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INTRODUCTION: Imaginary exercises seem to be useful therapeutic approaches to modulate neuromuscular functions due to two main reasons: first, this training would not greatly increase body temperature, and secondly, it can positively affect brain-muscle pathways-which are both primary factors should be considered in rehabilitation programs for patients with multiple sclerosis (MS). METHOD: 32 pre-elderly adult females with relapsing-remitting MS (n = 16 - age M (SD): 56.75 (5.07)) and without MS (n = 16 - age M (SD): 56.56 (4.35)) voluntarily recruited. First, they were assigned into two groups: MS patients and healthy controls, to investigate baseline between-group comparison. Then, MS patients were randomly divided into two groups of eight each, designated as experimental and control groups. Recording the nerve conduction velocity (NCV) of tibial nerve and integrated electromyographic muscle activation (IEMG) of gastrocnemius muscle was conducted twice, before and after a six-week mind-body exercise therapy to evaluate its effectiveness on improving neuromuscular function. RESULTS: The results showed significant difference in both tibial NCV (P < 0.001) and IEMG (P = 0.001) variables between non-MS group and MS group. Furthermore, there was a significant main effect of intervention (P = 0.05) and time (P < 0.001) on IEMG in the MS group, while there was no significant effect of intervention (P = 0.18) and time (P = 0.23) on NCV (p = 0.89). CONCLUSION: Neuromuscular dysfunction were apparent in MS patients, and a mind-body therapy of imagery isometric training was found to be useful on improving the neurological deficit in women with MS. TRIAL REGISTRATION NUMBER: UMIN000046935.
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Eletromiografia , Terapias Mente-Corpo , Esclerose Múltipla Recidivante-Remitente , Músculo Esquelético , Humanos , Feminino , Pessoa de Meia-Idade , Músculo Esquelético/fisiologia , Músculo Esquelético/fisiopatologia , Esclerose Múltipla Recidivante-Remitente/reabilitação , Esclerose Múltipla Recidivante-Remitente/fisiopatologia , Terapias Mente-Corpo/métodos , Condução Nervosa/fisiologia , Nervo Tibial/fisiologia , Pandemias , AdultoRESUMO
Background: It is unclear whether resistance training in combination with different timing of protein intake might have differential effects on muscle hypertrophy, strength, and performance. Therefore, we compared the effects of 8 weeks of resistance training combined with two different high-protein diet strategies (immediately pre-and after, or 3 h pre and after exercise) in resistance-trained males. Methods: Forty resistance-trained males (24 ± 4 years) performed 8 weeks of resistance training combined with 2 g kg-1 d-1 protein. Body composition, muscular performance, and biochemical markers were assessed pre and post-intervention. Results: Nine participants (four from 3 h group and five from the immediate group) withdrew from the study. Therefore, 31 participants completed the study. All measures of skeletal muscle mass, Australian pull-up, and muscle strength, significantly increased post-intervention in both groups (p < 0.05). The biochemical marker urea also significantly increased from pre to post in both groups (p < 0.05). There were no significant between-group differences (p > 0.05). Conclusion: High-protein diet enhances muscular performance and skeletal muscle mass in resistance-trained males, irrespective of intake time. Consequently, the total daily protein intake appears to be the primary factor in facilitating muscle growth induced by exercise.
