AB2CO risk score for in-hospital mortality of COVID-19 patients admitted to intensive care units.

PURPOSE: To develop a mortality risk score for COVID-19 patients admitted to intensive care units (ICU), and to compare it with other existing scores. MATERIALS AND METHODS: This retrospective observational study included consecutive adult patients with laboratory-confirmed COVID-19 admitted to ICUs of 18 hospitals from nine Brazilian cities, from September 2021 to July 2022. Potential predictors were selected based on the literature review. Generalized Additive Models were used to examine outcomes and predictors. LASSO regression was used to derive the mortality score. RESULTS: From 558 patients, median age was 69 years (IQR 58-78), 56.3 % were men, 19.7 % required mechanical ventilation (MV), and 44.8 % died. The final model comprised six variables: age, pO2/FiO2, respiratory function (respiratory rate or if in MV), chronic obstructive pulmonary disease, and obesity. The AB2CO had an AUROC of 0.781 (95 % CI 0.744 to 0.819), good overall performance (Brier score = 0.191) and an excellent calibration (slope = 1.063, intercept = 0.015, p-value = 0.834). The model was compared with other scores and displayed better discrimination ability than the majority of them. CONCLUSIONS: The AB2CO score is a fast and easy tool to be used upon ICU admission.

Clinical characteristics and outcomes of COVID-19 patients with preexisting dementia: a large multicenter propensity-matched Brazilian cohort study.

BACKGROUND: Although dementia has emerged as an important risk factor for severe SARS-CoV-2 infection, results on COVID-19-related complications and mortality are not consistent. We examined the clinical presentations and outcomes of COVID-19 in a multicentre cohort of in-hospital patients, comparing those with and without dementia. METHODS: This retrospective observational study comprises COVID-19 laboratory-confirmed patients aged ≥ 60 years admitted to 38 hospitals from 19 cities in Brazil. Data were obtained from electronic hospital records. A propensity score analysis was used to match patients with and without dementia (up to 3:1) according to age, sex, comorbidities, year, and hospital of admission. Our primary outcome was in-hospital mortality. We also assessed admission to the intensive care unit (ICU), invasive mechanical ventilation (IMV), kidney replacement therapy (KRT), sepsis, nosocomial infection, and thromboembolic events. RESULTS: Among 1,556 patients included in the study, 405 (4.5%) had a diagnosis of dementia and 1,151 were matched controls. When compared to matched controls, patients with dementia had a lower frequency of dyspnoea, cough, myalgia, headache, ageusia, and anosmia; and higher frequency of fever and delirium. They also had a lower frequency of ICU admission (32.7% vs. 47.1%, p < 0.001) and shorter ICU length of stay (7 vs. 9 days, p < 0.026), and a lower frequency of sepsis (17% vs. 24%, p = 0.005), KRT (6.4% vs. 13%, p < 0.001), and IVM (4.6% vs. 9.8%, p = 0.002). There were no differences in hospital mortality between groups. CONCLUSION: Clinical manifestations of COVID-19 differ between older inpatients with and without dementia. We observed that dementia alone could not explain the higher short-term mortality following severe COVID-19. Therefore, clinicians should consider other risk factors such as acute morbidity severity and baseline frailty when evaluating the prognosis of older adults with dementia hospitalised with COVID-19.

Recent advances in boron removal in aqueous media. An approach to the adsorption process and process optimization.

The numerous oxidation states of the element boron bring great challenges in containing its contamination in receptor bodies. This scenario increases significantly due to the widespread use of boron compounds in various industries in recent years. For this reason, the removal of this contaminant is receiving worldwide attention. Although adsorption is a promising method in boron removal, finding suitable adsorbents, that is, those with high efficiency, and feasible remains a constant challenge. Hence, this review presents the boron removal methods in comparison to costs of adsorbents, reaction mechanisms, economic viability, continuous bed application, and regeneration capacity. In addition, the approach of multivariate algorithms in the solution of multiobjective problems can enable the optimized conditions of dosage of adsorbents and coagulants, pH, and initial concentration of boron. Therefore, this review sought to comprehensively and critically demonstrate strategic issues that may guide the choice of method and adsorbent or coagulant material in future research for bench and industrial scale boron removal.

A novel vaccine based on SARS-CoV-2 CD4+ and CD8+ T cell conserved epitopes from variants Alpha to Omicron.

