Fibroblast-derived extracellular vesicles as trackable efficient transporters of an experimental nanodrug with fibrotic heart and lung targeting.

The discovery of extracellular vesicles (EVs) as efficient exogenous biotransporters of therapeutic agents into cells across biological membranes is an exciting emerging field. Especially the potential of EVs as targeted delivery systems for diseases with selective treatments, such as fibrosis, whose treatment causes side effects in other organs not involved in the disease. Methods: In this study, we collected embryonic fibroblast-derived EVs from two different centrifugation fractions, 10 K g and 100 K g fractions from a NIH-3T3 cell line loaded with an experimental drug. Mice with fibrotic hearts and lungs were obtained by administration of angiotensin II. We generated fluorescent EVs and bioluminescent drug to observe their accumulation by colocalization of their signals in fibrotic heart and lung. The biodistribution of the drug in various organs was obtained by detecting the Au present in the drug nanostructure. Results: The drug-loaded EVs successfully reduced fibrosis in pathological fibroblasts in vitro, and modified the biodistribution of the experimental drug, enabling it to reach the target organs in vivo. We described the pre-analytical characteristics of EVs related to physical variables, culture and harvesting conditions, crucial for their in vivo application as nanotransporters using a previously validated protein-based antifibrotic drug. The results showed the colocalization of EVs and the experimental drug in vivo and ex vivo and the efficient reduction of fibrosis in vitro. This work demonstrates that 10K-EVs and 100K-EVs derived from fibroblasts can act as effective biotransporters for targeted drug delivery to profibrotic fibroblasts, lungs, or heart. Conclusion: We observed that fibroblast-derived 10K-EVs and 100K-EVs are useful biotransporters encapsulating a new generation drug leading to a reduction of fibrosis in profibrotic fibroblasts in vitro. In addition, drug containing EVs were shown to reach fibrotic heart and lungs in vivo, enhancing free drug biodistribution.

Nanozymes with versatile redox capabilities inspired in metalloenzymes.

Metalloenzymes represent exemplary systems in which an organic scaffold combines with a functional inorganic entity, resulting in excellent redox catalysts. Inspired by these natural hybrid biomolecules, biomolecular templates have garnered significant attention for the controlled synthesis of inorganic nanostructures. These nanostructures ultimately benefit from the protection and colloidal stabilization provided by the biomacromolecule. In this study, we have employed this strategy to prepare nanozymes with redox capabilities, utilizing the versatile catalytic profile of Pt-loaded nanomaterials. Thus, we have investigated protein-templated Pt-based nanoclusters of different sizes and compositions, which exhibit remarkable oxidase, catalase, and reductase-like activities. The interplay between the composition and catalytic activity highlighted the size of the nanocluster as the most prominent factor in determining the performance of the nanozymes. Additionally, we have demonstrated the use of protein-templated nanozymes as potential co-catalysts in combination with enzymes for coupled reactions, under both sequential and concurrent one-pot conditions. This study provides valuable insights into nanozyme design and its wide range of applications in the design of complex catalytic systems.

Probing the Cellular Fate of the Protein Corona around Nanoparticles with Nanofocused X-ray Fluorescence Imaging.

X-ray fluorescence imaging (XRF-imaging) with subcellular resolution is used to study the intracellular integrity of a protein corona that was pre-formed around gold nanoparticles (AuNP). Artificial proteins engineered to obtain Gd coordination for detection by XRF-imaging were used to form the corona. Indications about the degradation of this protein corona at a cellular and subcellular level can be observed by following the Au and Gd quantities in a time and spatial-dependent manner. The extended acquisition times necessary for capturing individual XRF-imaging cell images result in relatively small sample populations, stressing the need for faster image acquisition strategies in future XRF-imaging-based studies to deal with the inherent variability between cells. Still, results obtained reveal degradation of the protein corona during cellular trafficking, followed by differential cellular processing for AuNP and Gd-labelled proteins. Overall, this demonstrates that the dynamic degradation of the protein corona can be tracked by XRF-imaging to a certain degree.

Engineered Protein-Driven Synthesis of Tunable Iron Oxide Nanoparticles as T1 and T2 Magnetic Resonance Imaging Contrast Agents.