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Empagliflozin as an antioxidant decreases blood glucose and insulin resistance in type 2 diabetes mellitus. Base on the empagliflozin antioxidant properties we decided to investigate the its effects on the testis histological changes through stereological techniques and biochemical evaluations in T2 diabetes mellitus rats. Rats were divided into: control, diabetes mellitus (DM, streptozotocin + nicotinamide) and diabetes mellitus + empagliflozin (DM + EMPA, 10 mg/kg/day) groups. 56 days after inducing diabetes mellitus testis histological changes and serum biochemical factors along with the level of Bax, Bcl2 and Nrf2 genes expression in the testicular tissue were assessed. A significant decrease in the mean total volume of testis and its components, the level of Bcl2 and Nrf2 gene expression (p < 0.001) along with a significant increase in the level of IL-6, TNF-α, MDA, Bax gene expression were observed in the DM group compared to the control group (p < 0.001). In the DM + EMPA group, the mean total volume of testis and its components, the level of Bcl2 gene expression (p< 0.01) and Nrf2 (p < 0.001) significantly increased whereas the mean level of IL-6 (p < 0.01), TNF-α (p < 0.001), MDA (p < 0.001), Bax (p < 0.001) gene expression significantly decreased compared to the DM group. Our results showed that empagliflozin, by improving the antioxidant defense system, can reduce testicular inflammation and apoptosis and partly prevent the adverse effects of diabetes mellitus on testicular tissue.
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Compostos Benzidrílicos , Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Glucosídeos , Masculino , Ratos , Animais , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Tipo 2/tratamento farmacológico , Antioxidantes/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Interleucina-6/metabolismo , Proteína X Associada a bcl-2/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Estresse OxidativoRESUMO
CONTEXT: Ovarian tissue transplantation is performed to preserve fertility in patients undergoing chemotherapy and radiotherapy. However, the ischemia-reperfusion injury which occurs after the ovarian tissue transplantation causes follicular depletion and apoptosis. l -Carnitine has antioxidant and anti-inflammation properties. AIMS: Therefore, we aimed to investigate the beneficial effect of l -carnitine on mouse ovaries following heterotopic autotransplantation. METHODS: Mice were randomly divided into three groups (six mice per group): control, autografted and autografted+l -carnitine (200mg/kg daily intraperitoneal injections). Seven days after ovary autografting, the serum levels of malondialdehyde (MDA), total antioxidant capacity, tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-10 were measured. Ovary histology, serum concentrations of progesterone and estradiol were also measured 28days after autotransplantation. Data were analysed using one-way analysis of variance (ANOVA) and Tukey test, and the means were considered significantly different at P Key results: In the autografted+l -carnitine group, the total volume of the ovary, the volume of the cortex, the number of follicles, the serum concentrations of IL-10, estradiol and progesterone significantly increased compared to the autografted group. In the autografted+l -carnitine group, serum concentrations of IL-6, TNF-α and MDA were significantly decreased compared to the autografted group. CONCLUSIONS: Our results indicated that l -carnitine can ameliorate the consequences of ischemia-reperfusion on the mice ovarian tissue following autotransplantation. IMPLICATIONS: l -carnitine improves the structure and function of transplanted ovaries.
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Carnitina , Ovário , Animais , Feminino , Humanos , Camundongos , Antioxidantes , Carnitina/farmacologia , Carnitina/uso terapêutico , Estradiol , Interleucina-10 , Interleucina-6 , Ovário/patologia , Progesterona , Fator de Necrose Tumoral alfaRESUMO
A growing body of evidence suggests voluntary physical activity is associated with decreased stress-related disorders such as anxiety- and depression-like behaviours in both humans and rodents. The postpartum period is also a vulnerable transition time for the development of these neurobehavioural disorders in women. This study aimed to determine whether voluntary physical activity during pregnancy and postpartum period can increase maternal care and decrease anxiety- and depression-related behaviours in postpartum dams. To this end, pregnant mice were exposed to running wheel during their gestational and postpartum periods, and then nest building, active nursing, and licking/grooming behaviours were recorded as maternal care. To assess depression and anxiety-related symptoms, several behavioural tests such as the novelty-suppressed feeding test, tail suspension test, sucrose preference test, social interaction test, forced swim test, open field, elevated plus maze, light-dark box, and elevated zero maze were used. To identify the most important mechanisms behind these behavioural alterations, we measured oxytocin, adrenocorticotropic hormone (ACTH) and corticosterone in the serum and serotonin in the brain of postpartum dams. Our findings showed that running wheel significantly increased maternal care, and decreased depression-like behaviour during the postpartum period through increasing serum oxytocin and brain serotonin levels, whereas it decreased anxiety-like behaviour via attenuating the hypothalamic-pituitaryadrenal (HPA) axis activity by measuring ACTH and corticosterone levels in postpartum dams. Overall, this study suggests that voluntary physical activity during pregnancy and the postpartum period might improve maternal care and decrease anxiety and depression-related behaviours in postpartum dams.
