Identification of a 17 kDa protein that is a potentially novel antigen of lettuce-associated respiratory allergy in farmers.

BACKGROUND: We have identified and reported a novel antigen, nonprotein-specific secreted EP1-like glycoprotein (51 kDa), for lettuce-related respiratory allergy. OBJECTIVE: We aimed to identify a novel antigen for lettuce-related respiratory allergy that is different from epidermis-specific secreted EP1-like glycoprotein. METHODS: Immunoblotting was performed using an immunoglobulin E-specific antibody. The antigen-antibody reaction was confirmed by means of enzyme-linked immunosorbent assaying. LC-MS/MS analysis was carried out to detect a novel protein found in sera from 3 of 13 patients with lettuce-related respiratory allergy. Finally, we purified a novel protein from Escherichia coli. RESULTS: Immunoblotting assays showed common bands of 17 kDa in the sera of 3 of 13 patients. An enzyme-linked immunosorbent assay confirmed that the patient sera reacted with lettuce latex juice. A 17 kDa protein band that showed antigenic reactivity in 3 of 13 patient sera was identified as a kirola-like protein by LC-MS/MS. In addition, although we purified this protein, we failed to show the inhibitory effect. CONCLUSION: A 17 kDa protein that is a potentially novel antigen of lettuce-associated respiratory allergy was identified. In further studies, we will focus on purifying this novel protein to diagnose lettuce allergy.

Fast and cost-effective protocol to produce Paracoccidioides spp. antigens / Protocolo rápido y económico para la producción de antígenos de Paracoccidioides spp

INTRODUCTION: The existing methods for Paracoccidioides spp. antigen production are problematic in terms of standardization, specificity, stability, repeatability, and reproducibility. OBJECTIVE: To optimize the methodology for Paracoccidioides spp. antigen production and evaluate its applicability in paracoccidioidomycosis immunodiagnosis. MATERIALS AND METHODS: The antigens were obtained from Paracoccidioides lutzii isolates (01, 66, and 8334), Paracoccidioides brasiliensis sensu stricto (113), and Paracoccidioides restripiensis (B-339). These fungi were grown at 36 °C ± 1 °C, on modified Fava-Netto agar, according to Freitas et al. (2018). Paracoccidioides lutzii antigens were obtained after , 10, and 20 days of culture, whereas P. brasiliensis and P. restripiensis antigens were obtained after 10 days. Antigens were evaluated in natura, 10 and 20 times concentrated. Antigenic capacity was evaluated using a double immunodiffusion assay against serum samples from patients with paracoccidioidomycosis, histoplasmosis, and aspergillosis, and random blood donors. RESULTS: Cross-reactivity between Paracoccidioides spp. antigens was observed when P. brasiliensis, P. restrepiensis antigens, and P. lutzii antigens were evaluated with the polyclonal antibodies against P. lutzii and P. brasiliensis, respectively. No cross-reactivity was obtained for polyclonal antibodies against Histoplasma capsulatum, Aspergillus fumigatus, and random blood donors. The proposed protocol allowed stable, repeatable, and reproducible genus-specific antigen production at a low cost and in a short cultivation time. CONCLUSION: The proposed protocol allowed us to obtain genus-specific antigens that can be developed and reproduced in all laboratories in Brazil and South America, where paracoccidioidomycosis is a neglected disease, contributing to an early diagnosis, especially in endemic regions, regardless of the species.

Introducción: Los métodos existentes para la producción de los antígenos de Paracoccidioides spp. son problemáticos en su estandarización, especificidad, estabilidad, repetibilidad y reproducibilidad. Objetivo: Optimizar la metodología para la producción de antígenos de Paracoccidioides spp. y evaluar su aplicabilidad en el inmunodiagnóstico de la paracoccidioidomicosis. Materiales y métodos: Los antígenos se obtuvieron de aislamientos de P. lutzii (01, 66 y 8334), P. brasiliensis sensu stricto (113) y P. restripiensis (B-339). Estos hongos se cultivaron a 36 °C ± 1 °C en agar Fava-Netto modificado, según Freitas et al. (2018). Los antígenos de P. lutzii se obtuvieron a los 5, 10 y 20 días de cultivo y los antígenos de P. brasiliensis y P. restripiensis se obtuvieron a los 10 días. Los antígenos se evaluaron in natura, concentrados 10 y 20 veces. La capacidad antigénica se evaluó mediante un ensayo de inmunodifusión doble con muestras de suero de pacientes con paracoccidioidomicosis, histoplasmosis, aspergilosis y donantes de sangre aleatorios. Resultados: Se observó reacción cruzada con Paracoccidioides spp. cuando se evaluaron los antígenos de P. brasiliensis, P. restrepiensis y P. lutzii frente a los anticuerpos policlonales contra P. lutzii y P. brasiliensis, respectivamente. No hubo reactividad cruzada con los anticuerpos policlonales contra Histoplasma capsulatum y Aspergillus fumigatus, ni contra los donantes de sangre aleatorios. El protocolo propuesto permitió la producción estable, repetible y reproducible de antígenos dirigidos de un género específico (Paracoccidiodes) en un tiempo corto de cultivo y a un menor costo. Conclusión: El protocolo propuesto permitió obtener antígenos específicos de un género, que pueden ser desarrollados y reproducidos en todos los laboratorios de Brasil y Surámerica donde la paracoccidioidomicosis es una enfermedad endémica y desatendida. Estos antígenos pueden contribuir al diagnóstico precoz de la infección, independientemente de la especie.