COVID-19 caused, as of September, 1rst, 2022, 599,825,400 confirmed cases, including 6,469,458 deaths. Currently used vaccines reduced severity and mortality but not virus transmission or reinfection by different strains. They are based on the Spike protein of the Wuhan reference virus, which although highly antigenic suffered many mutations in SARS-CoV-2 variants, escaping vaccine-generated immune responses. Multiepitope vaccines based on 100% conserved epitopes of multiple proteins of all SARS-CoV-2 variants, rather than a single highly mutating antigen, could offer more long-lasting protection. In this study, a multiepitope multivariant vaccine was designed using immunoinformatics and in silico approaches. It is composed of highly promiscuous and strong HLA binding CD4+ and CD8+ T cell epitopes of the S, M, N, E, ORF1ab, ORF 6 and ORF8 proteins. Based on the analysis of one genome per WHO clade, the epitopes were 100% conserved among the Wuhan-Hu1, Alpha, Beta, Gamma, Delta, Omicron, Mµ, Zeta, Lambda and R1 variants. An extended epitope-conservancy analysis performed using GISAID metadata of 3,630,666 SARS-CoV-2 genomes of these variants and the additional genomes of the Epsilon, Lota, Theta, Eta, Kappa and GH490 R clades, confirmed the high conservancy of the epitopes. All but one of the CD4 peptides showed a level of conservation greater than 97% among all genomes. All but one of the CD8 epitopes showed a level of conservation greater than 96% among all genomes, with the vast majority greater than 99%. A multiepitope and multivariant recombinant vaccine was designed and it was stable, mildly hydrophobic and non-toxic. The vaccine has good molecular docking with TLR4 and promoted, without adjuvant, strong B and Th1 memory immune responses and secretion of high levels of IL-2, IFN-γ, lower levels of IL-12, TGF-ß and IL-10, and no IL-6. Experimental in vivo studies should validate the vaccine's further use as preventive tool with cross-protective properties.

Coronavirus Disease 2019 Vaccination for Cancer Patients: Risk or Benefit?

OBJECTIVE: The aim of the present study is to list the published clinical trials on coronavirus disease 2019 (COVID-19) vaccines, to describe the mechanism of action of the identified vaccines, and to identify protocols regarding safety, status, and prioritization of cancer patients for vaccination. METHODS: This is a systematic review with a limited literature search conducted by an information specialist; key resources such as PubMed and websites of major cancer organizations were searched. The main search terms were COVID-19, vaccination, cancer, and breast and gynecological cancers. RESULTS: Cancer patients infected with the new coronavirus are at high risk of complications and death, but we still know little about the risks and benefits of vaccination for COVID-19 in these patients. In an ideal scenario, all cancer patients should have their immunization status updated before beginning treatment, but this is not always possible. CONCLUSION: Patients with breast or gynecological cancers who are receiving treatment or are in the 5-year posttreatment period should be included in the priority group for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) vaccination.

OBJETIVO: O objetivo do presente estudo é listar os ensaios clínicos publicados sobre as vacinas para coronavirus disease 2019 (COVID-19), descrever seus mecanismos de ação e descrever protocolos sobre segurança, status e priorização de pacientes oncológicos para vacinação. MéTODOS: Trata-se de uma revisão sistemática com uma pesquisa bibliográfica limitada conduzida por um especialista em informação; bases de dados como PubMed e sites das principais organizações de câncer foram pesquisados. Os principais termos de pesquisa foram COVID-19, vacinação, câncer e câncer de mama e ginecológico. RESULTADOS: Pacientes com câncer infectados com o novo coronavírus têm alto risco de complicações e morte, mas ainda sabemos pouco sobre os riscos e benefícios da vacinação para COVID-19 nesses pacientes. Em um cenário ideal, todos os pacientes com câncer deveriam ter seu estado de imunização atualizado antes de iniciar o tratamento, mas nem sempre isso é possível. CONCLUSãO: Pacientes com câncer de mama ou ginecológico em tratamento ou no período pós-tratamento de 5 anos devem ser incluídos no grupo prioritário para vacinação contra severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2.).

Coronavirus Disease 2019 Vaccination for Cancer Patients: Risk or Benefit? / Vacinação para COVID-19 em pacientes oncológicos: Risco ou benefício?

Abstract Objective The aim of the present study is to list the published clinical trials on coronavirus disease 2019 (COVID-19) vaccines, to describe the mechanism of action of the identified vaccines, and to identify protocols regarding safety, status, and prioritization of cancer patients for vaccination. Methods This is a systematic review with a limited literature search conducted by an information specialist; key resources such as PubMed and websites of major cancer organizations were searched. The main search terms were COVID-19, vaccination, cancer, and breast and gynecological cancers. Results Cancer patients infected with the new coronavirus are at high risk of complications and death, but we still know little about the risks and benefits of vaccination for COVID-19 in these patients. In an ideal scenario, all cancer patients should have their immunization status updated before beginning treatment, but this is not always possible. Conclusion Patients with breast or gynecological cancers who are receiving treatment or are in the 5-year posttreatment period should be included in the priority group for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) vaccination.

