Selective distribution of lactate dehydrogenase isoenzymes in neurons and astrocytes of human brain.

In vertebrates, the interconversion of lactate and pyruvate is catalyzed by the enzyme lactate dehydrogenase. Two distinct subunits combine to form the five tetrameric isoenzymes of lactate dehydrogenase. The LDH-5 subunit (muscle type) has higher maximal velocity (Vmax) and is present in glycolytic tissues, favoring the formation of lactate from pyruvate. The LDH-1 subunit (heart type) is inhibited by pyruvate and therefore preferentially drives the reaction toward the production of pyruvate. There is mounting evidence indicating that during activation the brain resorts to the transient glycolytic processing of glucose. Indeed, transient lactate formation during physiological stimulation has been shown by 1H-magnetic resonance spectroscopy. However, since whole-brain arteriovenous studies under basal conditions indicate a virtually complete oxidation of glucose, the vast proportion of the lactate transiently formed during activation is likely to be oxidized. These in vivo data suggest that lactate may be formed in certain cells and oxidized in others. We therefore set out to determine whether the two isoforms of lactate dehydrogenase are localized to selective cell types in the human brain. We report here the production and characterization of two rat antisera, specific for the LDH-5 and LDH-1 subunits of lactate dehydrogenase, respectively. Immunohistochemical, immunodot, and western-blot analyses show that these antisera specifically recognize their homologous antigens. Immunohistochemistry on 10 control cases demonstrated a differential cellular distribution between both subunits in the hippocampus and occipital cortex: neurons are exclusively stained with the anti-LDH1 subunit while astrocytes are stained by both antibodies. These observations support the notion of a regulated lactate flux between astrocytes and neurons.

Selective inhibition of CTL activation by a dipalmitoyl-phospholipid that prevents the recruitment of signaling molecules to lipid rafts.

Antigen-specific T-cell activation implicates a redistribution of plasma membrane-bound molecules in lipid rafts, such as the coreceptors CD8 and CD4, the Src kinases Lek and Fyn, and the linker for activation of T cells (LAT), that results in the formation of signaling complexes. These molecules partition in lipid rafts because of palmitoylation of cytoplasmic, membrane proximal cysteines, which is essential for their functional integrity in T-cell activation. Here, we show that exogenous dipalmitoyl-phosphatidylethanolamine (DPPE), but not the related unsaturated dioleoyl-phosphatidylethanolamine (DOPE), partitions in lipid rafts. DPPE inhibits activation of CD8(+) T lymphocytes by sensitized syngeneic antigen-presenting cells or specific major histocompatibility complex (MHC) peptide tetramers, as indicated by esterase release and intracellular calcium mobilization. Cytotoxic, T lymphocyte (CTL)-target cell conjugate formation is not affected by DPPE, indicating that engagement of the T-cell receptor by its cognate ligand is intact in lipid-treated cells. In contrast to other agents known to block raft-dependent signaling, DPPE efficiently inhibits the MHC peptide-induced recruitment of palmitoylated signaling molecules to lipid rafts and CTL activation without affecting cell viability or lipid raft integrity.

Functional analysis of Ectodysplasin-A mutations causing selective tooth agenesis.

Mutations of the Ectodysplasin-A (EDA) gene are generally associated with the syndrome hypohidrotic ectodermal dysplasia (MIM 305100), but they can also manifest as selective, non-syndromic tooth agenesis (MIM300606). We have performed an in vitro functional analysis of six selective tooth agenesis-causing EDA mutations (one novel and five known) that are located in the C-terminal tumor necrosis factor homology domain of the protein. Our study reveals that expression, receptor binding or signaling capability of the mutant EDA1 proteins is only impaired in contrast to syndrome-causing mutations, which we have previously shown to abolish EDA1 expression, receptor binding or signaling. Our results support a model in which the development of the human dentition, especially of anterior teeth, requires the highest level of EDA-receptor signaling, whereas other ectodermal appendages, including posterior teeth, have less stringent requirements and form normally in response to EDA mutations with reduced activity.

Sex-biased dispersal in a migratory bat: a characterization using sex-specific demographic parameters.

