Investigação sobre os métodos de assepsia e equipamentos de proteção individual utilizados por manicures e pedicures / Research on asepsy methods and personal protective equipment used by manicurs and pedicurers / Investigación sobre métodos de asepsia y equipos de protección individual utilizados por manicuras y pedicuras

Introdução: As atividades laborais de manicures e pedicures geram riscos de exposições acidentais aos patógenos vinculados ao sangue como vírus, fungos e bactérias, devido à falta de adesão às medidas de biossegurança, e procedimentos de esterilização acarretando riscos à saúde dos profissionais e clientes. Objetivo:Verificar os conhecimentos das manicures e pedicures sobre os métodos de limpeza e esterilização de materiais, o uso de equipamentos de proteção individual e doenças relacionadas à profissão. Metodologia:Estudo transversal de abordagem quantitativa, realizado entre Outubro de 2017 a Janeiro de 2018, com 22 participantes finais. Incluídos os maiores de 18 anos, ambos os sexos, experiência mínima de um ano e residir em Nova Floresta. A técnica para seleção da amostra foi snowball. Resultados: A limitação de conhecimento sobre os riscos laborais na exposição de agentes infectantes, a baixa adesão no uso de equipamentos de proteção individuale a falta de padronização no processo de descontaminação, limpeza, desinfecção e esterilização dos instrumentais estão associados ao potencial risco de contaminação por agentes infecciosos, e ao acometimento de doenças infectocontagiosas por manicures, pedicures e clientela. Conclusões:Existem fragilidades no conhecimento destes profissionais,a maioria expõe métodos de antissepsia precários, devido a falta de sistematização e padronização dos procedimentos de limpeza, descontaminação dos instrumentais e o uso de equipamentos de proteção individual, além do conhecimento da profissão ter sido adquirido informalmente (AU).

Introduction: The work activities of manicurists and pedicurists generate risks of accidental exposure to pathogens linked to the blood such as viruses, fungi and bacteria, due to the lack of adherence to biosafety measures, and sterilization procedures causing risks to the health of professionals and clients. Objective:To verify the knowledge of manicurists and pedicurists on methods of cleaning and sterilizing materials, the use of personal protective equipment and diseases related to the profession. Methodology: Cross-sectional study with a quantitative approach, carried out between October 2017 and January 2018, with 22 final participants. Included are those over 18 years of age, both sexes, with at least one year's experience and residing in Nova Floresta. The technique for sample selection was snowball.Results:Limited knowledge about occupational risks in the exposure of infectious agents, low adherence to the use of personal protective equipmentand the lack of standardization in the process of decontamination, cleaning, disinfection and sterilizationof instruments are associated with the potential risk of contamination by infectious agents, and the spread of infectious diseases by manicurists, pedicurists and clientele. Conclusions:There are weaknesses in the knowledge of these professionals, most expose precarious antisepsis methods, due to the lack of systematization and standardization of cleaning procedures, decontamination of instruments and the use of personal protective equipment, in addition to the knowledge of the profession having been acquired informally (AU).

Introducción: Las actividades laborales de manicuras y pedicuras generan riesgos de exposiciónaccidental a patógenos ligados a la sangre como virus, hongos y bacterias, debido al incumplimiento de medidas de bioseguridad y procedimientos de esterilización generando riesgos a la salud de profesionales y clientes. Objetivo:Verificar los conocimientos de manicuras y pedicuras sobre métodos de limpieza y esterilización de materiales, el uso de equipos de protección personal y enfermedades relacionadas con la profesión. Metodología:Estudio transversal de abordaje cuantitativo, realizado entre octubre de 2017 y enero de 2018, con 22 participantes finales. Se incluyen los mayores de 18 años, de ambos sexos, con experiencia mínima de un año y vivir en Nova Floresta. La técnica para la selección de la muestra fue la de bola de nieve. Resultados:El conocimiento limitado sobre los riesgos laborales en la exposición de agentes infecciosos, la baja adherencia al uso de equipo de protección personal y la falta de estandarización en el proceso de descontaminación, limpieza, desinfección y esterilización de instrumentos se asocian con el riesgo potencial de contaminación por agentes infecciosos, y la propagación de enfermedades infecciosas entre manicuras, pedicuras y clientela. Conclusiones:Existen debilidades en el conocimiento de estos profesionales, la mayoría expone métodos de antisepsia precarios, debido a la falta de sistematización y estandarización de los procedimientos de limpieza, descontaminación de los instrumentos y el uso de equipo de protección Personal,además de haber adquirido los conocimientos de la profesión informalmente (AU).

