Impact of Flash Glucose Monitoring on the Fear of Hypoglycemia Phenomenon in Adults with Type 1 Diabetes.

Objective: To assess the clinical impact of flash glucose monitoring (FGM) systems on fear of hypoglycemia (FoH) and quality of life in adults with type 1 diabetes mellitus (T1DM). Methods: Prospective quasi-experimental study with a 12-month follow-up. People with T1DM (18-80 years old) and self-monitoring by blood capillary glycemia controls were included. The FH15 questionnaire, a survey validated in Spanish in a comparable study population, was used to diagnose FoH with a cutoff point of 28 points. Results: A total of 181 participants were included, with a FoH prevalence of 69% (n = 123). A mean reduction in FH15 score of -4 points (95% confidence interval [-5.5 to -3]; P < 0.001) was observed, along with an improvement in quality of life (EsDQOL-test (Diabetes Quality of Life, Spanish version), -7 points [-10; -4], P < 0.001) and satisfaction with treatment (Diabetes Treatment Satisfaction questionnaire, self-reported version [DTSQ-s] test, +4.5 points [4; 5.5], P < 0.001). At the end of the follow-up, 64.2% of the participants saw an improved FoH intensity, compared to 35.8% who scored the same or higher. This improvement in FoH status was associated with a higher time-in-range at the end of the follow-up (P = 0.003), as well as a lower time spent in hyperglycemia (P = 0.005). In addition, it was linked to participants with a high baseline FoH levels (P < 0.001) and those who were university degree holders (P = 0.07). Conclusions: FGM is associated with an overall reduction of FoH in adults with T1DM and with an increase in their quality of life. Nevertheless, a significant percentage of patients may experience an increase of this phenomenon leading to clinical repercussions and a profound impact on quality of life.

Aplicabilidad del índice de fragilidad hepática en pacientes cirróticos evaluados para trasplante / Applicability of the liver frailty index in cirrhotic patients evaluated for transplantation

Introducción. En las últimas décadas se han desarrollado diferentes scores y modelos para predecir el pronóstico en pacientes con enfermedad hepática crónica avanzada. Los más reconocidos y utilizados son el sistema de estadificación de Child-Pugh (CP) y el score de MELD, pero estos carecen de herramientas para evaluar objetivamente otros factores pronósticos. Por este motivo, se ha incorporado el concepto de fragilidad a la hepatología clínica. El objetivo de este artículo es examinar la aplicabilidad del índice de fragilidad hepática (IFH) en pacientes con cirrosis evaluados para trasplante hepático en Uruguay. Metodología. Estudio observacional, descriptivo y retrospectivo en el Servicio de Enfermedades Hepáticas del Hospital Central de las Fuerzas Armadas (HCFFAA) de enero de 2018 a diciembre de 2021. Resultados. Se evaluaron un total de 78 pacientes, excluyéndose 19 de estos, culminando con una muestra final de 59 pacientes. La edad media fue de 52 años, siendo el 66 % hombres. La principal etiología de la cirrosis fue la alcohólica, y la comorbilidad más frecuente fue el sobrepeso/obesidad (66 %). La media de IFH fue de 4,03 ± 0,45. El 90 % de los pacientes eran prefrágiles, el 10 % frágiles y ningún paciente fue clasificado como no frágil. El 76 % presentaba un estadio avanzado de la enfermedad al momento de la evaluación 42 % CP estadio B, 34 % CP C, 24 % CP A, con una media de MELD-Na de 17,8 ± 7,6. El 17 % tuvo complicaciones infecciosas. La mortalidad global (n=78) fue del 12 %, y la de los pacientes con IFH calculado fue del 22 %. Conclusiones. El cálculo del IFH es realizable en cirróticos como herramienta objetiva que brinda una mirada integral del paciente. A mayor severidad de la cirrosis, mayor es el IFH. Sin embargo, este índice no parece ser un predictor de la eventual realización del trasplante hepático, ni de muerte en lista de espera en nuestros pacientes.

