Etablierung und Vergleich von Multi-Colour-Fish-Techniken und Multiplex Amplifiable Probe Hybridisation (MAPH) zur Detektion kryptischer Aberrationen der subtelomerischen Chromosomenregionen

Mikroskopisch nicht sichtbare und somit kryptische Chromosomenmutationen stellen eine bedeutende Ursache für mentale Retardierung (MR) und multiple Fehlbildungen beim Menschen dar. Als Regionen mit vermehrten kryptischen Aberrationen gelten die proximal an die Telomer-DNA angrenzenden subtelomerischen Chromosomenbereiche. Die hohe Rekombinationsfrequenz der Chromosomenenden sowie zahlreiche Sequenzhomologien zwischen nichthomologen Chromosomen, welche Fehlpaarungen begünstigen, lassen auf eine hohe Aberrationsrate dieser Regionen schließen. In Kombination mit der großen Gendichte der Subtelomere ist zu erwarten, dass Mutationen in diesen Bereichen bei einer Vielzahl genetisch bedingter Störungen bislang unklarer Genese ursächlich beteiligt sind. Die wichtige ätiologische Rolle der Subtelomeraberrationen wurde vor allem bei der idiopathischen MR, die 1-2% der Bevölkerung betrifft, aufgezeigt. Die Häufigkeit kryptischer Imbalancen wurde in dieser Patientengruppe mit durchschnittlich 4,6% beziffert. Die hohe Prävalenz dieser Störungen sowie die große Zahl von 43 zu untersuchenden euchromatischen Subtelomerregionen erfordern den Einsatz hocheffizienter Techniken. Ziel der vorliegenden Arbeit war die Entwicklung und der Vergleich verschiedener Methoden der Fluoreszenz in situ-Hybridisierung (FISH) und der Multiplex Amplifiable Probe Hybridisation (MAPH) zur Erfassung kryptischer Aberrationen der Subtelomerregionen. Die Untersuchungen erfolgten an einem Kollektiv von 66 Patienten mit MR. Zunächst wurde überprüft, ob die Untersuchungseffizienz der FISH durch eine Erhöhung der Anzahl gleichzeitig dargestellter Sonden gesteigert werden konnte. Dazu wurden zwei Multi-Colour-FISH-Techniken, die Sechs-Farben-FISH und die Subtelomer-COBRA-FISH (COBRA= Combined Binary Ratio Labelling) entwickelt, die eine simultane Analyse aller 43 euchromatischen Subtelomere in sieben bzw. zwei Hybridisierungen gewährleisten. Diese Methoden wurden sowohl untereinander als auch mit dem kommerziell erhältlichen „ToTelVysionTM Multi-Colour FISH Probe Panel“ verglichen, bei dem eine vollständige Subtelomeranalyse 15 Hybridisierungen erfordert. Die Erhöhung der Anzahl gleichzeitig dargestellter Sonden erbrachte aufgrund der zunehmenden methodischen Komplexität keine Steigerung der Untersuchungseffizienz. Eher erschien ein spezifischer Einsatz der FISH-Methoden in Abhängigkeit von der Menge und Qualität des vorhandenen Untersuchungsmaterials sinnvoll. Da sich die FISH aufgrund des hohen zeitlichen und methodischen Aufwandes nur bedingt für eine Analyse großer Kollektive eignet, wurde die Subtelomer-MAPH als Hochdurchsatzverfahren zur Subtelomeranalyse etabliert. Diese Methode ermöglicht die simultane Untersuchung sämtlicher Subtelomere bei zwölf Patienten gleichzeitig. Als molekulargenetisches Verfahren gewährleistet sie im Gegensatz zur FISH zwar keine exakte Bestimmung der chromosomalen Lokalisation der untersuchten Regionen, konnte jedoch in den eigenen Untersuchungen den Aufwand der FISH-Analysen, die sich nur noch auf Regionen mit auffälligen MAPH-Befunden beschränkten, um mehr als 98% verringern. Dies stellt einen wichtigen methodischen Fortschritt in der Diagnostik kryptischer Aberrationen der Subtelomere dar

Concordance of attachment representations in preschool siblings assessed by the attachment story completion task