Resumo Objetivo O objetivo do presente estudo é listar os ensaios clínicos publicados sobre as vacinas para coronavirus disease 2019 (COVID-19), descrever seus mecanismos de ação e descrever protocolos sobre segurança, status e priorização de pacientes oncológicos para vacinação. Métodos Trata-se de uma revisão sistemática com uma pesquisa bibliográfica limitada conduzida por um especialista em informação; bases de dados como PubMed e sites das principais organizações de câncer foram pesquisados. Os principais termos de pesquisa foram COVID-19, vacinação, câncer e câncer de mama e ginecológico. Resultados Pacientes com câncer infectados com o novo coronavírus têm alto risco de complicações e morte, mas ainda sabemos pouco sobre os riscos e benefícios da vacinação para COVID-19 nesses pacientes. Em um cenário ideal, todos os pacientes com câncer deveriam ter seu estado de imunização atualizado antes de iniciar o tratamento, mas nem sempre isso é possível. Conclusão Pacientes com câncer de mama ou ginecológico em tratamento ou no período pós-tratamento de 5 anos devem ser incluídos no grupo prioritário para vacinação contra severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2.)

Cardiometabolic Effects of Postnatal High-Fat Diet Consumption in Offspring Exposed to Maternal Protein Restriction In Utero.

In recent decades, the high incidence of infectious and parasitic diseases has been replaced by a high prevalence of chronic and degenerative diseases. Concomitantly, there have been profound changes in the behavior and eating habits of families around the world, characterizing a "nutritional transition" phenomenon, which refers to a shift in diet in response to modernization, urbanization, or economic development from undernutrition to the excessive consumption of hypercaloric and ultra-processed foods. Protein malnutrition that was a health problem in the first half of the 20th century has now been replaced by high-fat diets, especially diets high in saturated fat, predisposing consumers to overweight and obesity. This panorama points us to the alarming coexistence of both malnutrition and obesity in the same population. In this way, individuals whose mothers were undernourished early in pregnancy and then exposed to postnatal hyperlipidic nutrition have increased risk factors for developing metabolic dysfunction and cardiovascular diseases in adulthood. Thus, our major aim was to review the cardiometabolic effects resulting from postnatal hyperlipidic diets in protein-restricted subjects, as well as to examine the epigenetic repercussions occasioned by the nutritional transition.

Triple-negative breast cancer treatment in xenograft models by bifunctional nanoprobes combined to photodynamic therapy.

Triple-negative breast cancer (TNBC) overexpresses the Epidermal Growth Factor Receptor (EGFR), a characteristic of different types of tumors, linked to worse disease prognosis and risk of recurrence. Conventional treatments are also aggressive and can be morbid.. Therefore, t improvement and development of new methods are notorious. Photodynamic Therapy (PDT) is an effective method for treating different types of cancer by using light radiation to activate a photosensitizing agent (drug) in molecular oxygen presence, promoting cell death., Improving drug uptake in target cells could contribute to PDT efficiency. Accordingly, we developed a bifunctional nanoprobe (BN), used in PDT as a a treatment method in vivo against breast cancer. The BN uses gold nanoparticles with active targeting through the Epidermal Growth Factor (EGF) protein and Chlorine e6 (Ce6) carriers. We evaluated the therapeutic efficacy of in vivo xenograft in 4 groups: Saline, BN, Ce6+PDT, and BN+PDT. As a result, we observed that the BN+PDT group exhibited an excellent effect with greater selectivity to tumor tissue and tissue damage when compared to the Saline, BN, and Ce6+PDT groups. The results indicate a potential impact on breast cancer treatment in vivo.. In conclusion, our data propose that the BN developed heightened PDT efficacy through cellular DNA repair effects and tumor microenvironment.

Avaliação da expressão da filagrina em biópsias esofágicas de pacientes com esofagite eosinofílica / Evaluation of philagrin expression in esophageal biopsies of patients with eosinophilic esophagitis

Introdução: Mutações do gene da filagrina vêm sendo associadas, classicamente, a alterações da barreira epitelial em doenças alérgicas com comprometimento da pele e das superfícies mucosas. Particularmente na dermatite atópica, a relação entre filagrina, mecanismo fisiopatológico e evolução clínica tem sido demonstrada. Recentemente, alterações da barreira epitelial com redução da expressão da filagrina, também têm sido associadas a mecanismos imunológicos envolvidos na patogênese da esofagite eosinofílica. Devido a disfunções na barreira epitelial, microrganismos e alérgenos são capazes de penetrarem no epitélio da mucosa esofágica, assim como na dermatite atópica. Objetivo: Avaliar a possível correlação da expressão da filagrina com os achados histopatológicos em biópsias esofágicas de pacientes com esofagite eosinofílica. Métodos: A expressão da filagrina foi investigada in situ, por imuno-histoquímica, em biópsias esofágicas nos seguintes grupos: Grupo I, controle (n=8), amostras provenientes de pacientes saudáveis; Grupo II (n=27), amostras provenientes de pacientes com esofagite eosinofílica. Resultados: Os resultados demonstraram uma diminuição da expressão da filagrina na mucosa do esôfago de portadores de esofagite eosinofílica. Adicionalmente, a intensidade da marcação imuno-histoquímica foi menor na mucosa esofágica com maior infiltração de eosinófilos. Conclusão: A diminuição da expressão de filagrina pode ser um fenomeno fisiopatológico associado ao aumento da quantidade de eosinófilos na mucosa esofágica, podendo impactar na evolução clínica da esofagite eosinofílica.