Introduction. In recent decades, several scores and models have been proposed to predict prognosis in patients with advanced chronic liver disease. The most recognized and used are the Child-Pugh (CP) and the Model for End-stage Liver Disease (MELD) scores, but they lack tools to objectively evaluate other prognostic factors. For this reason, the concept of fragility has been incorporated into clinical hepatology. The objective of this study was to evaluate the applicability of the liver frailty index (LFI) in patients with cirrhosis evaluated for liver transplantation in Uruguay. Methodology. Observational, descriptive and retrospective study at the Hospital Central de las Fuerzas Armadas (HCFFAA) Liver Disease Service from January 2018 to December 2021. Results. A total of 78 patients were evaluated, 19 were excluded, culminating in a final sample of 59 patients. The mean age was 52 years, with 66% being men. The main etiology of cirrhosis was alcoholic and the most frequent comorbidity was overweight/obesity (66%). The mean LFI was 4.03 ± 0.45. 90% of patients were pre-fragile, 10% were fragile, and no patient was classified as non-fragile. 76% had an advanced stage of the disease at the time of evaluation: 42% CP stage B, 34% CP C, 24% CP A, with a mean MELD-Na of 17.8 ± 7.6. 17% had infectious complications. Overall mortality (n=78) was 12%, and that of patients with calculated LFI was 22%. Conclusions. The LFI can be calculated in cirrhotic patients, and it is an objective tool that provides a comprehensive view of the patient. LFI depends on the severity of the cirrhosis. However, this index is not a predictor of liver transplantation or death on the waiting list in our patients.

Impact of an Evidence-Based Prioritization System and Electronic Consultation in Early Diagnosis of Colorectal Cancer.

(1) Background: Colorectal cancer (CRC) is one of the most common causes of cancer. Timely diagnosis is critical, with even minor delays impacting prognosis. Primary care providers face obstacles in accessing specialist care. This study investigates the impact of implementing an electronic consultation (eConsult) system combined with a specific prioritization system on CRC diagnosis delay and tumor staging. (2) Methods: The study analyzes 245 CRC patients from November 2019 to February 2022, comparing those referred before and after the eConsult system's implementation during the COVID-19 pandemic. Data on referral reasons, pathways, diagnosis delays, and staging were collected. Multivariate analysis aimed to identify independent risk factors for advanced staging at diagnosis. (3) Results: The eConsult system significantly reduced CRC diagnosis delay from 68 to 26 days. The majority of patients referred via eConsult presented with symptoms. Despite expedited diagnoses, no discernible difference in CRC staging emerged between eConsult and traditional referrals. Notably, patients from screening programs or with a positive fecal immunochemical test (FIT) experienced earlier-stage diagnoses. A positive FIT without symptoms and being a never-smoker emerged as protective factors against advanced-stage CRC. (4) Conclusions: This study highlights eConsult's role in reducing CRC diagnosis delay, improving diagnostic efficiency and prioritizing urgent cases, emphasizing FIT effectiveness.

Enzyme-less nanopore detection of post-translational modifications within long polypeptides.

Means to analyse cellular proteins and their millions of variants at the single-molecule level would uncover substantial information previously unknown to biology. Nanopore technology, which underpins long-read DNA and RNA sequencing, holds potential for full-length proteoform identification. We use electro-osmosis in an engineered charge-selective nanopore for the non-enzymatic capture, unfolding and translocation of individual polypeptides of more than 1,200 residues. Unlabelled thioredoxin polyproteins undergo transport through the nanopore, with directional co-translocational unfolding occurring unit by unit from either the C or N terminus. Chaotropic reagents at non-denaturing concentrations accelerate the analysis. By monitoring the ionic current flowing through the nanopore, we locate post-translational modifications deep within the polypeptide chains, laying the groundwork for compiling inventories of the proteoforms in cells and tissues.

