An integrated genetic and functional analysis of the role of type II transmembrane serine proteases (TMPRSSs) in hearing loss.

Building on our discovery that mutations in the transmembrane serine protease, TMPRSS3, cause nonsyndromic deafness, we have investigated the contribution of other TMPRSS family members to the auditory function. To identify which of the 16 known TMPRSS genes had a strong likelihood of involvement in hearing function, three types of biological evidence were examined: 1) expression in inner ear tissues; 2) location in a genomic interval that contains a yet unidentified gene for deafness; and 3) evaluation of hearing status of any available Tmprss knockout mouse strains. This analysis demonstrated that, besides TMPRSS3, another TMPRSS gene was essential for hearing and, indeed, mice deficient for Hepsin (Hpn) also known as Tmprss1 exhibited profound hearing loss. In addition, TMPRSS2, TMPRSS5, and CORIN, also named TMPRSS10, showed strong likelihood of involvement based on their inner ear expression and mapping position within deafness loci PKSR7, DFNB24, and DFNB25, respectively. These four TMPRSS genes were then screened for mutations in affected members of the DFNB24 and DFNB25 deafness families, and in a cohort of 362 sporadic deaf cases. This large mutation screen revealed numerous novel sequence variations including three potential pathogenic mutations in the TMPRSS5 gene. The mutant forms of TMPRSS5 showed reduced or absent proteolytic activity. Subsequently, TMPRSS genes with evidence of involvement in deafness were further characterized, and their sites of expression were determined. Tmprss1, 3, and 5 proteins were detected in spiral ganglion neurons. Tmprss3 was also present in the organ of Corti. TMPRSS1 and 3 proteins appeared stably anchored to the endoplasmic reticulum membranes, whereas TMPRSS5 was also detected at the plasma membrane. Collectively, these results provide evidence that TMPRSS1 and TMPRSS3 play and TMPRSS5 may play important and specific roles in hearing.

In vitro functional correction of Hermansky-Pudlak Syndrome type-1 by lentiviral-mediated gene transfer.

Hermansky-Pudlak syndrome (HPS) is a genetic disorder characterized by oculocutaneous albinism, bleeding tendency and susceptibility to pulmonary fibrosis. No curative therapy is available. Genetic correction directed to the lungs, bone marrow and/or gastro-intestinal tract might provide alternative forms of treatment for the diseases multi-systemic complications. We demonstrate that lentiviral-mediated gene transfer corrects the expression and function of the HPS1 gene in patient dermal melanocytes, which opens the way to development of gene therapy for HPS.

Correlates of frailty, prediction of functional decline and preventative approaches - selected results from the lausanne cohort 65+ (lc65+) study (Switzerland) (Lausanne) and the longitudinal urban cohort ageing study (Lucas) (Germany)

