Efficacy of high-intensity interval and continuous endurance trainings on cecal microbiota metabolites and inflammatory factors in diabetic rats induced by high-fat diet.

Physical exercise is known to modulate the intestinal microbiota composition and control the symptoms of metabolic syndrome. In this research, we intend to investigate and compare the effect of high-intensity interval and continuous endurance trainings (HIIT and CET) on cecal microbiota metabolites and inflammatory factors in diabetic rats. A number of Wistar rats were made diabetic by a high-fat diet and trained under two types of exercise protocols, HIIT and CET. After taking samples from the cecal tissue and serum of rats to reveal the effect of exercise, three microbial species from the Firmicute and Bacteroid phyla, which are the main types of intestinal microbes, and their metabolites include two short-chain fatty acids (SCFAs), butyrate and propionate and also, the inflammatory factors TLR4 and IL6 were analyzed through quantitative polymerase chain reaction (qPCR), high-performance liquid chromatography (HPLC), and Enzyme-linked immunosorbent assay (ELISA) methods. In general, exercise while increasing the representative of Firmicute has caused a relative reduction of Bacteroides and improved the concentration of SCFAs. In this regard, HIIT outperforms CET in up-regulating Akkermansia and Butyrivibrio expression, and butyrate and propionate metabolites concentration. Also, both exercises significantly reduced cecal expression of TLR4 and sera concentration of IL6 compared to the diabetic group, although the reduction rate was higher in the CET group than in HIIT. Our findings suggest that some symptoms of metabolic syndrome such as intestinal dysbiosis and the resulting metabolic disorders are better controlled by HIIT and inflammation by CET. Certainly, more extensive research on other contributing factors could help clarify the results.

Benzenesulfonamide as a novel, pharmaceutical small molecule inhibitor on Aß gene expression and oxidative stress in Alzheimer's Wistar rats.

Alzheimer's disease (AD) is the most prevalent acute neurodegenerative disease described by memory loss and other cognitive functions. Benzenesulfonamide, a novel, potent, and small organic molecule, was synthesized to investigate its effects on the levels of oxidative biomarkers (GPx, ROS, and MDA) and expression of beta-amyloid peptides (Aß40 and Aß42) in the pathology of AD. The results were compared with the rivastigmine drug. Applying benzenesulfonamide to Alzheimer's-induced Wistar rats showed a significant increase in the level of oxidative biomarkers (GPx, ROS, and MDA) in both the brain and blood serum as well as amyloid-ß40 and 42 gene expressions. Therefore, benzenesulfonamide could be considered a novel therapeutic agent against AD.

Dual-emissive phenylalanine dehydrogenase-templated gold nanoclusters as a new highly sensitive label-free ratiometric fluorescent probe: heavy metal ions and thiols measurement with live-cell imaging.

Phenylalanine dehydrogenase (PheDH) has been proposed as an ideal protein scaffold for the one-step and green synthesis of highly efficient multifunctional gold nanoclusters. The PheDH-stabilized fluorescent gold nanoclusters (PheDH-AuNCs) with dual emission/single excitation exhibited excellent and long-term stability, high water solubility, large Stokes shift and intense photoluminescence. Selectivity studies demonstrated that the red fluorescence emission intensity of PheDH-AuNCs was obviously decreased in less than 10 min by the addition of mercury, copper, cysteine or glutathione under the single excitation at 360 nm, without significant change in the blue emission of the PheDH-AuNCs. Therefore, the as-prepared PheDH-AuNCs as a new excellent fluorescent probe were successfully employed to develop a simple, rapid, low cost, label- and surface modification-free nanoplatform for the ultrasensitive and selective detection of Hg2+, Cu2+, Cys and GSH through a ratiometric fluorescence system with wide linear ranges and detection limits of 1.6, 2.4, 160 and 350 nM, respectively which were lower than previous reports. In addition, the results showed that PheDH-AuNCs can be used for the detection of toxic heavy metal ions and small biomarker thiols in biological and aqueous samples with acceptable recoveries. Interestingly, PheDH-AuNCs also displayed a promising potential for live-cell imaging due to their low toxicity and great chemical- and photo-stability.

Attachment and Emotion Regulation: A Cross-Cultural Comparative Study of Iranian and Dutch Gay Men.

