Cell cycle parameters of slowly growing Escherichia coli B/r studied by flow cytometry

The cell cycle kinetics of Escherichia coli B/r A and B/r K cells were studied by flow cytometry. Three-dimensional histograms of cell cultures show the number of cells as a function of cellular DNA and protein contents and give detailed pictures of the cell cycle distribution with regard to these parameters. Histograms of slowly growing chemostat cultures showed that cell cycle periods B and C + D increase with a decreasing growth rate and that the B period occupies an increasing fraction of the cycle. The DNA replication patterns of B/r A and K were found to be quite similar. At extremely low growth rates (doubling time [T] = 17 h), B/r A cells had a B period of 0.8 T, a C period of 0.1 T, and a D period of 0.1 T, and B/r K cells (T = 16 h) had a B period of 0.6 T, a C period of 0.15 T, and a D period of 0.25 T. Mass increase, i.e., essentially protein synthesis, was seen in all three periods of the cell cycle. For B/r A cells, the average rate of mass increase was 11 times greater in the D period than in the B period, whereas for B/r K cells the rate of mass increase was twice as great in the D period as in the B period. The DNA and cell size distributions of batch cultures in exponential growth were found to vary with time, indicating that such cultures are not suitable for studies of cell cycle kinetics.     

The mouse pubic symphysis as a remodeling system: morphometrical analysis of proliferation and cell death during pregnancy, partus and postpartum

Marked changes in mice pubic symphysis occur by the end of pregnancy. Tissue remodeling involves a dynamic balance between cell proliferation and programmed cell death as well as changes in the extracellular matrix components. Therefore, it is important to consider both of these cellular behaviors when investigating the mechanism that regulates interpubic tissue remodeling, growth during late pregnancy and partus ensuring involution during the postpartum period. Proliferating and programmed death cells were identified by immunohistochemistry (proliferating cell nuclear antigen and TUNEL detection, respectively) and the rates at which these processes occurred were determined by morphometric analysis. The results demonstrated that cellular proliferation was intense during the period of ligament formation, from D15 to D18, thereafter abruptly declining on D19. From parturition (D19) onwards, an ever-increasing decline in the cellular proliferation levels could be observed. The quantitative analyses of cellular death showed opposite results when compared to cellular proliferation. During early pregnancy the cycle of cellular renovation was clearly proliferative and during late mouse pregnancy the cycle was directed by programmed cellular death. Although the high levels of cellular death during postpartum involution could be shown by the TUNEL-positive cells, we were unable to observed picnotic nucleus at the light microscopy.     

Identification of a novel response regulator required for the swarmer-to-stalked-cell transition in Caulobacter crescentus.

The onset of motility late in the Caulobacter crescentus cell cycle depends on a signal transduction pathway mediated by the histidine kinase PleC and response regulator DivK. We now show that pleD, whose function is required for the subsequent loss of motility and stalk formation by the motile swarmer cell, encodes a 454-residue protein with tandem N-terminal response regulator domains D1 and D2 and a novel C-terminal GGDEF domain. The identification of pleD301, a semidominant suppressor of the pleC Mot phenotype, as a mutation predicted to result in a D-53-->G change in the D1 domain supports a role for phosphorylation in the PleD regulator. Disruptions constructed in the pleD open reading frame demonstrated that the gene is not essential and that the pleC phenotype can also be suppressed by a recessive, loss-of-function mutation. These results suggest that PleD is part of a signal transduction pathway controlling stalked-cell differentiation early in the C. crescentus cell cycle.     

DIEL VARIATIONS OF CARBOHYDRATES AND NEUTRAL LIPIDS IN NITROGEN‐SUFFICIENT AND NITROGEN‐STARVED CYCLOSTAT CULTURES OF ISOCHRYSIS SP.1