Fast and cost-effective protocol to produce Paracoccidioides spp. antigens / Protocolo rápido y económico para la producción de antígenos de Paracoccidioides spp

Introduction. The existing methods for Paracoccidioides spp. antigen production are problematic in terms of standardization, specificity, stability, repeatability, and reproducibility. Objective. To optimize the methodology for Paracoccidioides spp. antigen production and evaluate its applicability in paracoccidioidomycosis immunodiagnosis. Materials and methods. The antigens were obtained from Paracoccidioides lutzii isolates (01, 66, and 8334), Paracoccidioides brasiliensis sensu stricto (113), and Paracoccidioides restripiensis (B-339). These fungi were grown at 36 °C ± 1 °C, on modified Fava-Netto agar, according to Freitas et al. (2018). Paracoccidioides lutzii antigens were obtained after 5, 10, and 20 days of culture, whereas P. brasiliensis and P. restripiensis antigens were obtained after 10 days. Antigens were evaluated in natura, 10 and 20 times concentrated. Antigenic capacity was evaluated using a double immunodiffusion assay against serum samples from patients with paracoccidioidomycosis, histoplasmosis, and aspergillosis, and random blood donors. Results. Cross-reactivity between Paracoccidioides spp. antigens was observed when P. brasiliensis, P. restrepiensis antigens, and P. lutzii antigens were evaluated with the polyclonal antibodies against P. lutzii and P. brasiliensis, respectively. No cross-reactivity was obtained for polyclonal antibodies against Histoplasma capsulatum, Aspergillus fumigatus, and random blood donors. The proposed protocol allowed stable, repeatable, and reproducible genus-specific antigen production at a low cost and in a short cultivation time. Conclusion. The proposed protocol allowed us to obtain genus-specific antigens that can be developed and reproduced in all laboratories in Brazil and South America, where paracoccidioidomycosis is a neglected disease, contributing to an early diagnosis, especially in endemic regions, regardless of the species.

Introducción. Los métodos existentes para la producción de los antígenos de Paracoccidioides spp. son problemáticos en su estandarización, especificidad, estabilidad, repetibilidad y reproducibilidad. Objetivo. Optimizar la metodología para la producción de antígenos de Paracoccidioides spp. y evaluar su aplicabilidad en el inmunodiagnóstico de la paracoccidioidomicosis. Materiales y métodos. Los antígenos se obtuvieron de aislamientos de P. lutzii (01, 66 y 8334), P. brasiliensis sensu stricto (113) y P. restripiensis (B-339). Estos hongos se cultivaron a 36 °C ± 1 °C en agar Fava-Netto modificado, según Freitas et al. (2018). Los antígenos de P. lutzii se obtuvieron a los 5, 10 y 20 días de cultivo y los antígenos de P. brasiliensis y P. restripiensis se obtuvieron a los 10 días. Los antígenos se evaluaron in natura, concentrados 10 y 20 veces. La capacidad antigénica se evaluó mediante un ensayo de inmunodifusión doble con muestras de suero de pacientes con paracoccidioidomicosis, histoplasmosis, aspergilosis y donantes de sangre aleatorios. Resultados. Se observó reacción cruzada con Paracoccidioides spp. cuando se evaluaron los antígenos de P. brasiliensis, P. restrepiensis y P. lutzii frente a los anticuerpos policlonales contra P. lutzii y P. brasiliensis, respectivamente. No hubo reactividad cruzada con los anticuerpos policlonales contra Histoplasma capsulatum y Aspergillus fumigatus, ni contra los donantes de sangre aleatorios. El protocolo propuesto permitió la producción estable, repetible y reproducible de antígenos dirigidos de un género específico (Paracoccidiodes) en un tiempo corto de cultivo y a un menor costo. Conclusión. El protocolo propuesto permitió obtener antígenos específicos de un género, que pueden ser desarrollados y reproducidos en todos los laboratorios de Antígenos de Paracoccidioides spp.: protocolo rápido Brasil y Surámerica donde la paracoccidioidomicosis es una enfermedad endémica y desatendida. Estos antígenos pueden contribuir al diagnóstico precoz de la infección, independientemente de la especie.