Negative beliefs and stigmatization of all that is not heterosexual still have an adverse impact on the mental health of the homosexual population in many countries. The purpose of the present study was to comparatively assess how sociocultural differences regarding the level of homosexuality acceptance impact the adult attachment dimensions and emotion regulation strategies among three groups of Iranian and Dutch gay men. A community sample of 124 gay men (40 of Iranians residing in their home country, 41 of Iranians immigrated to the Netherlands and 43 Dutch) participated in the study and completed Revised Adult Attachment Scale (RAAS) and Emotion Regulation Questionnaire (ERQ). MANOVA and follow-up post-hoc Tukey tests were conducted in order to analyze the data. Results demonstrated a noticeable difference in both studied variables, attachment dimensions (close, anxiety, and depend) and emotion regulation strategies (cognitive reappraisal and expressive suppression), among the three groups of participants. As the Iranian (residing in Iran) group showed the greatest levels of anxiety and emotional suppression with the lowest levels of close (convenience of getting intimate to others), depend (trust and depend on others to be availabale when needed) and cognitive reappraisal (ability to alter the emotion caused by an event, before experiencing it by reinterpreting the situation), while the highest levels of depend, close and cognitive reappraisal and the lowest levels of anxiety and emotional suppression were seen in the Dutch group. Finally, Iranian gay immigrants came half way between. This data highlights the role of cultural differences in terms of homosexuality acceptance or stigmatization, in the way gay men exhibit their attachment and manage their emotions either by reappraisal or suppression. Comparative cross-cultural studies are possibly able to open paths to new research on psychological factors of non-heterosexuals in different countries with various cultures and religions.

Screening and Identification of DNA Nanostructure Aptamer Using the SELEX Method for Detection of Epsilon Toxin.

Background: Epsilon toxin (ETX), produced by Clostridium perfringens, is one of the most potent toxins known, with a lethal potency approaching that of botulinum neurotoxins. Epsilon toxin is responsible for enteritis. Therefore, the development of rapid and simple methods to detect ETX is imperative. Aptamers are single-stranded oligonucleotides that can bind tightly to specific target molecules with an affinity comparable to that of monoclonal antibodies (mAbs). DNA aptamers can serve as tools for the molecular identification of organisms, such as pathogen subspecies. Objectives: This study aimed to isolate high-affinity single-stranded DNA (ssDNA) aptamers against ETX. Methods: This study identified aptamers using the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method, enzyme-linked apta-sorbent assay (ELASA), and surface plasmon resonance (SPR) to determine the affinity and specificity of the newly obtained aptamers targeting ETX. Results: Several aptamers obtained through the SELEX process were studied. Among them, 2 aptamers, ETX clone 3 (ETX3; dissociation constant (Kd = 8.4 ± 2.4E-9M) and ETX11 (Kd = 6.3 ± 1.3E-9M) had favorable specificity for ETX. The limits of detection were 0.21 and 0.08 µg/mL for ETX3 and ETX11, respectively. . Conclusions: The discovered aptamers can be used in various aptamer-based rapid diagnostic tests for the detection of ETX.

The Serum Oxidative Stress Biomarkers and Selenium Levels in a Group of Migraine Patients Compared with Healthy Controls: a Case-Control Study.

Migraine is one of the most common neurological disorders associated with recurrent attacks of moderate to severe headache. Oxidative stress may play an important role in migraine pathogenesis. This study aimed to measure and compare the serum levels of Selenium, total antioxidant capacity (TAC), and malondialdehyde) MDA (in migraine patients and healthy individuals. This case-control study was performed on 31 migraine patients and 30 age and gender-matched healthy controls. The severity of headache was assessed with a standard questionnaire, and the serum levels of Selenium (Se), MDA, and TAC were measured via biochemical methods. The odds of migraine were calculated across quartile of Se and oxidative stress biomarkers via binary logistic regression. Migraine patients had a significant lower Se levels (81.06 ± 8.66 vs. 88.94 ± 10.23 µg/L, P = 0.002) and a significant higher MDA levels (3.04 ± 1.74 vs. 2.06 ± 0.59 nmol/ml, P = 0.005) compared to healthy participants. Although serum TAC levels (1.34 ± 0.34 vs.1.37 ± 0.33 mmol/L, P = 0.755) were not significantly different between migraine patients rather than healthy subjects. Individuals in the lowest quartile of Se levels were about eleven times more likely to have migraine than those in the highest quartile (OR: 11.2; 95%CI: 1.57 to 80.2; P-trend: 0.016). Besides, being in the highest quartile of the serum MDA level, the odds of having migraine increases 15.4 times compared to the lowest quartile (OR = 15.4, 95%CI: 1.1 to 221, P = 0.044). No significant association was found between TAC and migraine. The lower Se and MDA levels in migraine patients gives rise to the probability which oxidant status may play an underlying role in migraine pathophysiology.

