Onset Of Quiescence Research Articles

Deciphering Solar Magnetic Activity: The Solar Cycle Clock

The Sun’s variability is controlled by the progression and interaction of the magnetized systems that form the 22-year magnetic activity cycle (the “Hale Cycle”) as they march from their origin at ∼55° latitude to the equator, over ∼19 years. We will discuss the end point of that progression, dubbed “terminator” events, and our means of diagnosing them. In this paper we expand on the Extended Solar Cycle framework to construct a new solar activity “clock” which maps all solar magnetic activity onto a single normalized epoch based on the terminations of Hale Magnetic Cycles. Defining phase 0*2π on this clock as the Terminators, then solar polar field reversals occur at ∼ 0.2*2π, and the geomagnetically quiet intervals centered around solar minimum start at ∼ 0.6*2π and end at the terminator, thus lasting 40% of the cycle length. At this onset of quiescence, dubbed a “pre-terminator,” the Sun shows a radical reduction in active region complexity and, like the terminator events, is associated with the time when the solar radio flux crosses F10.7 = 90 sfu. We use the terminator-based clock to illustrate a range of phenomena that further emphasize the strong interaction of the global-scale magnetic systems of the Hale Cycle: the vast majority, 96%, of all X-flares happen between the Terminator and pre-Terminator. In addition to the X-rays from violent flares, rapid changes in the number of energetic photons—EUV spectral emission from a hot corona and the F10.7 solar radio flux—impinging on the atmosphere are predictable from the Terminator-normalized unit cycle, which has implications for improving the fidelity of atmospheric modelling.

Onset of Quiescence Following p53 Mediated Down-Regulation of H2AX in Normal Cells

Normal cells, both in vivo and in vitro, become quiescent after serial cell proliferation. During this process, cells can develop immortality with genomic instability, although the mechanisms by which this is regulated are unclear. Here, we show that a growth-arrested cellular status is produced by the down-regulation of histone H2AX in normal cells. Normal mouse embryonic fibroblast cells preserve an H2AX diminished quiescent status through p53 regulation and stable-diploidy maintenance. However, such quiescence is abrogated under continuous growth stimulation, inducing DNA replication stress. Because DNA replication stress-associated lesions are cryptogenic and capable of mediating chromosome-bridge formation and cytokinesis failure, this results in tetraploidization. Arf/p53 module-mutation is induced during tetraploidization with the resulting H2AX recovery and immortality acquisition. Thus, although cellular homeostasis is preserved under quiescence with stable diploidy, tetraploidization induced under growth stimulation disrupts the homeostasis and triggers immortality acquisition.

Cell cycle related modulations in Runx2 protein levels are independent of lymphocyte enhancer-binding factor 1 (Lef1) in proliferating osteoblasts

Runt-related transcription factor Runx2 regulates osteogenic phenotype commitment and attenuates osteoblast growth. Runx2 levels are cell cycle regulated and maximal in the G1 phase of proliferating osteoblasts and during quiescence. The Wnt/Lrp5-Frizzled/beta-catenin/Lef-Tcf signaling cascade also controls progression along the osteogenic lineage with a net anabolic effect that promotes bone formation. However, Lef1 opposes the osteoblast maturation promoting activity of Runx2. Here we examined whether Lef1 controls Runx2 expression during the cell cycle or onset of quiescence in osteoblasts. We inhibited Lef1 expression using short hairpin (sh) RNA interference in stably transfected MC3T3-E1 cells. In asynchronously growing osteoblasts, expression of Lef1 shRNA diminishes Lef1 protein levels, but does not affect Runx2 levels. Cells arrested in different cell cycle stages using mimosine (late G1), hydroxyurea or aphidicolin (S phase) or nocodazole (mitosis) exhibit expected reductions in Runx2 protein levels despite reductions in Lef1. Serum deprived MC3T3-E1 cells normally upregulate Runx2 protein regardless of Lef1 deficiency, although loss of Lef1 reduces cyclin A and increases cyclin D1 expression upon serum withdrawal. Thus, Runx2 protein levels during the cell cycle and onset of quiescence are regulated independently of Lef1, one of the major transcriptional inducers of Wnt signaling in proliferating cells.

