Clinical manifestations and immune response to tuberculosis.

Tuberculosis is a far-reaching, high-impact disease. It is among the top ten causes of death worldwide caused by a single infectious agent; 1.6 million tuberculosis-related deaths were reported in 2021 and it has been estimated that a third of the world's population are carriers of the tuberculosis bacillus but do not develop active disease. Several authors have attributed this to hosts' differential immune response in which cellular and humoral components are involved, along with cytokines and chemokines. Ascertaining the relationship between TB development's clinical manifestations and an immune response should increase understanding of tuberculosis pathophysiological and immunological mechanisms and correlating such material with protection against Mycobacterium tuberculosis. Tuberculosis continues to be a major public health problem globally. Mortality rates have not decreased significantly; rather, they are increasing. This review has thus been aimed at deepening knowledge regarding tuberculosis by examining published material related to an immune response against Mycobacterium tuberculosis, mycobacterial evasion mechanisms regarding such response and the relationship between pulmonary and extrapulmonary clinical manifestations induced by this bacterium which are related to inflammation associated with tuberculosis dissemination through different routes.

Peptide-pulsed dendritic cells' immunomodulating effect regarding Mycobacterium tuberculosis growth in macrophages.

Mycobacterium tuberculosis is one of the most successful pathogens affecting humans, being the main cause of tuberculosis. It accounts for most infectious agent-related deaths worldwide; it has been estimated that a third of the world's population are bacillus carriers. This pathogen's evolutionary adaptation is mainly due to its ability to block a host's immune system by preventing it using an effective immune response in cases of active tuberculosis. Peptide-based synthetic vaccines represent an alternative for counteracting tuberculosis; however, although peptide antigens can be identified, they are not recognised by a host's immune system. An approach using dendritic cells as immunomodulating agents for increasing synthetic peptides' antigenic capacity has thus been advanced. Dendritic cells obtained from IL to 4- and GM-CSF-treated peripheral blood mononuclear cells were pulsed with synthetic Mtb protein peptides which have been reported as participating in mycobacteria-host interactions; their amino acid sequences were modified to improve MHC-II coupling and thus increase their recognition by a host's immune system. pMHC-II/TCR interaction triggered a lymphocyte response which controlled Mtb intracellular growth in infected macrophages. This work has been aimed at contributing to understanding dendritic cells' role in Mycobacterium tuberculosis protein peptide antigen presentation, thereby increasing individuals' immune response as a means of controlling the disease.

The ctpF Gene Encoding a Calcium P-Type ATPase of the Plasma Membrane Contributes to Full Virulence of Mycobacterium tuberculosis.

Identification of alternative attenuation targets of Mycobacterium tuberculosis (Mtb) is pivotal for designing new candidates for live attenuated anti-tuberculosis (TB) vaccines. In this context, the CtpF P-type ATPase of Mtb is an interesting target; specifically, this plasma membrane enzyme is involved in calcium transporting and response to oxidative stress. We found that a mutant of MtbH37Rv lacking ctpF expression (MtbΔctpF) displayed impaired proliferation in mouse alveolar macrophages (MH-S) during in vitro infection. Further, the levels of tumor necrosis factor and interferon-gamma in MH-S cells infected with MtbΔctpF were similar to those of cells infected with the parental strain, suggesting preservation of the immunogenic capacity. In addition, BALB/c mice infected with Mtb∆ctpF showed median survival times of 84 days, while mice infected with MtbH37Rv survived 59 days, suggesting reduced virulence of the mutant strain. Interestingly, the expression levels of ctpF in a mouse model of latent TB were significantly higher than in a mouse model of progressive TB, indicating that ctpF is involved in Mtb persistence in the dormancy state. Finally, the possibility of complementary mechanisms that counteract deficiencies in Ca2+ transport mediated by P-type ATPases is suggested. Altogether, our results demonstrate that CtpF could be a potential target for Mtb attenuation.

Regulatory T Cells as an Escape Mechanism to the Immune Response in Taenia crassiceps Infection.

