Development and Validation of a Highly Sensitive Multiplex Immunoassay for SARS-CoV-2 Humoral Response Monitorization: A Study of the Antibody Response in COVID-19 Patients with Different Clinical Profiles during the First and Second Waves in Cadiz, Spain.

There is still a long way ahead regarding the COVID-19 pandemic, since emerging waves remain a daunting challenge to the healthcare system. For this reason, the development of new preventive tools and therapeutic strategies to deal with the disease have been necessary, among which serological assays have played a key role in the control of COVID-19 outbreaks and vaccine development. Here, we have developed and evaluated an immunoassay capable of simultaneously detecting multiple IgG antibodies against different SARS-CoV-2 antigens through the use of Bio-PlexTM technology. Additionally, we have analyzed the antibody response in COVID-19 patients with different clinical profiles in Cadiz, Spain. The multiplex immunoassay presented is a high-throughput and robust immune response monitoring tool capable of concurrently detecting anti-S1, anti-NC and anti-RBD IgG antibodies in serum with a very high sensitivity (94.34-97.96%) and specificity (91.84-100%). Therefore, the immunoassay proposed herein may be a useful monitoring tool for individual humoral immunity against SARS-CoV-2, as well as for epidemiological surveillance. In addition, we show the values of antibodies against multiple SARS-CoV-2 antigens and their correlation with the different clinical profiles of unvaccinated COVID-19 patients in Cadiz, Spain, during the first and second waves of the pandemic.

From Cell to Symptoms: The Role of SARS-CoV-2 Cytopathic Effects in the Pathogenesis of COVID-19 and Long COVID.

Severe Acute Respiratory Syndrome CoronaVirus 2 (SARS-CoV-2) infection triggers various events from molecular to tissue level, which in turn is given by the intrinsic characteristics of each patient. Given the molecular diversity characteristic of each cellular phenotype, the possible cytopathic, tissue and clinical effects are difficult to predict, which determines the heterogeneity of COVID-19 symptoms. The purpose of this article is to provide a comprehensive review of the cytopathic effects of SARS-CoV-2 on various cell types, focusing on the development of COVID-19, which in turn may lead, in some patients, to a persistence of symptoms after recovery from the disease, a condition known as long COVID. We describe the molecular mechanisms underlying virus-host interactions, including alterations in protein expression, intracellular signaling pathways, and immune responses. In particular, the article highlights the potential impact of these cytopathies on cellular function and clinical outcomes, such as immune dysregulation, neuropsychiatric disorders, and organ damage. The article concludes by discussing future directions for research and implications for the management and treatment of COVID-19 and long COVID.

Specific Activation of T Cells by an ACE2-Based CAR-Like Receptor upon Recognition of SARS-CoV-2 Spike Protein.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the causative agent of the Coronavirus Disease 2019 (COVID-19) pandemic, which is still a health issue worldwide mostly due to a high rate of contagiousness conferred by the high-affinity binding between cell viral receptors, Angiotensin-Converting Enzyme 2 (ACE2) and SARS-CoV-2 Spike protein. Therapies have been developed that rely on the use of antibodies or the induction of their production (vaccination), but despite vaccination being still largely protective, the efficacy of antibody-based therapies wanes with the advent of new viral variants. Chimeric Antigen Receptor (CAR) therapy has shown promise for tumors and has also been proposed for COVID-19 treatment, but as recognition of CARs still relies on antibody-derived sequences, they will still be hampered by the high evasion capacity of the virus. In this manuscript, we show the results from CAR-like constructs with a recognition domain based on the ACE2 viral receptor, whose ability to bind the virus will not wane, as Spike/ACE2 interaction is pivotal for viral entry. Moreover, we have developed a CAR construct based on an affinity-optimized ACE2 and showed that both wild-type and affinity-optimized ACE2 CARs drive activation of a T cell line in response to SARS-CoV-2 Spike protein expressed on a pulmonary cell line. Our work sets the stage for the development of CAR-like constructs against infectious agents that would not be affected by viral escape mutations and could be developed as soon as the receptor is identified.

How Has the Aspergillosis Case Fatality Rate Changed over the Last Two Decades in Spain?