The replication capacity of intact mammalian nuclei in Xenopus egg extracts declines with quiescence, but the residual DNA synthesis is independent of Xenopus MCM proteins

In eukaryotes, the initiation of DNA synthesis requires the assembly of a pre-replicative complex (pre-RC) at origins of replication. This involves the sequential binding of ORC (origin-recognition-complex), Cdc6 and MCM proteins, a process referred to as licensing. After origin firing, the Cdc6 and MCM proteins dissociate from the chromatin, and do not rebind until after the completion of mitosis, thereby restricting replication to a single round in each cell cycle. Although nuclei normally become licensed for replication as they enter G(1), the extent to which the license is retained when cells enter the quiescent state (G(0)) is controversial. Here we show that the replication capacity of nuclei from Swiss 3T3 cells, in Xenopus egg extracts, is not lost abruptly with the onset of quiescence, but instead declines gradually. The decline in replication capacity, which affects both the number of nuclei induced to replicate and their subsequent rate of DNA synthesis, is accompanied by a fall in the level of chromatin-bound MCM2. When quiescent nuclei are incubated in egg extracts, they do not bind further MCMs unless the nuclei are first permeabilized. The residual replication capacity of intact nuclei must therefore be dependent on the remaining endogenous MCMs. Although high levels of Cdk activity are known to block MCM binding, we show that the failure of intact nuclei in egg extracts to increase their bound MCMs is not due to their uptake and accumulation of Cdk complexes. Instead, the failure of binding must be due to exclusion of some other binding factor from the nucleus, or to the presence within nuclei of an inhibitor of binding other than Cdk activity. In contrast to the situation in Xenopus egg extracts, following serum stimulation of intact quiescent cells, the level of bound MCMs does increase before the cells reach S phase, without any disruption of the nuclear envelope.

Significance test for seismicity rate changes before the 1987 Chiba-toho-oki earthquake (M 6.7) Japan

A precursory seismic quiescence lasting 1.5 ± 0.5 years was observed prior to the 1987 M 6.7 Chiba-toho-oki earthquake, Central Japan. This event was the largest mainshock to occur in the region in 60 years. A quantitative analysis of the seismicity rates, using two independent catalogs provided by the NIED and JMA networks, shows that the precursory seismic quiescence is centered in the shallower part of the rupture zone of the subsequent mainshock, at a depth of 20-40 km. At the hypocenter of the 1987 Chiba-toho-oki mainshock, a 50% increase in the seismicity rate was detected in the NIED data, coinciding in time with the onset of quiescence (1986.4 ± 0.5). The simultaneous appearance of both quiescence in the shallow part of the rupture zone observed in two catalogs, and a rate increase in the immediate hypocenter region, suggest that these phenomena are causally linked to the subsequent mainshock. However, a quantitative analysis of both catalogs reveals that the precursory quiescence and rate increase are not unique, since rate changes of this duration and significance often occur in the data. A rate change of this significance rating could probably not be detected as a precursor in a real time approach. For the aid of real time monitoring of seismicity rate changes, we introduce the method to calculate the 95-percentile of confidence level for the significant rate changes.

Effects of choline and quiescence on Drosophila choline acetyltransferase expression and acetylcholine production by transduced rat fibroblasts.

Rat-1 fibroblasts were transduced to express Drosophila choline acetyltransferase. The presence of an active enzyme in these cells (Rat-1/dChAT) was confirmed using various methods. Rat-1/dChAT fibroblasts released acetylcholine (ACh) into the culture medium. Moreover, intra- and extracellular levels of ACh could be increased by adding exogenous choline chloride. In addition, serum starvation or confluence-induced quiescence caused an 80% decrease in recombinant choline acetyltransferase activity (compared with actively growing cells). ACh release was also repressed in quiescent fibroblast cultures. Exogenous choline could mitigate the decrease in ACh release. These results indicate that Rat-1 fibroblasts can be genetically modified to produce ACh and that ACh release can be controlled by introducing choline into the culture medium. Furthermore, these data demonstrate that the expression of the retroviral promoter used in this study decreases with the onset of quiescence; however, exogenous choline can increase the amount of ACh released by quiescent fibroblasts.