Murine cysticercosis by Taenia crassiceps is a model for human neurocysticercosis. Genetic and/or immune differences may underlie the higher susceptibility to infection in BALB/cAnN with respect to C57BL/6 mice. T regulatory cells (Tregs) could mediate the escape of T. crassiceps from the host immunity. This study is aimed to investigate the role of Tregs in T. crassiceps establishment in susceptible and non-susceptible mouse strains. Treg and effector cells were quantified in lymphoid organs before infection and 5, 30, 90, and 130 days post-infection. The proliferative response post-infection was characterized in vitro. The expression of regulatory and inflammatory molecules was assessed on days 5 and 30 post-infection. Depletion assays were performed to assess Treg functionality. Significantly higher Treg percentages were observed in BALB/cAnN mice, while increased percentages of activated CD127+ cells were found in C57BL/6 mice. The proliferative response was suppressed in susceptible mice, and Treg proliferation occurred only in susceptible mice. Treg-mediated suppression mechanisms may include IL-10 and TGFß secretion, granzyme- and perforin-mediated cytolysis, metabolic disruption, and cell-to-cell contact. Tregs are functional in BALB/cAnN mice. Therefore Tregs could be allowing parasite establishment and survival in susceptible mice but could play a homeostatic role in non-susceptible strains.

Antibodies targeting Mycobacterium tuberculosis peptides inhibit mycobacterial entry to infection target cells.

The humoral immunity regarding tuberculosis can contribute towards controlling the mycobacteria and the disease. Antigens mediating such type of immunity should thus be evaluated for formulating anti-tuberculosis vaccines. The antigen recognition of seven peptides derived from proteins on Mtb H37Rv envelope and a further seven peptides modified from them was evaluated in sera taken from people suffering Mtb infection and others free from it. Peptide sequences' ability to inhibit Mtb entry to human macrophages was determined in vitro and, after isolating peptide-specific IgG antibodies, it was ascertained which ones were exercising such inhibitory function. Aotus were inoculated with the modified peptides for evaluating the activity of the antibodies so produced. Human QTF+ and QTF- sera recognised some of the peptides and inhibited Mtb entry. The same effect was seen with peptide-specific IgG regarding all the native sequences and modified ones. Sera taken from inoculated Aotus was also able to reduce the pathogen's entry. The data showed that some peptides evaluated in this study could induce antibodies able to inhibit the pathogen's entry to human macrophages, i.e. they could represent candidates for part of an anti-tuberculosis vaccine. The methodology used here complements the evaluation of promising antigens for designing effective vaccines.

Análisis de la distribución espacial y temporal de los virus del Dengue (2006-2017), Zika (2015-2017) y Chikungunya (2014-2017) en Colombia / Analysis of the spatial and temporal distribution of the Dengue virus (2006- 2017), Zika (2015-2017) and Chikungunya (2014-2017) in Colombia

Objetivo: describir la distribución espacial y temporal de los virus del dengue, zika y Chikungunya en Colombia e identificar si existe agregación espacial, temporal y espacio-temporal. Métodos: se desarrolló un estudio descriptivo de la distribución espacial y temporal de los virus del Dengue (2006-2017), Zika (2015-2017) y Chikungunya (2014-2017) en Colombia, utilizando los principios de la estadística espacial, específicamente en el análisis exploratorio de datos espaciales. Resultados: se identificaron zonas de Colombia donde se presenta una mayor densidad y prevalencia de casos. A partir de los 1124 municipios analizados para cada evento (casos de Dengue, Zika y Chikungunya), se comprobó con significancia estadística (p<0.05) la existencia de dos conglomerados espacio-temporales, en la zona sur-occidental de la región andina y en la región de la Orinoquia. Conclusiones: Se demostró la existencia de dos conglomerados para los eventos Dengue, Zika y Chikungunya que podría establecerse como zonas de mayor riesgo de co-infección.

Objective: to describe the spatial and temporal distribution of dengue, zika and Chikungunya viruses in Colombia and to identify clusters at spatial, temporal and space-temporal levels. Methods: A descriptive study was developed about the space and time distribution of the Dengue virus (2006-2017), Zika (2015-2017) and Chikungunya (2014-2017) in Colombia, using principles of spatial statistics, namely the spatial data exploratory analysis. Results: Areas of Colombia were identified where there is a higher density and prevalence of cases and were analyzed 1124 municipalities for each event (cases of Dengue, Zika and Chikungunya). Significant clusters (P<0.05) were proven in spatial, temporal and space-temporal analysis, in the south-western zone of the Andean region and in the Orinoquia region. Conclusions: Two conglomerates were confirmed for the Dengue, Zika and Chikungunya events, that could be established as areas of higher risk of co-infection.