Using social media and personality traits to assess software developers' emotional polarity.

Although human factors (e.g., cognitive functions, behaviors and skills, human error models, etc.) are key elements to improve software development productivity and quality, the role of software developers' emotions and their personality traits in software engineering still needs to be studied. A major difficulty is in assessing developers' emotions, leading to the classic problem of having difficulties understanding what cannot be easily measured. Existing approaches to infer emotions, such as facial expressions, self-assessed surveys, and biometric sensors, imply considerable intrusiveness on developers and tend to be used only during normal working periods. This article proposes to assess the feasibility of using social media posts (e.g., developers' posts on Twitter) to accurately determine the polarity of emotions of software developers over extended periods in a non-intrusive manner, allowing the identification of potentially abnormal periods of negative or positive sentiments of developers that may affect software development productivity or software quality. Our results suggested that Twitter data can serve as a valid source for accurately inferring the polarity of emotions. We evaluated 31 combinations of unsupervised lexicon-based techniques using a dataset with 79,029 public posts from Twitter from sixteen software developers, achieving a macro F1-Score of 0.745 and 76.8% of accuracy with the ensemble comprised of SentiStrength, Sentilex-PT, and LIWC2015_PT lexicons. Among other results, we found a statistically significant difference in tweets' polarities posted during working and non-working periods for 31.25% of the participants, suggesting that emotional polarity monitoring outside working hours could also be relevant. We also assessed the Big Five personality traits of the developers and preliminarily used them to ponder the polarities inferences. In this context, Openness, Conscientiousness, and Extraversion were frequently related to neutral and positive posts, while Neuroticism is associated with negative posts. Our results show that the proposed approach is accurate enough to constitute a simple and non-intrusive alternative to existing methods. Tools using this approach can be applied in real software development environments to support software team workers in making decisions to improve the software development process.

SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation.

SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants-Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients' follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance.

Capture ELISA for KPC Detection in Gram-Negative Bacilli: Development and Standardisation.

The detection of KPC-type carbapenemases is necessary for guiding appropriate antibiotic therapy and the implementation of antimicrobial stewardship and infection control measures. Currently, few tests are capable of differentiating carbapenemase types, restricting the lab reports to their presence or not. The aim of this work was to raise antibodies and develop an ELISA test to detect KPC-2 and its D179 mutants. The ELISA-KPC test was designed using rabbit and mouse polyclonal antibodies. Four different protocols were tested to select the bacterial inoculum with the highest sensitivity and specificity rates. The standardisation procedure was performed using 109 previously characterised clinical isolates, showing 100% of sensitivity and 89% of specificity. The ELISA-KPC detected all isolates producing carbapenemases, including KPC variants displaying the ESBL phenotype such as KPC-33 and -66.

Identification of Zika Virus NS1-Derived Peptides with Potential Applications in Serological Tests.

