The effect of in vitro heating on the distribution of nuclear matrix polypeptides in HeLa cells.

The in situ nuclear matrix was obtained from HeLa cells. After permeabilization with nonionic detergent, the resulting structures were incubated for 1 h at 37 degrees C to determine whether or not such an incubation might result in the redistribution of nuclear polypeptides which resisted extraction with buffers of high-ionic strength (1.6 M NaCl or 0.25 M (NH4)2SO4 as well as DNase I digestion. Using indirect immunofluorescence experiments and monoclonal antibodies we show that heating to 37 degrees C changes the distribution of a 160 kDa protein previously shown to be a component of the inner matrix network. On the other hand, a 125 kDa polypeptide was not affected at all by the incubation. Our results clearly indicate that the inclusion of a 37 degrees C incubation (for example during digestion with DNase I) in the protocol to obtain the in situ nuclear matrix can result in the formation of in vitro artifacts.

Carbon isotopic ratio analysis by gas chromatography/combustion/isotope ratio mass spectrometry for the detection of gamma-hydroxybutyric acid (GHB) administration to humans.

Since GHB (gamma-hydroxybutyric acid) is naturally produced in the human body, clinical and forensic toxicologists must be able to discriminate between endogenous levels and a concentration resulting from exposure. To suggest an alternative to the use of interpretative concentration cut-offs, the detection of exogenous GHB in urine specimens was investigated by means of gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). GHB was isolated from urinary matrix by successive purification on Oasis MCX and Bond Elute SAX solid-phase extraction (SPE) cartridges prior to high-performance liquid chromatography (HPLC) fractioning using an Atlantis dC18 column eluted with a mixture of formic acid and methanol. Subsequent intramolecular esterification of GHB leading to the formation of gamma-butyrolactone (GBL) was carried out to avoid introduction of additional carbon atoms for carbon isotopic ratio analysis. A precision of 0.3 per thousand was determined using this IRMS method for samples at GHB concentrations of 10 mg/L. The (13)C/(12)C ratios of GHB in samples of subjects exposed to the drug ranged from -32.1 to -42.1 per thousand, whereas the results obtained for samples containing GHB of endogenous origin at concentration levels less than 10 mg/L were in the range -23.5 to -27.0 per thousand. Therefore, these preliminary results show that a possible discrimination between endogenous and exogenous GHB can be made using carbon isotopic ratio analyses.

Growing up in Ireland. Key findings: 13-year-olds. No. 2. Physical activity and obesity among 13-year-olds.

This Key Finding reports on data from the second wave of interviews with Growing Up in Ireland's Child Cohort. The 8,568 children and their families were first interviewed when the children were 9 years old, and then at age 13 years, when 7,400 were reinterviewed between August 2011 and February 2012.The findings show that boys and young people from more socially advantaged backgrounds were more likely to exercise, and that 13-year-olds who took more exercise (whether hard or light exercise) were less likely to be overweight or obese.While most of the young people in Growing Up in Ireland maintained a healthy weight over time, one in four was either overweight or obese, a finding similar to that at 9 years. Girls were also more likely to be classified as overweight or obese than boys. The majority of 13-year-olds were quite positive about their physical appearance, although a quarter rated themselves as below average in this respect, and girls tended to be less positive about their body image than boys. Dieting behaviours had also become evident at 13.To understand more fully the origins and course of overweight and obesity, the descriptive data in this Key Finding can be used in more complex analyses drawing on the rich data available on the child, family and other important contextual variables.This resource was contributed by the National Documentation Centre on Drug Use.