Specific Binding Peptides from Rv3632: A Strategy for Blocking Mycobacterium tuberculosis Entry to Target Cells?

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb, i.e., the aetiological agent); the WHO has established this disease as high priority due to its ensuing mortality. Mtb uses a range of mechanisms for preventing its elimination by an infected host; new, viable alternatives for blocking the host-pathogen interaction are thus sought constantly. This article updates our laboratory's systematic search for antigens using bioinformatics tools to clarify the Mtb H37Rv Rv3632 protein's topology and location. This article reports a C-terminal region consisting of peptides 39255 and 39256 (81Thr-Arg114) having high specific binding regarding two infection-related cell lines (A549 and U937); they inhibited mycobacterial entry to U937 cells in a concentration-dependent manner. Rv3632 forms part of the mycobacterial cell envelope, formed by six linear synthetic peptides. Circular dichroism enabled determining the protein's secondary structure. It was also found that peptide 39254 (61Gly-Thr83) was a HABP for alveolar epithelial cells and inhibited mycobacteria entry to these cells regardless of concentration. Sera from active or latent tuberculosis patients did not recognise HABPs 39254 and 39256. These sequences represent a promising approach aiming at their ongoing modification and for including them when designing a multi-epitope, anti-tuberculosis vaccine.

Experimental models used in evaluating anti-tuberculosis vaccines: the latest advances in the field.

INTRODUCTION: Tuberculosis is an infectious disease which is caused by bacilli from the M. tuberculosis complex. The Mycobacterium bovis Bacillus Calmette-Guérin vaccine is currently available as a prophylactic tool for preventing the disease; it has been shown to be efficient in preventing disseminated forms of tuberculosis during early ages; however, its efficiency is limited in areas where individuals have had prior exposure to environmental mycobacteria, and its efficacy decreases with a host's age. AREAS COVERED: Following a comprehensive search of the available literature, this review describes some of the most frequently used animal models, the most frequently used methods for evaluating efficacy in animal models and some in vitro strategies as alternatives for evaluating vaccines. EXPERT OPINION: Identifying the animal models used up to now for evaluating vaccines during their development stages, their characteristics and limitations, as well as knowledge regarding strategies for evaluating promising vaccine candidate efficacy, will ensure more efficient, reliable and reproducible pre-clinical trials. Although much of the knowledge accrued to date concerning vaccine effectiveness against tuberculosis has been based on animal models, it is clear that large questions still need to be resolved and that extrapolation of such efficacy to humans has yet to be achieved.

Mortality rate of patients with cystic fibrosis on the waiting list and within one year after lung transplantation: a survey of Italian CF centers.

BACKGROUND: Cystic Fibrosis (CF) Centers are involved in the decisions regarding the eligibility of CF patients with end-stage lung disease and timing for inclusion on waiting lists (WL) for lung transplantation (LT). There are currently no data on the mortality rates of Italian CF patients on WL and during the first year after LT and we aimed to assess these outcomes by surveying the CF Centers. METHODS: A survey was sent to Italian CF Centers which were requested to report the age at which all CF subjects included on the WL between 2010 and 2014 were included on the list, admitted to either standard or urgent LT, or had died either while on the WL or within the first 3 and 12 months after LT. All outcomes were recorded by December 31, 2015. RESULTS: Two hundred fifty-nine CF subjects were included on the WL during the 5-year study period. The mortality rate during the WL was 19.3% and was not associated with sex, age at inclusion on the WL or standard or urgent access to LT. 159 (61.4%) subjects underwent LT, 46 (28.9%) with urgent procedure. Deaths within the first 3 and 12 months after LT were significantly more prevalent in individuals who underwent urgent LT compared to those with standard LT (p < 0.01). CONCLUSIONS: The mortality of Italian CF patients, included in our survey, was about twice that reported by the National Transplant Center for all LT indications, including CF, during the same time period and despite the introduction of urgent LT. The latter was associated with an unfavorable early outcome compared to standard LT.

Towards designing a synthetic antituberculosis vaccine: The Rv3587c peptide inhibits mycobacterial entry to host cells.