Coordination of protein-DNA interactions in the promoters of human H4, H3, and H1 histone genes during the cell cycle, tumorigenesis, and development

Coordinate transcriptional control of replication-dependent human H4, H3, and H1 histone genes was studied by comparing levels of H3 and H1 histone promoter binding activities with those of H4 histone promoter factor HiNF-D during the cell cycle of both normal diploid and tumor-derived cells, as well as in fetal and adult mammalian tissues. Both H3 and H1 histone promoters interact with binding activities that, as with HiNF-D, are maximal during S-phase but at low levels in the G1-phase of normal diploid cells. However, these analogous DNA binding activities are constitutively maintained at high levels throughout the cell cycle in four different transformed and tumor-derived cells. Downregulation of the H3 and H1 histone promoter factors in conjunction with HiNF-D is observed in vivo at the onset of quiescence and differentiation during hepatic development. Hence, our results indicate a tight temporal coupling of three separate protein-DNA interactions in different histone promoters during the cell cycle, development, and tumorigenesis. This suggests that a key oscillatory, cell-growth-control mechanism modulates three analogous histone gene promoter protein-DNA interactions in concert. The derangement of this mechanism in four distinct tumor cells implies that concerted deregulation of these histone promoter factors is a common event resulting from heterogeneous aberrations in normal cell growth mechanisms during tumorigenesis. We postulate that this mechanism may be involved in the coordinate regulation of the human H4, H3, and H1 histone multigene families.

Transcriptional element H4-site II of cell cycle regulated human H4 histone genes is a multipartite protein/DNA interaction site for factors HiNF-D, HiNF-M, and HiNF-P: involvement of phosphorylation.

Cell cycle regulated gene expression was studied by analyzing protein/DNA interactions occurring at the H4-Site II transcriptional element of H4 histone genes using several approaches. We show that this key proximal promoter element interacts with at least three distinct sequence-specific DNA binding activities, designated HiNF-D, HiNF-M, and HiNF-P. HiNF-D binds to an extended series of nucleotides, whereas HiNF-M and HiNF-P recognize sequences internal to the HiNF-D binding domain. Gel retardation assays show that HiNF-D and HiNF-M each are represented by two distinct protein/DNA complexes involving the same DNA binding activity. These results suggest that these factors are subject to post-translational modifications. Dephosphorylation experiments in vitro suggest that both electrophoretic mobility and DNA binding activities of HiNF-D and HiNF-M are sensitive to phosphatase activity. We deduce that these factors may require a basal level of phosphorylation for sequence specific binding to H4-Site II and may represent phosphoproteins occurring in putative hyper- and hypo-phosphorylated forms. Based on dramatic fluctuations in the ratio of the two distinct HiNF-D species both during hepatic development and the cell cycle in normal diploid cells, we postulate that this modification of HiNF-D is related to the cell cycle. However, in several tumor-derived and transformed cell types the putative hyperphosphorylated form of HiNF-D is constitutively present. These data suggest that deregulation of a phosphatase-sensitive post-translational modification required for HiNF-D binding is a molecular event that reflects abrogation of a mechanism controlling cell proliferation. Thus, phosphorylation and dephosphorylation of histone promoter factors may provide a basis for modulation of protein/DNA interactions and H4 histone gene transcription during the cell cycle and at the onset of quiescence and differentiation.

