Probiotic improves symptomatic and viral clearance in Covid19 outpatients: a randomized, quadruple-blinded, placebo-controlled trial.

Intestinal bacteria may influence lung homeostasis via the gut-lung axis. We conducted a single-center, quadruple-blinded, randomized trial in adult symptomatic Coronavirus Disease 2019 (Covid19) outpatients. Subjects were allocated 1:1 to probiotic formula (strains Lactiplantibacillus plantarum KABP022, KABP023, and KAPB033, plus strain Pediococcus acidilactici KABP021, totaling 2 × 109 colony-forming units (CFU)) or placebo, for 30 days. Co-primary endpoints included: i) proportion of patients in complete symptomatic and viral remission; ii) proportion progressing to moderate or severe disease with hospitalization, or death; and iii) days on Intensive Care Unit (ICU). Three hundred subjects were randomized (median age 37.0 years [range 18 to 60], 161 [53.7%] women, 126 [42.0%] having known metabolic risk factors), and 293 completed the study (97.7%). Complete remission was achieved by 78 of 147 (53.1%) in probiotic group compared to 41 of 146 (28.1%) in placebo (RR: 1.89 [95 CI 1.40-2.55]; P < .001), significant after multiplicity correction. No hospitalizations or deaths occurred during the study, precluding the assessment of remaining co-primary outcomes. Probiotic supplementation was well-tolerated and reduced nasopharyngeal viral load, lung infiltrates and duration of both digestive and non-digestive symptoms, compared to placebo. No significant compositional changes were detected in fecal microbiota between probiotic and placebo, but probiotic supplementation significantly increased specific IgM and IgG against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV2) compared to placebo. It is thus hypothesized this probiotic primarily acts by interacting with the host's immune system rather than changing colonic microbiota composition. Future studies should replicate these findings and elucidate its mechanism of action (Registration: NCT04517422).Abbreviations: AE: Adverse Event; BMI: Body Mass Index; CONSORT: CONsolidated Standards of Reporting Trials; CFU: Colony-Forming Units; eDRF: Electronic Daily Report Form; GLA: Gut-Lung Axis; GSRS: Gastrointestinal Symptoms Rating Scale; hsCRP: High-sensitivity C-Reactive Protein; HR: Hazard Ratio; ICU: Intensive Care Unit; OR: Odds Ratio; PCoA: Principal Coordinate Analysis; RR: Relative Risk; RT-qPCR: Real-Time Quantitative Polymerase Chain Reaction; SARS-CoV2: Severe acute respiratory syndrome coronavirus 2; SpO2: Peripheral Oxygen Saturation; WHO: World Health Organization.

Probiotic Effect on SARS‐CoV2 Immunity Is Associated to Type‐1 Interferons: A Post‐Hoc Analysis of a Randomized, Placebo‐Controlled Trial

Objectives and Hypothesis Gut microbiota has been reported to protect from lung viral infection in animal models by stimulating type‐I interferon signaling. Type‐I interferons can have direct antiviral activity while also stimulating antibody‐producing B cells. Antibodies against Severe Acute Respiratory Syndrome coronavirus 2 (SARS‐CoV2) have been correlated to faster infection clearance and protection against reinfection. A specific 4‐strain probiotic combination (Pediococcus acidilactici CECT7483 plus Lactoplantibacillus plantarum CECT7484, CECT7485 and CECT30292) was recently studied in a randomized, quadruple‐blinded, placebo‐controlled trial in 300 SARS‐CoV2‐infected, symptomatic ambulatory patients (NCT04517422). Study subjects did not receive corticosteroids or antivirals. Compared to placebo, probiotic intervention (2x109cfu/day for 30 days) achieved faster symptom clearance and increased SARS‐CoV2‐specific immunoglobulins M and G (IgM and IgG). We hypothesize these effects could be related to increased type‐I interferon signaling. Methods A random subset of 70 subjects (35 probiotic and 35 placebo) was selected out of the 300 participants in the clinical study. Stored serum samples collected on day 0 (baseline), 15 and 30 (end of intervention) were analyzed for interferon‐alpha 2a (IFNa) and interferon‐beta 1a (IFNb) using enzyme‐linked immunosorbent assay. Demographic data, baseline viral load, symptom duration and SARS‐CoV2‐specific IgM and IgG serum titers were retrieved from patient case report forms. Differences between probiotic and placebo were compared using Mann‐Whitney or Chi‐squared tests, as appropriate. Correlations between type‐I interferons and other variables were assessed by Spearman correlation. All procedures on study subjects had been approved by the Internal Review Board of Hospital General Dr. Manuel Gea (Mexico City) and adhered to Helsinki Declaration. Summary of Results In the selected subset of patients, study groups were comparable at baseline (all p>0.10, Table 1). Probiotic treatment was associated to a larger increase of IFNa on day 30 (p=0.007) and of IFNb on days 15 and 30 (both p<0.001), compared to placebo (Figure 1). Similarly, SARS‐CoV2‐specific IgM and IgG were higher on days 15 and 30 (all p<0.001) and duration of fever, cough, headache and myalgia were shorter (all p<0.05) in probiotic compared to placebo. Increase in IFNb across the study correlated to increase in SARS‐CoV2‐specific IgM (rho=0.55, p<0.001) and IgG (rho=0.61, p<0.001), and inversely correlated to duration of cough (rho=‐0.26, p=0.033) and fever (rho=‐0.24, p=0.042). Conversely, increase in IFNa correlated to increase in SARS‐CoV2‐specific IgM (rho=0.33, p=0.005) only. Neither increase in IFNa nor IFNb correlated to age, number of metabolic comorbidities or days from onset of symptoms. Conclusion Compared to placebo, oral administration of a specific 4‐strain probiotic combination resulted in significant increase in type‐I interferons in serum, especially IFNb, correlating to higher SARS‐CoV2‐specific IgM and IgG antibodies and faster clearance of some symptoms.

