Investigation of the relationship between nitric oxide metabolites' levels and adenosine deaminase activity in 7,12-dimethylbenz[a]anthracene induced mouse liver

7,12-Dimethylbenz[a]anthracene (7,12-DMBA) is a member of the polycyclic aromatic hydrocarbons with a severe carcinogenic effect. In this study, nitrate levels and ADA (Adenosine deaminase) activity in the liver homogenates of mice were measured and the effect of free radicals induced by 7,12-DMBA on inducible nitric oxide synthase (iNOS) and ADA activity were investigated. Antioxidant effects of melatonin were also compared. Three groups of mice were included in the study. The first served as control, the second was treated only with 7,12-DMBA and the third was treated with 7,12-DMBA + melatonin. Spectrophotometric methods were used at all measurements. Data were analyzed using Kruskal-Wallis Variance Analysis Test and Mann-Whitney U Test that were applied to the groups. The nitrate concentrations of mouse liver were as follows: 4.98 +/- 0.63 micro mol/L in the control group (n = 10), 8.23 +/- 1.58 micro mol/L (higher than control group, p < 0.05) in the 7,12-DMBA-treated group (n = 10), and 6.43 +/- 0.57 micro mol/L (lower than 7,12-DMBA-treated group, p < 0.05) in the 7,12-DMBA + melatonin-treated group (n = 10). Liver ADA activities were measured to be 4.14 +/- 0.674 U/L in the control group, 6.25 +/- 1.261 U/L (higher than control group, p < 0.05) in the 7,12-DMBA-treated group, and 4.93 +/- 0.916 U/L (lower than 7,12-DMBA-treated group, p < 0.05) in the 7,12-DMBA+melatonin-treated group. Differences between free nitrite levels were no significantly. Results demonstrated that nitrate levels and ADA activities were increased by means of free radicals induced by 7,12-DMBA. Melatonin inhibited the 7,12-DMBA induced increase that was observed in the activities of ADA enzyme and nitrate levels. It is concluded that determination of ADA activity and nitrate levels can be useful in the assessment of liver damage caused by toxic chemicals.     

Depurination of ribosomal RNA and inhibition of viral RNA translation by an antiviral protein of Celosia cristata

An antiviral protein (25 kD) isolated from leaves of Celosia cristata (CCP 25) was tested for depurination study on ribosomal RNA from yeast. Ribosomal RNA yielded 360 nucleotide base fragment after treatment with CCP 25 indicating that CCP 25 was a ribosome inactivating protein. CCP 25 also inhibited translation of brome mosaic virus (BMV) and pokeweed mosaic virus (PMV) RNAs in rabbit reticulocyte translation system. The radioactive assay showed that incorporation of [35S]-methionine was less in translation proteins of BMV nucleic acid when CCP 25 was added to translation system. This indicated that antiviral protein from Celosia cristata not only depurinated ribosomal RNA but also inhibited translation of viral RNA in vitro.     

A non-toxic pokeweed antiviral protein mutant inhibits pathogen infection via a novel salicylic acid-independent pathway

Pokeweed antiviral protein (PAP), a ribosome-inactivating protein isolated from Phytolacca americana, is characterized by its ability to depurinate the sarcin/ricin (S/R) loop of the large rRNA of prokaryotic and eukaryotic ribosomes. In this study, we present evidence that PAP is associated with ribosomes and depurinates tobacco ribosomes in vivo by removing more than one adenine and a guanine. A mutant of pokeweed antiviral protein, PAPn, which has a single amino acid substitution (G75D), did not bind ribosomes efficiently, indicating that Gly-75 in the N-terminal domain is critical for the binding of PAP to ribosomes. PAPn did not depurinate ribosomes and was non-toxic when expressed in transgenic tobacco plants. Unlike wild-type PAP and a C-terminal deletion mutant, transgenic plants expressing PAPn did not have elevated levels of acidic pathogenesis-related (PR) proteins. PAPn, like other forms of PAP, did not trigger production of salicylic acid (SA) in transgenic plants. Expression of the basic PR proteins, the wound-inducible protein kinase and protease inhibitor II, was induced in PAPn-expressing transgenic plants and these plants were resistant to viral and fungal infection. These results demonstrate that PAPn activates a particular SA-independent, stress-associated signal transduction pathway and confers pathogen resistance in the absence of ribosome binding, rRNA depurination and acidic PR protein production.     

The effects of chronic smoking on lung tissue and the role of alpha lipoic acid

Abstract

We investigated the effects of alpha lipoic acid (ALA) on blood and lung tissue exposed chronically to cigarette smoke (CS). Female Sprague-Dawley rats were divided into three groups. Group 1 was the control group (CON): fresh air was supplied twice daily and 0.1 ml physiological saline was given orally for 8 weeks. Group 2 was exposed to CS: 12 cigarettes were smoked daily at two sessions for 1 h and 0.1 ml saline was given orally for 8 weeks. Group 3 (CS + ALA) was exposed to 12 cigarettes daily in two sessions for 1 h and 100 mg/kg/day ALA was given orally for 8 weeks. DNA damage was assessed using comet analysis; oxidative damage was assessed using ischemia-modified albumin (IMA) from blood; and total oxidant status (TOS), total antioxidant status (TAS) and oxidative stress index (OSI) were measured in blood and lung tissue. Histopathological and immunohistochemical evaluation of hypoxia-inducible factor (HIF)-1α, and ?2α, caspase-3, vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF₂) were conducted using lung tissue. The oxidative markers, TOS, OSI and IMA, and the comet analysis score were increased and the TAS level was decreased in the blood of the CS group compared to the CON group. IMA levels in blood, and TOS and OSI levels in the lung were decreased significantly in the CS + ALA group compared to the CS group. We observed increased septal wall thickness, marked and diffuse inflammatory reaction, emphysema, and necrotic cell debris in bronchial and bronchiolar lumens in the CS group. HIF-1α, HIF-2α, caspase-3 and FGF₂ expressions were increased, while VEGF expression decreased in the lung tissues of the CS group compared to the CON group. ALA slightly ameliorated the damage caused by chronic exposure to CS in the lungs, but further investigation is needed to determine its possible protective effects at different dosages.     