The goal of this study was to investigate the time response of two major carbon (C) reserves, respectively neutral lipids (NL) and total carbohydrate (TC), in the Haptophyte Isochrysis sp. growing in nitrogen (N)‐sufficient or N‐starved conditions and under light:dark (L:D) cycles. Experiments were carried out in a cyclostat culture system that allowed the following of the dynamics of the main cell compounds at both hourly and daily time scales. Under N‐sufficient conditions, the L:D cycles cause the population to be synchronized, with most of the cells dividing at the beginning of the dark period. The C‐specific growth rate was maximal around midday and negative during the dark period due to respiration processes. NL and TC both accumulated during the day and consumed during the night. We showed that NL and TC are highly dynamic compounds, as more than three quarters of NL and TC accumulated during the light period were consumed during the dark period. In contrast to NL, phospholipid and glycolipid to C ratios remained quite stable during the light/dark cycles. The major effect of N starvation on the NL and TC dynamics was to uncouple their diel variations from the L:D cycle, in two different ways depending on their respective role during short‐term acclimation. Whereas the TC per cell ratio increased rapidly to reach a stable value in response to N starvation, NL per cell continued to oscillate, but with a pattern out of phase with the L:D cycle.     

Effect of Changing the Light/Dark Schedule, the Time of Onset of the Light or Dark Period, or the Daylength, on Rhythms of Epidermal Cell Proliferation

Rhythms of labeling and mitotic indices were studied in the hindlimb epidermis of the anuran tadpole Rana pipiens under different light/dark (LD) cycles and daylengths in order to examine the role of the various parameters of the lighting regimen in setting the periods of the rhythms and the timing of the cell proliferation peaks. Altering the time of, or inverting, the 12 h light period on a 24 h day resulted in phase shifting of basically bimodal circadian rhythms with peaks in the light and dark. Thus the cell proliferation rhythms were entrained to the LD cycle. These rhythms also entrained to noncircadian schedules since they lengthened on a 15L : 15D cycle and shortened on a 9L : 9D cycle, although the bimodal characteristic of a peak in the light and a peak in the dark remained. Studies of 18L: 6D and 6L : 18D cycles in which either the time of onset of light or dark was changed relative to the 12L: 12D control indicated that the onset of dark may regulate the timing of the labeling index peaks while the onset of light may determine the time of occurrence of mitotic index peaks. Control of the timing of labeling and mitotic index peaks by different parameters of the LD cycle suggests a mechanism for cell cycle regulation by the environmental lighting schedule. Analysis of the rhythms on all the cycles studied suggested that labeling index rhythms equal the length of, or twice the length of, the dark period. Mitotic index rhythms equal the daylfength or a multiple of the length of the dark period.     

The RAG proteins in V(D)J recombination: more than just a nuclease

V(D)J recombination is the process that generates the diversity among T cell receptors and is one of three mechanisms that contribute to the diversity of antibodies in the vertebrate immune system. The mechanism requires precise cutting of the DNA at segment boundaries followed by rejoining of particular pairs of the resulting termini. The imprecision of aspects of the joining reaction contributes significantly to increasing the variability of the resulting functional genes. Signal sequences target DNA recombination and must participate in a highly ordered protein-DNA complex in order to limit recombination to appropriate partners. Two proteins, RAG1 and RAG2, together form the nuclease that cleaves the DNA at the border of the signal sequences. Additional roles of these proteins in organizing the reaction complex for subsequent steps are explored.     

INTERACTIONS OF LIGHT/DARK CYCLES AND NITROGEN PULSES ON THE TIMING OF CELL DIVISION IN THE NITROGEN-LIMITED MARINE DIATOM CYLINDROTHECA FUSIFORMIS (BACILLARIOPHYCEAE)1

Cell division in most eukaryotic algae grown on alternating periods of light and dark (LD) is synchronized or phased so that cell division occurs only during a restricted portion of the LD cycle. However, the phase angle of the cell division gate, the time of division relative to the beginning of the light period, is known to be affected by growth conditions such as nutrient status and temperature. In this study, it is shown that the phase angle of cell division in a diatom, Cylindrotheca fusiformis Reimann and Lewin, is affected by the N-limited growth rate; cell division occurred later in the dark period (12:12 h LD cycle) when the growth rate was infradian (D = 0.42 d^?1) than when it was ultradian (D = 1.0 d^?1). Nitrogen-pulses did not affect the phase angle of the division gate, but could shift the time of peak cell division activity within the division gate. The effects, if any, of N-pulses were dependent upon the growth rate and the time of day that the pulses were administered. These responses indicate that the timing of cell division in this diatom is not determined solely by the zeitgeber from the LD cycle, but rather that a LD cycle control mechanism and a N-mediated control mechanism are both involved and are somewhat interdependent. In addition, an increase in protein was observed immediately after administering a N-pulse to C. fusiformis in the ultradian growth mode indicating that the accumulation of protein can be uncoupled from the cell division cycle.     

