The transcription factor Forkhead Box P3 gene variants affect idiopathic recurrent pregnancy loss.

INTRODUCTION: Forkhead Box P3 (FOXP3) is an essential transcription factor for the induction and development of Tregs. It plays an important role in regulation and suppression of immune responses. We tested whether FOXP3 gene variants are associated with idiopathic recurrent miscarriages (IRM). METHODS: We included 200 women with at least three unexplained spontaneous abortions before twentieth week of gestation and 300 healthy parous women. The detection of genetic variants of rs2232365, and rs5902434 SNPs were carried-out by using polymerase chain reaction (PCR) with sequence-specific primers, while rs3761548 and rs2294021 SNPs were genotyped by PCR followed by RFLP analysis. The logistic odds ratios (ORs) of idiopathic RM risk were estimated with a 95% confidence interval (CI) after maternal age adjustment. Multifactor dimension reduction (MDR) analysis was used to evaluate the potential SNP ∼ SNP interactions. RESULTS: Single marker analysis revealed an increased risk ranged from almost 3-fold-2-fold for rs2232365, rs3761548, rs5902434 and rs2294021 SNPs in IRM cases. The mutant haplotype carriers of rs2232365, rs3761548, rs5902434 and rs2294021 SNPs showed an increased risk of 2.5-fold for IRM cases. Linkage disequilibrium analysis revealed moderate LD between rs2232365, rs3761548, rs5902434 and rs2294021 SNPs. The MDR analysis revealed 6-fold increased risk for IRM cases in four factor models of rs2232365, rs3761548, rs5902434 and rs2294021 SNPs. The maximum testing accuracy, highest cross validation consistency and greater significance was observed in four SNP model. DISCUSSION: These results suggest that variants of FOXP3 SNPs namely; rs2232365, rs3761548, rs5902434 and rs2294021 may be associated with idiopathic RM.

Association of HLA-G promoter and 14-bp insertion-deletion variants with acute allograft rejection and end-stage renal disease.

The aim of this study was to investigate the HLA-G 14-bp insertion/deletion (I/D) polymorphism among end-stage renal disease (ESRD) patients. Cytomegalovirus (CMV) infection, acute allograft rejection (AR) and overall survival after renal transplantation was investigated in 300 ESRD patients and 302 age, sex and ethnicity-matched controls. Sequencing was performed to evaluate the impact of HLA-G promoter region single-nucleotide polymorphisms (SNPs) whereas semi-quantitative PCR method was used to determine the probable HLA-G expression pattern among ESRD and AR cases. Further, soluble human leukocyte antigen (HLA)-G (sHLA-G) expression levels were compared in AR vs non-AR cases in the light of HLA-G 14-bp I/D polymorphism. Increased risk was found for 14-bp D/D (deletion-DD) genotype and 14-bp D allele [DD: odds ratio (OR) = 1.46, 95% confidence interval (CI) = 1.03-2.06, P value = 0.0358; D: OR = 1.29, 95% CI = 1.03-1.62, P value = 0.0277], respectively for ESRD and CMV infection (DD: OR = 2.70, 95% CI = 1.45-5.05, P value = 0.0021; D: OR = 1.94, 95% CI = 1.22-3.08, P value = 0.0052). Nearly fourfold (OR = 3.62, 95%CI = 1.61-8.14, p = 0.0039) risk was observed for 14-bp I/I (insertion-II) genotype for AR. Survival analysis showed increased overall survival (OS) (AR or death) for 14-bp D/D genotype. HLA-G promoter region sequencing was carried out among 60 ESRD patients and 100 normal controls which showed increased risk for -964 G>A, -725 C>G/T and -486 A>C SNPs. -964 G>A and -725 C>G/T SNPs showed risk association for AR patients. High level of HLA-G transcripts was observed among non-AR patients. Further soluble HLA-G (sHLA-G) showed increased levels in ESRD patients (mean ± SEM; 62.16 ± 2.43 U/ml) as compared to controls (mean ± SEM; 21.06 ± 3.89 U/ml) (P = <0.0001). The 14-bp I/I, 14-bp I/D and 14-bp D/D genotypes showed significantly higher levels of sHLA-G among non-AR as compared to AR patients.

