The Effects of Synbiotic Supplementation on Carotid Intima-Media Thickness,Biomarkers of Inflammation,and Oxidative Stress in People with Overweight,Diabetes, and Coronary Heart Disease: a Randomized,Double-Blind,Placebo-Controlled Trial

Synbiotics are known to exert multiple beneficial effects, including anti-inflammatory and antioxidant actions. The aim of this study was to evaluate the effects of synbiotic supplementation on carotid intima-media thickness (CIMT), biomarkers of inflammation, and oxidative stress in people with overweight, diabetes, and coronary heart disease (CHD). This randomized, double-blind, placebo-controlled trial was conducted and involved 60 people with overweight, diabetes, and CHD, aged 50–85 years old. Participants were randomly allocated into two groups to take either synbiotic supplements containing three probiotic bacteria spices Lactobacillus acidophilus strain T16 (IBRC-M10785), Lactobacillus casei strain T2 (IBRC-M10783), and Bifidobacterium bifidum strain T1 (IBRC-M10771) (2 × 10⁹ CFU/g each) plus 800 mg inulin or placebo (n = 30 each group) for 12 weeks. Fasting blood samples were taken at baseline and after the 12-week intervention period to determine metabolic variables. After the 12-week intervention, compared with the placebo, synbiotic supplementation significantly reduced serum high-sensitivity C-reactive protein (hs-CRP) (− 3101.7 ± 5109.1 vs. − 6.2 ± 3163.6 ng/mL, P = 0.02), plasma malondialdehyde (MDA) (− 0.6 ± 1.0 vs. − 0.1 ± 0.3 μmol/L, P = 0.01), and significantly increased nitric oxide (NO) levels (+ 7.8 ± 10.3 vs. − 3.6 ± 6.9 μmol/L, P < 0.001). We did not observe any significant changes of synbiotic supplementation on other biomarkers of oxidative stress and CIMT levels. Overall, synbiotic supplementation for 12 weeks among people with overweight, diabetes, and CHD had beneficial effects on serum hs-CRP, plasma NO, and MDA levels; however, it did not have any effect on other biomarkers of oxidative stress and CIMT levels.

     

The methylation status of TNF-α and SOCS3 promoters and the regulation of these gene expressions in patients with Behçet’s disease

Abstract

Introduction: The aim of this study was to evaluate the methylation status of TNF-α and SOCS3 promoters in patients with BD and compare them with a healthy group.

Method: This was a case–control study, in which 47 subjects with BD and 61 individuals as the control participated. Blood samples were collected from all the participants. Then, PBMCs were isolated using the Ficoll method and methylation of considered sites was investigated using the qMS-PCR technique after DNA extraction by the rapid genomic DNA extraction method and its analysis with Nano-drop.

Results: The methylation and expression of TNF-α showed that the methylation level significantly declined in the patient in comparison with the healthy (p?<?0.05). Moreover, the results on the mean expression showed that it significantly increased in the patient group, as compared with the healthy group (p?<?0.05). In addition, the expression of the SOCS3 gene was not significantly different between the patients and healthy subjects while the level of SOCS3 methylation was significantly higher in the patient group than that in the healthy group (p?<?0.05).

Discussion: The present study revealed that the gene expression of TNF-alpha increased in BD patients, suggesting that TNF-alpha likely has a role in the pathogenesis of BD.     

Association of interleukin‐17 gene polymorphisms and susceptibility to brucellosis in Hamadan,western Iran

IL‐17is one of the most important inflammatory cytokines that stimulate immunity responses in humans infected with Brucella species, acting as a regulator that reduces release of γ‐IFN, thus increasing resistance to brucellosis. Gene polymorphisms in the regulatory regions of cytokine‐encoding genes affect the amountsof cytokines produced and play a fundamental role in infectious diseases. The aim of this study was to determine the association between IL‐17 gene polymorphisms and susceptibility to brucellosis. In this case‐control study, 86 patients with brucellosis and 86 healthy persons in Hamadan, western Iran, from September 2014 to September 2016, were included. IL‐17 genetic variants at positions rs4711998 A/G, rs8193036 C/T, rs3819024 A/G, rs2275913 A/G, rs3819025 A/G, rs8193038 A/G, rs3804513 A/T, rs1974226 A/G and rs3748067 A/G were analyzed by restriction fragment length polymorphism‐PCR. Serum IL‐17 titers were measured by sandwich ELISA. GG genotypes at positions rs4711998 and rs3748067 were present significantly more frequently in patients with brucellosis than in controls (P < 0.05). The AA genotype at positions rs4711998, rs2275913 and rs3748067 and GG genotype at position rs19744226 were present significantly more frequently in controls than in the patient group. These results suggest that the AA genotype at positions rs3748067, rs3819025 and rs4711998 and GG genotype at position rs3819024 are likely protective factors against brucellosis, whereas the GG genotype at positions rs3748067, rs3819025 and rs4711998 and AA genotype at position rs3819024 may be risk factors against the disease. No significant relationships were found between serum IL‐17 titers and genotypes of the single‐nucleotide polymorphisms.     

