Reanalysis and validation of the transcriptional pleural fluid signature in pleural tuberculosis.

Introduction: Pleural tuberculosis (PlTB), the most common site of extrapulmonary TB, is characterized by a paucibacillary nature and a compartmentalized inflammatory response in the pleural cavity, both of which make diagnosis and management extremely challenging. Although transcriptional signatures for pulmonary TB have already been described, data obtained by using this approach for extrapulmonary tuberculosis and, specifically, for pleural tuberculosis are scarce and heterogeneous. In the present study, a set of candidate genes previously described in pulmonary TB was evaluated to identify and validate a transcriptional signature in clinical samples from a Brazilian cohort of PlTB patients and those with other exudative causes of pleural effusion. Methods: As a first step, target genes were selected by a random forest algorithm with recursive feature elimination (RFE) from public microarray datasets. Then, peripheral blood (PB) and pleural fluid (PF) samples from recruited patients presenting exudative pleural effusion were collected during the thoracentesis procedure. Transcriptional analysis of the selected top 10 genes was performed by quantitative RT-PCR (RT-qPCR). Results: Reanalysis of the public datasets identified a set of candidate genes (CARD17, BHLHE40, FCGR1A, BATF2, STAT1, BTN3A1, ANKRD22, C1QB, GBP2, and SEPTIN4) that demonstrated a global accuracy of 89.5% in discriminating pulmonary TB cases from other respiratory diseases. Our validation cohort consisted of PlTB (n = 35) patients and non-TB (n = 34) ones. The gene expressions of CARD17, GBP2, and C1QB in PF at diagnosis were significantly different between the two (PlTB and non-TB) groups (p < 0.0001). It was observed that the gene expressions of CARD17 and GBP2 were higher in PlTB PF than in non-TB patients. C1QB showed the opposite behavior, being higher in the non-TB PF. After anti-TB therapy, however, GBP2 gene expression was significantly reduced in PlTB patients (p < 0.001). Finally, the accuracy of the three above-cited highlighted genes in the PF was analyzed, showing AUCs of 91%, 90%, and 85%, respectively. GBP2 was above 80% (sensitivity = 0.89/specificity = 0.81), and CARD17 showed significant specificity (Se = 0.69/Sp = 0.95) in its capacity to discriminate the groups. Conclusion: CARD17, GBP2, and C1QB showed promise in discriminating PlTB from other causes of exudative pleural effusion by providing accurate diagnoses, thus accelerating the initiation of anti-TB therapy.

Screening of latent tuberculosis infection among patients with diabetes mellitus from a high-burden area in Brazil.

Although several cohort studies have raised the important association between diabetes mellitus (DM) and latent tuberculosis infection (LTBI), evidences are limited and controversial. Furthermore, it is well documented that the poor glycemic control may exacerbate the risk for active TB. Thus, the monitoring of diabetic patients living in high-incidence areas for TB is an important concern in views of available diagnostic tests for LTBI. In this cross-sectional study, we estimate the association of DM and LTBI among diabetic patients classified as type-1 DM (T1D) or type-2 DM (T2D) living in Rio de Janeiro, RJ, Brazil - considered a high TB burden region of these country. Non-DM volunteers were included as endemic area healthy controls. All participants were screened for DM using glycosylated-hemoglobin (HbA1c) and for LTBI using the QuantiFERON-TB Gold in Tube (QFT-GIT). Demographic, socioeconomic, clinical and laboratorial data were also assessed. Among 553 included participants, 88 (15.9%) had QFT-GIT positive test, of which 18 (20.5%) were non-DM, 30 (34.1%) T1D and 40 (45.4%) T2D. After adjustments for potential baseline confounders, age, self-reported non-white skin color and an active TB case in the family were significantly associated with LTBI among the studied population by using a hierarchical multivariate logistic regression analysis. Additionally, we verified that T2D patients were able to produce significant increased interferon-gamma (IFN-γ) plasma levels in response to Mycobacterium tuberculosis-specific antigens, when compared to non-DM individuals. Altogether, our data showed an increased prevalence of LTBI among DM patients, albeit non-statistically significant, and point out to important independent factors associated with LTBI, which deserve attention in monitoring patients with DM. Moreover, QFT-GIT test seems to be a good tool to screening LTBI in this population, even in a high TB burden area.

Hyporexia and cellular/biochemical characteristics of pleural fluid as predictive variables on a model for pleural tuberculosis diagnosis.

OBJECTIVES: Pleural tuberculosis (PlTB) diagnosis is a challenge due to its paucibacillary nature and to the need of invasive procedures. This study aimed to identify easily available variables and build a predictive model for PlTB diagnosis which may allow earlier and affordable alternative strategy to be used in basic health care units. METHODS: An observational cross-sectional study compared PlTB and non-TB patients followed at a tertiary Brazilian hospital between 2010 and 2018. Unconditional logistic regression analysis was performed and a Decision Tree Classifier (DTC) model was validated and applied in additional PlTB patients with empiric diagnosis. The accuracy (Acc), sensitivity (Se), specificity (Sp), positive and negative predictive values were calculated. RESULTS: From 1,135 TB patients, 160 were considered for analysis (111 confirmed PlTB and 49 unconfirmed PlTB). Indeed, 58 non-TB patients were enrolled as controls. Hyporexia [adjusted odds ratio (aOR) 27.39 (95% CI 6.26 - 119.89)] and cellular/biochemical characteristics on pleural fluid (PF) (polimorphonuclear in two categories: 3-14% aOR 26.22, 95% CI 7.11 - 96.68 and < 3% aOR 28.67, 95% CI 5.51 - 149.25; and protein ≥ 5g/dL aOR 7.24, 95% CI 3.07 - 17.11) were associated with higher risk for TB. The DTC constructed using these variables showed Acc=87.6%, Se=89.2%, Sp=84.5% for PlTB diagnosis and was successfully applied in unconfirmed PlTB patients. CONCLUSION: The DTC model showed an excellent performance for PlTB diagnosis and can be considered as an alternative diagnostic strategy by using clinical patterns in association with PF cellular/biochemical characteristics, which were affordable and easily performed in basic health care units.