Overall introduction.- Longitudinal studies have been designed to investigate prospectively, from their beginning, the pathway leading from health to frailty and to disability. Knowledge about determinants of healthy ageing and health behaviour (resources) as well as risks of functional decline is required to propose appropriate preventative interventions. The functional status in older people is important considering clinical outcome in general, healthcare need and mortality. Part I.- Results and interventions from lucas (longitudinal urban cohort ageing study). Authors.- J. Anders, U. Dapp, L. Neumann, F. Pröfener, C. Minder, S. Golgert, A. Daubmann, K. Wegscheider,. W. von Renteln-Kruse Methods.- The LUCAS core project is a longitudinal cohort of urban community-dwelling people 60 years and older, recruited in 2000/2001. Further LUCAS projects are cross-sectional comparative and interventional studies (RCT). Results.- The emphasis will be on geriatric medical care in a population-based approach, discussing different forms of access, too. (Dapp et al. BMC Geriatrics 2012, 12:35; http://www.biomedcentral.com/1471-2318/12/35): - longitudinal data from the LUCAS urban cohort (n = 3.326) will be presented covering 10 years of observation, including the prediction of functional decline, need of nursing care, and mortality by using a self-filling screening tool; - interventions to prevent functional decline do focus on first (pre-clinical) signs of pre-frailty before entering the frailty-cascade ("Active Health Promotion in Old Age", "geriatric mobility centre") or disability ("home visits"). Conclusions.- The LUCAS research consortium was established to study particular aspects of functional competence, its changes with ageing, to detect pre-clinical signs of functional decline, and to address questions on how to maintain functional competence and to prevent adverse outcome in different settings. The multidimensional data base allows the exploration of several further questions. Gait performance was exmined by GAITRite®-System. Supported by the Federal Ministry for Education and Research (BMBF Funding No. 01ET1002A). Part II.- Selected results from the lausanne cohort 65+ (Lc65 + ) Study (Switzerland). Authors.- Prof Santos-Eggimann Brigitte, Dr Seematter-Bagnoud Laurence, Prof Büla Christophe, Dr Rochat Stéphane. Methods.- The Lc65+ cohort was launched in 2004 with the random selection of 3054 eligible individuals aged 65 to 70 (birth year 1934-1938) in the non-institutionalized population of Lausanne (Switzerland). Results.- Information is collected about life course social and health-related events, socio-economics, medical and psychosocial dimensions, lifestyle habits, limitations in activities of daily living, mobility impairments, and falls. Gait performance are objectively measured using body-fixed sensors. Frailty is assessed using Fried's frailty phenotype. Follow-up consists in annual self-completed questionnaires, as well as physical examination and physical and mental performance tests every three years. - Lausanne cohort 65+ (Lc65 + ): design and longitudinal outcomes. The baseline data collection was completed among 1422 participants in 2004-2005 through self-completed questionnaires, face-to-face interviews, physical examination and tests of mental and physical performances. Information about institutionalization, self-reported health services utilization, and death is also assessed. An additional random sample (n = 1525) of 65-70 years old subjects was recruited in 2009 (birth year 1939-1943). - lecture no 4: alcohol intake and gait parameters: prevalent and longitudinal association in the Lc65+ study. The association between alcohol intake and gait performance was investigated.

Alpha-band suppression in the visual word form area as a functional bottleneck to consciousness.

The current state of empirical investigations refers to consciousness as an all-or-none phenomenon. However, a recent theoretical account opens up this perspective by proposing a partial level (between nil and full) of conscious perception. In the well-studied case of single-word reading, short-lived exposure can trigger incomplete word-form recognition wherein letters fall short of forming a whole word in one's conscious perception thereby hindering word-meaning access and report. Hence, the processing from incomplete to complete word-form recognition straightforwardly mirrors a transition from partial to full-blown consciousness. We therefore hypothesized that this putative functional bottleneck to consciousness (i.e. the perceptual boundary between partial and full conscious perception) would emerge at a major key hub region for word-form recognition during reading, namely the left occipito-temporal junction. We applied a real-time staircase procedure and titrated subjective reports at the threshold between partial (letters) and full (whole word) conscious perception. This experimental approach allowed us to collect trials with identical physical stimulation, yet reflecting distinct perceptual experience levels. Oscillatory brain activity was monitored with magnetoencephalography and revealed that the transition from partial-to-full word-form perception was accompanied by alpha-band (7-11 Hz) power suppression in the posterior left occipito-temporal cortex. This modulation of rhythmic activity extended anteriorly towards the visual word form area (VWFA), a region whose selectivity for word-forms in perception is highly debated. The current findings provide electrophysiological evidence for a functional bottleneck to consciousness thereby empirically instantiating a recently proposed partial perspective on consciousness. Moreover, the findings provide an entirely new outlook on the functioning of the VWFA as a late bottleneck to full-blown conscious word-form perception.

Functional responses and heterogeneities: an experimental test with cladocerans

The functional response of predators is usually modelled as a function of absolute prey density. Arditi and Ginzburg have suggested that it should often depend instead on the prey available per capita of predators, i.e. on the prey/predator ratio. Theory suggests that these two forms of dependence are related to the degree of spatial and temporal heterogeneity. Experiments using four filter-feeding cladoceran species were designed to test this hypothesis and to investigate the relation between individual behaviour and population dynamics. The patterns of population abundance that the cladocerans reached at equilibrium match the expectation that species with homogeneous spatial behaviour follow prey-dependent dynamics while those with heterogeneous behaviour follow ratio-dependent dynamics.