Introduction:Filaggrin gene mutations have been classically associated with changes in the epithelial barrier in allergic diseases involving the skin and mucosal surfaces. Particularly in atopic dermatitis, the relationship between filaggrin, pathophysiological mechanism and clinical evolution hás been demonstrated. Recently, changes in the epithelial barrier with reduced expression of filaggrin have also been associated with immunological mechanisms involved in the pathogenesis of eosinophilic esophagitis. Due to dysfunction in the epithelial barrier, microorganisms and allergens are able to penetrate the epithelium of the esophageal mucosa, as well as in atopic dermatitis. Objective: To evaluated the possible correlation of filaggrin expression with histopathological findings in esophageal biopsies of patients with eosinophilic esophagitis. Methods: Filaggrin expression was investigated in situ by immunohistochemistry in esophageal biopsies in the following groups: Group I, control (n = 8), samples from healthy patients; Group II (n = 27), samples from patients with eosinophilic esophagitis. Results: The results demonstrated a decrease in the expression of filaggrin in the esophageal mucosa of patients with eosinophilic esophagitis. Additionally, the intensity of the immunohistochemical labeling was lower in the esophageal mucosa with greater infiltration of eosinophils. Conclusion: The reduction of filaggrin expression may be a pathophysiological phenomenon associated with an increase in the quantity of eosinophils in the esophageal mucosa, which may impact on the clinical evolution of eosinophilic esophagitis.

Expressões do desamparo na escola: vulnerabilidades sociais e impactos na adolescência / Expressions of helplessness at school: social vulnerabilities and impacts on adolescence

O artigo deriva de uma pesquisa em andamento que visa compreender como as condições de vulnerabilidade social em que se encontram muitos jovens brasileiros se articulam ao desamparo como efeito psíquico e, mais especificamente, de que modo a relação com a escola toma parte nos impasses da adolescência no laço social. Na interface entre a psicanálise e a educação, trabalharemos com o material levantado a partir da realização de oficinas com turmas do 9º ano do ensino fundamental ao 3º ano do ensino médio em duas escolas da rede pública do Estado do Rio de Janeiro. Os adolescentes se utilizaram do espaço da oficina para expressar seu desamparo, seja pelo silêncio, pelo afrontamento ou pela sua expressão em palavras nos segredos compartilhados nas oficinas, que foram agrupados do seguinte modo: desamparo em relação às condições materiais da escola; desamparo por não se sentir acolhido pela escola; desamparo na relação com seus pares; expressões do desamparo como desalento.

The article derives from an ongoing research that aims to understand how the conditions of social vulnerability in which many young Brazilians find themselves are articulated to helplessness as a psychic effect and, more specifically, how the relationship with the school takes part in the impasses of adolescence in the social bond. At the interface between psychoanalysis and education, we will work with the material raised from the realization of workshops with classes from the 9th grade of elementary school to the 3rd year of high school in two public schools in the state of Rio de Janeiro. The adolescents used the space of the workshop to express their helplessness, either by silence, by confrontation or by their expression in words in the secrets shared in the workshops, which were grouped as follows: helplessness in relation to the material conditions of the school; helplessness for not feeling welcomed by the school; helplessness in the relationship with their peers; expressions of helplessness as discouragement.

Factors driving extensive spatial and temporal fluctuations in COVID-19 fatality rates in Brazilian hospitals

The SARS-CoV-2 Gamma variant spread rapidly across Brazil, causing substantial infection and death waves. We use individual-level patient records following hospitalisation with suspected or confirmed COVID-19 to document the extensive shocks in hospital fatality rates that followed Gammas spread across 14 state capitals, and in which more than half of hospitalised patients died over sustained time periods. We show that extensive fluctuations in COVID-19 in-hospital fatality rates also existed prior to Gammas detection, and were largely transient after Gammas detection, subsiding with hospital demand. Using a Bayesian fatality rate model, we find that the geographic and temporal fluctuations in Brazils COVID-19 in-hospital fatality rates are primarily associated with geographic inequities and shortages in healthcare capacity. We project that approximately half of Brazils COVID-19 deaths in hospitals could have been avoided without pre-pandemic geographic inequities and without pandemic healthcare pressure. Our results suggest that investments in healthcare resources, healthcare optimization, and pandemic preparedness are critical to minimize population wide mortality and morbidity caused by highly transmissible and deadly pathogens such as SARS-CoV-2, especially in low- and middle-income countries. NoteThe following manuscript has appeared as Report 46 - Factors driving extensive spatial and temporal fluctuations in COVID-19 fatality rates in Brazilian hospitals at https://spiral.imperial.ac.uk:8443/handle/10044/1/91875. One sentence summaryCOVID-19 in-hospital fatality rates fluctuate dramatically in Brazil, and these fluctuations are primarily associated with geographic inequities and shortages in healthcare capacity.