The therapeutic effects of berberine plus sitagliptin in a rat model of fatty liver disease.

OBJECTIVES: Fatty liver disease (FLD) is a disorder related to accumulation of excess fat within the hepatocytes. In this study, the effects of Berberine, a natural compound, and Sitagliptin as a DPP-4 inhibitor, were observed in a rat model of FLD. MATERIALS AND METHODS: Forty male rats were divided into five groups (n=6) including the control group (normal food and water), high-fat group (high-fat diet (HF) for 6 weeks), Berberine group (HF with oral administration of Berberine at 150 mg/kg for 6 weeks), Sitagliptin group (HF with oral administration of Sitagliptin at 10 mg/kg for 6 weeks), and Berberine/ Sitagliptin group (HF diet within combination with oral administration of Berberine 75 mg/kg and Sitagliptin 5 mg/kg for 6 weeks). Animals were examined for weight gain, serum and hepatic biochemical parameters, tissue histology, expression of glucose transporter type 4 (GLUT4) mRNA, and protein expression of Adiponectin receptor2 (AdipoR2) and extracellular signal-regulated kinase (ERK) and phoERK. RESULTS: The results showed that ALT, AST, lipid profile, insulin, glucose, MDA, and TNF-α were significantly improved in high-fat rats treated with Berberine/ Sitagliptin compared with HF and Sitagliptin, and Berberine alone groups. SOD and adiponectin levels in Berberine/ Sitagliptin group were also significantly increased compared with the other groups. Immunoblot analysis showed that the expression of pho-ERK/ERK was significantly decreased and expression of AdipoR2 significantly increased in the Berberine/ Sitagliptin group compared with other groups. CONCLUSION: Co-administration of Berberine and Sitagliptin is an effective therapeutic regimen for conditions associated with hyperlipidemia.

Evaluation of the Effect of Vitamin D Supplementation on Anthropometric Indicators and Dietary Intake of Patients with Type 2 Diabetes.

BACKGROUND: Various studies have shown that diabetes and its complications are associated with vitamin D deficiency. Due to the possible role of vitamin D in reducing the complications of diabetes and the high prevalence of its deficiency in Iran, this study was designed to investigate the effect of vitamin D supplementation on anthropometric indices and dietary intake of patients with type 2 diabetes. METHODS: This randomized clinical trial (RCT) study was performed on 74 patients with type 2 diabetes (T2DM). Patients randomly divided into two groups to receive vitamin D (VD) supplementation (100 µg or 4000 IU/day) or placebo for three months, randomization was based on the permutated-block method. Anthropometric indices including body weight (BW), body mass index (BMI) and waist circumference (WC) and physical activity, dietary intake were assessed by validated methods at the beginning and end of the trial. RESULTS: VD supplementation had not any significant differences in anthropometric indices, dietary intake and physical activity between the two groups. CONCLUSION: Finally, it can be concluded, receiving 100 micrograms/day of VD for three months had no favourable effects on patients with T2DM.

High-Intensity Interval Training Reversed High-Fat Diet-Induced M1-Macrophage Polarization in Rat Adipose Tissue via Inhibition of NOTCH Signaling.

