Recovery of complete genomes of canine parvovirus from clinical samples.

Canine parvovirus (CPV) is a highly pathogenic virus that affects dogs, especially puppies. CPV is believed to have evolved from feline panleukopenia virus (FPV), eventually giving rise to three antigenic types, CPV-2a, 2b, and 2c. CPV-2 is recognized for its resilience in contaminated environments, ease of transmission among dogs, and pathogenicity for puppies. Despite the relevance of the virus, complete genome sequences of CPV available at GenBank, to date, are scarce. In the current study, we have developed a methodology to allow the recovery of complete CPV-2 genomes directly from clinical samples. For this, seven fecal samples from Gurupi, Tocantins, North Brazil, were collected from puppies with clinical signals of viral enteritis, and submitted to viral DNA isolation and amplification. Two multiplex PCR strategies were designed including primers targeting fragments of 400 base pairs (bp) and 1,000 bp along the complete genome. Sequencing was performed with the Nanopore® technology and results obtained with the two approaches were compared. Genome assembly revealed that the 400 bp amplicons generated larger numbers of reads, allowing a more reliable coverage of the whole genome than those attained with primers targeting the larger (1000 bp) amplicons. Nevertheless, both enrichment methodologies were efficient in amplification and sequencing. Viral genome sequences were of high quality and allowed more precise typing and subtyping of viral genomes compared to the commonly employed strategy relying solely on the analysis of the VP2 region, which is limited in scope. The CPV-2 genomes recovered in this study belong to the CPV2a and CPV-2c subtypes, closely related to isolates from the neighboring Amazonian region. In conclusion, the technique reported here may contribute to increase the number of full CPV genomes available, which is essential for understanding the genetic mechanisms underlying the evolution and spread of CPV-2.

Implementation of an innovative model of community nursing for older adults based on Buurtzorg principles: a scoping review protocol.

OBJECTIVE: The objective of this review is to map the international evidence on the implementation of the Buurtzorg model of community nursing practice for the care of older adults. We will describe where and how it has been used, and the challenges and facilitators of implementing this model of care. INTRODUCTION: The challenges of aging have mobilized health systems around the world to replace the current facility- and disease-centered care model with integrated patient-centered care models. The Buurtzorg model provides autonomy to nurses, who, in turn, empower patients in need-based and self-reliant care. INCLUSION CRITERIA: We will consider both published and unpublished studies and reports exploring the process of implementing the Buurtzorg community nursing model for the care of older adults (65 years and older) internationally, in all settings. Concepts of interest will include where the model has been used, how the model has been implemented, and what challenges and facilitators were reported. METHODS: We will implement a three-step search strategy to locate both published and unpublished primary studies, theses, dissertations, book chapters, and text and opinion reports using the following databases: MEDLINE, LILACS, Cochrane CENTRAL, CINAHL, Web of Science, Google Scholar, Embase, Scopus, ProQuest Dissertations and Theses Global, and the official Buurtzorg website. We will present the search strategy in a PRISMA flow diagram. Data will be extracted using Excel spreadsheets and then analyzed narratively. Extracted data will be quantitatively pooled in tables using descriptive statistics to synthesize the characteristics of the reports and sample, followed by a qualitative summary of how the Buurtzorg model has been used, and the challenges and facilitators of implementing this care model.

Human Cytomegalovirus Interleukin 10 Homologs: Facing the Immune System.

Human Cytomegalovirus (HCMV) can cause a variety of health disorders that can lead to death in immunocompromised individuals and neonates. The HCMV lifecycle comprises both a lytic (productive) and a latent (non-productive) phase. HCMV lytic infection occurs in a wide range of terminally differentiated cell types. HCMV latency has been less well-studied, but one characterized site of latency is in precursor cells of the myeloid lineage. All known viral genes are expressed during a lytic infection and a subset of these are also transcribed during latency. The UL111A gene which encodes the viral IL-10, a homolog of the human IL-10, is one of these genes. During infection, different transcript isoforms of UL111A are generated by alternative splicing. The most studied of the UL111A isoforms are cmvIL-10 (also termed the "A" transcript) and LAcmvIL-10 (also termed the "B" transcript), the latter being a well-characterized latency associated transcript. Both isoforms can downregulate MHC class II, however they differ in a number of other immunomodulatory properties, such as the ability to bind the IL10 receptor and induce signaling through STAT3. There are also a number of other isoforms which have been identified which are expressed by differential splicing during lytic infection termed C, D, E, F, and G, although these have been less extensively studied. HCMV uses the viral IL-10 proteins to manipulate the immune system during lytic and latent phases of infection. In this review, we will discuss the literature on the viral IL-10 transcripts identified to date, their encoded proteins and the structures of these proteins as well as the functional properties of all the different isoforms of viral IL-10.

