New B-type cyclin synthesis is required between meiosis I and II during Xenopus oocyte maturation

Progression through meiosis requires two waves of maturation promoting factor (MPF) activity corresponding to meiosis I and meiosis II. Frog oocytes contain a pool of inactive "pre-MPF" consisting of cyclin-dependent kinase 1 bound to B-type cyclins, of which we now find three previously unsuspected members, cyclins B3, B4 and B5. Protein synthesis is required to activate pre-MPF, and we show here that this does not require new B-type cyclin synthesis, probably because of a large maternal stockpile of cyclins B2 and B5. This stockpile is degraded after meiosis I and consequently, the activation of MPF for meiosis II requires new cyclin synthesis, principally of cyclins B1 and B4, whose translation is strongly activated after meiosis I. If this wave of new cyclin synthesis is ablated by antisense oligonucleotides, the oocytes degenerate and fail to form a second meiotic spindle. The effects on meiotic progression are even more severe when all new protein synthesis is blocked by cycloheximide added after meiosis I, but can be rescued by injection of indestructible B-type cyclins. B-type cyclins and MPF activity are required to maintain c-mos and MAP kinase activity during meiosis II, and to establish the metaphase arrest at the end of meiotic maturation. We discuss the interdependence of c-mos and MPF, and reveal an important role for translational control of cyclin synthesis between the two meiotic divisions.     

Estrogenic alkylphenols induce cell death by inhibiting testis endoplasmic reticulum Ca(2+) pumps

Industrial alkylphenols in the environment may act as "xenoestrogens" to disrupt testicular development and decrease male fertility. Amongst possible targets for these compounds are testicular Sertoli cells, which nurture the developing sperm cells. We demonstrate that SERCA 2 and 3 Ca(2+) pumps are relatively abundant in rat testis microsomal membranes, and also in Sertoli, myoid, and TM4 cells (a Sertoli cell line). A number of estrogenic alkylphenols such as nonylphenol, octylphenol, bisphenol A, and butylated hydroxytoluene all inhibit testicular Ca(2+) ATPase in the low micromolar concentration range. These agents also mobilize intracellular Ca(2+) in intact TM4 cells in a manner consistent with the inhibition of ER Ca(2+) pumps. Alkylphenols dramatically decrease the viability of TM4 cells, an effect that is reversed by either a caspase inhibitor or by BAPTA, and is therefore consistent with Ca(2+)-dependent cell death via apoptosis. We postulate that alkylphenols disrupt testicular development by inhibiting ER Ca(2+) pumps, thus disturbing testicular Ca(2+) homeostasis.     

栲树种群的年龄结构及其生长特征

为了了解栲树的更新方式和更新动态,研究了栲树的生长特征和种群年龄结构.结果表明：栲树种群的年龄结构呈“间歇型”,经历了两个死亡高峰,并存在一个长达30年的断层;栲树的生长受光照的影响,具有很强的可塑性;由于林下光照弱且在垂直空间上不存在差异,栲树生长5～8年后进入生长的第1个抑制期,其年高生长速度可小于0.1 m,并可维持10年;栲树生长的第1个抑制期的起始时间对应着种群第1个死亡高峰期的结束时间,而其结束时间对应着种群第2个死亡高峰期的起始时间,表明栲树生长特征是影响其种群年龄结构的关键因素.     

Growth limiting substrate affects antibiotic production and associated metabolic fluxes in Streptomyces clavuligerus

Clavulanic acid biosynthesis by Streptomyces clavuligerus was dependent on the identity of the growth rate limiting nutrient in chemostat bioreactor culture (D=0.05 h^–1). In phosphate-limited media, a specific production rate of 3.65 mg_clav g_biomass h^–1 was observed while N-limited media supported 0.32 mg_clav g_biomass h^–1. No production was observed in C-limited media. Metabolic flux analysis suggested that changing the nutrient limitation affected the availability of the C₅ precursor. Flux through anaplerotic metabolism was consistent with this, reflecting the lower rate of utilisation of 2-oxo-glutarate from the tricarboxylic acid (TCA) cycle for glutamate and, ultimately, C₅ precursor production, when antibiotic was not produced. We propose that C-limitation restricts the capacity for anaplerotic metabolism, minimising the potential for extensive TCA-cycle derived biosynthesis (the first stage in production of the C₅ precursor). N-Limitation would restrict the availability of nitrogen for amino acid biosynthesis (the next stage). Under P-limitation neither of these restrictions would apply.     

