IgG and IgM antibody formation to spike and nucleocapsid proteins in COVID-19 characterized by multiplex immunoblot assays.

BACKGROUND: Rapid and simple serological assays for characterizing antibody responses are important in the current COVID-19 pandemic caused by SARS-CoV-2. Multiplex immunoblot (IB) assays termed COVID-19 IB assays were developed for detecting IgG and IgM antibodies to SARS-CoV-2 virus proteins in COVID-19 patients. METHODS: Recombinant nucleocapsid protein and the S1, S2 and receptor binding domain (RBD) of the spike protein of SARS-CoV-2 were used as target antigens in the COVID-19 IBs. Specificity of the IB assay was established with 231 sera from persons with allergy, unrelated viral infections, autoimmune conditions and suspected tick-borne diseases, and 32 goat antisera to human influenza proteins. IgG and IgM COVID-19 IBs assays were performed on 84 sera obtained at different times after a positive RT-qPCR test from 37 COVID-19 patients with mild symptoms. RESULTS: Criteria for determining overall IgG and IgM antibody positivity using the four SARS-CoV-2 proteins were developed by optimizing specificity and sensitivity in the COVID-19 IgG and IgM IB assays. The estimated sensitivities and specificities of the COVID-19 IgG and IgM IBs for IgG and IgM antibodies individually or for either IgG or IgM antibodies meet the US recommendations for laboratory serological diagnostic tests. The proportion of IgM-positive sera from the COVID-19 patients following an RT-qPCR positive test was maximal at 83% before 10 days and decreased to 0% after 100 days, while the proportions of IgG-positive sera tended to plateau between days 11 and 65 at 78-100% and fall to 44% after 100 days. Detection of either IgG or IgM antibodies was better than IgG or IgM alone for assessing seroconversion in COVID-19. Both IgG and IgM antibodies detected RBD less frequently than S1, S2 and N proteins. CONCLUSIONS: The multiplex COVID-19 IB assays offer many advantages for simultaneously evaluating antibody responses to different SARS-CoV-2 proteins in COVID-19 patients.

Dermatological and Genital Manifestations of Lyme Disease Including Morgellons Disease.

Although the erythema migrans (EM) skin rash is traditionally considered a hallmark of Lyme disease, other dermatological manifestations of the tickborne disease are less well known. We describe a 49-year-old woman with erosive genital ulcerations, secondary EM rashes and jagged skin lesions associated with Lyme disease. The skin rashes exhibited fibers characteristic of Morgellons disease. Molecular testing confirmed the presence of Borrelia DNA in both vaginal culture and serum specimens. In further studies on a secondary EM lesion containing filaments, Gömöri trichrome staining revealed the presence of collagen in the filaments, while Dieterle and anti-Borrelia immunostaining revealed intracellular and extracellular Borrelia organisms. Intracellular staining for Borrelia was also observed in lymphocytic infiltrates. Lyme disease may present with a variety of genital lesions and dermatological manifestations including Morgellons disease. Careful evaluation is required to determine the presence of Borrelia organisms associated with these dermopathies.

Lyme Disease: Diversity of Borrelia Species in California and Mexico Detected Using a Novel Immunoblot Assay.