The effect of temperature on photosynthetic induction under fluctuating light in Chrysanthemum morifolium

The photosynthetic response was investigated on Chrysanthemum morifolium under dynamic light conditions in the 20–35 °C temperature range to evaluate the effect of climatic variables on photosynthetic induction. The plant material was grown under uniform, controlled conditions and its gas exchange was analyzed. The gas exchange measurements were used to investigate the rate of induction, momentary induction state, and the opening of stomata. At the varying temperature ranges and under dynamic light conditions, C. morifolium reached a quasi-steady-state induction equilibrium (IS_eq(PAR,T)) within 14–45 min. For the same level of photosynthetically active radiation (PAR), the equilibrated level of steady-state induction increased as the temperature increased. It was highest approximately at 30 °C. The induction state was equilibrated at a lower level as the temperature increased to 35 °C. The interaction effect of PAR and temperature on induction state was not significant. The rate of photosynthetic induction and the time required at which the induction reached its 90 % value (t ₉₀) was influenced by PAR significantly. The light history of a leaf had a significant effect on t ₉₀, indicating that the time to reach a steady-state induction is different depending on the light environment and the period at which the leaf was exposed to light. The velocity of the photosynthetic induction was not affected by the temperature. It was associated with stomatal conductance of the leaf prior to the onset of light (g _Sini).     

HER2 Ile655Val and PTEN IVS4 polymorphisms in patients with breast cancer

Although HER2/PTEN pathway is commonly disrupted in cancer, association of HER2 and PTEN polymorphisms with breast cancer (BC) remains controversial. We investigated the HER2 Ile655 Val and PTEN IVS4 polymorphisms in patients with BC in Turkish population. HER2 Ile655Val (rs 1136201) and PTEN IVS4 (rs 3830675) polymorphisms were determined using polymerase chain reaction-based restriction fragment length polymorphism (PCR–RFLP) in blood samples of 118 BC patients and 118 age-matched healthy controls. We found that the frequency of the Ile/Val genotype of HER2 Ile655Val gene was significantly higher in BC patients (p < 0.009; OR: 1,983 95 % CI: 1.181—3.328). The presence of ATCTT insertion (+/+) genotype at downstream of exon 4 in intron 4 of PTEN IVS4 gene was also associated with 1.83 fold decreased risk of BC development (p < 0.033; OR: 1.83, 95 % CI: 1.11—3.03). Analysis on clinico-pathological parameters showed neither HER2 Ile655Val nor PTEN IVS4 genotypes were not associated with any of the variables (p > 0.05).In conclusion, our findings suggest that the Ile/Val genotype of HER2 and ATCTT insertion (+/+) genotype of PTEN IVS4 gene may play an important role as genetic markers for breast cancer risk, but both genes genotypes may not be useful for predicting tumor prognosis in Turkish population.     

The effects of age and gender on the relationship between HMGCR promoter-911 SNP (rs33761740) and serum lipids in patients with coronary heart disease

Background

Hydroxymethylglutaryl-Coenzyme A Reductase (HMGCR) catalyzes the rate-limiting step of cholesterol biosynthesis. This enzyme is the target of the widely available cholesterol lowering statins. In this population-based case–control study, the frequencies of -911 C>A polymorphism (rs3761740) of the HMGCR gene in patients with coronary heart disease (CHD) and healthy subjects were investigated and the correlations between the different genotypes and hypercholesterolemia with cardiovascular risk factors were analyzed.

Methods

The HMGCR genotypes were determined in 365 patients with CHD and 365 controls by PCR–RFLP assay. Anthropometric measurements were measured in all participants.

Results

There was no significant difference in the genotype frequencies of the HMGCR polymorphism between the male subjects of both patient and control groups, however, the HMGCR-CC genotype was found to be more frequent in female patients with CHD than female controls (p = 0.002). The HMGCR-CC genotype showed higher total-cholesterol (TC) and LDL-cholesterol (LDL-C) levels than the CA + AA genotypes in male CHD patients (p = 0.018). Due to this significant sex interaction, a multivariate analysis was conducted on the patient group. In the multivariate logistic regression analysis, the HMGCR-CC genotype was significantly associated with age < 55 (OR = 2.837, p = 0.001) and TC ≥ 5.18 mmol/L (OR = 1.970, p = 0.027) in male subjects. However, this association was not observed in female patients (p > 0.05). This analysis confirmed that the HMGCR-CC genotype was associated with elevated TC levels in male CHD patients with age < 55 years.

Conclusion

These results suggest that age and sex modify the contribution of the HMGCR-911 polymorphism to fasting serum TC, LDL-C levels and risk of CHD.