Functional compensation of motor function in pre-symptomatic Huntington's disease.

Involuntary choreiform movements are a clinical hallmark of Huntington's disease. Studies in clinically affected patients suggest a shift of motor activations to parietal cortices in response to progressive neurodegeneration. Here, we studied pre-symptomatic gene carriers to examine the compensatory mechanisms that underlie the phenomenon of retained motor function in the presence of degenerative change. Fifteen pre-symptomatic gene carriers and 12 matched controls performed button presses paced by a metronome at either 0.5 or 2 Hz with four fingers of the right hand whilst being scanned with functional magnetic resonance imaging. Subjects pressed buttons either in the order of a previously learnt 10-item finger sequence, from left to right, or kept still. Error rates ranged from 2% to 7% in the pre-symptomatic gene carriers and from 0.5% to 4% in controls, depending on the condition. No significant difference in task performance was found between groups for any of the conditions. Activations in the supplementary motor area (SMA) and superior parietal lobe differed with gene status. Compared with healthy controls, gene carriers showed greater activations of left caudal SMA with all movement conditions. Activations correlated with increasing speed of movement were greater the closer the gene carriers were to estimated clinical diagnosis, defined by the onset of unequivocal motor signs. Activations associated with increased movement complexity (i.e. with the pre-learnt 10-item sequence) decreased in the rostral SMA with nearing diagnostic onset. The left superior parietal lobe showed reduced activation with increased movement complexity in gene carriers compared with controls, and in the right superior parietal lobe showed greater activations with all but the most demanding movements. We identified a complex pattern of motor compensation in pre-symptomatic gene carriers. The results show that preclinical compensation goes beyond a simple shift of activity from premotor to parietal regions involving multiple compensatory mechanisms in executive and cognitive motor areas. Critically, the pattern of motor compensation is flexible depending on the actual task demands on motor control.

Flexible colony-founding strategies in a socially polymorphic ant

In social insects the number of queens per nest varies greatly. One of the proximate causes of this variation may be that queens produced by multiple-queen colonies are generally smaller, and might thus be unable to found new colonies independently. We examined whether the social origin of queens and males influenced the colony-founding success of queens in the socially polymorphic ant Formica selysi. Queens originating from single-queen and multiple-queen colonies had similar survival rates and colony-founding success, be they alone or in two-queen associations. During the first 5 months, queens originating from single-queen colonies gave rise to more workers than queens originating from multiple-queen colonies. Pairs of queens were also more productive than single queens. However, these differences in productivity were transient, as all types of colonies had reached a similar size after 15 months. Mating between social forms was possible and did not decrease queen survival or colony productivity, compared to mating within social forms. Overall, these results indicate that queens from each social form are able to found colonies independently, at least under laboratory conditions. Moreover, gene flow between social forms is not restricted by mating or genetic incompatibilities. This flexibility in mating and colony founding helps to explain the maintenance of alternative social structures in sympatry and the absence of genetic differentiation between social forms.

Cloning, functional expression, and chromosomal localization of the human pancreatic islet glucose-dependent insulinotropic polypeptide receptor.

Glucose-dependent insulinotropic polypeptide (GIP) is a hormone secreted by the endocrine K-cells from the duodenum that stimulates glucose-induced insulin secretion. Here, we present the molecular characterization of the human pancreatic islet GIP receptor. cDNA clones for the GIP receptor were isolated from a human pancreatic islet cDNA library. They encoded two different forms of the receptor, which differed by a 27-amino acid insertion in the COOH-terminal cytoplasmic tail. The receptor protein sequence was 81% identical to that of the rat GIP receptor. When expressed in Chinese hamster lung fibroblasts, both forms of the receptor displayed high-affinity binding for GIP (180 and 600 pmol/l). GIP binding was displaced by < 20% by 1 mumol/l glucagon, glucagon-like peptide (GLP-I)(7-36) amide, vasoactive intestinal peptide, and secretin. However exendin-4 and exendin-(9-39) at 1 mumol/l displaced binding by approximately 70 and approximately 100% at 10 mumol/l. GIP binding to both forms of the receptor induced a dose-dependent increase in intracellular cAMP levels (EC50 values of 0.6-0.8 nmol/l) but no elevation of cytoplasmic calcium concentrations. Interestingly, both exendin-4 and exendin-(9-39) were antagonists of the receptor, inhibiting GIP-induced cAMP formation by up to 60% when present at a concentration of 10 mumol/l. Finally, the physical and genetic chromosomal localization of the receptor gene was determined to be on 19q13.3, close to the ApoC2 gene. These data will help study the physiology and pathophysiology of the human GIP receptor.