Mycobacterium tuberculosis is considered one of the most successful pathogens in the history of mankind, having caused 1.7 million deaths in 2016. The amount of resistant and extensively resistant strains has increased; BCG has been the only vaccine to be produced in more than 100 years though it is still unable to prevent the disease's most disseminated form in adults; pulmonary tuberculosis. The search is thus still on-going for candidate antigens for an antituberculosis vaccine. This paper reports the use of a logical and rational methodology for finding such antigens, this time as peptides derived from the Rv3587c membrane protein. Bioinformatics tools were used for predicting mycobacterial surface location and Rv3587c protein structure whilst circular dichroism was used for determining its peptides' secondary structure. Receptor-ligand assays identified 4 high activity binding peptides (HABPs) binding specifically to A549 alveolar epithelial cells and U937 monocyte-derived macrophages, covering the region between amino acids 116 and 193. Their capability for inhibiting Mtb H37Rv invasion was evaluated. The recognition of antibodies from individuals suffering active and latent tuberculosis and from healthy individuals was observed in HABPs capable of avoiding mycobacterial entry to host cells. The results showed that 8 HABPs inhibited such invasion, two of them being common for both cell lines: 39265 (155VLAAYVYSLDNKRLWSNLDT173) and 39266 (174APSNETLVKTFSPGEQVTTY192). Peptide 39265 was the least recognised by antibodies from the individuals' sera evaluated in each group. According to the model proposed by FIDIC regarding synthetic vaccine development, peptide 39265 has become a candidate antigen for an antituberculosis vaccine.

Mycobacterium tuberculosis H37Rv LpqG Protein Peptides Can Inhibit Mycobacterial Entry through Specific Interactions.

Mycobacterium tuberculosis is the causative agent of tuberculosis, a disease causing major mortality worldwide. As part of a systematic methodology for studying M. tuberculosis surface proteins which might be involved in host-pathogen interactions, our group found that LpqG surface protein (Rv3623) found in M. tuberculosis complex strains was located on the mycobacterial envelope and that peptide 16661 (21SGCDSHNSGSLGADPRQVTVY40) had high specific binding to U937 monocyte-derived macrophages and inhibited mycobacterial entry to such cells in a concentration-dependent way. A region having high specific binding to A549 alveolar epithelial cells was found which had low mycobacterial entry inhibition. As suggested in previous studies, relevant sequences in the host-pathogen interaction do not induce an immune response and peptides characterised as HABPs are poorly recognised by sera from individuals regardless of whether they have been in contact with M. tuberculosis. Our approach to designing a synthetic, multi-epitope anti-tuberculosis vaccine has been based on identifying sequences involved in different proteins' mycobacteria-target cell interaction and modifying their sequence to improve their immunogenic characteristics, meaning that peptide 16661 sequence should be considered in such design.

Tasa de filtración glomerular y deterioro funcional en una unidad geriátrica de agudos / Glomerular filtration rate and functional decline in an acute geriatric unit

RESUMEN Introducción: una tasa baja de Filtración Glomerular estimada (FGe) se asocia con deterioro funcional. Se sabe poco sobre esa asociación en ancianos hospitalizados. Objetivo: determinar si la disminución de la FGe se asocia a deterioro funcional. Metodología: estudio de cohorte prospectiva, incluyó 1826 pacientes mayores de 60 años hospitalizados en Unidad Geriátrica de Agudos, admitidos entre enero de 2012 y agosto de 2015. El desenlace fue el estado funcional evaluado mediante el Índice de Barthel (IB) en cuatro momentos. La función renal fue estimada según MDRD-4, se agruparon en cuatro categorías según FGe (normal ≥90, leve 60-89, moderado 59-30, severo <30). Se usaron modelos de regresión logística multivariada y procedimiento Glimmix para análisis longitudinales. Resultados: edad promedio 82,3±7,2 años, 51 % eran mujeres. En la regresión logística multivariada, un IB≤60 al ingreso estuvo asociado con edad ≥80 años, género femenino, comorbilidad alta, deterioro social, hipoalbuminemia, anemia, MMSE<19; mientras que la falla renal leve o moderada, disminuyen este riesgo. En el análisis longitudinal, la disminución de IB total durante el seguimiento se asoció con edad ≥80 años, género femenino, deterioro social, estancia hospitalaria ≥15 días, comorbilidad alta, hipoalbuminemia y MMSE <19. La presencia de falla renal leve, moderada o severa se asoció con mayor IB a través del tiempo. Conclusiones: una disminución de FGe está asociada con bajo riesgo de deterioro funcional al ingreso y al seguimiento. Estos hallazgos difieren de reportes previos en la literatura.

