Essential role of germinal vesicle material in the meiotic cell cycle of Xenopus oocytes

In almost all animal species, immature oocytes are arrested naturally in the first meiotic prophase, with a large nucleus called the germinal vesicle. A number of previous studies showed that both activation of maturation/M phase-promoting factor (MPF) (assayed by semiquantitative cytological methods) and some other maturational events occur essentially normally in enucleated oocytes from many amphibian species and mice. Hence, for nearly three decades, it has generally been believed that nuclear material is dispensable for MPF activation and the meiotic cell cycle in vertebrate oocytes. Here, we have challenged this view by examining the histone H1 kinase activities and the molecular forms of MPF in experimentally manipulated Xenopus oocytes. We show that oocytes injected with nuclear material undergo much more rapid MPF activation and maturation than uninjected control oocytes. Conversely, enucleated oocytes, unlike nucleated counterparts, undergo only weak MPF activation in meiosis I and no detectable MPF reactivation in meiosis II, the latter accompanying inhibitory tyrosine phosphorylation of cdc2 kinase, the catalytic subunit of MPF. These results argue strongly that nuclear material is indispensable for the meiotic cell cycle, particularly MPF reactivation (or cdc2 tyrosine dephosphorylation) on entry into meiosis II, in Xenopus oocytes. The classical and general view may thus need reconsideration.

Plo1 Kinase Recruitment to the Spindle Pole Body and Its Role in Cell Division in Schizosaccharomyces pombe

Polo kinases execute multiple roles during cell division. The fission yeast polo related kinase Plo1 is required to assemble the mitotic spindle, the prophase actin ring that predicts the site for cytokinesis and for septation after the completion of mitosis (Ohkura et al., 1995; Bahler et al., 1998). We show that Plo1 associates with the mitotic but not interphase spindle pole body (SPB). SPB association of Plo1 is the earliest fission yeast mitotic event recorded to date. SPB association is strong from mitotic commitment to early anaphase B, after which the Plo1 signal becomes very weak and finally disappears upon spindle breakdown. SPB association of Plo1 requires mitosis-promoting factor (MPF) activity, whereas its disassociation requires the activity of the anaphase-promoting complex. The stf1.1 mutation bypasses the usual requirement for the MPF activator Cdc25 (Hudson et al., 1990). Significantly, Plo1 associates inappropriately with the interphase SPB of stf1.1 cells. These data are consistent with the emerging theme from many systems that polo kinases participate in the regulation of MPF to determine the timing of commitment to mitosis and may indicate that pole association is a key aspect of Plo1 function. Plo1 does not associate with the SPB when septation is inappropriately driven by deregulation of the Spg1 pathway and remains SPB associated if septation occurs in the presence of a spindle. Thus, neither Plo1 recruitment to nor its departure from the SPB are required for septation; however, overexpression of plo1+ activates the Spg1 pathway and causes transient Cdc7 recruitment to the SPB and multiple rounds of septation.

An inhibitor of p34cdc2/cyclin B that regulates the G2/M transition in Xenopus extracts.

The activity of maturation-promoting factor (MPF), a protein kinase complex composed of p34cdc2 and cyclin B, is undetectable during interphase but rises abruptly at the G2/M transition to induce mitosis. After the synthesis of cyclin B, the suppression of MPF activity before mitosis has been attributed to the phosphorylation of p34cdc2 on sites (threonine-14 and tyrosine-15) that inhibit its catalytic activity. We previously showed that the activity of the mitotic p34cdc2/cyclin B complex is rapidly suppressed when added to interphase Xenopus extracts that lack endogenous cyclin B. Here we show that a mutant of p34cdc2 that cannot be inhibited by phosphorylation (threonine-14-->alanine, tyrosine-15-->phenylalanine) is also susceptible to inactivation, demonstrating that inhibitory mechanisms independent of threonine-14 and tyrosine-15 phosphorylation must exist. We have partially characterized this inhibitory pathway as one involving a reversible binding inhibitor of p34cdc2/cyclin B that is tightly associated with cell membranes. Kinetic analysis suggests that this inhibitor, in conjunction with the kinases that mediate the inhibitory phosphorylations on p34cdc2, maintains the interphase state in Xenopus; it may play an important role in the exact timing of the G2/M transition.