INTRODUCTION: There is accumulating evidence on the beneficial effect of exercise intervention in the management of metabolic disorders; however, the molecular mechanism is still unclear. Here, the current study aimed to compare the effect of high-intensity interval training (HIIT) and continuous endurance training (CET) on serum and adipose-tissue markers of M1/M2 macrophage polarization. METHODS: A total of 45 healthy male Wistar rats were divided into groups of normal chow (n=10) and high-fat diet (HFD) (n=35). Then, rats receiving the HFD were randomly divided into four groups. Training programs were performed for 5 days/week over 10 weeks. The CET protocol included 30 minutes running at 50%-60% of VO2max. The HIIT protocol consisted of five repeated intervals of 2-minute sprints on the treadmill at 80%-90% VO2max workload with 1 minute's 30%-35% VO2max interval for each rat. Then, biochemical parameters were assessed. Macrophage-polarization markers were assessed at mRNA and protein levels by real-time PCR and Western blotting, respectively. RESULTS: Both exercise-training programs, especially HIIT, reversed increased serum biochemical parameters (glucose, triglycerides, cholesterol, Homeostatic Model Assessment of Insulin Resistance, and hsCRP), M1-polarization markers (circulating IL6, TNFα, and adipose-tissue mRNA expression of IL6, TNFα and iNOS), M2 markers (CD206, CD163, and IL10 expression), as well as pIκKB, pNFκB, and NICD expression in HFD-induced diabetes. CONCLUSION: Our findings suggest that despite devoting less time, the HIIT workout is a more effective intervention for diabetes management. Moreover, HIIT reverses HFD-induced macrophage polarization by targeting the NFκB and NOTCH signaling pathways.

Cytotoxic effects and apoptosis induction of cisplatin-loaded iron oxide nanoparticles modified with chitosan in human breast cancer cells.

Cisplatin is widely used as an anticancer drug in chemotherapy of human cancers. In the field of cancer therapy, nanoparticles modified with biocompatible copolymers are suitable vehicles to effectively deliver smaller doses of hydrophobic drugs such as cisplatin in the body. In this study, we investigated whether cisplatin-loaded iron oxide nanoparticles (IONPs) modified with chitosan can exert cytotoxic effects in the human breast cancer cell line MDA-MB-231. IONPs was synthesized using eucalyptus leaf extract as a reducing and stabilizing agent. MDA-MB-231 cells were treated with different concentrations of cisplatin, cisplatin-IONPs and cisplatin-IONPs-chitosan for 24 h. Apoptosis was confirmed by flow cytometry, whereas The mRNA and protein expression of pro- and anti-apoptotic molecules were measured using Real time RT-PCR and western blotting. Treatment with both cisplatin-IONPs and cisplatin-IONPs-chitosan showed a significantly higher cytotoxic effect in comparison to the free drug alone in MDA-MB-231 cells. The levels of apoptosis in cells treated with a combination of cisplatin-IONPs-chitosan were significantly higher compared with cisplatin-IONPs and cisplatin alone. The results of this study showed that the interaction between cisplatin and iron oxide nanoparticles modified with chitosan could enhance responsiveness to cisplatin in breast cancer cells.

Synergistic anticancer activity of valproate combined with nicotinamide enhances anti-proliferation response and apoptosis in MIAPaca2 cells.

Histone deacetylase is strongly associated with epigenetic regulation and carcinogenesis, and its inhibitors can induce cell cycle arrest and apoptosis of the cancer cells. In this study we aimed to examine the antiproliferative effects a combination of the valproate with nicotinamide in MIAPaca2 cell line. We revealed that valproate acted in a synergistic/additive with nicotinamide to inhibit the proliferation and induction of apoptosis in MIAPaca2 cancer cell line. MIAPaca2 was treated with various concentrations of valproate. The MTT assay and colony formation in soft agar indicated that valproate at 0.5 mM, when used alone weakly, suppressed proliferation of cells (37 ± 3.02%) whereas the combination treatment of valproate + nicotinamide significantly suppressed cell proliferation (58 ± 3.5%). The effect of nicotinamide at 25 mM on cell proliferation and cell colonization induced 50% apoptosis of MIAPaca2 cells. To identify the anti-proliferation and apoptotic effects of valproate and nicotinamide we performed flow cytometric and microscopic analyses. The results indicated significant apoptosis induction and nuclear morphological alterations greater than when valproate was used alone. Furthermore, western blot analyses was performed to study the role of acetyl-histone H3 levels, and quantitative RNA expression analyses were performed on expression of thrombospondin (TSP) and maspin genes in MIAPaca2. We found that the combination treatment of valproate + nicotinamide enhanced the expression of maspin and TSP genes and the biological response of the cell line was correlated with the increase of histone H3 acetylation after nicotinamide and valproate application. Together our findings indicate that valproate which act as inhibitor of cell proliferation and inducer of apoptosis in human cancer MIAPaca2 cells when used in combination with nicotinamide makes it a potentially good candidate for new anticancer drug development.

