Conserved leucine residue in the head region of morbillivirus fusion protein regulates the large conformational change during fusion activity

Paramyxovirus cell entry is controlled by the concerted action of two viral envelope glycoproteins, the fusion (F) and the receptor-binding (H) proteins, which together with a cell surface receptor mediate plasma membrane fusion activity. The paramyxovirus F protein belongs to class I viral fusion proteins which typically contain two heptad repeat regions (HR). Particular to paramyxovirus F proteins is a long intervening sequence (IS) located between both HR domains. To investigate the role of the IS domain in regulating fusogenicity, we mutated in the canine distemper virus (CDV) F protein IS domain a highly conserved leucine residue (L372) previously reported to cause a hyperfusogenic phenotype. Beside one F mutant, which elicited significant defects in processing, transport competence, and fusogenicity, all remaining mutants were characterized by enhanced fusion activity despite normal or slightly impaired processing and cell surface targeting. Using anti-CDV-F monoclonal antibodies, modified conformational F states were detected in F mutants compared to the parental protein. Despite these structural differences, coimmunoprecipitation assays did not reveal any drastic modulation in F/H avidity of interaction. However, we found that F mutants had significantly enhanced fusogenicity at low temperature only, suggesting that they folded into conformations requiring less energy to activate fusion. Together, these data provide strong biochemical and functional evidence that the conserved leucine 372 at the base of the HRA coiled-coil of F(wt) controls the stabilization of the prefusogenic state, restraining the conformational switch and thereby preventing extensive cell-cell fusion activity.

Cultural Heritage Online? Settle It in the Country of Origin of the Work

This article examines the conditions under which a system of extended collective licensing (ECL) for the use of works contained in the collections of cultural heritage institutions (CHIs) participating in Europeana could function within a cross-border basis. ECL is understood as a form of collective rights management whereby the application of freely negotiated copyright licensing agreements between a user and a collective management organisation (“CMO”), is extended by law to non-members of the organisation. ECL regimes have already been put in place in a few Member States and so far, all have the ability to apply only on a national basis. This article proposes a mechanism that would allow works licensed under an ECL system in one territory of the European Union to be made available in all the territories of the Union. The proposal rests on the statutory recognition of the “country of origin” principle, as necessary and sufficient territory for the negotiation and application of an ECL solution for the rights clearance of works contained in the collection of a cultural heritage institution, including orphan works.

Phase diagram with an enhanced spin-glass region of the mixed Ising–XY magnet LiHo{x}Er{1-x}F{4}

We present the experimental phase diagram of LiHoxEr1-xF4, a dilution series of dipolar-coupled model magnets. The phase diagram was determined using a combination of ac susceptibility and neutron scattering. Three unique phases in addition to the Ising ferromagnet LiHoF4 and the XY antiferromagnet LiErF4 have been identified. Below x = 0.86, an embedded spin-glass phase is observed, where a spin glass exists within the ferromagnetic structure. Below x = 0.57, an Ising spin glass is observed consisting of frozen needlelike clusters. For x ∼ 0.3–0.1, an antiferromagnetically coupled spin glass occurs. A reduction of TC(x) for the ferromagnet is observed which disobeys the mean-field predictions that worked for LiHoxY1-xF4.

Structural rearrangements of the central region of the morbillivirus attachment protein stalk domain trigger F protein refolding for membrane fusion

It is unknown how receptor binding by the paramyxovirus attachment proteins (HN, H, or G) triggers the fusion (F) protein to fuse with the plasma membrane for cell entry. H-proteins of the morbillivirus genus consist of a stalk ectodomain supporting a cuboidal head; physiological oligomers consist of non-covalent dimer-of-dimers. We report here the successful engineering of intermolecular disulfide bonds within the central region (residues 91-115) of the morbillivirus H-stalk; a sub-domain that also encompasses the putative F-contacting section (residues 111-118). Remarkably, several intersubunit crosslinks abrogated membrane fusion, but bioactivity was restored under reducing conditions. This phenotype extended equally to H proteins derived from virulent and attenuated morbillivirus strains and was independent of the nature of the contacted receptor. Our data reveal that the morbillivirus H-stalk domain is composed of four tightly-packed subunits. Upon receptor binding, these subunits structurally rearrange, possibly inducing conformational changes within the central region of the stalk, which, in turn, promote fusion. Given that the fundamental architecture appears conserved among paramyxovirus attachment protein stalk domains, we predict that these motions may act as a universal paramyxovirus F-triggering mechanism.

Cotrimoxazole prophylaxis is associated with reduced risk of incident tuberculosis in participants in the Swiss HIV Cohort Study

Cotrimoxazole reduces mortality in HIV-infected adults with tuberculosis (TB), and in vitro data suggest potential anti-mycobacterial activity of cotrimoxazole. We aimed to evaluate whether prophylaxis with cotrimoxazole is associated with a decreased risk of incident TB in SHCS participants. We determined the incidence of TB per 1000 person-years from January 1992 to December 2012. Rates were analyzed separately in participants with current or no previous antiretroviral treatment (ART) using Poisson regression adjusted for CD4 cell count, sex, region of origin, injecting drug use, and age. 13,431 cohort participants contributed 107,549 person-years follow-up; 182 patients had incident TB; 132 (73%) before and 50 (27%) after ART initiation. The multivariable incidence rate ratios for cumulative cotrimoxazole exposure per year for persons with no previous and current ART were 0.70 (95% CI 0.55-0.89) and 0.87 (0.74-1.0) respectively. Cotrimoxazole may prevent the development of TB among HIV-positive persons, especially among those with no previous ART.

Somatic and cognitive domains of depression in an underserved region of Ecuador: some cultural considerations.

