QMPSF is sensitive and specific in the detection of NPHP1 heterozygous deletions

BACKGROUND: Nephronophthisis, an autosomal recessive nephropathy, is responsible for 10% of childhood chronic renal failure. The deletion of its major gene, NPHP1, with a minor allele frequency of 0.24% in the general population, is the most common mutation leading to a monogenic form of childhood chronic renal failure. It is challenging to detect it in the heterozygous state. We aimed to evaluate the sensitivity and the specificity of the quantitative multiplex PCR of short fluorescent fragments (QMPSF) in its detection. METHODS: After setting up the protocol of QMPSF, we validated it on 39 individuals diagnosed by multiplex ligation-dependent probe amplification (MLPA) with normal NPHP1 copy number (n=17), with heterozygous deletion (n=13, seven parents and six patients), or with homozygous deletion (n=9). To assess the rate of the deletions that arise from independent events, deleted alleles were haplotyped. RESULTS: The results of QMPSF and MLPA correlated perfectly in the identification of 76 heterozygously deleted and 56 homozygously deleted exons. The inter-experimental variability of the dosage quotient obtained by QMPSF was low: control, 1.05 (median; range, 0.86-1.33, n = 102 exons); heterozygous deletion, 0.51 (0.42-0.67, n = 76 exons); homozygous deletion, 0 (0-0, n = 56 exons). All patients harboring a heterozygous deletion were found to carry a hemizygous mutation. At least 15 out of 18 deletions appeared on different haplotypes and one deletion appeared de novo. CONCLUSIONS: The cost- and time-effective QMPSF has a 100% sensitivity and specificity in the detection of NPHP1 deletion. The potential de novo appearance of NPHP1 deletions makes its segregation analysis highly recommended in clinical practice

Social inequalities in changes in health-related behaviour among Slovak adolescents aged between 15 and 19: A longitudinal study

<p>Abstract</p> <p>Background</p> <p>Lower socioeconomic position is generally associated with higher rates of smoking and alcohol consumption and lower levels of physical activity. Health-related behaviour is usually established during late childhood and adolescence. The aim of this study is to explore changes in health-related behaviour in a cohort of adolescents aged between 15 and 19, overall and by socioeconomic position.</p> <p>Methods</p> <p>The sample consisted of 844 first-year students (42.8% males, baseline in 1998 – mean age 14.9, follow-up in 2002 – mean age 18.8) from 31 secondary schools located in Kosice, Slovakia. This study focuses on changes in adolescents' smoking, alcohol use, experience with marijuana and lack of physical exercise with regard to their socioeconomic position. Four indicators of socioeconomic position were used – adolescents' current education level and employment status, and the highest education level and highest occupational status of their parents. We first made cross tabulations of HRB with these four indicators, using McNemar's test to assess differences. Next, we used logistic regression to assess adjusted associations, using likelihood ratio tests to assess statistical significance.</p> <p>Results</p> <p>Statistically significant increases were found in all health-related behaviours. Among males, the most obvious socioeconomic gradient was found in smoking, both at age 15 and at 19. Variations in socioeconomic differences in health-related behaviour were more apparent among females. Although at age 15, almost no socioeconomic differences in health-related behaviour were found, at age 19 differences were found for almost all socioeconomic indicators. Among males, only traditional socioeconomic gradients were found (the lower the socioeconomic position, the higher the prevalence of potentially harmful health-related behaviour), while among females reverse socioeconomic gradients were also found.</p> <p>Conclusion</p> <p>We confirmed an increase in unhealthy health-related behaviour during adolescence. This increase was related to socioeconomic position, and was more apparent in females.</p

Defending the genome from the enemy within:mechanisms of retrotransposon suppression in the mouse germline

The viability of any species requires that the genome is kept stable as it is transmitted from generation to generation by the germ cells. One of the challenges to transgenerational genome stability is the potential mutagenic activity of transposable genetic elements, particularly retrotransposons. There are many different types of retrotransposon in mammalian genomes, and these target different points in germline development to amplify and integrate into new genomic locations. Germ cells, and their pluripotent developmental precursors, have evolved a variety of genome defence mechanisms that suppress retrotransposon activity and maintain genome stability across the generations. Here, we review recent advances in understanding how retrotransposon activity is suppressed in the mammalian germline, how genes involved in germline genome defence mechanisms are regulated, and the consequences of mutating these genome defence genes for the developing germline

Stochastic Drift in Mitochondrial DNA Point Mutations: A Novel Perspective Ex Silico

The mitochondrial free radical theory of aging (mFRTA) implicates Reactive Oxygen Species (ROS)-induced mutations of mitochondrial DNA (mtDNA) as a major cause of aging. However, fifty years after its inception, several of its premises are intensely debated. Much of this uncertainty is due to the large range of values in the reported experimental data, for example on oxidative damage and mutational burden in mtDNA. This is in part due to limitations with available measurement technologies. Here we show that sample preparations in some assays necessitating high dilution of DNA (single molecule level) may introduce significant statistical variability. Adding to this complexity is the intrinsically stochastic nature of cellular processes, which manifests in cells from the same tissue harboring varying mutation load. In conjunction, these random elements make the determination of the underlying mutation dynamics extremely challenging. Our in silico stochastic study reveals the effect of coupling the experimental variability and the intrinsic stochasticity of aging process in some of the reported experimental data. We also show that the stochastic nature of a de novo point mutation generated during embryonic development is a major contributor of different mutation burdens in the individuals of mouse population. Analysis of simulation results leads to several new insights on the relevance of mutation stochasticity in the context of dividing tissues and the plausibility of ROS ”vicious cycle” hypothesis

Degree of urbanization and gender differences in substance use among Slovak adolescents

