Expressão e análise antigênica da proteína RTP36 recombinante da amostra São Paulo de Ehrlichia canis para testes sorológicos

Ehrlichia canis is the main etiological agent of canine monocytic ehrlichiosis (CME), a globally canine infectious disease. In Brazil, CME is considered to be endemic, and its prevalence can reach 65% in some states. The diagnosis of ehrlichiosis is important for treatment and epidemiological purposes. The E. canis TRP36 (Tandem Repeat Protein) protein elicits the earliest acute-phase antibody response observed during the course of the disease. This study aimed to generate the recombinant TRP36 protein from E. canis São Paulo strain and to evaluate its potential as a tool for the serologic diagnosis of CME. The E. canis São Paulo isolate was cultivated in DH82 lineage cells, and its genomic DNA was obtained. The bacterial DNA fragment encoding the entire ORF of TRP36 was cloned into the pBAD/Thio-TOPO vector and transformed into Escherichia coli DH10B competent cells with the trp36-bearing plasmid for protein expression. To evaluate the protein antigenicity, 16 canine serum samples were previously tested (by PCR and the commercial SNAP4Dx serological test). The results were in accordance with the SNAP4Dx test. Experiments using this recombinant protein as an antigen, targeting the development of a serologic test based on ELISA methodology, are the next step to produce a reliable, affordable and useful diagnostic tool for CME in Brazil.Ehrlichia canis é o principal agente etiológico da erliquiose monocítica canina (EMC), uma doença infecciosa canina globalmente dispersa. No Brasil, a EMC é considerada endêmica, e a infecção pode atingir 65% em cães em alguns estados. O diagnóstico de erliquiose é importante para fins de tratamento e epidemiológicos. A proteína TRP36 de E. canis leva a uma resposta humoral com produção de anticorpos em fase aguda, encontrada durante o curso da doença. O objetivo deste estudo foi obter a proteína TRP36 recombinante da amostra São Paulo de E. canis e avaliar seu potencial como ferramenta para o diagnóstico sorológico da CME. O isolado de E. canis São Paulo foi cultivado em células da linhagem DH82 e o DNA genômico foi obtido. O fragmento de DNA bacteriano que codifica toda a ORF de TRP36 foi clonado no vetor pBAD / Thio-TOPO e transformado em células competentes Escherichia coli DH10B, com o plasmídeo portador de trp36 para expressão de proteínas. Para avaliar a antigenicidade da proteína, 16 amostras de soro canino foram previamente analisadas (por PCR e teste sorológico comercial SNAP4Dx). Os resultados estavam de acordo com o teste SNAP4Dx. Os experimentos que utilizam essa proteína recombinante como antígeno, visando ao desenvolvimento de um teste sorológico baseado no ELISA, são o próximo passo para produzir um teste de diagnóstico confiável, acessível e útil para o diagnóstico da EMC no Brasil

Ecobiology of Haemagogus leucocelaenus arbovirus vector in the golden lion tamarin translocation area of Rio de Janeiro, Brazil

Abstract Significant pathogens that have resurfaced in humans originate from transmission from animal to human populations. In the Americas, yellow fever cases in humans are usually associated with spillover from non-human primates via mosquitoes. The present study characterized the prevalence of the yellow fever vector Haemagogus leucocelaenus in Rio de Janeiro, Brazil. The Atlantic Forest fragment chosen is an area of translocation of the golden lion tamarin (Leontopithecus rosalia), where 10 ovitraps were installed to collect mosquito eggs in Fazenda Três Irmãos, at Silva Jardim city, from March 2020 to October 2022. A total of 1514 eggs were collected, of which 1153 were viable; 50% belonged to medically important mosquito species and 24% to the yellow fever vector species, Hg. leucocelaenus. The months of December 2020 (n = 252), November 2021 (n = 188), and January 2022 (n = 252) had the highest densities of this vector. Haemagogus leucocelaenus was positively correlated with temperature (r = 0.303) and humidity (r = 0.48), with eggs hatching up to the 15th immersion with higher abundance of females. Implementing mosquito monitoring for arbovirus activity can help protect both the golden lion tamarin and human populations from the threat of arbovirus transmission

Interaction of <em>Haemagogus leucocelaenus</em> (Diptera: Culicidae) and Other Mosquito Vectors in a Forested Area, Rio de Janeiro, Brazil