We studied the noctule bat (Nyctalus noctula), in which the mitochondrial F(ST) is about 10 times that revealed by nuclear markers, to address two questions. We first verified whether random dispersal of one sex is compatible with highly contrasted mitochondrial and nuclear population structures. Using computer simulations, we then assessed the power of multilocus population differentiation tests when the expected population structure departs only slightly from panmixia. Using an island model with sex-specific demographic parameters, we found that random male dispersal is consistent with the population structure observed in the noctule. However, other parameter combinations are also compatible with the data. We computed the minimum sex bias in dispersal (at least 69% of the dispersing individuals are males), a result that would not be available if we had used more classical population genetic models. The power of multilocus population differentiation tests was unexpectedly high, the tests being significant in almost 100% of the replicates, although the observed population structure infered from nuclear markers was extremely low (F(ST) = 0.6%).

Suboptimal access to primary healthcare among street-based sex workers in southwest Switzerland.

OBJECTIVES: Street-based sex workers (SSWs) in Lausanne, Switzerland, are poorly characterised. We set out to quantify potential vulnerability factors in this population and to examine SSW healthcare use and unmet healthcare requirements. METHODS: We conducted a cross-sectional questionnaire-based survey among SSWs working in Lausanne's red light district between 1 February and 31 July 2010, examining SSW socio-demographic characteristics and factors related to their healthcare. RESULTS: We interviewed 50 SSWs (76% of those approached). A fifth conducted their interviews in French, the official language in Lausanne. 48 participants (96%) were migrants, of whom 33/48 (69%) held no residence permit. 22/50 (44%) had been educated beyond obligatory schooling. 28/50 (56%) had no health insurance. 18/50 (36%) had been victims of physical violence. While 36/50 (72%) had seen a doctor during the preceding 12 months, only 15/50 (30%) were aware of a free clinic for individuals without health insurance. Those unaware of free services consulted emergency departments or doctors outside Switzerland. Gynaecology, primary healthcare and dental services were most often listed as needed. Two individuals (of 50, 4%) disclosed positive HIV status; of the others, 24/48 (50%) had never had an HIV test. CONCLUSIONS: This vulnerable population comprises SSWs who, whether through mobility, insufficient education or language barriers, are unaware of services they are entitled to. With half the participants reporting no HIV testing, there is a need to enhance awareness of available facilities as well as to increase provision and uptake of HIV testing.

Abortion trends 1990-1999 in a Swiss region and determinants of abortion recurrence.

OBJECTIVE: To assess age- and nationality-specific trends in abortion rates over the last decade, and to describe women's characteristics, identifying risk factors for repeated abortion. METHODS: From 1990-1999, the Health Department of Canton Vaud (Switzerland) received 13'857 abortion requests from residents aged 14-49. Population data were obtained to compute rates. RESULTS: Both the number of abortions (1400 annually) as well as their rate (8.9 per thousand women [95% confidence interval (CI) 7.3-10.5]) were stable over the decade in question. The rate of abortion for foreign women, especially from ex-Yugoslavia and Africa, was twice that for Swiss women. Half of the requests came from single women, 43% had a low education level, and half were childless. The main reason for requesting termination of pregnancy was psychosocial (93%). The mean gestational age was 7.7 weeks (SD +/- 2.3), but 96% of requests were submitted before 12 weeks. Sixty-three percent of women reported that they had used no contraception, 36% the condom and 17% the pill. Among requests, the adjusted risk of repeated abortion (22% of abortion candidates) was greater among divorced/separated/widowed women (odds ratio [OR] 1.9 [95% CI 1.5-2.4]), unemployed women (OR 1.8 [95% CI 1.5-2.1]), and those who had not attended university (OR 1.6 [95% CI 1.1-2.2]). CONCLUSIONS: Although Swiss law only permitted abortion under strict conditions, this procedure was widely available in Vaud, which nevertheless has one of the lowest rates worldwide. Efforts must be intensified to ensure universal access to family planning services, especially for foreign women and adolescents. Professionals should also target "repeaters" to provide personalised counselling.

The selective migration of young graduates : which of them return to their rural home region and which do not?