Several studies have indicated only a modest concordance of attachment security in siblings in infancy. Until now, very little was known about the concordance of siblings’ attachment security beyond infancy, as assessed by the attachment story completion task. This cross-sectional study aims to examine the concordance of attachment representations of 38 first-born (4–7 years) and 38 second-born (3–5 years) siblings living in middle-class two-parent families. Personality factors and the level of parenting stress of the biological mothers (30–43 years) were analysed in relation to children’s attachment security. The results indicate a 43 % secure-insecure concordance rate between siblings’ attachment representations. Sibling’s gender correspondence, age differences and differences in parenting stress were not related to attachment concordance whereas gender of the first-born child was related to attachment concordance. The results also indicate that older children more frequently had secure attachment representations compared to younger children and that attachment insecurity was associated with greater negative impacts of life events, lower maternal life satisfaction and higher parenting stress. Our study indicates that siblings’ attachment representations may lack concordance even when siblings are assessed by the same method at the same time. If maternal and environmental factors are able to explain a substantial amount of variance in the attachment security of individual children, non-shared environmental factors might be underestimated when studying siblings’ attachment representations. The significant effect of age on children’s attachment representations found in this study suggests the need for future research on the stability of attachment representations during the preschool years

Prion Protein in Milk

BACKGROUND: Prions are known to cause transmissible spongiform encephalopathies (TSE) after accumulation in the central nervous system. There is increasing evidence that prions are also present in body fluids and that prion infection by blood transmission is possible. The low concentration of the proteinaceous agent in body fluids and its long incubation time complicate epidemiologic analysis and estimation of spreading and thus the risk of human infection. This situation is particularly unsatisfactory for food and pharmaceutical industries, given the lack of sensitive tools for monitoring the infectious agent. METHODOLOGY/PRINCIPAL FINDINGS: We have developed an adsorption matrix, Alicon PrioTrap®, which binds with high affinity and specificity to prion proteins. Thus we were able to identify prion protein (PrP(C))–the precursor of prions (PrP(Sc))–in milk from humans, cows, sheep, and goats. The absolute amount of PrP(C) differs between the species (from µg/l range in sheep to ng/l range in human milk). PrP(C) is also found in homogenised and pasteurised off-the-shelf milk, and even ultrahigh temperature treatment only partially diminishes endogenous PrP(C) concentration. CONCLUSIONS/SIGNIFICANCE: In view of a recent study showing evidence of prion replication occurring in the mammary gland of scrapie infected sheep suffering from mastitis, the appearance of PrP(C) in milk implies the possibility that milk of TSE-infected animals serves as source for PrP(Sc)

Evaluation of modelled climatologies of O₃, CO, water vapour and NOy_{y} in the upper troposphere–lower stratosphere using regular in situ observations by passenger aircraft

Evaluating global chemistry models in the upper troposphere–lower stratosphere (UTLS) is an important step toward an improved understanding of the chemical composition in this region. This composition is regularly sampled through in situ measurements based on passenger aircraft, in the framework of the In-service Aircraft for a Global Observing System (IAGOS) research infrastructure. This study focuses on the comparison of the IAGOS measurements in ozone, carbon monoxide (CO), nitrogen reactive species (NOy) and water vapour, with a 25-year simulation output from the LMDZ-OR-INCA chemistry–climate model. For this purpose, we present and apply an extension of the Interpol-IAGOS software that projects the IAGOS data onto any model grid, in order to derive a gridded IAGOS product and a masked (sub-sampled) model product that are directly comparable to one another. Climatologies are calculated in the upper troposphere (UT) and in the lower stratosphere (LS) separately but also in the UTLS as a whole, as a demonstration for the models that do not sort out the physical variables necessary to distinguish between the UT and the LS. In the northern extratropics, the comparison in the UTLS layer suggests that the geographical distribution in the tropopause height is well reproduced by the model. In the separated layers, the model simulates well the water vapour climatologies in the UT and the ozone climatologies in the LS. There are opposite biases in CO in both UT and LS, which suggests that the cross-tropopause transport is overestimated. The NOy observations highlight the difficulty of the model in parameterizing the lightning emissions. In the tropics, the upper-tropospheric climatologies are remarkably well simulated for water vapour. They also show realistic CO peaks due to biomass burning in the most convective systems, and the ozone latitudinal variations are well correlated between the observations and the model. Ozone is more sensitive to lightning emissions than to biomass burning emissions, whereas the CO sensitivity to biomass burning emissions strongly depends on location and season. The present study demonstrates that the Interpol-IAGOS software is a tool facilitating the assessment of global model simulations in the UTLS, which is potentially useful for any modelling experiment involving chemistry climate or chemistry transport models

Supraparticles for Bare‐Eye H2 Indication and Monitoring: Design, Working Principle, and Molecular Mobility