Polymorphisms in interferon pathway genes and risk of Mycobacterium tuberculosis infection in contacts of tuberculosis cases in Brazil.

BACKGROUND: Host genetic polymorphisms may be important in determining susceptibility to Mycobacterium tuberculosis (Mtb) infection, but their role is not fully understood. Detection of microbial DNA and activation of type I interferon (IFN) pathways regulate macrophage responses to Mtb infection. METHODS: We examined whether seven candidate gene SNPs were associated with tuberculin skin test (TST) positivity in close contacts of microbiologically confirmed pulmonary TB patients in Brazil. Independent associations with TST positivity were tested using multivariable logistic regression (using genotypes and clinical variables) and genetic models. RESULTS: Among 482 contacts of 145 TB index cases, 296 contacts were TST positive. Multivariable regression analysis adjusted for population admixture, age, family relatedness, sex and clinical variables related to increased TB risk demonstrated that SNPs in PYHIN1-IFI16-AIM2 rs1101998 (adjusted OR [aOR]: 3.72; 95%CI=1.15-12.0; p=0.028) and in PYHIN1-IFI16-AIM2 rs1633256 (aOR=24.84; 95%CI=2.26-272.95; p=0.009) were associated with TST positivity in a recessive model. Furthermore, an IRF7 polymorphism (rs11246213) was associated with reduced odds of TST positivity in a dominant model (aOR: 0.50, 95%CI: 0.26-0.93; p=0.029). CONCLUSIONS: Polymorphisms in PYHIN1-IFI16-AIM2 rs1633256, rs1101998 and in IRF7 rs11246213 were associated with altered susceptibility to Mtb infection in this Brazilian cohort.

Relationship of anti-tuberculosis drug-induced liver injury and genetic polymorphisms in CYP2E1 and GST

ABSTRACT Setting: Treatment of tuberculosis (TB) can result in Drug-Induced Liver Injury (DILI) since hepatotoxic metabolites are formed during the biotransformation of isoniazid (INH).DILI can be related to the genetic profile of the patient. Single nucleotide polymorphisms in the CYP2E1 gene and GSTM1 and GSTT1 deletion polymorphisms have been associated with adverse events caused by INH. Objective: To characterize the genetic polymorphisms of CYP2E1, GSTT1 and GSTM1 in TB carriers. Design: This is an observational prospective cohort study of 45 patients undergoing treatment of TB. PCR-RFLP and multiplex-PCR were used. Results: The distribution of genotypic frequency in the promoter region (CYP2E1 gene) was: 98% wild genotype and 2% heterozygous. Intronic region: 78% wild genotype; 20% heterozygous and 2% homozygous variant. GST enzyme genes: 24% Null GSTM1 and 22% Null GSTT1. Patients with any variant allele of the CYP2E1 gene were grouped in the statistical analyses. Conclusion: Patients with the CYP2E1 variant genotype or Null GSTT1 showed higher risk of presenting DILI (p = 0.09; OR: 4.57; 95% CI: 0.75-27.6). Individuals with both genotypes had no increased risk compared to individuals with one genotype.

Relationship of anti-tuberculosis drug-induced liver injury and genetic polymorphisms in CYP2E1 and GST.