Conservadores ou progressistas: como o judiciário de Piracicaba/SP se posiciona nos pleitos pela proibição da queima da palha de cana-de-açúcar

Este estudo analisa os fatores, as tendências e as fundamentações jurídico-ideológicas que permeiam as decisões judiciais em Ações Civis Públicas, impetradas pelo Ministério Público do Estado de São Paulo (MPSP) e Federal (MPF), requerendo a proibição imediata da queima da palha de cana-de-açúcar, na região de Piracicaba/SP. Para isso, foram levantadas, junto aos órgãos jurisdicionais da cidade de Piracicaba, as Ações Civis Públicas impetradas em face de grandes usinas e agropecuárias dedicadas ao cultivo e industrialização da cana-de-açúcar, requerendo a proibição da queima da palha na lavoura desde o advento da Lei nº 7.347 de 1985 até 2014, ano em que finalizou-se a pesquisa empírica desta dissertação. Nesses padrões, a pesquisa arrojou a existência de quatro processos, dos quais foram extraídas para análise a totalidade de vinte-e-um (21) manifestações judiciais. O marco teórico-metodológico empregado foi a abordagem marxista da filosofia da linguagem, perspectiva capaz de proporcionar uma fecunda imersão não só no que se diz, mas nas condições de produção desse dizer, valorizando a função socioideológica da palavra, permitindo vislumbrar a visão social de mundo de quem a emprega ou do grupo social ao qual o autor pode ou não pertencer (VOLOCHINOV,1929; GOLDMANN, 1979). Conclui-se que, nos pleitos judiciais pela proibição imediata da queima da cana-de-açúcar em Piracicaba/SP, o Poder Judiciário se mostra majoritariamente conservador, arguindo e decidindo pela continuidade da prática em evidente defesa do setor sucroalcooleiro. No entanto, se reconhece a importância do tímido mas importante viés progressista por parte do judiciário, uma vez que somente nas contradições presentes nas diferentes práxis jurídicas pode avançar-se no processo permanente de construção de um direito comprometido com a obtenção de igualdade e justiça social.

Influência da paridade, do tipo de parto e do nível de atividade física sobre a musculatura do assoalho pélvico em mulheres na pós-menopausa

The post-menopause stage is characterized by hormonal and organic alterations of ovarian failure. One of the most important of these is muscles alterations of the pelvic floor (MPF). According to current literature, in young women gynecological and obstetric factors, as well as lifestyles and habits influence that loss of function. However, there is still uncertainty about the influence of those variables in the MPF functions in post-menopause women. Thus, this study aimed at seeing if there is an influence from number of births, the type of birth and the level of physical activity on the MPF of post-menopause women. Another objective of this study was to compare MPF force in women who had had vaginal births with those who had been subjected to cesarean sections, those with different levels of physical activity and those with artificial and natural menopause in the initial and latter stages. Furthermore, the test of muscular force was compared to perineometry. Using observational, analytical and transversal observations, 100 women in the post-menopausal stage of life, between the ages of 45 and 65, were examined. They were divided according to the menopausal stage into three groups: women who had undergone hysterectomies, those in the initial stages of postmenopause and those in the late stage of postmenpause. The patients were questioned about social, demographic, gynecological and obstetric factors. All the volunteers were submitted to a physical examination where their height and weight were measured to arrive at the corporal mass index and their waist measurements were taken. The evaluation of the pelvic floor was conducted with muscular force tests and perineometry. These results were analyzed with statistical description and ANOVA statistical tests, multiple regression and Kolmogorov-Smirnov evaluations. The results showed homogeneity with regard to social demographic and anthropometric characteristics among the women in the final test sample (n=85). It was also seen that most of the women in all three groups were married (p=0.51) and catholic (p=0.13). The average per capital income varied between $R585.47 (+/-466.67) and $R1,271.83 (+/-1,748.95), with no significant difference between the groups (p=0.05). The G>6 group presented an average age between 58.95 (+/-3.96) which was significantly greater that the G<6 group´s average age (53.21+/- 3.88) (p=0.000). There was no difference between the groups´ anthropometric characteristics of weight (p=0.32), height (p=0.72) and corporal mass index (p=0.34), nor in the waist measurements (p=0.33). Furthermore, no significant difference was noted in the MPF function of women who had had normal births, cesarean sections or a combination of the two (TFM p=0.897; perineum measurement p=0.502). Likewise, no differences were seen in the MPF function of women who had one, two to three or four or more births (TFM p=0.28, perineum measurement p=0.13). Finally, no difference was perceived among those with different levels of physical activity (TFM p=0.663; perineum measurement p=0.741). Therefore, we found that the type of delivery, number of births and physical activity had no influence on the muscular function of the pelvic floor among the women studied. It is believed that decline in muscular function in post-menopause women is fundamentally related to the process of aging.