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- ItemAcesso aberto (Open Access)Caracterização molecular de Oligozoospermia grave e Azoospermia não obstrutiva(Universidade Federal de São Paulo (UNIFESP), 2019-10-01) Vilela, Caio Graco Bruzaca Almeida [UNIFESP]; Fraietta, Renato [UNIFESP]; http://lattes.cnpq.br/5650852956005080; http://lattes.cnpq.br/1545035937368744; Universidade Federal de São Paulo (UNIFESP)Many cases of male infertility present an undetermined etiology, even after exhaus-tive clinical and laboratory investigations. An association between male infertility and the synaptonemic complex genes responsible for meiosis synapse is observed. In this context, this study aims to describe variants in genes related to the synap-tonemic complex. Next generation sequencing was performed comprising the SYCP1, SYCP2, SYCP2L and SYCP3 genes. A sample of 14 chromosomal males was assessed, categorized into four groups: nonobstructive azoospermic, cryptozoo-spermic, severely oligozoospermic and normozoothermic (control). No significant dif-ference was observed between the groups using dependent variables related to physical and hormonal examinations, except for mean testicular volume. Three vari-ants of uncertain significance were observed in the SYCP2 gene of the cryptozoo-spermic group. No pathogenic or potentially pathogenic variants were observed in the other assessed genes, both in controls and in patients presenting idiopathic male infertility. Thus, three new variants associated to the SYCP2 gene were added to male infertility cases and benign variants in normozoospermic fertile controls were confirmed.
- ItemAcesso aberto (Open Access)Efeitos do exercício resistido prévio ao infarto do miocárdio sobre a função cardíaca de ratas(Universidade Federal de São Paulo (UNIFESP), 2019-12-18) Silva, Flavio Andre [UNIFESP]; Tucci, Paulo Jose Ferreira [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)There is evidence that aerobic exercise prior to myocardial infarction (MI) is capable of generating cardioprotection. However, the cardioprotective properties of resistance exercise (RE) in infarcted rats have been little investigated. Objective: To evaluate the effects of RE prior to myocardial infarction (MI) on cardiac morphology, cardiac function, and proteins that participate in calcium kinetics, associating them with muscle function and cardiorespiratory fitness in female rats. Methods: Wistar rats were distributed into the following groups: SSh: sedentary prior to false surgery (Sham) (n=9); SIM: sedentary prior to MI (n=19); TIM: previously trained to IM (n=13). The exercise program consisted of four sets of 8 to 12 movements carrying weights corresponding to 80% of the maximum load (CM g) five days a week for eight weeks on a rodent-adapted ladder. At the end of the eighth week, the animals underwent MI or sham surgery and were analysed four weeks later. p< 0,05 was considered to be statistically significant. Results: The TIM group rats presented smaller MI (TIM: 43±6,3% e SIM: 55±8,2) and the scar of MI (TIM: 1.2±0,21cm e SIM: 1.6±0,28). Mortality showed no statistical difference (SIM: 37% e TIM: 39). Previous ER mitigated atrial mass increase (mg/mm) (SSh: 1,1±0,1 SIM: 3,8±0,7; TIM: 2,7±0,6), right ventricle (mg/mm) (SSh: 4,5±0,5; SIM: 8,5±2,5; TIM: 6,2±1,0) and heart (mg/mm) (SSh: 22±1; SIM: 29±4; TIM: 26±2) compared to the rats of the SIM group. Left ventricular weight (mg/mm) was similar between groups (SSh: 16,2±0,7, SIM: 16,2±1,5; TIM: 16,9±0,8). The previous RE attenuated the enlargement of the left atrium area (cm²) (SSh: 0,37±0,03; SIM: 0,66±0,09; TIM: 0,52±0,13), wave E (m/s) (SSh: 0,86±0,09; SIM: 1,1±0,13; TIM: 0,97±0,16) and E/A ratio (SSh: 2,9±0,8; SIM: 5,9±1,7; TIM: 4,3±1,8) compared to the rats of the SIM group. The pulmonary water content (SSh: 79,8±0,27%; SIM: 80,71±1,7; TIM: 82,01±2,1) and hepatic (SSh: 70,56±0,1%; SIM: 70,34±0,9; TIM: 71,37±0,7) was superior in TIM. RE slowed down skeletal muscle mass decline (SSh: 0,12±0,01g; SIM: 0,13±0,01; TIM:0,15±0,04). The VO2pico (ml/kg/min) (SSh: 58±6; SIM: 51±10; TIM: 52±6) and maximum speed (Vmáx cm/s) (SSh: 85±12; SIM: 51±10; TIM: 52±6) were inferior in the infarcted. The protein expression (%Sham) of calcium sodium exchanger (NCX) (SSh: 100±32; SIM: 102±29; TIM: 123±39) and total phospholambam (tPLB) (SSh: 100±20; SIM: 86±8; TIM: 86±17) were not different between groups. The protein expression (%Sham) of SERCA2a (SSh: 100±13; SIM: 65±17; TIM:62±14,43) and phosphophorylated phospholambam (pPLB) (SSh: 100±32; SIM: 66±34; TIM: 58±23) were lower in infarcted animals. Conclusion: The RE performed prior to myocardial infarction minimized the size of the MI, alleviated left atrial dissection, attenuated myocardial hypertrophy, and increased muscle mass associated with better strength performance of the skeletal muscles.