Garnet growth in the metasediments of the Zermatt-Saas Fee zone (western alps)

The Zermatt-Saas Fee Zone (ZSZ) in the Western Alps consists of multiple slices of ultramafic, mafic and metasedimentary rocks. They represent the remnants of the Mesozoic Piemonte-Ligurian oceanic basin which was subducted to eclogite facies conditions with peak pressures and temperatures of up to 20-28 kbar and 550-630 °C, followed by a greenschist overprint during exhumation. Previous studies, emphasizing on isotopie geochronology and modeling of REE-behavior in garnets from mafic eclogites, suggest that the ZSZ is buildup of tectonic slices which underwent a protracted diachronous subduction followed by a rapid synchronous exhumation. In this study Rb/Sr geochronology is applied to phengite included in garnets from metasediments of two different slices of the ZSZ to date garnet growth. Inclusion ages for 2 metapelitic samples from the same locality from the first slice are 44.25 ± 0.48 Ma and 43.19 ± 0.32 Ma. Those are about 4 Ma older than the corresponding matrix mica ages of respectively 40.02 ± 0.13 Ma and 39.55 ± 0.25 Ma. The inclusion age for a third calcschist sample, collected from a second slice, is 40.58 ± 0.24 Ma and the matrix age is 39.8 ± 1.5 Ma. The results show that garnet effectively functioned as a shield, preventing a reset of the Rb/Sr isotopie clock in the included phengites to temperatures well above the closure of Sr in mica. The results are consistent with the results of former studies on the ZSZ using both Lu/Hf and Sm/Nd geochronology on mafic eclogites. They confirm that at least parts of the ZSZ underwent close to peak metamorphic HP conditions younger than 43 m.y. ago before being rapidly exhumed about 40 m.y. ago. Fluid infiltration in rocks of the second slice occurred likely close to the peak metamorphic conditions, resulting in rapid growth of garnets. Similar calcschists from the same slice contain two distinct types of porphyroblast garnets with indications of multiple growth pulses and resorption indicated by truncated chemical zoning patterns. In-situ oxygen isotope Sensitive High Resolution Ion Microprobe (SHRIMP) analyses along profiles on central sections of the garnets reveal variations of up to 5 %o in individual garnets. The complex compositional zoning and graphite inclusion patterns as well as the variations in oxygen isotopes correspond to growing under changing fluid composition conditions caused by external infiltrated fluids. The ultramafic and mafic rocks, which were subducted along with the sediments and form the volumetrically most important part of the ZSZ, are the likely source of those mainly aqueous fluids. - La Zone de Zermatt-Saas Fee (ZZS) est constituée de multiples écailles de roches ultramafiques, mafiques et méta-sédimentaires. Cette zone, qui affleure dans les Alpes occidentales, représente les restes du basin océanique Piémontais-Ligurien d'âge mésozoïque. Lors de la subduction de ce basin océanique à l'Eocène, les différentes roches composant le planché océanique ont atteint les conditions du faciès éclogitique avec des pressions et des températures maximales estimées entre 20 - 28 kbar et 550 - 630 °C respectivement, avant de subir une rétrogression au faciès schiste vert pendant l'exhumation. Différentes études antérieures combinant la géochronologie isotopique et la modélisation des mécanismes gouvernant l'incorporation des terres rares dans les grenats des éclogites mafiques, suggèrent que la ZZS ne correspond pas à une seule unité, mais est constituée de différentes écailles tectoniques qui ont subi une subduction prolongée et diachrone suivie d'une exhumation rapide et synchrone. Afin de tester cette hypothèse, j'ai daté, dans cette étude, des phengites incluses dans les grenats des méta-sédiments de deux différentes écailles tectoniques de la ZZS, afin de dater la croissance relative de ces grenats. Pour