Iron oxide nanoparticles (IONPs) have become one of the most promising nanomaterials for biomedical applications because of their biocompatibility and physicochemical properties. This study demonstrates the use of protein engineering as a novel approach to design scaffolds for the tunable synthesis of ultrasmall IONPs. Rationally designed proteins, containing different number of metal-coordination sites, were evaluated to control the size and the physicochemical and magnetic properties of a set of protein-stabilized IONPs (Prot-IONPs). Prot-IONPs, synthesized through an optimized coprecipitation approach, presented good T1 and T2 relaxivity values, stability, and biocompatibility, showing potential for magnetic resonance imaging (MRI) applications.

Engineered Repeat Protein Hybrids: The New Horizon for Biologic Medicines and Diagnostic Tools.

ConspectusThe last decades have witnessed unprecedented scientific breakthroughs in all the fields of knowledge, from basic sciences to translational research, resulting in the drastic improvement of the lifespan and overall quality of life. However, despite these great advances, the treatment and diagnosis of some diseases remain a challenge. Inspired by nature, scientists have been exploring biomolecules and their derivatives as novel therapeutic/diagnostic agents. Among biomolecules, proteins raise much interest due to their high versatility, biocompatibility, and biodegradability.Protein binders (binders) are proteins that bind other proteins, in certain cases, inhibiting or modulating their action. Given their therapeutic potential, binders are emerging as the next generation of biopharmaceuticals. The most well-known example of binders are antibodies, and inspired by them researchers have developed alternative binders using protein design approaches. Protein design can be based on naturally occurring proteins in which, by means of rational design or combinatorial approaches, new binding interfaces can be engineered to obtain specific functions or based on de novo proteins emerging from state-of-the-art computational methodologies.Among the novel designed proteins, a class of engineered repeat proteins, the consensus tetratricopeptide repeat (CTPR) proteins, stand out due to their stability and robustness. The CTPR unit is a helix-turn-helix motif constituted of 34 amino acids, of which only 8 are essential to ensure correct folding of the structure. The small number of conserved residues of CTPR proteins leaves plenty of freedom for functional mutations, making them a base scaffold that can be easily and reproducibly tailored to endow desired functions to the protein. For example, the introduction of metal-binding residues (e.g., histidines, cysteines) drives the coordination of metal ions and the subsequent formation of nanomaterials. Additionally, the CTPR unit can be conjugated with other peptides/proteins or repeated in tandem to encode larger CTPR proteins with superhelical structures. These properties allow for the design of both binder and nanomaterial-coordination modules as well as their combination within the same molecule, making the CTPR proteins, as we have demonstrated in several recent examples, the ideal platform to develop protein-nanomaterial hybrids. Generally, the fusion of two distinct materials exploits the best properties of each; however, in protein-nanomaterial hybrids, the fusion takes on a new dimension as new properties arise.These hybrids have ushered the use of protein-based nanomaterials as biopharmaceuticals beyond their original therapeutic scope and paved the way for their use as theranostic agents. Despite several reports of protein-stabilized nanomaterials found in the literature, these systems offer limited control in the synthesis and properties of the grown nanomaterials, as the protein acts just as a stabilizing agent with no significant functional contribution. Therefore, the rational design of protein-based nanomaterials as true theranostic agents is still incipient. In this context, CTPR proteins have emerged as promising scaffolds to hold simultaneously therapeutic and diagnostic functions through protein engineering, as it has been recently demonstrated in pioneering in vitro and in vivo examples.

Dual-Crosslinked Dynamic Hydrogel Incorporating {Mo154 } with pH and NIR Responsiveness for Chemo-Photothermal Therapy.