Enfoques teóricos y metodológicos sobre las emociones en un contexto de cooperación internacional / Theoretical and methodological approaches to emotions in a context of International Cooperation

El papel de las emociones es crucial en la vida cotidiana y en las experiencias de los cooperantes sociosanitarios, específicamente en el contexto de la cooperación internacional. Aunque las ciencias sociales y sociosanitarias han empezado a explorar las emociones en este ámbito, su estudio es aun relativamente joven. Se realiza una revisión teórica desde un enfoque interdisciplinario, abarcando tanto las Ciencias de la Salud como las Ciencias Sociales. Este enfoque mixto proporciona una visión más completa de la compleja relación entre las emociones y la cooperación internacional. Además de presentar un marco teórico, se revisan diferentes enfoques metodológicos, desde técnicas tradicionales que analizan discursos y expresiones emocionales hasta propuestas innovadoras como el Modelo FEI (Felt, Expressed and Identified emotions), que aborda la complejidad emocional. Este trabajo contribuye al desarrollo del campo, ofreciendo una base teórica y analítica para futuros estudios en esta área multidimensional.(AU)

The role of emotions is crucial in everyday life and the experiences of healthcare cooperators, specifically in the context of international cooperation. Although the social and healthcare sciences have begun to explore emotions in this field, their study is still relatively young. A theoretical review is conducted from an interdisciplinary perspective, encompassing both Health Sciences and Social Sciences. This mixed approach provides a more comprehensive understanding of the complex relationship between emotions and international cooperation. In addition to presenting a theoretical framework, various methodological approaches are reviewed, from traditional techniques analysing emotional discourses and expressions to innovative proposals such as the Felt, Expressed, and Identified emotions (FEI) Model, which addresses emotional complexity. This work contributes to the development of the field, offering a theoretical and analytical foundation for future studies in this multidimensional area.(AU)

Development and Validation of SERR Scale for Detecting Extremism and Religious Radicalism.

BACKGROUND: In line with the growing religious, political and social violence around the world, this study evaluates the psychometric properties of a new scale for Detecting Extremism and Religious Radicalism (SERR), a self-report measure of extremism and religious radicalism. METHOD: Factor analyses, validity and reliability data were collected using a sample of 1985 participants from 58 different cities around Spain who self-identified as actively practicing their religion. RESULTS: Statistical analyses of SERR scores yielded acceptable internal consistency scores (ω > .74) and confirmed key factors associated with radicalism and extremism. The structure of the scale was confirmed as two-dimensional (χ2(64) = 361.22, p < .001, CFI = 0.976, TLI = 0.970, RMSEA (90% CI) = 0.089 (0.080 - 0.098), SRMR = 0.064). Scores for the extremism dimension correlated significantly and positively with scores for the same factor on the Activism and Radicalism Intention Scales (r = .32, p < .001, n = 139). CONCLUSIONS: The SERR demonstrates adequate reliability and validity for evaluating the degree of extremism and radicalism in Christian/Catholic and Muslim believers.

Development and Validation of SERR Scale for Detecting Extremism and Religious Radicalism / Desarrollo y Validación de la Escala SERR para la Detección del Extremismo y Radicalismo Religioso

Background: In liiine with the growing religious, political and social violence around the world, this study evaluates the psychometric properties of a new scale for Detecting Extremism and Religious Radicalism (SERR), a self-report measure of extremism and religious radicalism. Method: Factor analyses, validity and reliability data were collected using a sample of 1985 participants from 58 different cities around Spain who self-identified as actively practicing their religion. Results: Statistical analyses of SERR scores yielded acceptable internal consistency scores (ω > .74) and confirmed key factors associated with radicalism and extremism. The structure of the scale was confirmed as two dimensional (χ²(64) = 361.22, p < .001, CFI = 0.976, TLI = 0.970, RMSEA (90% CI) = 0.089 (0.080 - 0.098), SRMR = 0.064). Scores for the extremism dimension correlated significantly and positively with scores for the same factor on the Activism and Radicalism Intention Scales (r = .32, p < .001, n = 139). Conclusions: The SERR demonstrates adequate reliability and validity for evaluating the degree of extremism and radicalism in Christian/Catholic and Muslim believers.(AU)