SETTING: Treatment of tuberculosis (TB) can result in Drug-Induced Liver Injury (DILI) since hepatotoxic metabolites are formed during the biotransformation of isoniazid (INH). DILI can be related to the genetic profile of the patient. Single nucleotide polymorphisms in the CYP2E1 gene and GSTM1 and GSTT1 deletion polymorphisms have been associated with adverse events caused by INH. OBJECTIVE: To characterize the genetic polymorphisms of CYP2E1, GSTT1 and GSTM1 in TB carriers. DESIGN: This is an observational prospective cohort study of 45 patients undergoing treatment of TB. PCR-RFLP and multiplex-PCR were used. RESULTS: The distribution of genotypic frequency in the promoter region (CYP2E1 gene) was: 98% wild genotype and 2% heterozygous. Intronic region: 78% wild genotype; 20% heterozygous and 2% homozygous variant. GST enzyme genes: 24% Null GSTM1 and 22% Null GSTT1. Patients with any variant allele of the CYP2E1 gene were grouped in the statistical analyses. CONCLUSION: Patients with the CYP2E1 variant genotype or Null GSTT1 showed higher risk of presenting DILI (p=0.09; OR: 4.57; 95% CI: 0.75-27.6). Individuals with both genotypes had no increased risk compared to individuals with one genotype.

Tuberculosis-associated anemia is linked to a distinct inflammatory profile that persists after initiation of antitubercular therapy.

Pulmonary tuberculosis (PTB) is associated with chronic inflammation and anemia. How anemia impacts systemic inflammation in PTB patients undergoing antitubercular therapy (ATT) is not fully understood. In the present study, data on several blood biochemical parameters were retrospectively analyzed from 118 PTB patients during the first 60 days of ATT. Multidimensional statistical analyses were employed to perform detailed inflammatory profiling of patients stratified by anemia status prior to treatment. Anemia was defined as hemoglobin levels <12.5 g/dL for female and <13.5 g/dL for male individuals. The findings revealed that most of anemia cases were likely caused by chronic inflammation. A distinct biosignature related to anemia was detected, defined by increased values of uric acid, C-reactive protein, and erythrocyte sedimentation rate. Importantly, anemic patients sustained increased levels of several biochemical markers at day 60 of therapy. Preliminary analysis failed to demonstrate association between persistent inflammation during ATT with frequency of positive sputum cultures at day 60. Thus, TB patients with anemia exhibit a distinct inflammatory profile, which is only partially reverted at day 60 of ATT.

Polymorphisms in TLR4 and TNFA and Risk of Mycobacterium tuberculosis Infection and Development of Active Disease in Contacts of Tuberculosis Cases in Brazil: A Prospective Cohort Study.

BACKGROUND: The role of genetic polymorphisms in latent tuberculosis (TB) infection and progression to active TB is not fully understood. METHODS: We tested the single-nucleotide polymorphisms (SNPs) rs5743708 (TLR2), rs4986791 (TLR4), rs361525 (TNFA), rs2430561 (IFNG) rs1143627 (IL1B) as risk factors for tuberculin skin test (TST) conversion or development of active TB in contacts of active TB cases. Contacts of microbiologically confirmed pulmonary TB cases were initially screened for longitudinal evaluation up to 24 months, with clinical examination and serial TST, between 1998 and 2004 at a referral center in Brazil. Data and biospecimens were collected from 526 individuals who were contacts of 177 active TB index cases. TST conversion was defined as induration ≥5 mm after a negative TST result (0 mm) at baseline or month 4 visit. Independent associations were tested using logistic regression models. RESULTS: Among the 526 contacts, 60 had TST conversion and 44 developed active TB during follow-up. Multivariable regression analysis demonstrated that male sex (odds ratio [OR]: 2.3, 95% confidence interval [CI]: 1.1-4.6), as well as SNPs in TLR4 genes (OR: 62.8, 95% CI: 7.5-525.3) and TNFA (OR: 4.2, 95% CI: 1.9-9.5) were independently associated with TST conversion. Moreover, a positive TST at baseline (OR: 4.7, 95% CI: 2.3-9.7) and SNPs in TLR4 (OR: 6.5, 95% CI: 1.1-36.7) and TNFA (OR: 12.4, 95% CI:5.1-30.1) were independently associated with incident TB. CONCLUSIONS: SNPs in TLR4 and TNFA predicted both TST conversion and active TB among contacts of TB cases in Brazil.

O modelo de remuneração definindo a forma de cuidar: por que premiar a ineficiência no cuidado ao idoso? / The remuneration model defining care: why is inefficiency in elderly care rewarded?