Polyoxometalates are an emerging class of molecular clusters, with well-defined structures and chemical compositions that are produced through simple, low-cost, and highly reproducible methods. In particular, the wheel-shaped cluster {Mo154 } is a promising photothermal agent due to its intervalence charge transfer transitions. However, its toxicity hinders its systemic administration, being the development of a localized delivery system still incipient. Herein, an injectable and self-healing hydrogel of easy preparation and administration is developed, incorporating both {Mo154 } and doxorubicin for synergistic photothermal and chemotherapy applications. The hydrogel is composed of benzylaldehyde functionalized polyethylene glycol, poly(N-isopropylacrylamide) functionalized chitosan and {Mo154 }. The gelation occurs within 60 s at room temperature, and the dual crosslinking by Schiff base and electrostatic interactions generates a dynamic network, which enables self-healing after injection. Moreover, the hydrogel delivers chemotherapeutic drugs, with a release triggered by dual near infra-red (NIR) radiation and pH changes. This stimuli-responsive release system along with the photothermal conversion ability of the hydrogel allows the simultaneous combination of photothermal and chemotherapy. This synergic system efficiently ablates the cancer tumor in vivo with no systemic toxicity. Overall, this work paves the way for the development of novel {Mo154 }-based systems, incorporated in self-healing and injectable hydrogels for dual chemo-photothermal therapy.

Cellulose Nanocrystals/Chitosan-Based Nanosystems: Synthesis, Characterization, and Cellular Uptake on Breast Cancer Cells.

Cellulose nanocrystals (CNCs) are elongated biobased nanostructures with unique characteristics that can be explored as nanosystems in cancer treatment. Herein, the synthesis, characterization, and cellular uptake on folate receptor (FR)-positive breast cancer cells of nanosystems based on CNCs and a chitosan (CS) derivative are investigated. The physical adsorption of the CS derivative, containing a targeting ligand (folic acid, FA) and an imaging agent (fluorescein isothiocyanate, FITC), on the surface of the CNCs was studied as an eco-friendly methodology to functionalize CNCs. The fluorescent CNCs/FA-CS-FITC nanosystems with a rod-like morphology showed good stability in simulated physiological and non-physiological conditions and non-cytotoxicity towards MDA-MB-231 breast cancer cells. These functionalized CNCs presented a concentration-dependent cellular internalization with a 5-fold increase in the fluorescence intensity for the nanosystem with the higher FA content. Furthermore, the exometabolic profile of the MDA-MB-231 cells exposed to the CNCs/FA-CS-FITC nanosystems disclosed a moderate impact on the cells' metabolic activity, limited to decreased choline uptake and increased acetate release, which implies an anti-proliferative effect. The overall results demonstrate that the CNCs/FA-CS-FITC nanosystems, prepared by an eco-friendly approach, have a high affinity towards FR-positive cancer cells and thus might be applied as nanocarriers with imaging properties for active targeted therapy.

Diagnóstico de enfermagem risco de infecção e infecções relacionadas à assistência à saúde em pacientes com aids / Diagnóstico de enfermería Riesgo de Infección e Infecciones Asociadas a la Atención de Salud en pacientes con sida / Nursing diagnosis risk for infection and cross-infections in aids patients

Resumo Objetivo Identificar quais fatores de risco do diagnóstico de enfermagem risco de infecção estão associados a chances maiores de pessoas com HIV/aids hospitalizadas desenvolverem Infecções Relacionadas à Assistência à Saúde. Métodos Estudo caso-controle no qual os casos foram pacientes com aids hospitalizados que apresentaram Infecções Relacionadas à Assistência à Saúde (n = 104) e os controles foram pacientes com aids hospitalizados que não evoluíram para Infecções Relacionadas à Assistência à Saúde (n = 104). Usaram-se o teste qui-quadrado de Pearson e a regressão logística, e calculou-se a odds ratio. Resultados Peristaltismo alterado, tabagismo, nível reduzido de hemoglobina e leucopenia foram significativamente associados com o desfecho estudado. Na regressão logística, a redução na hemoglobina foi considerada um fator preditor da detecção de risco de infecção. Conclusão Os indicadores tabagismo, leucopenia e nível reduzido de hemoglobina foram identificados na regressão como os preditores mais importantes para identificar o risco de infecção em pessoas vivendo com HIV/aids.