Model analysis of steady-state ventilatory response to CO2 into component factors

A method is described to partition measured values of steady-state ventilatory response into an estimation of the blood flow in the respiratory controller and the sensitivity of the controller to CO2 assuming proportional control. The analysis is derived from the describing equations of a computer model and leads to the definition of a grid of lines emanating from a hypothetical reference point at negative ventilation and zero central nervous system metabolism. Data from the literature reporting differences in CO2 response among normal subjects and changes in resting ventilation and cerebral blood flow with age are reinterpreted from this perspective. Use of a structural model to interpret physiological data is shown to give a different meaning to data reduction in contrast to interpretation using statistical models like regression.     

Translational regulation of protein synthesis, in response to light, at a critical stage of Volvox development

In Volvox cultures synchronized by a light-dark cycle, juveniles containing presumptive somatic and reproductive cells are produced during the dark, but their cells do not differentiate until after the lights come on. The pattern of protein synthesis changes rapidly after the lights come on. Action spectra and effects of photosynthesis inhibitors indicate that this protein synthetic change is not simply a consequence of renewed flow of energy from illuminated chloroplasts. Actinomycin, at a level adequate to block the response to heat shock, has virtually no effect on the response of the same cells to light; furthermore, RNAs isolated from unilluminated and illuminated juveniles yield indistinguishable in vitro translation products. We conclude, therefore, that this effect of light is exerted almost exclusively at the translational level, generating one of the most striking examples of translational regulation yet described.     

Normal human endometrium in cell culture

Summary Separation of human endometrium into its epithelial and stromal components has been achieved through collagenase digestion and has permitted a study of these two cell populations under specific experimental culture conditions. The stromal cell populations showed a progesterone response, were easily handled in culture, and displayed a limited in vitro life span typical of human diploid fibroblasts. In contrast, epithelium only survived in shortterm primary culture and showed no clear hormone response. High-density epithelial cultures remained viable for longer periods in culture. Comparisons between resurfacing endometrial epithelial cells in vivo and epithelial cells migrating from explants in vitro suggested that this initial epithelial migration in vitro was the counterpart of the repair response in vivo. We are much in debt to Dr. R. C. Hallowes (Department of Pathology, Imperial Cancer Research Fund) for his guidance and encouragement throughout the course of this work. We also gratefully acknowledge Dr. P. N. Riddle (Time-Lapse Cinematography Unit, Imperial Cancer Research Fund) for carrying out the time-lapse cinematography; Mrs. Lyn Rolph (Stereoscan Unit, Bedford College, University of London) for assisting with the SEM; and Mr. G. D. Leach for his competent help with the photography.     

Natural Selection Reduced Diversity on Human Y Chromosomes

The human Y chromosome exhibits surprisingly low levels of genetic diversity. This could result from neutral processes if the effective population size of males is reduced relative to females due to a higher variance in the number of offspring from males than from females. Alternatively, selection acting on new mutations, and affecting linked neutral sites, could reduce variability on the Y chromosome. Here, using genome-wide analyses of X, Y, autosomal and mitochondrial DNA, in combination with extensive population genetic simulations, we show that low observed Y chromosome variability is not consistent with a purely neutral model. Instead, we show that models of purifying selection are consistent with observed Y diversity. Further, the number of sites estimated to be under purifying selection greatly exceeds the number of Y-linked coding sites, suggesting the importance of the highly repetitive ampliconic regions. While we show that purifying selection removing deleterious mutations can explain the low diversity on the Y chromosome, we cannot exclude the possibility that positive selection acting on beneficial mutations could have also reduced diversity in linked neutral regions, and may have contributed to lowering human Y chromosome diversity. Because the functional significance of the ampliconic regions is poorly understood, our findings should motivate future research in this area.     

A second form of adenine phosphoribosyltransferase in Arabidopsis thaliana with relative specificity towards cytokinins

Adenine phosphoribosyltransferase (APRTase) is an important enzyme for its ability to convert adenine, a by-product of many biochemical reactions, into AMP. By functional complementation of an Escherichia coli mutant, cDNAs encoding two APRTases have been cloned from Arabidopsis thaliana. One of the cDNAs (ATapt1)has been previously identified while the second (ATapt2) is of a previously unknown type. Kinetic analysis of the two enzymes purified from E. coli expressing the two cDNAs indicates that ATapt2 has a higher affinity for cytokinin than the ATapt1. RNase protection studies indicate that the ATapt2, is not expressed in leaves. Analysis of the gene structure indicates that ATapt2 has identical intron positions to ATapt1, but neither the intron sequence nor intron size are conserved between the two genes. The implications of a second, differentially expressed APRTase with affinity for both adenine and cytokinin are discussed.