Quiescence in 9L Cells and Correlation with Radiosensitivity and PLD Repair

The onset of quiescence, changes in X-ray sensitivity, and changes in capacity for potentially lethal damage (PLD) repair of unfed plateau-phase 9L44 cell cultures have been systematically investigated. The quiescent plateau phase in 9L cells was the result of nutrient deprivation and was not a cell contact effect. Eighty-five to 90% of the plateau-phase cells had a G1 DNA content and a growth fraction less than or equal to 0.15. The cell kinetic shifts in the population were temporally correlated with a developing radioresistance, which was characterized by a larger shoulder in the survival curve of the quiescent cells (Dq = 5.71 Gy) versus exponentially growing cells (Dq = 4.48 Gy). When the quiescent plateau-phase cells were refed, an increase in radiosensitivity resulted which approached that of exponentially growing 9L cells. Delayed plating experiments after irradiation of exponentially growing cells, quiescent plateau-phase cells, and synchronized early to mid-G1-phase cells indicated that while significant PLD repair was evident in all three populations, the quiescent 9L cells had a higher PLD repair capacity. Although data for immediate plating indicated that 9L cells may enter quiescence in the relatively radioresistant mid-G1 phase, the enhanced PLD repair capacity of quiescent cells cannot be explained by redistribution into G1 phase. When the unfed quiescent plateau-phase 9L cells were stimulated to reenter the cell cycle by replating into fresh medium, the first G1 was extended by 6 h compared with the G1 of exponentially growing or refed plateau-phase 9L cells.(ABSTRACT TRUNCATED AT 250 WORDS)

An experiment in earthquake prediction and the 7 May 1986 Andreanof Islands earthquake

Abstract The occurrence of the 7 May 1986 Andreanof Islands earthquake provides the opportunity to evaluate the earthquake forecast and specific prediction that had been formulated and promulgated, starting more than 2 yr before. The prediction experiment was based on the observation of seismic quiescence in the part of the Aleutian subduction zone monitored by the Central Aleutians (Adak) Seismic Network. The magnitude estimate ( M s 7 to 7.5) was based on the size of the quiescent zone and the seismic history of the Adak region since 1900. The time of occurrence was predicted as a test of the discovery by Habermann of a quiescence lasting about 3 yr prior to the 2 May 1971 Adak Canyon event, and the hypothesis that a characteristic precursor time is a valid concept. The place was originally forecast as “near Adak Island,” then specified with epicenter at the eastern margin of Adak Canyon, rupturing to the west under the canyon. The 7 May 1986 earthquake had magnitude M s 7.7 ( M w 8.0), with epicenter 140 km east of the predicted place, rupturing east of Adak Canyon and stopping in the area predicted as the nucleation point. The time of this event was 6 months later than predicted. Thus, the specifically predicted event has not happened. The largest part of the rupture did go through the segment that had been most active before the onset of quiescence and which experienced the sharpest and most persistent quiescence. A cause-and-effect relation between preparation for this earthquake and the quiescence is still to be firmly established, but the distribution of the pattern of quiescence relative to the epicentral location and the site of the greatest moment release suggest a physical connection between earthquake and seismicity patterns. The predictions of location and time were acknowledged throughout as highly speculative. No firm basis exists for quantitative estimation of the uncertainties to be assigned to the elements of an intermediate-term prediction. This experiment lends support to the validity of quiescence as a precursor, at least for some earthquakes, and demonstrates the need for local network data to provide the high resolution of seismicity required for prediction efforts based on seismicity patterns.

Thermodormancy in Seedlings of Eucalyptus obliqua L'herit

High (30°C) day and night temperatures reduced mainstem elongation in E. obliqua seedlings resulting in a temporary quiescence in height growth after 8 days of high temperature treatment; rapid stem elongation did not recommence when seedlings were returned to conditions usually favourable to growth. Associated with the onset of quiescence were: an increase in the respiratory quotient and a decrease in the respiration rate of upper terminal buds; an accumulation of inhibitory substances; and an increase in total polyphenols in the stem.

Pheromone Studies of the Two spotted Spider Mite. 2. Behavioral Response of Males to Quiescent Deutonymphs12

Adult male Tetranychus urticae Koch were attracted to quiescent female deutonymphs with the onset of quiescence. The consistency of male attraction varied from 40–85% during the early part of the period and gradually increased to 80–100% prior to adult emergence. Average duration of the quiescent period was 40.4 hours at 24°C and 23.4 hours at 30°C. Occasional males were found that did not respond consistently to quiescent deutonymphs. The cause was not determined. Male attraction was greater at 30°C and 20–30% relative humidity than at 30°C and 90–95% relative humidity. Trials at 24°C and 20–30% relative humidity were intermediate. It was concluded that tests for bioassay of T. urlicae female sex pheromone could be conducted at 30°C and 20–30% relative humidity but should include sufficient replications to measure erratic male behavior.