Tannic acid exhibits anti-inflammatory effects on formalin-induced paw edema model of inflammation in rats.

The aim of the study was to determine the relationship between anti-inflammatory effects of the natural polyphenolic compound tannic acid (CAS number: 1401-55-4) and myeloperoxidase (MPO) enzyme activity in paw edema model. Thirty-five female rats were divided into five groups. The paws of rats were injected subcutaneously in the plantar surface with formalin except for the control group. Indomethacin and tannic acid were intraperitoneally administered 1 h after formalin injection. The paws volume was measured by using vernier caliper. MPO enzyme activity was determined using 4-aminoantipyrine-phenol solution as the substrate for MPO-mediated oxidation by H2O2. About 17% and 13% edema inhibition has detected in the indomethacin-applied group, at the measurements run every other hour right after the treatment. An inhibition of 16% was found at the group treated with 25 mg/kg tannic acid. However, in the group treated with 50 mg/kg tannic acid, 15% and 7% of the edema inhibition was observed. Serum and paw tissue MPO activities were decreased in treated groups with indomethacin and tannic acid according to formalin control group. Our study results suggest that tannic acid may contribute to the treatment of inflammation by decreasing MPO enzyme activity, but the molecular mechanism is still not clear.

Effects of genetic variations in the genes encoding NOD1 and NOD2 on type 2 diabetes mellitus and insulin resistance.

WHAT IS KNOWN AND OBJECTIVE: Nucleotide-binding oligomerization domain (NOD) 1 and NOD 2 are members of the NOD-like receptor (NLR) family and contain a caspase recruitment domain. NLRs are located in the cytosol, bind bacterial and viral ligands and play a key role in the realization of innate and adaptive immune response, inflammation, apoptosis and reactive oxygen species generation. Insulin resistance (IR) is a leading cause of type 2 diabetes mellitus (T2DM) and associated with obesity, inflammation and pro-inflammatory responses. NOD1 and NOD2 gene variants may affect the risk of chronic inflammation, insulin resistance and T2DM by shifting the balance between pro- and anti-inflammatory cytokines. The aim of our study was to determine whether the NOD1/2 gene variants might contribute to the risk of T2DM and IR. METHODS: The rs5743336 variant of NOD1 and rs2066847 variant of NOD2 were analysed by PCR-RFLP analysis in 200 subjects (T2DM: n = 100; healthy controls: n = 100) of Turkish origin. PCR products were digested with the AvaI and ApaI restriction enzymes. For the NOD1 site, the presence of the G allele was indicated by cleavage of the 379 bp amplified PCR product that yielded 209-bp and 170-bp fragments. For the NOD2 site, 151-bp PCR products were cleaved and yielded 130-bp and 21-bp fragments when the WT-insC mutation was present. Comparisons of the genotypes between controls and patients were performed by chi-square tests. RESULTS AND DISCUSSION: The genotypes of the rs5743336 variant of NOD1 and the rs2066847 variant of NOD2 are presented, and no significant differences were observed in the genotype frequencies of the NOD1 and NOD2 variants between the healthy controls and T2DM patients (P > 0·05). According to our preliminary data, NOD1/2 gene variants are not linked with T2DM and IR. WHAT IS NEW AND CONCLUSION: This study is the first to look for possible association of the genotype frequencies of NOD1 and NOD2 genes with T2DM and IR. The significant finding of this report is that the rs5743336 and rs2066847 variations in the NOD1/2 gene are not associated with T2DM and IR risk in patients of Turkish origin.

A study of the possible association of plasminogen activator inhibitor type 1 4G/5G insertion/deletion polymorphism with susceptibility to schizophrenia and in its subtypes.