Analysis of geometrical tomographic parameters of furcation lesions in periodontitis patients.

OBJECTIVES: This study was aimed to investigate the relationship between geometric parameters of furcation lesions - maximum area of lesion opening (MALO), angle formed between the roots (ABR), lesion volume (LV) and presence and height of infra-osseous defects (IOD) - and the success of therapy with enamel matrix derivative proteins (EMD) in patients with grade C periodontitis, using cone-beam computed tomography (CBCT). METHODS: The study consisted of two groups of patients with grade C periodontitis: control (surgery) (n = 17) and test (surgery + EMD) (n = 17). Images parameters on CBCT were recorded using OnDemand3D and ITK-SNAP software. RESULTS: Pearson's correlation coefficient demonstrated that only IOD was statistically significant in the probing depth PD (P = 0.01), with a moderate positive correlation (R = 0.59). MALO was found to be statistically significant (P = 0.03) in the test group (surgery + EMD), with moderate negative correlation (R = -0.5). CONCLUSION: The presence of infra-osseous defects and height were relevant in relation to the success of the type of treatment addressed in this study.

NEONATAL PAIN: CHARACTERIZATION OF THE PHYSIOTHERAPIST'S PERCEPTION IN THE NEONATAL INTENSIVE CARE UNIT.

OBJECTIVE: To describe the perception of physiotherapists in neonatal units regarding pain, the use of measurement scales and strategies that minimize pain. METHODS: Interviews were conducted with physiotherapists in hospitals with neonatal units between 2013 and 2015 in Rio de Janeiro. The questions concerned the knowledge of the feeling of pain, from its recognition to its care or treatment. The description of the results was done by comparing public and private hospitals (Fisher''s Exact exact Testtest), considering p<0.05 as significant. RESULTS: 27 hospitals were visited. All the professionals interviewed (n=27) stated that the newborns feel pain, with facial expression being the most cited and known sign for pain. 26% of physiotherapists believe that newborns experience pain at the same magnitude as adults. Among the scales, the Neonatal Infant Pain Scale (NIPS) was the most well known, but only 37% of the units had routine pain assessment protocols. IV cannulation and blood collection were the most mentioned procedures as a cause of pain and there was no difference between public and private hospitals. CONCLUSIONS: There is a gap in the knowledge about neonatal pain and how to evaluate it among the participating physiotherapists, with no systematization of care routines involving this assessment.

Neonatal pain: characterization of the physiotherapist's perception in the neonatal intensive care unit / Dor neonatal: caracterização da percepção do fisioterapeuta na unidade de terapia intensiva neonatal

ABSTRACT Objective: To describe the perception of physiotherapists in neonatal units regarding pain, the use of measurement scales and strategies that minimize pain. Methods: Interviews were conducted with physiotherapists in hospitals with neonatal units between 2013 and 2015 in Rio de Janeiro. The questions concerned the knowledge of the feeling of pain, from its recognition to its care or treatment. The description of the results was done by comparing public and private hospitals (Fisher''s Exact exact Testtest), considering p<0.05 as significant. Results: 27 hospitals were visited. All the professionals interviewed (n=27) stated that the newborns feel pain, with facial expression being the most cited and known sign for pain. 26% of physiotherapists believe that newborns experience pain at the same magnitude as adults. Among the scales, the Neonatal Infant Pain Scale (NIPS) was the most well known, but only 37% of the units had routine pain assessment protocols. IV cannulation and blood collection were the most mentioned procedures as a cause of pain and there was no difference between public and private hospitals. Conclusions: There is a gap in the knowledge about neonatal pain and how to evaluate it among the participating physiotherapists, with no systematization of care routines involving this assessment.

RESUMO Objetivo: Descrever a percepção dos fisioterapeutas de unidades neonatais sobre a dor, a utilização de escalas de mensuração e estratégias que a minimizem. Métodos: Entrevistas foram realizadas com chefes ou rotinas de fisioterapia em hospitais com unidades neonatais entre 2013 e 2015, no Rio de Janeiro. As perguntas versaram sobre o conhecimento da sensação dolorosa, desde seu reconhecimento até seu cuidado ou tratamento. Foi realizada a descrição dos resultados, comparando-se os dados dos hospitais públicos com os privados (teste exato de Fisher), considerando-se p<0,05 como significante. Resultados: Vinte e sete hospitais foram visitados. Todos os profissionais entrevistados (n=27) afirmaram que os recém-nascidos sentem dor, sendo a expressão facial o sinal de dor mais conhecido. Do total de fisioterapeutas entrevistados, 26% acreditam que os neonatos sentem dor na mesma magnitude que o adulto. Entre as escalas, a Neonatal Infant Pain Scale (NIPS) era a mais conhecida, e apenas 37% das unidades possuíam protocolos de avaliação da dor na rotina. As coletas e as punções foram os procedimentos mais mencionados como causa de dor, e não houve diferença entre os hospitais públicos e privados. Conclusões: Constatou-se uma lacuna no conhecimento sobre dor neonatal e como avaliá-la entre os fisioterapeutas participantes, com ausência de sistematização de rotinas assistenciais que envolvam essa aferição.

