Cardiovascular Risk Scales Association with Cerebrospinal Fluid Alzheimer's Disease Biomarkers in Cardiovascular Low Cardiovascular Risk Regions.

BACKGROUND: Cardiovascular risk factors are associated with Alzheimer's Disease (AD) development. However, few studies compare the overall cardiovascular risk with AD biomarkers, and when done, they are mainly performed in moderate cardiovascular risk regions. OBJECTIVES: To determine whether cardiovascular risk in older adults is associated with pathological cerebrospinal fluid (CSF) biomarkers of AD in a low cardiovascular risk population. DESIGN: This is a cross-sectional study performed between 2017 and 2020. PARTICIPANTS: The present work included patients between 50 and 75 years old who were negative for CSF AD biomarkers and had minimum cognitive alterations (controls) and patients with positive CSF AD biomarkers and in early stages of AD (cases). MEASUREMENTS: CSF biomarkers included total tau, phosphorylated tau 181 and amyloid ß42 (Aß42). Analytical variables were obtained. ERICE, SCORE2 and Framingham scales were used to calculate the overall patient's cardiovascular risk. The Aß42/Aß40 ratio and neurofilaments were explored when available. RESULTS: Two hundred and thirty-three patients were included. Nearly 76% of the sample had AD. AD patients had higher cardiovascular risk than controls (p-value < 0.05). ERICE and SCORE2 were associated with AD presence. Framingham was not. A correlation between elevated cardiovascular risk and higher total tau and NfL levels was observed when adjusted by age. CONCLUSION: Cardiovascular risk assessment may be helpful in neurodegenerative disorders detection, as it is associated with CSF total tau and NfL. ERICE and SCORE2 may be useful scales in low cardiovascular risk regions to improve cardiovascular control and prevent neurodegenerative pathologies.

Photoanthropometric craniofacial parameters in individuals with osteogenesis imperfecta.

BACKGROUND: This study aimed to evaluate facial photoanthropometric parameters in patients with OI. MATERIAL AND METHODS: We selected 20 Brazilian patients diagnosed with OI treated at the Extension Service for Minors in Need of Specialized Treatment of the Dentistry Course at the Federal University of Ceará (Fortaleza, Brazil), of both sexes, without age restriction, and able to understand and sign the informed consent form (ICF). As a control group, 38 non-syndromic Brazilian individuals, categorized as ASA I, able to understand and sign the ICF, matched by sex, age, and Legan and Burstone facial profile were selected. The exclusion criteria were: previous orthodontic treatment, craniofacial trauma and/or surgery, and the presence of any other systemic diseases. Photoanthropometric analysis of the 18 facial parameters proposed by Stengel-Rutkowski et al. (1984), previously established in the literature for craniofacial syndromes, were conducted. A single examiner digitally performed all effective and angular measurements with the CorelDRAWX7® software. RESULTS: Horizontally shortened ears (p<0.001) but larger in height in relation to the face (p=0.012) were shown to be alterations belonging to individuals with OI. CONCLUSIONS: OI patients present distinct photoanthropometric parameters inherent in this condition.

The effects of hypothetical behavioral interventions on the 13-year incidence of overweight/obesity in children and adolescents.

BACKGROUND: In view of the high burden of childhood overweight/obesity (OW/OB), it is important to identify targets for interventions that may have the greatest effects on preventing OW/OB in early life. Using methods of causal inference, we studied the effects of sustained behavioral interventions on the long-term risk of developing OW/OB based on a large European cohort. METHODS: Our sample comprised 10 877 children aged 2 to < 10 years at baseline who participated in the well-phenotyped IDEFICS/I.Family cohort. Children were followed from 2007/08 to 2020/21. Applying the parametric g-formula, the 13-year risk of developing OW/OB was estimated under various sustained hypothetical interventions on physical activity, screen time, dietary intake and sleep duration. Interventions imposing adherence to recommendations (e.g. maximum 2 h/day screen time) as well as interventions 'shifting' the behavior by a specified amount (e.g. decreasing screen time by 30 min/day) were compared to 'no intervention' (i.e. maintaining the usual or so-called natural behavior). Separately, the effectiveness of these interventions in vulnerable groups was assessed. RESULTS: The 13-year risk of developing OW/OB was 30.7% under no intervention and 25.4% when multiple interventions were imposed jointly. Meeting screen time and moderate-to-vigorous physical activity (MVPA) recommendations were found to be most effective, reducing the incidence of OW/OB by -2.2 [-4.4;-0.7] and -2.1 [-3.7;-0.8] percentage points (risk difference [95% confidence interval]), respectively. Meeting sleep recommendations (-0.6 [-1.1;-0.3]) had a similar effect as increasing sleep duration by 30 min/day (-0.6 [-0.9;-0.3]). The most effective intervention in children of parents with low/medium educational level was being member in a sports club; for children of mothers with OW/OB, meeting screen time recommendations and membership in a sports club had the largest effects. CONCLUSIONS: While the effects of single behavioral interventions sustained over 13 years were rather small, a joint intervention on multiple behaviors resulted in a relative reduction of the 13-year OW/OB risk by between 10 to 26%. Individually, meeting MVPA and screen time recommendations were most effective. Nevertheless, even under the joint intervention the absolute OW/OB risk remained at a high level of 25.4% suggesting that further strategies to better prevent OW/OB are required.