Substance use among adolescents varies with gender and between countries. Urbanization may contribute to this. The aim of our study is to explore the association between the degree of urbanization and gender differences in adolescent smoking, binge drinking, and cannabis use. A cross-sectional questionnaire survey of Slovak adolescents was used (N = 3,493; mean age = 14.33), stratified by degree of urbanization. The effects of gender and urbanization of the area and their interaction on substance use (smoking, binge drinking, and cannabis) were analyzed using a logistic regression model adjusted for age. Gender and area and their interaction had statistically significant (p <0.01) associations with substance use. The lower the urbanization of the area, the less riskily females behaved. An exception was found in the case of binge drinking where the results of the interaction of gender and degree of urbanization were not significant for the second least urbanized area. Prevalence rate of substance use among girls increased along with an increasing degree of urbanization, while the prevalence rate of substance use among boys remained constant

Developing a tool to measure health worker motivation in district hospitals in Kenya

BACKGROUND: We wanted to try to account for worker motivation as a key factor that might affect the success of an intervention to improve implementation of health worker practices in eight district hospitals in Kenya. In the absence of available tools, we therefore aimed to develop a tool that could enable a rapid measurement of motivation at baseline and at subsequent points during the 18-month intervention study. METHODS: After a literature review, a self-administered questionnaire was developed to assess the outcomes and determinants of motivation of Kenyan government hospital staff. The initial questionnaire included 23 questions (from seven underlying constructs) related to motivational outcomes that were then used to construct a simpler tool to measure motivation. Parallel qualitative work was undertaken to assess the relevance of the questions chosen and the face validity of the tool. RESULTS: Six hundred eighty-four health workers completed the questionnaires at baseline. Reliability analysis and factor analysis were used to produce the simplified motivational index, which consisted of 10 equally-weighted items from three underlying factors. Scores on the 10-item index were closely correlated with scores for the 23-item index, indicating that in future rapid assessments might be based on the 10 questions alone. The 10-item motivation index was also able to identify statistically significant differences in mean health worker motivation scores between the study hospitals (p<0.001). The parallel qualitative work in general supported these conclusions and contributed to our understanding of the three identified components of motivation. CONCLUSION: The 10-item score developed may be useful to monitor changes in motivation over time within our study or be used for more extensive rapid assessments of health worker motivation in Kenya

Stres na radu i zdravlje medicinskih sestara u jedinicama intenzivne njege u Srbiji

The aim of this study was to identify and analyse professional stressors, evaluate the level of stress in nurses in Intensive Care Units (ICU), and assess the correlation between the perception of stress and psychological and somatic symptoms or diseases shown by nurses. The research, designed as a crosssectional study, was carried out in the Intensive Care Units (ICU), in health centres in Serbia. The sample population encompassed 1000 nurses. Expanded Nursing Stress Scale (ENSS) was used as the research instrument. ENSS revealed a valid metric characteristic within our sample population. Nurses from ICUs rated situations involving physical and psychological working environments as the most stressful ones, whereas situations related to social working environment were described as less stressful; however, the differences in the perception of stressfulness of these environments were minor. Socio-demographic determinants of the participants (age, marital status and education level) significantly affected the perception of stress at work. Significant differences in the perception of stressfulness of particular stress factors were observed among nurses with respect to psychological and somatic symptoms (such as headache, insomnia, fatigue, despair, lower back pain, mood swings etc.) and certain diseases (such as hypertension, myocardial infarction, stroke, diabetes mellitus etc). In view of permanent escalation of professional stressors, creating a supportive working environment is essential for positive health outcomes, prevention of job-related diseases and better protection of already ill nurses.Cilj je ovoga rada bio identifi cirati i analizirati profesionalne stresore, procijeniti razinu stresa kod medicinskih sestara u jedinicama intenzivne njege te procijeniti korelaciju između percepcije stresa i prisutnosti psiholoških i somatskih simptoma ili bolesti kod medicinskih sestara. Istraživanje je provedeno u obliku studije presjeka u Jedinicama intenzivne njege u zdravstvenim centrima u Srbiji. Uzorak se sastojao od 1000 medicinskih sestara-tehničara. Za procjenu i analizu profesionalnih stresora korišten je upitnik Expanded Nursing Stress Scale (ENSS), koji je pokazao validne metrijske karakteristike na našoj ispitanoj populaciji. Medicinske sestre u Jedinicama intenzivne njege ocijenile su situacije iz fizičkoga i psihološkoga radnog okruženja kao izrazito opterećujuće, a situacije iz socijalnoga radnog okruženja kao manje opterećujuće. Razlika u percepciji stresogenosti navedenih radnih okruženja nije bila statistički značajna. Sociodemografske determinante ispitanika (dob, bračno stanje i stupanj obrazovanja) značajno utječu na percepciju stresa na radnom mjestu. Utvrđena je statistički značajna razlika u opažanju stresogenosti pojedinih stresnih situacija na radnom mjestu između medicinskih sestara u odnosu na postojanje psihosomatskih simptoma (kao što su glavobolja, nesanica, umor, očaj, bol u leđima, česte promjene raspoloženja) ili određenih bolesti (kao što su povišena hipertenzija, infarkt miokarda, cerebrovaskularni inzult, šećerna bolest). Zbog sve izraženije prisutnosti profesionalnih stresora nužno je poduzeti određene strateške mjere kod medicinskih sestara u Jedinicama intenzivne njege. Strateške mjere podrazumijevaju unaprjeđenje psihosocijalne radne klime, što bi unaprijedilo njihovo zdravlje i spriječilo nastanak bolesti u svezi s radom, ali i omogućilo bolju zaštitu već oboljelim medicinskim sestrama