This paper addresses the migration behaviours of young university graduates from a rural region in Switzerland. Based on a questionnaire survey, it compares graduates' current place of residence (i.e. whether or not they returned to their home region) with characteristics related to their socio-familial, migration and professional trajectories. The propensity to return varies not only according to labour market variables (employment opportunities), but also to other factors, some of which have even more influence than job opportunities. The graduates' life course position (kind of household), their partners' characteristics (level of education and home region) and their family background (socio-economic status and history of migration) all play a central role. On the whole, results show that migration appears as a selective and complex process embedded in the life course of graduates.

Single loci with major effects in fire ants : the effects of the complementary sex-determining locus and the green beard supergene on gene expression

Etant données la complexité et la redondance des réseaux de gènes influençant de nombreux phénotypes, l'étude des rares cas d'un locus unique ayant des effets importants sur de nombreux phénotypes peut fournir des informations cruciales sur l'évolution des traits complexes. Nous avons séquencé le génome de la fourmi de feu Solenopsis invicta pour étudier comment l'expression des gènes détermine les effets majeurs et étendus de deux loci uniques sur le phénotype. Le premier locus concerne la détermination du sexe par le modèle des allèles complémentaires. Ce locus est connu pour déterminer le sexe chez tous les hyménoptères mais n'a été caractérisé que chez les abeilles. Les hétérozygotes pour ce locus se développent en reines diploïdes (ou ouvrières stériles) alors que les homozygotes se développent en mâles diploïdes incapables de produire du sperme et les hémizygotes en mâles haploïdes fertiles. Nous avons comparé l'expression des gènes entre les reines et les deux types de mâles au stade pupe, ainsi que 1 et 11 jours après l'émergence. Nous avons trouvé un changement prononcé de l'expression des gènes chez les mâles diploïdes, passant de très proche de celle des reines au stade pupe à identique aux mâles haploïdes 11 jours après l'émergence. Cela signifie que les mâles diploïdes sont condamnés à être stériles parce que les effets après émergence du locus de détermination du sexe ne per¬mettent pas d'effacer les effets de la ploïdie sur l'expression des gènes pendant le stade pupe, quand la spermatogénèse prend place. Le second locus aux effets majeurs que nous avons étudié est le supergène dit "green beard", qui consiste en 616 gènes couvrant 55% d'un chromosome (13 Mb) et est caractérisé par une absence de recombinaison entre les deux variants du supergène : "Social B" et "Social b" (SB et Sb). Au travers de l'effet "green beard", par lequel les ouvrières avec le supergène Sb discriminent favorablement les reines qui partagent ce supergène de façon perceptible, le génotype des reines fondatrices au niveau de ce supergène détermine l'organisation de la colonie : soit elle contient une seule reine SB/SB, soit plusieurs reines SB/Sb. Nous avons montré que le chromosome Sb a évolué comme le chromosome Y, accumulant probablement des allèles favorables dans des colonies avec plusieurs reines mais défavorables dans des colonies avec une seule reine (cf. gènes sexuellement antagonistes), ainsi que des transposons et des séquences répéti¬tives. Nous avons également montré que le polymorphisme du supergène cause de grandes différences d'expression chez les ouvrières et particulièrement les reines mais pas chez les mâles. Pour comprendre comment le polymorphisme du supergène chez les reines peut affecter l'organisation de la colonie, nous avons comparé l'expression entre les génotypes SB/SB et SB/Sb chez des reines vierges (1 et 11 jours) et des reines matures. Nous avons montré que les reines SB/SB sur-régulent des gènes impliqués dans la reproduction, expli-quant pourquoi elle grandissent plus rapidement et peuvent fonder des colonies de façon indépendante, tandis que les reines SB/Sb (qui ne peuvent fonder une nouvelle colonie) sur-régulent des gènes de signalement chimique qui affectent l'organisation des colonies par l'effet "green beard". - Given the complexity and redundancy of the gene networks that underlie many pheno- types, the study of rare cases of a single locus having major effects on many phenotypes can give powerful insights into the evolution of complex traits. We sequenced the genome of Solenopsis invicta fire ants to study how gene expression mediates the widespread major effects of two single loci on phenotype. The first is the complementary sex-determining locus, which is known to exist in most Hymenoptera despite being characterized only for honeybees. Heterozygotes at this locus become diploid queens (or sterile workers), homozy¬gotes become aspermic diploid males, and hemizygotes become fertile haploid males. We compared gene expression between queens and both types of males in pupae and 1 and 11 days after eclosion. We found a pronounced shift in gene expression in diploid males, from being nearly identical to queens as pupae to identical to haploid males 11 days after eclosion. This means that diploid males are condemned to sterility because the overriding effects of the sex locus after eclosion cannot undo the ploidy effects on expression during the pupal stage, when spermatogenesis must be completed. The second locus with major ef¬fects that we studied was the so-called "green beard" supergene, which consists of 616 genes encompassing 55% of one chromosome (13 Mb), without recombination between the two variants "Social B" and "Social b" (SB and Sb) supergene. Through the green beard effect, i.e. workers with the Sb supergene discriminating in favor of queens who perceptibly share this supergene, the founding queen's genotype at the supergene determines colony organi¬zation: either headed by a single SB/SB queen or many SB/Sb queens. We show that the Sb chromosome evolved like a Y-chromosome, probably accumulating alleles beneficial in multi-queen colonies but disadvantageous in single-queen colonies (cf. sexually antagonistic genes), as well as transposons and repetitive sequences. We also show that the polymor¬phism of the supergene causes widespread expression differences in workers and especially queens but not in males. To understand how the polymorphism at the supergene in queen can transform colony organization, we compared the expression between SB/SB and SB/Sb virgin queens (1 and 11 days) and mother queens. We show that SB/SB queens up-regulate genes involved in reproduction, explaining why they mature faster and can found colonies independently, while SB/Sb queens (which cannot found colonies) up-regulate chemical signaling genes that can transform colonies through the green beard effect.