Detection of Serum-Specific IgE by Fluoro-Enzyme Immunoassay for Diagnosing Type I Hypersensitivity Reactions to Penicillins.

Diagnosis of type I hypersensitivity reactions (IgE-mediated reactions) to penicillins is based on clinical history, skin tests (STs), and drug provocation tests (DPTs). Among in vitro complementary tests, the fluoro-enzyme immunoassay (FEIA) ImmunoCAP® (Thermo-Fisher, Waltham, MA, USA) is the most widely used commercial method for detecting drug-specific IgE (sIgE). In this study, we aimed to analyze the utility of ImmunoCAP® for detecting sIgE to penicillin G (PG) and amoxicillin (AX) in patients with confirmed penicillin allergy. The study includes 139 and 250 patients evaluated in Spain and Italy, respectively. All had experienced type I hypersensitivity reactions to penicillins confirmed by positive STs. Additionally, selective or cross-reactive reactions were confirmed by DPTs in a subgroup of patients for further analysis. Positive ImmunoCAP® results were 39.6% for PG and/or AX in Spanish subjects and 52.4% in Italian subjects. When only PG or AX sIgE where analyzed, the percentages were 15.1% and 30.4%, respectively, in Spanish patients; and 38.9% and 46% in Italian ones. The analysis of positive STs showed a statistically significant higher percentage of positive STs to PG determinants in Italian patients. False-positive results to PG (16%) were detected in selective AX patients with confirmed PG tolerance. Low and variable sensitivity values observed in a well-defined population with confirmed allergy diagnosis, as well as false-positive results to PG, suggest that ImmunoCAP® is a diagnostic tool with relevant limitations in the evaluation of subjects with type I hypersensitivity reactions to penicillins.

Early infant diagnosis of HIV infection at the John F. Kennedy Medical Center, Monrovia, Liberia.

Background: Vertical transmission accounts for majority of new HIV infections among children worldwide. Ninety percent of HIV-positive children reside in Sub- Saharan Africa with their infection predominantly acquired via vertical transmission. In 2004, the vertical transmission rate of HIV in Africa was estimated at 25 - 40% but, remarkably, the rate has significantly decreased to less than 5% in most African countries following implementation and expansion of prevention of MTCT (PMTCT) programs.Objective: To determine the rate of and factors associated with vertical transmission of HIV among attendees of early infant diagnosis (EID) program of an academic and community-based tertiary facility in Liberia.Design: A retrospective cross-sectional analysis.Methods: A retrospective review of medical records of babies seen at Pediatric Unit of Infectious Disease Clinic of John F Kennedy Medical Center (JFKMC) in Monrovia, Liberia between January 1, 2016 and December 31, 2020. All subjects were children born to HIV-positive mothers and who had HIV DNA PCR testing performed between the ages of 6 weeks and 6 months. Children who suffered early neonatal death and those who did not undergo PCR testing were excluded. Demographics of mother to child pairs as well as factors known to influence vertical transmission of HIV such as partial (15.8%) or full (84.2%) participation in prevention of MTCT (PMTCT) programs, mode of delivery, breastfeeding and utilization of post-exposure prophylaxis were collected and assessed. Binomial logistic regression analyses were used to assess factors associated with vertical transmission.Results: During the study timeframe, 284 children had a HIV DNA PCR test with a male:female ratio - 1.3:1. Sixteen tested positive (conducted at a mean of 155 days post birth) giving a vertical transmission rate of 5.6%. For 239 mothers (84.2%) who had full PMTCT, 1.3% of their children tested positive, while for 45 mothers (15.8%) who had partial PMTCT, 28.8% of their children being positive. Two hundred and seventy six children (97%) had exclusive breastfeeding, 13 of whom tested positive while 2 children who were mixed fed tested positive. Children who had Nevirapine vs no prophylaxis (OR = 1.89[95% CI 1.16 - 2.96]), were delivered via caesarian section vs vaginal delivery (OR= 2.26[95% CI 1.92 - 4.12].) and full versus partial participation in PMTCT programs (OR = 4.02[95% CI 2.06 - 4.13] were more likely to have negative HIV test.Conclusion: Vertical transmission rate was found to be high in Liberia and may be driven by suboptimal PMTCT program participation including post-exposure prophylaxis for infants. Therefore, strategies to scale up and improve uptake of PMTCT services are needed to mitigate the burden of HIV among children.