Zika virus (ZIKV), a mosquito-borne pathogen, is an emerging arbovirus associated with sporadic symptomatic cases of great medical concern, particularly among pregnant women and newborns affected with neurological disorders. Serological diagnosis of ZIKV infection is still an unmet challenge due to the co-circulation of the dengue virus, which shares extensive sequence conservation of structural proteins leading to the generation of cross-reactive antibodies. In this study, we aimed to obtain tools for the development of improved serological tests for the detection of ZIKV infection. Polyclonal sera (pAb) and a monoclonal antibody (mAb 2F2) against a recombinant form of the ZIKV nonstructural protein 1 (NS1) allowed the identification of linear peptide epitopes of the NS1 protein. Based on these findings, six chemically synthesized peptides were tested both in dot blot and ELISA assays using convalescent sera collected from ZIKV-infected patients. Two of these peptides specifically detected the presence of ZIKV antibodies and proved to be candidates for the detection of ZIKV-infected subjects. The availability of these tools opens perspectives for the development of NS1-based serological tests with enhanced sensitivity regarding other flaviviruses.

Topical prebiotics/postbiotics and PRURISCORE validation in atopic dermatitis. International study of 396 patients.

Aim: To investigate the efficacy and tolerability of a cream (Rilastil Xerolact PB) containing a mixture of prebiotics and postbiotics, and to validate the PRURISCORE itch scale in the management of atopic dermatitis.Methods: The study is based on 396 subjects of both sexes in three age groups (i.e., infants, children, adults) suffering from mild/moderate Atopic Dermatitis, recruited from 8 European countries and followed for 3 months.Results: The product demonstrated good efficacy combined with good/very good tolerability in all age groups. In particular, SCORAD, PRURISCORE and IGA scores decreased significantly over the course of the study. The PRURISCORE was preferred to VAS by the vast majority of patients.Conclusion: Even though the role of prebiotics and postbiotics was not formally demonstrated since these substances were part of a complex formulation, it can be reasonably stated that prebiotics and postbiotics have safety and standardization features that probiotics do not have. In addition they are authorized by regulatory authorities, whereas topical probiotics are not.

SARS-CoV-2 rapid antigen test based on a new anti-nucleocapsid protein monoclonal antibody: development and real-time validation

SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants—Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients’ follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance.

Capture ELISA for KPC detection in gram-negative bacilli: development and standardisation

The detection of KPC-type carbapenemases is necessary for guiding appropriate antibiotic therapy and the implementation of antimicrobial stewardship and infection control measures. Currently, few tests are capable of differentiating carbapenemase types, restricting the lab reports to their presence or not. The aim of this work was to raise antibodies and develop an ELISA test to detect KPC-2 and its D179 mutants. The ELISA-KPC test was designed using rabbit and mouse polyclonal antibodies. Four different protocols were tested to select the bacterial inoculum with the highest sensitivity and specificity rates. The standardisation procedure was performed using 109 previously characterised clinical isolates, showing 100% of sensitivity and 89% of specificity. The ELISA-KPC detected all isolates producing carbapenemases, including KPC variants displaying the ESBL phenotype such as KPC-33 and -66.

Identification of zika virus ns1-derived peptides with potential applications in serological tests

Zika virus (ZIKV), a mosquito-borne pathogen, is an emerging arbovirus associated with sporadic symptomatic cases of great medical concern, particularly among pregnant women and newborns affected with neurological disorders. Serological diagnosis of ZIKV infection is still an unmet challenge due to the co-circulation of the dengue virus, which shares extensive sequence conservation of structural proteins leading to the generation of cross-reactive antibodies. In this study, we aimed to obtain tools for the development of improved serological tests for the detection of ZIKV infection. Polyclonal sera (pAb) and a monoclonal antibody (mAb 2F2) against a recombinant form of the ZIKV nonstructural protein 1 (NS1) allowed the identification of linear peptide epitopes of the NS1 protein. Based on these findings, six chemically synthesized peptides were tested both in dot blot and ELISA assays using convalescent sera collected from ZIKV-infected patients. Two of these peptides specifically detected the presence of ZIKV antibodies and proved to be candidates for the detection of ZIKV-infected subjects. The availability of these tools opens perspectives for the development of NS1-based serological tests with enhanced sensitivity regarding other flaviviruses.

