Evaluating the effect of a mixture of two main conjugated linoleic acid isomers on hepatic steatosis in HepG2 cellular model.

Hepatic steatosis is an early form of non-alcoholic fatty liver disease (NAFLD), caused by abnormal fat deposition in the hepatocytes. Conjugated linoleic acid (CLA) is a group of positional and geometric dienoic isomers of linoleic acid that attract significant attention because of its beneficial effects on chronic diseases such as cancer, obesity, and metabolic syndrome. This study examined the influence of a mixture of two main CLA isomers (CLA-mix) on lipid accumulation and lipid metabolism-related genes using HepG2 cells treated with palmitic acid (PA) as an in vitro model for hepatic steatosis. Methods and Results: HepG2 cells were treated for 24 h: control (BSA), model (BSA + PA), and treated groups (BSA-PA + non-toxic concentrations of CLA-mix). Intracellular lipid deposition, triglyceride (TG), total cholesterol (TC) and gene expression were measured by Oil-Red O staining, colorimetric assay kits and real-time PCR, respectively. CLA-mix at high concentrations had significantly decreased intracellular total lipid and TG deposition compared to the model group. However, none of the CLA-mix concentrations had a significant effect on the intracellular TC level. CLA-mix significantly increased the expression of some genes mainly regulated by PPARα but did not alter the expression of lipogenesis-related genes. Conclusions: These results demonstrate that high concentrations of CLA-mix protect against hepatic steatosis and play a role in regulating fatty acid oxidation and bile excretion through the PPARα pathway. It is suggested that the effect of different ratios of two main CLA isomers on the amount and ratio of bile compounds be investigated in future studies.

Generation of a Transgenic Zebrafish Model for Pancreatic Beta Cell Regeneration.

BACKGROUND: Diabetes is a major worldwide health problem. It is widely accepted that the beta cell mass decreases in type I diabetes (T1D). Accordingly, beta cell regeneration is a promising approach to increase the beta cell mass in T1D patients. However, the underlying mechanisms of beta cell regeneration have yet to be elucidated. One promising avenue is to create a relevant animal model to explore the underlying molecular and cellular mechanisms of beta cell regeneration. The zebrafish can be considered a model in beta cell regeneration studies because the pancreas structure and gene expression pattern are highly conserved between human and zebrafish. MATERIALS AND METHODS: In this study, the Tol2 transposase was exploited to generate a Tg(Ins:egfp-nfsB) zebrafish model that expressed a fusion protein composed of enhanced green fluorescent protein (EGFP) and nitroreductase (NTR) under control of the Ins promoter. RESULTS: Metronidazole (MTZ) treatment of Tg(ins:egfp-nfsB) zebrafish larvae led to selective ablation of beta cells. Proof-of-concept evidence for beta cell regeneration in the transgenic larvae was observed two days after withdrawal of MTZ. CONCLUSION: This study suggests that the Tg(ins:egfp-nfsB) zebrafish can be used as a disease model to study beta cell regeneration and elucidate underlying mechanisms during the regeneration process.

Association of carotid intima media thickness with atherogenic index of plasma, apo B/apo A-I ratio and paraoxonase activity in patients with non-alcoholic fatty liver disease.

BACKGROUND: NAFLD patients have higher risk of atherosclerosis cardiovascular disease (ASCVD). apo B/apoA-I ratio and atherogenic index of plasma (AIP) have been suggested as biomarker for ASCVD. METHODS: apo A-I, apoB, aryl esterase (ARE) and paraoxonase (PON) activities and carotid intima media thickness (cIMT) were determined in 49 NAFLD patients and 33 controls. RESULTS: Plasma levels of apo A-I, adiponectin, ARE and PON activities decreased in NAFLD patients, while apo B, AIP and apoB/apo A-I ratio level were higher in NAFLD patients compared to controls. Furthermore, cIMT showed a positive association with AIP, apo B/apo A-I ratio and AIP + (apo B/apo A-I) in NAFLD patients. Strikingly, AIP + (apo B/apo A-I) showed a good ability to discriminating increased cIMT in NAFLD patients. CONCLUSIONS: The result showed that AIP and apo B/apo A-I associated with cIMT in NAFLD patients; however, more study are needed to prove this concept.