Global periodicity conditions for maps and recurrences via Normal Forms

We face the problem of characterizing the periodic cases in parametric families of (real or complex) rational diffeomorphisms having a fixed point. Our approach relies on the Normal Form Theory, to obtain necessary conditions for the existence of a formal linearization of the map, and on the introduction of a suitable rational parametrization of the parameters of the family. Using these tools we can find a finite set of values p for which the map can be p-periodic, reducing the problem of finding the parameters for which the periodic cases appear to simple computations. We apply our results to several two and three dimensional classes of polynomial or rational maps. In particular we find the global periodic cases for several Lyness type recurrences

On Short Exponential Sums Involving Fourier Coefficients of Holomorphic Cusp Forms

By an exponential sum of the Fourier coefficients of a holomorphic cusp form we mean the sum which is formed by first taking the Fourier series of the said form,then cutting the beginning and the tail away and considering the remaining sum on the real axis. For simplicity’s sake, typically the coefficients are normalized. However, this isn’t so important as the normalization can be done and removed simply by using partial summation. We improve the approximate functional equation for the exponential sums of the Fourier coefficients of the holomorphic cusp forms by giving an explicit upper bound for the error term appearing in the equation. The approximate functional equation is originally due to Jutila [9] and a crucial tool for transforming sums into shorter sums. This transformation changes the point of the real axis on which the sum is to be considered. We also improve known upper bounds for the size estimates of the exponential sums. For very short sums we do not obtain any better estimates than the very easy estimate obtained by multiplying the upper bound estimate for a Fourier coefficient (they are bounded by the divisor function as Deligne [2] showed) by the number of coefficients. This estimate is extremely rough as no possible cancellation is taken into account. However, with small sums, it is unclear whether there happens any remarkable amounts of cancellation.

Exploring the link between flexible working and organizational performance

The expansion of flexible work experienced since the 1980s in developed economies is consistent with a more generic trend towards organizational flexibility, which many authors see as essential in order to compete in the dynamic global environment (Volberda, 1998). From this point of view, the changing demands of the environment have forced organizations to seek the ability to adapt rapidly and effectively as a means to be successful or even to survive. In the quest for flexibility, every area of the organization has been scrutinized in order to render it as ¿agile¿ as possible. In the human resources arena, this analysis has led to the definition of diverse ¿flexible working practices¿ (FWP) that describe a wide range of employment practices, which differ from the traditional full-time job with a fixed salary and a permanent contract. These practices have been described using other terms, such as ¿alternative¿ (Polivka, 1996; Powell & Mainiero, 1999), ¿non-standard¿ (Kalleberg, 2000), or ¿atypical¿ (De Grip, Hoevenberg, &m Willems, 1997), which coincide in denoting their divergence from the most traditional forms of employment. This article will show that quite different practices have been embraced by the common term ¿flexible working practices.¿ Subsequently, the results of empirical research regarding the implications for organizational performance of a number of flexible practices will be commented on.