Recombinant PilS: Cloning, Expression and Biochemical Characterization of a Pil-Fimbriae Subunit.

Pil-fimbriae is a type IV pili member, which is a remarkably versatile component with a wide variety of functions, including motility, attachment to different surfaces, electrical conductance, DNA acquisition, and secretion of a broad range of structurally distinct protein substrates. Despite the previous functional characterization of Pil, more studies are required to understand the regulation of Pil expression and production, since the exact mechanisms involved in these steps are still unknown. Therefore it is extremely important to have a protein with the correct secondary and tertiary structure that will enable an accurate characterization and a specific antisera generation. For this reason, the aim of this work was to generate potential tools for further investigations to comprehend the mechanisms involved in Pil regulation and its role in pathogenic E. coli infections with the obtaining of a precise native-like recombinant PilS and the corresponding antisera. The pilS gene was successfully cloned into an expression vector, and recombinant PilS (rPilS) was efficiently solubilized and purified by metal affinity chromatography. Protein characterization analyses indicated that rPilS presented native-like secondary and tertiary structures after the refolding process. The generated anti-rPilS sera efficiently recognized recombinant and native proteins from atypical enteropathogenic E. coli strains.

Anti-DENV-NS1 monoclonal antibody for the differential histopathological diagnosis of hemorrhagic fever caused by dengue.

Dengue is the most prevalent arboviral disease in humans in tropical and subtropical regions, especially in urban areas, and can cause major epidemics. Although a self-limiting illness, it may sometimes have serious hemorrhagic manifestations, and the outcome of dengue hemorrhagic fever has similar clinical manifestations as in other infections, which could result in death. Therefore, autopsy procedures are required under certain circumstances such as in hemorrhagic fevers, sometimes to confirm or to clarify the diagnosis that may have epidemiological consequences. Normally, the Immunohistochemistry Laboratory of the Pathology Center of Adolfo Lutz Institute receives autopsy samples from different hospitals in Sao Paulo State to confirm a previous diagnosis, especially hemorrhagic fever of infectious etiology. For this diagnosis, we have been using a mouse polyclonal antibody to dengue virus that often does not provide a clear conclusion, because of background staining or no relevant immunostaining, which hampers the histopathological analysis. Accordingly, in the present study, anti-DENV-NS1 monoclonal antibody (4H2) was tested to determine its accuracy in immunohistochemical analysis. Twenty-four autopsy cases of hemorrhagic febrile syndrome showing histopathological alterations compatible with dengue disease were studied: twenty cases were confirmed by RT-PCR for DENV-2 and in four by RT-PCR for yellow fever virus. Samples from autopsied cases of deaths caused by other infectious diseases (two meningitis C and two severe acute respiratory syndrome caused by influenza A H1N1) were included as negative control cases. Positive immunostaining for DENV-NS1 was detected in 16/20 (80%) liver samples and 11/15 (73%) spleen samples from autopsied hemorrhagic dengue patients, whereas the polyclonal antibody detected DENV antigens in 12/20 (60%) liver and in 6/15 (40%) spleen samples from the same cases. Positive results were not obtained with liver biopsy samples from yellow fever or Neisseria meningitides and Flu-A cases. 4H2 mAb recognizes the native protein of the four DENV serotypes in infected cells and did not cross-react with native ZIKV- or CHKV-infected cells by immunohistochemical assay, so it is a useful tool for differential histopathological conclusion of acute febrile hemorrhagic deaths.

In Silico Prediction and Design of Uropathogenic Escherichia coli Alpha-Hemolysin Generate a Soluble and Hemolytic Recombinant Toxin.

