Rat vas deferens SERCA2 is modulated by Ca2+/calmodulin protein kinase II-mediated phosphorylation

Ca2+ pumps are important players in smooth muscle contraction. Nevertheless, little information is available about these pumps in the vas deferens. We have determined which subtype of sarco(endo)plasmic reticulum Ca2+-ATPase isoform (SERCA) is expressed in rat vas deferens (RVD) and its modulation by calmodulin (CaM)-dependent mechanisms. The thapsigargin-sensitive Ca2+-ATPase from a membrane fraction containing the highest SERCA levels in the RVD homogenate has the same molecular mass (∼115 kDa) as that of SERCA2 from the rat cerebellum. It has a very high affinity for Ca2+ (Ca0.5 = 780 nM) and a low sensitivity to vanadate (IC50 = 41 µM). These facts indicate that SERCA2 is present in the RVD. Immunoblotting for CaM and Ca2+/calmodulin-dependent protein kinase II (CaMKII) showed the expression of these two regulatory proteins. Ca2+ and CaM increased serine-phosphorylated residues of the 115-kDa protein, indicating the involvement of CaMKII in the regulatory phosphorylation of SERCA2. Phosphorylation is accompanied by an 8-fold increase of thapsigargin-sensitive Ca2+ accumulation in the lumen of vesicles derived from these membranes. These data establish that SERCA2 in the RVD is modulated by Ca2+ and CaM, possibly via CaMKII, in a process that results in stimulation of Ca2+ pumping activity.

Effect of estradiol and bisphenol A on human hepatoblastoma cell viability and telomerase activity

Sex hormones from environmental and physiological sources might play a major role in the pathogenesis of hepatoblastoma in children. This study investigated the effects of estradiol and bisphenol A on the proliferation and telomerase activity of human hepatoblastoma HepG2 cells. The cells were divided into 6 treatment groups: control, bisphenol A, estradiol, anti-estrogen ICI 182,780 (hereinafter ICI), bisphenol A+ICI, and estradiol+ICI. Cell proliferation was measured based on average absorbance using the Cell Counting-8 assay. The cell cycle distribution and apoptotic index were determined by flow cytometry. Telomerase activity was detected by polymerase chain reaction and a telomeric repeat amplification protocol assay. A higher cell density was observed in bisphenol A (P<0.01) and estradiol (P<0.05) groups compared with the control group. Cell numbers in S and G2/M phases after treatment for 48 h were higher (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h (P<0.05) were higher in these groups than in the control group. The cell density was also higher in bisphenol A+ICI (P<0.01) and estradiol+ICI (P<0.05) groups compared with the ICI group. Furthermore, cell numbers were increased in S and G2/M phases (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h were higher (P<0.05) in these groups than in the ICI group. Therefore, bisphenol A and estradiol promote HepG2 cell proliferation in vitro by inhibition of apoptosis and stimulation of telomerase activity via an estrogen receptor-dependent pathway.

Evaluation of energy expenditure in forward and backward movements performed by soccer referees

The aim of this study was to measure the energy expenditure for locomotor activities usually performed by soccer referees during a match (walking, jogging, and running) under laboratory conditions, and to compare forward with backward movements. The sample was composed by 10 male soccer referees, age 29±7.8 years, body mass 77.5±6.2 kg, stature 1.78±0.07 m and professional experience of 7.33±4.92 years. Referees were evaluated on two separate occasions. On the first day, maximal oxygen uptake (VO2max) was determined by a maximal treadmill test, and on the second day, the oxygen consumption was determined in different speeds of forward and backward movements. The mean VO2max was 41.20±3.60 mL·kg-1·min-1 and the mean heart rate achieved in the last stage of the test was 190.5±7.9 bpm. When results of forward and backward movements were compared at 1.62 m/s (walking speed), we found significant differences in VO2, in metabolic equivalents, and in kcal. However, the same parameters in forward and backward movements at jogging velocities (2.46 m/s) were not significantly different, showing that these motor activities have similar intensity. Backward movements at velocities equivalent to walking and jogging are moderate-intensity activities, with energy expenditure less than 9 kcal. Energy expenditure was overestimated by at least 35% when calculated by mathematical equations. In summary, we observed that backward movements are not high-intensity activities as has been commonly reported, and when calculated using equations available in the literature, energy expenditure was overestimated compared to the values obtained by indirect calorimetry.

CARACTERÍSTICAS DE QUALIDADE DO CONTRAFILÉ (m. L. dorsi) DE TOUROS JOVENS DA RAÇA NELORE

Com o objetivo de analisar as características de qualidade da carne de touros jovens da raça Nelore e os possíveis efeitos da idade, entre os 690 e os 780 dias, nessas características, efetuou-se o abate de 113 bovinos, que haviam sido confinados por um período de 109 dias com ração à base de 20% de concentrado e 80% de volumoso. Todas as carcaças foram tipificadas pela Inspeção Federal na sala de matança do frigorífico e resfriadas por 24 h em câmara fria (Tinicial=5°C; Tfinal=2°C). Meias carcaças (lado direito, N=51), do tipo B - do Sistema B R A S I L - de animais de 23 a 26 meses foram desossadas. Dois bifes (2,5cm de espessura) de contrafilé (m. L. dorsi) de cada uma foram embalados à vácuo e maturados por 7 dias a 0-2°C. O pH variou entre 5,44 e 5,83 e apenas duas amostras tinham pH ³ 5,70. O valor L* (luminosidade) médio foi de 34,85. As médias de umidade e gordura foram de 75,65% e 1,71%, respectivamente. A média de força de cisalhamento foi de 6,70kg e não foi influenciada pela idade do animal entre 690 e 734 dias, mas apresentou uma tendência (Teste t, p=0,22) de aumento entre 735 e 780 dias. Não se detectou influência da idade do animal nas demais características de qualidade analisadas (Teste t, p>0,20). Concluiu-se que o contrafilé de animais como esses pode ter problemas de aceitação na faixa mais exigente de mercado, e que carcaças tipificadas como B, presumivelmente o melhor tipo, não apresentam necessariamente as melhores características de qualidade da carne.

Infecção do trato urinário de repetição por fístula enterovesical secundária a doença diverticular do cólon: um relato de caso

INTRODUÇÃO: As fístulas enterovesicais (FEV) são comunicações patológicas entre a bexiga e as alças intestinais pélvicas. Trata-se de uma rara complicação decorrente de doenças inflamatórias e neoplásicas da pelve, além de casos resultantes de iatrogenia, e associa-se a altos índices de morbimortalidade. RELATO DO CASO: Trata-se de um paciente de 61 anos com um quadro de dor e distensão abdominal, vômitos, parada de eliminação de fezes e flatos. APP: Hipertenso, diabético, com antecedentes de disfunção vesical e infecções do trato urinário de repetição (ITUr) nos últimos três anos. Por meio da realização de ressonância magnética de abdômen e pelve, diagnosticou-se FEV associada à doença diverticular (DDC) do sigmoide. A conduta estabelecida consistiu em colectomia parcial com rebaixamento de colo e cistectomia parcial com colocação cirúrgica de cateter duplo jota à esquerda. DISCUSSÃO: Embora consista de afecção primária do trato digestivo, normalmente o paciente com DDC associada a FEV procura atendimento médico em decorrência de queixas do trato urinário. Nesse caso, a demora no diagnóstico fez com que a queixa principal fosse do trato digestivo e com antecedentes de queixas urinárias. CONCLUSÃO: Apesar de pouco frequente, a ocorrência de ITUr associada à DDC deve ser sempre considerada no diagnóstico diferencial das ITUr pela alta morbimortalidade.