Level sets of depth measures in abstract spaces.

The lens depth of a point has been recently extended to general metric spaces, which is not the case for most depths. It is defined as the probability of being included in the intersection of two random balls centred at two random points X and Y, with the same radius d(X, Y). We prove that, on a separable and complete metric space, the level sets of the empirical lens depth based on an iid sample, converge in the Painlevé-Kuratowski sense, to its population counterpart. We also prove that, restricted to compact sets, the empirical level sets and their boundaries are consistent estimators, in Hausdorff distance, of their population counterparts, and analyse two real-life examples.

Estudio de los factores de riesgo de deterioro cognitivo en el medio rural: metodología y pilotaje desde la farmacia comunitaria / Assessing risk factors for cognitive impairment in rural areas: methodology and pilot study in community pharmacy

Introducción: el deterioro cognitivo está mediado por la exposición a una gran variedad de factores de riesgo, entre los que la edad es el más relevante. El fenómeno global del envejecimiento de la población hace que conseguir separar los conceptos de envejecimiento y enfermedad sea una tarea de gran interés. En España, la población rural es un estrato particularmente envejecido que puede tener dificultades de acceso a los servicios sanitarios. Las farmacias comunitarias están ampliamente distribuidas en nuestra geografía y el farmacéutico es el sanitario más accesible para esta población tan vulnerable. Métodos: se ha diseñado y pilotado una metodología para evaluar el deterioro cognitivo y el envejecimiento saludable desde la farmacia comunitaria en nuestro medio, utilizando un cuestionario ad hoc compuesto por tests validados para población española, dividido en cinco secciones: deterioro cognitivo, factores sociodemográficos, problemas de salud, estilo de vida y factores psicosociales. Resultados: la prevalencia encontrada para el deterioro cognitivo y el envejecimiento saludable fue de 28,2 % y 15,4 %, respectivamente. La prevalencia (o el valor medio poblacional) calculado para factores de riesgo o factores protectores de deterioro cognitivo resultó compatible con los datos publicados para población española. Los resultados mostraron que este cuestionario puede resultar adecuado para la recogida de información acerca de las variables relacionadas con el deterioro cognitivo en nuestro entorno. Conclusión: el estudio de la influencia de cada factor sobre el deterioro cognitivo debe ser abordado con detalle debido a las complejas interrelaciones existentes entre ellos. En este sentido, las variables más novedosas, como son los factores psicosociales, deben constituir uno de los objetivos prioritarios (AU)

Composición corporal en mujeres deportistas premenopáusicas y postmenopáusicas / Body composition in premenopausal and postmenopausal well-trained females

Objetivo: El propósito fue analizar la influencia de las hormonas sexuales en la composición corporal de deportistas con diferente estatus hormonal. Metodología: 46mujeres eumenorreicas, 41 usuarias de píldora anticonceptiva monofásica y 16mujeres postmenopáusicas bien entrenadas participaron en el estudio. Las voluntarias realizaron un Dual-energy X-ray Absorptiometry scan (DXA) y una bioimpedancia durante la fase folicular temprana y no hormonal, verificado con una analítica. Resultados: La prueba ANCOVA no mostró diferencias ni en las variables medidas con DXA (peso, masa grasa androide y ginoide, masa grasa total y masa libre de grasa) ni en las de bioimpedancia (peso, masa grasa, masa libre de grasa y agua corporal total). Conclusión: Las hormonas sexuales parecen no influir en la composición corporal de mujeres activas. Las mujeres postmenopáusicas activas presentan una distribución de masa grasa similar a las premenopáusicas, lo que podría explicarse por el efecto positivo de la actividad física. (AU)