Antecedentes: en consonancia con la creciente violencia religiosa, política y social en todo el mundo, este estudio evalúa las propiedades psicométricas de la escala para la detección del extremismo y el radicalismo religioso (SERR), una medida de autoinforme del extremismo y el radicalismo religioso. Método: los análisis factoriales y las evidencias de validez y fiabilidad se recopilaron utilizando una muestra de 1985 participantes de 58 ciudades distintas de España que se identificaron a sí mismos como practicantes activos de su religión. Resultados: los análisis estadísticos de las puntuaciones SERR arrojaron puntuaciones de consistencia interna aceptables (ω > .74) y confirmaron los factores clave asociados con el radicalismo y el extremismo. La estructura de la escala se confirmó como bidimensional (χ²(64) = 361.22, p < .001, CFI = 0.976, TLI = 0.970, RMSEA (90% CI) = 0.089 (0.080 - 0.098), SRMR = 0.064). Las puntuaciones de la dimensión de extremismo se correlacionaron significativa y positivamente con las puntuaciones del mismo factor en la Escala de Intención de Activismo y Radicalismo (r = 0,32, p < 0.001, n = 139). Conclusiones: los autores concluyen que el SERR muestra una fiabilidad y validez adecuadas para evaluar el grado de extremismo y radicalismo en creyentes cristianos/católicos y musulmanes.(AU)

Spanish National Registry of Paediatric Coeliac Disease: Changes in the Clinical Presentation in the 21st Century.

OBJECTIVES: Over the last several decades, there has been a tendency towards a predominance of less symptomatic forms of coeliac disease (CD) and an increase in the patient age at diagnosis. This study aimed to assess the clinical presentation and diagnostic process of paediatric CD in Spain. METHODS: A nationwide prospective, observational, multicentre registry of new paediatric CD cases was conducted from January 2011 to June 2017. The data regarding demographic variables, type of birth, breast-feeding history, family history of CD, symptoms, height and weight, associated conditions, serological markers, human leukocyte antigen (HLA) phenotype, and histopathological findings were collected. RESULTS: In total, 4838 cases (61% girls) from 73 centres were registered. The median age at diagnosis was 4 years. Gastrointestinal symptoms were detected in 71.4% of the patients, and diarrhoea was the most frequent symptom (45.9%). The most common clinical presentation was the classical form (65.1%) whereas 9.8% ofthe patients were asymptomatic. There was a trend towards an increase in the age at diagnosis, proportion of asymptomatic CD cases, and usage of anti-deamidated gliadin peptide antibodies and HLA typing for CD diagnosis. There was, however, a decreasing trend in the proportion of patients undergoing biopsies. Some of these significant trend changes may reflect the effects of the 2012 ESPGHAN diagnosis guidelines. CONCLUSIONS: Paediatric CD in Spain is evolving in the same direction as in the rest of Europe, although classical CD remains the most common presentation form, and the age at diagnosis remains relatively low.

Update in Diagnostics of Toscana Virus Infection in a Hyperendemic Region (Southern Spain).

The sandfly fever Toscana virus (TOSV, genus Phlebovirus, family Phenuiviridae) is endemic in Mediterranean countries. In Spain, phylogenetic studies of TOSV strains demonstrated that a genotype, different from the Italian, was circulating. This update reports 107 cases of TOSV neurological infection detected in Andalusia from 1988 to 2020, by viral culture, serology and/or RT-PCR. Most cases were located in Granada province, a hyperendemic region. TOSV neurological infection may be underdiagnosed since few laboratories include this virus in their portfolio. This work presents a reliable automated method, validated for the detection of the main viruses involved in acute meningitis and encephalitis, including the arboviruses TOSV and West Nile virus. This assay solves the need for multiple molecular platforms for different viruses and thus, improves the time to results for these syndromes, which require a rapid and efficient diagnostic approach.