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Corticosterona , Depressão , Hormônio Adrenocorticotrópico , Animais , Ansiedade , Comportamento Animal , Feminino , Humanos , Comportamento Materno , Camundongos , Ocitocina , Período Pós-Parto , Gravidez , Serotonina , Estresse PsicológicoRESUMO
Metformin has long been used in the treatment of polycystic ovarian syndrome (PCOS). Recently, sitagliptin has been reported to improve ovarian cycles and ovulation in PCOS. We suggest that a combination of sitagliptin and metformin can be more effective than either treatment alone in improving different aspects of PCOS.
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Metformina/uso terapêutico , Fosfato de Sitagliptina/uso terapêutico , Feminino , Humanos , Ciclo Menstrual/fisiologia , Ovulação/fisiologia , Síndrome do Ovário PolicísticoRESUMO
Polycystic ovary syndrome (PCOS) is related to low levels of serum l-carnitine, which has antioxidant, anti-inflammatory and antiapoptotic properties. The aim of this study was to investigate the effect of l-carnitine on folliculogenesis in mice following induction of PCOS. PCOS was induced by daily injections of testosterone enanthate (1mg per 100g, s.c., for 35 days). NMRI mice (21 days old) were divided into four groups (n=6 per group): Control, Control+l-carnitine, PCOS and PCOS+l-carnitine. Mice were treated with 500mgkg-1, i.p., l-carnitine every second day for 28 days. Ovaries were studied stereologically and serum concentrations of FSH, LH, testosterone, interleukin (IL)-6 and tumour necrosis factor (TNF)-α were determined using ELISA kits. Serum concentrations of malondialdehyde (MDA) and the ferric ion reducing antioxidant power (FRAP) were also analysed. Apoptosis of follicles was evaluated by terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL). CD31 was assessed immunohistochemically. Data were analysed using one-way analysis of variance (ANOVA) and Tukey's test, differences considered significant at P<0.05.The total volume of the ovary, cortex volume, oocyte volume, zona pellucida thickness and the number of antral follicles increased significantly, whereas the number of primary and preantral follicles decreased significantly, in the PCOS+l-carnitine versus PCOS group. In the PCOS+l-carnitine group, serum concentrations of FSH and FRAP increased significantly, whereas there were significant decreases in serum concentrations of testosterone, LH, MDA, IL-6 and TNF-α, as well as in the percentage of TUNEL-positive apoptotic cells, compared with the PCOS group. l-Carnitine improves folliculogenesis and is therefore suggested as a therapeutic supplement in the treatment of PCOS.