Purpose: The aim was to analyse the influence of sex hormones on body composition in well-trained females with different hormonal environments.Methods: Sixty-six eumenorrheic, forty-one low-dose-monophasic oral contraceptive users and sixteen postmenopausal well-trained females participated in this study. Volunteers underwent a Dual-energy X-ray Absorptiometry scan (DXA) and a bioimpedance during the early-follicular and the withdrawal phase, verified with blood samples.Results: ANCOVA test reported no differences neither in DXA measurements (weight, fat free mass, fat mass, android and gynoid fat mass) nor in bioimpedance variables (weight, fat free mass, fat mass and total body water) among study groups.Conclusion: Sex hormones seems not to influence body composition in active women. Curiously, premenopausal and postmenopausal active women present the same fat mass distribution. It could be explained by the positive effect exercise has on body composition, and this in turn on preventing cardiovascular and metabolic diseases in this population. (AU)

Development and validation of LC-MS/MS methods for the simultaneous quantification of sofosbuvir and its major metabolite (GS-331007) in blood plasma and cerebrospinal and seminal fluid: Application to a pilot clinical trial with a focus on Zika.

Zika still poses a threat to global health owing to its association with serious neurological conditions and the absence of a vaccine and treatment. Sofosbuvir, an anti-hepatitis C drug, has shown anti-Zika effects in animal and cell models. Thus, this study aimed to develop and validate novel LC-MS/MS methods for the quantification of sofosbuvir and its major metabolite (GS-331007) in human plasma and cerebrospinal (CSF) and seminal fluid (SF), and apply the methods to a pilot clinical trial. The samples were prepared by liquid-liquid extraction and separated using isocratic mode on Gemini C18 columns. Analytical detection was performed using a triple quadrupole mass spectrometer equipped with an electrospray ionization source. The validated ranges for sofosbuvir were 0.5-2,000 ng/mL (plasma) and 0.5-100 ng/mL (CSF and SF), while for the metabolite they were 2.0-2,000 ng/mL (plasma), 5.0-200 ng/mL (CSF) and 10-1,500 ng/mL (SF). The intra-day and inter-day accuracies (90.8-113.8%) and precisions (1.4-14.8%) were within the acceptance range. The developed methods fulfilled all validation parameters concerning selectivity, matrix effect, carryover, linearity, dilution integrity, precision, accuracy and stability, confirming the suitability of the method for the analysis of clinical samples.

An attempt to reduce unnecessary surgical procedures... Can ultrasound characteristics help in differentiating adenoma vs carcinoma in follicular thyroid neoplasms?

INTRODUCTION AND OBJECTIVES: Thyroid nodules frequently require ultrasound and Fine Needle Aspiration Cytology (FNAC) evaluation. However, FNA cytology does not allow differentiation between follicular adenoma and carcinoma on Bethesda type IV lesions. This situation leads to many unnecessary surgical procedures because it is not possible to assure the benignity of the lesions, even when most of the specimens correspond to adenomas or even other benign lesions. The objective is this study is to establish if there are any US characteristics that would help us to predict the risk of malignancy of nodules with a pathological diagnosis of follicular neoplasm in order to achieve a more conservative management for non-suspicious nodules. MATERIAL AND METHODS: We studied 61 nodules in 61 patients (51 women and 10 men) that underwent thyroid surgery and had histopathological results of either follicular adenoma or carcinoma. Different US characteristics of the nodules were analysed (composition, echogenicity, margin, calcification status, the presence of halo and overall observer suspicion of malignancy) and were correlated with the histopathological analysis. RESULTS: We have found a statistically significant association between the presence of calcifications, ill-defined borders and overall observer suspicion or impression (defined by well-known suspicious for malignancy ultrasonographic features, such as calcification, poorly defined margin, and a markedly hypoechoic solid nodule; and benign ultrasonographic features, such as predominantly cystic echogenic composition and the presence of a perinodular hypoechogenic halo) with follicular carcinoma. However all those features have shown low sensitivities in the present study (30%, 30% and 50%, respectively). On the other hand, the absence of halo sign has shown a sensitivity of 100% and a negative predictive value (NPV) of 100% in our study. CONCLUSIONS: The presence of calcifications, ill-defined borders and the overall impression or suspicion of malignancy associate with a higher risk for follicular carcinoma in Bethesda type IV thyroid nodules but their absence do not allow to predict benignity in these nodules. Inversely, when a halo sign lesion is observed, benign follicular neoplasm should be considered.

Un intento de reducir intervenciones quirúrgicas innecesarias... ¿Pueden ayudar las características ecográficas a diferenciar el adenoma del carcinoma en las neoplasias tiroideas foliculares? / An attempt to reduce unnecessary surgical procedures... Can ultrasound characteristics help in differentiating adenoma vs carcinoma in follicular thyroid neoplasms?