O modelo hegemônico de remuneração dos serviços de saúde em muitos países, tanto em sistemas públicos quanto naqueles orientados ao mercado de planos privados de saúde, ainda é o de fee-for-service. Este se caracteriza, essencialmente, pelo estímulo à competição por usuários e remuneração por quantidade de serviços produzidos (volume). Não basta mudar o modelo de remuneração sem alterar o modelo assistencial e vice-versa, pois os dois são interdependentes. O importante na escolha de um modelo diferenciado de remuneração é que seja adequado ao tipo de assistência executado e ao objetivo que se deseja atingir. Ao longo de anos de aplicação de determinado modelo assistencial associado a um modelo de remuneração, todo um sistema de saúde fica moldado e programado para um fim. Essa é a principal discussão a ser feita atualmente. Alguns dos problemas do sistema de saúde brasileiro, em especial o suplementar, e que afetam primordialmente o idoso são consequência do modelo adotado há décadas. Para dar conta dessa nova e urgente demanda da sociedade, modelos alternativos de remuneração devem ser implementados para romper o círculo vicioso de sucessão de consultas fragmentadas e descontextualizadas da realidade social e de saúde da pessoa idosa, além da produção de procedimentos desconectados do desfecho esperado.

The hegemonic remuneration model in health services in many countries, both in public and private systems is still fee-for-service. This is characterized, essentially, by the stimulus to the competition for doing procedures and remuneration by quantity of services produced (volume). It is not enough to change the remuneration model without changing the care model, as both are interdependent. What is important in choosing a differentiated remuneration model is if it is appropriate to the type of assistance performed and the objective to be achieved. Throughout years of applying a particular care model associated to a compensation model, a whole health system is shaped and programmed to deliver this result. This is the main discussion to be made nowadays. Some of the problems that we have in our health system, especially the supplementary one, and that primarily affect the elderly, are a consequence of the model adopted decades ago. In order to deal with this new and urgent demand from society, alternative remuneration models must be implemented to break with the vicious circle of succession of fragmented health care of the elderly, as well as the production of procedures disconnected from the expected outcome.

Screening for active pulmonary tuberculosis: Development and applicability of artificial neural network models.

Tuberculosis (TB) remains a significant public health challenge, motivated by the diversity of healthcare epidemiological settings, as other factors. Cost-effective screening has substantial importance for TB control, demanding new diagnostic tools. This paper proposes a decision support tool (DST) for screening pulmonary TB (PTB) patients at a secondary clinic. The DST is composed of an adaptive resonance model (iART) for risk group identification (low, medium and high) and a multilayer perceptron (MLP) neural network for classifying patients as active or inactive PTB. Our tool attains an overall sensitivity (SE) and specificity (SP) of 92% (95% CI; 79-97) and 58% (95% CI; 47-68), respectively. SE values for smear-positive and smear-negative patients are 96% (95% CI; 80-99) and 82% (95% CI; 52-95), as well as higher than 83% (95% CI; 43-97) in low and high-risk cases. Even in scenarios with prevalence up to 20%, negative predictive values superior to 95% are obtained. The proposed DST provides a quick and low-cost pretest for presumptive PTB patients, which is useful to guide confirmatory testing and patient management, especially in settings with limited resources in low and middle-incoming countries.

Aging in Brazil: the building of a healthcare model. / Envelhecer no Brasil: a construção de um modelo de cuidado.

The article discusses the development of a health care model for the elderly, seeking to add to the discussion about the aging of the population in the context of a new epidemiological and demographic scenario. Considering that the aging process in Brazil is relatively recent, more relevant social movements have been described in the construction of health policies directed towards the elderly. After an initial description of the main milestones, we present the model of care considered most appropriate for the best care of the elderly. Based on a critical analysis of health care models for the elderly, the article proposes an approach to care for this age group, focusing on health promotion and prevention, in order to avoid overloading the health system. Integrated care models aim to solve the problem of fragmented and poorly coordinated care in current health systems. The more the healthcare professional knows the history of his patient, the better the results; this is how contemporary and resolutive models of care should work, and it is these that are recommended by the most important national and international health agencies. This article is particularly concerned with a care model that is of higher quality, and is more resolutive and cost-effective.