Increased inflammatory responsiveness of peripheral blood mononuclear cells (PBMCs) to in vitro NOD2 ligand stimulation in patients with ankylosing spondylitis.

Background: Ankylosing spondylitis (AS) is a common debilitating rheumatic disease in which the innate immune components especially the Interleukin (IL)-23/IL-17 axis related genes play important role in its pathogenesis. Nucleotide binding oligomerization domain-containing protein (NOD)2, as an innate receptor, is critical for IL-23 production in cells. Therefore, we aimed to stimulate NOD2 signaling and study its effects on cytokine production in peripheral blood mononuclear cells (PBMC) of these patients. Methods: PBMCs from 18 patients with active AS and 18 healthy individuals were separated by Ficoll-Hypaque density gradient centrifugation and cultured in the presence of muramyl dipeptide (MDP), as NOD2 ligand. Quantitative expression analysis of NOD1, NOD2, RIPK2, SLC15A4, NLRP1, NLRP3, IL23A, IL17A, IL1B, and TNFA genes was performed using Real-time polymerase chain reaction (PCR). Finally, protein changes of IL23A and IL17A expression were validated using enzyme linked immunosorbent assay (ELISA). Results: Apart from NOD1 that tend to be downregulated in the controls, all the selected genes showed overexpression in response to MDP in cells from the studied groups. Except RIPK2, all the genes had higher expression changes upon MDP stimulation in the AS population. Overexpression of IL23A and IL17A were confirmed at protein levels using ELISA. The strong positive correlation between NLRP3 and NOD2 was decreased after stimulation but new correlations between NLRP3 and IL1B, RIPK2 and SLC15A4 were observed after treatment. Conclusions: This study indicated that AS PBMCs were hyper-responsive to MDP stimulation. This observation implies an important role of NOD2 in the pathogenesis of inflammatory diseases including AS.

Impaired HDL cholesterol efflux capacity in patients with non-alcoholic fatty liver disease is associated with subclinical atherosclerosis.

Non-alcoholic fatty liver disease (NAFLD) is associated with a substantial increased risk of atherosclerotic cardiovascular disease (ASCVD), which is partly related to dyslipidemia and low HDL-C level. The cardioprotective activity of HDL in the body is closely connected to its role in promoting cholesterol efflux, which is determined by cholesterol efflux capacity (CEC). Hitherto, the role of HDL, as defined by CEC has not been assessed in NAFLD patients. In this research study, we present the results of a study of cAMP-treated J774 CEC and THP-1 macrophage CEC in ApoB-depleted plasma of 55 newly diagnosed NAFLD patients and 30 controls. Circulating levels of ApoA-I, ApoB, preß-HDL, plasma activity of CETP, PLTP, LCAT and carotid intima-media thickness (cIMT) were estimated. cAMP-treated J774 and THP-1 macrophage CEC were found to be significantly lower in NAFLD patients compared to controls (P < 0.001 and P = 0.003, respectively). In addition, it was discovered that both ApoA-I and preß1-HDL were significantly lower in NAFLD patients (P < 0.001). Furthermore, cAMP-treated J774 CEC showed independent negative correlation with cIMT, as well as the presence of atherosclerotic plaque in NAFLD patients. In conclusion, our findings showed that HDL CEC was suppressed in NAFLD patients, and impaired cAMP-treated J774 CEC was an independent risk factor for subclinical atherosclerosis in NAFLD patients, suggesting that impaired HDL functions as an independent risk factor for atherosclerosis in NAFLD.

Evaluation of the effects of various sound pressure levels on the level of serum aldosterone concentration in rats.