Avaliação do emprego da imunoglobulina humana endovenosa sobre a densidade de infecções em pacientes com mieloma múltiplo / Evaluation of the use of human intravenous immunoglobulin on the incidence density of infections in patients with multiple myeloma

Introdução: O mieloma múltiplo (MM) é uma neoplasia hematológica que cursa com hipogamaglobulinemia e consequente imunodeficiência secundária. Uma das principais causas de morbimortalidade desses pacientes são infecções. Objetivou-se com esse estudo avaliar o impacto da reposição de imunoglobulina endovenosa (IgIV) na taxa de infecções em pacientes portadores de MM. Métodos: Trata-se de um estudo de análise documental, com variáveis qualitativas e quantitativas, com objetivo de realizar análise retrospectiva dos prontuários de pacientes com MM que receberam tratamento com imunoglobulina humana endovenosa em um hospital privado na cidade de Patos de Minas, MG, Brasil, no período de 01/05/2016 a 31/12/2020. Foram coletados dados epidemiológicos, resultados de exames, episódios de infecções, eventos adversos da medicação e desfecho dos pacientes nos prontuários analisados. Resultados: Foram identificados 10 pacientes com diagnóstico de MM, todos receberam IgIV na dose de 300 a 400 mg/kg/mês. Nenhuma reação adversa relacionada ao uso da IgIV foi registrada nos prontuários. Foram identificados seis quadros infecciosos que ocorreram em quatro pacientes. Nenhum diagnóstico de sepse foi registrado. A densidade de incidência de infecções foi de 0,28 episódios/pacientes-ano. Conclusão: A densidade de incidência de infecções observada no presente estudo foi significativamente menor em comparação ao que se tem registro na literatura, sugerindo importante papel da IgIV na prevenção de infecções em pacientes com MM.

Introduction: Multiple myeloma (MM) is a hematologic malignancy that leads to hypogammaglobulinemia and consequent secondary immunodeficiency. Infections are a major cause of morbidity and mortality in these patients. The objective of this study was to evaluate the impact of intravenous immunoglobulin (IVIg) replacement on the rate of infections in patients with MM. Methods: This document analysis study used qualitative and quantitative variables to perform a retrospective analysis of the medical records of patients with MM who were treated with human IVIg in a private hospital in the city of Patos de Minas, MG, Brazil, from May 1, 2016 to December 31, 2020. Epidemiological data, test results, episodes of infections, adverse medication events, and patient outcomes were collected from the medical records. Results: Ten patients diagnosed with MM were identified, and they all received IVIg at a dose of 300 to 400 mg/kg/month. No adverse reactions related to the use of IVIg were recorded. Six infections that occurred in 4 patients were identified. No diagnosis of sepsis was recorded. The incidence density of infections was 0.28 episodes/patient-years. Conclusion: The incidence density of infections was significantly smaller in this study in comparison with previous literature findings, which suggests a significant role of IVIg in the prevention of infections in patients with MM.

Novel phage display-derived recombinant antibodies recognizing both MPT64 native and mutant (63-bp deletion) are promising tools for tuberculosis diagnosis.

Tuberculosis (TB) is one of the top 10 causes of death in humans worldwide. The most important causative agents of TB are bacteria from the Mycobacterium tuberculosis complex (MTC), although nontuberculous mycobacteria (NTM) can also cause similar infections. The ability to identify and differentiate MTC isolates from NTM is important for the selection of the correct antimicrobial therapy. Immunochromatographic assays with antibodies anti-MPT64 allow differentiation between MTC and NTM since the MPT64 protein is specific from MTC. However, studies reported false-negative results mainly due to mpt64 63-bp deletion. Considering this drawback, we selected seven human antibody fragments against MPT64 by phage display and produced them as scFv-Fc. Three antibodies reacted with rMPT64 mutant (63-bp deletion) protein and native MPT64 from M. tuberculosis H37Rv in ELISA and Western blot. These antibodies are new biological tools with the potential for the development of TB diagnosis helping to overcome limitations of the MPT64-based immunochromatographic tests currently available.

Satisfaction of allergic patients treated with house dust mite sublingual immunotherapy.

Sublingual immunotherapy (SLIT) has been used for more than three decades as a therapeutic strategy for the treatment of allergic diseases. Studies have demonstrated its efficacy and safety, and numerous clinical trials have evaluated these parameters. In the present study, through patient perception, we investigated the patient satisfaction with the use of house dust mite SLIT treatment. "Satisfaction Scale for Patients Receiving Allergen Immunotherapy" (ESPIA) questionnaire, a standardized and validated instrument for clinical studies evaluating allergen immunotherapy, was applied to allergic patients (N = 136). Children and adults of both sexes who received SLIT for Dermatophagoides pteronyssinus and/or Blomia tropicalis, according to the results of an immediate reading puncture test, were included. Data analysis showed that the perception of treatment effectiveness was 92%, performance improvement in the daily activities was 91%, a satisfactory cost-benefit balance was 84%, and the perception of general satisfaction was 97%. The results showed a high perception of satisfaction in allergic patients undergoing house dust mite SLIT.