O artigo aborda o desenvolvimento de um modelo de atenção à saúde do idoso, buscando colaborar com a discussão sobre o envelhecimento populacional trazida pela nova realidade epidemiológica e demográfica. Considerando que o processo de envelhecimento no Brasil é relativamente recente, foram descritos movimentos sociais mais relevantes na construção das políticas de saúde voltadas ao idoso. Após a fase descritiva dos marcos, apresentou-se o modelo considerado mais adequado ao melhor cuidado. A partir de uma análise crítica sobre os modelos de atenção à saúde para idosos, o artigo apresenta uma proposta de linha do cuidado para esse segmento, tendo como foco a promoção e a prevenção da saúde, de modo a evitar a sobrecarga do sistema de saúde. Os modelos de cuidados integrados visam resolver o problema dos cuidados fragmentados e mal coordenados nos sistemas de saúde atuais. Quanto mais o profissional conhecer o histórico do seu paciente, melhores serão os resultados; assim devem funcionar os modelos contemporâneos e resolutivos de cuidado recomendados pelos mais importantes organismos nacionais e internacionais de saúde. Um modelo de cuidado de maior qualidade, mais resolutivo e com melhor relação custo-efetividade é a preocupação deste texto.

Envelhecer no Brasil: a construção de um modelo de cuidado / Aging in Brazil: the building of a healthcare model

Resumo O artigo aborda o desenvolvimento de um modelo de atenção à saúde do idoso, buscando colaborar com a discussão sobre o envelhecimento populacional trazida pela nova realidade epidemiológica e demográfica. Considerando que o processo de envelhecimento no Brasil é relativamente recente, foram descritos movimentos sociais mais relevantes na construção das políticas de saúde voltadas ao idoso. Após a fase descritiva dos marcos, apresentou-se o modelo considerado mais adequado ao melhor cuidado. A partir de uma análise crítica sobre os modelos de atenção à saúde para idosos, o artigo apresenta uma proposta de linha do cuidado para esse segmento, tendo como foco a promoção e a prevenção da saúde, de modo a evitar a sobrecarga do sistema de saúde. Os modelos de cuidados integrados visam resolver o problema dos cuidados fragmentados e mal coordenados nos sistemas de saúde atuais. Quanto mais o profissional conhecer o histórico do seu paciente, melhores serão os resultados; assim devem funcionar os modelos contemporâneos e resolutivos de cuidado recomendados pelos mais importantes organismos nacionais e internacionais de saúde. Um modelo de cuidado de maior qualidade, mais resolutivo e com melhor relação custo-efetividade é a preocupação deste texto.

Abstract The article discusses the development of a health care model for the elderly, seeking to add to the discussion about the aging of the population in the context of a new epidemiological and demographic scenario. Considering that the aging process in Brazil is relatively recent, more relevant social movements have been described in the construction of health policies directed towards the elderly. After an initial description of the main milestones, we present the model of care considered most appropriate for the best care of the elderly. Based on a critical analysis of health care models for the elderly, the article proposes an approach to care for this age group, focusing on health promotion and prevention, in order to avoid overloading the health system. Integrated care models aim to solve the problem of fragmented and poorly coordinated care in current health systems. The more the healthcare professional knows the history of his patient, the better the results; this is how contemporary and resolutive models of care should work, and it is these that are recommended by the most important national and international health agencies. This article is particularly concerned with a care model that is of higher quality, and is more resolutive and cost-effective.

Imbalance of NET and Alpha-1-Antitrypsin in Tuberculosis Patients Is Related With Hyper Inflammation and Severe Lung Tissue Damage.

Background: Pulmonary tuberculosis (PTB) can lead to lung tissue damage (LTD) and compromise the pulmonary capacity of TB patients that evolve to severe PTB. The molecular mechanisms involved in LTD during anti-tuberculous treatment (ATT) remain poorly understood. Methods and findings: We evaluated the role of neutrophil extracellular trap (NET) and the occurrence of LTD through chest radiographic images, the microbial load in sputum, and inflammatory serum profile (IL-12p40/p70, IL-8, IL-17A, IL-23, VEGF-A, MMP-1, and -8, galectin-3, citrunillated histone H3-cit-H3, alpha-1-antitrypsin-α1AT, C-reactive protein-CRP and albumin) in a cohort of 82 PTB patients before and after 60 days of ATT. Using univariate analysis, LTD was associated with neutrophilia and increase of several inflammatory proteins involved in the neutrophil-mediated response, being cit-H3 the more related to the event. In the multivariate analysis, neutrophilia and cit-H3 appear as directly related to LTD. The analysis of the ROC curve at day 60 presented AUC of 0.97 (95.0% CI 0.95-1). Interestingly, at day 0 of ATT, these biomarkers demonstrated fine relation with LTD showing an AUC 0.92 (95.0% CI 0.86-0.99). Despite of that, the same molecules have no impact in culture conversion during ATT. Conclusions: Our data revealed that NETs may play a key role in the pathway responsible for non-specific inflammation and tissue destruction in PTB. High level of cit-H3 and low level of α1AT was observed in the serum of severe TB patients, suggesting a breakdown in the intrinsic control of NET-driven tissue damage. These data show a new insight to knowledge TB immunopathogenesis, the role of neutrophil and NET pathway. Likewise, we identified possible biomarkers to screening of PTB patients eligible to adjuvants therapies, as anti-inflammatories and alpha-1-antitrypsin.