WHAT IS KNOWN AND OBJECTIVE: Inhibition of the fibrinolytic system may occur at the level of plasminogen activation, mainly by PAI-1. Mental and physical stress caused to alterations of platelet function, and also decreased to fibrinolytic activity. Furthermore, stress-induced thrombosis regulation was proposed to be by PAI-1 in schizophrenia patients. In this study, the distribution of genotypes and frequency of alleles of the plasminogen activator inhibitor type 1 (PAI-1) gene 4G/5G polymorphism in different Turkish clinical schizophrenia subtypes was investigated for its role in schizophrenia development. METHODS: The clinical schizophrenia subtypes include paranoid, catatonic, disorganized, undifferentiated and residual, as diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition IV (DSM-IV). Samples of genomic DNA (250 total, including 150 schizophrenia patients and 100 healthy subjects) were analysed. PAI-1 4G/5G genotyping was performed by polymerase chain reaction-allele-specific amplification. PCR products were separated by 2% agarose gel electrophoresis and then visualized. RESULTS AND DISCUSSION: The genotype distributions (P = 0·136) and allele frequencies (P = 0·721 for 4G, P = 0. 097 for 5G) were not significantly different between patients with schizophrenia and control subjects for the 4G/5G polymorphism. Similar results were also found for the genotype distributions (P = 0·640) and allele frequencies (P = 0·763 for 4G, P = 0·448 for 5G) in the clinical schizophrenia subtypes compared to the each other. WHAT IS NEW AND CONCLUSION: We conclude that PAI-1 4G/5G polymorphism was not significantly associated with schizophrenia or its subtypes in the Turkish population. However, we recognize that with our sample sizes, we cannot exclude weak associations.

Association of paraoxonase 1 (PON1) gene polymorphisms and concentration with essential hypertension.

Human serum paraoxonase 1 (PON1) is carried by high-density lipoprotein in blood circulation and is shown to be effective in preventing oxidized phospholipids carried by low-density lipoprotein particles, thus it acts as an antioxidant. Polymorphism in this gene has been investigated for many metabolic diseases, but it is not thought to be a genetic risk factor for essential hypertension. The aim of this study was to determine whether there was an association between PON1 gene polymorphisms and concentration with essential hypertension. The study population was comprised of 100 patients with essential hypertension and 100 healthy controls. One promoter region [C(-108)T] and two coding region (Q192R and L55M) polymorphisms in the PON1 gene were genotyped in individuals by using the TaqMan assay. Plasma PON1 concentration in all volunteers was also measured spectrophotometrically by the enzyme-linked immunosorbent assay method. The genotype and allele frequencies of the PON1 C(-108)T polymorphism showed significant differences between the essential hypertensive and control groups (CT vs. CC: p<0.001; T allele vs. C allele: p<0.001). There was no significant difference for the PON1 L55M polymorphism between the groups, while the heterozygote genotype of the PON1 Q192R polymorphism showed significant difference (p = 0.03). The PON1 concentration was also found to be significantly lower in hypertensive patients (p < 0.001). Decline in the level of PON1 gene may be one of the main factors in the development of essential hypertension, and the PON1 C(-108)T polymorphism may have a prognostic value in the patients with essential hypertension.

Effect of retinyl acetate on transglutaminase 2 activity in carcinogen treated rat liver.

Transglutaminase 2 (TG2) has been implicated in wound healing, cellular differentiation, apoptosis and cell survival. TG2 activity increases following acute and chronic liver injury; however, the role of TG2 in tumors, is controversial. TG2 is a retinoid-inducible enzyme. We investigated the effects of retinyl acetate (RA) on the activity and levels of TG2 during the initiation and promotion stages of liver cancer. p-Dimethylaminoazobenzene (p-DAB) was used as initiator and 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) was used as promoter in our model of carcinogenesis. Rats were divided into four groups of 24: control, corn oil control, p-DAB + TCDD, and p-DAB + TCDD + RA. Six rats from each group were sacrificed at days 30, 60, 90 and 120. TG2 activity decreased in the p-DAB + TCDD treated group, but TG2 immunostaining scores did not change by days 90 and 120. Neither TG2 enzyme activity nor the immunostaining score of TG2 protein changed in the tissues of the p-DAB + TCDD + RA group by days 90 and 120. TG2 activity was not be ameliorated by RA during the initiation or promotion stages of carcinogen induced liver cancer.

Acute stroke in relation to homocysteine and methylenetetrahydrofolate reductase gene polymorphisms.