Role of l-Arginine on Dyslipidemic Conditions of Acute Myocardial Infarction Patients.

Oxidative stress conditions associated with atherosclerosis leads to oxidative modification of low-density lipoprotein (LDL). The body's capabilities to inhibit LDL oxidation and to remove or neutralize the atherogenic oxidized LDL (ox-LDL) are limited. When the LDL cholesterol level increases in the blood, it leads to dangerous consequences like atherosclerosis, leading to myocardial infarction. The major effect of an antioxidant in the LDL environment is to prevent the formation of ox-LDL (during atherogenesis. Strategies to reduce LDL oxidation and prevent atherogenesis can involve the enrichment of arterial cells with potent antioxidants that can prevent oxidative damage to the arterial wall. The objective of this study is to evaluate the effect of l-arginine on serum lipid and cholesterol levels in the patients of acute myocardial infarction (AMI). The study consisted of 70 AMI patients and 60 healthy individuals (serving as control) age 55-65 years. Serum levels of total cholesterol, high density lipoprotein (HDL), LDL and Triglycerides were determined on day 1 and day 15 of l-arginine administration (oral dose 3 g/day). The total cholesterol/HDL and the LDL/HDL ratio were calculated and compared. As per the observations, l-arginine administration was found to improve the lipid profile of the subjects. Hence it could be used as an adjuvant therapy for AMI and as a preventive measure for the onset of the disease in the healthy elderly also.

Assessment of free radical-mediated damage in head and neck squamous cell carcinoma patients and after treatment with radiotherapy.

Reactive oxygen species (ROS) produced as a part of cellular metabolism can interact with biological macromolecules such as DNA, proteins and lipids and interfere with their normal functions, leading to the loss of cellular viability. ROS have been implicated in many pathophysiological conditions including cancer. In the present study, the damage caused by ROS and the effect of radiation in head and neck squamous cell carcinoma (HNSCC) patients were assessed in the erythrocytes by analyzing the superoxide dismutase (SOD) and catalase (CAT) activities, and levels of total thiols (T-SH) and malondialdehyde (MDA, a marker for lipid peroxidation). Blood samples were collected before the start of treatment and after the completion of radiotherapy. Both SOD and CAT activities were decreased in untreated patients, but elevated in patients after treatment. The T-SH levels were also depleted in untreated HNSCC patients, but elevated non-significantly after radiation therapy (p>0.05). The levels of MDA showed a significant increase in both untreated patients and after radiation therapy when compared with normal subjects (p<0.05). Thus, the present study indicated that the free radical-mediated damage was aggravated in untreated HNSCC patients, but the levels of antioxidants returned to baseline or nearly so after the treatment with radiation therapy.

Protective Role of l-Arginine Against Free-Radical Mediated Oxidative Damage in Patients with Unstable Angina.

Unstable angina is a critical condition of heart resulting from narrowing of vessels supplying blood to heart. Ischemia of the myocardium leads to oxidative stress and severe tissue damage. The objective of the present study was to determine the effect of l-arginine administration on the oxidant-antioxidant homeostasis which otherwise gets imbalanced in patients with cardiovascular diseases. The results obtained, show improvement in the oxidant-antioxidant levels of the subjects upon incorporation of l-arginine. Our findings suggest that supplementation of l-arginine along with regular anti-anginal therapy may be beneficial to the patients of unstable angina.

Therapeutic role of L-arginine on free radical scavenging system in ischemic heart diseases.