Introducción y objetivosLos nódulos tiroideos requieren con frecuencia evaluación mediante ecografía y biopsia por aspiración con aguja fina (BAAF). No obstante, la BAAF no permite la diferenciación entre adenoma y carcinoma folicular en lesiones de tipo IV según la clasificación de Bethesda. Esto provoca numerosas intervenciones quirúrgicas innecesarias porque no es posible garantizar el carácter benigno de las lesiones, ni siquiera cuando la mayoría de las muestras corresponden a adenomas o incluso a otras lesiones benignas. El objetivo de este estudio es establecer si hay características ecográficas que nos ayudarían a pronosticar el riesgo de malignidad de los nódulos con un diagnóstico anatomopatológico de neoplasia folicular para conseguir un abordaje más conservador de los nódulos sin sospecha de malignidad.Material y métodosEstudiamos 61 nódulos en 61 pacientes (51 mujeres y 10 varones) que se habían sometido a intervención quirúrgica de la glándula tiroides y tenían resultados histopatológicos de adenoma o carcinoma folicular. Se analizaron diversas características ecográficas de los nódulos (composición, ecogenicidad, borde, estado de calcificación, presencia de halo y sospecha general de malignidad según el observador) y se estableció su correlación con el análisis histopatológico.ResultadosObservamos una relación estadísticamente significativa entre el carcinoma folicular y la presencia de calcificaciones, bordes mal definidos y la sospecha o impresión general del observador (definida por sospecha clara de signos ecográficos de malignidad, como calcificación, borde mal definido y un nódulo sólido marcadamente hipoecoico; y signos ecográficos de benignidad, como composición ecogénica predominantemente quística y presencia de halo hipoecogénico perinodular). Sin embargo, todas estas características han mostrado una sensibilidad baja en el estudio que nos ocupa (30%, 30% y 50%, respectivamente)...(AU)

Introduction and objectivesThyroid nodules frequently require ultrasound and Fine Needle Aspiration Cytology (FNAC) evaluation. However, FNA cytology does not allow differentiation between follicular adenoma and carcinoma on Bethesda type IV lesions. This situation leads to many unnecessary surgical procedures because it is not possible to assure the benignity of the lesions, even when most of the specimens correspond to adenomas or even other benign lesions.The objective is this study is to establish if there are any US characteristics that would help us to predict the risk of malignancy of nodules with a pathological diagnosis of follicular neoplasm in order to achieve a more conservative management for non-suspicious nodules.Material and methodsWe studied 61 nodules in 61 patients (51 women and 10 men) that underwent thyroid surgery and had histopathological results of either follicular adenoma or carcinoma.Different US characteristics of the nodules were analysed (composition, echogenicity, margin, calcification status, the presence of halo and overall observer suspicion of malignancy) and were correlated with the histopathological analysis.ResultsWe have found a statistically significant association between the presence of calcifications, ill-defined borders and overall observer suspicion or impression (defined by well-known suspicious for malignancy ultrasonographic features, such as calcification, poorly defined margin, and a markedly hypoechoic solid nodule; and benign ultrasonographic features, such as predominantly cystic echogenic composition and the presence of a perinodular hypoechogenic halo) with follicular carcinoma. However all those features have shown low sensitivities in the present study (30%, 30% and 50%, respectively). On the other hand, the absence of halo sign has shown a sensitivity of 100% and a negative predictive value (NPV) of 100% in our study... (AU)

Preclinical development of kinetin as a safe error-prone SARS-CoV-2 antiviral able to attenuate virus-induced inflammation.

Orally available antivirals against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are necessary because of the continuous circulation of new variants that challenge immunized individuals. Because severe COVID-19 is a virus-triggered immune and inflammatory dysfunction, molecules endowed with both antiviral and anti-inflammatory activity are highly desirable. We identified here that kinetin (MB-905) inhibits the in vitro replication of SARS-CoV-2 in human hepatic and pulmonary cell lines. On infected monocytes, MB-905 reduced virus replication, IL-6 and TNFα levels. MB-905 is converted into its triphosphate nucleotide to inhibit viral RNA synthesis and induce error-prone virus replication. Coinhibition of SARS-CoV-2 exonuclease, a proofreading enzyme that corrects erroneously incorporated nucleotides during viral RNA replication, potentiated the inhibitory effect of MB-905. MB-905 shows good oral absorption, its metabolites are stable, achieving long-lasting plasma and lung concentrations, and this drug is not mutagenic nor cardiotoxic in acute and chronic treatments. SARS-CoV-2-infected hACE-mice and hamsters treated with MB-905 show decreased viral replication, lung necrosis, hemorrhage and inflammation. Because kinetin is clinically investigated for a rare genetic disease at regimens beyond the predicted concentrations of antiviral/anti-inflammatory inhibition, our investigation suggests the opportunity for the rapid clinical development of a new antiviral substance for the treatment of COVID-19.