SUMMARY Introduction: Low estimated Glomerular filtration rate (eGFR) is associated with functional decline. Little is known on that association in hospitalized elderly. Objective: Determine if low eGFR is associated with functional decline. Methods: Prospective cohort study that included 1826 patients 60 years and older hospitalized in a Geriatric Acute Unit, admitted between January 2012 and August 2015. The outcome was functional status assessed four times by the Barthel Index (BI). Kidney function was estimated by MDRD-4 IDMS and was grouped into four categories according to eGFR (normal ≥90, mild 60-89, moderate 59-30, severe <30). Multivariate logistic regression models and GLIMMIX procedure for longitudinal analyzes were used. Results: Mean age was 82.3±7.2 years, 51 % were women. In multivariate logistic regression, a BI≤60 at admission was associated with age ≥80, female gender, high comorbidity, social deterioration, hypoalbuminemia, anemia, MMSE<19, while the presence of mild or moderate renal failure reduced this risk. In the longitudinal analysis, lower total BI at follow-up was associated with age ≥80, female gender, social deterioration, hospital stay ≥15 days, high comorbidity, hypoalbuminemia, MMSE<19. The presence of mild, moderate or severe renal impairment was associated with higher BI over time. Conclusions: A low eGFR was associated with lower risk for functional decline at admission and overtime. These findings differ from previous reports in the literature.

Quantifying intracellular Mycobacterium tuberculosis: An essential issue for in vitro assays.

Many studies about intracellular microorganisms which are important regarding diseases affecting public health have been focused on the recognition of host-pathogen interactions, thereby ascertaining the mechanisms by which the pathogen invades a cell and makes it become its host. Such knowledge enables understanding the immunological response triggered by these interactions for obtaining useful information for developing vaccines and drugs. Quantitative cell infection assay protocols are indispensable regarding studies involving Mycobacterium tuberculosis, which takes the lives of more than 2 million people worldwide every year; however, sometimes these are limited by the pathogen's slow growth. Concerning such limitation, a detailed review is presented here regarding the different methods for quantifying and differentiating an intracellular pathogen, the importance of mycobacteria aggregate dissociation and multiplicity of infection (MOI) in infection assays. The methods' differences, advantages, and disadvantages are discussed regarding intra and extracellular bacteria (on cell surface) differentiation, current problems are outlined, as are the solutions provided using fluorophores and projections made concerning quantitative infection assays.

Colesterol total y discapacidad en ancianos hospitalizados: más allá de la enfermedad cardiovascular / Total cholesterol and disability in the hospitalised elderly: beyond cardiovascular disease

Resumen Objetivo: determinar si los niveles de colesterol total predicen recuperación en actividades básicas de la vida diaria. Métodos: estudio de cohorte prospectiva que incluyó 806 pacientes ≥ 60 años, hospitalizados en la unidad geriátrica de agudos entre julio de 2013 y agosto de 2015. El desenlace fue desarrollo de discapacidad evaluada mediante el índice de Barthel ≤ 60 en cuatro momentos: 15 días antes del ingreso, al ingreso, al egreso y 30 días después. El colesterol total fue dividido en cuartiles. Se incluyeron variables demográficas, sociales y clínicas. Se usaron los modelos de regresión logística multivariada para predecir discapacidad y mortalidad, junto con el procedimiento Glimmix para análisis longitudinales y para la predicción del índice de Barthel total. Resultados: la edad fue 82,3 ± 7,2. El 52% eran mujeres. En el análisis multivariado la discapacidad estuvo asociada con edad ≥ 85 años, alta comorbilidad, deterioro social, hipoalbuminemia, MMSE < 19 y delirium. Los cuartiles más altos de colesterol total (Q2, Q3, Q4) tuvieron menor riesgo para discapacidad que el cuartil más bajo (Q1). El análisis longitudinal mostró disminución del índice de Barthel en pacientes con estancia hospitalaria > 15 días, hipoalbuminemia, MMSE < 19 y delirium. Los cuartiles más altos de colesterol total alcanzan mayor índice de Barthel a través del tiempo en comparación con el Q1. La mortalidad estuvo asociada con alta comorbilidad, índice de Barthel ≤ 60 al ingreso y MMSE < 19. Los niveles de colesterol total no se asociaron con mortalidad. Conclusiones: el colesterol total en los cuartiles más altos estuvo asociado con mejoría en la recuperación de las actividades básicas de la vida diaria desde 15 días antes del ingreso hospitalario hasta 30 días después de alta, comparado con el Q1.