Structural, functional and immunological studies on a polymeric bacterial protein

The characterization of proteins from Brucella spp, the causative agent of brucellosis, has been the subject of intensive research. We have described an 18-kDa cytoplasmic protein of Brucella abortus and shown the potential usefulness of this protein as an antigen for the serologic diagnosis of brucellosis. The amino acid sequence of the protein showed a low but significant homology with that of lumazine synthases. Lumazine is an intermediate product in bacterial riboflavin biosynthesis. The recombinant form of the 18-kDa protein (expressed in E. coli) folds like the native Brucella protein and has lumazine-synthase enzymatic activity. Three-dimensional analysis by X-ray crystallography of the homolog Bacillus subtilis lumazine synthase has revealed that the enzyme forms an icosahedral capsid. Recombinant lumazine synthase from B. abortus was crystallized, diffracted X rays to 2.7-Å resolution at room temperature, and the structure successfully solved by molecular replacement procedures. The macromolecular assembly of the enzyme differs from that of the enzyme from B. subtilis. The Brucella enzyme remains pentameric (90 kDa) in its crystallographic form. Nonetheless, the active sites of the two enzymes are virtually identical at the structural level, indicating that inhibitors of these enzymes could be viable pharmaceuticals across a broad species range. We describe the structural reasons for the differences in their quaternary arrangement and also discuss the potential use of this protein as a target for the development of acellular vaccines.

An investigation into the functional role of the D1:1 and D1:2 polypeptides in photosystem II in cyanobacteria : the effect of changing PSI/PSII ratio on photoinhibition in Synechococcus sp. PCC7942 /

The cyanobacterium Synechococcus sp. PCC 7942 (Anacystis nidulans R2) adjusts its photosynthetic function by changing one of the polypeptides of photosystem II. This polypeptide, called Dl, is found in two forms in Synechococcus sp. PCC 7942. Changing the growth light conditions by increasing the light intensity to higher levels results in replacement of the original form of D 1 polypeptide, D 1: 1, with another form, D 1 :2. We investigated the role of these two polypeptides in two mutant strains, R2S2C3 (only Dl:l present) and R2Kl (only Dl:2 present) In cells with either high or low PSI/PSII. R2S2C3 cells had a lower amplitude for 77 K fluorescence emission at 695 nm than R2Kl cells. Picosecond fluorescence decay kinetics showed that R2S2C3 cells had shorter lifetimes than R2Kl cells. The lower yields and shorter lifetimes observed in the D 1 and Dl:2 containing cells. containing cells suggest that the presence of D 1: 1 results in more photochemical or non-photochemical quenching of excitation energy In PSII. One of the most likely mechanisms for the increased quenching in R2S2C3 cells could be an increased efficiency in the transfer of excitation energy from PSII to PSI. However, photophysical studies including 77 K fluorescence measurements and picosecond time resolved decay kinetics comparing low and high PSI/PSII cells did not support the hypothesis that D 1: 1 facilitates the dissipation of excess energy by energy transfer from PSII to PSI. In addition physiological studies of oxygen evolution measurements after photoinhibition treatments showed that the two mutant cells had no difference in their susceptibility to photoinhibition with either high PSI/PSII ratio or low PSI/PSII ratio. Again suggesting that, the energy transfer efficiency from PSII to PSI is likely not a factor in the differences between Dl:l and Dl:2 containing cells.

Winâ Foy : functional object-oriented programming language

This thesis will introduce a new strongly typed programming language utilizing Self types, named Win--*Foy, along with a suitable user interface designed specifically to highlight language features. The need for such a programming language is based on deficiencies found in programming languages that support both Self types and subtyping. Subtyping is a concept that is taken for granted by most software engineers programming in object-oriented languages. Subtyping supports subsumption but it does not support the inheritance of binary methods. Binary methods contain an argument of type Self, the same type as the object itself, in a contravariant position, i.e. as a parameter. There are several arguments in favour of introducing Self types into a programming language (11. This rationale led to the development of a relation that has become known as matching [4, 5). The matching relation does not support subsumption, however, it does support the inheritance of binary methods. Two forms of matching have been proposed (lJ. Specifically, these relations are known as higher-order matching and I-bound matching. Previous research on these relations indicates that the higher-order matching relation is both reflexive and transitive whereas the f-bound matching is reflexive but not transitive (7]. The higher-order matching relation provides significant flexibility regarding inheritance of methods that utilize or return values of the same type. This flexibility, in certain situations, can restrict the programmer from defining specific classes and methods which are based on constant values [21J. For this reason, the type This is used as a second reference to the type of the object that cannot, contrary to Self, be specialized in subclasses. F-bound matching allows a programmer to define a function that will work for all types of A', a subtype of an upper bound function of type A, with the result type being dependent on A'. The use of parametric polymorphism in f-bound matching provides a connection to subtyping in object-oriented languages. This thesis will contain two main sections. Firstly, significant details concerning deficiencies of the subtype relation and the need to introduce higher-order and f-bound matching relations into programming languages will be explored. Secondly, a new programming language named Win--*Foy Functional Object-Oriented Programming Language has been created, along with a suitable user interface, in order to facilitate experimentation by programmers regarding the matching relation. The construction of the programming language and the user interface will be explained in detail.

