure prothrombin levels in plasma.

Determination of the N-terminal amino acid sequence of the purified prothrombin from a patient with liver cirrhosis.

The reasons for the decreased functional activity of prothrombin in liver diseases are still speculative. When a highly purified preparation of prothrombin from a patient with liver cirrhosis is available, the cause of prothrombin abnormalities may be researched on a molecular basis. In this study, prothrombin (6.7 mg) was purified from the ascites fluid (1130 mL) of a patient with liver cirrhosis by barium citrate adsorption, ammonium sulfate elution, DEAE Sephacel and Heparin Sepharose CL-6B column chromatography steps. The molecular weight of this prothrombin was the same as that of normal prothrombin purified from a normal plasma pool. The specific activities were found to be 3.36 U/mg in the one stage clotting assay and 28.9 U/mg in the staphylocoagulase/chromogenic substrate assay, while the normal prothrombin specific activities were 3.92 U/mg and 30.1 U/mg respectively. When N-terminal amino acid sequence analysis was carried out, it was seen that the first 20 residues were identical to the normal human prothrombin excepting the Gla at position #14.

Nephrotoxicity of gentamicin and co-trimoxazole combination in rats.

1. The nephrotoxicity of gentamicin is well known. However, little information is available regarding the combined effects of gentamicin plus co-trimoxazole (sulfamethoxazole-trimethoprim). Therefore, Wistar rats were treated daily with 100 mg/kg gentamicin or 100 mg/kg gentamicin plus 30 mg/kg trimethoprim-150 mg/kg sulfamethoxazole for 14 days. 2. Serum biochemical parameters were measured on days 0, 8 and 15, and histopathological examinations of kidneys were performed on day 15, one day following end of treatment. Gentamicin treated rats exhibited a 63% increase in blood urea nitrogen (BUN), a 124% increase in uric acid, and a 63% decrease in serum potassium levels on day 15. 3. The combination of gentamicin plus co-trimoxazole partially ameliorated these effects. With the three drug combination no change occurred in BUN, and only a 30% decrease occurred in serum potassium levels. 4. While serum creatinine levels significantly increased following gentamicin, the co-administration of co-trimoxazole resulted in a significant decrease (30%) in creatinine. Histopathological examinations of kidneys suggested a lower degree of nephrotoxicity in rats treated with gentamicin plus co-trimoxazole as compared to animals treated with gentamicin alone. 5. The results support the importance of monitoring serum biochemical parameters when treating with gentamicin or gentamicin plus co-trimoxazole.