Abstract Indicators for H2 are crucial to ensure safety standards in a green hydrogen economy. Herein, the authors report micron‐scaled indicator supraparticles for real‐time monitoring and irreversible recording of H2 gas via a rapid eye‐readable two‐step color change. They are produced via spray‐drying SiO2 nanoparticles, AuPd nanoparticles, and indicator‐dye resazurin. The resulting gas‐accessible mesoporous supraparticle framework absorbs water from humid atmospheres to create a three‐phase‐system. In the presence of H2, the color of the supraparticle switches first irreversibly from purple to pink and further reversibly to a colorless state. In situ infrared spectroscopy measurements indicate that this color change originates from the (ir)reversible H2‐induced reduction of resazurin to resorufin and hydroresorufin. Further infrared spectroscopic measurements and molecular dynamics simulations elucidate that key to achieve this functionality is an established three‐phase‐system within the supraparticles, granting molecular mobility of resazurin. Water acts as transport medium to carry resazurin molecules towards the catalytically active AuPd nanoparticles. The advantages of the supraparticles are their small dimensions, affordable and scalable production, fast response times, straightforward bare‐eye detection, and the possibility of simultaneously monitoring H2 exposure in real‐time and ex post. Therefore, H2 indicator supraparticles are an attractive safety additive for leakage detection and localization in a H2 economy

The SIB Swiss Institute of Bioinformatics' resources: focus on curated databases

The SIB Swiss Institute of Bioinformatics (www.isb-sib.ch) provides world-class bioinformatics databases, software tools, services and training to the international life science community in academia and industry. These solutions allow life scientists to turn the exponentially growing amount of data into knowledge. Here, we provide an overview of SIB's resources and competence areas, with a strong focus on curated databases and SIB's most popular and widely used resources. In particular, SIB's Bioinformatics resource portal ExPASy features over 150 resources, including UniProtKB/Swiss-Prot, ENZYME, PROSITE, neXtProt, STRING, UniCarbKB, SugarBindDB, SwissRegulon, EPD, arrayMap, Bgee, SWISS-MODEL Repository, OMA, OrthoDB and other databases, which are briefly described in this article

Concordance of Attachment Representations in Preschool Siblings Assessed by the Attachment Story Completion Task

Several studies have indicated only a modest concordance of attachment security in siblings in infancy. Until now, very little was known about the concordance of siblings' attachment security beyond infancy, as assessed by the attachment story completion task. This cross-sectional study aims to examine the concordance of attachment representations of 38 first-born (4-7years) and 38 second-born (3-5years) siblings living in middle-class two-parent families. Personality factors and the level of parenting stress of the biological mothers (30-43years) were analysed in relation to children's attachment security. The results indicate a 43% secure-insecure concordance rate between siblings' attachment representations. Sibling's gender correspondence, age differences and differences in parenting stress were not related to attachment concordance whereas gender of the first-born child was related to attachment concordance. The results also indicate that older children more frequently had secure attachment representations compared to younger children and that attachment insecurity was associated with greater negative impacts of life events, lower maternal life satisfaction and higher parenting stress. Our study indicates that siblings' attachment representations may lack concordance even when siblings are assessed by the same method at the same time. If maternal and environmental factors are able to explain a substantial amount of variance in the attachment security of individual children, non-shared environmental factors might be underestimated when studying siblings' attachment representations. The significant effect of age on children's attachment representations found in this study suggests the need for future research on the stability of attachment representations during the preschool years

Problems of direct composite posterior restorations : a clinical study

The aim of this clinical study was to describe the quality of posterior composite restorations (n = 329) performed on a group of patients (n=219) during an observation period of three years at various intervals (6, 12, 18, 24 and 36 months) after application. The parameters were assessed both In vivo and In vitro using clinical examinations, impressions and photography according to modified FDI criteria. For the statistical analysis of the results, the Wilcoxon test with a significance level of p = 0.05 was applied. After three years, In vivo five from the seven parameters exhibited significant changes. Only "retention" and "approximal contact" remained unchanged. In vitro studied parameters "anatomical form", "occlusal contour/wear" and "approximal contact" did not result in any significant changes, however "marginal adaptation", "surface luster" and "overhangs" deteriorated significantly. In summary, the results of this study show that composite posterior restorations were clinically acceptable in terms of specific parameters. However, unsatisfactory results have arisen in relation to the handling of composites, stated In vivo and In vitro especially in the reconstruction of the marginal adaptation, surface and overfilling