INTRODUCTION: Noise exposure may have anatomical, nonauditory, and auditory influences. Considering nonauditory impacts, noise exposure can cause alterations in the automatic nervous system, including increased pulse rates, heightened blood pressure, and abnormal secretion of hormones. The present study aimed at examining the effect of various sound pressure levels (SPLs) on the serum aldosterone level among rats. MATERIALS AND METHODS: A total of 45 adult male rats with an age range of 3 to 4 months and a weight of 200 ± 50 g were randomly divided into 15 groups of three. Three groups were considered as the control groups and the rest (i.e., 12 groups) as the case groups. Rats of the case groups were exposed to SPLs of 85, 95, and 105 dBA. White noise was used as the noise to which the rats were exposed. To measure the level of rats' serum aldosterone, 3 mL of each rat's sample blood was directly taken from the heart of anesthetized animals by using syringes. The taken blood samples were put in labeled test tubes that contained anticoagulant Ethylenediaminetetraacetic acid. In the laboratory, the level of aldosterone was assessed through Enzyme-linked immunosorbent assay protocol. The collected data were analyzed by the use of Statistical Package for Social Sciences (SPSS) version 18. RESULTS: The results revealed that there was no significant change in the level of rats' serum aldosterone as a result of exposure to SPLs of 65, 85, and 95 dBA. However, the level of serum aldosterone experienced a remarkable increase after exposure to the SPL of 105 dBA (P < 0.001). Thus, the SPL had a significant impact on the serum aldosterone level (P < 0.001). In contrast, the exposure time and the level of potassium in the used water did not have any measurable influence on the level of serum aldosterone (P = 0.25 and 0.39). CONCLUSION: The findings of this study demonstrated that serum aldosterone can be used as a biomarker in the face of sound exposure.

Modeling signal-to-noise ratio of otoacoustic emissions in workers exposed to different industrial noise levels.

INTRODUCTION: Noise is considered as the most common cause of harmful physical effects in the workplace. A sound that is generated from within the inner ear is known as an otoacoustic emission (OAE). Distortion-product otoacoustic emissions (DPOAEs) assess evoked emission and hearing capacity. The aim of this study was to assess the signal-to-noise ratio in different frequencies and at different times of the shift work in workers exposed to various levels of noise. It was also aimed to provide a statistical model for signal-to-noise ratio (SNR) of OAEs in different frequencies based on the two variables of sound pressure level (SPL) and exposure time. MATERIALS AND METHODS: This case-control study was conducted on 45 workers during autumn 2014. The workers were divided into three groups based on the level of noise exposure. The SNR was measured in frequencies of 1000, 2000, 3000, 4000, and 6000 Hz in both ears, and in three different time intervals during the shift work. According to the inclusion criterion, SNR of 6 dB or greater was included in the study. The analysis was performed using repeated measurements of analysis of variance, spearman correlation coefficient, and paired samples t-test. RESULTS: The results showed that there was no statistically significant difference between the three exposed groups in terms of the mean values of SNR (P > 0.05). Only in signal pressure levels of 88 dBA with an interval time of 10:30-11:00 AM, there was a statistically significant difference between the right and left ears with the mean SNR values of 3000 frequency (P = 0.038). The SPL had a significant effect on the SNR in both the right and left ears (P = 0.023, P = 0.041). The effect of the duration of measurement on the SNR was statistically significant in both the right and left ears (P = 0.027, P < 0.001). CONCLUSION: The findings of this study demonstrated that after noise exposure during the shift, SNR of OAEs reduced from the beginning to the end of the shift.

Effects of Resveratrol on Crosstalk between Canonical Β-Catenin/Wnt and FOXO Pathways in Coronary Artery Disease Patients with Metabolic Syndrome: A Case Control Study.

Coronary artery disease (CAD) is the major cause of mortality and morbidity worldwide. The aim of this study was to explore the effect of resveratrol (RES) on Canonical ß-catenin/Wnt and forkhead box O (FOXO) pathways in CAD patients. We performed this study on 10 metabolic syndrome patients with three-vessel CAD and 10 sex-aged matched healthy subjects. The effects of RES on ß-Catenin, manganese superoxide dismutase (MnSOD), and peroxisome proliferator-activated receptor delta (PPAR-δ) expression were evaluated in peripheral blood mononuclear cells (PBMCs) of participants. RES could increase the MnSOD expression in CAD patients (38%, p < 0.0001). After RES treatment, the MnSOD expression of patients is still non-significantly lower than controls. In both blank and RES treatments, a significant positive correlation between ß-catenin and MnSOD mRNA expressions was found in controls, whereas no correlation between these gene expressions was found in untreated PBMCs of CAD patients. However, RES could modestly improve this pathway in CAD. RES could increase the MnSOD activity in healthy and CAD subjects (p = 0.051 and p = 0.009, respectively). Furthermore, in both blank and RES treatments, the significant correlation was found between total ß-catenin protein and the MnSOD activity in PBMCs of the controls but not in patients. The cross-talk between ß-catenin/Wnt and FOXO pathways was impaired in PBMCs of CAD patients. RES treatment could lead to a modest increase in the MnSOD activity independent of ß-catenin/FOXO pathway. Despite a modest improvement in the ß-catenin/FOXO pathway after RES treatment, this pathway was not completely repaired in CAD patients.