(1) Background: Aspergillus produces high morbidity and mortality, especially in at-risk populations. In Spain, the evolution of mortality in recent years due to this fungus is not well established. The aim of this study was to estimate the case fatality rate of aspergillosis in inpatients from 1997 to 2017 in Spain. (2) Methodology: A retrospective descriptive study was conducted with records of inpatients admitted to the National Health System with a diagnosis of aspergillosis. (3) Principal findings: Of 32,960 aspergillosis inpatients, 24.5% of deaths were registered, and 71% of the patients who died were men. The percentage of deaths increased progressively with age. The case fatality rate progressively decreased over the period, from 25.4 and 27.8% in 1997-1998 to values of 20.6 and 20.8% in 2016 and 2017. Influenza and pneumonia occurrence/association significantly increased case fatality rates in all cases. (4) Conclusions: Our study shows that lethality significantly decreased in the last two decades despite the increase in cases. This highlights the fact that patients with solid and/or hematological cancer do not have a much higher mortality rate than the group of patients with pneumonia or influenza alone, these two factors being the ones that cause the highest CFRs. We also need studies that analyze the causes of mortality to decrease it and studies that evaluate the impact of COVID-19.

Solubilización in vitro del aceite de árbol de té y primeros resultados de su efecto antifúngico en onicomicosis / Solubilization in vitro of tea tree oil and first results of antifungal effect in onychomycosis

Introducción: La onicomicosis por dermatofitos son la principal causa de alteración ungueal. Su abordaje terapéutico farmacológico tiene asociado bajas tasas de éxito, hepatotoxicidad, interferencia y baja adherencia al tratamiento. Por ello, nuestro objetivo fue valorar la efectividad in vitro del aceite esencial de árbol de té, para aportar alternativas menos nocivas en el abordaje terapéutico frente a los principales agentes causantes de estas infecciones. Material y métodos: Se aisló de fragmentos de uña con infección fúngica el Trichophyton rubrum y Trichophyton mentagrophytes. Posteriormente, se inocularon a una concentración de 3 x105 UFC/mL en agar patata con dextrosa y aceite esencial de árbol de té a diferentes concentraciones para evaluar su efecto mediante el recuento de unidades formadoras de colonia (UFC) y el crecimiento radial (CR). Resultados: Se obtuvo compromiso en el crecimiento de Trichophyton rubrum a concentraciones a partir de 0,04% del aceite esencial de árbol de té (p = 0,004). En el caso del Trichophyton mentagrophytes se obtuvo inhibición a partir de 0,02% (p = 0,017), e incluso la inhibición completa a una concentración final del aceite a 0,07%. Conclusiones: El aceite esencial de árbol de té inhibe el crecimiento in vitro de los hongos estudiados, pudiéndose considerar una alternativa menos nociva para el abordaje terapéutico de las onicomicosis.(AU)

Introduction: Onychomycosis is the main cause of nail alteration. Hepatotoxicity, interference and low adherence to pharmacological treatment are associated. Therefore, our objective was to assess the in vitro effectiveness of tea tree essential oil (less harmful) against main causative agents of these infections. Material and methods: Trichophyton rubrum and Trichophyton mentagrophytes were isolated and inoculated at a concentration of 3x105 CFU / mL in potato agar dextrose and tea tree essential oil at different concentrations to assess its effect by counting colony forming units and radial growth. Results: Trichophyton rubrum growth inhibition was obtained at concentrations higher than 0.04% of the essential tea tree oil (p = 0.004). In the case of Trichophyton mentagrophytes, inhibition was obtained at 0.02% (p = 0.017), and even complete inhibition at a final concentration of the oil at 0.07%. Conclusions: Tea tree essential oil inhibits the in vitro growth of the fungus and may be a less harmful alternative to the onychomycosis treatment.(AU)

Solubilization in vitro of tea tree oil and first results of antifungal effect in onychomycosis.

INTRODUCTION: Onychomycosis is the main cause of nail alteration. Hepatotoxicity, interference and low adherence to pharmacological treatment are associated. Therefore, our objective was to assess the in vitro effectiveness of tea tree essential oil (less harmful) against main causative agents of these infections. MATERIAL AND METHODS: Thrichophyton rubrum and Thrichophyton mentagrophytes were isolated and inoculated at a concentration of 3 × 105 CFU/mL in potato agar dextrose and tea tree essential oil at different concentrations to assess its effect by counting colony forming units and radial growth. RESULTS: Trichophyton rubrum growth inhibition was obtained at concentrations higher than 0.04% of the essential tea tree oil (p = 0.004). In the case of Thrichophyton mentagrophytes, inhibition was obtained at 0.02% (p = 0.017), and even complete inhibition at a final concentration of the oil at 0.07%. CONCLUSIONS: Tea tree essential oil inhibits the in vitro growth of the fungus and may be a less harmful alternative to the onychomycosis treatment.

