An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping of protein interactions and subcellular localizations

Protein-protein interactions govern almost all cellular functions. These complex networks of stable and transient associations can be mapped by affinity purification mass spectrometry (AP-MS) and complementary proximity-based labeling methods such as BioID. To exploit the advantages of both strategies, we here design and optimize an integrated approach combining AP-MS and BioID in a single construct, which we term MAC-tag. We systematically apply the MAC-tag approach to 18 subcellular and 3 sub-organelle localization markers, generating a molecular context database, which can be used to define a protein's molecular location. In addition, we show that combining the AP-MS and BioID results makes it possible to obtain interaction distances within a protein complex. Taken together, our integrated strategy enables the comprehensive mapping of the physical and functional interactions of proteins, defining their molecular context and improving our understanding of the cellular interactome.Peer reviewe

Atypical hemolytic uremic syndrome in the era of terminal complement inhibition: an observational cohort study

Historically, the majority of patients with complement-mediated atypical hemolytic uremic syndrome (CaHUS) progress to end-stage kidney disease (ESKD). Single-arm trials of eculizumab with a short follow-up suggested efficacy. We prove, for the first time to our knowledge, in a genotype matched CaHUS cohort that the 5-year cumulative estimate of ESKD-free survival improved from 39.5% in a control cohort to 85.5% in the eculizumab-treated cohort (hazard ratio, 4.95; 95% confidence interval [CI], 2.75-8.90; P = .000; number needed to treat, 2.17 [95% CI, 1.81-2.73]). The outcome of eculizumab treatment is associated with the underlying genotype. Lower serum creatinine, lower platelet count, lower blood pressure, and younger age at presentation as well as shorter time between presentation and the first dose of eculizumab were associated with estimated glomerular filtration rate >60 ml/min at 6 months in multivariate analysis. The rate of meningococcal infection in the treated cohort was 550 times greater than the background rate in the general population. The relapse rate upon eculizumab withdrawal was 1 per 9.5 person years for patients with a pathogenic mutation and 1 per 10.8 person years for those with a variant of uncertain significance. No relapses were recorded in 67.3 person years off eculizumab in those with no rare genetic variants. Eculizumab was restarted in 6 individuals with functioning kidneys in whom it had been stopped, with no individual progressing to ESKD. We demonstrated that biallelic pathogenic mutations in RNA-processing genes, including EXOSC3, encoding an essential part of the RNA exosome, cause eculizumab nonresponsive aHUS. Recessive HSD11B2 mutations causing apparent mineralocorticoid excess may also present with thrombotic microangiopathy

Effectiveness of interventions to improve the health and housing status of homeless people: a rapid systematic review

Background: Research on interventions to positively impact health and housing status of people who are homeless has received substantially increased attention over the past 5 years. This rapid review examines recent evidence regarding interventions that have been shown to improve the health of homeless people, with particular focus on the effect of these interventions on housing status. Methods: A total of 1,546 articles were identified by a structured search of five electronic databases, a hand search of grey literature and relevant journals, and contact with experts. Two reviewers independently screened the first 10% of titles and abstracts for relevance. Inter-rater reliability was high and as a result only one reviewer screened the remaining titles and abstracts. Articles were included if they were published between January 2004 and December 2009 and examined the effectiveness of an intervention to improve the health or healthcare utilization of people who were homeless, marginally housed, or at risk of homelessness. Two reviewers independently scored all relevant articles for quality. Results: Eighty-four relevant studies were identified; none were of strong quality while ten were rated of moderate quality. For homeless people with mental illness, provision of housing upon hospital discharge was effective in improving sustained housing. For homeless people with substance abuse issues or concurrent disorders, provision of housing was associated with decreased substance use, relapses from periods of substance abstinence, and health services utilization, and increased housing tenure. Abstinent dependent housing was more effective in supporting housing status, substance abstinence, and improved psychiatric outcomes than non-abstinence dependent housing or no housing. Provision of housing also improved health outcomes among homeless populations with HIV. Health promotion programs can decrease risk behaviours among homeless populations. Conclusions: These studies provide important new evidence regarding interventions to improve health, housing status, and access to healthcare for homeless populations. The additional studies included in this current review provide further support for earlier evidence which found that coordinated treatment programs for homeless persons with concurrent mental illness and substance misuse issues usually result in better health and access to healthcare than usual care. This review also provides a synthesis of existing evidence regarding interventions that specifically support homeless populations with HIV.Partial funding for this paper was provided to the Effective Public Health Practice Project by the Region of Peel, Canada