Resumen Objetivo Identificar qué factores de riesgo del diagnóstico de enfermería Riesgo de Infección están relacionados con mayores probabilidades de que personas con el VIH/sida hospitalizadas presenten Infecciones Asociadas a la Atención de Salud. Métodos Estudio caso-control, en el cual los casos fueron pacientes con sida hospitalizados que presentaron Infecciones Asociadas a la Atención de Salud (n = 104) y los controles fueron pacientes con sida hospitalizados que no contrajeron Infecciones Asociadas a la Atención de Salud (n = 104). Se utilizó la prueba χ2 de Pearson y la regresión logística y se calculó el odds ratio. Resultados El peristaltismo alterado, el tabaquismo, el nivel reducido de hemoglobina y la leucopenia fueron significativamente asociados al resultado estudiado. En la regresión logística, la reducción de la hemoglobina fue considerada un factor predictor de la detección del riesgo de infección. Conclusión Los indicadores tabaquismo, leucopenia y nivel reducido de hemoglobina fueron identificados en la regresión como los predictores más importantes para identificar el riesgo de infección en personas que viven con el VIH/sida.

Abstract Objective To identify which risk factors of the Nursing Diagnosis Risk of infection are associated with a greater chance of Hospitalized People Living with HIV/AIDS developing Healthcare-associated Infections. Methods This is a case-control study in which the cases were composed by hospitalized AIDS patients who presented Healthcare-associated Infections (n=104) and, the controls by those who did not progress to Healthcare-associated Infections (n=104). The Pearson Chi-square test, Odds Ratio calculations for risk factors and Logistic Regression were used. Results Altered peristalsis, smoking, decreased hemoglobin and leukopenia were significantly associated with the outcome investigated. In logistic regression, the decrease in hemoglobin was considered a predictor factor for the detection of infection risk. Conclusion The indicators smoking, leucopenia and decreased hemoglobin were recognized in the regression as the most important predictors for identifying the risk of infection in People Living with HIV/AIDS.

Latest Advances on Bacterial Cellulose-Based Materials for Wound Healing, Delivery Systems, and Tissue Engineering.

Bacterial cellulose (BC) is a nanocellulose form produced by some nonpathogenic bacteria. BC presents unique physical, chemical, and biological properties that make it a very versatile material and has found application in several fields, namely in food industry, cosmetics, and biomedicine. This review overviews the latest state-of-the-art usage of BC on three important areas of the biomedical field, namely delivery systems, wound dressing and healing materials, and tissue engineering for regenerative medicine. BC will be reviewed as a promising biopolymer for the design and development of innovative materials for the mentioned applications. Overall, BC is shown to be an effective and versatile carrier for delivery systems, a safe and multicustomizable patch or graft for wound dressing and healing applications, and a material that can be further tuned to better adjust for each tissue engineering application, by using different methods.

Aggregation of risk indicators to cardiometabolic and musculoskeletal health in Brazilian adolescents in the periods 2008/09 and 2013/14 / Agregação dos indicadores de risco à saúde cardiometabólica e musculoesquelética em adolescentes brasileiros nos períodos de 2008/09 e 2013/14

Abstract Objective To assess the occurrence of an aggregate risk to cardiometabolic and musculoskeletal health of Brazilian adolescents in the period 2008/09 and 2013/14 and to identify whether there are differences in risk between the genders and in these periods. Methods This was a trend epidemiological study with a quantitative approach, consisting of a voluntary sample of adolescents from 16 Brazilian states. Data were extracted from the database of Brazil Sports Project (Projeto Esporte Brasil). Health-related physical fitness was evaluated based on body mass index, cardiorespiratory fitness, flexibility, and abdominal strength/resistance. Descriptive analysis, chi-squared test, and Poisson log regression were used for the statistical treatment. Results In the years 2008/09, 14.6% of Brazilian youngsters showed an aggregate risk to cardiometabolic health and 17.1% an aggregate risk for musculoskeletal indicators, whereas in 2013/14, the values of the risk indicators were, respectively 40.0% and 22.4%. It was observed that, in the years 2013/14, the risk to the cardiometabolic health of boys was 2.51 times greater than in 2008/09, while for girls, a three-fold increase in risk was observed. Concerning musculoskeletal health, girls showed a 2.21 risk of being in the risk zone in 2013/14 when compared with 2008/09. Conclusion The occurrence of an aggregate risk to the cardiometabolic and musculoskeletal health of Brazilian adolescents increased in the 2008/09 and 2013/14 periods. Regarding gender, an increase in the cardiometabolic and musculoskeletal risk between these periods was observed in girls. As for boys, an increase was observed only in cardiometabolic risk.