The yellow fever (YF) virus has been detected throughout Brazil, with the occurrence of human cases, cyclic epizootics, and its isolation from Haemagogus janthinomys and Hg. leucocelaenus. We assessed the seasonal occurrence, egg abundancy, and oviposition interaction of mosquito vector species captured at a Private Natural Heritage Reserve in the Atlantic Forest biome. A total of 2943 eggs and 1538 mosquito larvae were collected from which 1231 belonged to entomologically important species. Ovitraps were used to collect immature mosquitoes from September 2019 to January 2021. The Mann–Whitney test was used to assess the differences in the abundance of eggs between rainy and dry seasons. Kruskal–Wallis and Dunn’s post hoc tests were used to evaluate the significance of the differences in the number of individuals from vector species. The highest percentage of mosquito vector eggs were collected during the rainy season, from December to February. Most eggs recovered from ovitraps belonged to the species Hg. leucocelaenus, representing 85% of all mosquito eggs identified. The other species had lower abundances and percentages: Aedes terrens (7%), Haemagogus janthinomys (5%) and Aedes albopictus (3%). The species that shared breeding sites with a higher frequency were Hg. leucocelaenus and Hg. janthinomys, with a statistically positive correlation (ρ = 0.74). This finding suggests that maybe the presence of Hg. leucocelaenus eggs acted as an attractant for Hg. janthinomys or vice versa. An understanding of mosquito oviposition behavior is necessary for the development of surveillance and control approaches directed against specific pathogen vectors of medical and veterinary importance

Rickettsiae-infected ticks in an endemic area of spotted fever in the State of Minas Gerais, Brazil

A study on tick-borne rickettsiosis was developed in the county of Santa Cruz do Escalvado, State of Minas Gerais, Brazil, where a clinical case of the disease, confirmed by necropsy, had been reported. Of the 1,254 ticks collected, 1,061 belonged to the Amblyomma genus, 57 to the Rhipicephalus sanguineus species, 81 to Boophilus microplus, and 46 to Anocentor nitens. The hemolymph test associated with Gimenez staining showed that 18 of the 221 A. cajennense specimens, 1 of the 16 R. sanguineus, 1 of the 22 B. microplus, 3 of the A. nitens, and 1 of the A. ovale contained rickettsia-like microorganisms. Only 3 A. cajennense ticks were positive under direct immunofluorescence. A. cajennense was the only species found on humans

Early Increase in Myocardial Perfusion After Stem Cell Therapy in Patients Undergoing Incomplete Coronary Artery Bypass Surgery

Incomplete revascularization is associated with worse long-term outcomes. Autologous bone marrow cells (BMC) have recently been tested in patients with severe coronary artery disease. We tested the hypothesis that intramyocardial injection of autologous BMC increases myocardial perfusion in patients undergoing incomplete coronary artery bypass grafting (CABG). Twenty-one patients (19 men), 59 +/- 7 years old, with limiting angina and multivessel coronary artery disease (CAD), not amenable to complete CABG were enrolled. BMC were obtained prior to surgery, and the lymphomonocytic fraction separated by density gradient centrifugation. During surgery, 5 mL containing 2.1 +/- 1.3 x 10(8) BMC (CD34+ = 0.8 +/- 0.3%) were injected in the ischemic non-revascularized myocardium. Myocardial perfusion was assessed by magnetic resonance imaging (MRI) at baseline and 1 month after surgery. The increase in myocardial perfusion was compared between patients with < 50% (group A, n = 11) with that of patients with > 50% (group B, n = 10) of target vessels (stenosis a parts per thousand yenaEuro parts per thousand 70%) successfully bypassed. Injected myocardial segments included the inferior (n = 12), anterior (n = 7), and lateral (n = 2) walls. The number of treated vessels (2.3 +/- 0.8) was significantly smaller than the number of target vessels (4.2 +/- 1.0; P < 0.0001). One month after surgery, cardiac MRI showed a similar reduction (%) in the ischemic score of patients in group A (72.5 +/- 3.2), compared to patients in group B (78.1 +/- 3.2; P = .80). Intramyocardial injection of autologous BMC may help increase myocardial perfusion in patients undergoing incomplete CABG, even in those with fewer target vessels successfully treated. This strategy may be an adjunctive therapy for patients suffering from a more advanced (diffuse) CAD not amenable for complete direct revascularization.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[01/00009-0]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)[552320/2005/6]Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)[552324/2005-1]Fundacao Zerbin