Predominance of Th1 Immune Response in Pleural Effusion of Patients with Tuberculosis among Other Exudative Etiologies.

Pleural tuberculosis (PlTB), a common form of extrapulmonary TB, remains a challenge in the diagnosis among many causes of pleural effusion. We recently reported that the combinatorial analysis of interferon gamma (IFN-γ), IFN-γ-inducible protein 10 (IP-10), and adenosine deaminase (ADA) from the pleural microenvironment was useful to distinguish pleural effusion caused by TB (microbiologically confirmed or not) among other etiologies. In this cross-sectional cohort study, a set of inflammatory mediators was quantified in blood and pleural fluid (PF) from exudative pleural effusion cases, including PlTB (n = 27) and non-PlTB (nTB) (n = 25) patients. The levels of interleukin-2 (IL-2), IL-4, IL-6, IL-10, IL-17A, IFN-γ, tumor necrosis factor (TNF), IP-10, transforming growth factor ß1 (TGF-ß), and ADA were determined using cytometric bead assay, enzyme-linked immunosorbent assay (ELISA), or biochemical tests. IFN-γ, IP-10, TNF, TGF-ß, and ADA quantified in PF showed significantly higher concentrations in PlTB patients than in nTB patients. When blood and PF were compared, significantly higher concentrations of IL-6 and IL-10 in PF were identified in both groups. TGF-ß, solely, showed significantly increased levels in PF and blood from PlTB patients when both clinical specimens were compared to those from nTB patients. Principal-component analysis (PCA) revealed a T helper type 1 (Th1) pattern attributed mainly to higher levels of IP-10, IFN-γ, TGF-ß, and TNF in the pleural cavity, which was distinct between PlTB and nTB. In conclusion, our findings showed a predominantly cellular immune response in PF from TB cases, rather than other causes of exudative effusion commonly considered in the differential diagnosis of PlTB.

IgA and IgG antibody detection of mycobacterial antigens in pleural fluid and serum from pleural tuberculous patients.

BACKGROUND: A previous study demonstrated pleural fluid (PF) IgA immunodominance for the fused MT10.3:MPT64 protein in pleural tuberculosis (PLTB) cases. However, no clue on the role of IgA and IgG against this and other antigens in PF and serum concerning improved diagnosis is available. Thus, the aim of the present study was to validate PF IgA-MT10.3:MPT64 and evaluate PF and serum IgA and IgG reactivity against this protein, its peptides (F2) and single MPT64, MT10.3 and the PPE59 mycobacterial specific antigens. IgA and IgG ELISA were measured against the antigen in PLTB (n = 29) and other non-TB pleurisy (n = 39) patient samples. RESULTS: The immunodominance of PF IgA-MT10.3:MPT64 was confirmed in PLTB (86.2%) followed by PPE59 (62%), while serum IgA-F2 exhibited 51.7% sensitivity. PF and serum IgG-MT10.3:MPT64 led to 65.5 and 51.7% sensitivity, respectively. However, MT10.3 and MPT64 displayed overall lower sensitivity (≤34.5) for both antibodies. All results at 95% fixed specificity. Combinatory results indicated 93.1% sensitivity for PF IgA-MT10.3:MPT64/-PPE59 and IgA/IgG-MT10.3:MPT64 at 92.3% specificity, followed by IgA-MT10.3:MPT64/-MPT64 or /-F2 (89.6%) without jeopardizing specificity (94.9%). The combinatory results of the PF adenosine deaminase test (ADA) and IgA-MT10.3:MPT64/-F2 demonstrated the highest sensitivity (96.6%), with a specificity of 92.3%. CONCLUSIONS: The PF IgA-MT10:MPT64 immune dominance was validated in PLTB, and its combinatory results with PPE59 or MPT64 or F2 antigens as well as with IgG, are reported herein for the first time, improving their potential to assist diagnosis. Combining PF-ADA and IgA-MT10.3:MPT64/-F2 results achieved better accuracy. Moreover, serum IgG, although less accurate, displays potential beyond microbiological tests.

Neutrophil CD64 expression levels in IGRA-positive individuals distinguish latent tuberculosis from active disease.

BACKGROUND: CD64 (FcγR1) is a high-affinity receptor for monomeric IgG1 and IgG3. Circulating neutrophils express very low amounts of CD64 on their surface. OBJECTIVES: Our primary aim was to investigate the utility of neutrophil CD64 surface expression as a biomarker of active pulmonary tuberculosis (TB). We hypothesised that elevated neutrophil CD64 expression in TB infection would be associated with interferon gamma (IFN-γ) as an inducer of CD64 expression. METHODS: The expression level of CD64 per neutrophil (PMN CD64 index) was quantitatively measured with flow cytometry using a Leuko64 kit in samples from patients with TB and latent TB infection (LTBI) as well as healthy controls, as part of a prospective cohort study in Brazil. FINDINGS: The PMN CD64 index in patients with TB was higher than that in healthy controls and LTBI. Receiver operating characteristic curve analyses determined that the PMN CD64 index could discriminate patients with TB from those with LTBI and healthy individuals. PMN CD64 index levels returned to baseline levels after treatment. CONCLUSIONS: The positive regulation of CD64 expression in circulating neutrophils of patients with active TB could represent an additional biomarker for diagnosis of active TB and could be used for monitoring individuals with LTBI before progression of TB disease.

