Non B-DNA structures and genomic instability associated with myotonic dystrophy type 2 (CCTG).(CAGG) repeats

There are many diseases associated with the expansion of DNA repeats in humans. Myotonic dystrophy type 2 is one of such diseases, characterized by expansions of a (CCTG)•(CAGG) repeat tract in intron 1 of zinc finger protein 9 (ZNF9) in chromosome 3q21.3. The DM2 repeat tract contains a flanking region 5' to the tract that consists of a polymorphic repetitive sequence (TG)14-25(TCTG)4-11(CCTG) n. The (CCTG)•(CAGG) repeat is typically 11-26 repeats in persons without the disease, but can expand up to 11,000 repeats in affected individuals, which is the largest expansion seen in DNA repeat diseases to date. This DNA tract remains one of the least characterized disease-associated DNA repeats, and mechanisms causing the repeat expansion in humans have yet to be elucidated. Alternative, non B-DNA structures formed by the expanded repeats are typical in DNA repeat expansion diseases. These sequences may promote instability of the repeat tracts. I determined that slipped strand structure formation occurs for (CCTG)•(CAGG) repeats at a length of 42 or more. In addition, Z-DNA structure forms in the flanking human sequence adjacent to the (CCTG)•(CAGG) repeat tract. I have also performed genetic assays in E. coli cells and results indicate that the (CCTG)•(CAGG) repeats are more similar to the highly unstable (CTG)•(CAG) repeat tracts seen in Huntington's disease and myotonic dystrophy type 1, than to those of the more stable (ATTCT)•(AGAAT) repeat tracts of spinocerebellar ataxia type 10. This instability, however, is RecA-independent in the (CCTG)•(CAGG) and (ATTCT)•(AGAAT) repeats, whereas the instability is RecA-dependent in the (CTG)•(CAG) repeats. Structural studies of the (CCTG)•(CAGG) repeat tract and the flanking sequence, as well as genetic selection assays may reveal the mechanisms responsible for the repeat instability in E. coli, and this may lead to a better understanding of the mechanisms contributing to the human disease state. ^

Genome -wide and locus -specific instability

The discovery of expanded simple repeated sequences causing or associated with human disease has lead to a new area of research involved in the elucidation of how the expanded repeat causes disease and how the repeat becomes unstable. ^ To study the genetic basis of the (CTG)n repeat instability in the DMPK gene in myotonic dystrophy (DM1) patients, somatic cell hybrids were constructed between the lymphocytes of DM1 patients and a variety of Chinese hamster ovary (CHO) cell DNA repair gene deficient mutants. By using small pool PCR (SP-PCR), the instability of the (CTG)n can be quantitated for both the frequency and sizes of length change mutations. ^ Additional SP-PCR analysis on 2/11 subclones generated from this original hybrid showed a marked increase in large repeat deletions, ∼50%. A bimodal distribution of repeats was seen around the progenitor allele and at a large deleted product (within the normal range) with no intermediate products present. ^ To determine if the repair capacity of the CHO cell led to a mutator phenotype in the hamster and hybrid clones, SP-PCR was also done on 3 hamster microsatellites in a variety of hamster cell backgrounds. No variant alleles were seen in over 2500 genome equivalents screened. ^ Human-hamster hybrids have long been shown to be chromosomally unstable, yet information about the stability of repeated sequences was not known. To test if repeat instability was associated with either intact or non-intact human chromosomes, more than 300 microsatellite repeats on 13 human chromosomes (intact and non-intact) were analyzed in eight hybrid cells. No variants were seen between the hybrid and patient alleles in the hybrids. ^ To identify whether DM1 patients have a previously undetected level of genome wide instability or if the instability is truly locus specific, SP-PCR was done on 6 human microsatellites within the patient used to make the hybrid cells. No variants were seen in over 1000 genomes screened. ^ These studies show that the somatic cell hybrid approach is a genetically stable system that allows for the determination of factors that could lead to changes in microsatellite instability. It also shows that there is something inherent about the DM1 expanded (CTG)n repeat that it is solely targeted by, as of yet, and unknown mechanism that causes the repeat to be unstable. (Abstract shortened by UMI.)^

Genome instability quantified in constitutive tissues as well as putatively stable tumor tissue in hereditary non -polyposis colon cancer patients by small pool PCR