Resumo Objetivo Verificar a ocorrência de risco agregado à saúde cardiometabólica e musculoesquelética de adolescentes brasileiros no período de 2008/09 e 2013/14 e identificar se existem diferenças no risco entre os sexos e nesses períodos. Métodos Trata-se de um estudo epidemiológico de tendência com abordagem quantitativa, composto por uma amostra voluntária de adolescentes, de 16 estados brasileiros. Os dados foram extraídos da base de dados do Projeto Esporte Brasil. A aptidão física relacionada a saúde foi avaliada a partir de: índice de massa corporal, aptidão cardiorrespiratória, flexibilidade, e força/resistência abdominal. Para o tratamento estatístico foi utilizado análise descritiva, qui-quadrado e regressão Poisson log. Resultados Nos anos de 2008/09, 14,6% de jovens brasileiros apresentaram risco à saúde cardiometabólica agregada e 17,1% risco agregado dos indicadores musculoesqueléticos. Enquanto em 2013/14, os valores dos indicadores de risco foram, respectivamente 40,0% e 22,4%. Observou-se que nos anos de 2013/14 o risco à saúde cardiometabólica dos meninos era 2,51 vezes maior que em 2008/09. Já para as meninas o aumento desse risco foi de 3 vezes. No que se refere à saúde musculoesquelética, as meninas apresentaram risco de 2,21 de estar na zona de risco em 2013/14 em relação à 2008/09. Conclusão A ocorrência de risco agregado à saúde cardiometabólica e musculoesquelética de adolescentes brasileiros aumentou nos períodos de 2008/09 e 2013/14. Com relação ao sexo houve um aumento no risco cardiometabólico e musculoesquelético nas meninas entre esses períodos. Já para os meninos houve aumento apenas do risco cardiometabólico.

Aggregation of risk indicators to cardiometabolic and musculoskeletal health in Brazilian adolescents in the periods 2008/09 and 2013/14.

OBJECTIVE: To assess the occurrence of an aggregate risk to cardiometabolic and musculoskeletal health of Brazilian adolescents in the period 2008/09 and 2013/14 and to identify whether there are differences in risk between the genders and in these periods. METHODS: This was a trend epidemiological study with a quantitative approach, consisting of a voluntary sample of adolescents from 16 Brazilian states. Data were extracted from the database of Brazil Sports Project (Projeto Esporte Brasil). Health-related physical fitness was evaluated based on body mass index, cardiorespiratory fitness, flexibility, and abdominal strength/resistance. Descriptive analysis, chi-squared test, and Poisson log regression were used for the statistical treatment. RESULTS: In the years 2008/09, 14.6% of Brazilian youngsters showed an aggregate risk to cardiometabolic health and 17.1% an aggregate risk for musculoskeletal indicators, whereas in 2013/14, the values of the risk indicators were, respectively 40.0% and 22.4%. It was observed that, in the years 2013/14, the risk to the cardiometabolic health of boys was 2.51 times greater than in 2008/09, while for girls, a three-fold increase in risk was observed. Concerning musculoskeletal health, girls showed a 2.21 risk of being in the risk zone in 2013/14 when compared with 2008/09. CONCLUSION: The occurrence of an aggregate risk to the cardiometabolic and musculoskeletal health of Brazilian adolescents increased in the 2008/09 and 2013/14 periods. Regarding gender, an increase in the cardiometabolic and musculoskeletal risk between these periods was observed in girls. As for boys, an increase was observed only in cardiometabolic risk.