The secretion of α-hemolysin by uropathogenic Escherichia coli (UPEC) is commonly associated with the severity of urinary tract infections, which makes it a predictor of poor prognosis among patients. Accordingly, this toxin has become a target for diagnostic tests and therapeutic interventions. However, there are several obstacles associated with the process of α-hemolysin purification, therefore limiting its utilization in scientific investigations. In order to overcome the problems associated with α-hemolysin expression, after in silico prediction, a 20.48 kDa soluble α-hemolysin recombinant denoted rHlyA was constructed. This recombinant is composed by a 182 amino acid sequence localized in the aa542-723 region of the toxin molecule. The antigenic determinants of the rHlyA were estimated by bioinformatics analysis taking into consideration the tertiary form of the toxin, epitope analysis tools, and solubility inference. The results indicated that rHlyA has three antigenic domains localized in the aa555-565, aa600-610, and aa674-717 regions. Functional investigation of rHlyA demonstrated that it has hemolytic activity against sheep red cells, but no cytotoxic effect against epithelial bladder cells. In summary, the results obtained in this study indicate that rHlyA is a soluble recombinant protein that can be used as a tool in studies that aim to understand the mechanisms involved in the hemolytic and cytotoxic activities of α-hemolysin produced by UPEC. In addition, rHlyA can be applied to generate monoclonal and/or polyclonal antibodies that can be utilized in the development of diagnostic tests and therapeutic interventions.

Recombinant PilS: cloning, expression and biochemical characterization of a Pil-Fimbriae subunit

Pil-fimbriae is a type IV pili member, which is a remarkably versatile component with a wide variety of functions, including motility, attachment to different surfaces, electrical conductance, DNA acquisition, and secretion of a broad range of structurally distinct protein substrates. Despite the previous functional characterization of Pil, more studies are required to understand the regulation of Pil expression and production, since the exact mechanisms involved in these steps are still unknown. Therefore it is extremely important to have a protein with the correct secondary and tertiary structure that will enable an accurate characterization and a specific antisera generation. For this reason, the aim of this work was to generate potential tools for further investigations to comprehend the mechanisms involved in Pil regulation and its role in pathogenic E. coli infections with the obtaining of a precise native-like recombinant PilS and the corresponding antisera. The pilS gene was successfully cloned into an expression vector, and recombinant PilS (rPilS) was efficiently solubilized and purified by metal affinity chromatography. Protein characterization analyses indicated that rPilS presented native-like secondary and tertiary structures after the refolding process. The generated anti-rPilS sera efficiently recognized recombinant and native proteins from atypical enteropathogenic E. coli strains.

In silico prediction and design of uropathogenic Escherichia coli alpha-hemolysin generate a soluble and hemolytic recombinant toxin

The secretion of α-hemolysin by uropathogenic Escherichia coli (UPEC) is commonly associated with the severity of urinary tract infections, which makes it a predictor of poor prognosis among patients. Accordingly, this toxin has become a target for diagnostic tests and therapeutic interventions. However, there are several obstacles associated with the process of α-hemolysin purification, therefore limiting its utilization in scientific investigations. In order to overcome the problems associated with α-hemolysin expression, after in silico prediction, a 20.48 kDa soluble α-hemolysin recombinant denoted rHlyA was constructed. This recombinant is composed by a 182 amino acid sequence localized in the aa542–723 region of the toxin molecule. The antigenic determinants of the rHlyA were estimated by bioinformatics analysis taking into consideration the tertiary form of the toxin, epitope analysis tools, and solubility inference. The results indicated that rHlyA has three antigenic domains localized in the aa555–565, aa600–610, and aa674–717 regions. Functional investigation of rHlyA demonstrated that it has hemolytic activity against sheep red cells, but no cytotoxic effect against epithelial bladder cells. In summary, the results obtained in this study indicate that rHlyA is a soluble recombinant protein that can be used as a tool in studies that aim to understand the mechanisms involved in the hemolytic and cytotoxic activities of α-hemolysin produced by UPEC. In addition, rHlyA can be applied to generate monoclonal and/or polyclonal antibodies that can be utilized in the development of diagnostic tests and therapeutic interventions.