Differential haplotype expression in class I MHC genes during SARS-CoV-2 infection of human lung cell lines

Cell entry of SARS-CoV-2 causes genome-wide disruption of the transcriptional profiles of genes and biological pathways involved in the pathogenesis of COVID-19. Expression allelic imbalance is characterized by a deviation from the Mendelian expected 1:1 expression ratio and is an important source of allele-specific heterogeneity. Expression allelic imbalance can be measured by allele-specific expression analysis (ASE) across heterozygous informative expressed single nucleotide variants (eSNVs). ASE reflects many regulatory biological phenomena that can be assessed by combining genome and transcriptome information. ASE contributes to the interindividual variability associated with disease. We aim to estimate the transcriptome-wide impact of SARS-CoV-2 infection by analyzing eSNVs. We compared ASE profiles in the human lung cell lines Calu-3, A459, and H522 before and after infection with SARS-CoV-2 using RNA-Seq experiments. We identified 34 differential ASE (DASE) sites in 13 genes (HLA-A, HLA-B, HLA-C, BRD2, EHD2, GFM2, GSPT1, HAVCR1, MAT2A, NQO2, SUPT6H, TNFRSF11A, UMPS), all of which are enriched in protein binding functions and play a role in COVID-19. Most DASE sites were assigned to the MHC class I locus and were predominantly upregulated upon infection. DASE sites in the MHC class I locus also occur in iPSC-derived airway epithelium basal cells infected with SARS-CoV-2. Using an RNA-Seq haplotype reconstruction approach, we found DASE sites and adjacent eSNVs in phase (i.e., predicted on the same DNA strand), demonstrating differential haplotype expression upon infection. We found a bias towards the expression of the HLA alleles with a higher binding affinity to SARS-CoV-2 epitopes. Independent of gene expression compensation, SARS-CoV-2 infection of human lung cell lines induces transcriptional allelic switching at the MHC loci. This suggests a response mechanism to SARS-CoV-2 infection that swaps HLA alleles with poor epitope binding affinity, an expectation supported by publicly available proteome data.

Agathisflavone, a natural biflavonoid that inhibits SARS-CoV-2 replication by targeting its proteases.

Despite the fast development of vaccines, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) still circulates through variants of concern (VoC) and escape the humoral immune response. SARS-CoV-2 has provoked over 200,000 deaths/months since its emergence and only a few antiviral drugs showed clinical benefit up to this moment. Thus, chemical structures endowed with anti-SARS-CoV-2 activity are important for continuous antiviral development and natural products represent a fruitful source of substances with biological activity. In the present study, agathisflavone (AGT), a biflavonoid from Anacardium occidentale was investigated as a candidate anti-SARS-CoV-2 compound. In silico and enzymatic analysis indicated that AGT may target mainly the viral main protease (Mpro) and not the papain-like protease (PLpro) in a non-competitive way. Cell-based assays in type II pneumocytes cell lineage (Calu-3) showed that SARS-CoV-2 is more susceptible to AGT than to apigenin (APG, monomer of AGT), in a dose-dependent manner, with an EC50 of 4.23 ± 0.21 µM and CC50 of 61.3 ± 0.1 µM and with a capacity to inhibit the level of pro-inflammatory mediator tumor necrosis factor-alpha (TNF-α). These results configure AGT as an interesting chemical scaffold for the development of novel semisynthetic antivirals against SARS-CoV-2.

Implemented occupational health surveillance limits the spread of SARS-CoV-2 Omicron at the workplace.

The global spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has put an enormous pressure on human societies, at both health and economic levels. Early diagnosis of SARS-CoV-2, the causative agent of 2019 coronavirus disease (COVID-19), has proved an efficient method to rapidly isolate positive individuals and reduce transmission rates, thus alleviating its negative impact on society's well-being and economic growth. In this work, through a coordinated and centralized effort to monitor SARS-CoV-2 circulation in companies from the State of Rio de Janeiro, Brazil, we have detected and linked an early rise of infection rates in January 2022 to the introduction of the Omicron variant of concern (VoC) (BA.1). Interestingly, when the Omicron genomic isolates were compared to correlates from public datasets, it was revealed that introduction events were multiple, with possible migration routes mapping to: Mali; Oman and United States; and Italy, Latin America, and United States. In addition, we have built a haplotype network with our genomic dataset and found no strong evidence of transmission chains, between and within companies. Considering Omicron's particularly high transmissibility, and that most of our samples (>87%) arose from 3 out of 10 companies, these findings suggest that workers from such environments were exposed to SARS-CoV-2 outside their company boundaries. Thus, using a mixed strategy in which quick molecular diagnosis finds support in comprehensive genomic analysis, we have shown that a successfully implemented occupational health program should contribute to document emerging VoC and to limit the spread of SARS-CoV-2 at the workplace.

