From short pulses to short breaks : exotic plasma bullets via residual electron control

Plasma plumes with exotically segmented channel structure and plasma bullet propagation are produced in atmospheric plasma jets. This is achieved by tailoring interruptions of a continuous DC power supply over the time scales of lifetimes of residual electrons produced by the preceding discharge phase. These phenomena are explained by studying the plasma dynamics using nanosecond-precision imaging. One of the plumes is produced using 2-10μs interruptions in the 8kV DC voltage and features a still bright channel from which a propagating bullet detaches. A shorter interruption of 900ns produces a plume with the additional long conducting dark channel between the jet nozzle and the bright area. The bullet size, formation dynamics, and propagation speed and distance can be effectively controlled. This may lead to micrometer-and nanosecond-precision delivery of quantized plasma bits, warranted for next-generation health, materials, and device technologies.

Stop and revive? The effectiveness of nap and active rest breaks for reducing driver sleepiness

Objectives The purpose for this study was to determine the relative benefit of nap and active rest breaks for reducing driver sleepiness. Methods Participants were 20 healthy young adults (20-25 years), including 8 males and 12 females. A counterbalanced within-subjects design was used such that each participant completed both conditions on separate occasions, a week apart. The effects of the countermeasures were evaluated by established physiological (EEG theta and alpha absolute power), subjective (Karolinska Sleepiness Scale), and driving performance measures (Hazard Perception Task). Participants woke at 5am, and undertook a simulated driving task for two hours; each participant then had either a 15-minute nap opportunity or a 15-minute active rest break that included 10 minutes of brisk walking, followed by another hour of simulated driving. Results The nap break reduced EEG theta and alpha absolute power and eventually reduced subjective sleepiness levels. In contrast, the active rest break did not reduce EEG theta and alpha absolute power levels with the power levels eventually increasing. An immediate reduction of subjective sleepiness was observed, with subjective sleepiness increasing during the final hour of simulated driving. No difference was found between the two breaks for hazard perception performance. Conclusions Only the nap break produced a significant reduction in physiological sleepiness. The immediate reductions of subjective sleepiness following the active rest break could leave drivers with erroneous perceptions of their sleepiness, particularly as physiological sleepiness continued to increase after the break.

Double-strand breaks and the concept of short- and long-term epigenetic memory

Double-strand breaks represent an extremely cytolethal form of DNA damage and thus pose a serious threat to the preservation of genetic and epigenetic information. Though it is well-known that double-strand breaks such as those generated by ionising radiation are among the principal causative factors behind mutations, chromosomal aberrations, genetic instability and carcino-genesis, significantly less is known about the epigenetic consequences of double-strand break formation and repair for carcinogenesis. Double-strand break repair is a highly coordinated process that requires the unravelling of the compacted chromatin structure to facilitate repair machinery access and then restoration of the original undamaged chromatin state. Recent experimental findings have pointed to a potential mechanism for double-strand break-induced epigenetic silencing. This review will discuss some of the key epigenetic regulatory processes involved in double-strand break (DSB) repair and how incomplete or incorrect restoration of chromatin structure can leave a DSB-induced epigenetic memory of damage with potentially pathological repercussions

Evaluation of the efficacy of radiation-modifying compounds using γH2AX as a molecular marker of DNA double-strand breaks

Radiation therapy is a widely used therapeutic approach for cancer. To improve the efficacy of radiotherapy there is an intense interest in combining this modality with two broad classes of compounds, radiosensitizers and radioprotectors. These either enhance tumour-killing efficacy or mitigate damage to surrounding non-malignant tissue, respectively. Radiation exposure often results in the formation of DNA double-strand breaks, which are marked by the induction of H2AX phosphorylation to generate γH2AX. In addition to its essential role in DDR signalling and coordination of double-strand break repair, the ability to visualize and quantitate γH2AX foci using immunofluorescence microscopy techniques enables it to be exploited as an indicator of therapeutic efficacy in a range of cell types and tissues. This review will explore the emerging applicability of γH2AX as a marker for monitoring the effectiveness of radiation-modifying compounds.