Epidemiology of aspergillosis in hospitalised Spanish patients-A 21-year retrospective study.

BACKGROUND: Aspergillosis is a serious infection, and in Spain, the influence of epidemiology and climate on the resulting expenses of aspergillosis is not well established. AIM: A retrospective descriptive study using the Minimum Basic Data Set was performed on records of patients admitted to hospitals of the National Health System between 1997 and 2017 with a diagnosis of aspergillosis. The weather parameters were obtained from the State Agency of Meteorology from Spain. RESULTS: A total of 32,960 patients were identified, of whom 22,383 were men (68%). The mean age (±SD) was 61.1 ± 19.1 years. The mean incidence rate for all diagnoses was 3.54 cases per 100,000 person-years (95% CI, 3.50-3.57). The incidence rate in men was twice as high as that in women, 4.89 (95% CI, 4.82-4.95) vs. 2.24 (95% CI, 2.19-2.27) cases per 100,000 person-years (p > .001). The highest incidence rates were concentrated in northern Spain. One in four patients died (8,080 cases; 25%). There was a moderate positive linear association between rainfall and incidence rate (rP  = .508; p = .026). In contrast, the Pearson's correlation coefficient indicated a moderate negative linear relationship between temperature and incidence rate (rP  = -.447; p = .050). We observed a higher incidence in the months with higher humidity and rainfall. CONCLUSIONS: Our study supports a high burden of aspergillosis in Spain, with an increase in cases in the past two decades. Additionally, the influence of climatological factors on the incidence of aspergillosis is highlighted. Despite preemptive treatment strategies, this infection still has a high mortality.

Repetición de las pruebas microbiológicas en la sospecha de la infección por SARS-CoV-2: utilidad de un score basado en la probabilidad clínica / Repetition of microbiological tests in suspect of SARS-CoV-2 infection: utility of a score based on clinical probability

INTRODUCCIÓN: El diagnóstico de la infección por SARSCoV-2 presenta limitaciones. La RT-PCR en frotis nasofaríngeo es la prueba considerada como el patrón oro, aunque puede ofrecer falsos negativos. El objetivo de este trabajo ha sido evaluar la utilidad de repetir el frotis nasofaríngeo en pacientes con resultado negativo en función de las diferentes probabilidades clínicas. MÉTODOS: Estudio observacional retrospectivo de los primeros pacientes ingresados en el Hospital Universitario Marqués de Valdecilla en dos plantas COVID de Medicina Interna durante marzo-abril del 2020. La RT-PCR para la detección de al menos dos dianas de los principales genes (E, N, RdRP, ORFab1) y el test de detección de anticuerpos para la detección de al menos IgG. RESULTADOS: Se analizaron 145 pacientes hospitalizados; 98 (67,5%) con el diagnóstico de SARS-CoV-2. Las variables predictivas independientes de SARS-CoV-2 fueron: contacto epidemiológico con otro paciente COVID-19, presentación clínica como neumonía, ausencia de neumonía en el año previo, inicio de síntomas >7 días al ingreso, ≥2 síntomas -tos, disnea y fiebre- y lactato deshidrogenasa >350 U/L (p < 0,05). Un score basado en estas variables presentó un área bajo la curva ROC (ABC ROC) de 0,89 (IC95 0,831-0,946; p < 0,001). El rendimiento de la RT-PCR en su primera determinación fue del 54,9%. La repetición de la prueba permitió detectar un 16% de casos adicionales. El rendimiento global de sucesivas RT-PCR en pacientes con baja probabilidad clínica fue inferior al 5%. CONCLUSIÓN: Hemos definido un score de probabilidad pre-prueba basado en datos epidemiológicos y clínicos con una buena precisión para el diagnóstico por infección SARSCoV-2. La repetición del frotis nasofaríngeo puede evitar errores de muestreo sólo en escenarios de intermedia-alta probabilidad clínica pre-prueba