AIM: Some methylenetetrahydrofolate reductase (MTHFR) gene mutations cause hyperhomocysteinemia and homocystinuria. These may be important risk factors for cardio and cerebrovascular diseases. We investigated whether the MTHFR C677T and A1298C polymorphisms contribute to hyperhomocysteinemia and increase the risk factor for stroke. METHODS: A total of 203 acute stroke patients and 55 controls were recruited. Polymorphisms were determined by using polymerase chain reaction-restriction fragment length polymorphism (RFLP) and plasma total homocysteine levels were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS AND CONCLUSIONS: There were no significant differences between C677T and A1298C genotypes and allele frequencies in the stroke patients and controls. Total plasma homocysteine level was higher in the 677TT and 1298AA genotypes in stroke patients and especially small-vessel disease patient subgroup. Age, number of males, systolic-diastolic blood pressures, creatinine, vitamin B(12) and homocysteine levels were significantly high among stroke patients. Age, sex, systolic blood pressure and HDL-C were determined as risk factors for homocysteine levels. We also determined that the effect of A1298C polymorphism on homocysteine was not as high as that of C677T polymorphism in acute stroke patients. We conclude that the MTHFR genotype may be a modest risk factor for stroke in Turkish population.

Frequency of angiotensin-converting enzyme gene polymorphism in Turkish type 2 diabetic patients.

We aimed to investigate the angiotensin-converting enzyme (ACE) gene polymorphism, ACE activity and their associations with diabetic complications in Turkish patients with type 2 diabetes mellitus. A total of 143 patients and 133 controls were screened for ACE gene I/D polymorphism by using polymerase chain reaction. Serum ACE activities were determined spectrophotometrically. There was no significant difference in the distribution of ACE I/D genotypes between patients and controls. The patients with DD genotype had a higher ACE activity than those with ID and II. Hypertensive diabetic patients with DD genotype had higher ACE activities than those with ID and II. There was no significant difference in the distribution of ACE I/D genotypes between patients with and without nephropathy, retinopathy and hypertension except for patients with and without neuropathy. In patients with DD genotype, creatinine clearance correlated with duration of diabetes. The grade of retinopathy was correlated with duration of diabetes in DD and ID genotypes. The highest ACE activity was measured in hypertensive diabetics with DD genotype. ID genotype was suggested to be a risk factor and II was suggested to be protective for diabetic neuropathy. The DD and ID genotypes might be a predictor for the development of retinopathy in relation to duration of diabetes.

Frequency of angiotensin-converting enzyme gene polymorphism in Turkish hypertensive patients.

Hypertension is a multifactorial disease, in which genetic factors play an important role. This study was carried out to determine angiotensin-converting enzyme levels and angiotensin-converting enzyme gene polymorphism in Turkish hypertensive patients, and to establish whether there is an association of angiotensin-converting enzyme gene polymorphism with clinical and echocardiographic parameters. We have investigated the association among the allelic distribution of the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme gene identified by polymerase chain reaction, angiotensin-converting enzyme activity determined spectrophotometrically, cardiac morphology and function assessed by means of echocardiography. Distribution of angiotensin-converting enzyme gene I/D polymorphism and allele frequencies in hypertensive patients was not significantly different from controls. D allele frequency was 51.7% in hypertensives vs. 51.9% in controls and I allele 48.3 vs. 48.1%, respectively. The level of angiotensin-converting enzyme activity was significantly higher in the patients homozygotes for D allele (DD = 59.93 U/l) than in heterozygotes (ID = 39.49) and in homozygotes for I allele (II = 40.28 U/l). In addition to these, the level of angiotensin-converting enzyme activity was significantly lower in the ID and especially II patients receiving ACE inhibitors than the others. Also, it was determined that left atrium diameter was larger in the patients homozygotes for I allele than the others.

The effects of acarbose and Rumex patientia on liver ultrastructure in streptozotocin-induced diabetic (type II) rats.