cela j'ai utilisé la méthode géochronologique basée sur la décroissance du Rb87 en Sr87. J'ai daté trois échantillons de deux différentes écailles. Les premiers deux échantillons proviennent de Triftji, au nord du Breithorn, d'une première écaille dont les méta-sédiments sont caractérisés par des bandes méta-pélitiques à grenat et des calcschistes. Le troisième échantillon a été collectionné au Riffelberg, dans une écaille dont les méta-sédiments sont essentiellement des calcschistes qui sont mélangés avec des roches mafiques et des serpentinites. Ce mélange se trouve au-dessus de la grande masse de serpentinites qui forment le Riffelhorn, le Trockenersteg et le Breithorn, et qui est connu sous le nom de la Zone de mélange de Riffelberg (Bearth, 1953). Les inclusions dans les grenats de deux échantillons méta-pélitiques de la première écaille sont datées à 44.25 ± 0.48 Ma et à 43.19 ± 0.32 Ma. Ces âges sont à peu près 4 Ma plus vieux que les âges obtenus sur les phengites provenant de la matrice de ces mêmes échantillons qui donnent des âges de 40.02 ± 0.13 Ma et 39.55 ± 0.25 Ma respectivement. Les inclusions de phengite dans les grenats appartenant à un calcschiste de la deuxième écaille ont un âge de 40.58 ± 0.24 Ma alors que les phengites de la matrice ont un âge de 39.8 ± 1.5 Ma. Pour expliquer ces différences d'âge entre les phengites incluses dans le grenat et les phengites provenant de la matrice, nous suggérons que la cristallisation de grenat ait permis d'isoler ces phengites et de les préserver de tous rééquilibrage lors de la suite du chemin métamorphique prograde, puis rétrograde. Ceci est particulièrement important pour expliquer l'absence de rééquilibrage des phengites dans des conditions de températures supérieures à la température de fermeture du système Rb/Sr pour les phengites. Les phengites en inclusions n'ayant pas pu être datées individuellement, nous interprétons l'âge de 44 Ma pour les inclusions de phengite comme un âge moyen pour l'incorporation de ces phengites dans le grenat. Ces résultats sont cohérents avec les résultats des études antérieures de la ZZS utilisant les systèmes isotopiques de Sm/Nd et Lu/Hf sur des eclogites mafiques. ils confirment qu'aux moins une partie de la ZZS a subi des conditions de pression et de température maximale il y a moins de 44 à 42 Ma avant d'être rapidement exhumée à des conditions métamorphiques du faciès schiste vert supérieur autour de 40 Ma. Cette étude détaillée des grenats a permis, également, de mettre en évidence le rôle des fluides durant le métamorphisme prograde. En effet, si tous les grenats montrent des puises de croissance et de résorption, on peut distinguer, dans différents calcschists provenant de la deuxième écaille, deux types distincts de porphyroblast de grenat en fonction de la présence ou non d'inclusions de graphite. Nous lions ces puises de croissances/résorptions ainsi que la présence ou l'absence de graphite en inclusion dans les grenats à l'infiltration de fluides dans le système, et ceci durant tous le chemin prograde mais plus particulièrement proche et éventuellement peu après du pic du métamorphisme comme le suggère l'âge de 40 Ma mesuré dans les inclusions de phengites de l'échantillon du Riffelberg. Des analyses in-situ d'isotopes d'oxygène réalisé à l'aide de la SHRIMP (Sensitive High Resolution Ion Microprobe) dans des coupes centrales des grenats indiquent des variations jusqu'à 5 %o au sein même d'un grenat. Les motifs de zonations chimiques et d'inclusions de graphite complexes, ainsi que les variations du δ180 correspondent à une croissance de grenat sous des conditions de fluides changeantes dues aux infiltrations de fluides externes. Nous lions l'origine de ces fluides aqueux aux unités ultramafiques et mafiques qui ont été subductés avec les méta-sédiments ; unités ultramafiques et mafiques qui forment la partie volumétrique la plus importante de la ZZS.