A transitoriedade nos estados de saúde e doença: construção do cotidiano individual e coletivo em uma comunidade rural

O objetivo deste artigo é o de conhecer e compreender as necessidades em saúde, por meio da autopercepção do estado de saúde e doença, considerando as desigualdades sociais presentes na localidade rural de Rincão dos Maia, Canguçu, Rio Grande do Sul. Utilizou-se a triangulação de métodos. No desenho qualitativo entrevistaram-se vinte sujeitos. A amostra foi intencional, ilustrativa das diferentes situações de vida, e a análise, temática. Os resultados indicaram que a sensação de estar adoecido é fortemente vinculada ao modo como se vivencia essa enfermidade, tendo em vista a incapacidade que este estado impõe, seja ela física ou mental. A alternância entre sentir-se doente ou saudável vem da constatação prática de que nenhum estado é contínuo. A observação do meio natural sinaliza o quão tênue são os estados de saúde e doença. A 'situação de vida', elaborada a partir das condições de vida e mobilização de recursos sociais, demonstrou que, apesar das diferenças econômicas, esse grupo apresentava uma coesão social, especialmente em sua matriz cultural de trabalhadores rurais. Enfatiza-se, assim, a necessidade de considerar o significado do fenômeno saúde e doença como ferramenta imprescindível para a formulação de programas de promoção e prevenção à saúde da população rural

Attributable mortality due to nosocomial sepsis in Brazilian hospitals: a case–control study

Abstract Background Nosocomial sepsis is a major healthcare issue, but there are few data on estimates of its attributable mortality. We aimed to estimate attributable mortality fraction (AF) due to nosocomial sepsis. Methods Matched 1:1 case–control study in 37 hospitals in Brazil. Hospitalized patients in participating hospitals were included. Cases were hospital non-survivors and controls were hospital survivors, which were matched by admission type and date of discharge. Exposure was defined as occurrence of nosocomial sepsis, defined as antibiotic prescription plus presence of organ dysfunction attributed to sepsis without an alternative reason for organ failure; alternative definitions were explored. Main outcome measurement was nosocomial sepsis-attributable fractions, estimated using inversed-weight probabilities methods using generalized mixed model considering time-dependency of sepsis occurrence. Results 3588 patients from 37 hospitals were included. Mean age was 63 years and 48.8% were female at birth. 470 sepsis episodes occurred in 388 patients (311 in cases and 77 in control group), with pneumonia being the most common source of infection (44.3%). Average AF for sepsis mortality was 0.076 (95% CI 0.068–0.084) for medical admissions; 0.043 (95% CI 0.032–0.055) for elective surgical admissions; and 0.036 (95% CI 0.017–0.055) for emergency surgeries. In a time-dependent analysis, AF for sepsis rose linearly for medical admissions, reaching close to 0.12 on day 28; AF plateaued earlier for other admission types (0.04 for elective surgery and 0.07 for urgent surgery). Alternative sepsis definitions yield different estimates. Conclusion The impact of nosocomial sepsis on outcome is more pronounced in medical admissions and tends to increase over time. The results, however, are sensitive to sepsis definitions

Association between administration of IL-6 antagonists and mortality among patients hospitalized for COVID-19 : a meta-analysis