Multiparametric assay of screening for the diagnosis of mycoses of interest in Public Health: standardization of methodology / Ensaio multiparamétrico de triagem para diagnóstico de micoses de interesse em Saúde Pública: padronização da metodologia

The standardization and validation of a multiplex assay requires the combination of important parameters such as sensitivity and specificity, acceptable levels of performance, robustness, and reproducibility. We standardized a multiparametric Dot-blot aimed at the serological screening of paracoccidioidomycosis, histoplasmosis, and aspergillosis. A total of 148 serum were evaluated: 10 from healthy subjects, 36 from patients with paracoccidioidomycosis, 62 from patients with histoplasmosis, and 40 from patients with aspergillosis. It was found that the multiparametric Dot-blot showed a high percentage of cross-reactivity. However, when evaluated individually, in the serological screening of histoplasmosis, a good performance was observed when compared to the double immunodiffusion assay, considered the gold standard test, with 100% co-positivity and 83.3% co-negativity. The performance of serological screening for aspergillosis was not satisfactory when compared to double immunodiffusion, showing 71.4% co-positivity and 100% co-negativity. The evaluation of the stability of nitrocellulose membranes showed that membranes sensitized with H. capsulatum antigen remained stable for 90 days and those sensitized with A. fumigatus antigen for 30 days. We conclude that the use of crude antigens was not suitable for the standardization of the multiparametric Dot-blot assay, due to the high cross-reactivity, and that further tests should be performed with purified proteins (AU).

A padronização e validação de um ensaio multiplex requer a combinação de parâmetros importantes, como sensibilidade e especificidade, níveis aceitáveis de desempenho, robustez e reprodutibilidade. Este trabalho padronizou um Dot-blot multiparamétrico visando a triagem sorológica da paracoccidioidomicose, histoplasmose e aspergilose. Foram avaliadas 148 amostras de soro: 10 de indivíduos saudáveis, 36 de pacientes com paracoccidioidomicose, 62 de pacientes com histoplasmose e 40 de pacientes com aspergilose. Verificou-se que o Dot-blot multiparamétrico apresentou elevado percentual de reatividade cruzada. Entretanto, quando avaliado individualmente, na triagem sorológica da histoplasmose observou-se bom desempenho quando comparado ao ensaio de imunodifusão dupla, considerado o teste padrão ouro, com 100% de co-positividade e 83,3% de co-negatividade. O desempenho da triagem sorológica da aspergilose não foi satisfatório quando comparado a imunodifusão dupla, apresentando 71,4% de co-positividade e 100% de co-negatividade. A avaliação da estabilidade das membranas de nitrocelulose mostrou que membranas sensibilizadas com antígeno de H. capsulatum permaneceram estáveis por 90 dias e as sensibilizadas com antígeno de A. fumigatus, por 30 dias. Concluímos que o uso de antígenos brutos não foi adequado para a padronização do ensaio de Dot-blot multiparamétrico, devido ao alto índice de reatividade cruzada, e que novos testes devem ser realizados com proteínas purificadas (AU).

Severe cutaneous adverse drug reactions: diagnostic approach and genetic study in a Brazilian case series.

Summary: Background. Severe cutaneous adverse reactions (SCAR) are potentially fatal reactions. Genetic predisposition is involved in their pathogenesis related to drugs and ethnicities, however in a mixed population these relationships are still unknown. The aim of this study was to describe phenotypes, suspect drugs and HLA-alleles related to SCAR, identified by a systematized approach in a Brazilian case series. Methods. Patients who were diagnosed with SCAR between March 2011 and July 2019 at our university hospital were included. European Network for Drug Allergy (ENDA) questionnaire was used to collect clinical and laboratory data and algorithms for assessment of drug causality were applied. Socio-demographic variables included age, gender and skin color/ethnicity. Drug patch tests (DPT) and HLA-A, -B, -DRB1 typing were carried out. Results. A total of 74 patients were included: 36 (48.64%) with SJS/TEN, 32 (43.24%) DRESS/DIHS, 3 (4.05%) AGEP, 2 (2.70%) overlap(DRESS/SJS and DRESS/AGEP) and 1 (1.35%) GBFDE. The median age was31.5 years (IQR = 14-52.25), most were female (n = 44/59.46%) and brown (n = 38/51.35%). Anticonvulsants (n = 32/43.24%) were the largest group involved and antibiotics (n = 26/35.13%) were the second most common. Two patients with DRESS died during the acute phase. Positive DPT were shown only in anticonvulsant associated DRESS. HLA related to abacavir, allopurinol and carbamazepine were identified. Conclusions. A systematized approach allowed the phenotypic characterization of SCAR. The HLA-A*31:01, B*57:01 and B*58:01 alleles were identified, reinforcing the causality in SCAR by CBZ, ABC and ALLO in the Brazilian population.