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Apoptose/efeitos dos fármacos , Carnitina/farmacologia , Inflamação/tratamento farmacológico , Folículo Ovariano/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Síndrome do Ovário Policístico/tratamento farmacológico , Animais , Apoptose/fisiologia , Carnitina/uso terapêutico , Modelos Animais de Doenças , Feminino , Hormônio Foliculoestimulante/sangue , Inflamação/metabolismo , Interleucina-6/sangue , Hormônio Luteinizante/sangue , Malondialdeído/sangue , Camundongos , Folículo Ovariano/metabolismo , Ovário/efeitos dos fármacos , Ovário/metabolismo , Estresse Oxidativo/fisiologia , Síndrome do Ovário Policístico/metabolismo , Testosterona/sangue , Fator de Necrose Tumoral alfa/sangueRESUMO
BACKGROUND: Many studies have reported that inflammation and oxidative stress are involved in the pathogenesis of polycystic ovary syndrome (PCOS). Bone marrow mesenchymal stromal cells (BM-MSCs) have anti-oxidant and anti-inflammation properties. In this study, we investigate the beneficial effect of stem cell therapy on folliculogenesis in mice with induced PCOS METHODS: Mouse model of PCOS was performed through daily injection of testosterone enanthate (1 mg/100 g/body weight subcutaneous (s.c).) for a period of 5 weeks. Naval Medical Research Institute (NMRI) mice (21 days old) were divided into three groups: control, PCOS and PCOSâ¯+â¯BM-MSCs. BM-MSCs were labeled with Hoechst 33342 (0.5 µg/mL) and then injected into the mice (106/animal, via the tail vein) at 1 and 14 days after PCOS confirmation. Mice were humanely killed at 2 weeks after last transplantation. Ovarian stereological studies were done. Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), testosterone, interleukin (IL)-6 and tumor necrosis factor (TNF)-α serum levels were measured. The levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) in serum were analyzed. Apoptotic index for ovarian follicles was assessed using Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL). CD31 expression in ovarian vessels was assessed with the immunohistochemistry. RESULTS: There was a significant increase in the total volume of ovary, cortex, number of antral follicles, volume of oocyte and zona pellucida thickness, and there was a significant decrease in the primary and preantral follicles number in the PCOSâ¯+â¯BM-MSCs group compared with the PCOS group. There was a significant increase in the serum level of FSH and TAC and a significant decrease in the serum level of testosterone, LH, MDA and percentage of TUNEL-positive apoptotic cells in the PCOSâ¯+â¯BM-MSCs group in comparison with the PCOS group. DISCUSSION: BM-MSC transplantation improves folliculogenesis in mice with induced PCOS. BM-MSC therapy can be an operative treatment for PCOS via anti-inflammatory, anti-oxidant and anti-apoptotic properties.
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Transplante de Células-Tronco Mesenquimais/métodos , Folículo Ovariano/fisiologia , Ovário/fisiologia , Síndrome do Ovário Policístico/terapia , Animais , Antioxidantes/metabolismo , Medula Óssea , Modelos Animais de Doenças , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Luteinizante/sangue , Malondialdeído/metabolismo , Células-Tronco Mesenquimais/citologia , Camundongos , Oócitos/citologia , Oócitos/fisiologia , Ovário/citologia , Síndrome do Ovário Policístico/induzido quimicamente , Testosterona/análogos & derivados , Testosterona/toxicidade , Fator de Necrose Tumoral alfa/sangueRESUMO
Human mesenchymal stem cells were exposed to diabetic sera for 7 days. Cell viability and apoptosis rate were detected by MTT and flow cytometry assays. The expression of key genes such as CD63, Alix, Rab27a, Rab27b, and Rab8b was monitored by real-time PCR. We also measured acetylcholinesterase activity and size and zeta potential of exosomes in the supernatant form diabetic cells and control. The cellular distribution of CD63 was shown by immunofluorescence imaging and western blotting. Any changes in the ultrastructure of cells were visualized by electron microscopy. Data showed a slight decrease in survival rate and an increased apoptosis in diabetic cells as compared to control (p < 0.05). By exposing cells to diabetic sera, a significant increase in the level of all genes CD63, Alix, Rab27a, Rab27b, and Rab8b was observed (p < 0.05). Flow cytometry analysis and immunofluorescence imaging confirmed increasing CD63 protein content upon treatment with diabetic sera (p < 0.05). We found an enhanced acetylcholinesterase activity in a diabetic condition which coincided with the increasing size of exosomes and decrease in zeta potential (p < 0.05). The fatty acid profile was not significantly affected by diabetic sera. Ultrastructural examination detected more accumulated cytoplasmic lipid vacuoles in diabetic cells.