Methylproamine protects against ionizing radiation by preventing DNA double-strand breaks

Purpose The majority of cancer patients will receive radiotherapy (RT), therefore, investigations into advances of this modality are important. Conventional RT dose intensities are limited by adverse responses in normal tissues and a primary goal is to ameliorate adverse normal tissue effects. The aim of these experiments is to further our understanding regarding the mechanism of radioprotection by the DNA minor groove binder, methylproamine, in a cellular context at the DNA level. Materials and methods We used immunocytochemical methods to measure the accumulation of phosphorylated H2AX (γH2AX) foci following ionizing radiation (IR) in patient-derived lymphoblastoid cells exposed to methylproamine. Furthermore, we performed pulsed field gel electrophoresis DNA damage and repair assays to directly interrogate the action of methylproamine on DNA in irradiated cells. Results We found that methylproamine-treated cells had fewer γH2AX foci after IR compared to untreated cells. Also, the presence of methylproamine decreased the amount of lower molecular weight DNA entering the gel as shown by the pulsed field gel electrophoresis assay. Conclusions These results suggest that methylproamine acts by preventing the formation of DNA double-strand breaks (dsbs) and support the hypothesis that radioprotection by methylproamine is mediated, at least in part, by decreasing initial DNA damage.

Effect of Temperature Variation on Sister Chromatid Exchange Frequency in Cultured Human Lymphocytes

The effect of temperature variation on sister chromatid exchange (SCE) frequencies in human lymphocytes was studied. An increase as well as decrease in incubation temperature of cells leads to a higher frequency of sister chromatid exchanges than in cultures grown at 37°C. In addition, it was observed that mitotic: index and cell cycle duration were affected by low temperature.

Interactions between rotation breaks, tillage and N management on sugarcane grown at Bundaberg and Ingham.

The impact of cropping histories (sugarcane, maize and soybean), tillage practices (conventional tillage and direct drill) and fertiliser N in the plant and 1st ratoon (1R) crops of sugarcane were examined in field trials at Bundaberg and Ingham. Average yields at Ingham (Q200) and Bundaberg (Q151) were quite similar in both the plant crop (83 t/ha and 80 t/ha, respectively) and the 1R (89 t/ha v 94 t/ha, respectively), with only minor treatment effects on CCS at each site. Cane yield responses to tillage, break history and N fertiliser varied significantly between sites. There was a 27% yield increase in the plant crop from the soybean fallow at Ingham, with soybeans producing a yield advantage over continuous cane, but there were no clear break effects at Bundaberg - possibly due to a complex of pathogenic nematodes that responded differently to soybeans and maize breaks. There was no carryover benefit of the soybean break into the 1R crop at Ingham, while at Bundaberg the maize break produced a 15% yield advantage over soybeans and continuous cane. The Ingham site recorded positive responses to N fertiliser addition in both the plant (20% yield increase) and 1R (34% yield increase) crops, but there was negligible carryover benefit from plant crop N in the 1R crop, or of a reduced N response after a soybean rotation. By contrast, the Bundaberg site showed no N response in any history in the plant crop, and only a small (5%) yield increase with N applied in the 1R crop. There was again no evidence of a reduced N response in the 1R crop after a soybean fallow. There were no significant effects of tillage on cane yields at either site, although there were some minor interactions between tillage, breaks and N management in the 1R crop at both sites. Crop N contents at Bundaberg were more than 3 times those recorded at Ingham in both the plant and 1R crops, with N concentrations in millable stalk at Ingham suggesting N deficiencies in all treatments. There was negligible additional N recovered in crop biomass from N fertiliser application or soybean residues at the Ingham site. There was additional N recovered in crop biomass in response to N fertiliser and soybean breaks at Bundaberg, but effects were small and fertiliser use efficiencies poor. Loss pathways could not be quantified, but denitrification or losses in runoff were the likely causes at Ingham while leaching predominated at Bundaberg. Results highlight the complexity involved in developing sustainable farming systems for contrasting soil types and climatic conditions. A better understanding of key sugarcane pathogens and their host range, as well as improved capacity to predict in-crop N mineralisation, will be key factors in future improvements to sugarcane farming systems.