Novel phage display-derived recombinant antibodies recognizing both MPT64 native and mutant (63-bp deletion) are promising tools for tuberculosis diagnosis

Tuberculosis (TB) is one of the top 10 causes of death in humans worldwide. The most important causative agents of TB are bacteria from the Mycobacterium tuberculosis complex (MTC), although nontuberculous mycobacteria (NTM) can also cause similar infections. The ability to identify and differentiate MTC isolates from NTM is important for the selection of the correct antimicrobial therapy. Immunochromatographic assays with antibodies anti-MPT64 allow differentiation between MTC and NTM since the MPT64 protein is specific from MTC. However, studies reported false-negative results mainly due to mpt64 63-bp deletion. Considering this drawback, we selected seven human antibody fragments against MPT64 by phage display and produced them as scFv-Fc. Three antibodies reacted with rMPT64 mutant (63-bp deletion) protein and native MPT64 from M. tuberculosis H37Rv in ELISA and Western blot. These antibodies are new biological tools with the potential for the development of TB diagnosis helping to overcome limitations of the MPT64-based immunochromatographic tests currently available.

Mapping the Brazilian microscopy landscape: A bibliometric and network analysis.

Understanding how a technology is introduced and shared in a society has a strategic value for the planning of technological development and assessing new market opportunities. Among other technologies, microscopy has had a significant role in advancing different fields of science. In Brazil, its use spans from biomedical to engineering areas. Here, we used social network analysis (SNA) to map and quantify the flow of interaction between Brazilian researchers involved in microscopy techniques. The analysis examines co-occurrence of thematic networks and scientific co-authorship in articles published in a ten years window, as retrieved from Scopus database. The results showed an increasing volume of publications using microscopy in Brazil. The two major areas of interest are material and life sciences, which present significant intra-regional interaction. USA, Spain, Germany, Portugal and the United Kingdom are the main partner countries for international scientific collaborations. The share of Brazilian publications applying microscopy follows the global trends, with a slight predominance in health and life sciences. Our results provide a context of the strengths and gaps of the field in Brazil and may help to inform researchers and policy makers for further advancing the field.

Protection efficacy of the rLTB-R1 chimera against experimental swine mycoplasmal pneumonia

Background: Mycoplasma hyopneumoniae is the etiological agent of the Swine Mycoplasmal Pneumonia (SMP), one of the most economically significant diseases in the swine industry worldwide. Commonly used vaccines for SMP control consist of inactivated whole cells (bacterins). These vaccines are efficacious against M. hyopneumoniae challenge, but do not prevent colonization by the pathogen or completely eliminate pneumonia. P97 adhesin is conserved in the M. pneumoniae virulent strains, therefore it is an attractive target to be used in recombinant vaccines against M. hyopneumoniae. The aim of the present study was to evaluate protection afforded by rLTB-R1, a recombinant chimera composed by LTB fused with the R1 repeat region of P97 adhesin of M. hyopneumoniae, in specific-pathogen-free (SPF) piglets vaccinated by intranasal or intramuscular route and challenged with a pathogenic strain of M. hyopneumoniae. Materials, Methods & Results: PCR products of the LTB and R1 coding sequences were fused, then cloned into pETDEST42™ expression vector. The rLTB-R1 was expressed in Escherichia coli BL21 (DE3) Salt induction (SI). The piglets were divided into three groups: four piglets were intranasally vaccinated with 1 mg of rLTB-R1 solubilized in 1 mL of PBS at 0 and 14 days (IN rLTB-R1 group); four piglets were intramuscularly vaccinated with 1 mg of rLTB-R1 solubilized in 1 mL of PBS at 0 and 14 days (IM rLTB-R1 group); three piglets were intranasally and intramuscularly inoculated with 1 mL of PBS (control group). Two weeks after the last immunization (28 day), piglets were intratracheally challenged with 10 mL of a suspension containing 109 color-changing unit (CCU) of pathogenic M. hyopneumoniae 7448 strain on three consecutive days. Until the challenge (28 days), intranasal and intramuscular vaccination with rLTB-R1 induced seroconversions of antiR1 systemic antibodies of 1.6 and 4.6 ×, respectively. The IN rLTB-R1 group had no pulmonary lesion, rLTB-R1 conferred protection against experimental SMP. On the other hand, IM rLTB-R1 and control groups had on average 7.24% and 8.46% of pulmonary lesion, respectively, showing that intramuscular vaccination with rLTB-R1 did not confer protection. Discussion: The rLTB-R1, when intranasally administrated to mice, elicited production of anti-R1 IgA in trachea and bronchi as well as specific Th1 response, suggesting an adequate stimulation of the mucosal immune system. We believe that rLTB-R1 induced a similar immune response in piglets intranasally vaccinated, conferring protection against experimental SMP. The present study, the rLTB-R1 alone, without any chemical adjuvant, stimulated a significant seroconversion of anti-R1 systemic antibodies in pigs intramuscularly vaccinated, showing the potential of LTB as a parenteral adjuvant in swine vaccination. Previous work has shown that the intramuscular administration route was evaluated in pigs because mice intramuscularly vaccinated with rLTB-R1 presented significant levels of anti-R1 IgA in trachea and bronchi, suggesting that rLTB can stimulate some degree of mucosal immunity even if not delivered by a mucosal route. However, in the present study, piglets intramuscularly vaccinated with rLTB-R1 presented high levels of anti-R1 systemic antibodies, they were not protected. On the other hand, intranasal vaccination of piglets with rLTB-R1 elicited low levels of antiR1 systemic antibodies (1.6 × at 28 days), but it conferred full protection against experimental SMP. The present study demonstrated that intranasal vaccination of piglets with rLTB-R1 conferred protection against experimental SMP. A more detailed analysis of the protective immune response induced by rLTB-R1 in pigs is currently being performed.