Long COVID following mild SARS-CoV-2 infection: characteristic T cell alterations and response to antihistamines.

Long COVID is characterized by the emergence of multiple debilitating symptoms following SARS-CoV-2 infection. Its etiology is unclear and it often follows a mild acute illness. Anecdotal reports of gradual clinical responses to histamine receptor antagonists (HRAs) suggest a histamine-dependent mechanism that is distinct from anaphylaxis, possibly mediated by T cells, which are also regulated by histamine. T cell perturbations have been previously reported in post-viral syndromes, but the T cell landscape in patients who have recovered from mild COVID-19 and its relationship to both long COVID symptoms and any symptomatic response to HRA remain underexplored. We addressed these questions in an observational study of 65 individuals who had recovered from mild COVID-19. Participants were surveyed between 87 and 408 days after the onset of acute symptoms; none had required hospitalization, 16 had recovered uneventfully, and 49 had developed long COVID. Symptoms were quantified using a structured questionnaire and T cell subsets enumerated in a standard diagnostic assay. Patients with long-COVID had reduced CD4+ and CD8+ effector memory (EM) cell numbers and increased PD-1 (programmed cell death protein 1) expression on central memory (CM) cells, whereas the asymptomatic participants had reduced CD8+ EM cells only and increased CD28 expression on CM cells. 72% of patients with long COVID who received HRA reported clinical improvement, although T cell profiling did not clearly distinguish those who responded to HRA. This study demonstrates that T cell perturbations persist for several months after mild COVID-19 and are associated with long COVID symptoms.

Evaluation of skin test indications for general anesthetics in real life: a prospective cohort study.

BACKGROUND: In daily practice, atopic patients and those who have other drug allergies are referred to allergy clinics for evaluation of possible general anesthetic allergy despite the fact that it is not recommended in recent guidelines. OBJECTIVE: The aim of this prospective study is to determine the negative predictive value of skin tests for common general anesthetic drugs prior to general anesthesia in atopic patients and in patients who had drug allergies by including the data of those who had previously tolerated or reacted to general anesthesia. METHODS: A database program was constituted to collect the preoperative skin test data of patients referred to our clinic between 2013 and 2018. Demographic and clinical history, medications implemented during perioperative period, reactions, and results of skin tests performed with anesthetic drugs and latex were evaluated. RESULTS: Four hundred fifty-nine out of the total 1167 patients referred fulfilled the inclusion criteria for further evaluation. Nearly 75% of the patients were female and mean age was 46.3 ±â€¯14.3 years. History of hypersensitivity reactions (HRs) due to NSAIDs and/or antibiotics, radiocontrast agents, local anesthetics, and food were present in the 53.1%, 4.1%, 1.5%, and 2.0%, respectively. The negative predictive values of skin tests for general anesthetics were in the range of 80-100%. Only 4 patients (0,87%) experienced HRs during operation. CONCLUSION: These real-life data reveal high rates of negative predictive value of skin tests with general anesthetic drugs and a low reaction rate in atopic patients and in patients with allergy to other drugs.

Current status and research progress of microfluidic immunochips in medical detection / 中华检验医学杂志

The traditional-immunological strategies for clinical laboratories often rely on large and expensive instruments and skilled operators, and the measurement time is also long. However, the sensitivity of these strategies is still unsatisfactory. It is urgent to research and develop the point-of-care testing (POCT) featured as a highly sensitive, accurate, and rapid/POCT diagnosis. The Microfluidic chips have multi-advantages that are suitable for the clinical POCT diagnosis: high sensitivity, throughput, and automation. Recently, the Microfluidic-immune chips developed based on the microfluidic technology combined with immune detection have considered not only hotspots in the related research but also benefit to the tumor marker detection, antigen and antibody detection of infectious diseases, autoantibody detection, hormone detection, and other fields. However, there are still many challenges to be overcome during the application of chips, such as more effective microfluidic manipulation, more sensitive collection, and analysis of reaction signals.