Recall of Nadir CD4 Cell Count and Most Recent HIV Viral Load Among HIV-Infected, Socially Marginalized Adults

© 2015 Springer Science+Business Media New York (outside the USA) Lower nadir CD4 cell counts and higher HIV viral loads are associated with increased risks of adverse events in the progression of HIV disease. In cases where medical records are inaccessible or incomplete, little evidence is available regarding whether nadir CDR cell count or HIV viral load is reliably reported in any patient population. We compare survey data collected from 207 HIV-infected individuals detained in San Francisco jails to data collected from electronic medical records (EMR) kept by the jails and community health providers. The sensitivity of self-reported nadir CD4 cell count less than 200 was 82 % [95 % confidence interval (CI) 68, 88], and the sensitivity of reporting an undetectable most recent HIV viral load was 93 % (95 % CI 84, 97). This suggests that in a highly socially marginalized population, nadir CD4 cell count and most recent HIV viral load are recalled accurately when compared to EMR

Estudo da mastigação e da deglutição em crianças e adolescentes com Sequência de Möbius Study of chewing and swallowing in children and teenagers with Möbius syndrome

OBJETIVOS: Descrever as alterações morfofuncionais nas funções de mastigação e deglutição, reconhecer as consistências alimentares mais bem aceitas e avaliar a presença de manobras compensatórias durante a deglutição em crianças com Sequência de Möbius. MÉTODOS: A casuística consistiu de oito crianças, sendo três do sexo feminino e cinco do sexo masculino, na faixa etária de cinco a 15 anos, diagnosticadas com Sequência de Möbius. As crianças foram avaliadas em situação de alimentação, nas quais foram oferecidos alimentos de consistência líquida, pastosa e sólida. Os dados coletados foram registrados em protocolo de avaliação adaptado por Guedes, Shintani e Cabello (2003). RESULTADOS: Os resultados obtidos demonstraram significância entre a condição de mobilidade de língua e o tipo de mastigação. As variáveis velocidade de mastigação e contração do masseter, também se mostraram dependentes (p=0,018), uma vez que não houve alteração da velocidade da mastigação quando a contração do masseter estava presente. O pão de queijo mostrou-se como a melhor consistência sólida, pois produziu um bolo mais coeso e menos espalhado na cavidade oral. CONCLUSÕES: As alterações morfofuncionais presentes nos pacientes com Sequência de Möbius contribuíram para a alteração da dinâmica da mastigação e deglutição. Toda a amostra fez uso de manobras compensatórias e, mesmo com a ausência dos sinais clínicos sugestivos de penetração/aspiração, estes pacientes devem ser submetidos à terapia fonoaudiológica, a fim de adequar os órgãos fonoarticulatórios na tentativa de proporcionar uma refeição mais prazerosa.<br>PURPOSE: To describe morphofunctional alterations in the functions of chewing and deglutition, to recognize the most accepted food consistency, and to evaluate the presence of compensatory maneuvers during deglutition, in children and adolescents with Möbius syndrome. METHODS: The subjects were eight children (three female and five male), with ages ranging from five to 15 years, diagnosed with Möbius syndrome. The children were evaluated during a feeding situation, where they were offered food with different consistencies: liquid, pasty and solid. Data were registered in an evaluation protocol adapted by Guedes, Shintani and Cabello (2003). RESULTS: Results showed a significant relation between tongue mobility condition and type of mastication. The variables speed of mastication and contraction of the masseter muscle were also dependents (p=0018), since there was no change in the speed of mastication when the contraction of the masseter was present. The cheese bread was the best solid consistency food for evaluation, because it produced a more cohesive and less widespread mass in the oral cavity. CONCLUSIONS: The morphofunctional alterations found in patients with Möbius syndrome contributed to change the dynamics of chewing and swallowing. All subjects in the sample used compensatory maneuvers. Therefore, even in the absence of clinical signs that suggest penetration/aspiration, patients with such diagnosis should be referred to speech therapy, in order to provide adequate conditions for a more pleasant meal