BACKGROUND: The diagnosis of SARS-CoV-2 infection presents some limitations. RT-PCR in nasopharyngeal swabs is considered the gold standard for the diagnosis, although it can have false negative results. We aimed to analyze the accuracy of repeating nasopharyngeal swabs based on different clinical probabilities. METHODS: Retrospective observational study of the first patients admitted to a two COVID Internal Medicine wards at the University Hospital Marqués de Valdecilla, Santander, from March to April 2020. RT-PCR targering E, N, RdRP and ORFab1 genes and antibody tests detecting IgG. RESULTS: A total of 145 hospitalized patients with suspected SARS-Cov2 infection were admitted and in 98 (67.5%) diagnosis was confirmed. The independent predictive variables for SARS-CoV-2 infection were: epidemiological contact, clinical presentation as pneumonia, absence of pneumonia in the last year, onset of symptoms > 7 days, two or more of the following symptoms -dyspnea, cough or fever- and serum lactate dehydrogenase levels >350 U/L (p < 0.05). A score based on these variables yielded an AUC-ROC of 0.89 (CI95%, 0.831-0.946; p < 0.001). The accuracy of the first nasopharyngeal swabs was 54.9%. Repeating nasopharyngeal swabs two or three times allows to detect an additional 16% of positive cases. The overall accuracy of successive RT-PCR tests in patients with low pre-test probability was <5%. CONCLUSIONS: We have defined a pre-test probability score based on epidemiological and clinical data with a high accuracy for diagnosis of SARS-CoV-2. Repeating nasopharyngeal swabs avoids sampling errors, but only in medium of high probability pre-test clinical scenarios

Solubilization in vitro of tea tree oil and first results of antifungal effect in onychomycosis. / Solubilización in vitro del aceite de árbol de té y primeros resultados de su efecto antifúngico en onicomicosis.

INTRODUCTION: Onychomycosis is the main cause of nail alteration. Hepatotoxicity, interference and low adherence to pharmacological treatment are associated. Therefore, our objective was to assess the in vitro effectiveness of tea tree essential oil (less harmful) against main causative agents of these infections. MATERIAL AND METHODS: Trichophyton rubrum and Trichophyton mentagrophytes were isolated and inoculated at a concentration of 3x105 CFU / mL in potato agar dextrose and tea tree essential oil at different concentrations to assess its effect by counting colony forming units and radial growth. RESULTS: Trichophyton rubrum growth inhibition was obtained at concentrations higher than 0.04% of the essential tea tree oil (p = 0.004). In the case of Trichophyton mentagrophytes, inhibition was obtained at 0.02% (p = 0.017), and even complete inhibition at a final concentration of the oil at 0.07%. CONCLUSIONS: Tea tree essential oil inhibits the in vitro growth of the fungus and may be a less harmful alternative to the onychomycosis treatment.

I Can Not Wear My Sunglasses: An Unusual Sarcoidosis Presentation.

A 39-year-old Caucasian man was referred to University Hospital Salamanca from a primary care unit due to the presence of an erythematous violaceous nodule at the superior portion of his nose. Physical examination indicated that the firm, fixed erythematous violaceous nodule measured approximately 2 cm in diameter and was located inferior to a scar on the nasal bridge. Cutaneous involvement in sarcoidosis occurs in 25% of cases. A wide range of clinical presentations of cutaneous sarcoidosis is recognized. Skin lesions are classified as either non-specific, of which erythema nodosum is the most representative and specific, or as granulomatous, which includes maculopapular nodules, plaques, infiltrated scars, lupus pernio, ulcerations, warty lesions and erythroderma. Scar sarcoidosis is a type of cutaneous sarcoidosis.

Efectividad de una intervención educativa breve en pacientes con insomnio en atención primaria / Effectiveness of short educational intervention in patients with insomnia in primary care

OBJETIVO: Evaluar la efectividad de asociar una intervención educativa breve al tratamiento farmacológico para el insomnio en atención primaria (AP). DISEÑO: Ensayo clínico no aleatorizado simple ciego. Emplazamiento: Dos centros de salud urbanos en Gijón (Asturias). Participantes: Pacientes que consultaron por insomnio entre julio de 2012 y enero de 2013 y cumplieron criterios de inclusión. Fueron asignados sistemáticamente al grupo intervención (GI) o al grupo control (GC) tras aleatorizar al primer paciente de cada grupo. Todos iniciaron lorazepam 1 mg nocturno y acudieron a 4 visitas de 15 min y una visita de revisión al mes siguiente. Intervención: El GI fue instruido en medidas de control de estímulos, higiene del sueño y técnicas de relajación y respiración. Al GC se le hicieron mediciones no invasivas. Mediciones principales: Se consideró curados a quienes obtuvieron Índice de Calidad de Sueño de Pittsburg (ICSP) < 6 o reducción del 50%. Se analizó secundariamente el descenso del ICSP tras las visitas (visita final) y al mes siguiente (visita revisión), y consumo de lorazepam en la visita revisión. Análisis: inferencia bayesiana. RESULTADOS: Se obtuvieron 12 curaciones entre 26 intervenidos, y una entre 24 controles. Las medias posteriores del descenso de ICSP en visita final y revisión fueron de: -4,7 (ICred95%: -5,9 a -3,5) y -6,3 (ICred95%: -7,5 a -5,1) en GI, frente a -1,8 (ICred95%: -3 a -0,5) y -1,7 (ICred95%: -2,9 a -0,4) en GC. Abandonaron lorazepam 9 intervenidos (34,6%) frente a 4 controles (16,7%). Completaron el estudio 19 intervenidos y 17 controles. El análisis por protocolo no mostró diferencias relevantes. CONCLUSIONES: Implementar una intervención educativa breve en AP mejora la calidad del sueño y potencia una menor prescripción de benzodiacepinas