Not enough research efforts on depression have been carried out up to now in Latin America. The knowledge that has resulted from research activities in the United States or Europe offers limited generalizability to other regions of the world, including Latin America. In the Andean highlands of Ecuador, we found very high rates of moderate and severe depressive symptoms, a finding that must be interpreted within its cultural context. Somatic manifestations of depression predominated over cognitive manifestations, and higher education level was protective against depression. These findings call for an appreciation of culturally-specific manifestations of depression and the social factors that influence them. These factors must be further studied in order to give them the deserved priority, allocate resources appropriately, and formulate innovative psychosocial interventions.

Acculturation and cultural identity of immigrant youth: The impact of courses in the language and culture of origin

Career choices in the fields of science, technology, engineering, and mathematics (STEM) are favoured by men and often avoided by women; on the other hand, women tend to choose fields such as the social sciences. This not only leads to a shortage of employees with STEM degrees, but also reinforces the prejudice that certain (personality) characteristics are ‘typically female’ or ‘typically male’. Career orientation motives of young women and men can have important implications for gender (a-)typical career choices. However, there is little empirical research on the correlates of career orientation motives in young women in the field of STEM. This study seeks to address this gap by outlining the components of career orientation motives and showing relationships among them. Therefore, our results provide insight into the circumstances and conditions that are associated with academic and career choices.

Involvement of a region of 23S ribosomal RNA of {\it E. coli\/} in decoding of termination codons

Involvement of E. coli 23S ribosomal RNA (rRNA) in decoding of termination codons was first indicated by the characterization of a 23S rRNA mutant that causes UGA-specific nonsense suppression. The work described here was begun to test the hypothesis that more 23S rRNA suppressors of specific nonsense mutations can be isolated and that they would occur non-randomly in the rRNA genes and be clustered in specific, functionally significant regions of rRNA.^ Approximately 2 kilobases of the gene for 23S rRNA were subjected to PCR random mutagenesis and the amplified products screened for suppression of nonsense mutations in trpA. All of the suppressor mutations obtained were located in a thirty-nucleotide part of the GTPase center, a conserved rRNA sequence and structure, and they and others made in that region by site-directed mutagenesis were shown to be UGA-specific in their suppression of termination codon mutations. These results proved the initial hypothesis and demonstrated that a group of nucleotides in this region are involved in decoding of the UGA termination codon. Further, it was shown that limitation of cellular availability or synthesis of L11, a ribosomal protein that binds to the GTPase center rRNA, resulted in suppression of termination codon mutations, suggesting the direct involvement of L11 in termination in vivo.^ Finally, in vivo analysis of certain site-specific mutations made in the GTPase center RNA demonstrated that (a) the G$\cdot$A base pair closing the hexanucleotide hairpin loop was not essential for normal termination, (b) the "U-turn" structure in the 1093 to 1098 hexaloop is critical for normal termination, (c) nucleotides A1095 and A1067, necessary for the binding to ribosomes of thiostrepton, an antibiotic that inhibits polypeptide release factor binding to ribosomes in vitro, are also necessary for normal peptide chain termination in vivo, and (d) involvement of this region of rRNA in termination is determined by some unique subset structure that includes particular nucleotides rather than merely by a general structural feature of the GTPase center.^ This work advances the understanding of peptide chain termination by demonstrating that the GTPase region of 23S rRNA participates in recognition of termination codons, through an associated ribosomal protein and specific conserved nucleotides and structural motifs in its RNA. ^

What Limits Bacterial Production in the Suboxic Region of Permanently Ice-Covered Lake Bonney, Antarctica?

Bacterial production assays (thymidine incorporation rates) were used to evaluate the activity of heterotrophic bacteria at the chemocline region in both the East (ELB) and West (WLB) Lobes of permanently ice-covered Lake Bonney, in the Taylor Valley of Antarctica. The magnitude of activity varied dramatically within the depth interval of 1 to 2 m from moderate to very low levels below the chemocline, especially in the East Lobe, where chemical distributions indicate the absence of a normally functioning nitrogen cycle. Several parameters (e.g. addition of nutrients or chelators, dilution) were manipulated in incubation experiments in order to identify factors that would enhance activity in the suboxic deep waters of the East Lobe. Activity, in terms of thymidine incorporation, was consistently detected in the deep-water communities, implying that, although the water may be 'toxic', the cells remain viable. None of the treatments resulted in consistent enhancement of thymidine incorporation rates in samples from below the chemocline. Bacterial populations above the chemocline appear to be phosphorus-limited. The nature of the limitation, toxicity or inhibition that limits bacterial activity in the suboxic waters has not been identified.

When to see a doctor for common health problems: distribution patterns of functional health literacy across migrant populations in Switzerland

BACKGROUND Knowing when to seek professional help for health problems is considered an important aspect of health literacy. However, little is known about the distribution of help-seeking knowledge in the general population or specific subpopulations. METHODS We analysed data from the "Health Monitoring of the Swiss Migrant Population 2010" and used a short survey tool to study the distribution of help-seeking knowledge. We sampled members of four migrant groups (from Portugal, Turkey, Serbia and Kosovo; n = 2,614). Our tool contained 12 items that addressed common physical and psychological health problems. A total sum score measured help-seeking knowledge. Two sub-scores analysed knowledge related to potential overuse (minor symptoms) or potential underuse (major symptoms). We applied linear regression to show variations in help-seeking knowledge by age, sex, region of origin and length of stay. RESULTS Controlling for self-rated health, we found that region of origin, higher education, female gender and younger age were significantly associated with higher knowledge scores. CONCLUSIONS We present empirical evidence of unequal distribution of help-seeking knowledge across four migrant populations in Switzerland. Our findings contribute to current conceptual developments in health literacy, and provide starting points for future research.