BACKGROUND: With more than 300,000 new cases reported each year in the United States of America (USA), Lyme disease is a major public health concern. Borrelia burgdorferi sensu stricto (Bbss) is considered the primary agent of Lyme disease in North America. However, multiple genetically diverse Borrelia species encompassing the Borrelia burgdorferi sensu lato (Bbsl) complex and the Relapsing Fever Borrelia (RFB) group are capable of causing tickborne disease. We report preliminary results of a serological survey of previously undetected species of Bbsl and RFB in California and Mexico using a novel immunoblot technique. METHODS: Serum samples were tested for seroreactivity to specific species of Bbsl and RFB using an immunoblot method based on recombinant Borrelia membrane proteins, as previously described. A sample was recorded as seropositive if it showed immunoglobulin M (IgM) and/or IgG reactivity with at least two proteins from a specific Borrelia species. RESULTS: The patient cohort consisted of 90 patients residing in California or Mexico who met the clinical case definition of chronic Lyme disease. Immunoblot testing revealed that 42 patients were seropositive for Bbsl (Group 1), while 56 patients were seropositive for RFB (Group 2). Eight patients were seropositive for both Bbsl and RFB species. Group 1 included patients who were seropositive for Bbss (14), B. californiensis (eight), B. spielmanii (10), B. afzelii/B. garinii (10), and mixed infections that included B. mayonii (three). Group 2 included patients who were seropositive for B. hermsii (nine), B. miyamotoi (seven), B. turicatae (nine), and B. turcica (two). In the remaining Group 1 and Group 2 patients, the exact Borrelia species could not be identified using the immunoblot technique. CONCLUSIONS: Lyme disease is associated with a diverse group of Borrelia species in California and Mexico. Current testing for Lyme disease focuses on detection of Bbss, possibly resulting in missed diagnoses and failure to administer appropriate antibiotic therapy in a timely manner. The genetic diversity of Borrelia spirochetes must be considered in future Lyme disease test development.

Line Immunoblot Assay for Tick-Borne Relapsing Fever and Findings in Patient Sera from Australia, Ukraine and the USA.

Tick-borne relapsing fever (TBRF) is caused by spirochete bacteria of the genus Borrelia termed relapsing fever Borreliae (RFB). TBRF shares symptoms with Lyme disease (LD) caused by related Lyme disease Borreliae (LDB). TBRF and LD are transmitted by ticks and occur in overlapping localities worldwide. Serological detection of antibodies used for laboratory confirmation of LD is not established for TBRF. A line immunoblot assay using recombinant proteins from different RFB species, termed TBRF IB, was developed and its diagnostic utility investigated. The TBRF IBs were able to differentiate between antibodies to RFB and LDB and had estimated sensitivity, specificity, and positive and negative predictive values of 70.5%, 99.5%, 97.3%, and 93.4%, respectively, based on results with reference sera from patients known to be positive and negative for TBRF. The use of TBRF IBs and analogous immunoblots for LD to test sera of patients from Australia, Ukraine, and the USA with LD symptoms revealed infection with TBRF alone, LD alone, and both TBRF and LD. Diagnosis by clinical criteria alone can, therefore, underestimate the incidence of TBRF. TBRF IBs will be useful for laboratory confirmation of TBRF and understanding its epidemiology worldwide.

Pilot Study of Immunoblots with Recombinant Borrelia burgdorferi Antigens for Laboratory Diagnosis of Lyme Disease.

Accurate laboratory diagnosis of Lyme disease (Lyme borreliosis), caused by the spirochete Borrelia burgdorferi (BB), is difficult and yet important to prevent serious disease. The US Centers for Disease Control and Prevention (CDC) presently recommends a screening test for serum antibodies followed by confirmation with a more specific Western blot (WB) test to detect IgG and IgM antibodies against antigens in whole cell lysates of BB. Borrelia species related to BB cause tick-borne relapsing fever (TBRF). TBRF is increasingly recognized as a health problem in the US and occurs in areas where Lyme disease is prevalent. The two groups of Borrelia share related antigens. We have developed a modified WB procedure termed the Lyme immunoblots (IBs) using recombinant antigens from common strains and species of the BB sensu lato complex for serological diagnosis of Lyme disease. A reference collection of 178 sera from 26 with and 152 patients without Lyme disease were assessed by WB and IB in a blinded manner using either criteria for positive antibody reactions recommended by the CDC or criteria developed in-house. The sensitivity, specificity, positive and negative predictive values obtained with the reference sera suggest that the Lyme IB is superior to the Lyme WB for detection of specific antibodies in Lyme disease. The Lyme IB showed no significant reaction with rabbit antisera produced against two Borrelia species causing TBRF in the US, suggesting that the Lyme IB may be also useful for excluding TBRF.