Prevalence and factors associated with sepsis and septic shock in oncological patients in intensive therapy / Prevalencia y factores relacionados a la sepsis y choque séptico en pacientes oncológicos en cuidados intensivos / Prevalência e fatores associados à sepse e choque séptico em pacientes oncológicos em terapia intensiva

ABSTRACT Objectives: to analyze factors associated with sepsis and septic shock in cancer patients in the Intensive Care Unit. Methods: cross-sectional, retrospective study with a quantitative approach, with a sample of 239 patients in an oncology hospital. Secondary data from medical records were used. The outcome variable was "presence of sepsis and/or septic shock"; and exposures: sex, length of stay, origin, use of invasive procedures and primary tumor site. Descriptive, bivariate analyzes and multiple logistic regression models were performed. Results: the prevalence of sepsis was 95% CI: 14.7-24.7 and septic shock of 95% CI: 37.7-50.3. In the multiple analysis, sepsis and/or septic shock were associated with hospital stay longer than seven days, being from the Emergency Department, presence of invasive procedures and hematological site. Conclusions: sepsis and/or septic shock in cancer patients were associated with clinical characteristics and health care factors.

RESUMEN Objetivos: analizar factores relacionados a la sepsis y choque séptico en pacientes oncológicos en Unidad de Cuidados Intensivos. Métodos: estudio transversal, retrospectivo, de abordaje cuantitativo, con muestra de 239 pacientes en un hospital oncológico. Utilizados datos secundarios de los prontuarios. La variable-desfecho fue "presencia de sepsis y/o choque séptico"; y las exposiciones: sexo, tiempo de internación, procedencia, uso de procedimientos invasivos y lugar del tumor primario. Realizados análisis descriptivos, bivariadas y modelos de regresión logística múltiple. Resultados: prevalencia de sepsis fue IC95%: 14,7-24,7 y choque séptico de IC95%: 37,7-50,3. En el análisis múltiple, la sepsis y/o choque séptico se relacionaron al tiempo de internación mayor que siete días (OR=2,29 IC95%:1,25-4,2), ser procedente de Urgencia (OR=3,21; IC 95%: 1,57-6,57), presencia de procedimientos invasivos (OR=11,4 IC95%:1,81-72,0) y lugar hematológico (OR=2,22 IC5%: 1,07-4,61). Conclusiones: la sepsis y/o choque séptico en pacientes oncológicos se relacionaron a características clínicas y factores de asistencia de salud.

RESUMO Objetivos: analisar fatores associados à sepse e choque séptico em pacientes oncológicos em Unidade de Terapia Intensiva. Métodos: estudo transversal, retrospectivo, de abordagem quantitativa, com amostra de 239 pacientes em um hospital oncológico. Utilizaram-se dados secundários dos prontuários. A variável-desfecho foi "presença de sepse e/ou choque séptico"; e as exposições: sexo, tempo de internação, procedência, uso de procedimentos invasivos e sítio do tumor primário. Realizaram-se análises descritivas, bivariadas e modelos de regressão logística múltipla. Resultados: a prevalência de sepse foi IC95%: 14,7-24,7 e choque séptico de IC95%: 37,7-50,3. Na análise múltipla, a sepse e/ou choque séptico associaram-se ao tempo de internação maior que sete dias (OR=2,29 IC95%:1,25-4,2), ser procedente da Urgência (OR=3,21; IC 95%: 1,57-6,57), presença de procedimentos invasivos (OR=11,4 IC95%:1,81-72,0) e sítio hematológico (OR=2,22 IC5%: 1,07-4,61). Conclusões: a sepse e/ou choque séptico em pacientes oncológicos se associaram às características clínicas e fatores da assistência à saúde.