Superiority of Interferon Gamma Assay Over Tuberculin Skin Test for Latent Tuberculosis in Inflammatory Bowel Disease Patients in Brazil.

BACKGROUND AND AIMS: To compare tuberculin skin test (TST) and interferon gamma release assay (IGRA) in the screening of LTBI among patients with inflammatory bowel disease (IBD) in an endemic area for tuberculosis, to evaluate the need for repeating tests during anti-TNFα, therapy, and to check whether the results may be affected by immunosuppression. METHODS: A cross-sectional study of 110 IBD patients and 64 controls was conducted in Rio de Janeiro, Brazil. The TST was administered after the Quantiferon(®)-TB Gold In-tube test was performed. RESULTS: TST and IGRA agreement was poor regarding diagnosis (kappa: control = 0.318; UC = 0.202; and CD = - 0.093), anti-TNFα therapy (kappa: with anti-TNFα = 0.150; w/o anti-TNFα = - 0.123), and immunosuppressive therapy (IST) (kappa: with IS = - 0.088; w/o IS = 0.146). Indeterminate IGRA was reported in four CD patients on IST. Follow-up tests after anti-TNFα identified conversion in 8.62% using TST and 20.0% using IGRA. Considering IGRA as a criterion standard, TST showed low sensitivity (19.05%) and positive predictive value (PPV) (21.05%). LTBI detection remarkably improved when IGRA was added to TST (sensitivity of 80.95% and PPV of 53.13%). Results were particularly relevant among CD patients where rates started from zero to reach sensitivity and PPV of more than 60%. CONCLUSION: IGRA alone was more effective to detect LTBI than TST alone and had an overall remarkable added value as an add-on sequential test, particularly in CD patients. While cost-effectiveness of these strategies remains to be evaluated, IGRA appears to be justified in CD prior to and during anti-TNFα therapy, where tuberculosis is endemic.

Neutrophil CD64 expression levels in IGRA-positive individuals distinguish latent tuberculosis from active disease

BACKGROUND CD64 (FcγR1) is a high-affinity receptor for monomeric IgG1 and IgG3. Circulating neutrophils express very low amounts of CD64 on their surface. OBJECTIVES Our primary aim was to investigate the utility of neutrophil CD64 surface expression as a biomarker of active pulmonary tuberculosis (TB). We hypothesised that elevated neutrophil CD64 expression in TB infection would be associated with interferon gamma (IFN-γ) as an inducer of CD64 expression. METHODS The expression level of CD64 per neutrophil (PMN CD64 index) was quantitatively measured with flow cytometry using a Leuko64 kit in samples from patients with TB and latent TB infection (LTBI) as well as healthy controls, as part of a prospective cohort study in Brazil. FINDINGS The PMN CD64 index in patients with TB was higher than that in healthy controls and LTBI. Receiver operating characteristic curve analyses determined that the PMN CD64 index could discriminate patients with TB from those with LTBI and healthy individuals. PMN CD64 index levels returned to baseline levels after treatment. CONCLUSIONS The positive regulation of CD64 expression in circulating neutrophils of patients with active TB could represent an additional biomarker for diagnosis of active TB and could be used for monitoring individuals with LTBI before progression of TB disease.

Application of Venn's diagram in the diagnosis of pleural tuberculosis using IFN-γ, IP-10 and adenosine deaminase.

BACKGROUND: Pleural tuberculosis (PlTB) is the most common extrapulmonary manifestation of this infectious disease which still presents high mortality rates worldwide. Conventional diagnostic tests for PlTB register multiple limitations, including the lack of sensitivity of microbiological methods on pleural specimens and the need of invasive procedures such as pleural biopsy performance. In this scenario, the search for biological markers on pleural fluid (PF) has been the target of several studies as a strategy to overcome the limitations of PlTB diagnosis. This study aims to evaluate the use either isolated or in combination with adenosine deaminase (ADA), interferon-gamma (IFN-γ), interferon-gamma inducible protein of 10-kD (IP-10) levels on PF in order to guide an accurate anti-TB treatment in microbiologically non-confirmed cases. METHODS AND FINDINGS: Eighty patients presenting pleural effusion under investigation were enrolled in a cross-sectional study conducted at Pedro Ernesto University Hospital, Rio de Janeiro, RJ, Brazil. Peripheral blood (PB) and PF samples collected from all patients were applied to the commercial IFN-γ release assay, QuantiFERON-TB Gold In-Tube, and samples were analyzed for IFN-γ and IP-10 by immunoassays. ADA activity was determined on PF by the colorimetric method. Based on microbiological and histological criteria, patients were categorized as follow: confirmed PlTB (n = 16), non-confirmed PlTB (n = 17) and non-PlTB (n = 47). The Mycobacterium tuberculosis antigen-specific production of IFN-γ and IP-10 on PB or PF did not show significant differences. However, the basal levels of these biomarkers, as well as the ADA activity on PF, were significantly increased in confirmed PlTB in comparison to non-PlTB group. Receiver operating characteristics curves were performed and the best cut-off points of these three biomarkers were estimated. Their either isolated or combined performances (sensitivity [Se], specificity [Sp], positive predictive value [PPV], negative predictive value [NPV] and accuracy [Acc]) were determined and applied to Venn's diagrams among the groups. Based on the confirmed PlTB cases, IFN-γ showed the best performance of them at a cut-off point of 2.33 IU/mL (Se = 93.8% and Sp = 97.9%) followed by ADA at a cut-off of 25.80 IU/L (Se = 100% and Sp = 84.8%) and IP-10 (Cut-point = 4,361.90 pg/mL, Se = 75% and Sp = 82.6%). IFN-γ plus ADA (cut-point: 25.80 IU/L) represent the most accurate biomarker combination (98.4%), showing Se = 93.7%, Sp = 100%, PPV = 100% and NPV = 97.9%. When this analysis was applied in non-confirmed PlTB, 15/17 (88.2%) presented at least two positive biomarkers in combination. CONCLUSION: IFN-γ, IP-10, and ADA in PlTB effusions are significantly higher than in non-PlTB cases. IFN-γ is an excellent rule-in and rule-out test compared to IP-10 and ADA. The combination of IFN-γ and ADA, in a reviewed cut-off point, showed to be particularly useful to clinicians as their positive results combined prompts immediate treatment for TB while both negative results suggest further investigation.

