Isolation and primary culture of human abdominal aorta smooth muscle cells from brain-dead donors: an experimental model for vascular diseases.

Primary cell cultures are essential tools for elucidating the physiopathological mechanisms of the cardiovascular system. Therefore, a primary culture growth protocol of cardiovascular smooth muscle cells (VSMCs) obtained from human abdominal aortas was standardized. Ten abdominal aorta samples were obtained from patients diagnosed with brain death who were organ and tissue donors with family consent. After surgical ablation to capture the aorta, the aortic tissue was removed, immersed in a Custodiol® solution, and kept between 2 and 8 °C. In the laboratory, in a sterile environment, the tissue was fragmented and incubated in culture plates containing an enriched culture medium (DMEM/G/10% fetal bovine serum, L-glutamine, antibiotics and antifungals) and kept in an oven at 37 °C and 5% CO2. The aorta was removed after 24 h of incubation, and the culture medium was changed every six days for twenty days. Cell growth was confirmed through morphological analysis using an inverted optical microscope (Nikon®) and immunofluorescence for smooth muscle alpha-actin and nuclei. The development of the VSMCs was observed, and from the twelfth day, differentiation, long cytoplasmic projections, and adjacent cell connections occurred. On the twentieth day, the morphology of the VSMCs was confirmed by actin fiber immunofluorescence, which is a typical characteristic of VSMCs. The standardization allowed VSMC growth and the replicability of the in vitro test, providing a protocol that mimics natural physiological environments for a better understanding of the cardiovascular system. Its use is intended for investigation, tissue bioengineering, and pharmacological treatments.

Short-Term High-Fat Diet Increases Leptin Activation of CART Neurons and Advances Puberty in Female Mice.

Leptin is a permissive factor for puberty initiation, participating as a metabolic cue in the activation of the kisspeptin (Kiss1)-gonadotropin-releasing hormone neuronal circuitry; however, it has no direct effect on Kiss1 neurons. Leptin acts on hypothalamic cocaine- and amphetamine-regulated transcript (CART) neurons, participating in the regulation of energy homeostasis. We investigated the influence of a short-term high-fat diet (HFD) on the effect of leptin on puberty timing. Kiss1-hrGFP female mice received a HFD or regular diet (RD) after weaning at postnatal day (PN)21 and were studied at PN28 and PN32. The HFD increased body weight and plasma leptin concentrations and decreased the age at vaginal opening (HFD, 32 ± 0.53 days; RD, 38 ± 0.67 days). Similar colocalization of neurokinin B and dynorphin in Kiss1-hrGFP neurons of the arcuate nucleus (ARC) was observed between the HFD and RD groups. The HFD increased CART expression in the ARC and Kiss1 messenger RNA expression in the anteroventral periventricular (AVPV)/anterior periventricular (Pe). The HFD also increased the number of ARC CART neurons expressing leptin-induced phosphorylated STAT3 (signal transducer and activator of transcription 3) at PN32. Close apposition of CART fibers to Kiss1-hrGFP neurons was observed in the ARC of both RD- and HFD-fed mice. In conclusion, these data reinforce the notion that a HFD increases kisspeptin expression in the AVPV/Pe and advances puberty initiation. Furthermore, we have demonstrated that the HFD-induced earlier puberty is associated with an increase in CART expression in the ARC. Therefore, these data indicate that CART neurons in the ARC can mediate the effect of leptin on Kiss1 neurons in early puberty induced by a HFD.

Importância da atuação do Laboratório de Saúde Pública nas perícias judiciais relacionadas a matérias estranhas em alimentos / Importance of the Public Health Laboratory on the expert advice requested by the courts regarding foreign matter in food

O Instituto Adolfo Lutz (IAL) atua de maneira relevante em processos judiciais realizando perícias laboratoriais conhecidas como produção antecipada de provas. Este trabalho faz o relato da investigação da suspeita de presença de cacos de vidro em vinho tinto, referente a um processo judicial de município da região de Ribeirão Preto/SP. Em abril de 2013, uma amostra de vinho foi encaminhada ao IAL-Ribeirão Preto para confirmação de corpo estranho e sua identificação pelo Laboratório de Microscopia de Alimentos. Realizaram-se inicialmente as análises de dissolução da amostra e o exame direto por microscopias estereoscópica e óptica. Constatou-se a presença de borra no gargalo e de sedimento com cristais diversos no fundo das garrafas. Posteriormente, os cristais foram encaminhados ao Departamento de Biologia Celular e Molecular da FMRP-USP para quantificar e identificar a composição através do microscópio eletrônico de varredura JeolJSM-6610LV e do EDS (Energy Dispersive X-ray Spectroscopy). A ausência de sílica confirmou a pesquisa negativa para caco de vidro, e a composição dos elementos encontrados sugeriu a presença de sais de tartarato de cálcio e potássio no vinho. Apesar destes cristais não serem prejudiciais à saúde, a alteração da aparência do produto pode causar rejeição pelo consumidor...