Histological biomarkers and biometric data on trahira Hoplias malabaricus (Pisces, Characiformes, Erythrinidae): a bioindicator species in the Mearim river, Brazilian Amazon.

The objective of this study was to evaluate the levels of environmental contamination in a protected area in the Brazilian Amazon. For this, two areas were chosen along the Mearim River: the reference area (A1) and the potentially contaminated area (A2), where water samples were collected, for physicochemical and microbiological analyses, as well as specimens of Hoplias malabaricus, for the evaluation of biometric data and incidence of branchial lesions. The physicochemical analyzes of the water from both areas showed contamination (low levels of dissolved oxygen, tubidity and high iron concentrations, especially in A2). The microbiological analyzes showed that all water samples showed total coliform values higher than those acceptable by CONAMA and WHO (with higher values in A2), in addition to E. coli values higher than those allowed by legislation in A2. Regarding biometric data, male and female fishes were significantly longer and heavier in A1 during the dry and rain seasons and the gonadosomatic index also showed higher values in A1 than in A2 in both seasons. H. malabaricus showed gill lesions of minimal to moderate pathological importance in A1 and A2, indicating that specimens from both areas of the Mearim River showed biological responses to contamination. The observed changes in the water quality, bimetic parameters and the histological analyzes of the specimens of H. malabaricus directly reflect on the quality and health of the fishes in the Mearim River, and point to the urgent need for prevention and remediation of contamination in these ecosystems.

Computational and Experimental Approaches Identify Beta-Blockers as Potential SARS-CoV-2 Spike Inhibitors.

Finding antivirals for SARS-CoV-2 is still a major challenge, and many computational and experimental approaches have been employed to find a solution to this problem. While the global vaccination campaigns are the primary driver of controlling the current pandemic, orally bioavailable small-molecule drugs and biologics are critical to overcome this global issue. Improved therapeutics and prophylactics are required to treat people with circulating and emerging new variants, addressing severe infection, and people with underlying or immunocompromised conditions. The SARS-CoV-2 envelope spike is a challenging target for viral entry inhibitors. Pindolol presented a good docking score in a previous virtual screening using computational docking calculations after screening a Food and Drug Administration (FDA)-approved drug library of 2400 molecules as potential candidates to block the SARS-CoV-2 spike protein interaction with the angiotensin-converting enzyme 2 (ACE-2). Here, we expanded the computational evaluation to identify five beta-blockers against SARS-CoV-2 using several techniques, such as microscale thermophoresis, NanoDSF, and in vitro assays in different cell lines. These data identified carvedilol with a K d of 364 ± 22 nM for the SARS-CoV-2 spike and in vitro activity (EC50 of 7.57 µM, CC50 of 18.07 µM) against SARS-CoV-2 in Calu-3 cells. We have shown how we can apply multiple computational and experimental approaches to find molecules that can be further optimized to improve anti-SARS-CoV-2 activity.

Efficiency of the EmERGE Pathway to provide continuity of care for Spanish people living with medically stable HIV / Eficiencia de la vía EmERGE para brindar continuidad asistencial a personas españolas que viven con VIH clínicamente estable

Objective:Calculate the efficiency of the EmERGE Pathway of Care for medically stable people living with HIV at the Hospital Clínic-IDIBAPS, Barcelona, Spain. Methods: 546 study participants were followed between 1st July 2016 and 30th October 2019 across three HIV outpatient clinics, but the virtual clinic was closed during the second year. Unit costs were calculated, linked to mean use outpatient services per patient year, one-year before and after the implementation of EmERGE. Costs were combined with primary and secondary outcomes. Results:Annual costs across HIV-outpatient services increased by 8%: €1073 (95%CI €999–€1157) to €1158 (95%CI €1084–€1238). Annual cost of ARVs was €7,557; total annual costs increased by 1% from €8430 (95%CI €8356–8514) to €8515 (95%CI €8441–8595). Annual cost for 433 participants managed in face-to-face (F2F) clinics decreased by 5% from €958 (95%CI 905–1018) to €904 (95%CI 863–945); participants transferred from virtual to F2F outpatient clinics (V2F) increased their annual cost by a factor of 2.2, from €115 (95%CI 94–139) to €251 (95%CI 219–290). No substantive changes were observed in primary and secondary outcomes. Conclusion: EmERGE Pathway is an efficient and acceptable intervention. Increases in costs were caused by internal structural changes. The cost reduction observed in F2F clinics were off-set by the transfer of participants from the virtual to the F2F clinics due to the closure of the virtual clinic during the second year of the Study. Greater efficiencies are likely to be achieved by extending the use of the Pathway to other PLHIV.(AU)