Micro-breaks matter: A diary study on the effects of energy management strategies on occupational well-being

Organizational researchers and practitioners are increasingly interested in self-regulatory strategies employees can use at work to sustain or improve their occupational well-being. A recent cross-sectional study on energy management strategies suggested that many work-related strategies (e.g., setting a new goal) are positively related to occupational well-being, whereas many micro-breaks (e.g., listening to music) are negatively related to occupational well-being. We used a diary study design to take a closer look at the effects of these energy management strategies on fatigue and vitality. Based on conservation of resources theory, we hypothesized that both types of energy management strategies negatively predict fatigue and positively predict vitality. Employees (N = 124) responded to a baseline survey and to hourly surveys across one work day (6.7 times on average). Consistent with previous research, between-person differences in the use of work-related strategies were positively associated with between-person differences in vitality. However, results of multilevel analyses of the hourly diary data showed that only micro-breaks negatively predicted fatigue and positively predicted vitality. These findings suggest that taking micro-breaks during the work day may have short-term effects on occupational well-being, whereas using work-related strategies may have long-term effects.

Cytosines, but Not Purines, Determine Recombination Activating Gene (RAG)-induced Breaks on Heteroduplex DNA Structures

The sequence specificity of the recombination activating gene (RAG) complex during V(D)J recombination has been well studied. RAGs can also act as structure-specific nuclease; however, little is known about the mechanism of its action. Here, we show that in addition to DNA structure, sequence dictates the pattern and efficiency of RAG cleavage on altered DNA structures. Cytosine nucleotides are preferentially nicked by RAGs when present at single-stranded regions of heteroduplex DNA. Although unpaired thymine nucleotides are also nicked, the efficiency is many fold weaker. Induction of single- or double-strand breaks by RAGs depends on the position of cytosines and whether it is present on one or both of the strands. Interestingly, RAGs are unable to induce breaks when adenine or guanine nucleotides are present at single-strand regions. The nucleotide present immediately next to the bubble sequence could also affect RAG cleavage. Hence, we propose “C(d)C(S)C(S)” (d, double-stranded; s, single-stranded) as a consensus sequence for RAG-induced breaks at single-/double-strand DNA transitions. Such a consensus sequence motif is useful for explaining RAG cleavage on other types of DNA structures described in the literature. Therefore, the mechanism of RAG cleavage described here could explain facets of chromosomal rearrangements specific to lymphoid tissues leading to genomic instability.

On breaks of the Indian monsoon

For over a century, the term break has been used for spells in which the rainfall over the Indian monsoon zone is interrupted. The phenomenon of 'break monsoon' is of great interest because long intense breaks are often associated with poor monsoon seasons. Such breaks have distinct circulation characteristics (heat trough type circulation) and have a large impact on rainfed agriculture. Although interruption of the monsoon rainfall is considered to be the most important feature of the break monsoon, traditionally breaks have been identified on the basis of the surface pressure and wind patterns over the Indian region. We have defined breaks (and active spells) on the basis of rainfall over the monsoon zone. The rainfall criteria are chosen so as to ensure a large overlap with the traditional breaks documented by Ramamurthy (1969) and De et al (1998). We have identified these rainbreaks for 1901-89. We have also identified active spells on the basis of rainfall over the Indian monsoon zone. We have shown that the all-India summer monsoon rainfall is significantly negatively correlated with the number of rainbreak days (correlation coefficient -0.56) and significantly positively correlated with the number of active days (correlation coefficient 0.47). Thus the interannual variation of the all-India summer monsoon rainfall is shown to be related to the number of days of rainbreaks and active spells identified here. There have been several studies of breaks (and also active spells in several cases) identified on the basis of different criteria over regions differing in spatial scales (e.g., Webster et al 1998; Krishnan et al 2000; Goswami and Mohan 2000; and Annamalai and Slingo 2001). We find that there is considerable overlap between the rainbreaks we have identified and breaks based on the traditional definition. There is some overlap with the breaks identified by Krishnan et al (2000) but little overlap with breaks identified by Webster et al (1998). Further, there are three or four active-break cycles in a season according to Webster et al (1998) which implies a time scale of about 40 days for which Goswami and Mohan (2000), and Annamalai and Slingo'(2001) have studied breaks and active minus break fluctuations. On the other hand, neither the traditional breaks (Ramamurthy 1969; and De et al 1998) nor the rainbreaks occur every year. This suggests that the 'breaks' in these studies axe weak spells of the intraseasonal variation of the monsoon, which occur every year. We have derived the OLR and circulation patterns associated with rainbreaks and active spells and compared them with the patterns associated with breaks/active minus break spells from these studies. Inspite of differences in the patterns over the Indian region, there is one feature which is seen in the OLR anomaly patterns of breaks identified on the basis of different criteria as well as the rainbreaks identified in this paper viz., a quadrapole over the Asia-west Pacific region arising from anomalies opposite (same) in sign to those over the Indian region occurring over the equatorial Indian Ocean and northern tropical (equatorial) parts of the west Pacific. Thus it appears that this quadrapole is a basic feature of weak spells of the intraseasonal,variation over the Asia-west Pacific region. Since the rainbreaks are intense weak spells, this basic feature is also seen in the composite patterns of these breaks. We find that rainbreaks (active spells) are also associated with negative (positive) anomalies over a part of the cast Pacific suggesting that the convection over the Indian region is linked to that over the east Pacific not only on the interannual scale (as evinced by the link between the Indian summer monsoon rainfall and ENSO) but on the intraseasonal scale as well.