Mapeamento demográfico e caracterização do perfil de assistência fisioterapêutica oferecida nas unidades de terapia intensiva neonatais do Rio de Janeiro (RJ) / Mapeo demográfico y caracterización del perfil de asistencia fisioterapéutica ofrecida en unidades de cuidados intensivos neonatales de Río de Janeiro (RJ), Brasil / Demographic mapping and characterization of physical therapy care profile offered in Neonatal Intensive Care Units in the City of Rio de Janeiro, Brazil

RESUMO Este estudo visa ao mapeamento e a caracterização do perfil da assistência fisioterapêutica oferecida em unidades de terapia intensiva (UTI) neonatais dos hospitais públicos e privados do Rio de Janeiro (RJ). Para isso, foram realizadas entrevistas com os chefes sobre as rotinas dos serviços de fisioterapia dos hospitais do Rio de Janeiro com UTI neonatais entre janeiro de 2013 e janeiro de 2015. As perguntas abordaram distribuição das horas de trabalho, organização e localização institucional, tempo de experiência na área, tipos de técnicas fisioterapêuticas e recursos utilizados. Foram incluídos 27 hospitais (17 públicos e 10 privados) da seguinte forma: 6 hospitais na Zona Sul, 8 na Zona Norte, 8 na Zona Oeste e 5 no Centro e Zona Portuária. O número total de fisioterapeutas integrantes das equipes foi de 141, sendo que 59% deles eram especialistas em terapia intensiva neonatal. No que se refere aos chefes entrevistados, 16 (59%) também eram especialistas e 21 (79%) possuíam mais de cinco anos de experiência. Foram citadas diversas técnicas fisioterapêuticas, como: fisioterapia motora, vibração torácica e reequilíbrio tóraco-abdominal. A partir do exposto, verificou-se que a assistência fisioterapêutica neonatal do Rio de Janeiro não está distribuída uniformemente no território, estando mais concentrada nas regiões Sul e Central. Além disso, falta padronização das rotinas e carga horária, sendo necessária adequação do perfil assistencial para atenção ideal e integral do recém-nascido (RN).

RESUMEN Este estudio visa el mapeo y la caracterización del perfil de la asistencia fisioterapéutica ofrecida en unidades de cuidados intensivos (UCI) neonatales de los hospitales públicos y privados de Río de Janeiro (RJ), Brasil. Para eso, fueron realizadas entrevistas con los jefes a respecto de las rutinas de los servicios de fisioterapia de los hospitales de Río de Janeiro con UCI neonatales entre enero de 2013 y enero de 2015. Las preguntas abordaron a respecto de distribición de las horas de trabajo, organización y localización institucional, tiempo de experiencia en la área, tipos de técnicas fisioterapéuticas y recursos utilizados. Fueron incluidos 27 hospitales (17 públicos y 10 privados) de la seguiente forma: 6 hospitales en Zona Sul, 8 en Zona Norte, 8 en Zona Oeste y 5 en Centro y Zona Portuaria. El número total de fisioterapeutas integrantes de los equipos fue de 141, siendo que 59% de ellos eron expertos en cuidados intensivos neonatales. En lo que se refiere a los jefes entrevistados, 16 (59%) también eron expertos y 21 (79%) poseían más de cinco años de experiencia. Fueron citadas diversas técnicas fisioterapéuticas, como: fisioterapia motora, vibración torácica y reequilibrio tóraco-abdominal. A partir del exposto, se verificó que la asistencia fisioterapéutica neonatal de Río de Janeiro não está distribuída uniformemente en el territorio, estando más concentrada en las regiones Sul y Central. Además, falta padronización de las rutinas y carga horaria, siendo necesaria adecuación del perfil asistencial para atención ideal y integral del neonato.