A Bioinformatics Approach to Prioritize Single Nucleotide Polymorphisms in TLRs Signaling Pathway Genes.

It has been suggested that single nucleotide polymorphisms (SNPs) in genes involved in Toll-like receptors (TLRs) pathway may exhibit broad effects on function of this network and might contribute to a range of human diseases. However, the extent to which these variations affect TLR signaling is not well understood. In this study, we adopted a bioinformatics approach to predict the consequences of SNPs in TLRs network. The consequences of non-synonymous coding SNPs (nsSNPs) were predicted by SIFT, PolyPhen, PANTHER, SNPs&GO, I-Mutant, ConSurf and NetSurf tools. Structural visualization of wild type and mutant protein was performed using the project HOPE and Swiss PDB viewer. The influence of 5'-UTR and 3'- UTR SNPs were analyzed by appropriate computational approaches. Nineteen nsSNPs in TLRs pathway genes were found to have deleterious consequences as predicted by the combination of different algorithms. Moreover, our results suggested that SNPs located at UTRs of TLRs pathway genes may potentially influence binding of transcription factors or microRNAs. By applying a pathway-based bioinformatics analysis of genetic variations, we provided a prioritized list of potentially deleterious variants. These findings may facilitate the selection of proper variants for future functional and/or association studies.

Evaluation of the effects of occupational noise exposure on serum aldosterone and potassium among industrial workers.

The existing literature indicates that occupational exposure to noise may have adverse effects on workers' health. The aim of this study was to evaluate the possible effects of exposure to different sound pressure levels (SPLs) on serum aldosterone and potassium concentration among Iranian blue collar workers in Golgohar Mining and Industrial Company in Sirjan, Kerman Province, Iran. This case-control study was performed on 45 workers of Golgohar Mining and Industrial Company. The subjects consisted of 30 workers from manufacturing departments and 15 office employees of the mining company. The controls, mainly with administrative jobs were exposed to 72 dBA SPL. Cases, in two separate groups, were exposed to noise levels of 88 dBA and 103 dBA, respectively. Noise intensity was measured at the desired locations. Noise measurements were performed according to the International Organization for Standardization (ISO) 9612. To measure the serum aldosterone and potassium concentrations, a 5 mL blood sample was taken from each worker at the specified time intervals and aldosterone concentration was determined using enzyme-linked immunosorbent assay (ELISA) test in the laboratory. Repeated measurement and Spearman's correlation coefficient analysis were used with α = 0.05. Exposure to the different levels of sound pressure resulted in different aldosterone concentrations and meanwhile an increase in the SPL did not affect the concentration of potassium. From 10:00 AM to 10:30 AM, as SPL increased, aldosterone concentrations did not increase significantly but from 13:30 PM to 14:00 PM, raised SPL led to a significant increase in aldosterone concentration. However, there was no correlation between the concentration of potassium and different factors. This study indicated that increases in SPLs affect aldosterone concentration but at the same time do not have significant effects on serum potassium level.

Association of miR-146a rs2910164 and miR-149 rs2292832 Variants with Susceptibility to Type 2 Diabetes.