Processing of DNA double-stranded breaks and intermediates of recombination and repair by saccharomyces cerevisiae Mre11 and its stimulation by Rad50, Xrs2, and Sae2 proteins

Saccharomyces cerevisiae RAD50, MRE11, and XRS2 genes are essential for telomere length maintenance, cell cycle checkpoint signaling, meiotic recombination, and DNA double-stranded break (DSB) repair via nonhomologous end joining and homologous recombination. The DSB repair pathways that draw upon Mre11-Rad50-Xrs2 subunits are complex, so their mechanistic features remain poorly understood. Moreover, the molecular basis of DSB end resection in yeast mre11-nuclease deficient mutants and Mre11 nuclease-independent activation of ATM in mammals remains unknown and adds a new dimension to many unanswered questions about the mechanism of DSB repair. Here, we demonstrate that S. cerevisiae Mre11 (ScMre11) exhibits higher binding affinity for single-over double-stranded DNA and intermediates of recombination and repair and catalyzes robust unwinding of substrates possessing a 3' single-stranded DNA overhang but not of 5' overhangs or blunt-ended DNA fragments. Additional evidence disclosed that ScMre11 nuclease activity is dispensable for its DNA binding and unwinding activity, thus uncovering the molecular basis underlying DSB end processing in mre11 nuclease deficient mutants. Significantly, Rad50, Xrs2, and Sae2 potentiate the DNA unwinding activity of Mre11, thus underscoring functional interaction among the components of DSB end repair machinery. Our results also show that ScMre11 by itself binds to DSB ends, then promotes end bridging of duplex DNA, and directly interacts with Sae2. We discuss the implications of these results in the context of an alternative mechanism for DSB end processing and the generation of single-stranded DNA for DNA repair and homologous recombination.

Snaps and mends: DNA breaks and chromosomal translocations

Integrity in entirety is the preferred state of any organism. The temporal and spatial integrity of the genome ensures continued survival of a cell. DNA breakage is the first step towards creation of chromosomal translocations. In this review, we highlight the factors contributing towards the breakage of chromosomal DNA. It has been well-established that the structure and sequence of DNA play a critical role in selective fragility of the genome. Several non-B-DNA structures such as Z-DNA, cruciform DNA, G-quadruplexes, R loops and triplexes have been implicated in generation of genomic fragility leading to translocations. Similarly, specific sequences targeted by proteins such as Recombination Activating Genes and Activation Induced Cytidine Deaminase are involved in translocations. Processes that ensure the integrity of the genome through repair may lead to persistence of breakage and eventually translocations if their actions are anomalous. An insufficient supply of nucleotides and chromatin architecture may also play a critical role. This review focuses on a range of events with the potential to threaten the genomic integrity of a cell, leading to cancer.