Serology of Paracoccidioidomycosis Due to Paracoccidioides lutzii

Paracoccidioides lutzii is a new agent of paracoccidioidomycosis (PCM) and has its epicenter localized to the Central-West region of Brazil. Serological diagnosis of PCM caused by P. lutzii has not been established. This study aimed to develop new antigenic preparations from P. lutzii and to apply them in serological techniques to improve the diagnosis of PCM due to P. lutzii. Paracoccidioides lutzii exoantigens, cell free antigen (CFA), and a TCA-precipitated antigen were evaluated in immunodiffusion (ID) tests using a total of 89 patient sera from the Central-West region of Brazil. Seventy-two sera were defined as reactive for P. brasiliensis using traditional antigens (AgPbB339 and gp43). Non-reactive sera for traditional antigens (n = 17) were tested with different P. lutzii preparations and P. lutzii CFA showed 100% reactivity. ELISA was found to be a very useful test to titer anti-P. lutzii antibodies using P. lutzii-CFA preparations. Sera from patients with PCM due to P. lutzii presented with higher antibody titers than PCM due to P. brasiliensis and heterologous sera. in western blot, sera from patients with PCM due to P. lutzii were able to recognize antigenic molecules from the P. lutzii-CFA antigen, but sera from patients with PCM due to P. brasiliensis could not recognize any P. lutzii molecules. Due to the facility of preparing P. lutzii CFA antigens we recommend its use in immunodiffusion tests for the diagnosis of PCM due to P. lutzii. ELISA and western blot can be used as complementary tests.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo UNIFESP, Dept Microbiol Imunol & Parasitol, Disciplina Biol Celular, São Paulo, BrazilUniv Fed Mato Grosso do Sul UFMS, Fac Med FAMED, Campo Grande, MS, BrazilUniv Fed Mato Grosso UFMT, Nucleo Doencas Infecciosas & Trop, Cuiaba, Mato Grosso, BrazilUniversidade Federal de São Paulo UNIFESP, Dept Microbiol Imunol & Parasitol, Disciplina Biol Celular, São Paulo, BrazilFAPESP: 2012/06593-0FAPESP: 2009/54024-2Web of Scienc

Diagnóstico de autismo infantil e suas repercussões nas relações familiares e educacionais/ Diagnosis of childhood autism and its repercussions on family and educational relationships

Os transtornos do espectro autista (TEAs) são vitalícios e normalmente são doenças devastadoras que afetam gravemente o funcionamento social e a autossuficiência. O objetivo desse trabalho é compreender a inserção do indivíduo autista na família e no ambiente escolar e seus efeitos. Inicialmente, 32 artigos foram selecionados das bases Medical Literature Analysis and Retrieval System Online (MedLine), Public Medline (PubMed) e Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs). Posteriormente, foram selecionados os artigos confeccionados a partir de 2015, com foco temático na repercussão do autismo na qualidade de vida dos pais e na vida escolar dessas crianças. São 20 artigos no total. O diagnóstico de autismo em si, é uma quebra de expectativas de futuro para os pais. Não somente, com o passar dos anos causa alterações na estrutura familiar, já que esta deve sempre auxiliar o indivíduo até em tarefas simples. Isso pode acarretar estresse e depressão nos pais. Ademais, preconceito e exclusão no meio escolar são recorrentes pela falta de compreensão da doença pela sociedade. Envolta em vários questionamentos, os transtornos do espectro autista devem ser cada vez mais estudados, com o principal objetivo de melhorar a qualidade de vida das famílias afetadas

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Forensic electrochemistry: Electrochemical study and quantification of xylazine in pharmaceutical and urine samples

Xylazine is marketed as a veterinary drug and used for sedative and analgesic purposes; it is not approved for human use due to various side effects. In recent years, xylazine has emerged as a drug of abuse and has been associated with drug-facilitated sexual assaults. Herein, we describe, for the first time, the electrochemical study of xylazine in aqueous and organic media and proposed a quantification method using glassy carbon electrode. The simple and accurate quantification method was performed using differential pulse voltammetry. Under the optimized experimental conditions, a linear analytical curve was obtained for xylazine concentrations ranging from 0.5 to 256 mu mol L-1, with an estimated detection limit of 120 nmol L-1. The proposed method was applied to determination of xylazine in pharmaceutical formulation. Furthermore, to forensic scenario, a standard addition technique on urine samples exhibited recovery values in the range of 93-103%, demonstrating the potentiality to forensic and clinical analysis of xylazine intoxication295726734CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP306504-2011-1; 2012/06642-1; EMU 2015/08539-13359/2014 PRO-FORENSES Edital 25/20142016/16477-9; 2017/10522-

A) Western blot assays showing the reactivity of sera from patients with PCM due to <i>P. lutzii</i> versus <i>P. lutzii</i>-CFA antigen.

<p>Hc, Asp, Sp and NHS: representative blots of heterologous sera and normal human sera (15 sera each). (*) = represents <i>P. lutzii</i> gold standard serum. <b>B</b>) Reactivity of sera from patients with PCM due to <i>P. brasiliensis</i> versus <i>P. lutzii</i>-CFA antigen. We could distinguish <i>P. brasiliensis</i> from <i>P. lutzii</i> sera, as only sera from patients with PCM due to <i>P. lutzii</i> recognize antigenic molecules from <i>P. lutzii</i>-CFA. <b>C</b>) Western blot assays showing the reactivity of sera from patients with PCM due to <i>P. brasiliensis</i> versus <i>P. brasiliensis</i>-CFA (strain B339). Gp43 is basically recognized by PCM sera due to <i>P. brasiliensis</i>. <b>D</b>) Reactivity of sera due to <i>P. lutzii</i> versus <i>P. brasiliensis</i>-CFA (strain B339).</p

Electron microscopy of <i>P. lutzii</i> yeast phase showing multiple budding cells.