[In current practice, what is the best test to detect blood in the stool ?] / En pratique courante, quel est le meilleur test pour rechercher du sang dans les selles ?

"In current practice, what is the best test to detect the presence of blood in faeces? Testing for the presence of blood in faeces is still widely performed in France, although the uptake in the national colorectal cancer screening programme is less than desirable, as is use in routine practice to guide diagnosis when patients present with gastrointestinal or general symptoms. Guaiac-based tests (Hemoccult) are obsolete and have been replaced by faecal immunochemical tests (FITs), both quantitative (OC-Sensor), currently reserved for the screening programme, and qualitative, performed by all medical laboratories. However, qualitative FITs should be abandoned: their visual reading is subjective, their positivity rate is highly variable, around 50%, and their diagnostic sensitivity for colorectal cancer has not been well determined. In contrast, quantitative FITs are reliable and their performance has been well evaluated. Several neighbouring countries use these tests in general practice to assess the risk of colorectal cancer in symptomatic patients and to determine the indication for colonoscopy. In this context, the decision thresholds are very different from those of organised screening: a faecal haemoglobin concentration below 4-10 µg/g is associated with a very low risk of cancer, a concentration above 4-10 µg/g should be investigated by colonoscopy, and a concentration above 150 µg/g requires an urgent colonoscopy."

"En pratique courante, quel est le meilleur test pour rechercher du sang dans les selles ? La recherche de sang dans les selles, malgré son caractère quelque peu désuet, est, à juste titre, encore très prescrite en France, insuffisamment dans le programme national de dépistage organisé du cancer colorectal et en pratique courante pour guider une conduite diagnostique en cas de symptômes digestifs ou généraux. Les tests au gaïac (Hemoccult), obsolètes, ont été remplacés par des tests immunochimiques, quantitatifs (OC-Sensor) réservés au programme de dépistage, et qualitatifs réalisés par tous les laboratoires de biologie médicale. Pourtant les tests qualitatifs sont obsolètes et devraient être abandonnés : leur lecture est subjective, leur taux de positivité très variable, proche de 50 %, et leur sensibilité diagnostique pour le cancer colorectal non évaluée. Au contraire, les tests quantitatifs sont fiables et leurs performances parfaitement évaluées. Plusieurs pays voisins utilisent ces tests en médecine générale pour évaluer le risque de cancer colorectal chez les patients symptomatiques et poser l'indication d'une coloscopie. Dans ce contexte, les seuils décisionnels sont très différents de ceux du dépistage organisé : un taux d'hémoglobine fécale inférieur à 4-10 µg/g de selles est associé à un risque infime de cancer, un taux supérieur à 4-10 µg/g doit être exploré par coloscopie, et un taux supérieur à 150 µg/g relève d'une coloscopie urgente."

Performance of Antigen-Based Testing as Frontline Diagnosis of Symptomatic COVID-19.

Background and Objectives: To evaluate the performance of antigen-based detection tests as the frontline diagnosis of coronavirus disease 2019 (COVID-19). Materials and Methods: We conducted a nationwide retrospective cohort study in Mexico. A cross-sectional analysis of a cohort study was conducted in Mexico and data from 15,408 suspected (all of them symptomatic) cases of COVID-19 were analyzed. The results of antigen-based tests were compared with those obtained by molecular (polymerase chain reaction-based) assays. Results: The antigen-based tests showed sensitivity below 50% and high specificity in all the analyzed age groups. The highest Youden index (J) was observed among adults aged 25-44 years old (45.5, 95% CI 43.7-47.3). Conclusions: We documented the poor performance of serologic techniques as frontline diagnosis of symptomatic COVID-19 and inaccurate results may impact negatively on pandemic progression.

Flipped Quick-Response Code Enables Reliable Blood Grouping.

Accurate and rapid blood typing plays a vital role in a variety of biomedical and forensic scenarios, but recognizing weak agglutination remains challenging. Herein, we demonstrated a flipping identification with a prompt error-discrimination (FLIPPED) platform for automatic blood group readouts. Bromocresol green dye was exploited as a characteristic chromatography indicator for the differentiation of plasma from whole blood by presenting a teal color against a brown color. After integrating these color changes into a quick-response (QR) code, prompt typing of ABO and Rhesus groups was automatically achieved and data could be uploaded wirelessly within 30 s using a commercially available smartphone to facilitate blood cross-matching. We further designed a color correction model and algorithm to remove potential errors from scanning angles and ambient light intensities, by which weak agglutination could be accurately recognized. With comparable accuracy and repeatability to classical column assay in grouping 450 blood samples, the proposed approach further demonstrates to be a versatile sample-to-result platform for clinical diagnostics, food safety, and environmental monitoring.

