Biomarker-based validation of a food frequency questionnaire for the assessment of omega-3 fatty acid status in a healthy Iranian population.

There is no valid instrument to assess n-3 polyunsaturated fatty acids (n-3 PUFAs) intake in Iran. This study aims to develop a food frequency questionnaire (FFQ) that estimates the intake of n-3 PUFA and validate it in a healthy Iranian population based on the n-3 PUFA content of red blood cells (RBCs) and a 3-day food record (FR). A healthy population (n = 221) was recruited between February and July 2021. Participants completed the new FFQ and 3-day FR to evaluate the average intake of n-3 PUFAs. We used gas chromatography to assess the n-3 PUFA content of RBCs. To validate the FFQ based on FR and biomarker as references, the correlation coefficient was calculated. According to the Bland-Altman plots, a good agreement was found between the new FFQ and FR. Moreover, absolute intake values of ALA, EPA, DPA, DHA, and total n-3 PUFAs based on FFQ were positively correlated to their respective RBC membrane levels (coefficients between 0.205 and 0.508, p < 0.005) and FR (coefficients between 0.771 and 0.827, p < 0.001). This new FFQ is a valid instrument that can be applied to estimate the n-3 PUFA status of healthy Iranian adults.

Betanin alleviates oxidative stress through the Nrf2 signaling pathway in the liver of STZ-induced diabetic rats.

BACKGROUND: Continuing hyperglycemia causes and exacerbate oxidative stress. Betanin as the principal pigment of red beet root has antioxidant, anti-inflammatory, and anti-diabetic properties. The purpose of this study was to investigate the potency of betanin on antioxidant defense in STZ-induced diabetic rats' livers. METHODS: STZ at a single dose of 60 mg/kg body weight was intraperitoneally injected and betanin (10, 20, and 40 mg/kg/day) was administered orally for 28 days. Malondialdehyde (MDA), total antioxidant capacity (TAC), protein carbonyl (PC) levels, and the enzyme activity of superoxide dismutase (SOD), catalases and glutathione peroxidases (GPx) were evaluated in the liver. Furthermore, gene expression of Nrf2 and mentioned antioxidant enzymes were measured by Real-time PCR. RESULTS: Betanin (10 and 20 mg/kg) significantly reduced PC levels and increased antioxidant enzyme activity in diabetic rats compared to the control diabetic group (P < 0.01). In comparison to the diabetic control group, all studied genes expression in diabetic rats were increased significantly with betanin at doses of 10 and 20 mg/kg (P < 0.02). The increase in gene expression at 20 mg/kg of betanin was significantly stronger than others (P < 0.015) except for the catalase (P = 0.201), that was almost the same. Moreover, treatment of diabetic rats with 20 mg/kg of betanin could significantly increase TAC levels (P < 0.05) and decrease MDA levels (P < 0.001) compared to diabetic control group. CONCLUSIONS: Betanin could increase the antioxidant capacity of liver tissue associated with the Nrf2-mediated pathway in a dose-dependent manner.

The anti-diabetic effects of betanin in streptozotocin-induced diabetic rats through modulating AMPK/SIRT1/NF-κB signaling pathway.

BACKGROUND: In the last few years, the effects of bioactive food components have received much attention because of their beneficial effects including decreasing inflammation, scavenging free radicals, and regulating cell signaling pathways. Betanin as a potent antioxidant has been previously reported to exhibit anti diabetic effects. The present study aimed to evaluate the effects of betanin on glycemic control, lipid profile, hepatic function tests, as well as the gene expression levels of 5' adenosine monophosphate­activated protein kinase (AMPK), sirtuin-1 (SIRT1), and nuclear factor kappa B (NF­κB) in streptozocin (STZ) induced diabetic rats. METHODS: Diabetes was induced in male Sprague-Dawley rats by intraperitoneal administration of STZ. Different doses of betanin (10, 20 and 40 mg/kg.b.w) was administered to diabetic rats for 28 days. Fasting blood glucose and serum insulin were measured. The histopathology of liver and pancreas tissue evaluated. Real-time PCR was performed to assess gene expression levels. RESULTS: Treatment of diabetic rats with betanin (10 and 20 mg/kg.b.w) reduced FBG levels compared to the control diabetic rats (P < 0.001). Betanin at the dose of 20 mg/kg.b.w was most effective in increasing serum insulin levels (P < 0.001) improving glucose tolerance test (GTT) as well as improvement in lipid profile and liver enzymes levels. According to histopathologic assay, different damages induced by STZ to liver and pancreas tissues was largely eliminated by treatment with 10 and 20 mg/kg.b.w of betanin. Betanin also significantly upregulated the AMPK and SIRT1 and downregulated the NF-κB mRNA expression compared to the diabetic control rats (P < 0.05). CONCLUSION: Betanin could modulate AMPK/SIRT1/NF-κB signaling pathway and this may be one of its anti-diabetic molecular mechanisms.