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Diabetes Mellitus/sangue , Diabetes Mellitus/metabolismo , Exossomos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Transdução de Sinais , Acetilcolinesterase/metabolismo , Apoptose , Sobrevivência Celular , Exossomos/ultraestrutura , Ácidos Graxos/metabolismo , Humanos , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Eletricidade Estática , Tetraspanina 30/metabolismo , Regulação para Cima/genéticaRESUMO
This experiment investigated the impact of serum from patients with type 2 diabetes mellitus on the angiogenic behavior of human mesenchymal stem cells in vitro. Changes in the level of Ang-1, Ang-2, cell migration, and trans-differentiation into pericytes and endothelial lineage were monitored after 7 days. The interaction of mesenchymal stem cells with endothelial cells were evaluated using surface plasmon resonance technique. Paracrine restorative effect of diabetic stem cells was tested on pancreatic ß cells. Compared to data from FBS and normal serum, diabetic serum reduced the stem cell survival and chemotaxis toward VEGF and SDF-1α (P < 0.05). Diabetic condition were found to decline cell migration rate and the activity of MMP-2 and -9 (P < 0.05). The down-regulation of VEGFR-2 and CXCR-4 was observed with an increase in the level of miR-1-3p and miR-15b-5p at the same time. The paracrine angiogenic potential of diabetic stem cells was disturbed via the changes in the dynamic of Ang-1, Ang-2, and VEGF. Surface plasmon resonance analysis showed that diabetes could induce an aberrant increase in the interaction of stem cells with endothelial cells. After treatment with diabetic serum, the expression of VE-cadherin and NG2 and ability for uptake of Dil-Ac-LDL were reduced (P < 0.01). Conditioned media prepared from diabetic stem cells were unable to decrease fatty acid accumulation in ß-cells (P < 0.05). The level of insulin secreted by ß-cells was not affected after exposure to supernatant from diabetic or non-diabetic mesenchymal stem cells. Data suggest diabetes could decrease angiogenic and restorative effect of stem cells in vitro. J. Cell. Biochem. 119: 524-535, 2018. © 2017 Wiley Periodicals, Inc.
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Diabetes Mellitus Tipo 2/sangue , Células-Tronco Mesenquimais/metabolismo , Neovascularização Fisiológica , Soro , Idoso , Angiopoietina-1/biossíntese , Angiopoietina-2/biossíntese , Humanos , Masculino , Metaloproteinase 2 da Matriz/biossíntese , Pessoa de Meia-Idade , Receptores CXCR4/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/biossínteseRESUMO
Polycystic ovary syndrome (PCOS) is associated with low-quality oocytes. The aim of the present study was to investigate the effects of metformin (MET), N-acetylcysteine (NAC) and their combination on follicular fluid parameters, oocytes and embryo quality in PCOS patients. A prospective randomised placebo-controlled pilot study on 60 Iranian women with PCOS (aged 25-35 years) undergoing intracytoplasmic sperm injection (ICSI) was designed. Women were divided into four groups (n=15 in each): (1) an MET, administered 1500mg day(-1) MET; (2) an NAC group, administered 1800mg day(-1) NAC; (3) an NAC + MET group; and (4) a placebo group. Drugs were administered from the 3rd day of previous cycle until the day of oocyte aspiration (6 weeks treatment in total). Data were analysed by one-way ANOVA, with significance set at P<0.05. The number of immature and abnormal oocytes decreased significantly in the NAC compared with placebo group, with a concomitant increase in the number of good-quality embryos in the NAC group (P<0.05). Malondialdehyde levels decreased significantly in the NAC and NAC + MET groups compared with the placebo-treated group (P<0.02). In addition, there were significant decreases in leptin levels in the NAC, MET and NAC + MET groups compared with the placebo group (P<0.001). Insulin and LH levels were significantly lower in the MET and NAC groups compared with the placebo-treated group (P<0.02). We concluded that NAC improves oocyte and embryo quality and could be administered as an alternative to MET.