DNA ligase IV mutations identified in patients exhibiting developmental delay and immunodeficiency.

DNA ligase IV functions in DNA nonhomologous end-joining and V(D)J recombination. Four patients with features including immunodeficiency and developmental and growth delay were found to have mutations in the gene encoding DNA ligase IV (LIG4). Their clinical phenotype closely resembles the DNA damage response disorder, Nijmegen breakage syndrome (NBS). Some of the mutations identified in the patients directly disrupt the ligase domain while others impair the interaction between DNA ligase IV and Xrcc-4. Cell lines from the patients show pronounced radiosensitivity. Unlike NBS cell lines, they show normal cell cycle checkpoint responses but impaired DNA double-strand break rejoining. An unexpected V(D)J recombination phenotype is observed involving a small decrease in rejoining frequency coupled with elevated imprecision at signal junctions.     

Ras is active throughout the cell cycle,but is able to induce cyclin D1 only during G2 phase

The control of cell cycle progression has been studied in asynchronous cultures using image analysis and time lapse techniques. This approach allows determination of the cycle phase and signaling properties of individual cells, and avoids the need for synchronization. In past studies this approach demonstrated that continuous cell cycle progression requires the induction of cyclin D1 levels by Ras, and that this induction takes place during G2 phase. These studies were designed to understand how Ras could induce cyclin D1 levels only during G2 phase. First, in studies with a Ras-specific promoter and cellular migration we find that endogenous Ras is active in all cell cycle phases of actively cycling NIH3T3 cells. This suggests that cyclin D1 induction during G2 phase is not the result of Ras activation specifically during this cell cycle period. To confirm this suggestion oncogenic Ras, which is expected to be active in all cell cycle phases, was microinjected into asynchronous cells. The injected protein induced cyclin D1 levels rapidly, but only in G2 phase cells. We conclude that in the continuously cycling cell the targets of Ras activity are controlled by cell cycle phase, and that this phenomenon is vital to cell cycle progression.     

D609 对Neuro-2a 脑神经瘤细胞增殖和细胞周期的影响

目的:探讨小分子化合物D609对脑神经瘤细胞Neuro-2a的生长抑制及诱导细胞周期阻滞的效应,并初步研究其机制。方法:采用CCK-8法检测D609对Neuro-2a细胞的生长抑制作用;利用流式细胞术(FACS)检测D609处理对细胞周期进程的影响;利用免疫印迹实验(Western blot)检测不同浓度的D609处理后,细胞裂解液中细胞周期蛋白抑制因子p27的表达水平。结果:CCK-8的实验结果显示,加入150μmol/L D609处理72小时后,细胞生长受到明显地抑制,且伴有剂量依赖效应;流式细胞术的结果表明,D609处理使细胞周期阻滞在G0/G1期;免疫印迹的结果表明药物处理提升了p27的表达,且随药物浓度升高其表达亦增强。结论:D609可以有效地抑制Neuro-2a细胞的生长;进一步研究表明药物处理可以提升p27的表达水平并可以诱导将细胞阻滞在G0/G1期。因此,此研究将为脑神经瘤的治疗提供借鉴。     

Exploring temporal trend of morphological variability of a dominant diatom in response to environmental factors in a large subtropical river

The temporal trend of morphological variability of Aulacoseira granulata (Ehrenberg) Simonsen and its response to environments was studied within the downstream region of a large subtropical river, the Pearl River (China), through time-series sampling during 2009. Both wavelet analysis and redundancy analysis (RDA) were used to demonstrate not only the correlations between morphological parameters but also the correlations between morphology and environments. High coherence between morphological parameters, especially cell size, was confirmed; but the coherence, especially that between cell and filament, could easily be impacted by water turbulence which was associated with discharge, this might reflect the interaction between life cycle and size selectivity. Moreover, phase angles in wavelet figures illustrated that cell diameter was the most sensitive parameter to environmental variations, and changes in cell diameter triggered the size change in both cell and filament; thus through this way cell and filament size variations could be related.Based on the annual variation pattern of morphological parameters, the morphology of A. granulata exhibited an integrated cycle; during which morphological parameters might have different responses to physicochemical factors. Water temperature was closely associated with algal occurrence rates and size values during the spring–winter period. Algal life cycle could be affected by discharge, as well as filament length by allowing for selection of chains with optimum buoyancy. The responses of algal sizes to nutrients, especially silicate, phosphate, and total nitrogen, were associated with the start and end of a life cycle. These correlations between size and nutrients were supported by both wavelet analysis and RDA. Moreover, the extremely high values at the end of the year were explained as algal recruitment from benthos.     