Objetivo: Calcular la eficiencia de la vía asistencial EmERGE para personas clínicamente estables que viven con VIH en el Hospital Clínic-IDIBAPS, en Barcelona, España. Métodos: Se realizó un seguimiento a 546 participantes del estudio, entre el 1 de julio de 2016 y el 30 de octubre de 2019, en tres clínicas ambulatorias de VIH, pero la clínica virtual se cerró durante el segundo año. Se calcularon los costes unitarios, vinculados al uso medio de los servicios ambulatorios por paciente al año, un año antes y después de la implementación de EmERGE. Los costes se combinaron con criterios de valoración principales y secundarios. Resultados: Los costes anuales en los servicios ambulatorios para el VIH aumentaron un 8%: 1.073 € (IC 95%: 999-1.157 €) a 1.158 € (IC 95%: 1.084-1.238 €). El coste anual de los fármacos antirretrovirales (ARV) fue de 7.557 €; los costes anuales totales aumentaron en un 1%, de 8.430 € (IC 95%: 8.356-8.514 €) a 8.515 € (IC 95%: 8.441-8.595 €).El coste anual para 433 participantes que recibieron tratamiento en clínicas presenciales (face to face, F2F) disminuyó en un 5%, de 958 € (IC 95%: 905-1.018 €) a 904 € (IC 95%: 863-945 €); los participantes transferidos de clínicas ambulatorias virtuales (V2F) a F2F aumentaron su coste anual en un factor de 2,2, de 115 € (IC 95%: 94-139 €) a 251 € (IC 95%: 219-290 €). No se observaron cambios sustanciales en los criterios de valoración principales y secundarios. Conclusión: La vía EmERGE es un tratamiento eficaz y aceptable. Los aumentos de los costes fueron el resultado de cambios estructurales internos. La reducción de costes observada en las clínicas F2F se compensó con la transferencia de participantes de las clínicas virtuales a las F2F debido al cierre de la clínica virtual durante el segundo año del estudio. Es probable que se logre una mayor eficiencia si se amplía el uso de la vía a otras personas que viven con VIH (PVVIH).(AU)

The Role of Pyrazolopyridine Derivatives on Different Steps of Herpes Simplex Virus Type-1 In Vitro Replicative Cycle.

Herpes simplex virus type-1 (HSV-1) infection causes several disorders, and acyclovir is used as a reference compound. However, resistant strains are commonly observed. Herein, we investigate the effects of N-heterocyclic compounds (pyrazolopyridine derivatives), named ARA-04, ARA-05, and AM-57, on HSV-1 in vitro replication. We show that the 50% effective concentration (EC50) values of the compounds ARA-04, ARA-05, and AM-57 were 1.00 ± 0.10, 1.00 ± 0.05, and 0.70 ± 0.10 µM, respectively. These compounds presented high 50% cytotoxic concentration (CC50) values, which resulted in a selective index (SI) of 1000, 1000, and 857.1 for ARA-04, ARA-05, and AM-57, respectively. To gain insight into which step of the HSV-1 replication cycle these molecules would impair, we performed adsorption and penetration inhibition assays and time-of-addition experiments. Our results indicated that ARA-04 and ARA-05 affected viral adsorption, while AM-57 interfered with the virus replication during its α- and γ-phases and decreased ICP27 content during initial and late events of HSV-1 replication. In addition, we also observed that AM-57 caused a strong decrease in viral gD content, which was reinforced by in silico calculations that suggested AM-57 interacts preferentially with the viral complex between a general transcription factor and virion protein (TFIIBc-VP16). In contrast, ARA-04 and ARA-05 interact preferentially in the proteins responsible for the viral adsorption process (nectin-1 and glycoprotein). Thus, our results suggest that the 1H-pyrazolo[3,4-b]pyridine derivatives inhibit the HSV-1 replicative cycle with a novel mechanism of action, and its scaffold can be used as a template for the synthesis of promising new molecules with antiviral effects, including to reinforce the presented data herein for a limited number of molecules.

Efficiency of the EmERGE Pathway to provide continuity of care for Spanish people living with medically stable HIV.