BACKGROUND: The deregulation of miRNAs has been implicated in the pathogenesis of type 2 diabetes (T2D). Single nucleotide polymorphisms (SNPs) located within the miRNA sequence could alter miRNA maturation and expression or change the binding affinity of miRNAs to their target mRNAs. In the present study we aimed to elucidate the possible association between the miR-146a rs2910164 and miR-149 rs2292832 variants with the susceptibility to T2D and its related metabolic traits in an Iranian population. METHODS: The study population consisted of 183 type 2 diabetic and 192 non-diabetic subjects. The genotyping of the variants was performed by a PCR-RFLP method. RESULTS: The frequency of the CC genotype of the miR-146a rs2910164 variant was significantly higher in diabetic patients than in controls (15.85% vs. 7.81%, p = 0.043). The results of binary logistic regression suggested that this genotype was significantly associated with T2D (OR of 2.43 (95% CI 1.17 - 5.02, p = 0.016). Moreover, subjects carrying the CC genotype had significantly higher values for diastolic blood pressure, triglycerides, total cholesterol, fasting blood glucose and HbAlc levels compared to individuals having the GG and GC genotypes. Our bioinformatic analyses also showed that the miR-146a sequence is conserved across primate taxa and substituting G to C causes the structural instability of pre-miR-146a by changing the minimum free energy. For the rs2292832 variant, no statistically significant difference was detected for allele or genotype frequencies between T2D and control groups. CONCLUSIONS: Our findings suggest that miR-146a rs2910164 polymorphism might be associated with T2D and its cardiovascular risk factors in an Iranian population.

Evaluation of Distortion Product Otoacoustic Emissions (DPOAEs) among workers at an Industrial Company exposed to different industrial noise levels in 2014.

BACKGROUND: Noise-induced hearing loss (NIHL) is usually one of the main problems in industrial settings. The aim of this study was to determine whether changes in the signal-to-noise ratio (SNR) in different DPOAE are caused by exposure to different levels of noise at different time intervals among workers exposed to noise. METHODS: This case-control study was conducted in the autumn of 2014 on 45 workers at Gol Gohar Mining and Industrial Company, which is located in Sirjan in southeast Iran. The workers were divided into three groups based on their noise exposure, i.e., 1) 15 office workers as a control group with exposure to low levels of noise, 2) 15 workers from manufacturing departments who were exposed to a medium level of noise, and 3) 15 workers from manufacturing departments who were exposed to high levels of noise. The SNRs at the frequencies of 1000, 2000, 3000, 4000, and 6000 Hz were measured in both ears at three different time intervals during the shift work. SNRs of 6 or greater were considered as inclusion criterion. Repeated measures, the Spearman rank-order correlation test, and paired t-test analyses were used with α = 0.05 being the level of significance. RESULTS: For all frequencies in the right and left ears, the SNR values were more than 6, thus all SNR values were considered as acceptable responses. The effects of time and sound pressure level (SPL) on SNR were significant for the right and left ears (p = 0.027 and < 0.001, respectively). There was a statistically significant correlation between the SNR values in the right and left ears for the time intervals 7:30-8:00 A.M. and 13:30-14:00 P.M., which implied that an increase in the duration of exposure led to reduced SNR values (p = 0.024, r = 0.948). CONCLUSIONS: The comparison of the SNR values in the right and left ears (for all frequencies and the three different SPLs) indicated that the values decreased during the shift work.

Genetic Variants of Cytochrome b-245, Alpha Polypeptide Gene and Premature Acute Myocardial Infarction Risk in an Iranian Population.

BACKGROUND: Oxidative stress induced by superoxide anion plays critical roles in the pathogenesis of coronary artery disease (CAD) and hence acute myocardial infarction (AMI). The major source of superoxide production in vascular smooth muscle and endothelial cells is the NADPH oxidase complex. An essential component of this complex is p22phox, that is encoded by the cytochrome b-245, alpha polypeptide (CYBA) gene. The aim of this study was to investigate the association of CYBA variants (rs1049255 and rs4673) and premature acute myocardial infarction risk in an Iranian population. METHODS: The study population consisted of 158 patients under the age of 50 years, with a diagnosis of premature AMI, and 168 age-matched controls with normal coronary angiograms. Genotyping of the polymorphisms was performed by the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). RESULTS: There was no association between the genotypes and allele frequencies of rs4673 polymorphism and premature acute myocardial infarction (P>0.05). A significant statistical association was observed between the genotypes distribution of rs1049255 polymorphism and AMI risk (P=0.037). Furthermore, the distribution of AA+AG/GG genotypes was found to be statistically significant between the two groups (P=0.011). CONCLUSIONS: Our findings indicated that rs1049255 but not rs4673 polymorphism is associated with premature AMI.

Melatonin improve the sperm quality in forced swimming test induced oxidative stress in nandrolone treated Wistar rats.