IMPORTANCE Clinical trials assessing the efficacy of IL-6 antagonists in patients hospitalized for COVID-19 have variously reported benefit, no effect, and harm. OBJECTIVE To estimate the association between administration of IL-6 antagonists compared with usual care or placebo and 28-day all-cause mortality and other outcomes. DATA SOURCES Trials were identified through systematic searches of electronic databases between October 2020 and January 2021. Searches were not restricted by trial status or language. Additional trials were identified through contact with experts. STUDY SELECTION Eligible trials randomly assigned patients hospitalized for COVID-19 to a group in whom IL-6 antagonists were administered and to a group in whom neither IL-6 antagonists nor any other immunomodulators except corticosteroids were administered. Among 72 potentially eligible trials, 27 (37.5%) met study selection criteria. DATA EXTRACTION AND SYNTHESIS In this prospectivemeta-analysis, risk of biaswas assessed using the Cochrane Risk of Bias Assessment Tool. Inconsistency among trial results was assessed using the I-2 statistic. The primary analysis was an inverse variance-weighted fixed-effects meta-analysis of odds ratios (ORs) for 28-day all-cause mortality. MAIN OUTCOMES AND MEASURES The primary outcome measurewas all-cause mortality at 28 days after randomization. There were 9 secondary outcomes including progression to invasive mechanical ventilation or death and risk of secondary infection by 28 days. RESULTS A total of 10 930 patients (median age, 61 years [range of medians, 52-68 years]; 3560 [33%] were women) participating in 27 trials were included. By 28 days, there were 1407 deaths among 6449 patients randomized to IL-6 antagonists and 1158 deaths among 4481 patients randomized to usual care or placebo (summary OR, 0.86 [95% CI, 0.79-0.95]; P =.003 based on a fixed-effects meta-analysis). This corresponds to an absolute mortality risk of 22% for IL-6 antagonists compared with an assumed mortality risk of 25% for usual care or placebo. The corresponding summary ORs were 0.83 (95% CI, 0.74-0.92; P <.001) for tocilizumab and 1.08 (95% CI, 0.86-1.36; P =.52) for sarilumab. The summary ORs for the association with mortality compared with usual care or placebo in those receiving corticosteroids were 0.77 (95% CI, 0.68-0.87) for tocilizumab and 0.92 (95% CI, 0.61-1.38) for sarilumab. The ORs for the association with progression to invasive mechanical ventilation or death, compared with usual care or placebo, were 0.77 (95% CI, 0.70-0.85) for all IL-6 antagonists, 0.74 (95% CI, 0.66-0.82) for tocilizumab, and 1.00 (95% CI, 0.74-1.34) for sarilumab. Secondary infections by 28 days occurred in 21.9% of patients treated with IL-6 antagonists vs 17.6% of patients treated with usual care or placebo (OR accounting for trial sample sizes, 0.99; 95% CI, 0.85-1.16). CONCLUSIONS AND RELEVANCE In this prospectivemeta-analysis of clinical trials of patients hospitalized for COVID-19, administration of IL-6 antagonists, compared with usual care or placebo, was associated with lower 28-day all-cause mortality

NEOTROPICAL CARNIVORES: a data set on carnivore distribution in the Neotropics

Mammalian carnivores are considered a key group in maintaining ecological health and can indicate potential ecological integrity in landscapes where they occur. Carnivores also hold high conservation value and their habitat requirements can guide management and conservation plans. The order Carnivora has 84 species from 8 families in the Neotropical region: Canidae; Felidae; Mephitidae; Mustelidae; Otariidae; Phocidae; Procyonidae; and Ursidae. Herein, we include published and unpublished data on native terrestrial Neotropical carnivores (Canidae; Felidae; Mephitidae; Mustelidae; Procyonidae; and Ursidae). NEOTROPICAL CARNIVORES is a publicly available data set that includes 99,605 data entries from 35,511 unique georeferenced coordinates. Detection/non-detection and quantitative data were obtained from 1818 to 2018 by researchers, governmental agencies, non-governmental organizations, and private consultants. Data were collected using several methods including camera trapping, museum collections, roadkill, line transect, and opportunistic records. Literature (peer-reviewed and grey literature) from Portuguese, Spanish and English were incorporated in this compilation. Most of the data set consists of detection data entries (n = 79,343; 79.7%) but also includes non-detection data (n = 20,262; 20.3%). Of those, 43.3% also include count data (n = 43,151). The information available in NEOTROPICAL CARNIVORES will contribute to macroecological, ecological, and conservation questions in multiple spatio-temporal perspectives. As carnivores play key roles in trophic interactions, a better understanding of their distribution and habitat requirements are essential to establish conservation management plans and safeguard the future ecological health of Neotropical ecosystems. Our data paper, combined with other large-scale data sets, has great potential to clarify species distribution and related ecological processes within the Neotropics. There are no copyright restrictions and no restriction for using data from this data paper, as long as the data paper is cited as the source of the information used. We also request that users inform us of how they intend to use the data