Influence of weight status on bone mineral content measured by DXA in children.

INTRODUCTION: Childhood obesity is a public health problem with repercussions in later life. As tissue formation peaks in childhood we determined how weight status influences bone mineral content. MATERIAL AND METHODS: We studied 553 children aged 4-18 years over 10 years (46.8% girls). We measured age, weight, height and through bone densitometry (DXA), bone mineral content (BMC), bone mineral density (BMD), and waist, arm and hip circumferences. The patients were divided into groups using the body mass index z-score: underweight, normal weight, overweight, obese and very obese. RESULTS: BMC and BMD values were highest in the normal-weight and overweight groups. Logistic regression showed bone mineralization was inversely associated with waist circumference, the association being positive for weight and age. No differences were found according to sex. DISCUSSION: Studies of the relationship between weight and bone mineralization report contradictory results, often because of different study designs. Moreover, studies in children are either few or with small samples. Our findings in a large sample show the importance of weight status in bone mineralization given the risk of bone fractures or osteoporosis. CONCLUSIONS: Weight status influenced bone mineralization. BMC and BMD decreased in children with a higher degree of obesity. Waist circumference correlated negatively with bone mineralization.

Evaluation of a Trio Toscana Virus Real-Time RT-PCR Assay Targeting Three Genomic Regions within Nucleoprotein Gene.

Toscana virus (TOSV) can cause central nervous system infections in both residents of and travelers to Mediterranean countries. Data mining identified three real-time RT-qPCR assays for detecting TOSV RNA targeting non-overlapping regions in the nucleoprotein gene. Here, they were combined to create a multi-region assay named Trio TOSV RT-qPCR consisting of six primers and three probes. In this study, (i) we evaluated in silico the three RT-qPCR assays available in the literature for TOSV detection, (ii) we combined the three systems to create the Trio TOSV RT-qPCR, (iii) we assessed the specificity and sensitivity of the three monoplex assays versus the Trio TOSV RT-qPCR assay, and (iv) we compared the performance of the Trio TOSV RT-qPCR assay with one of the reference monoplex assays on clinical samples. In conclusion, the Trio TOSV RT-qPCR assay performs equally or better than the three monoplex assays; therefore, it provides a robust assay that can be used for both research and diagnostic purposes.

Spanish Pediatric Inflammatory Bowel Disease Diagnostic Delay Registry: SPIDER Study From Sociedad Española de Gastroenterología, Hepatología y Nutrición Pediátrica.

Background and Aims: Diagnostic delay (DD) is especially relevant in children with inflammatory bowel disease, leading to potential complications. We examined the intervals and factors for DD in the pediatric population of Spain. Methods: We conducted a multicentric prospective study, including 149 pediatric inflammatory bowel disease patients, obtaining clinical, anthropometric, and biochemical data. Time to diagnosis (TD) was divided into several intervals to identify those where the DD was longer and find the variables that prolonged those intervals. Missed opportunities for diagnosis (MODs) were also identified. Results: Overall TD was 4.4 months (interquartile range [IQR] 2.6-10.4), being significantly higher in Crohn's disease (CD) than in ulcerative colitis (UC) (6.3 [IQR 3.3-12.3] vs. 3 [IQR 1.6-5.6] months, p = 0.0001). Time from the visit to the first physician until referral to a pediatric gastroenterologist was the main contributor to TD (2.4 months [IQR 1.03-7.17] in CD vs. 0.83 months [IQR 0.30-2.50] in UC, p = 0.0001). One hundred and ten patients (78.3%) visited more than one physician (29.9% to 4 or more), and 16.3% visited the same physician more than six times before being assessed by the pediatric gastroenterologist. The number of MODs was significantly higher in CD than that in UC patients: 4 MODs (IQR 2-7) vs. 2 MODs ([IQR 1-5], p = 0.003). Referral by pediatricians from hospital care allowed earlier IBD diagnosis (odds ratio 3.2 [95% confidence interval 1.1-8.9], p = 0.025). Conclusions: TD and DD were significantly higher in CD than those in UC. IBD patients (especially those with CD) undergo a large number of medical visits until the final diagnosis.