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Acetilcisteína/uso terapêutico , Antioxidantes/uso terapêutico , Ectogênese/efeitos dos fármacos , Infertilidade Feminina/prevenção & controle , Oogênese/efeitos dos fármacos , Síndrome do Ovário Policístico/tratamento farmacológico , Injeções de Esperma Intracitoplásmicas , Centros Médicos Acadêmicos , Acetilcisteína/efeitos adversos , Adulto , Anti-Inflamatórios não Esteroides/efeitos adversos , Anti-Inflamatórios não Esteroides/uso terapêutico , Antioxidantes/efeitos adversos , Quimioterapia Combinada , Feminino , Líquido Folicular/efeitos dos fármacos , Humanos , Hipoglicemiantes/efeitos adversos , Hipoglicemiantes/uso terapêutico , Infertilidade Feminina/etiologia , Irã (Geográfico) , Metformina/efeitos adversos , Metformina/uso terapêutico , Recuperação de Oócitos , Oócitos/efeitos dos fármacos , Oócitos/patologia , Indução da Ovulação , Pacientes Desistentes do Tratamento , Projetos Piloto , Síndrome do Ovário Policístico/fisiopatologiaRESUMO
BACKGROUND: Studies have demonstrated the efficacy of metformin (MTF ) in reducing insulin resistance and N-acetyl cysteine (NAC) in inhibiting oxidative stress which are involved in the pathogenesis of polycystic ovarian syndrome (PCOS). We aimed to compare the effects of MTF and NAC combination on serum metabolite and hormonal levels during the course of ovulation induction in PCOS individual candidates of intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: In this prospective randomized clinical trial, placebo con- trolled pilot study, 80 patients of polycystic ovarian syndrome at the age of 25-35 years were divided into 4 groups (n=20): i. NAC=treated with N-acetyl cysteine (600 mg three times daily), ii. MTF=treated with metformin (500 mg three times daily), iii. MTF+NAC=treated with N-acetyl cysteine plus metformin (the offered doses) and iv. placebo (PLA). A total number of 20 patients (6 from MTF group, 4 from NAC group, 6 from MTF+NAC group and 4 from PLA group) were dropped of the study. The drugs were administrated from day 3 of menses of previous cycle until ovum pick-up. RESULTS: Serum levels of luteinizing hormone (LH), total testosterone, cholester- ol and triglyceride, insulin and leptin significantly reduced in the MTF and NAC groups compared to the placebo (p<0.01). But levels of LH, total testosterone, cholesterol and triglyceride had no significant reduction in the MTF+NAC groups compared to the placebo. The serum levels of malonyldialdehyde (MDA), insulin and leptin reduced significantly after treatment in the MTF+NAC group compared to the placebo (p<0.05). CONCLUSION: CONSIDERING THE ADVERSE EFFECT OF COMBINATION THERAPY, WE PROPOSED THE CONADMINISTRATION MIGHT HAVE NO BENEFICIAL EFFECT FOR PCOS PATIENT DURING COURSE OF OVULATION INDUCTION OF ICSI (REGISTRATION NUMBER: IRCT201204159476N1).
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OBJECTIVES: Consumption of high-fat foods is one of the major causes of obesity. Physical exercise is a strategy used to counteract obesity. The aim of this study was to investigate the effect of eight weeks endurance training and high-fat diet (HFD) on appetite-regulating hormones in rat plasma. MATERIALS AND METHODS: TWENTY EIGHT MALE WISTAR RATS WERE RANDOMLY DIVIDED INTO FOUR GROUPS: Control group with standard diet (CSD), endurance training with a standard diet (ESD), control group with high-fat diet (CHFD) and endurance training with high-fat diet (EHFD). Twenty-four hr after the last training session, the blood samples were obtained and analyzed for hormones levels. RESULTS: The significant increased weight gain and food intake and decreased plasma nesfatin-1 and PYY3-36 levels were observed in CHFD group, while exercise under the HFD antagonized these effects. There were no significant changes in ghrelin, insulin and leptin levels in different groups. CONCLUSION: These results suggest that exercise can prevent fattening effect of HFD. Probably, performing exercise makes a reduction of food intake and weight gain in rat via the increase in nesfatin-1 and PYY levels. However, further studies are necessary to understand the exact mechanisms involved in this field.