Protection efficacy of the rLTB-R1 chimera against experimental swine mycoplasmal pneumonia

Background: Mycoplasma hyopneumoniae is the etiological agent of the Swine Mycoplasmal Pneumonia (SMP), one of the most economically significant diseases in the swine industry worldwide. Commonly used vaccines for SMP control consist of inactivated whole cells (bacterins). These vaccines are efficacious against M. hyopneumoniae challenge, but do not prevent colonization by the pathogen or completely eliminate pneumonia. P97 adhesin is conserved in the M. pneumoniae virulent strains, therefore it is an attractive target to be used in recombinant vaccines against M. hyopneumoniae. The aim of the present study was to evaluate protection afforded by rLTB-R1, a recombinant chimera composed by LTB fused with the R1 repeat region of P97 adhesin of M. hyopneumoniae, in specific-pathogen-free (SPF) piglets vaccinated by intranasal or intramuscular route and challenged with a pathogenic strain of M. hyopneumoniae. Materials, Methods & Results: PCR products of the LTB and R1 coding sequences were fused, then cloned into pETDEST42™ expression vector. The rLTB-R1 was expressed in Escherichia coli BL21 (DE3) Salt induction (SI). The piglets were divided into three groups: four piglets were intranasally vaccinated with 1 mg of rLTB-R1 solubilized in 1 mL of PBS at 0 and 14 days (IN rLTB-R1 group); four piglets were intramuscularly vaccinated with 1 mg of rLTB-R1 solubilized in 1 mL of PBS at 0 and 14 days (IM rLTB-R1 group); three piglets were intranasally and intramuscularly inoculated with 1 mL of PBS (control group). Two weeks after the last immunization (28 day), piglets were intratracheally challenged with 10 mL of a suspension containing 109 color-changing unit (CCU) of pathogenic M. hyopneumoniae 7448 strain on three consecutive days. Until the challenge (28 days), intranasal and intramuscular vaccination with rLTB-R1 induced seroconversions of antiR1 systemic antibodies of 1.6 and 4.6 ×, respectively. The IN rLTB-R1 group had no pulmonary lesion, rLTB-R1 conferred protection against experimental SMP. On the other hand, IM rLTB-R1 and control groups had on average 7.24% and 8.46% of pulmonary lesion, respectively, showing that intramuscular vaccination with rLTB-R1 did not confer protection. Discussion: The rLTB-R1, when intranasally administrated to mice, elicited production of anti-R1 IgA in trachea and bronchi as well as specific Th1 response, suggesting an adequate stimulation of the mucosal immune system. We believe that rLTB-R1 induced a similar immune response in piglets intranasally vaccinated, conferring protection against experimental SMP. The present study, the rLTB-R1 alone, without any chemical adjuvant, stimulated a significant seroconversion of anti-R1 systemic antibodies in pigs intramuscularly vaccinated, showing the potential of LTB as a parenteral adjuvant in swine vaccination. Previous work has shown that the intramuscular administration route was evaluated in pigs because mice intramuscularly vaccinated with rLTB-R1 presented significant levels of anti-R1 IgA in trachea and bronchi, suggesting that rLTB can stimulate some degree of mucosal immunity even if not delivered by a mucosal route. However, in the present study, piglets intramuscularly vaccinated with rLTB-R1 presented high levels of anti-R1 systemic antibodies, they were not protected. On the other hand, intranasal vaccination of piglets with rLTB-R1 elicited low levels of antiR1 systemic antibodies (1.6 × at 28 days), but it conferred full protection against experimental SMP. The present study demonstrated that intranasal vaccination of piglets with rLTB-R1 conferred protection against experimental SMP. A more detailed analysis of the protective immune response induced by rLTB-R1 in pigs is currently being performed.