Fatty Acid Patterns of Seeds of Some Salvia Species from Iran – A Chemotaxonomic Approach

In this study, the seed oil content and fatty acid (FA) profile of 21 populations from 16 wild Salvia species of Iran were analyzed by GC. Patterns of chemical variations of the oils among species were identified via numerical analyses and also the taxonomic status of the infrageneric grouping was outlined in the genus. Salvia species were scored based on the contents of main FAs using principal coordinate analysis (PCO). The results showed that the total oil content in the seeds varied significantly, and ranged from 6.68 to 38.53% dry weight. α‐Linolenic (18:3ω3, 1.69 – 53.56%), linoleic (18:2ω6, 13.04 – 60.64%), oleic (18:1ω9, 6.15 – 27.06%), palmitic (16:0, 3.77 – 9.27%), and stearic (18:0, 1.78 – 3.05%) acid were identified as five major FAs in the oils. The amount of ω‐3 and ω‐6 FAs ranged between 1.90 – 53.80% and 13.46 – 60.83% of total FAs in the seed oils, respectively. The results confirmed that FA profiles were distinctive among the species and that they can be used as chemotaxonomic markers. The discrimination of Salvia species according to their botanical classification at intersectional level was supported. In general, seed oils of Salvia species were rich sources of polyunsaturated FAs, except in linoleic and α‐linolenic acid, and may be valuable for food and pharmaceutical industries.     

Interactions of anthropometric indices,rs9939609 FTO gene polymorphism and breast cancer: A case-control study

Contradictory results were reported on the effect of fat mass- and obesity-associated (FTO) gene and anthropometric measurements on breast cancer (BC). This study aimed to assess the interactions between rs9939609 polymorphism of FTO gene, anthropometric indices and BC risk in Iranian women. This case-control study was performed on 540 women including 180 women with BC and 360 healthy women in Tehran, Iran. Physical activity and dietary intakes were assessed by validated questionnaires. Data on sociodemographic and pathologic factors of the participants as well as their blood samples were collected. The rs9939609 FTO gene polymorphism was genotyped using the tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR). No significant association was found between BC and risk allele of FTO rs9939609 polymorphism after adjustments for the confounders. However, there was a significant association between rs9939609 polymorphism risk allele and BC risk in females with overweight, even after adjusting for age, family history of BC, abortion, BMI and the number of pregnancies (P < .05). The association was disappeared after further adjustments for lifestyle factors including smoking, alcohol consumption, calorie and macronutrients intake, and physical activity. The FTO gene polymorphism was associated with the risk of BC in overweight individuals. This association was influenced by environmental factors including diet, alcohol consumption and smoking. Future studies are required to confirm the association between the FTO gene and BC in overweight females and to identify the underlying mechanisms.     

Successful seed germination of the nickel hyperaccumulator Stackhousia tryonii

* BACKGROUND AND AIMS: Stackhousia tryonii, a rare nickel hyperaccumulating herb, is endemic to ultramafic (serpentine) soils of central Queensland, Australia. The effects of eight dormancy-relieving treatments on germination of stored seeds of Stackhousia tryonii were investigated under controlled light and temperature conditions. * METHODS: The treatments were: untreated (control i), leached and dehydrated (primed control ii), treating with gibberellic acid (150 and 300 microM), smoke extract (5 and 10 %, v/v) and potassium cyanide (40 and 80 mM). * KEY RESULTS: Freshly harvested seeds did not germinate. Germination percentage increased with time of storage for up to 18 months (38.3 %). Gibberellin, smoke extract and cyanide treatments did not significantly improve germination. Light did not affect seed germination and there was no interaction between dormancy-relieving treatments and light. A significant inhibition of germination occurred in seeds treated with 5 % (but not 10 %) aqueous smoke extract. Saturated fatty acids, predominantly tridecanoic (C13:0), constituted about 90 % of the total fatty acids in the oil of freshly harvested seeds. In contrast, there was increased accumulation (>75 %) of mono-unsaturated (oleic, c18:1) and poly-unsaturated (linoleic, c18:2; linolenic, c18:3) fatty acids in the oil of stored seeds. * CONCLUSIONS: Seeds of S. tryonii require an after-ripening period for germination.     