Analysis of the spatio-temporal dynamics of incidence, mortality and test rates (rapid and RT-PCR) of COVID-19 in the state of Minas Gerais, Brazil / Análise da dinâmica espaço-temporal da incidência, mortalidade e taxas de testes (rápidos e moleculares) da COVID-19 no estado de Minas Gerais, Brasil / Análisis de la dinámica espacio-temporal de la incidencia, mortalidad y tasas de prueba (rápida e RT-PCR) de COVID-19 en el estado de Minas Gerais, Brasil

Background and Objectives: A novel type of coronavirus, SARS-CoV-2, is responsible for an unprecedented pandemic with profound socioeconomic consequences. Owing to its recent discovery still represents a great unknown to researchers. Thus, this study aims to establish the spatio-temporal associations of the incidence, mortality, and the rate of both rapid and RT-PCR tests in Minas Gerais. Methods: This is a quantitative analysis of secondary data based on a cross-sectional research design. Incidence, mortality, date of the first notification of COVID-19 and number of rapid and RT-PCR tests were obtained from the sources: "GAL", "e-SUS VE" and "SES-MG". Pearson coefficient for correlation was calculated to establish the level of association between the relevant data. Descriptive statistical procedures were used to provide a comprehensive understanding of the distribution of incidence, mortality and test rates in the territory. Results: Positive correlations were found between the rate of rapid tests and incidence; rate of RT-PCR tests and incidence/mortality. At the municipal level, incidence, mortality, rate of rapid tests and RT-PCR revealed a negative correlation with days elapsed since the First Notified Case. The same effect occurs at the level of health macro-regions. Conclusion: The heterogeneity of the incidence and mortality of COVID-19 in the territory of Minas Gerais, as well as the rate of tests may be caused, in part, due to the different dates of introduction of the virus in the municipalities/macro-regions. It is speculated that this phenomenon occurs due to the dynamics of regional and inter-regional flows of people.(AU)

Justificativa e Objetivos: Um novo tipo de coronavírus, SARS-CoV-2, é responsável por uma pandemia sem precedentes com profundas consequências socioeconômicas. Devido à sua recente descoberta, o vírus surgido na cidade chinesa de Wuhan, em dezembro de 2019, ainda lança grandes incógnitas. Este estudo objetiva estabelecer as associações espaço-temporais da incidência; mortalidade; e taxas de testes rápidos e RT-PCR em Minas Gerais. Métodos: Trata-se de uma análise quantitativa de dados secundários a partir de um desenho de pesquisa transversal. Incidência, mortalidade, data da(s) primeira(s) notificações da doença, número de testes rápidos e de RT-PCR foram obtidos nas fontes: "Gerenciador de Ambiente Laboratorial", "e-sus VE" e SES-MG. O coeficiente de Pearson para correlação foi calculado para estabelecer o nível de associação entre os dados relevantes. Técnicas estatísticas descritivas foram empregadas para compreender a distribuição da incidência, mortalidade e taxas de testes no território. Resultados: Correlações positivas foram encontradas entre taxa de testes rápidos e incidência; taxa de testes RT-PCR e incidência/mortalidade. A nível municipal, incidência, mortalidade, taxa de testes rápidos e de RT-PCR têm correlação negativa com dias transcorridos desde o Primeiro Caso Notificado. O mesmo efeito ocorre, em diferentes intensidades, a nível das macrorregiões de saúde. Conclusão: A heterogeneidade da incidência e mortalidade da COVID-19 no território mineiro, assim como, das taxas de testes (rápidos e RT-PCR) pode ser causada, em parte, devido às diferentes datas de introdução do vírus nos municípios/macrorregiões de saúde. Especula-se que esse fenômeno se deve às dinâmicas dos fluxos regionais e inter-regionais de pessoas.(AU)