OBJECTIVE: Calculate the efficiency of the EmERGE Pathway of Care for medically stable people living with HIV at the Hospital Clínic-IDIBAPS, Barcelona, Spain. METHODS: 546 study participants were followed between 1st July 2016 and 30th October 2019 across three HIV outpatient clinics, but the virtual clinic was closed during the second year. Unit costs were calculated, linked to mean use outpatient services per patient year, one-year before and after the implementation of EmERGE. Costs were combined with primary and secondary outcomes. RESULTS: Annual costs across HIV-outpatient services increased by 8%: €1073 (95%CI €999-€1157) to €1158 (95%CI €1084-€1238). Annual cost of ARVs was €7,557; total annual costs increased by 1% from €8430 (95%CI €8356-8514) to €8515 (95%CI €8441-8595). Annual cost for 433 participants managed in face-to-face (F2F) clinics decreased by 5% from €958 (95%CI 905-1018) to €904 (95%CI 863-945); participants transferred from virtual to F2F outpatient clinics (V2F) increased their annual cost by a factor of 2.2, from €115 (95%CI 94-139) to €251 (95%CI 219-290). No substantive changes were observed in primary and secondary outcomes. CONCLUSION: EmERGE Pathway is an efficient and acceptable intervention. Increases in costs were caused by internal structural changes. The cost reduction observed in F2F clinics were off-set by the transfer of participants from the virtual to the F2F clinics due to the closure of the virtual clinic during the second year of the Study. Greater efficiencies are likely to be achieved by extending the use of the Pathway to other PLHIV.

Associations between food portion sizes, insulin resistance, VO2 max and metabolic syndrome in European adolescents: The HELENA study.

BACKGROUND AND AIMS: This study aims to examine the associations of food portion size (PS) with markers of insulin resistance (IR) and clustered of metabolic risk score in European adolescents. METHODS: A total of 495 adolescents (53.5% females) from the Healthy Lifestyle in Europe by Nutrition in Adolescence (HELENA) study were included. The association between PS from food groups and homeostasis model assessment of insulin resistance (HOMA-IR) index, VO2 max, and metabolic risk score was assessed by multilinear regression analysis adjusting for several confounders. Analysis of covariance (ANCOVA) was used to determine the mean differences of food PS from food groups by HOMA-IR cutoff categories by using maternal education as a covariable. RESULTS: Larger PS from vegetables in both gender and milk, yoghurt, and milk beverages in males were associated with higher VO2 max, while larger PS from margarines and vegetable oils were associated with lower VO2 max (p < 0.05). Males who consumed larger PS from fish and fish products; meat substitutes, nuts, and pulses; cakes, pies, and biscuits; and sugar, honey, jams, and chocolate have a higher metabolic risk score (p < 0.05). Males with lower HOMA-IR cutoff values consumed larger PS from vegetables, milk, yoghurt, and milk beverages (p < 0.05). Females with lower HOMA-IR cutoff values consumed larger PS from breakfast cereals, while those with higher HOMA-IR cutoff values consumed larger PS from butter and animal fats (p = 0.018). CONCLUSION: The results show that larger PS from dairy products, cereals, and high energy dense foods are a significant determinant of IR and VO2 max, and larger PS from food with higher content of sugar were associated with higher metabolic risk score.

Immunogenicity of SARS-CoV-2 Trimeric Spike Protein Associated to Poly(I:C) Plus Alum.

The SARS-CoV-2 pandemic has had a social and economic impact worldwide, and vaccination is an efficient strategy for diminishing those damages. New adjuvant formulations are required for the high vaccine demands, especially adjuvant formulations that induce a Th1 phenotype. Herein we assess a vaccination strategy using a combination of Alum and polyinosinic:polycytidylic acid [Poly(I:C)] adjuvants plus the SARS-CoV-2 spike protein in a prefusion trimeric conformation by an intradermal (ID) route. We found high levels of IgG anti-spike antibodies in the serum by enzyme linked immunosorbent assay (ELISA) and high neutralizing titers against SARS-CoV-2 in vitro by neutralization assay, after two or three immunizations. By evaluating the production of IgG subtypes, as expected, we found that formulations containing Poly(I:C) induced IgG2a whereas Alum did not. The combination of these two adjuvants induced high levels of both IgG1 and IgG2a. In addition, cellular immune responses of CD4+ and CD8+ T cells producing interferon-gamma were equivalent, demonstrating that the Alum + Poly(I:C) combination supported a Th1 profile. Based on the high neutralizing titers, we evaluated B cells in the germinal centers, which are specific for receptor-binding domain (RBD) and spike, and observed that more positive B cells were induced upon the Alum + Poly(I:C) combination. Moreover, these B cells produced antibodies against both RBD and non-RBD sites. We also studied the impact of this vaccination preparation [spike protein with Alum + Poly(I:C)] in the lungs of mice challenged with inactivated SARS-CoV-2 virus. We found a production of IgG, but not IgA, and a reduction in neutrophil recruitment in the bronchoalveolar lavage fluid (BALF) of mice, suggesting that our immunization scheme reduced lung inflammation. Altogether, our data suggest that Alum and Poly(I:C) together is a possible adjuvant combination for vaccines against SARS-CoV-2 by the intradermal route.