Métodos anestésicos intervencionistas no tratamento da dor oncológica / Interventional anesthetic methods for the treatment of oncologic pain

Introdução: A dor, sintoma comum em pacientes oncológicos, é mal controlada com a terapia convencional em cerca de 10% a 25% dos casos, levando a uma piora significativa da qualidade de vida. Para esses pacientes, a terapia intervencionista, como bloqueios nervosos, procedimentos neurolíticos ou cordotomia e aquelas que necessitam de tratamento contínuo, como a terapia de infusão, levam a bons resultados no controle da dor. Método: Esse artigo é uma revisão bibliográfica realizada entre os anos de 2010 até 2018 sobre métodos intervencionistas para o tratamento da dor em pacientes oncológicos. Resultados: Bloqueios de nervos periféricos são uma forma de tratar a dor relacionada ao câncer, uma única injeção ou série de injeções de anestésico local com ou sem adjuvantes, com a expectativa de alívio da dor de 2 a 6 semanas. Quase todas as regiões das extremidades superior e inferior e a maioria das áreas da cabeça, pescoço e tronco podem ser anestesiados por bloqueios de nervos periféricos. Neurólise com agentes químicos ou físicos é uma outra opção, utilizada para bloquear nervos espinhais ou cranianos e os plexos autonômicos, oferece alívio da dor por alguns meses. A neurólise química é realizada com álcool de alta concentração ou fenol, já a física é através da thermal radiofrequency lesioning (RFL) ou a crioanalgesia. Conclusão: O uso de medicamentos neuroaxiais, proporcionam redução da dose de analgésicos opioides se comparada a dose via oral necessária para obter analgesia semelhante. Isso reduz os efeitos secundários desagradáveis. Os candidatos à terapia neuroaxial contínua são aqueles com requisitos crescentes de opiáceos, dor de difícil controle ou intolerância ao plano analgésico escolhido.

Introduction: Cancer pain management by conventional therapy is ineffective in about 10-25% of cases, leading to a significant decrease in quality of life. interventional therapy, applying either one-time session of nerve blocks, neurolytic procedures, or cordotomy, or long-term treatments as infusion therapy is able to improve cancer pain management. Methodology: This article is a bibliographic review carried out between 2010 and 2018 on interventional methods for the treatment of pain in cancer patients. Results: Peripheral nerve blocks are a way to treat oncology pain. A single injection or a series of injections of local anesthetic with or without adjuvants, with an expectation of pain relief of 2 to 6 weeks. Almost all regions of the upper and lower extremities and most areas of the head, neck and trunk may be anesthetized by peripheral nerve blocks. Neurolysis with chemical or physical agents is another option used to block spinal or cranial nerves and autonomic plexuses offers pain relief for a few months. Chemical neurolysis is performed with high concentrated alcohol or phenol, yet the physical is usually through thermal radiofrequency lesioning (RFL) or cryoanalgesia. Conclusion: The use of neuroaxial drugs, provide a dose reduction of opioid analgesics use compared to the oral dose required to obtain similar analgesia. This reduces unpleasant side effects. The candidates for continuous neuroaxial therapy are those with increasing requirements for opioids, pain of difficult control or intolerance to the chosen analgesic plan.

Network meta-analysis of probiotics to prevent respiratory infections in children and adolescents.

Probiotics have emerged as a promising intervention for the prevention of respiratory tract infections (RTIs) in children. Assess the effect of probiotics on prevention of RTIs in children and adolescents. MEDLINE, EMBASE, LILACS, SCIELO, CINAHL, SCOPUS, and Web of Science. Key words: "respiratory tract infections" AND probiotics. Randomized controlled trials RCT assessing the effect of probiotics on RTIs in children and adolescents were included. Two reviewers, working independently, to identify studies that met the eligibility criteria. Main and secondary outcomes were RTIs and adverse effects, respectively. Twenty-one trials with 6.603 participants were included. Pairwise meta-analysis suggested that Lactobacillus casei rhamnosus (LCA) was the only effective probiotic to the rate of RTIs compared to placebo (RR0.38; Crl 0.19-0.45). Network analysis showed that the LCA exhibited 54.7% probability of being classified in first, while the probability of Lactobacillus fermentum CECT5716 (LFC) being last in the ranking was 15.3%. LCA showed no better effect compared to other probiotic strains by indirect analysis. This systematic review found a lack of evidence to support the effect of probiotic on the incidence rate of respiratory infections in children and adolescents. Pediatr Pulmonol. 2017;52:833-843. © 2017 Wiley Periodicals, Inc.