A importância da porta de entrada no sistema: o modelo integral de cuidado para o idoso / The importance of the gateway in the system: the integral model of care for the elderly

Resumo A literatura mostra que a prestação de serviços aos idosos deve ser mais eficiente. Usuários necessitam de serviços que integrem atenção primária e demais serviços, além de equipe de saúde qualificada. O plano de cuidados e a gestão são elementos-chave para o sucesso da assistência. Há necessidade de modificar o modelo assistencial, a começar pela integração e coordenação dos serviços. Este texto é baseado em dois experimentos bem-sucedidos de modelos assistenciais para idosos, o da UnATI-UERJ e do Projeto Idoso Bem-Cuidado, iniciativa da Agência Nacional de Saúde Suplementar, que propõe um modelo inovador de atenção. Ambos são compostos por cinco níveis hierarquizados de cuidado: (1) acolhimento, (2) núcleo integrado de cuidado, (3) ambulatório geriátrico, (4) cuidados complexos de curta duração e (5) cuidados longa duração, nos quais se destacam os três primeiros, nas instâncias leves de cuidado. Identificação do risco e integralidade da atenção nos diferentes pontos da rede são o cerne deste modelo. O principal compromisso do projeto é melhorar a qualidade e a coordenação do atendimento desde a porta de entrada do sistema e ao longo do cuidado, com consequente utilização mais adequada dos recursos e melhor resultado para o paciente.

Abstract The literature shows that the provision of services to the elderly should be more efficient. Users need services that integrate primary care and other services, as well as qualified health staff. The care plan and management are key elements for the success of the care. There is a need to modify the healthcare model, starting with the integration and coordination of services. This text is based on two successful healthcare models for the elderly, that of UnATI-UERJ and the Well-Cared Elderly Project, an initiative of the National Supplementary Health Agency that proposes an innovative model of care. Both are comprised of five hierarchical levels of care - (1) care, (2) integrated care center, (3) geriatric outpatient clinic, (4) short-term complex care and (5) long-term care - the first three stand out, in the light instances of care. Risk identification and completeness of attention at different points in the network are at the heart of this model. The main commitment of the project is to improve the quality and coordination of care from the system's entrance door and throughout care, with consequent better use of system resources, both by health professionals and by users and patients.

Murine Cytomegalovirus Spreads by Dendritic Cell Recirculation.

Herpesviruses have coevolved with their hosts over hundreds of millions of years and exploit fundamental features of their biology. Cytomegaloviruses (CMVs) colonize blood-borne myeloid cells, and it has been hypothesized that systemic dissemination arises from infected stem cells in bone marrow. However, poor CMV transfer by stem cell transplantation argues against this being the main reservoir. To identify alternative pathways for CMV spread, we tracked murine CMV (MCMV) colonization after mucosal entry. We show that following intranasal MCMV infection, lung CD11c+ dendritic cells (DC) migrated sequentially to lymph nodes (LN), blood, and then salivary glands. Replication-deficient virus followed the same route, and thus, DC infected peripherally traversed LN to enter the blood. Given that DC are thought to die locally following their arrival and integration into LN, recirculation into blood represents a new pathway. We examined host and viral factors that facilitated this LN traverse. We show that MCMV-infected DC exited LN by a distinct route to lymphocytes, entering high endothelial venules and bypassing the efferent lymph. LN exit required CD44 and the viral M33 chemokine receptor, without which infected DC accumulated in LN and systemic spread was greatly reduced. Taken together, our studies provide the first demonstration of virus-driven DC recirculation. As viruses follow host-defined pathways, high endothelial venules may normally allow DC to pass from LN back into blood.IMPORTANCE Human cytomegalovirus (HCMV) causes devastating disease in the unborn fetus and in the immunocompromised. There is no licensed vaccine, and preventive measures are impeded by our poor understanding of early events in host colonization. HCMV and murine CMV (MCMV) both infect blood-borne myeloid cells. HCMV-infected blood cells are thought to derive from infected bone marrow stem cells. However, infected stem cells have not been visualized in vivo nor shown to produce virus ex vivo, and hematopoietic transplants poorly transfer infection. We show that MCMV-infected dendritic cells in the lungs reach the blood via lymph nodes, surprisingly migrating into high endothelial venules. Dissemination did not require viral replication. It depended on the constitutively active viral chemokine receptor M33 and on the host hyaluronan receptor CD44. Thus, viral chemokine receptors are a possible target to limit systemic CMV infections.