COVID-19 Prevalence, Risk Perceptions, and Preventive Behavior in Asymptomatic Latino Population: A Cross-Sectional Study.

Aims To determine the prevalence, level of coronavirus disease 2019 (COVID-19) risk perception attitude and preventive behavior implemented by the Latino population in the United States of America (USA). Methods This cross-sectional study was conducted between July 25 and August 25, 2020, and included asymptomatic Latino participants (n=410) with no current/previous COVID-19 within a religious community in Maryland. Participants answered a questionnaire consisting of three components: patient demographics/socioeconomic status, COVID-19 risk perception, and precautionary behavior. Additionally, a focused history taking and physical examination were performed, and nasal swabs for COVID-19 testing were obtained. Results Around 80% of our study population was 35 years and older, considerably healthy, with only about a third reporting history of chronic disease (~80%); most were females (~66%). Of our participants, 90% lived under poverty; only ~6% had made it to college. When asked about the likelihood of acquiring COVID-19, 97.3% stated they have a low risk of getting infected. However, as we asked about the risk of individuals living in their neighborhood, state, and country, the rates changed to moderate to high (78.4%, 86.3%, and 86.6%, respectively). When asked about preventive behavior, 71.2% stated they never wear masks and 85.4% mentioned they never keep social distance. Additionally, 76 (18.5%) tested positive for COVID-19, whereas 64 (84.2%) developed symptoms at follow-up, 57 (75%) were hospitalized, and 4 (5.2%) died. Conclusions Our study identified inadequate COVID-19 threat perception and lack of engagement in preventive behavior among a group of Latinos living in the USA. We believe that Latino communities across the USA are at markedly high risk of acquiring, spreading, and dying of COVID-19.

Therapeutic Options for COVID-19: A Review.

An acute respiratory disease caused by a novel coronavirus [severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), previously known as 2019-nCoV], the coronavirus disease 2019 (COVID-19) was first detected in Wuhan, China. Since then, the virus has spread rapidly worldwide leading to a global public health crisis. Due to its devastating effect on public health, it is crucial to identify a viable therapeutic option to mitigate the damage the disease causes. In spite of various governments implementing aggressive global lock-down and quarantine protocols, the number of cases continues to follow an upward trend. At present, the therapeutic strategies are supportive or preventative, focusing on reducing transmission. Given the gravity of the situation, we aim to explore the drugs that have been tried so far and their efficacy when applied in clinical trials. Since newer interventions would take months to years to develop, by looking at the pool of existing therapeutic options, including remdesivir (RDV), plasma exchange or cytapheresis, hydroxychloroquine, baricitinib, and lopinavir (LPV), we have tried to detail the principles behind their use to treat COVID-19, current application, and adverse effects. Many coronaviruses have a highly mutable single-stranded RNA genome and hence discovering new drugs against the virus is going to be challenging owing to the possible viral genetic recombination. Extensive research is still needed to safely advocate the efficacy of the currently available therapeutic options.

What Should Be Known by a Urologist About the Medical Management of COVID-19's Patients?