Neural substrates and functional connectivity associated with sleep-dependent and independent consolidation of new motor skills

La mémoire n’est pas un processus unitaire et est souvent divisée en deux catégories majeures: la mémoire déclarative (pour les faits) et procédurale (pour les habitudes et habiletés motrices). Pour perdurer, une trace mnésique doit passer par la consolidation, un processus par lequel elle devient plus robuste et moins susceptible à l’interférence. Le sommeil est connu comme jouant un rôle clé pour permettre le processus de consolidation, particulièrement pour la mémoire déclarative. Depuis plusieurs années cependant, son rôle est aussi reconnu pour la mémoire procédurale. Il est par contre intéressant de noter que ce ne sont pas tous les types de mémoire procédurale qui requiert le sommeil afin d’être consolidée. Entre autres, le sommeil semble nécessaire pour consolider un apprentissage de séquences motrices (s’apparentant à l’apprentissage du piano), mais pas un apprentissage d’adaptation visuomotrice (tel qu’apprendre à rouler à bicyclette). Parallèlement, l’apprentissage à long terme de ces deux types d’habiletés semble également sous-tendu par des circuits neuronaux distincts; c’est-à-dire un réseau cortico-striatal et cortico-cérébelleux respectivement. Toutefois, l’implication de ces réseaux dans le processus de consolidation comme tel demeure incertain. Le but de cette thèse est donc de mieux comprendre le rôle du sommeil, en contrôlant pour le simple passage du temps, dans la consolidation de ces deux types d’apprentissage, à l’aide de l’imagerie par résonnance magnétique fonctionnelle et d’analyses de connectivité cérébrale. Nos résultats comportementaux supportent l’idée que seul l’apprentissage séquentiel requiert le sommeil pour déclencher le processus de consolidation. Nous suggérons de plus que le putamen est fortement associé à ce processus. En revanche, les performances d’un apprentissage visuomoteur s’améliorent indépendamment du sommeil et sont de plus corrélées à une plus grande activation du cervelet. Finalement, en explorant l’effet du sommeil sur la connectivité cérébrale, nos résultats démontrent qu’en fait, un système cortico-striatal semble être plus intégré suite à la consolidation. C’est-à-dire que l’interaction au sein des régions du système est plus forte lorsque la consolidation a eu lieu, après une nuit de sommeil. En opposition, le simple passage du temps semble nuire à l’intégration de ce réseau cortico-striatal. En somme, nous avons pu élargir les connaissances quant au rôle du sommeil pour la mémoire procédurale, notamment en démontrant que ce ne sont pas tous les types d’apprentissages qui requièrent le sommeil pour amorcer le processus de consolidation. D’ailleurs, nous avons également démontré que cette dissociation de l’effet du sommeil est également reflétée par l’implication de deux réseaux cérébraux distincts. À savoir, un réseau cortico-striatal et un réseau cortico-cérébelleux pour la consolidation respective de l’apprentissage de séquence et d’adaptation visuomotrice. Enfin, nous suggérons que la consolidation durant le sommeil permet de protéger et favoriser une meilleure cohésion au sein du réseau cortico-striatal associé à notre tâche; un phénomène qui, s’il est retrouvé avec d’autres types d’apprentissage, pourrait être considéré comme un nouveau marqueur de la consolidation.