Detection of tick-borne infection in Morgellons disease patients by serological and molecular techniques.

BACKGROUND: Morgellons disease (MD) is a skin condition associated with Lyme disease (LD) and tick-borne illness. Patients with this skin disorder experience ulcerative lesions that contain multicolored filamentous collagen and keratin inclusions. Infection with various species of Borrelia and other tick-borne pathogens has been detected in tissue and body fluid specimens from MD patients. We sought to explore this association further in a cohort of MD patients. PATIENTS AND METHODS: Sera from 30 patients with MD were tested for antibody reactivity to antigens from the Borrelia burgdorferi (Bb) group and the relapsing fever Borrelia (RFB) group of spirochetes. Tissue and/or body fluid specimens from these patients were also tested for the presence of Bb and RFB infection using PCR technology. In addition, tissue and body fluid specimens were tested for the presence of Bartonella henselae using PCR, and formalin-fixed skin sections from a subset of patients were tested using fluorescent in situ hybridization (FISH) with B. henselae-specific DNA probes. RESULTS: Seroreactivity to Bb, RFB or both was detected in 63% of the cohort, while positive PCR testing for Bb, RFB or both was detected in 53% of the cohort. Overall, 90% of patients tested positive for exposure and/or infection with Borrelia spirochetes. B. henselae infection was detected by PCR in skin sections or body fluids from 20% of the subjects, and B. henselae FISH testing was positive in 30% of the dermatological specimens submitted for study. CONCLUSION: The study demonstrates an association between MD and positive tests for both Bb and RFB spirochetes. In conjunction with previous studies, our study provides corroborative evidence linking MD to Borrelia infection and tick-borne illness.

High yield selective acylation of polyamines: proton as protecting group.

An advance in the selective acylation of polyamines having identical or similar amine functions is reported. While nucleophilicity differences between the various amine functions are slight, the corresponding conjugate acids exhibit pKa values over a significant range. We have used proton as polyamine protecting group: the monoamine resulting from single deprotonation of a polyammonium compound has allowed for high yields of selective acylation.

Asociación entre salud mental y actitudes ante el confinamiento durante la pandemia COVID-19 en hospitales de Lima y Callao

Objetivo : Identificar la asociación entre la depresión, ansiedad y estrés con las actitudes ante el confinamiento durante la pandemia por COVID-19 en personal de salud del Hospital Nacional Daniel Alcides Carrión y Hospital Militar Central mediante el empleo de la escala DASS-21. Materiales y métodos : Estudio transversal, observacional y analítico, participaron personal de salud que trabajó en áreas Covid durante la pandemia COVID-19. Se empleó la escala Dass-21 para medir la existencia de estados emocionales de depresión, ansiedad y estrés, y la escala de actitudes adaptada frente al confinamiento para COVID-19. Resultados : Se incluyó 124 participantes, 54% (n=67) de sexo femenino, con una mediana de edad de 34.5 años [Ri]= 16.5 que reportaron una mediana de años de experiencia de 6 (Ri = 17). La mediana del puntaje de la Escala Dass-21 para ansiedad fue de 2 (Ri= 5), la mediana del puntaje de Dass-21 para depresión fue de 1 (Ri=4.5) y la mediana del puntaje de la Escala Dass-21 para estrés fue de 4 (Ri=5). El análisis bivariado entre los valores de los parámetros de la Escala Dass-21 y las demás covariables identificó que el grado de instrucción tenía una relación significativa con la subescala de depresión, ansiedad y estrés. Conclusión : Se encontró asociación entre los parámetros afectivos de la escala sobre actitudes ante el confinamiento y las tres subescalas de DASS-21.