Increased production of free radicals under oxidative stress conditions plays a vital role in the impairment of endothelial function and also in the pathogenesis of ischemic heart diseases. Ischemia, followed by reperfusion, leads to the exacerbated formation of oxy- free radicals. These reactive oxygen species through a chain of reactions damage the cardiomyocytes and cause more injury to the myocardium. L-Arginine is reported to act as free radical scavenger, inhibits the activity of pro-oxidant enzymes and thus acts as an antioxidant and these roles of L-arginine are mediated by nitric oxide (NO). In the present study, the effect of oral administration of L-arginine (3 g/day for 7 days) on some antioxidant enzymes, total thiols, lipid peroxidation measured as malondialdehyde (MDA), and plasma ascorbate levels in myocardial ischemic patients was investigated. We observed an increase in the activity of superoxide dismutase (SOD), total thiols (T-SH) and plasma ascorbate levels and a decrease in the activity of xanthine oxidase (XO), MDA levels, carbonyl content and serum cholesterol in the patients on oral administration of L-arginine. The present study demonstrates that L-arginine administration may be beneficial to patients with myocardial ischemic disorders, such as acute myocardial infarction and acute angina.

Xanthine oxidase as a marker of myocardial infarction.

In the present communication, we report remarkably elevated levels of xanthine oxidase activity in the blood of the patients with myocardial infarction when compared to age and sex matched healthy persons. Highly significant increase of malondialdehyde, serving as an index of lipid peroxidation and thus free radical mediated damage, has also been found in the patients. We propose the measurement of the blood levels of xanthine oxidase, a very simple, reliable and less time consuming method as an indicator of myocardial infarction.

Adenosine deaminase in ischemia reperfusion injury in patients with myocardial infarction.

A comparative study on the levels of erythrocyte adenosine deaminase and lipid peroxidation has been undertaken in patients with myocardial infarction before and after thrombolysis along with matched healthy individuals. Our findings show that adenosine deaminase activity is highly elevated in post-reperfused patients when compared to pre- thrombolysed and healthy persons. Malondialdehyde(MDA) levels are also significantly increased in post-thrombolysed patients. The study reveals an important role of adenosine deaminase in reperfusion injury in patients with myocardial infarction.

Effect of ascorbic acid supplementation on certain oxidative stress parameters in the post reperfusion patients of myocardial infarction.

Reperfusion injury causes oxidative stress thereby resulting in an imbalance between oxidant-antioxidant systems. In the present communication, the effect of ascorbic acid supplementation has been studied on certain oxidant and antioxidant parameters in the blood of the patients with myocardial infarction before and after thrombolysis. In patients after thrombolysis, the activity of antioxidant enzyme, superoxide dismutase, in the blood was found to be significantly reduced where as the activity of the oxidant enzyme, xanthine oxidase, was found to be significantly increased. Malondialdehyde levels, the index of free radical mediated damage, was also found to be significantly elevated in thrombolysed patients compared to the patients before thrombolysis. Supplementation of vitamin C to the post reperfusion patients restored these parameters back to normal or near normal levels.

Status of some free radical scavenging enzymes in the blood of myocardial infarction patients.

Pro-oxidant and anti-oxidant systems and their levels have significant roles in occlusive vascular diseases. In the present communication, we have measured the levels of some representative anti-oxidant enzymes in the blood of the patients of myocardial infarction after reperfusion and compared them to age and sex matched healthy persons. Our findings show that the activities of anti-oxidant enzymes (viz. SOD, catalase and glutathione reductase) are significantly decreased whereas there is significant increase in the levels of malonaldialdehyde (a marker of free radical-mediated damage) in the patients. The findings point out that ischemic myocardial disorders are associated with excessive free radical generation and free radical-mediated damage of lipids.

Oxidative stress parameters in erythrocytes of post-reperfused patients with myocardial infarction.

The effect of reperfusion of patients with myocardial infarction on the levels of some anti-oxidant enzymes, total thiols, malondialdehyde formation in erythrocytes and plasma ascorbate levels have been investigated. Significantly decreased activities of catalase and superoxide dismutase and decreased levels of total thiols in RBC's and ascorbic acid in plasma suggest that reperfusion of the infarcted myocardium leads to oxidative stress conditions wherein anti-oxidant mechanisms become less effective in coping with the oxidative insult. This view is further supported by the observation that in the post reperfused patients there is a highly significant enhancement in the levels of malondialdehyde.