Selective expression of the V beta 14 T cell receptor on Leishmania guyanensis--specific CD8+ T cells during human infection.

Peripheral blood mononuclear cells from subjects never exposed to Leishmania were stimulated with Leishmania guyanensis. We demonstrated that L. guyanensis-stimulated CD8(+) T cells produced interferon (IFN)- gamma and preferentially expressed the V beta 14 T cell receptor (TCR) gene family. In addition, these cells expressed cutaneous lymphocyte antigen and CCR4 surface molecules, suggesting that they could migrate to the skin. Results obtained from the lesions of patients with localized cutaneous leishmaniaisis (LCL) showed that V beta 14 TCR expression was increased in most lesions (63.5%) and that expression of only a small number of V beta gene families (V beta 1, V beta 6, V beta 9, V beta 14, and V beta 24) was increased. The presence of V beta 14 T cells in tissue confirmed the migration of these cells to the lesion site. Thus, we propose the following sequence of events during infection with L. guyanensis. After initial exposure to L. guyanensis, CD8(+) T cells preferentially expressing the V beta 14 TCR and secreting IFN- gamma develop and circulate in the periphery. During the infection, these cells migrate to the skin at the site of the parasitic infection. The role of these V beta 14 CD8(+) T cells in resistance to infection remains to be determined conclusively.

Alpha 3 domain mutants of peptide/MHC class I multimers allow the selective isolation of high avidity tumor-reactive CD8 T cells.

The goal of adoptive T cell therapy in cancer is to provide effective antitumor immunity by transfer of selected populations of tumor Ag-specific T cells. Transfer of T cells with high TCR avidity is critical for in vivo efficacy. In this study, we demonstrate that fluorescent peptide/MHC class I multimeric complexes incorporating mutations in the alpha3 domain (D227K/T228A) that abrogate binding to the CD8 coreceptor can be used to selectively isolate tumor Ag-specific T cells of high functional avidity from both in vitro expanded and ex vivo T cell populations. Sorting, cloning, and expansion of alpha3 domain mutant multimer-positive CD8 T cells enabled rapid selection of high avidity tumor-reactive T cell clones. Our results are relevant for ex vivo identification and isolation of T cells with potent antitumor activity for adoptive T cell therapy.