Ferritin Degradation by Pneumococcal HtrA, RadA and ClpP Serine Proteases : A Probable Way For Releasing and Acquisition Of Iron.

PURPOSE: Iron is a necessary element for the growth of bacteria; however, there are limited iron sources known for these microorganisms yet. Intracellular iron is stored as ferritin from, which releases iron in a gradual and controlled manner. The present study aimed to characterize ferritin-binding proteins (FBPs) of Streptococcus pneumoniae. MATERIAL AND METHODS: S. pneumoniae species were cultured in BHI broth containing ferritin (1094 ng/mL) for 4h at 37°C. Ferritin level was measured using ELISA assay. Bacterial proteome was electrophoresed on SDS-PAGE and then transferred on PVDF nitrocellulose membrane. Afterward, the PVDF membranes were incubated with a ferritin solution. Identification of ferritin binding proteins was performed using anti-ferritin monoclonal antibody conjugated with HRP enzyme. Molecular docking was used to assess the interaction between pneumococcal proteases and FBPs applying phenylmethylsulfonyl fluoride (PMSF) as a protease inhibitor. RESULTS: No FBPs were identified in S. pneumoniae proteome. Moreover, ferritin levels have significantly (p<0.05) decreased following the growth of S. pneumoniae in ferritin-rich BHI medium. Also, molecular docking showed that RadA protease, ClpP hydrolase, and HtrA protease can potentially interact with PMSF protease inhibitors. On the other hand, the addition of the PMSF to the culture of S. pneumoniae prevented the reduction of ferritin, which indicates a potential role of RadA, ClpP, and HtrA proteases in ferritin degradation. CONCLUSION: Our results suggest that S. pneumoniae produces no FBPs and also cannot directly use ferritin as an iron source. However, ferritin may be degraded through a protease-mediated mode.

Effects of crocin and saffron aqueous extract on gene expression of SIRT1, AMPK, LOX1, NF-κB, and MCP-1 in patients with coronary artery disease: A randomized placebo-controlled clinical trial.

This trial evaluated the potential impacts of saffron aqueous extract (SAE) and its main carotenoid on some of the atherosclerosis-related gene expression and serum levels of oxidized low-density cholesterol (ox-LDL) and Monocyte chemoattractant protein 1 (MCP-1) in patients with coronary artery disease (CAD). Participants of this randomized controlled trial included 84 CAD patients who categorized into three groups: Group 1 received crocin (30 mg/day), Group 2 SAE (30 mg/day), and Group 3 placebo for 8 weeks. Gene expression of Sirtuin 1 (SIRT1), 5'-adenosine monophosphate-activated protein kinase (AMPK), Lectin-like oxidized LDL receptor 1 (LOX1), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and MCP-1 in peripheral blood mononuclear cells assessed by real-time PCR. Furthermore, serum ox-LDL and MCP-1 levels measured at the beginning and end of the intervention. Compared with the placebo group, gene expression of SIRT1 and AMPK increased significantly in the crocin group (p = .001), and the expression of LOX1 and NF-κB decreased significantly (p = .016 and .004, respectively). Serum ox-LDL levels decreased significantly in the crocin group after the intervention (p = .002) while MCP-1 levels decreased both in crocin and SAE groups (p = .001). Crocin may have beneficial effects on CAD patients by increasing the gene expression of SIRT1 and AMPK and decreasing the expression of LOX1 and NF-κB.

Saffron and crocin improved appetite, dietary intakes and body composition in patients with coronary artery disease.