Abstract Objective: To determine whether the total cholesterol levels predict recovery in basic activities of daily living. Methods: A prospective cohort study was conducted on 806 patients ≥ 60 years-old, admitted into an Acute Geriatric Unit between July 2013 and August 2015. The outcome was development of a disability assessed by a Barthel index ≤ 60 at four different times: 15 days before admission, on admission, at discharge, and 30 days afterwards. The total cholesterol was divided into quartiles. A record was made of the social and clinical demographic variables. Multivariate logistic regression models were used to predict disability and mortality, together with the Glimmix procedure for the longitudinal analysis and for the prediction of the total Barthel index. Results: The mean age of the patients was 82.3 ± 7.2 years, and 52% were women. In the multivariate analysis, disability was associated with an age ≥ 85 years, high comorbidity, social impairment, low plasma albumin, MMSE < 19, and delirium. The highest total cholesterol quartiles (Q2, Q3, Q4) had a lower risk for disability than the lowest quartile (Q1). The longitudinal analysis showed a decrease in the Barthel index in patients with a hospital stay of > 15 days, low plasma albumin, MMSE < 19, and delirium. The highest total cholesterol quartiles reach a higher Barthel index over time compared with the lowest quartile. The mortality was associated with a high comorbidity, a Barthel index ≤ 60 on admission, and MMSE < 19. The total cholesterol levels were not associated with mortality. Conclusions: The total cholesterol in the highest quartiles was associated with an improvement in the recovery of basic activities of daily living from 15 days before hospital admission up to 30 days after discharge compared with the lowest quartile (Q1).

Propuesta para la formación en Medicina Familiar y Comunitaria, desde la percepción, conceptualización y experiencia práctica de los enfoques de Salud Familiar / Proposal for training in Family and Community Medicine, from the perception, conceptualization and practical experience of Family Health approaches / Proposta de formação em Medicina Familiar e Comunitária, a partir da percepção, conceituação e experiência prática de abordagens de Saúde Familiar

Resumen La existencia de un Plan Decenal de Salud Pública para Colombia y la definición de una Política de Atención Integral en Salud concretaron un nuevo Modelo de Atención basado en la Atención Primaria de la Salud. Esto ubicó a la Salud Familiar y Comunitaria como núcleo fundamental y definió un rol al Médico y al Profesional Familiar y Comunitario, que dimensiona los procesos transdisciplinarios y la articulación de acciones individuales y colectivas, con esquemas pedagógicos holísticos transformadores. Este planteamiento generó un análisis y reflexión de la opinión de una muestra del 14,6% de los clasificados como Salubristas por el Observatorio Laboral del Ministerio de Educación quienes, con la respuesta a 13 preguntas enviadas por vía electrónica, libremente plantearon su percepción, conceptualización y experiencia práctica de los enfoques de Salud Familiar y Comunitaria. Con esta medición se definieron los contenidos básicos que se presentan en este documento con un nuevo enfoque transversal en el Pregrado y los Posgrados, de acuerdo con una propuesta pedagógica transformadora y con visión práctica, encontrándose aquí como destacable, la importante relación entre la educación y la salud integral; el concepto que en el 80% de las observaciones se expresó al denominar el poco compromiso de la comunidad y los profesionales de la salud con una "Cultura de la Salud Integralmente Concebida".