Combined use of ELISA and Western blot with recombinant N protein is a powerful tool for the immunodiagnosis of avian infectious bronchitis.

BACKGROUND: The avian infectious bronchitis virus (IBV) remains a significant source of loss in the poultry industry and early diagnosis is required to prevent the disease from spreading. This study examined the combined use of an ELISA and Western blot (WB) to detect antibodies against the nucleocapsid protein (N) of IBV. The coding sequence for N was amplified by RT-PCR and expressed in Escherichia coli. A soluble recombinant N protein (rN) of approximately 50 kDa was obtained. A total of 389 sera were tested against the rN in ELISA and the results were compared with those of the commercial IDEXX IBV Ab test. ELISA-rN achieved a 90.34% sensitivity and 90.16% specificity. WB confirmed all false negative sera in ELISA-rN or IDEXX test as truly positive. The current study indicate that the combined use of rN in ELISA and WB is a powerful tool for the immunodiagnosis of avian infectious bronchitis. METHODS: Constructed recombinant pAE/n expression vectors were used to transform E. coli BL21(DE3) Star competent cells (Invitrogen). The rN of infectious bronchitis virus was purified by affinity chromatography using HisTrap HP 1 mL columns pre-packed with pre-charged Ni Sepharose in the ÄKTAprime Automated Liquid Chromatography system (GE Healthcare). A total of 389 serum samples from chickens were used to develop and evaluate the ELISA-rN test. To standardize the indirect ELISA development, serum dilutions (1:100, 1:200 and 1:400) and different concentrations of purified rN antigen (50, 100 and 200 ng/well) were tested. Positive and negative sera for IBV were used as controls. The results were compared with those obtained from a commercial kit. Serum samples scored as negative with the commercial kit but as positive with the ELISA-rN were further analysed by Western blot analyses using the rN protein as an antigen. The results of the ELISA-rN were compared to the commercial kit results using receiver-operating characteristics curves, area under the curve, and confidence intervals with the software GraphPad Prism version 6.0 for Windows (GraphPad Software, USA). RESULTS: The expected cDNA fragment of approximately 1240 bp was successfully amplified by PCR using primers designed to select for the coding region of the N protein. The rN was expressed as a soluble protein to avoid the refolding steps and, after purification a yield of 10 mg/L of rN was obtained. The SDS-PAGE results demonstrated the presence of two distinct bands that had a molecular mass of approximately 45 and 50 KDa. Out of 244 sera that scored positive in the commercial ELISA IDEXX IBV Ab Test, 220 were also positive in the ELISA-rN, yielding an ELISA-rN test sensitivity of 90.16%. Out of 145 sera that scored negative in the IDEXX IBV Ab Test, 131 also scored negative in the ELISA-rN, indicating a specificity of 90.34%. Sera that tested negative in the ELISA-rN and positive in the commercial test also reacted with the rN protein in Western blot. CONCLUSIONS: The association between the ELISA and Western blot techniques developed in this study with a subunit of IBV (rN) were able to detect antibodies that the commercial ELISA did not detect suggesting that the ELISA-rN has greater sensitivity.

Assessing mercury intoxication in isolated/remote populations: Increased S100B mRNA in blood in exposed riverine inhabitants of the Amazon.

Mercury is a heavy metal responsible for human intoxication worldwide and especially in the Amazon, where both natural and anthropogenic sources are responsible for exposure in riverine populations. Methylmercury is the most toxic specie of mercury with recognized neurotoxicity due to its affinity for the central nervous system. S100B protein is a well-established biomarker of brain damage and it was recently associated with mercury-related neurotoxicity. Accurate measurement is especially challenging in isolated/remote populations due to the difficulty of adequate sample conservation, therefore here we use S100B mRNA levels in blood as a way to assay mercury neurotoxicity. We hypothesized that individuals from chronically exposed populations showing mercury levels above the limit of 10 µg/g in hair would present increased levels of S100B mRNA, likely due to early brain damage. A total of 224 riverine individuals were evaluated for anthropometric data (age, body mass index), self-reported symptoms of mercury intoxication, c-reactive protein in blood, and mercury speciation in hair. Approximately 20% of participants showed mercury levels above the limit, and prevalence for most symptoms was not different between individuals exposed to high or low mercury levels. Rigorous exclusion criteria were applied to avoid confounding factors and S100B mRNA in blood was tested by RT-qPCR. Participants with ≥10 µg/g of mercury had S100B mRNA levels over two times higher than that of individuals with lower exposure. A significant correlation was also detected between mercury content in hair and S100B mRNA levels in blood, supporting the use of the latter as a possible candidate to predict mercury-induced neurotoxicity. This is the first report of an association between S100B mRNA and mercury exposure in humans. The combination of both exposure and intoxication biomarkers could provide additional support for the screening and early identification of high-risk individuals in isolated populations and subsequent referral to specialized centers.