OBJECTIVE: To evaluate the effectiveness of providing an educational intervention in primary care (PC) alongside a pharmacological treatment for insomnia. DESIGN: Single blinded non randomised clinical trial. Location: Two urban primary health centers in Gijón (Asturias, Spain). Participants: Patients who consulted for insomnia between July 2012-January 2013 and met the inclusion criteria (n = 50) were assigned systematically to the control group (CG) or intervention group (IG). All patients initiated treatment with lorazepam 1 mg in the evenings and had four weekly 15 min visits plus a follow-up visit after another month. Interventions: The IG received training for control of stimuli, sleep hygiene and respiration and relaxation techniques in the four visits. The CG had only non invasive measures taken. Measurements: Considering as cured those who reached a Pittsburgh Sleep Quality Index PSQI < 6 or a 50% reduction from baseline level. It was also analyzed the change in the PSQI from baseline to final visit and to follow-up visit, and voluntary interruption of lorazepam. Analysis by Bayesian inference. RESULTS: Twelve out of recoveries after intervention against one out of 24 among control group. Mean change in PSQI to final visit and follow-up visit was: -4.7 (95%CrI: -5.9 to -3.5) and -6,3 (95%ICred: -7.5 to -5.1) in IG; -1.8 (95%ICred: -3 to -0.5) and -1.7 (95%ICred: -2.9 to -0.4) in CG. Interruption of lorazepan: in 4 controls (16,7%) and 9 (34,6%) in IG. Twenty nine patients in GI and 17 in GC completed the trial. Per protocol analysis showed similar results. CONCLUSIONS: The educational intervention in PC improves sleep quality and reduces the need of treatment with benzodiacepines

[Effectiveness of short educational intervention in patients with insomnia in primary care]. / Efectividad de una intervención educativa breve en pacientes con insomnio en atención primaria.

OBJETIVE: To evaluate the effectiveness of providing an educational intervention in primary care (PC) alongside a pharmacological treatment for insomnia. DESIGN: Single blinded non randomised clinical trial. LOCATION: Two urban primary health centers in Gijón (Asturias, Spain). PARTICIPANTS: Patients who consulted for insomnia between July 2012-January 2013 and met the inclusion criteria (n=50) were assigned systematically to the control group (CG) or intervention group (IG). All patients initiated treatment with lorazepam 1mg in the evenings and had four weekly 15 min visits plus a follow-up visit after another month. INTERVENTIONS: The IG received training for control of stimuli, sleep hygiene and respiration and relaxation techniques in the four visits. The CG had only non invasive measures taken. MEASUREMENTS: Considering as cured those who reached a Pittsburgh Sleep Quality Index PSQI <6 or a 50% reduction from baseline level. It was also analyzed the change in the PSQI from baseline to final visit and to follow-up visit, and voluntary interruption of lorazepam. Analysis by Bayesian inference. RESULTS: Twelve out of recoveries after intervention against one out of 24 among control group. Mean change in PSQI to final visit and follow-up visit was: -4.7 (95%CrI:-5.9 to -3.5) and -6,3 (95%ICred: -7.5 to -5.1) in IG; -1.8 (95%ICred: -3 to -0.5) and -1.7 (95%ICred: -2.9 to -0.4) in CG. Interruption of lorazepan: in 4 controls (16,7%) and 9 (34,6%) in IG. Twenty nine patients in GI and 17 in GC completed the trial. Per protocol analysis showed similar results. CONCLUSIONS: The educational intervention in PC improves sleep quality and reduces the need of treatment with benzodiacepines.