ABSTRACT This study aims to map and characterize the profile of physical therapy care offered in intensive care units (ICU) of public and private hospitals in the city of Rio de Janeiro. To this end, a cross-sectional study was conducted by interviews with the heads/routine chiefs of physical therapy services of the hospitals in the city of Rio de Janeiro with neonatal ICU (NICU) from January 2013 to January 2015. The questions comprised the distribution of working hours, organization and institutional location, time of experience in the area, types of physical therapy techniques and resources used. Twenty seven hospitals (17 public and 10 private) were studied as follows: 6 hospitals in the South area of the city, 8 in the North, 8 in the West and 5 in the central area and port area. The total number of physical therapists of the teams was 141, and 59% of them were experts in neonatal intensive care. With regard to the heads/routine chiefs, 16 (59%) were specialists in neonatal intensive care and 21 (79%) had more than five years of experience. Various physical therapy techniques were cited as: motor physical therapy, chest vibration and thoracic-abdominal rebalance. According to the results, there was not a single standard regarding routines, protocols, professional organization and training, and techniques used. In addition, it was found that the physical therapy in ICUs was not held full-time, neither provided the three shifts care.

Isolation of aerobic cultivable cellulolytic bacteria from different regions of the gastrointestinal tract of giant land snail Achatina fulica.

The enzymatic hydrolysis of cellulose by cellulases is one of the major limiting steps in the conversion of lignocellulosic biomass to yield bioethanol. To overcome this hindrance, significant efforts are underway to identify novel cellulases. The snail Achatina fulica is a gastropod with high cellulolytic activity, mainly due to the abundance of glycoside hydrolases produced by both the animal and its resident microbiota. In this study, we partially assessed the cellulolytic aerobic bacterial diversity inside the gastrointestinal tract of A. fulica by culture-dependent methods and evaluated the hydrolytic repertoire of the isolates. Forty bacterial isolates were recovered from distinct segments of the snail gut and identified to the genus level by 16S rRNA gene sequence analysis. Additional phenotypic characterization was performed using biochemical tests provided by the Vitek2 identification system. The overall enzymatic repertoire of the isolated strains was investigated by enzymatic plate assays, containing the following substrates: powdered sugarcane bagasse, carboxymethylcellulose (CMC), p-nitrophenyl-ß-D-glucopyranoside (pNPG), p-nitrophenyl-ß-D-cellobioside (pNPC), 4-methylumbelliferyl-ß-D-glucopyranoside (MUG), 4-methylumbelliferyl-ß-D-cellobioside (MUC), and 4-methylumbelliferyl-ß-D-xylopyranoside (MUX). Our results indicate that the snail A. fulica is an attractive source of cultivable bacteria that showed to be valuable resources for the production of different types of biomass-degrading enzymes.

Perfil de inaptidão na triagem clínica e sorológica de candidatos à doação de sangue / Profile of inability in the clinic and serological screening of candidates for blood donation

Objetivo: Caracterizar os candidatos quanto ao gênero e analisar a prevalência dos critérios de inaptidão adotados às doações de sangue, realizadas no Banco de Sangue Santa Maria, em Santa Maria, RS. Métodos: Dados retrospectivos arquivados foram avaliados, analisando-se os critérios de exclusão na triagem clínica e sorológica, no período de janeiro de 2005 até dezembro de 2010. Foram analisados 20.264 doadores. O estudo foi aprovado pelo Comitê de Ética e Pesquisa da Universidade Federal do Rio Grande do Sul-UFRGS, sob número de protocolo 18147. Resultados: Novecentos e setenta e seis (5%) candidatos foram considerados inaptos pela triagem clínica e 19.288 (95%) foram considerados aptos para prosseguirem com os testes laboratoriais. Dos doadores aptos, 941 (5%) foram excluídos na triagem sorológica, totalizando 18.347 bolsas de sangue disponíveis para uso. Predominaram os doadores do sexo masculino (62%) e a principal causa de exclusão, na triagem clínica, foi hipertensão (0,7%). Para a triagem sorológica, a principal causa de exclusão foi a Doença de Chagas (1,5%). Conclusão: Ressaltou-se a importância da triagem clínica, tendo em vista que ela excluiu 5% dos candidatos à doação e a relevância da triagem sorológica ser feita corretamente, evitando que resultados falso-negativos sejam liberados.

Objective: Characterize donors by gender and analyze theprevalence of the inability criteria of donations made at the SantaMaria Blood Bank, located at Santa Maria, Brazil. Methods: First,we searched historical data for exclusion criteria used in clinicaland serological screenings during the period from January, 2005 toJuly, 2010. We evaluated 20,264 blood donors' data for this study.The Ethical Committee of Rio Grande do Sul Federal UniversityUFRGSapproved this study under the protocol number 18147.Results: Nine hundred and seventy-six (5%) candidates wereconsidered unfit by the clinical screening and 19,288 (95%) wereconsidered fit. From the resulting fit population of the clinicalscreening, 941 (5%) were excluded from donating by the serologicalscreening, totaling 18.347 blood donations fit for use. The resultsshows that majority of the donors were male (62%) and the leadingcause of exclusion from donating in the clinical screening washypertension (0.7%). As for the serological screening, the leadingcause for exclusion was Chagas disease (1.5%). Conclusion: Thestudy stress the importance of the clinical screening process, giventhat it excluded 5% of blood donations, which were unfit for use. Wealso notice the relevance of a correctly done serological screening,thus avoiding that false-negative results are released.