Inflammatory effects of snake venom metalloproteinases

Metalloproteinases are abundant enzymes in crotaline and viperine snake venoms. They are relevant in the pathophysiology of envenomation, being responsible for local and systemic hemorrhage frequently observed in the victims. Snake venom metalloproteinases (SVMP) are zinc-dependent enzymes of varying molecular weights having multidomain organization. Some SVMP comprise only the proteinase domain, whereas others also contain a disintegrin-like domain, cysteine-rich, and lectin domains. They have strong structural similarities with both mammalian matrix metalloproteinases (MMP) and members of ADAMs (a disintegrin and metalloproteinase) group. Besides hemorrhage, snake venom metalloproteinase induce local myonecrosis, skin damage, and inflammatory reaction in experimental models. Local inflammation is an important characteristic of snakebite envenomations inflicted by viperine and crotaline snake species. Thus, in the recent years there is a growing effort to understand the mechanisms responsible for SVMP-induced inflammatory reaction and the structural determinants of this effect. This short review focuses the inflammatory effects evoked by SVMP.

High constant incidence rates of second primary cancers of the head and neck: a pooled analysis of 13 cancer registries.

Scanty data are available on the incidence (i.e., the absolute risk) of second cancers of the head and neck (HN) and its pattern with age. We investigated this issue using data from a multicentric study of 13 population-based cancer registries from Europe, Canada, Australia and Singapore for the years 1943-2000. A total of 99,257 patients had a first primary HN cancer (15,985 tongue, 22,378 mouth, 20,758 pharyngeal, and 40,190 laryngeal cancer), contributing to 489,855 person-years of follow-up. A total of 1,294 of the patients (1.3%) were diagnosed with second HN cancers (342 tongue, 345 mouth, 418 pharynx and 189 larynx). Male incidence rates of first HN cancer steeply increased from 0.68/100,000 at age 30-34 to 46.2/100,000 at age 70-74, and leveled off at older age; female incidence increased from 0.50/100,000 at age 30-34 to 16.5/100,000 at age 80-84. However, age-specific incidence of second HN cancers after a first HN cancer in men was around 200-300/100,000 between age 40-44 and age 70-74 and tended to decline at subsequent ages (150/100,000 at age 80-84); in women, incidence of second HN cancers was around 200-300/100,000 between age 45-49 and 80-84. The patterns of age-specific incidence were consistent for different subsites of second HN cancer and sexes; moreover, they were similar for age-specific incidence of first primary HN cancer in patients who subsequently developed a second HN cancer. The incidence of second HN cancers does not increase with age, but remains constant, or if anything, decreases with advancing age.

NI Registry of Self-harm Annual Report 2012/13

The purpose of the registry is to improve understanding about self-harm and related behaviours in Northern Ireland. The information gathered will be used to monitor trends and patterns over time and, perhaps most importantly, will help shape the development of services and support to meet need. The information will also help provide trusts and the Department of Health, Social Services and Public Safety with a more accurate understanding of the impact of self-harm on Emergency Departments.

Regional Health and Social Care Personal and Public Involvement Forum: Annual Update Report 2012/13

Personal and Public Involvement (PPI) is an essential component in the delivery of truly person centred services.�It is also a statutory duty.�The PHA has leadership responsibilities in respect of the implementation of PPI across HSC.�One of the ways in which the PHA discharges that leadership function, is through the Regional HSC PPI Forum.�This body brings together all HSC organisations, working alongside service users and carers, to bring a focus on involvement.�It promotes the sharing of best practice, identifies and tackles issues of common concern and providers a platform for the active participation of service users and carers.� Each year in response to a Priorities for Action (PFA) target, the PHA, working with HSC partners, service users and carers in the Forum, develop an Annual Report on PPI work taken forward through the Forum.The report for 2012/13 details progress in a number of important areas such as training, development of standards etc.

Beating cervical cancer: information for girls in Year 9, aged 12-13 (English and eight translations)

This leaflet is distributed to girls in Year 9 and explains about the HPV vaccine, which can help protect against cervical cancer.

Hepatitis B - Could I be at risk? (English and 13 translations)

�This leaflet provides information on�Hepatitis�B - how the virus�is spread,�the symptoms of the virus and treatment available.