Effect of vitamin E on platelet enzymatic anti-oxidants in the patients of myocardial infarction.

Effect of administration of 600 mg. vitamin E each day, for six days, was observed on activity of some of the anti-oxidant enzymes and levels of malondialdehyde (as an index of free radical mediated damage) in the platelets of patients reperfused after myocardial infarction. It has been found that vitamin E administration significantly lowers the level of malondialdehyde in the patients. Vitamin E administration increases the activities of anti oxidant enzymes (viz. superoxide dismutase, glutathione reductase and catalase) tested both in the patients and healthy controls. Vitamin E administration causes general stimulation of anti-oxidant enzyme activities both in healthy persons and the patients, however, lowering of lipid per-oxidation upon administration of vitamin E is specific for patients. These findings exhibit beneficial role of vitamin E administration in the management of the patients reperfused after myocardial infarction.

Effect of vitamin E on the platelet xanthine oxidase and lipid peroxidation in the patients of myocardial infarction.

Platelets play important role in precipitating ischaemic myocardial syndromes in many ways. One of the consequences of ischaemic diseases is excessive generation of oxygen derived free radicals that have numerous pathophysiological consequences. Platelet pro-oxidant enzyme, xanthine oxidase is one of the sources of generation of free radicals. In the present paper, we report the effect of administration of vitamin E along with aspirin on the levels of platelet xanthine oxidase and extent of free radical mediated damage in the patients reperfused after myocardial infarction.Our findings show that administration of 400 mg. vitamin E for six days along with 80 mg. aspirin has an excellent anti-oxidant effect as evidenced by reduced platelet xanthine oxidase activity and lowering of malondialdehdye (MDA) levels which is an index of the extent of free radical mediated damage.

Broad identification of bacterial type from pus by 1H MR spectroscopy.

BACKGROUND: Abscess formation is a common bacterial infection and requires an immediate antimicrobial approach for apposite treatment. Delay in patient treatment is usually a common feature, as the bacterial identification of clinical samples is based on the culture, which is a time-consuming exercise. The current study was aimed at developing an alternative technique with the potential for rapid bacterial group identification. MATERIALS AND METHODS: In the present study we performed ex vivo proton magnetic resonance spectroscopy of 40 pus samples collected from abscesses in different locations and the results have been retrospectively compared with the microorganism identified in the pus culture. In addition, the microbes obtained from the culture have been further subcultured and studied with magnetic resonance spectroscopy to identify the bacterial fingerprint in the pus sample seen on spectroscopy. RESULTS: On reviewing the spectra obtained from the various abscesses, they were found to be qualitatively similar for a particular bacterium. The similar spectral pattern of the pus with obligate aerobes/anaerobes and pure cultures of the same bacteria suggests its strict metabolism under in vivo and in vitro conditions, respectively. CONCLUSIONS: The characteristic metabolite pattern of obligate anaerobes may be used as a prototype for its rapid identification. This information may be of value for more appropriate clinical management of such cases.

Reversible suppression of nitric oxide system in essential hypertension.

Despite enormous research in the field of hypertension, its pathophysiology still remains largely unresolved and appears to be multifactorial. In the present communication, we have analyzed the status of nitric oxide (NO) in the patients with essential hypertension and age matched controls. We have found that the levels of NO are lowered in essential hypertension. The normalization of blood pressure by administration of antihypertensive therapy causes rise in the NO level indicating that perturbed NO status in essential hypertension is reversible. Addition of antioxidant to the antihypertensive drugs causes a further, though non significant, rise in the levels of NO, suggesting that antioxidants may be combined with antihypertensive drugs as adjunct in the management of essential hypertension.