Impacto do Diagnóstico de Câncer de Boca em Alunos da Disciplina de Estomatologia do Departamento de Odontologia da PUC - Minas / Oral Cancer Diagnosis Impact on Stomatology Students of the PUC Minas School of Dentistry

Avaliar o impacto do diagnóstico de câncer bucalnos alunos do curso de Odontologia da Pontifícia UniversidadeCatólica de Minas Gerais. Material e Métodos: 64 alunos, deambos os sexos e em formação universitária, cursando o 50e o 60 períodos da disciplina de Estomatologia doDepartamento de Odontologia da Pontifícia UniversidadeCatólica de Minas Gerais responderam a um questionárioespecífico relacionado aos fatores emocionais associadosao atendimento de pacientes com lesões suspeitas demalignidade até o diagnóstico final e a comunicação ao mesmoe aos seus familiares Resultados: Dados mostraram que54,5% dos alunos apontaram o diagnóstico precoce docâncer como o fator relevante para a recuperação dopaciente e 75% responderam que o choro seguido de revoltae desespero são as reações mais difíceis de lidar diante docomunicado do diagnóstico de câncer ao paciente.Entretanto, o resultado aponta presença de algumastransformações subjetivas experimentadas pelos estudantesapós a experiência clínica com o diagnostico de câncer e aimportância de introduzir no meio acadêmico um espaço paradiscussões interdisciplinares e trocas de experiências evivências que as relações humanas no campo da saúdeevocam. Conclusão: Ficou evidente a necessidade dereconhecer nestes futuros profissionais condutas guiadaspelos valores humanos e não apenas por conhecimentostécnicos e terapêuticos, compreendendo a angústia e o pedidode ajuda, aplicando sua sensibilidade para lidar com o ouvire o sentir de uma pessoa fragilizada pela doença e pelotempo que perdeu para chegar ao diagnóstico e pelastentativas frustradas de solução de seu problema...

To evaluate the impact of the diagnosis of oralcancer in students of the School of Dentistry, PontificalCatholic University of Minas Gerais. Material and methods:64 students of both genders and in higher degree, taking the5th and 6th semesters of the discipline of Stomatology, DentistryDepartment, Pontifical Catholic University of Minas Gerais,answered a questionnaire related to specific emotional factorsassociated with care in patients presenting lesionssuspicious of malignancy until the final diagnosis and thereporting to them and to their families. Results: Importantdata showed that 54.5% of the students indicated earlydetection of cancer as a factor relevant to the patient’srecovery and 75% answered that crying followed by revoltand despair are the reactions most difficult to deal before thenotice of a diagnosis of cancer to a patient. However, theresults indicate presence of some subjective changesexperienced by students after the experience with the clinicaldiagnosis of cancer, and emphasize the importance ofintroducing in the academic space interdisciplinarydiscussions and exchanges of experiences that humanrelations in health evoke. Conclusion: It was evidenced theneed to recognize in those future professionals someconducts guided by human values †and not just therapeuticand technical expertise, understanding the distress and callfor help, applying their sensitivity to deal with the hearingand feeling of a person weakened by illness and by the losttime until diagnosis was confirmed as well as by theunsuccessful attempts to solve his/her problem...

Ação inseticida do extrato de Derris amazonica Killip para Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae) / Insecticide action of the extract of Derris amazonica Killip for Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae)

A abundância e o potencial inseticida de Derris amazonica e a necessidade de controle de Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae) na cultura do feijão-caupi (Vigna unguiculata L. Walp) estimularam a realização desta pesquisa, que objetivou avaliar a ação inseticida do extrato de D. amazonica a adultos de C. arcuatus em condições de laboratório. Os bioensaios testaram as vias de intoxicação por ingestão de folhas contaminadas, contato com superfície contaminada e aplicação tópica, com delineamento experimental inteiramente casualizado, com quatro repetições. Os valores de mortalidade e consumo foliar dos insetos foram submetidos à análise de regressão, sendo utilizada a análise de Probit para determinação das CL50, da DL50 e dos TL50. O extrato de D. amazonica, contendo 3,7% de rotenona, foi tóxico para adultos de C. arcuatus via ingestão de folhas contaminadas (CL50=15,14 µL do extrato.mL-1 de água), superfície contaminada (CL50=0,45 µL do extrato.cm-2) e aplicação tópica (DL50=1,44 µL do extrato.g-1 do inseto). Mortalidades de adultos de C. arcuatus superiores a 80% e os menores tempos letais médios foram obtidos na concentração de 5% (v v-1) do extrato em todos os bioensaios. O consumo foliar de adultos de C. arcuatus foi inversamente proporcional a concentração do extrato quando expostos por via de ingestão foliar ou aplicação tópica, sendo inclusive observada inibição da alimentação dos indivíduos. O extrato de D. amazonica é tóxico para C. arcuatus e inibe a alimentação dos insetos a partir da concentração de 1% (v v-1).

The abundance and insecticidal potential of Derris amazonica in addition to need of controlling Cerotoma arcuatus for bean crop stimulated this research. The objective of this work was to evaluate insecticide action of the extract of D. amazonica to adults of C. arcuatus in laboratory conditions. The bioassays were carried out using three distend methodologies: leaf intake, contact in treated surface (filter paper) and topical application. A completed randomized experimental design was used with four replications. Mortality values and leaf consumption of the insects were subjected to regression analyses, being the Probit analyses used to determine of the i.e., LC50, LT50 and LD50. The extract of D. amazonica containing 3.7% of rotenone was toxic to adults C. arcuatus when exposed to treated leaves (LC50 = 15.14 µl.mL-1), treated surface (LC50 = 0.45 µl.cm-2) and subjected to topical exposure (LD50 = 1.44 µl.g-1). In all bioassays the adults mortality was higher than 80% with lower median lethal times obtained with 5% (v.v-1) concentrations of the extract. Leaf consumption by adults C. arcuatus was inversely proportional to the concentration of the extract when exposed by leaf intake or topical application, also being observed inhibition of feeding individuals. The extract of D. amazonica is toxic to C. arcuatus and inhibits the feeding of insects from the concentration of 1% (v v-1).