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OBJECTIVE: Sensory neurons in dorsal root ganglia (DRG) undergo apoptosis after peripheral nerve injury. The aim of this study was to investigate sensory neuron death and the mechanism involved in the death of these neurons in cultured DRG. MATERIALS AND METHODS: In this experimental study, L5 DRG from adult mouse were dissected and incubated in culture medium for 24, 48, 72 and 96 hours. Freshly dissected and cultured DRG were then fixed and sectioned using a cryostat. Morphological and biochemical features of apoptosis were investigated using fluorescent staining (Propidium iodide and Hoechst 33342) and the terminal Deoxynucleotide transferase dUTP nick end labeling (TUNEL) method respectively. To study the role of caspases, general caspase inhibitor (Z-VAD.fmk, 100 µM) and immunohistochemistry for activated caspase-3 were used. RESULTS: After 24, 48, 72 and 96 hours in culture, sensory neurons not only displayed morphological features of apoptosis but also they appeared TUNEL positive. The application of Z-VAD.fmk inhibited apoptosis in these neurons over the same time period. In addition, intense activated caspase-3 immunoreactivity was found both in the cytoplasm and the nuclei of these neurons after 24 and 48 hours. CONCLUSION: Results of the present study show caspase-dependent apoptosis in the sensory neurons of cultured DRG from adult mouse.
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OBJECTIVES: Para-Nonylphenol (p-NP) is used in many industries and our previous study showed that p-NP causes a reduction in rats bone marrow mesenchymal stem cells (MSCs) viability. The aim of this study was to investigate the effect of p-NP on osteogenic differentiation of MSCs. MATERIALS AND METHODS: MSCs were isolated and expanded to 3rd passage, then cultured in DMEM supplemented with osteogenic media as well as 0.5 or 2.5 µM of p-NP. After 5, 10, 15, and 21 days, the viability and the level of mineralization was determined using MTT assay and alizarin red, respectively. In addition, morphology and nuclear diameter of the cells were studied with the help of fluorescent dye. Furthermore, calcium content and alkalinphosphatase activity were also estimated using commercial kits. Data were statistically analyzed and the P<0.05 was taken as the level of significance. RESULTS: The viability and mineralization of the cells treated with 2.5 µM of p-NP reduced significantly after day 10 in comparison with the control group and administration of 0.5 µM. Moreover, chromatin condensation, reduction of nuclei diameter, and cytoplasm shrinkage was observed in the cell treated with 2.5 µM. The calcium concentration and alkalinphosphatase activity of the cells decreased significantly with 2.5 µM of p-NP when compared with 0.5 µM and control group. CONCLUSION: Adverse effect of p-NP was observed on osteogenic differentiation of MSCs at 2.5 µM due to disruption of mineralization. We strongly suggest more investigations on this chemical with respect to other stem cells, especially skin stem cells as p-NP is used in the formulation of cosmetics.
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There is much evidence that allergic symptoms represent a major health problem in polluted cities. The aim of this research is to elucidate some microscopic effects of air pollutants on pollen structure, proteins, and allergenicity. A scanning electron microscopy study of pollen grains indicated that in polluted areas, airborne particles accumulate on the surface of pollen grains and change the shape and tectum of pollen. Also, many vesicles are released from polluted pollen grains and the pollen material agglomerates on the surface of pollen grains. SDS-PAGE revealed that different proteins exist in mature and immature pollen grains. There were no significant differences between protein bands of polluted and nonpolluted pollen grains, but in polluted pollen, protein content decreases in response to air pollution, causing the release of pollen proteins. The results indicate that mature pollen have more allergenicity than immature pollen. According to the experiments polluted pollen grains are more effective than nonpolluted pollen grains in inducing allergic symptoms. Air pollutants can cause allergic symptoms, but when associated with allergen pollen grains, their allergenicity power is increased.