Abstract The existence of a Decennial Public Health Plan for Colombia and the definition of a Comprehensive Health Care Policy concretized a new Model of Care based on Primary Health Care. This placed Family and Community Health as a fundamental nucleus and defined a role for the Family and Community Physician and Practitioner, who dimension the transdisciplinary processes and the articulation of individual and collective actions, with transformational holistic pedagogical schemes. This approach generated an analysis and reflection of the opinion of a sample of 14.6% of those classified as Salubristas by the Labor Observatory of the Ministry of Education who, with the answer to 13 questions sent electronically, freely raised their perception, conceptualization and practical experience of Family and Community Health approaches. With this measurement the basic contents that are presented in this document with a new transversal approach in the Undergraduate and Postgraduates were defined, according to a pedagogical proposal transforming and with practical vision, being here like remarkable, the important relation between the education and integral health; the concept that in 80% of the observations was expressed when denominating the little commitment of the community and the professionals of the health with an "Culture of the Health Integrally Conceived".

Resumo A existência de um Plano de Saúde Pública de dez anos para a Colômbia ea definição de uma Política Integral de Saúde resultou em um novo Modelo de Cuidados Baseado em Atenção Primária à Saúde. Isso colocou a Familia e a Saúde Comunitária como um núcleo fundamental e definiu um papel para o Médico e o Profissional Familiar e Comunitário, que dimensiona os processos transdisciplinares e a articulação de ações individuais e coletivas, com esquemas pedagógicos transformadores holísticos. Esta abordagem gerou uma análise e reflexão da opinião de uma amostra de 14,6% das classificadas como Salubristas pelo Observatório do Trabalho do Ministério da Educação que, com a resposta a 13 questões enviadas eletronicamente, aumentou livremente sua percepção, conceituação e experiência prática de abordagens de saúde familiar e comunitária. Com esta medida, os conteúdos básicos apresentados neste documento foram definidos com um novo foco transversal em Estudos de Graduação e Pós-Graduação, de acordo com uma proposta pedagógica transformadora e com uma visão prática, sendo aqui a relação notável entre educação e saúde abrangente; o conceito de que em 80% das observações foi expressado chamando o pequeno compromisso da comunidade e os profissionais de saúde com uma "Cultura de Saúde Integralmente Concebida".

Identifying and characterising PPE7 (Rv0354c) high activity binding peptides and their role in inhibiting cell invasion.

This study was aimed at characterising the PPE7 protein from the PE/PPE protein family. The presence and transcription of the rv0354c gene in the Mycobacterium tuberculosis complex was determined and the subcellular localisation of the PPE7 protein on mycobacterial membrane was confirmed by immunoelectron microscope. Two peptides were identified as having high binding activity (HABPs) and were tested in vitro regarding the invasion of Mycobacterium tuberculosis H37Rv. HABP 39224 inhibited invasion in A549 epithelial cells and U937 macrophages by more than 50%, whilst HABP 39225 inhibited invasion by 40% in U937 cells. HABP 39224, located in the protein's C-terminal region, has a completely conserved amino acid sequence in M. tuberculosis complex species and could be selected as a base peptide when designing a subunit-based, anti-tuberculosis vaccine.

Triazole resistance in Aspergillus fumigatus isolates from patients with cystic fibrosis in Italy.

BACKGROUND: Aspergillus fumigatus is frequently recovered from respiratory secretions of cystic fibrosis (CF) patients. Azole resistance has been increasingly reported. OBJECTIVES: To assess the prevalence of azole resistance in A. fumigatus isolates from patients followed by two CF centers of northern Italy. METHODS: 423 isolates (220 patients) were screened for azole resistance. Resistance was confirmed with the EUCAST method and cyp51A gene sequencing. Microsatellite genotyping was performed and results were compared with those of environmental resistant isolates. RESULTS: No resistance was detected in one center, while 8.2% of the patients of the other center harbored resistant isolates. The TR34/L98H alteration in the cyp51A gene, present in seven cases, resulted associated with poor in-vitro activity of all tested azoles. CONCLUSIONS: The environmental origin of the resistance seems to be probable since azole resistance was found also in naïve patients and an identical microsatellite genotype in clinical and environmental isolates was observed.

Mycobacterium tuberculosis PE9 protein has high activity binding peptides which inhibit target cell invasion.