Microsatellite instability (MSI) is a hallmark of the mutator phenotype associated with Hereditary Non-Polyposis Colon Cancer (HNPCC). The MSI-High (MSI-H) HNPCC population has been well characterized, but the microsatellite low and stable (MSI-L/MSS) HNPCC population is much less understood. We hypothesize there are significant levels of MSI in HNPCC DNA classified as MSI-L/MSS, but no single variant allele makes up a sufficient population in the tumor DNA to be detected by standard analysis. Finding variants would suggest there is a mutator phenotype for the MSI-L/MSS HNPCC population that is distinct from the MSI-H HNPCC populations. This study quantified and compared MSI in HNPCC patients previously shown to be MSI-H, MSI-L/MSS and an MSI-H older, sporadic colorectal cancer patient. Small-pool Polymerase Chain Reactions (SP-PCRs) were conducted where the DNAs from each sample and controls are diluted into multiple pools, each containing approximately single genome equivalents. At least 100 alleles/sample were studied at six microsatellite loci. Mutant fragments were identified, quantified, and compared using Poisson statistics. Most of the variants were small deletions or insertions, with more mutants being deletions, as has been previously described in yeast and transgenic mice. SP-PCR, where most of the pools contained only 3 or less fragments, enabled identification of variants too infrequent to be detected by large pool PCR. Mutant fragments in positive control MSI-H tumor samples ranged from 0.26 to 0.68 in at least 4 of the 6 loci tested and were consistent with their MSI-H status. In the so called MSS tumors and constitutive tissues (normal colon tissue, and PBLs) of all the HNPCC patients, low, but significant levels of MSI were seen in at least two of the loci studied. This phenomenon was not seen in the sporadic MSI constitutive tissues nor the normal controls and suggests haploinsufficiency, gain-of-function, or a dominant/negative basis of the instability in HNPCC patients carrying germline mutations for tumor suppressor genes. A different frequency and spectrum of mutant fragments suggests a different genetic basis (other than a major mutation in MLH1 or MSH2) for disease in MSI-L and MSS HNPCC patients. ^

Strike-slip faults mediate the rise of crustal-derived fluids and mud volcanism in the deep sea

We report on newly discovered mud volcanoes located at about 4500 m water depth 90 km west of the deformation front of the accretionary wedge of the Gulf of Cadiz, and thus outside of their typical geotectonic environment. Seismic data suggest that fluid flow is mediated by a >400-km-long strike-slip fault marking the transcurrent plate boundary between Africa and Eurasia. Geochemical data (Cl, B, Sr, 87Sr/86Sr, Delta18O, DeltaD) reveal that fluids originate in oceanic crust older than 140 Ma. On their rise to the surface, these fluids receive strong geochemical signals from recrystallization of Upper Jurassic carbonates and clay-mineral dehydration in younger terrigeneous units. At present, reports of mud volcanoes in similar deep-sea settings are rare, but given that the large area of transform-type plate boundaries has been barely investigated, such pathways of fluid discharge may provide an important, yet unappreciated link between the deeply buried oceanic crust and the deep ocean.

Work instability. Presentation of an empirical model about its psychological impact

Research on the impact that work instability has on workers has the limitation of assess the relations among different variables separately, without examining the possible mediation relationships that can exists between them. The aim of this article is to test a conceptual model of the mediating relations between the uneasiness due to work instability and the psychological impact, in the framework of interactive stress theory, conducting a Path Analysis. 191 workers participated on the study, with a mean age of 31 years-old (SD = 11). Results showed that the proposed model didn't fit to the data. Alternative models were explored, consistent with the original conceptual model and the empiric evidence. A new causal model is proposed, where Uneasiness due to Work Instability as an independent variable, Personal Strain and Personal Resources as intervenient variables, and Anger, Hopelessness, and Satisfaction as dependent ones. The theoretical and empirical importance of the resulting model is discussed.

Work instability perception in a sample of psychologists

Considering work instability as a contextual stressor we designed a specific instrument to assess how it is perceived by a group of psychologists: The Perceived Uneasiness in Work Instability - Psychologists Inventory (in Spanish, IMPIL-PS). The data were collected from a 44-subject sample, both male and female, residents of the City of Buenos Aires and Greater Buenos Aires. We present data referring to sample characteristics and the areas with the greatest impact of the stressor are indicated. Recent research on work instability as a contextual stressor point out its influence on subjects' performance and behaviour.

Work instability versus mental health

This contribution is part of a research on guidance and employment in La Plata , province of Buenos Aires (Argentina) undertaken jointly by the Chairs of Preventive Psychology and Vocational Guidance, both pertaining to the course of studies for Psychology at the National University of La Plata. This research is based on four axes, namely, education-work-social policies-health. This paper shall focus around the health axis, which is not provided with placement and employment services. Some unsystematical guidance experiences from the services of Adolescence and Mental Health are currently under way in the area. Research points to the existence of new demands of psychological treatment from a population ("the new poor") afraid of losing their jobs -or even unemployed- who was not in the habit of going to the public hospital. In the casuistry explored here, people afraid of losing their jobs present more psychosomatic complexities. Local population is also analysed and the said analysis is linked with several national and international research projects.