Effect of sphingosine and phorbol-12-myristate-13-acetate on the growth and dimethylsulfoxide-induced differentiation in the insect trypanosomatid Herpetomonas samuelpessoai

We investigated the effect of two modulators of protein kinase C, sphingosine and phorbol-12-myristate-13-acetate (PMA), on the growth and dimethylsulfoxide (DMSO)-induced differentiation in Herpetomonas samuelpessoai. Sphingosine did not stimulate the transformation of undifferentiated-promastigotes in differentiated-paramastigotes. PMA alone or in association with DMSO increased the number of paramastigotes in comparison to control cells. DMSO inhibited the parasite growth (35%) and several unusual morphological features resembling aberrant cell division were observed. Sphingosine did not significantly reduce the growth in contrast to PMA. Collectively, our results demonstrated that the reduction of the proliferation translates in an increase of the differentiation rate in the insect trypanosomatid H. samuelpessoai.

Use of the chicken lysozyme 5' matrix attachment region to generate high producer CHO cell lines.

Scaffold or matrix attachment region (S/MAR) genetic elements have previously been proposed to insulate transgenes from repressive effects linked to their site of integration within the host cell genome. We have evaluated their use in various stable transfection settings to increase the production of recombinant proteins such as monoclonal antibodies from Chinese hamster ovary (CHO) cell lines. Using the green fluorescent protein coding sequence, we show that S/MAR elements mediate a dual effect on the population of transfected cells. First, S/MAR elements almost fully abolish the occurrence of cell clones that express little transgene that may result from transgene integration in an unfavorable chromosomal environment. Second, they increase the overall expression of the transgene over the whole range of expression levels, allowing the detection of cells with significantly higher levels of transgene expression. An optimal setting was identified as the addition of a S/MAR element both in cis (on the transgene expression vector) and in trans (co-transfected on a separate plasmid). When used to express immunoglobulins, the S/MAR element enabled cell clones with high and stable levels of expression to be isolated following the analysis of a few cell lines generated without transgene amplification procedures.

The effect of a smoking ban introduction to environmental tobacco smoke exposure in Swiss hospitality workers : Abstracts of the 23rd Annual Conference of the International Society of Environmental Epidemiology (ISEE), September 13-16, 2011, Barcelona, Spain

Background and Aims: To protect the population from environmental tobacco smoke (ETS) Switzerland introduced a nationwide rather heterogeneous smoking ban in May 2010. The exposure situation of non-smoking hospitality workers before and after implementation of the new law is being assessed in a prospective cohort study. Methods: Exposure to ETS was measured using a novel method developed by the Institute for Work and Health in Lausanne. It is a passive sampler called MoNIC (Monitor of NICotine). The nicotine of the ETS is fixed onto a filter and transformed into salt of not volatile nicotine. Subsequently the number of passively smoked cigarettes is calculated. Badges were placed at the workplace as well as distributed to the participants for personal measuring. Additionally a salivary sample was taken to determine nicotine concentration. Results: At baseline Spearman's correlation between workplace and personal badge was 0.47. The average cigarette equivalents per day at the workplace obtained by badge significantly dropped from 5.1 (95%- CI: 2.4 to 7.9) at baseline to 0.3 (0.2 to 0.4) at first follow-up (n=29) three months later (p<0.001). For personal badges the number of passively smoked cigarettes declined from 1.5 (2.7 to 0.4) per day to 0.5 (0.3 to 0.8) (n=16).Salivary nicotine concentration in a subset of 13 participants who had worked on the day prior to the examination was 2.63 ng/ml before and 1.53 ng/ml after the ban (p=0.04). Spearman's correlation of salivary nicotine was 0.56 with workplace badge and 0.79 with personal badge concentrations. Conclusions: Workplace measurements clearly reflect the smoking regulation in a venue. The MoNIC badge proves to be a sensitive measuring device to determine personal ETS exposure and it is a demonstrative measure for communication with lay audiences and study participants as the number of passively smoked cigarettes is an easily conceivable result.