Ação inseticida do extrato de Derris amazonica Killip para Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae

The abundance and insecticidal potential of Derris amazonica in addition to need of controlling Cerotoma arcuatus for bean crop stimulated this research. The objective of this work was to evaluate insecticide action of the extract of D. amazonica to adults of C. arcuatus in laboratory conditions. The bioassays were carried out using three distend methodologies: leaf intake, contact in treated surface (filter paper) and topical application. A completed randomized experimental design was used with four replications. Mortality values and leaf consumption of the insects were subjected to regression analyses, being the Probit analyses used to determine of the i.e., LC50, LT50 and LD50. The extract of D. amazonica containing 3.7% of rotenone was toxic to adults C. arcuatus when exposed to treated leaves (LC50 = 15.14 µl.mL-1), treated surface (LC50 = 0.45 µl.cm-2) and subjected to topical exposure (LD50 = 1.44 µl.g-1). In all bioassays the adults mortality was higher than 80% with lower median lethal times obtained with 5% (v.v-1) concentrations of the extract. Leaf consumption by adults C. arcuatus was inversely proportional to the concentration of the extract when exposed by leaf intake or topical application, also being observed inhibition of feeding individuals. The extract of D. amazonica is toxic to C. arcuatus and inhibits the feeding of insects from the concentration of 1% (v v-1).

A abundância e o potencial inseticida de Derris amazonica e a necessidade de controle de Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae) na cultura do feijão-caupi (Vigna unguiculata L. Walp) estimularam a realização desta pesquisa, que objetivou avaliar a ação inseticida do extrato de D. amazonica a adultos de C. arcuatus em condições de laboratório. Os bioensaios testaram as vias de intoxicação por ingestão de folhas contaminadas, contato com superfície contaminada e aplicação tópica, com delineamento experimental inteiramente casualizado, com quatro repetições. Os valores de mortalidade e consumo foliar dos insetos foram submetidos à análise de regressão, sendo utilizada a análise de Probit para determinação das CL50, da DL50 e dos TL50. O extrato de D. amazonica, contendo 3,7% de rotenona, foi tóxico para adultos de C. arcuatus via ingestão de folhas contaminadas (CL50=15,14 µL do extrato.mL-1 de água), superfície contaminada (CL50=0,45 µL do extrato.cm-2) e aplicação tópica (DL50=1,44 µL do extrato.g-1 do inseto). Mortalidades de adultos de C. arcuatus superiores a 80% e os menores tempos letais médios foram obtidos na concentração de 5% (v v-1) do extrato em todos os bioensaios. O consumo foliar de adultos de C. arcuatus foi inversamente proporcional a concentração do extrato quando expostos por via de ingestão foliar ou aplicação tópica, sendo inclusive observada inibição da alimentação dos indivíduos. O extrato de D. amazonica é tóxico para C. arcuatus e inibe a alimentação dos insetos a partir da concentração de 1% (v v-1).

Toxicidade de manipueira de mandioca (Manihot esculenta Crantz) e erva-de-rato (Palicourea marcgravii St. Hill) a adultos de Toxoptera citricida Kirkaldy (Homoptera: Aphididae)

The utilization of plant extracts might be an alternative for the control of crop pests. Between the plants with insecticide potential, stands-out: Erva-de-rato (Palicourea marcgravii) and the Cassava, a byproduct of the manipueira (Manihot esculenta),common plants in the amazon region. That work had as main objective , to investigate the insecticide potential of Manihot esculenta (manipueira of cassava) and Palicourea marcgravii (erva-de-rato) on Toxoptera citricida (brown citrus aphid), in experimental conditions. The freeze dryed Manipueira and and Erva-de-rato extracts, were sprayed on citrus plants infected by brown citros aphid. In laboratory, extracts of these two plants were obtained in five concentrations (10mg/ml, 20mg/ml, 30mg/ml, 40mg/ml and 50mg/ml). All the analyzed concentrations (10 to 50mg/ml) caused mortality on T. citricida of above 50%, and the higher concentration of 50mg/ml caused the mortality of all the insects (n = 100%) in application processes contact. The potential of these extracts, as demonstrated in the tests, confirms that they can be an insecticide alternative, for the control of the brown citrus aphid.

A utilização de extratos de plantas pode ser uma alternativa para o controle de pragas. Dentre as plantas com atividade inseticida, destacam-se a erva-de-rato (Palicourea marcgravii) e o subproduto (manipueira) da produção de farinha de mandioca (Manihot esculenta), plantas comuns na região Amazônica. Esse trabalho teve, como principal objetivo, investigar o potencial inseticida da manipueira e do extrato de erva-de-rato sobre Toxoptera citricida (pulgão-preto do citros). Os extratos liofilizados de manipueira e de erva-de-rato foram pulverizados sobre plantas de citros contendo pulgões em cinco concentrações (10mg/ml, 20mg/ml, 30mg/ml, 40mg/ml e 50mg/ml). Todas as concentrações analisadas causaram mortalidade dos pulgões superior a 50%, sendo que a maior concentração causou a mortalidade de todos os insetos. O potencial destes extratos demonstrado no experimento coloca os mesmos como uma alternativa ao uso de inseticidas sintéticos no controle do pulgão-preto dos citros.