Sulfhydryl-based tumor antigen-carrier protein conjugates stimulate superior antitumor immunity against B cell lymphomas

Therapeutic vaccination of B cell lymphoma patients with tumor-specific Ig (idiotype, or Id) chemically coupled to the immunogenic foreign carrier protein keyhole limpet hemocyanin (KLH) using glutaraldehyde has shown promising results in early clinical trials, and phase III trials are underway. However, glutaraldehyde Id-KLH vaccines fail to elicit anti-Id immune and clinical responses in many patients, possibly because glutaraldehyde reacts with lysine, cysteine, tyrosine, and histidine residues, damaging critical immunogenic epitopes. A sulfhydryl-based tumor Ag-carrier protein conjugation system using maleimide chemistry was used to enhance the efficacy of Id-KLH vaccines. Maleimide Id-KLH conjugates eradicated A20 lymphoma from most tumor-bearing mice, whereas glutaraldehyde Id-KLH had little efficacy. Maleimide Id-KLH elicited tumor-specific IgG Abs and T cells, with CD8(+) T cells being the major effectors of antilymphoma immunity. Maleimide Id-KLH vaccines also demonstrated superior efficacy in 38C13 and BCL-1 lymphoma models, where Abs were shown to be critical for protection. Importantly, standard glutaraldehyde Id-KLH conjugation procedures could result in "overconjugation" of the tumor Ag, leading to decreased efficacy, whereas the heterobifunctional maleimide-based conjugation yielded potent vaccine product regardless of conjugation duration. Under lysosomal processing conditions, the Id-carrier protein linkage was cleavable only after maleimide conjugation. Maleimide KLH conjugation was easily performed with human Igs analogous to those used in Id-KLH clinical trials. These data support the evaluation of sulfhydryl-based Id-KLH vaccines in lymphoma clinical trials and possibly the use of tumor Ag-carrier protein vaccines for other cancers.     

Chronic type-I diabetes could not impede the anti-inflammatory and anti-apoptotic effects of combined postconditioning with ischemia and cyclosporine A in myocardial reperfusion injury

It has been shown that diabetes modifies the myocardial responses to ischemia/reperfusion (I/R) and to cardioprotective agents. In this study, we aimed to investigate the effects of combined treatment with ischemic postconditioning (IPostC) and cyclosporine A (CsA) on inflammation and apoptosis of the diabetic myocardium injured by I/R. Eight weeks after induction of diabetes in Wistar rats, hearts were mounted on a Langendorff apparatus and were subsequently subjected to a 30-min regional ischemia followed by 45-min reperfusion. IPostC was induced at the onset of reperfusion, by 3 cycles of 30-s reperfusion/ischemia (R/I). The concentration of creatine kinase (CK), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were determined; the levels of total and phosphorylated glycogen synthase kinase 3 beta (p-GSK3β) and B-cell lymphoma 2 (Bcl-2) were quantified by western blotting, and the rate of apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. Administration of either IPostC or CsA alone in nondiabetic animals significantly reduced CK, TNF-α, IL-1β, and IL-6 concentrations, increased the p-GSK3β and Bcl-2, and decreased the level of apoptosis (P < 0.05) but had no effect on diabetic hearts. However, in diabetic animals, after administration of CsA, the cardioprotective effects of IPostC in increasing the p-GSK3β and Bcl-2 and decreasing apoptosis and inflammation were restored in comparison with nonpostconditioned diabetic hearts. IPostC or CsA failed to affect apoptosis and inflammation and failed to protect the diabetic myocardium against I/R injury. However, combined administration of IPostC and CsA at reperfusion can protect the diabetic myocardium by decreasing the inflammatory response and apoptosis.