The enhanced apoptosis and antiproliferative response to combined treatment with valproate and nicotinamide in MCF-7 breast cancer cells.

Acetylation of histone is a major player in epigenetic modifications, resulting in open chromatin structures and, hence, permissive conditions for transcription-factor recruitment to the promoters, followed by initiation of transcription. Histone deacetylase inhibitors arrest cancer cell growth and cause apoptosis with low toxicity thereby constituting a promising treatment for cancer. In this study, we examined the antiproliferative effects of valproate with a combination of nicotinamide in the MCF-7 cell line. MCF-7 was treated with various concentrations of valproate. The MTT assay showed that the viability of MCF-7 cells was inhibited and the cell activity was decreased. Viability percent of valproate and nicotinamide combined treatment cells (28 ± 2) was 1.78 times increased compared with the valproate-alone (0.5 mM) treated cells (50 ± 2). Colony formation in soft agar indicated that valproate at 0.3 mM, when used alone, weakly suppressed proliferation of cells (82 ± 3) and the combination treatment of valproate + nicotinamide strongly suppressed cell proliferation (51 ± 3). The flow cytometric and microscopic analyses of HDACI combined with treated cells indicated strong apoptosis induction and nuclear morphological alterations greater than those of valproate alone. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis confirmed the efficiency of the HDAC inhibitor combination, revealing the effectively upregulated p16 and p21. Furthermore, to investigate the role of acetyl-histone H3 levels, western blot analyses have been performed and high levels of acetylated histone H3 were detected in valproate- and nicotinamide-treated cells. These results suggest that the combination treatment of valproate with nicotinamide exerts significant antitumor activity and could be a promising therapeutic candidate to treat human breast cancer.

Differential effect of activin on mouse embryonic stem cell differentiation in insulin-secreting cells under nestin-positive selection and spontaneous differentiation protocols.

Parallel to the importance of the development of cell therapies to treat diabetes is the production of sufficient numbers of pancreatic endocrine cells that function like primary islets. To increase the efficiency of endocrine pancreatic-like cell differentiation from mouse embryonic stem cells (ESCs), we applied activin-B to nestin-positive selection (protocol 1) and spontaneous differentiation (protocol 2) in different groups including: [A] activin-B, or [B] basic fibroblast growth factor (bFGF), and/or [C] activin-B+bFGF. The differentiated cells expressed most pancreatic-related genes. The number of insulin- and C peptide-positive cells, as well as dithizone-positive clusters in group A of protocol 1 was higher than in the other groups. Significant insulin concentrations in protocol 1 were produced when glucose was added to the medium, in comparison with protocol 2. Moreover, insulin release was increased significantly in group A of protocol 1 even with lower glucose. In conclusion, Addition of activin-B in a nestin-positive selection protocol increased the insulin-secreting cells in comparison with the same protocol with bFGF and/or spontaneous differentiation in presence of bFGF and/or activin-B alone. However, improvements of the current method are required to generate a sufficient source of true beta-cells for the treatment of diabetes mellitus.

Generation of insulin-secreting cells from human embryonic stem cells.

A growth factor-mediated selection method was used to obtained insulin-secreting cells from human embryonic stem cells (hESC; Royan H1). Our resultant cells were positive for dithizone, a zinc-chelating agent known to selectively stain pancreatic beta cells and immunoreactive for antibodies against insulin, glucagon, and C-peptide. Semi-quantitative reverse transcription-polymerase chain reaction detected expression of proinsulin, insulin and other pancreatic beta-cell-related genes, such as Nkx6.1, Is11, Glut2, Pax4, and prohormone convertase2 (PC2). Moreover, glucagon, somatostatin, K(ATP)-channel genes KIR6.2 and SUR1, islet amyloid polypeptide (IAPP), PC1/3, and glucokinase (GCK) were expressed in the differentiating hESC in a developmental stage-dependent manner. Also, the addition of glucose to the culture medium triggered insulin release from differentiated cells, but transmission electron microscopy of the differentiated cells did not show typical beta-cell granules, even though secretary granules were detected. The results showed that hESC have the ability to transcribe and process insulin, but further improvements of the current method are required to generate a sufficient source of true beta cells for the treatment of diabetes mellitus.