Objective : Identify the association between depression, anxiety and stress with attitudes towards confinement during the COVID-19 pandemic in health staff from the Daniel Alcides Carrion National Hospital and Central Military Hospital by using the DASS-21 scale. Materials and methods : Cross-sectional, observational and analytical study, health staff participated who worked in COVID areas during the Covid-19 pandemic. The Dass-21 scale was used to measure the existence of emotional states of depression, anxiety and stress, and the scale of attitudes adapted to confinement for Covid-19. Results : 124 participants were included, 54% female (n=67), with a median age of 34.5 years [Ri]= 16.5, who reported a median year of experience of 6 (Ri = 17). The median Dass-21 score for anxiety was 2 (Ri = 5), the median Dass-21 score for depression was 1 (Ri = 4.5), and the median Dass-21 scale score for stress was 4 (Ri = 5). The bivariate analysis between the values of the Dass-21 Scale domains and the other covariates identified that the level of education had a highly significant relationship with the depression subscale, anxiety and stress. Conclusion : An association was found between the affective parameters of the scale on attitudes towards confinement and the three subscales of DASS-21.

Lowering risk for early alcohol use by challenging alcohol expectancies in elementary school children.

Altering alcohol expectancies has reduced alcohol use among young adults and may lead to successful prevention of early alcohol use. The authors randomly assigned 216 4th-grade children to an expectancy challenge or control condition and used individual-differences scaling to map expectancies into memory network format, with preference mapping to model likely paths of association. After expectancy intervention, children exhibited a greater likelihood to associate alcohol use with negative and sedating consequences and a decreased likelihood to associate alcohol with positive and arousing consequences. Children and adults who emphasize negative and sedating effects have been found to be less likely touse alcohol. Therefore, expectancy challenge interventions that have been successful at modifying expectancies and subsequently decreasing alcohol consumption of adults may be useful in reducing the likelihood of early alcohol use among children.

Evaluación de la paciente con incontinencia urinaria / Evaluation of patients with urinary incontinence

El clínico debe conocer bien la terminología empleada universalmente de modo de captar adecuadamente los síntomas y etiquetarlos según la etiología de la incontinencia urinaria. Dos son las causas de incontinencia más frecuentes y corresponden a las originadas por el esfuerzo físico. La tendencia creciente es considerar que las pruebas urodinámicas, particularmente el cistometro grama es el único método que permite establecer la diferenciación de ambas formas, con exactitud. Se enfatiza la necesidad de contar con un cuestionario y protocolo de estudio apropiados que permitan captar los síntomas básicos y orientar su estudio. Se ha popularizado el empleo de la prueba del hisopo de algodón y es de utilización creciente la del peso de la toalla sanitaria en la evaluación de la paciente incontinente. Los estudios radiológicos juegan un papel importante en la investigación de las causas de incontinencia. La uretroscopia es un recurso útil en a diferención de algunas formas de incontinencia, pero indudablemente lo más importante es el adecuado juicio clínico del médico tratante

La prevalencia del síndrome de antifosfolípidos en pacientes con lupus eritematoso sistémico en el Hospital Universitario de Puerto Rico / Prevalence of antiphospholipid syndrome in patients with systemic lupus erythematosus at the University Hospital of Puerto Rico

This is a retrospective study based on a population of 80 patients with connective tissue diseases from the University Hospital of Puerto Rico. Among the population, 62 (77.5) of the patients had Systemic Lupus Erythematosus (SLE), whom we were most interested to monitor. The investigation revealed an incidence of 13.8 of anthiophospholipid syndrone within the general population. Among the patients with SLE it was 12.9, and only 5.5 among the population with other diagnoses. The antibody found with the highest frequency within the systemic lupus erythematosus population was the anticardiolipin IgG (30.6) and lupus anticoagulant (17.7). The antibody frequency among patients with other diagnoses was only 5.5 for lupus anticoagulant and 5.5 for anticardiolipin IgM (the only one found). Among SLE's clinical manifestations, the most frequently found were thrombocitopenia and pregnancy complications.