The aim of this study was to investigate the effects of acarbose and Rumex patientia on liver ultrastructure in streptozotocin (STZ)-induced diabetic (type II) rats. Forty-two-day-old, neonatal Wistar albino rats were used. They were divided into six groups. STZ was injected into groups 4, 5 and 6 on postnatal day 2. Groups 1 and 5 received water, groups 2 and 6 received 2% decoction of R. patientia grain and groups 3 and 4 received 40 mg acarbose/100 g feed. During the experimental period, blood glucose levels were checked periodically and HbA1c levels were measured from cardiac blood at the end of the experiment. In addition, liver tissue was examined by electron microscopy. Our results showed that glucose and HbA1c levels, which are increased by STZ, were decreased by acarbose and R. patientia. In group 5, most of the mitochondria of hepatocytes were swollen and some hepatocytes contained lipid granules in their cytoplasm. In group 4, no pathological changes were observed in hepatocytes, but some lysosomes were found in their cytoplasms. In group 6, mitochondrial changes were minimal compared with those in group 5, and no lipid granules were observed in hepatocytes.

The effect of alpha-tocopherol and pentoxyfilline on ischemia-reperfusion induced liver injury in rats.

BACKGROUND/AIM: In the present study our purpose was to investigate the effect of pentoxyfilline, that plays a role in microcirculation and tissue oxygenation, alone and in combination with an antioxidant vitamin E on tissue damage in the rat liver induced by ischemia-reperfusion. METHODOLOGY: Thirty-one albino rats were divided into four groups. Rats in group I (n= 7), group II (n= 8) and group III (n= 8) were given, respectively, pentoxyfilline (25 mg/kg), pentoxyfilline and vitamin E in combination (25 mg/kg and 50 mg/kg, respectively) and equal volume of saline solution intraperitoneally for 7 days. Rats in group IV (n= 8) served as controls and received no treatment. On day 7 ischemia was induced by cross-clamping the hepatic artery, portal vein and left branch of the biliary duct for 30 minutes. Malondialdehyde (MDA) and catalase activity were assessed in tissue sample, and the level of ALT was measured in serum obtained after reperfusion for 30 minutes. Histological examination of tissue sample was also carried out. RESULTS: There was no significant difference in ALT level between three study groups. Group I and group II had significant lower MDA and catalase levels than those of group III. The results of histopathologic examination in group I and group II were better than that of group III. CONCLUSION: Our findings suggested that the treatment of pentoxyfilline alone and in combination with vitamin E decreased liver damage induced by ischemia-reperfusion and that the effect of latter was more effective but the difference between the two treatment patterns was not statistically significant.

Effects of S-CMC on the cisplatin toxicity in rats.

In the present study, some toxic effects of cisplatin are evaluated in rats. It was also investigated whether S-carboxymethylcysteine (S-CMC), a free radical scavenger, protects the experimental animals from the toxic effects of cisplatin. The 1st, 2nd, 4th and 5th groups received physiological saline, dimethyl sulphoxide, and S-CMC (100 and 500 mg/kg i.p.) for 3 days, respectively. The 3rd group received cisplatin (5 mg/kg i.p.) 12 h before sacrifice. The 6th and 7th groups received S-CMC (100 and 500 mg/kg i.p., respectively); additionally, these groups received cisplatin (5 mg/kg i.p.) 12 h before the rats were sacrificed. 5 mg/kg cisplatin decreased significantly serum creatinine and glutamic-oxaloacetic and glutamic-pyruvic transaminase levels as well as leucocyte counts. Although S-CMC did not change the effects of cisplatin on creatinine and liver enzyme levels, it eliminated the effect of cisplatin on leucocyte counts. Cisplatin increased significantly urinary creatinine level and creatinine clearance. Cisplatin caused some histological changes in kidney and liver.

Effects of ciprofloxacin on chromosomes, and hepatic and renal functions in rats.

In the present study, ciprofloxacin, a fluoroquinolone antibiotic, was evaluated in vivo. The toxic effects of ciprofloxacin (20 and 200 mg/kg/day) on kidney and liver functions, and on chromosomes were investigated in rats. 20 mg/kg/day ciprofloxacin did not significantly affect urine composition, serum enzyme levels and the haematocrit, affecting only creatinine clearance. The 200 mg/kg/day dose increased urine volume, serum creatinine and and creatinine clearance. Ciprofloxacin at both doses had no significant numerical effect on chromosomes, but there was a statistically significant dose-dependent increase in chromatid-type breakage. Significant histological changes in the liver and kidney were not observed.