Cyclin D2 ArrestsXenopusEarly Embryonic Cell Cycles

Xenopuscyclin D2 mRNA is a member of the class of maternal RNAs. It is rare and stable during early embryonic development. To investigate the potential role of cyclin D2 during early embryonic cell cycles, cyclin D2 was injected into one blastomere of a two-cell embryo. This injection induced a cell cycle arrest in the injected blastomere. To analyze more precisely the mechanism of this arrest, we took advantage of cycling egg extracts that recapitulate major events of the cell cycle when supplemented with demembranated sperm heads. WhenXenopuscyclin D2 is added to egg extracts, the first round of DNA replication occurs as in control extracts. However,Xenopuscyclin D2 blocks subsequent rounds of DNA replication and the oscillations of histone H1 kinase activity associated with cdc2 kinase, indicating that the cell cycle is arrested after the first S-phase. The block induced byXenopuscyclin D2 is not due to a lack of the mitotic cyclin B2 that accumulates normally. RadiolabeledXenopuscyclin D2 enters nuclei after completion of the first S-phase and remains stable over the entire period of the arrest. These features suggest thatXenopuscyclin D2 could play an original role during early development, controlling the G2-phase and/or the G2/M transition.     

A finite representation model for an asynchronous culture of E. coli

A computer model is described which models an asynchronous population of E. coli by using a large, but finite number of representative single cells. Asynchrony generation and maintenance occurs at the single cell level by modulating the activity of an enzyme responsible for septum formation. Such modulation introduces cycle time imprecision and does not require the introduction of any new parameters into the single-cell model. Based on comparisons to experiment, reasonable predictions are possible for changes of cellular dry weight during exponential growth and turbidostat washout, and overall chemostat cell yields and changes in cell number, glucose concentration, and cell size distribution for a chemostat subject to a step change in dilution rate. Additionally, a correlation between cell RNA content and size is predicted as is an inertial effect when chemostat residence time is decreased under conditions of initially high glucose concentrations. Limitations imposed by the model's finite nature and their solutions are discussed.     

Femininity and masculinity across the menstrual cycle: a relation to mate value

Numerous studies have shown that menstrual cycle related variations in sex hormones influence various cognitive processes. These shifts are considered as the evidence for a hormone-mediated adaptive design underlying human mating motivation. In a series of related studies we have shown that (i) femininity does not vary across the menstrual cycle, whereas masculinity is the most pronounced during the fertile period, (ii) masculinity, but not femininity, predicts shifts in spatial cognition across the menstrual cycle, and (iii) women with different positions on masculinity and femininity dimensions differ in their self-perceived mate value. These results suggest that (i) there might be a hormone mediated psychological mechanism making a woman more assertive and dominant during a short time-window when the conception is likely, (ii) menstrual cycle related shifts in cognitive abilities and mating motivation might have a common hormonal mechanism, and (iii) women's mate value (and indirectly her reproductive success) depends upon both feminine and masculine traits.     

Circadian activity rhythms in the crayfish Paranephrops zealandicus (Crustacea)

The diel activity rhythm shown by Paranephrops is usually unimodal with most activity occurring at night. In constant darkness, locomotor activity is expressed for up to 60 days as a free-running circadian rhythm. The period varies from 18 to 38 hours following a seasonal cycle (summer mean period 24 hours, winter mean 30 hours). No adaptive significance can be attached to either seasonal or individual variations in period and it is suggested that such variations are a consequence of the way in which the clock mechanism operates, the period being linked to the overall activity level following Aschoff's ( 1960 ) "Circadian Rule". Activity levels will inevitably vary individually and seasonally.