This study investigates the effects of melatonin on the sperm quality and testis weight after the combination of swimming exercise and nandrolone decanoate (DECA). Two groups of male Wistar rats were treated for eight weeks as follows; group A consist of CO (control), Sham, N (DECA), S (swimming) and NS (DECA plus swimming); and group B: Sham M (sham melatonin), M (melatonin), MN (melatonin plus DECA), MS (melatonin plus swimming), MNS (melatonin, DECA plus swimming). The motility of sperm was significantly improved in melatonin groups in comparison to N, S and NS groups (P≤0.05).  The left testes weight was decreased in N, NS and MNS groups, and the right testes weight was decreased in N,S,NS, MS and MNS groups in compare with the control group. This study concluded that melatonin probably could improve the sperm motility and sex organs weight after the combination of DECA and exercise.

Triclabendazole (Anthelmintic Drug) Effects on the Excretory- Secretory Proteome of Fasciola hepatica in Two Dimension Electrophoresis Gel.

BACKGROUND: The aim of this study was to evaluate the protein spots of excretory - secretory products of Fasciola hepatica using two dimension electrophoresis method in the presence and absence of triclabendazole drug which can be considered to detect the target protein of the drug. METHODS: F. hepatica parasites were collected from infected cattle livers, divided in two groups and cultivated in RPMI 1640 medium. First group was treated with triclabendazole (TCBZ) and second group considered as control. The excretory-secretory (ES) products of each group were separated and total protein determined by Bradford method. To provide proteome spots, the ES proteins were precipitated and two dimension electrophoresis (2-DE) gel prepared. Protein amounts of two groups were compared using the statistical t-test and protein spots from 2-DE in test and control groups were also statistically analyzed. The protein spots of gels were identified by using protein database. RESULTS: The t-test showed a significant increase of total proteins in treated group (P<0.5). The protein spots count in the control group was less than test group however statistically not significant (p>0.05). Cathepsin L- protein (MW 36.7 pH 5.34), 14-3-3 epsilon 2 isoform (MW 28.2 pH 5.36), Cathepsin L1D (MW 36.5 pH 5.8) and Cathepsin L1D (MW 36.6 pH 6.26) were identified in test group. CONCLUSION: It seems that, these results can be considered to determine the proteins which the drug acts as a target on them.

Homocysteine induces heme oxygenase-1 expression via transcription factor Nrf2 activation in HepG2 cell.

BACKGROUND: Elevated level of plasma homocysteine has been related to various diseases. Patients with hyperhomocysteinemia can develop hepatic steatosis and fibrosis. We hypothesized that oxidative stress induced by homocysteine might play an important role in pathogenesis of liver injury. Also, the cellular response designed to combat oxidative stress is primarily controlled by the transcription factor Nrf2, a principal inducer of anti-oxidant and phase II-related genes. METHODS: HepG2 cells were treated with homocysteine in different time periods. Glutathione content was measured by flowcytometry. Using electrophoretic mobility shift assay (EMSA) and Western-blotting, anti-oxidant response element (ARE)-binding activity of Nrf2 for heme ocygenase-1 (HO-1) was demonstrated. Real time RT-PCR and Western-blotting were performed to evaluate whether homocysteine was able to induce mRNA and protein expression of HO-1. RESULTS: The role of Nrf2 in cellular response to homocysteine is substantiated by the following observations in HepG2 cells exposed to homocysteine (i) Western-blotting revealed that Nrf2 is strongly stabilized and became detectable in nuclear extracts. (ii) EMSA demonstrated increased binding of Nrf2 to oligomers containing HO-1 promoter-specific ARE-binding site. (iii) Real time RT-PCR and Western-blotting revealed increased mRNA and protein expression of inducible gene HO-1 after treatment with homocysteine. CONCLUSION: Data presented in the current study provide direct evidence that the immediate cellular response to oxidative stress provoked by homocysteine is orchestrated mainly by the Nrf2-ARE pathway. Therefore, induction of Nrf2-ARE-dependent expression of HO-1 could be a therapeutic option for hepatic cells damage induced by homocysteine.

Cytotoxicity of subtoxic AgNP in human hepatoma cell line (HepG2) after long-term exposure.