Selective attention to emotional stimuli: What IQ and Openness do, and emotional intelligence does not

We examined how general intelligence, personality, and emotional intelligence-measured as an ability using the MSCEIT-predicted performance on a selective-attention task requiring participants to ignore distracting emotion information. We used a visual prime in which participants saw a pair of faces depicting emotions; their task was to focus on one of the faces (the target) while ignoring the other (the distractor). Next, participants categorized a string of letters (word or nonword), which was either congruent to the target or the distractor. The speed of response to categorizing the string was recorded. Given the emotional nature of the stimuli and the emotional information processing involved in the task, we were surprised to see that none of the MSCEIT branches predicted performance. However, general intelligence and openness to experience reduced response time.

Haben selektive COX-2-Hemmer unerwünschte renale und kardiovaskuläre Wirkungen [Do selective COX-2 inhibitors have adverse renal and cardiovascular effects?].

Selective cyclooxygenase-2-inhibitors (COX-2) were developed as an alternative to the non-steroidal anti-inflammatory drugs (NSAID) in order to reduce their known gastrointestinal and renal toxicity. Several recent studies have shown the complex mechanism of the cyclooxygenase-2. The inhibition of the COX-2 has effects on renal hemodynamics, renal salt and water retention and may increase the thromboembolic and therefore the cardiovascular risk. The renal toxicity of the COX-2 inhibitors is similar to that of traditional NSAID. Regarding these data, COX-2 inhibitors should be prescribed with much caution to high risk patients, that is, patients with renal failure and/or cardiovascular diseases.

Selective neurodegeneration induced in rotation-mediated aggregate cell cultures by a transient switch to stationary culture conditions: a potential model to study ischemia-related pathogenic mechanisms.

Aggregating brain cell cultures at an advanced maturational stage (20-21 days in vitro) were subjected for 1-3 h to anaerobic (hypoxic) and/or stationary (ischemic) conditions. After restoration of the normal culture conditions, cell loss was estimated by measuring the release of lactate dehydrogenase as well as the irreversible decrease of cell type-specific enzyme activities, total protein and DNA content. Ischemia for 2 h induced significant neuronal cell death. Hypoxia combined with ischemia affected both neuronal and glial cells to different degrees (GABAergic neurons>cholinergic neurons>astrocytes). Hypoxic and ischemic conditions greatly stimulated the uptake of 2-deoxy-D-glucose, indicating increased glucose consumption. Furthermore, glucose restriction (5.5 mM instead of 25 mM) dramatically increased the susceptibility of neuronal and glial cells to hypoxic and ischemic conditions. Glucose media concentrations below 2 mM caused selective neuronal cell death in otherwise normal culture conditions. GABAergic neurons showed a particularly high sensitivity to glucose restriction, hypoxia, and ischemia. The pattern of ischemia-induced changes in vitro showed many similarities to in vivo findings, suggesting that aggregating brain cell cultures provide a useful in vitro model to study pathogenic mechanisms related to brain ischemia.

Selective accumulation of differentiated FOXP3(+) CD4 (+) T cells in metastatic tumor lesions from melanoma patients compared to peripheral blood

Precise identification of regulatory T cells is crucial in the understanding of their role in human cancers. Here, we analyzed the frequency and phenotype of regulatory T cells (Tregs), in both healthy donors and melanoma patients, based on the expression of the transcription factor FOXP3, which, to date, is the most reliable marker for Tregs, at least in mice. We observed that FOXP3 expression is not confined to human CD25(+/high) CD4(+) T cells, and that these cells are not homogenously FOXP3(+). The circulating relative levels of FOXP3(+) CD4(+) T cells may fluctuate close to 2-fold over a short period of observation and are significantly higher in women than in men. Further, we showed that FOXP3(+) CD4(+) T cells are over-represented in peripheral blood of melanoma patients, as compared to healthy donors, and that they are even more enriched in tumor-infiltrated lymph nodes and at tumor sites, but not in normal lymph nodes. Interestingly, in melanoma patients, a significantly higher proportion of functional, antigen-experienced FOXP3(+) CD4(+) T was observed at tumor sites, compared to peripheral blood. Together, our data suggest that local accumulation and differentiation of Tregs is, at least in part, tumor-driven, and illustrate a reliable combination of markers for their monitoring in various clinical settings.