- ItemAcesso aberto (Open Access)Vitamina D: a influência na matriz extracelular do tecido conjuntivo de sustentação dos órgãos pélvicos(Universidade Federal de São Paulo (UNIFESP), 2019-11-01) Kemp, Marta Maria [UNIFESP]; Girao, Manoel Joao Batista Castello [UNIFESP]; Nader, Helena Bonciani [UNIFESP]; http://lattes.cnpq.br/7175631659428994; http://lattes.cnpq.br/0973903299568770; Universidade Federal de São Paulo (UNIFESP)Hypothesis/aims of study: Vitamin D deficiency is largely spoken of as being associated with several different diseases. Most of the associations are inferred by the levels of 25(OH)D in the plasma, which has no direct correlation to the activity of its active metabolites, i.e. 1,25(OH)D, since it follows the patterns of a steroid hormone, with negative and positive feedbacks and other molecular interactions, instead of behaving as a vitamin. Pelvic organ prolapse has also been linked to lower serum levels of vitamin D, although the exact mechanism for this increase in risk is not totally understood. One of the potential mechanisms for this increased risk is the influence vitamin D has on the architecture of the extracellular matrix (ECM) main component of the structures of the pelvic support apparatus and responsible for its mechanical properties. The aim of this study was to examine the arrangement of collagen fibers, the most prevalent fibrous protein present in the ECM at the endopelvic fascia, and other structural components of the ECM and cytoskeleton of the endopelvic fascia, to capture any different morphological pattern between subjects with a normal diet and those under a vitamin D supplementation. Collagen quantification was also performed. Study design, materials and methods: This hypothesis was investigated using female healthy adult Wistar rats (8 weeks old, approximately 200-220g). The treatment group received cholecalciferol diluted in arachid oil by gavage (in the dose 37.5mcg/Kg/day, equivalent to 1,500Ui/Kg/day), and the control group received only the oil, for 18 consecutive days. Twenty-four hours after the last gavage procedure, euthanasia was performed and a surgical procedure was undertaken extracting, as a bloc, the lower third of the uterus with the uterosacral ligament, the upper half of the vagina with the connective tissue surrounding and the bladder with proximal urethra. Samples were immersed in Tissue-Tek and cut in 30 and 50 micrometer thick slices and placed on the sheets for microscopic evaluation with SHG (second harmonic generation) technique, which requires no tissue preparation. For the purpose of interpretation, the rats were also separated into groups according to the estrous phase: high estrogenic influence (proestrus and estrus) and low estrogenic influence (metaestrus and diestrus) identified by cytology and histology under conventional light microscopy. Collagen by Picrosirius and antibodies for other extracellular structural components were also performed. Tissues were analyzed using Confocal Microscope (TCS SP8 CARS from Leica) and F and Epi-SHG detectors were used to detect SHG signals from collagen fibers. Images are represented as maximum intensity projections, corresponding to the Z-series of confocal stacks and were collected by tile scan and processed with Leica LAS AF software and, posteriorly, manipulated with ImageJ software. Results: We found that collagen fibers have a consistently undulated pattern and more random disposition in the xvi treated group whereas in the control group it assumes a more taut and oriented appearance, when under high estrogenic influence. The collagen signal from SHG was statistically more intense in the treated group compared to the control group also under a high estrogenic environment. The extracellular components alpha-actin, heparan sulfate and glicoconjugates containing sialic acid and N-acetylgkucosamine residues also showed clear different images when comparing control and treated groups. Conclusion: These findings suggest that there is a difference in the morphological presentation of the extracellular matrix of subjects under high dose intake of vitamin D compared to subjects under normal intake of the vitamin, pointing to a probable change in the mechanical properties of the tissue and that there might be an interaction of the metabolic pathways of the two steroid hormones cholecalciferol and estrogen at least regarding the amount of collagen deposited in the tissue. Not only the collagen, other components of the ECM presented differently in the treated and control groups. Oral high dose intake of vitamin D changes the framework and composition of the extracellular matrix at the level of the endopelvic fascia, which can impact the properties and resilience of the tissue. In summary, new research are needed in the biomechanical field to approach a clinical significance but we have shown a tangible relation between vitamin D exposure and collagen fiber architecture.