Anti-DENV-NS1 monoclonal antibody for the differential histopathological diagnosis of hemorrhagic fever caused by dengue

Dengue is the most prevalent arboviral disease in humans in tropical and subtropical regions, especially in urban areas, and can cause major epidemics. Although a self-limiting illness, it may sometimes have serious hemorrhagic manifestations, and the outcome of dengue hemorrhagic fever has similar clinical manifestations as in other infections, which could result in death. Therefore, autopsy procedures are required under certain circumstances such as in hemorrhagic fevers, sometimes to confirm or to clarify the diagnosis that may have epidemiological consequences. Normally, the Immunohistochemistry Laboratory of the Pathology Center of Adolfo Lutz Institute receives autopsy samples from different hospitals in Sao Paulo State to confirm a previous diagnosis, especially hemorrhagic fever of infectious etiology. For this diagnosis, we have been using a mouse polyclonal antibody to dengue virus that often does not provide a clear conclusion, because of background staining or no relevant immunostaining, which hampers the histopathological analysis. Accordingly, in the present study, anti-DENV-NS1 monoclonal antibody (4H2) was tested to determine its accuracy in immunohistochemical analysis. Twenty-four autopsy cases of hemorrhagic febrile syndrome showing histopathological alterations compatible with dengue disease were studied: twenty cases were confirmed by RT-PCR for DENV-2 and in four by RT-PCR for yellow fever virus. Samples from autopsied cases of deaths caused by other infectious diseases (two meningitis C and two severe acute respiratory syndrome caused by influenza A H1N1) were included as negative control cases. Positive immunostaining for DENV-NS1 was detected in 16/20 (80%) liver samples and 11/15 (73%) spleen samples from autopsied hemorrhagic dengue patients, whereas the polyclonal antibody detected DENV antigens in 12/20 (60%) liver and in 6/15 (40%) spleen samples from the same cases. Positive results were not obtained with liver biopsy samples from yellow fever or Neisseria meningitides and Flu-A cases. 4H2 mAb recognizes the native protein of the four DENV serotypes in infected cells and did not cross-react with native ZIKV- or CHKV-infected cells by immunohistochemical assay, so it is a useful tool for differential histopathological conclusion of acute febrile hemorrhagic deaths.

Prevalence and factors associated with sepsis and septic shock in oncological patients in intensive therapy.

OBJECTIVES: to analyze factors associated with sepsis and septic shock in cancer patients in the Intensive Care Unit. METHODS: cross-sectional, retrospective study with a quantitative approach, with a sample of 239 patients in an oncology hospital. Secondary data from medical records were used. The outcome variable was "presence of sepsis and/or septic shock"; and exposures: sex, length of stay, origin, use of invasive procedures and primary tumor site. Descriptive, bivariate analyzes and multiple logistic regression models were performed. RESULTS: the prevalence of sepsis was 95% CI: 14.7-24.7 and septic shock of 95% CI: 37.7-50.3. In the multiple analysis, sepsis and/or septic shock were associated with hospital stay longer than seven days, being from the Emergency Department, presence of invasive procedures and hematological site. CONCLUSIONS: sepsis and/or septic shock in cancer patients were associated with clinical characteristics and health care factors.

The proposal of an international convention on response to pandemics: in defense of a human rights treaty for the field of global health / A proposta de convenção internacional sobre a resposta às pandemias: em defesa de um tratado de direitos humanos para o campo da saúde global

In November 2021, the World Health Organization (WHO) will start a negotiation of a convention, agreement, or other international instrument on response to pandemics. In this essay, we state and justify the thesis that the new pact shall be a human rights treaty as an imperative condition for the prevention of new pandemics and an efficient global response when they occur. After a brief description of the origin of the negotiations, we present thefollowing normative topics that correspond to a human rights perspective: the establishment of a rule of indissociability between quarantine measures and social protection; and the regulation of the access to pharmaceutical technologies. After that, in a section dedicated to the topics of the effectiveness of the future treaty, we classify the existing proposals in technocratic agreements, such as alterations in the procedures for the declaration of emergencies; mechanisms of transparency and accountability, such as the adoption of an Universal Periodic Reviews (UPR), similar to those of the United Nations Human Rights Council, to monitor the health-related obligations of States; coercive powers that would be disposed to WHO or to another agency such as inspections in the national territories to be held independent scientists; and mechanisms of political coordination, such as the creation of a Global Council on Threats to Health. We conclude that there is a risk of adopting a more efficient system to alert the developing world about threats originating from developing countries, in place of a treaty capable of contributing to avoid that more vulnerable populations continue to be devastated by even more frequent pandemics.

