RESUMO
AIM: To investigate the effects of high magneto-gravitational environment on Ca(2+);]/calmodulin (CaM) signal of MG63 osteoblast-like cells. METHODS: A special designed large gradient high magnetic field could produce three different high magneto-gravitational environments including µg (12 T), 1 g (16 T) and 2 g (12 T). The effects of high magneto-gravitational environments on intracellular free Ca(2+);] concentration ([Ca(2+);](i);) and protein expression including calmodulin (CaM), myosin light chain kinases (MLCK) and phosphorylated Ca(2+);]/CaM dependent protein kinase II(pCaMKII) were measured by Fluo-3/AM or Western blot, respectively. RESULTS: When compared with control group, an increase of [Ca(2+);](i); of MG63 was caused by strong magnetic field; Compared to 2 g, µg decreased [Ca(2+);](i); of MG63. The protein expression of CaM and pCaMKIIof MG63 cells was decreased by simulated weightlessness. CONCLUSION: [Ca(2+);](i); of MG63 cells was increased by strong magnetic field; simulated weightlessness inhibited Ca(2+);/CaM signaling of MG63 cells.
Assuntos
Cálcio/metabolismo , Calmodulina/metabolismo , Gravitação , Fenômenos Magnéticos , Osteoblastos/metabolismo , Transdução de Sinais , Animais , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Linhagem Celular Tumoral , Ativação Enzimática , Regulação da Expressão Gênica , Humanos , Espaço Intracelular/metabolismoRESUMO
OBJECTIVE: Based on the sequence differences of Amelogenin homologous gene in the X and Y chromosomes, a pair of specific primers was designed to identify the sex of archaeological samples. METHODS: Ancient DNA fragments were extracted from the bones and teeth of sacrificial slaves with an improved method that combines phenol-chloroform extraction, silicon dioxide adsorption with ultrafiltration concentration. The polyacrylamide gel electrophoresis (PAGE) was used to detect PCR products. RESULTS: Seven in sixteen samples from eight graves showed positive results and the targeted segments were visible: a male with two bands of 106bp (Amel-X) and 112 bp (Amel-Y), while a female with only one band of 106 bp (Amel-X). Ancient DNA analyzing results from tooth samples are more marked than that from bones. CONCLUSION: The improved extraction method is more effective for ancient DNA extraction, which reduced the PCR inhibitors and lowered experimental costs. The sex determination technology based on Amelogenin homologous gene is an important and feasible method in the molecular archaeological research.