Measurements of velocity-dependent and slip-dependent frictional strength in laboratory friction experiments on natural samples from IODP Expedition 316 and ODP Hole 190-1174B

Slowslip forms part of the spectrum of fault behaviour between stable creep and destructive earthquakes. Slow slip occurs near the boundaries of large earthquake rupture zones and may sometimes trigger fast earthquakes. It is thought to occur in faults comprised of rocks that strengthen under fast slip rates, preventing rupture as a normal earthquake, or on faults that have elevated pore-fluid pressures. However, the processes that control slow rupture and the relationship between slow and normal earthquakes are enigmatic. Here we use laboratory experiments to simulate faulting in natural rock samples taken from shallow parts of the Nankai subduction zone, Japan, where very low-frequency earthquakes - a form of slow slip - have been observed.We find that the fault rocks exhibit decreasing strength over millimetre-scale slip distances rather than weakening due to increasing velocity. However, the sizes of the slip nucleation patches in our laboratory simulations are similar to those expected for the very lowfrequency earthquakes observed in Nankai. We therefore suggest that this type of fault-weakening behaviour may generate slow earthquakes. Owing to the similarity between the expected behaviour of slow earthquakes based on our data, and that of normal earthquakes during nucleation, we suggest that some types of slow slip may represent prematurely arrested earthquakes.

Work instability. Presentation of an empirical model about its psychological impact

Research on the impact that work instability has on workers has the limitation of assess the relations among different variables separately, without examining the possible mediation relationships that can exists between them. The aim of this article is to test a conceptual model of the mediating relations between the uneasiness due to work instability and the psychological impact, in the framework of interactive stress theory, conducting a Path Analysis. 191 workers participated on the study, with a mean age of 31 years-old (SD = 11). Results showed that the proposed model didn't fit to the data. Alternative models were explored, consistent with the original conceptual model and the empiric evidence. A new causal model is proposed, where Uneasiness due to Work Instability as an independent variable, Personal Strain and Personal Resources as intervenient variables, and Anger, Hopelessness, and Satisfaction as dependent ones. The theoretical and empirical importance of the resulting model is discussed.

Work instability perception in a sample of psychologists

Considering work instability as a contextual stressor we designed a specific instrument to assess how it is perceived by a group of psychologists: The Perceived Uneasiness in Work Instability - Psychologists Inventory (in Spanish, IMPIL-PS). The data were collected from a 44-subject sample, both male and female, residents of the City of Buenos Aires and Greater Buenos Aires. We present data referring to sample characteristics and the areas with the greatest impact of the stressor are indicated. Recent research on work instability as a contextual stressor point out its influence on subjects' performance and behaviour.

Work instability versus mental health

This contribution is part of a research on guidance and employment in La Plata , province of Buenos Aires (Argentina) undertaken jointly by the Chairs of Preventive Psychology and Vocational Guidance, both pertaining to the course of studies for Psychology at the National University of La Plata. This research is based on four axes, namely, education-work-social policies-health. This paper shall focus around the health axis, which is not provided with placement and employment services. Some unsystematical guidance experiences from the services of Adolescence and Mental Health are currently under way in the area. Research points to the existence of new demands of psychological treatment from a population ("the new poor") afraid of losing their jobs -or even unemployed- who was not in the habit of going to the public hospital. In the casuistry explored here, people afraid of losing their jobs present more psychosomatic complexities. Local population is also analysed and the said analysis is linked with several national and international research projects.

Work instability perception in a sample of psychologists

Considering work instability as a contextual stressor we designed a specific instrument to assess how it is perceived by a group of psychologists: The Perceived Uneasiness in Work Instability - Psychologists Inventory (in Spanish, IMPIL-PS). The data were collected from a 44-subject sample, both male and female, residents of the City of Buenos Aires and Greater Buenos Aires. We present data referring to sample characteristics and the areas with the greatest impact of the stressor are indicated. Recent research on work instability as a contextual stressor point out its influence on subjects' performance and behaviour.

Work instability. Presentation of an empirical model about its psychological impact

Research on the impact that work instability has on workers has the limitation of assess the relations among different variables separately, without examining the possible mediation relationships that can exists between them. The aim of this article is to test a conceptual model of the mediating relations between the uneasiness due to work instability and the psychological impact, in the framework of interactive stress theory, conducting a Path Analysis. 191 workers participated on the study, with a mean age of 31 years-old (SD = 11). Results showed that the proposed model didn't fit to the data. Alternative models were explored, consistent with the original conceptual model and the empiric evidence. A new causal model is proposed, where Uneasiness due to Work Instability as an independent variable, Personal Strain and Personal Resources as intervenient variables, and Anger, Hopelessness, and Satisfaction as dependent ones. The theoretical and empirical importance of the resulting model is discussed.

Work instability versus mental health

This contribution is part of a research on guidance and employment in La Plata , province of Buenos Aires (Argentina) undertaken jointly by the Chairs of Preventive Psychology and Vocational Guidance, both pertaining to the course of studies for Psychology at the National University of La Plata. This research is based on four axes, namely, education-work-social policies-health. This paper shall focus around the health axis, which is not provided with placement and employment services. Some unsystematical guidance experiences from the services of Adolescence and Mental Health are currently under way in the area. Research points to the existence of new demands of psychological treatment from a population ("the new poor") afraid of losing their jobs -or even unemployed- who was not in the habit of going to the public hospital. In the casuistry explored here, people afraid of losing their jobs present more psychosomatic complexities. Local population is also analysed and the said analysis is linked with several national and international research projects.