PURPOSE OF REVIEW: The alarming number of confirmed COVID-19 cases put a strain on the healthcare systems, which had to reallocate human and technical resources to respond to the emergency. Many urologists became integrated into multidisciplinary teams, dealing with this respiratory illness and its unknown management. It aims to summarize the epidemiological, clinical, diagnostical, and therapeutical characteristics of COVID-19, from a practical perspective, to ease COVID-19 management to non-physician staff. RECENT FINDINGS: We performed a narrative review of the literature regarding COVID-19, updated to May 8th, 2020, at PubMed and COVID resource platforms of the main scientific editorials. COVID-19, characterized by fever, myalgias, dyspnea, and dry cough, varies widely from asymptomatic infection to death. Arrhythmias and thrombotic events are prevalent. Lymphopenia and inflammatory reactant elevation on laboratory, as well as bilateral and peripheral ground-glass opacities or consolidations on X-Ray, are usually found in its assessment. Little is known about SARS-CoV-2 immunology. To date, no therapy has demonstrated efficacy in COVID-19. Of-level or compassionate-use therapies are prescribed in the context of clinical trials. We should become familiar with specific adverse events and pharmacological interactions. The COVID-19 pandemic has paralyzed the urological activity, and its long-term consequences are unpredictable. Despite not being used to deal with respiratory diseases, the urologists become easily qualified to manage COVID-19 by following protocols and being integrated into multidisciplinary teams, helping to overcome the pandemic.

Sample pooling for SARS-CoV-2 RT-PCR screening.

OBJECTIVE: To evaluate the efficacy of sample pooling compared to the individual analysis for the diagnosis of coronavirus disease 2019 (COVID-19) by using different commercial platforms for nucleic acid extraction and amplification. METHODS: A total of 3519 nasopharyngeal samples received at nine Spanish clinical microbiology laboratories were processed individually and in pools (342 pools of ten samples and 11 pools of nine samples) according to the existing methodology in place at each centre. RESULTS: We found that 253 pools (2519 samples) were negative and 99 pools (990 samples) were positive; with 241 positive samples (6.85%), our pooling strategy would have saved 2167 PCR tests. For 29 pools (made out of 290 samples), we found discordant results when compared to their correspondent individual samples, as follows: in 22 of 29 pools (28 samples), minor discordances were found; for seven pools (7 samples), we found major discordances. Sensitivity, specificity and positive and negative predictive values for pooling were 97.10% (95% confidence interval (CI), 94.11-98.82), 100%, 100% and 99.79% (95% CI, 99.56-99.90) respectively; accuracy was 99.80% (95% CI, 99.59-99.92), and the kappa concordant coefficient was 0.984. The dilution of samples in our pooling strategy resulted in a median loss of 2.87 (95% CI, 2.46-3.28) cycle threshold (Ct) for E gene, 3.36 (95% CI, 2.89-3.85) Ct for the RdRP gene and 2.99 (95% CI, 2.56-3.43) Ct for the N gene. CONCLUSIONS: We found a high efficiency of pooling strategies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA testing across different RNA extraction and amplification platforms, with excellent performance in terms of sensitivity, specificity and positive and negative predictive values.

Protein Splicing Activity of the Haloferax volcanii PolB-c Intein Is Sensitive to Homing Endonuclease Domain Mutations.

Inteins are selfish genetic elements residing in open reading frames that can splice post-translationally, resulting in the ligation of an uninterrupted, functional protein. Like other inteins, the DNA polymerase B (PolB) intein of the halophilic archaeon Haloferax volcanii has an active homing endonuclease (HEN) domain, facilitating its horizontal transmission. Previous work has shown that the presence of the PolB intein exerts a significant fitness cost on the organism compared to an intein-free isogenic H. volcanii. Here, we show that mutation of a conserved residue in the HEN domain not only reduces intein homing but also slows growth. Surprisingly, although this mutation is far from the protein splicing active site, it also significantly reduces in vitro protein splicing. Moreover, two additional HEN domain mutations, which could not be introduced to H. volcanii, presumably due to lethality, also eliminate protein splicing activity in vitro. These results suggest an interplay between HEN residues and the protein splicing domain, despite an over 35 Å separation in a PolB intein homology model. The combination of in vivo and in vitro evidence strongly supports a model of codependence between the self-splicing domain and the HEN domain that has been alluded to by previous in vitro studies of protein splicing with HEN domain-containing inteins.