Em novembro de 2021, a Organização Mundial da Saúde (OMS) dará início à negociação de uma convenção, acordo ou outro instrumento internacional sobre a resposta às pandemias. Neste ensaio, defendemos e justificamos a tese de que o novo pacto deve ser um tratado de direitos humanos, como condição indispensável para prevenção de novas pandemias e eficiência da resposta global quando elas ocorrem. Após o breve resgate da origem das negociações, apresentamos os principais conteúdos normativos que correspondem a um enfoque de direitos humanos: a instituição da regra de indissociabilidade entre medidas quarentenárias e de proteção social; e a regulamentação do acesso a tecnologias farmacêuticas. A seguir, em seção dedicada ao tema da efetividade do futuro tratado, classificamos as propostas existentes em ajustes tecnocráticos, como alterações no procedimento de declaração de emergências; mecanismos de transparência e controle, a exemplo da adoção de um mecanismo de Revisão Periódica Universal (RPU), similar ao do Conselho de Direitos Humanos das Nações Unidas, para monitorar obrigações dos Estados relacionadas à saúde; poderes coercitivos que seriam outorgados à OMS ou outra agência, tais como inspeções nos territórios nacionais realizadas por cientistas independentes; e mecanismos de coordenação política, como a criação de um Conselho Global de Ameaças à Saúde. Concluímos que há risco de adoção de um sistema mais eficiente de vigilância para alertar o mundo desenvolvido sobre ameaças oriundas de países em desenvolvimento, em lugar de um tratado capaz de contribuir para evitar que populações mais vulneráveis continuem sendo devastadas por pandemias cada vez mais frequentes.

Human microbiota modulation via QseC sensor kinase mediated in the Escherichia coli O104:H4 outbreak strain infection in microbiome model.

BACKGROUND: The intestinal microbiota plays a crucial role in human health, adjusting its composition and the microbial metabolites protects the gut against invading microorganisms. Enteroaggregative E. coli (EAEC) is an important diarrheagenic pathogen, which may cause acute or persistent diarrhea (≥14 days). The outbreak strain has the potent Shiga toxin, forms a dense biofilm and communicate via QseBC two-component system regulating the expression of many important virulence factors. RESULTS: Herein, we investigated the QseC histidine sensor kinase role in the microbiota shift during O104:H4 C227-11 infection in the colonic model SHIME® (Simulator of the Human Intestinal Microbial Ecosystem) and in vivo mice model. The microbiota imbalance caused by C227-11 infection affected ỿ-Proteobacteria and Lactobacillus spp. predominance, with direct alteration in intestinal metabolites driven by microbiota change, such as Short-chain fatty acids (SCFA). However, in the absence of QseC sensor kinase, the microbiota recovery was delayed on day 3 p.i., with change in the intestinal production of SCFA, like an increase in acetate production. The higher predominance of Lactobacillus spp. in the microbiota and significant augmented qseC gene expression levels were also observed during C227-11 mice infection upon intestinal depletion. Novel insights during pathogenic bacteria infection with the intestinal microbiota were observed. The QseC kinase sensor seems to have a role in the microbiota shift during the infectious process by Shiga toxin-producing EAEC C227-11. CONCLUSIONS: The QseC role in C227-11 infection helps to unravel the intestine microbiota modulation and its metabolites during SHIME® and in vivo models, besides they contribute to elucidate bacterial intestinal pathogenesis and the microbiota relationships.