Justificación y Objetivos: El SARS-CoV-2 es responsable por una pandemia sin precedentes con profundas consecuencias socioeconómicas. Debido a su reciente descubrimiento, este vírus representa una gran incógnita para los investigadores. Así, este estudio tiene como objetivo establecer las asociaciones espacio-temporales de la incidencia, la mortalidad y la tasa de pruebas rápidas y RT-PCR en Minas Gerais. Métodos: Trata-se de un análisis cuantitativo de datos secundarios basado en un diseño de investigación transversal. Incidencia, mortalidad, fecha de la primera notificación de COVID-19 y número de pruebas rápidas y RT-PCR se obtuvieron de las fuentes: "GAL", "e-SUS VE" y "SES-MG". Se calculó el coeficiente de correlación de Pearson para establecer el nivel de asociación entre los datos relevantes. Se utilizaron procedimientos estadísticos descriptivos para proporcionar una comprensión integral de la distribución de la incidencia, la mortalidad y las tasas de prueba en el territorio. Resultados: Se encontraron correlaciones positivas entre la tasa de pruebas rápidas y la incidencia; tasa de pruebas de RT-PCR y incidencia/mortalidad. A nivel municipal, la incidencia, mortalidad, tasa de pruebas rápidas y RT-PCR revelaron una correlación negativa con los días transcurridos desde el Primer Caso Notificado. El mismo efecto ocorre a nivel de macrorregiones de salud. Conclusiones: La heterogeneidad de la incidencia y mortalidad de COVID-19 en el territorio de Minas Gerais, así como la tasa de pruebas puede deberse, en parte, a las diferentes fechas de introducción de la virus en los territorios. Se especula que este fenómeno ocurre debido a la dinámica de los flujos regionales e interregionales de personas.(AU)

Peanuts in the air - clinical and experimental studies.

BACKGROUND: Allergic reactions to food allergens usually occur after ingestion. However, fear of reactions to airborne peanut is a common concern for people with peanut allergy. There are no scientific reports on severe reactions with airborne peanut allergen. OBJECTIVE: To investigate the occurrence of allergic reactions in peanut-allergic children undergoing airborne peanut challenge and to determine levels of airborne peanut protein in a separate experimental evaluation. METHODS: Eighty-four children with peanut allergy underwent an airborne peanut challenge, 0.5 m from a bowl of peanuts for 30 min under controlled conditions. In a separate experiment, airborne peanut proteins from roasted and dry-roasted peanuts were collected at varying distances and at varying times with an electret SensAbues filter connected to an air pump. Collected airborne peanut proteins were extracted, dissolved and detected by ELISA. Basophil activation test was used to confirm biological activity. RESULTS: No moderate/severe allergic reactions to airborne peanut allergens were observed. Two children (2%) had mild rhino-conjunctivitis which required no treatment. The IgE-antibodies to peanut or Ara h 2 did not predict a reaction. In the experimental set-up, biological active peanut proteins were detected, in a very low amount, in median 166 ng/ml for dry-roasted and 33 ng/ml for roasted peanuts and decreased dramatically when the collection occurred at a greater distance (0.5-2 m) from the peanut source. Increased exposure time did affect the amount of collected peanut protein at 0 m, and the highest median was obtained after 60 min (p = .012); for time trend p = .0006. CONCLUSIONS AND CLINICAL RELEVANCE: Allergic reactions to airborne peanut proteins are rare and cannot be predicted by high levels of IgE-antibodies to peanut or Ara h 2. Only small amounts of biologically active peanut proteins were detected in the air and seem unlikely to trigger moderate/severe allergic reactions.

Graft survival effect of HLA-A allele matching parathyroid allotransplantation.

Permanent hypoparathyroidism is an endocrine disease that is mostly associated with the disruption of the parathyroid glands during surgery. Allotransplantation is the most promising approach for treatment particularly for its cost-effective and exact curative potential. Herein our aim was to evaluate human leukocyte antigen (HLA)-A allele matching effect on clinical improvement and graft survival after parathyroid transplantation. We performed parathyroid transplantation between ABO/Rh compatible recipient and an unrelated donor who has chronic kidney disease. Preoperative immunological tests include panel reactive antibody, T-flow cytometry crossmatch, B-flow cytometry crossmatch, autoflow cytometry crossmatch, and complement-dependent cytotoxicity crossmatch tests were performed. After histopathological evaluation, half of the resected parathyroid gland cells were isolated and transplanted to the omentum surface by laparoscopy. The transplantation outcome was followed up throughout 382 days. The recipient discharged 2 days after transplantation without any complication. During follow-up, calcium and vitamin D supplementation reduced to a one-third dose; even the intact PTH levels remained low. However, clinical improvement was observed by serum calcium levels. The recipient still continues with low-dose supplementation after 382 days of post-transplantation. Parathyroid cell transplantation to the omental tissue is the most promising option even with only one allele matching for patients with using lifelong high-dose supplementation. Clinical improvements and long-term effect of HLA-A allele matching should be evaluated with more studies and in larger cohorts as well.