<p>Electron microscopy of <i>P. lutzii</i> yeast phase showing multiple budding cells.</p

Receiver operating characteristics (ROC) curve for the CFA, EXO and TCA antigens derived from <i>Paracoccidioides lutzii</i> EPM208 isolate.

<p>The ROC is plotted between the true-positive rate (sensitivity) on the y-axis, and the false-positive rate (100-specificity) on the x-axis. The area under the curve (AUC) represents the accuracy of the ID test which was 1.0, IC 0.95–1.0, p<0.0001 for the CFA, 0.74±0.06, IC 0.68–0.87, p<0.0001 for the EXO and 0.58±0.04, IC 0.47–0.69, p = 0.0641 for the TCA antigens. The higher AUC above 0.5, the better the test.</p

<i>Paracoccidioides lutzii</i> isolates used in the study.

<p>*EPM = Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.</p

A) Individual serum ELISA titers (1/T) of 28 sera from patients with PCM due to <i>P. lutzii</i> (blue) compared to individual serum ELISA titers (1/T) of 28 sera from patients with PCM due to <i>P. brasiliensis</i> (red) reacting with Ag CFA-Pl EPM 208 (12.5 µg/ml).

<p>The <i>P. lutzii</i> sera presented ID + for <i>P. lutzii</i> CFA antigen and ID − for <i>P. brasiliensis</i> exoantigen (AgPbB339). The group of <i>P. brasiliensis</i> sera presented ID + for AgPbB339 antigen and ID − for <i>P. lutzii</i> CFA antigen. (<b>*</b>) represents <i>P. lutzii</i> gold standard serum and (<b>**</b>) represents sera that had a very weak cross reaction with <i>P. brasiliensis</i> exoantigen. <b>B</b>) Individual serum ELISA titers (1/T) of 28 sera from patients with PCM due to <i>P. lutzii</i> (blue) compared to individual serum ELISA titers (1/T) of 28 sera from patients with PCM due to <i>P. brasiliensis</i> (red) reacting with Ag Exo Pb B339 (10 µg/ml). The <i>P. lutzii</i> sera presented ID + for <i>P. lutzii</i> CFA antigen and ID − for <i>P. brasiliensis</i> exoantigen (AgPbB339). The group of <i>P. brasiliensis</i> sera presented ID + for AgPbB339 antigen and ID − for <i>P. lutzii</i> CFA antigen. (*) represents <i>P. lutzii</i> gold standard serum and (**) represents sera that had a very weak cross reaction with <i>P. brasiliensis</i> exoantigen. The same group of sera was used in A and B. <b>C</b>) ELISA median curves from sera from patients with PCM due to <i>P. lutzii</i>, PCM due to <i>P. brasiliensis</i>, and heterologous sera such as histoplasmosis, aspergillosis, sporotrichosis, and normal human serum. Antigen used was <i>P. lutzii</i> CFA (Ag CFA-Pl EPM 208 at 12.5 µg/ml). <b>D</b>) ELISA individual serum titers (1/T) of patients with PCM due to <i>P. lutzii</i>, <i>P. brasiliensis</i>, and heterologous sera such as Histoplasmosis, Aspergillosis, Sporotrichosis and normal human sera. By the end titer we could distinguish sera of PCM patients due to <i>P. lutzii</i> from sera of PCM due to <i>P. brasiliensis</i> and other mycotic diseases. Antigen used was <i>P. lutzii</i> CFA (Ag CFA-Pl EPM 208 at 12.5 µg/ml). Bars indicate the median.</p

A) Immunodiffusion tests with different CFAs from 12 strains of <i>Paracoccidioides lutzii</i>.

<p> 1 = EPM 147; 2 = EPM 148; 3 = EPM 196; 4 = EPM 201; 5 = EPM 205; 6 = EPM 206; 7 = EPM 208; 8 = EPM 212; 9 = EPM 213; 10 = EPM 223; 11 = EPM 209; 12 = EPM 226. In the center well, <b>S*</b> = <i>P. lutzii</i> serum (gold standard). <i>P. lutzii</i> strains EPM 227 and EPM 228 were positive (not shown). <b>B</b>) Immunodiffusion test showing that <b>S*</b> = <i>P. lutzii</i> gold standard serum do not react with CFA from <i>P. brasiliensis</i> Pb18 and B339. <b>C</b>) Immunodiffusion test using Ag Exo (crude exoantigen of <i>P. lutzii</i>) and Ag TCA (crude exoantigen of <i>P. lutzii</i> precipitated with trichloroacetic acid) versus <i>P. lutzii</i> serum (<b>S*</b> = <i>P. lutzii</i> gold standard serum and S = suspected <i>P. lutzii</i> serum). Observe the weak band in both cases. <b>D</b>) immunodiffusion test showing that S* (<i>P. lutzii</i> gold standard serum) do not react with Ag Exo, Ag TCA and CFA from <i>P. brasiliensis</i> B339 antigen. <b>E</b>) Immunodiffusion test of 12 sera from patients with PCM due to <i>P. lutzii</i> using <i>P. lutzii</i> CFA. In the center well is the CFA antigen (Ag), and different PCM sera are in the peripheral wells (1 to 12). <b>F</b>) Immunodiffusion test showing that <b>S*</b> (<i>P. lutzii</i> gold standard serum) do not react with Exo or CFA of <i>P. brasiliensis</i> B339; and S_Pb (serum of PCM patient due to <i>P. brasiliensis</i>) do not react with Exo or CFA of <i>P. lutzii</i>.</p