Single-tube nested PCR assay with in-house DNA extraction for Mycobacterium tuberculosis detection in blood and urine.

INTRODUCTION: Molecular analyses are auxiliary tools for detecting Koch's bacilli in clinical specimens from patients with suspected tuberculosis (TB). However, there are still no efficient diagnostic tests that combine high sensitivity and specificity and yield rapid results in the detection of TB. This study evaluated single-tube nested polymerase chain reaction (STNPCR) as a molecular diagnostic test with low risk of cross contamination for detecting Mycobacterium tuberculosis in clinical samples. METHODS: Mycobacterium tuberculosis deoxyribonucleic acid (DNA) was detected in blood and urine samples by STNPCR followed by agarose gel electrophoresis. In this system, reaction tubes were not opened between the two stages of PCR (simple and nested). RESULTS: STNPCR demonstrated good accuracy in clinical samples with no cross contamination between microtubes. Sensitivity in blood and urine, analyzed in parallel, was 35%-62% for pulmonary and 41%-72% for extrapulmonary TB. The specificity of STNPCR was 100% in most analyses, depending on the type of clinical sample (blood or urine) and clinical form of disease (pulmonary or extrapulmonary). CONCLUSIONS: STNPCR was effective in detecting TB, especially the extrapulmonary form for which sensitivity was higher, and had the advantage of less invasive sample collection from patients for whom a spontaneous sputum sample was unavailable. With low risk of cross contamination, the STNPCR can be used as an adjunct to conventional methods for diagnosing TB.

Single-tube nested PCR assay with in-house DNA extraction for Mycobacterium tuberculosis detection in blood and urine

Abstract: INTRODUCTION : Molecular analyses are auxiliary tools for detecting Koch's bacilli in clinical specimens from patients with suspected tuberculosis (TB). However, there are still no efficient diagnostic tests that combine high sensitivity and specificity and yield rapid results in the detection of TB. This study evaluated single-tube nested polymerase chain reaction (STNPCR) as a molecular diagnostic test with low risk of cross contamination for detecting Mycobacterium tuberculosis in clinical samples. METHODS: Mycobacterium tuberculosis deoxyribonucleic acid (DNA) was detected in blood and urine samples by STNPCR followed by agarose gel electrophoresis. In this system, reaction tubes were not opened between the two stages of PCR (simple and nested). RESULTS: STNPCR demonstrated good accuracy in clinical samples with no cross contamination between microtubes. Sensitivity in blood and urine, analyzed in parallel, was 35%-62% for pulmonary and 41%-72% for extrapulmonary TB. The specificity of STNPCR was 100% in most analyses, depending on the type of clinical sample (blood or urine) and clinical form of disease (pulmonary or extrapulmonary). CONCLUSIONS: STNPCR was effective in detecting TB, especially the extrapulmonary form for which sensitivity was higher, and had the advantage of less invasive sample collection from patients for whom a spontaneous sputum sample was unavailable. With low risk of cross contamination, the STNPCR can be used as an adjunct to conventional methods for diagnosing TB.

Osteoarticular tuberculosis in an HIV-positive patient: a case report.

The authors report a case of a 38-year-old HIV-positive woman, with subcutaneous nodules on the thoracic region with 3 months of evolution. Clinical, laboratory, and epidemiological features were evaluated and associated with apparent damage to the T11-T12 vertebrae, identification by imaging tests, positivity in a polymerase chain reaction-based test, and reactivity to the Mantoux tuberculin skin test (PPD-RT 23). The patient was diagnosed with osteoarticular tuberculosis and received treatment for a year, and clinical cure was achieved.

Osteoarticular tuberculosis in an HIV-positive patient: a case report

The authors report a case of a 38-year-old HIV-positive woman, with subcutaneous nodules on the thoracic region with 3 months of evolution. Clinical, laboratory, and epidemiological features were evaluated and associated with apparent damage to the T11-T12 vertebrae, identification by imaging tests, positivity in a polymerase chain reaction-based test, and reactivity to the Mantoux tuberculin skin test (PPD-RT 23). The patient was diagnosed with osteoarticular tuberculosis and received treatment for a year, and clinical cure was achieved.