PE/PPE proteins are involved in several processes during Mycobacterium tuberculosis (Mtb) infection of target cells; studying them is extremely interesting as they are the only ones from the Mycobacterium genus, they abound in pathogenic species such as Mtb and their function remains yet unknown. The PE9 protein (Rv1088) was characterised, the rv1088 gene was identified by PCR in Mtb complex strains and its expression and localisation on mycobacterial surface was confirmed by Western blot and immunoelectron microscopy. Bioinformatics tools were used for predicting PE9 protein structural aspects and experimental study involved the circular dichroism of synthetic peptides. The peptides were tested in binding assays involving U937 and A549 cells; two high activity binding peptides (HABPs) were found for both cell lines (39226-(1)MSYMIATPAALTAAATDIDGI(21) and 39232-(125)YQRHFGTGGQPEFRQHSEHRR(144)), one for U937 (39231-(104)YAGAGRRQRRRRSGDGQWRLRQ(124)) and one for A549 (39230-(83)YGTGVFRRRRGRQTVTAAEHRA(103)). HABP 39232 inhibited mycobacterial entry to A549 cells (∼70%) and U937 cells (∼50%), peptides 39226 and 39231 inhibited entry to U937 cells (∼60% and 80%, respectively) and peptide 39230 inhibited entry to A549 cells (∼60%). This emphasised HABPs' functional importance in recognition between Mtb H37Rv and target cell receptors. These peptide sequences could be involved in invasion and were recognised by the host's immune system, thereby highlighting their use when designing an efficient anti-tuberculosis multiantigenic vaccine.

Cell-Peptide Specific Interaction Can Inhibit Mycobacterium tuberculosis H37Rv Infection.

Studying proteins from the M. tuberculosis H37Rv envelop is important for understanding host-pathogen interaction regarding bacterial infection and survival within a host; such knowledge is indispensable regarding studies aimed at developing drugs or vaccines against tuberculosis, a disease which continues to cause more than one million deaths worldwide every year. The present work presents a study of the Rv3705c protein which has been described as being an outer protein. Several servers and bioinformatics' tools were used for predicting its location on mycobacterial surface and a 3D model of the protein was obtained which was then compared to experimental circular dichroism results for its peptides. PCR assays were used for corroborating rv3705c gene presence and transcription in a laboratory strain and immunoblotting and electron microscopy were used for confirming protein localisation on cell envelop. Receptor-ligand assays revealed two peptides having high specific binding (HABPs); peptide 38485 ((121)DRAFHRVVDRTVGTSGQTTA(140)) bound to both cell lines used as infection target (U937 and A549 epithelial cell line-derived macrophages) and 38488 ((181)RLRENVLLQAKVTQSGNAGP(200)) bound to U937 cells. It was found that peptide 38485 provided significant inhibition regarding mycobacterial entry to both cell lines in in vitro assays. These results led to proposing peptide 38485 as one of the epitopes to be used in future studies aimed at characterising the immune response of functionally important synthetic peptides which could be included in developing a synthetic anti-tuberculosis vaccine.

Mce4F Mycobacterium tuberculosis protein peptides can inhibit invasion of human cell lines.

This work was aimed at studying the Mycobacterium tuberculosis H37Rv Rv3494c protein, taking into account that it belongs to the mammalian cell entry family (mce) which is thought to have important functions in the disease's pathogenesis. The protein was characterized in silico; its presence on mycobacterial surface was confirmed by immunoelectron microscopy. High-activity binding peptides (HABPs) were identified by binding assays with (125)I; their ability to inhibit mycobacterial entry to two cell lines (U937 alveolar macrophages and A549 epithelial cells) was ascertained and their role in bacterial entry was confirmed by fluorescent microsphere internalization assay. This protein's predicted alpha-helix structure was confirmed by circular dichroism of its peptides. All HABPs inhibited mycobacterial entry to cells and that the 38379 peptide ((201)IDQAGPFLQAQIRAGGDIKSY(220)) had high binding ability and inhibited the mycobacterial entry to both cell lines assayed here. Rv3494c peptides 38370 ((21)LSVMAIFYLRLPATFGIGTY(40)), 38373 ((81)HMRLNSGTAIPSNVTATVRSY(100)) and 38379 ((201)IDQAGPFLQAQIRAGGDIKSY(220)) showed to be HABP and inhibited mycobacterial entry to A549 cells and peptide 38382 ((261)RPSFPALAASLANLGRVGVIY(280)) bind to U937 and inhibited the mycobacterial entry to this cell line; all of these sequences play an important role in cell line recognition and invasion, and may thus be considered in the search for prophylactic candidates against tuberculosis.