Protective effect of green tea extract against the erythrocytic oxidative stress injury during mycobacterium tuberculosis infection in mice.

The present study has been undertaken to monitor the extent of oxidative stress in mice infected with M tuberculosis and the role of crude green tea extract in repairing the oxidative damage. The mice were divided into three groups of 9 each; normal, infected-untreated and infected-treated. The infected group of animals exhibited significant enhancement of erythrocytic catalase and glutathione peroxidase activities along with elevated levels of erythrocytic total thiols and plasma lipid peroxidation as compared to normal animals. The infected group also exhibited significantly decreased activity of superoxide dismutase and levels of glutathione in erythrocytes. Upon oral administration of green tea extract for seven days the oxidative stress parameters were reverted back to near normal levels as evidenced by a fall in catalase, glutathione peroxidase, total thiol and extent of lipid peroxidation with concomitant increase in the levels of SOD and reduced glutathione in infected animals. The findings thus, portray that there is a high oxidative stress during early stages of tuberculosis and antioxidants such as green tea extract, can play a vital role by reducing stress through adjuvant therapy.

Molecular mechanisms of DNA damage and repair: progress in plants.

Despite stable genomes of all living organisms, they are subject to damage by chemical and physical agents in the environment (e.g., UV and ionizing. radiations, chemical mutagens, fungal and bacterial toxins, etc.) and by free radicals or alkylating agents endogenously generated in metabolism. DNA is also damaged because of errors during its replication. The DNA lesions produced by these damaging agents could be altered base, missing base, mismatch base, deletion or insertion, linked pyrimidines, strand breaks, intra- and inter-strand cross-links. These DNA lesions could be genotoxic or cytotoxic to the cell. Plants are most affected by the UV-B radiation of sunlight, which penetrates and damages their genome by inducing oxidative damage (pyrimidine hydrates) and cross-links (both DNA protein and DNA-DNA) that are responsible for retarding the growth and development. The DNA lesions can be removed by repair, replaced by recombination, or retained, leading to genome instability or mutations or carcinogenesis or cell death. Mostly organisms respond to genome damage by activating a DNA damage response pathway that regulates cell-cycle arrest, apoptosis, and DNA repair pathways. To prevent the harmful effect of DNA damage and maintain the genome integrity, all organisms have developed various strategies to either reverse, excise, or tolerate the persistence of DNA damage products by generating a network of DNA repair mechanisms. A variety of different DNA repair pathways have been reported that include direct reversal, base excision repair, nucleotide excision repair, photoreactivation, bypass, double-strand break repair pathway, and mismatch repair pathway. The direct reversal and photoreactivation require single protein, all the rest of the repair mechanisms utilize multiple proteins to remove or repair the lesions. The base excision repair pathway eliminates single damaged base, while nucleotide excision repair excises a patch of 25- to 32-nucleotide-long oligomer, including the damage. The double-strand break repair utilizes either homologous recombination or nonhomologous endjoining. In plant the latter pathway is more error prone than in other eukaryotes, which could be an important driving force in plant genome evolution. The Arabidopsis genome data indicated that the DNA repair is highly conserved between plants and mammals than within the animal kingdom, perhaps reflecting common factors such as DNA methylation. This review describes all the possible mechanisms of DNA damage and repair in general and an up to date progress in plants. In addition, various types of DNA damage products, free radical production, lipid peroxidation, role of ozone, dessication damage of plant seed, DNA integrity in pollen, and the role of DNA helicases in damage and repair and the repair genes in Arabidopsis genome are also covered in this review.

Enzymatic oxidant and antioxidants of human blood platelets in unstable angina and myocardial infarction.

In ischaemic heart conditions we report a remarkable increase in platelet xanthine oxidase activity and rise in the levels of malondialdehyde (MDA) with concomitant decrease in the activities of free radical scavenging enzymes - superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The increased levels of free radical generating system and MDA and lowered levels of free radical scavenging systems seem to have critical role in ischaemic heart conditions.