UNLABELLED: We aimed at evaluating the toxicity effects of low (subtoxic) concentrations of silver nanoparticles nanoparticles (AgNP, 5-10 nm) in human hepatoblastoma (HepG2) cell line after and during a period of about one month. METHODS: XTT and MTT assays were used to draw a dose-response curve; IC50 (half maximal inhibitory concentration) value of the AgNP on HepG2 cells was calculated to be 2.75-3.0 mg/l. The cells were exposed to concentrations of 0% (control), 1%, 4% and 8% IC50 of AgNP (corresponding to 0.00, 0.03, 0.12 and 0.24 mg/l of AgNP, respectively) for four consecutive passages. The treated cells were compared to the control group with respect to morphology and proliferation at the end of the period. RESULTS: The biochemical studies revealed significant increases of lactate dehydrogenase and alanine aminotransferase enzyme activity in the culture media of cells receiving 4% and 8% IC50; the increases in the aspartate aminotransferase enzyme activity and nitric oxide concentration became significant at 8% IC50. In the cell extracts, the average total protein and activity of glutathione peroxidase enzyme remained unchanged; the decrease in the average content of glutathione (GSH) and superoxide dismutase (SOD) activity became significant at 4% and 8% IC50. There were increases in lipid peroxidation (significant at 4% and 8% IC50) and cytochrome c content (significant at 8% IC50). The accumulations of the effects, during the experiment from one generation to the next, were not statistically remarkable except in cases of GSH and SOD. The results indicate clearly the involvement of oxidative changes in the cells after exposure to low doses of AgNP. CONCLUSION: The results might help specify a safer amount of AgNP for use in different applications.

The relationship between ACE gene insertion/deletion polymorphism and diabetic retinopathy in Iranian patients with type 2 diabetes.

BACKGROUND: The role of genetic factors in diabetic retinopathy (DR) is unclear. We investigated the relationship between DR and an insertion/deletion polymorphism in the angiotensin-converting enzyme (ACE) gene in Iranian patients with type 2 diabetes without overt nephropathy. METHODS: A total of 178 consecutive type 2 diabetic patients with DR (Group A) and 206 type 2 diabetic patients without DR (Group B) were studied. The following variables were determined: age, sex, body mass index, diabetes duration, medications used, history of coronary artery disease and its complications, blood pressure (systolic and diastolic), fasting plasma glucose, hemoglobin A1c, total cholesterol, low- and high-density lipoproteins, triglycerides, plasma creatinine, and 24-h urine albumin excretion. RESULTS: The groups were statistically similar in all variables except diabetes duration (P = 0.037), ACE activity (P < 0.001), and ACE genotype (P = 0.008). The DD genotype was significantly more common in Group A (32.6% versus 19.2% in Group B; P = 0.009). In multivariate regression analysis, the ID genotype (compared to the II genotype) was an independent predictor of DR (OR = 1.831, 95% CI = 1.074-3.124; P = 0.026). CONCLUSIONS: The D allele of the ACE gene is independently associated with DR in Iranian type 2 diabetic patients.

Polymorphism of Pro12Ala in the peroxisome proliferator-activated receptor gamma2 gene in Iranian diabetic and obese subjects.

BACKGROUND: Peroxisome proliferator-activated receptor gamma2 (PPARgamma2) is a nuclear receptor that regulates adipocyte differentiation, lipid metabolism, and insulin sensitivity. The aim of this study was to investigate the association between the Pro12Ala single nucleotide polymorphism (SNP) at the PPARgamma2 gene and type II diabetes (T2DM) and obesity in an Iranian population. METHODS: The genomic DNA of the 312 subjects included four groups: (1) nonobese with type II diabetes, (2) obese without type II diabetes, (3) obese with type II diabetes, and (4) nondiabetic nonobese controls. The Pro12Ala polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: Frequencies of the Ala allele in obese subjects were significantly different from those control subjects (odds ratio [OR], 2.358; 95% confidence interval [CI], 1.101-5.05) (P = 0.025). In contrast, no significant association was detected between the Pro12Ala polymorphism and type II diabetes (OR, 0.652; 95% CI, 0.261-1.628). In all subjects, the Ala carriers had a higher body mass index (BMI) compared with the common allele. CONCLUSIONS: Our results showed that the Pro12Ala polymorphism in the PPARgamma2 gene is associated with obesity in Iranian subjects and the presence of the Ala allele could predict higher BMI.