Introduction: Central obesity is an independent risk factor for coronary artery disease (CAD). It can increase cardio-metabolic risks through hypertension, hyperlipidemia and insulin resistance. Saffron and its bioactive compounds (crocin and crocetin) can modify some of metabolic disorders through multiple mechanisms. The aim of this study was to assess the efficacy of saffron and crocin on lipid profile, appetite, dietary intakes, anthropometric indices and body composition in patients with CAD. Methods: This 8 weeks randomized, double-blind, and placebo-controlled trial was conducted on 84 patients with CAD between the ages of 40 and 65 years old. Participants were randomly divided into groups to receive a daily supplement of 30 mg saffron aqueous extract (SAE) or 30 mg crocin or placebo. Appetite, dietary intake, anthropometry, body composition, biochemical analysis were assessed before and after the study. Results: In SAE and crocin group, anthropometric and some body composition variables revealed a pattern of improvement after intervention. Decrease in body mass index (BMI), waist circumference and fat mass values in SAE group was significantly more than crocin group (P < 0.001). There was no significant difference at the end of study in lipid profile parameters. Both SAE and crocin yielded significant decrease in energy and dietary intake mean values (P < 0.001 and P = 0.046), while it remained unchanged in the placebo group, also the appetite decreased significantly in SAE and crocin group (P < 0.001 and P = 0.029, respectively). Conclusion: The results of present study regarding anti-obesity feature of SAE and crocin in patients with CAD was promising. However the SAE was better in appetite suppressing, dietary intake and central obesity reduction.

Allelic Frequency of a 24-bp Duplication in Exon 10 of the CHIT1 Gene in the General Iranian Population.

BACKGROUND: The human chitinase chitotriosidase enzyme, which is encoded by the CHIT1 gene, is produced by macrophages, and may be important in immune responses to chitin-containing organisms, such as fungi. Plasma chitotriosidase activity is used to diagnose and monitor some forms of lysosomal storage disorders, such as Gaucher's and Niemann-Pick disease. However, homozygous duplication of a 24-bp region in exon 10 of the CHIT1 gene eliminates enzyme activity and may complicate disease monitoring. The high prevalence of this mutation highlights the need to determine its frequency in different populations and screen patients for this mutation to verify whether chitotriosidase activity is a reliable marker of lysosomal storage disease. This study investigated the allele frequency of the 24-bp duplication in the general Iranian population. METHODS: To identify the 24-bp duplication in exon 10 of the CHIT1 gene (H allele), genotyping of DNA extracted from peripheral blood leukocytes of 577 healthy Iranians was performed using polymerase chain reaction (PCR) amplification and high resolution melting (HRM) PCR techniques. RESULTS: In this study, heterozygous and homozygous duplications were detected in 183 (31.7%) and 35 (6.1%) subjects, respectively. In addition, the allelic frequency was 21.9% (95% confidence interval). CONCLUSION: Our study indicates that genotype analysis by HRM-PCR is a fast, reliable, and highly accurate screening approach for identifying the 24-bp duplication in CHIT1 exon 10. Due to the wide range of duplication frequencies among different ethnic groups, new biomarkers are necessary for assessing genetic characteristics of lysosomal storage disorders in different populations.

In vitro effect of lead, silver, tin, mercury, indium and bismuth on human sperm creatine kinase activity: a presumable mechanism for men infertility.

BACKGROUND: The aim of the present study was to investigate the in vitro effects of mercury (Hg+2), lead (Pb+2), silver (Ag+2), tin (Sn+2), bismuth (Bi+3) and indium (In+3) ions on sperm creatine kinase. METHODS: creatine kinase was isolated from human sperm homogenates after chromatography on a DEAE cellulose column. RESULTS: At 60 mug ml-1 metal concentration, 70% of the creatine kinase activity was inhibited by Hg+2, while at the same concentration, Pb+2, Ag+2, Sn+2, Bi+3 and In+3 caused 68%, 66.5%, 65.7%, 64.7% and 62.7% inhibition, respectively. All six metal ions displayed a competitive type of inhibition mechanism for the isolated creatine kinase as analyzed by Lineweaver-Burk plot. Ki values of Hg+2, Pb+2, Ag+2, Sn+2, Bi+3 and In+3 were calculated and 8.34 mM, 5 mM, 4.54 mM, 3.45 mM, 3.12 mM and 2.63 mM values were obtained, respectively. CONCLUSION: All the studied metal ions, at levels of 60 mug ml-1, may reduce normal sperm metabolism by inhibition of sperm creatine kinase, which probably is an important cause of infertility in men. However, further investigations, as in vitro and in vivo, are needed to elucidate the exact mechanism of heavy metals on male reproductive functioning at the molecular level.