Supplementary Health and aging after 19 years of regulation: where are we now? / A Saúde Suplementar e o envelhecimento após 19 anos de regulação: onde estamos?

Abstract The present article aims to analyze the changes in supplementary health and their effects on the adherence to and maintenance within health plans of the elderly, to demonstrate the current state of the care model offered, and to begin the debate regarding the importance of changing care models and remuneration in the sector. It was observed that turnover in health plans was lower among the elderly than among the non-elderly population, with greater adherence to individual and pre-regulation plans and a very low adherence to dental plans. The elderly were also more representative over the previous year, demonstrating a greater need for permanence in times of economic crisis. Care and cost data point to the urgent need to reformulate the care practice, supported by structures that are practically non-existent in Brazilian supplemental health care today and are not funded in the sector (such as palliative care and care management physicians, among others). AU

Resumo O artigo tem por objetivo analisar as mudanças ocorridas na saúde suplementar e seus reflexos na adesão e manutenção dos idosos nos planos de saúde, demonstrar o estado atual do modelo de assistência oferecido e iniciar o debate sobre a importância da mudança nos modelos assistenciais e de remuneração no setor. Pode-se perceber menor rotatividade dos idosos nos planos de saúde quando comparados à população não idosa, com maior adesão a planos individuais e anteriores à regulamentação; baixíssima adesão a planos odontológicos; e maior representatividade dos idosos no último ano, demonstrando maior necessidade de permanência em momentos de crise econômica. Dados assistenciais e de custo apontam a urgente necessidade de reformulação da prática de cuidado, com apoio de estruturas hoje praticamente não existentes na saúde suplementar brasileira e não financiadas no setor (como cuidados paliativos, médicos gerenciadores do cuidado, entre outras). AU

Detection of tuberculosis drug resistance: a comparison by Mycobacterium tuberculosis MLPA assay versus Genotype®MTBDRplus.

BACKGROUND: To cope with the emergence of multidrug-resistant tuberculosis (MDR-TB), new molecular methods that can routinely be used to screen for a wide range of drug resistance related genetic markers in the Mycobacterium tuberculosis genome are urgently needed. OBJECTIVE: To evaluate the performance of multiplex ligaton-dependent probe amplification (MLPA) against Genotype® MTBDRplus to detect resistance to isoniazid (INHr) and rifampicin (RIFr). METHOD: 96 culture isolates characterised for identification, drug susceptibility testing (DST) and sequencing of rpoB, katG, and inhA genes were evaluated by the MLPA and Genotype®MTBDRplus assays. RESULTS: With sequencing as a reference standard, sensitivity (SE) to detect INHr was 92.8% and 85.7%, and specificity (SP) was 100% and 97.5%, for MLPA and Genotype®MTBDRplus, respectively. In relation to RIFr, SE was 87.5% and 100%, and SP was 100% and 98.8%, respectively. Kappa value was identical between Genotype®MTBDRplus and MLPA compared with the standard DST and sequencing for detection of INHr [0.83 (0.75-0.91)] and RIFr [0.93 (0.88-0.98)]. CONCLUSION: Compared to Genotype®MTBDRplus, MLPA showed similar sensitivity to detect INH and RIF resistance. The results obtained by the MLPA and Genotype®MTBDRplus assays indicate that both molecular tests can be used for the rapid detection of drug-resistant TB with high accuracy. MLPA has the added value of providing information on the circulating M. tuberculosis lineages.