Inflammatory pathways in COVID-19: Mechanism and therapeutic interventions.

The 2019 coronavirus disease (COVID-19) pandemic has become a global crisis. In the immunopathogenesis of COVID-19, SARS-CoV-2 infection induces an excessive inflammatory response in patients, causing an inflammatory cytokine storm in severe cases. Cytokine storm leads to acute respiratory distress syndrome, pulmonary and other multiorgan failure, which is an important cause of COVID-19 progression and even death. Among them, activation of inflammatory pathways is a major factor in generating cytokine storms and causing dysregulated immune responses, which is closely related to the severity of viral infection. Therefore, elucidation of the inflammatory signaling pathway of SARS-CoV-2 is important in providing otential therapeutic targets and treatment strategies against COVID-19. Here, we discuss the major inflammatory pathways in the pathogenesis of COVID-19, including induction, function, and downstream signaling, as well as existing and potential interventions targeting these cytokines or related signaling pathways. We believe that a comprehensive understanding of the regulatory pathways of COVID-19 immune dysregulation and inflammation will help develop better clinical therapy strategies to effectively control inflammatory diseases, such as COVID-19.

Cell deaths: Involvement in the pathogenesis and intervention therapy of COVID-19.

The current pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has dramatically influenced various aspects of the world. It is urgent to thoroughly study pathology and underlying mechanisms for developing effective strategies to prevent and treat this threatening disease. It is universally acknowledged that cell death and cell autophagy are essential and crucial to maintaining host homeostasis and participating in disease pathogenesis. At present, more than twenty different types of cell death have been discovered, some parts of which have been fully understood, whereas some of which need more investigation. Increasing studies have indicated that cell death and cell autophagy caused by coronavirus might play an important role in virus infection and pathogenicity. However, the knowledge of the interactions and related mechanisms of SARS-CoV-2 between cell death and cell autophagy lacks systematic elucidation. Therefore, in this review, we comprehensively delineate how SARS-CoV-2 manipulates diverse cell death (including apoptosis, necroptosis, pyroptosis, ferroptosis, and NETosis) and cell autophagy for itself benefits, which is simultaneously involved in the occurrence and progression of COVID-19, aiming to provide a reasonable basis for the existing interventions and further development of novel therapies.

Factors associated with death in children with COVID-19 in Mexico.

INTRODUCTION: Most children affected by SARS-CoV-2 are reported to be asymptomatic, and COVID-19-related mortality in them is low; in Mexico, there is a lack of information on the subject in this population group. OBJECTIVE: To assess the risk factors associated with mortality in Mexican children with COVID-19. METHOD: Secondary analysis of the General Directorate of Epidemiology database. Children younger than 19 years, in whom SARS-CoV-2 infection was confirmed by RT-PCR, were included. RESULTS: 1443 children were included. Median age was eight years; 3.3 % were admitted to the intensive care unit, 1.8 % required assisted mechanical ventilation, and mortality was 1.9 %. In multivariate models, the development of pneumonia was the main risk factor for mortality, with an odds ratio (OR) of 6.45 (95 % CI: 1.99, 20.89); patients who required intubation had an OR of 8.75 (95 % CI: 3.23, 23.7). CONCLUSIONS: Children with COVID-19 exhibit high mortality in Mexico, and avoiding pneumonia should therefore be tried in them, especially in children younger than four years, with cardiovascular risk or immunosuppression. INTRODUCCIÓN: Se informa que la mayoría de los niños afectados por SARS-CoV-2 cursan asintomáticos y que en ellos la mortalidad por COVID-19 es baja; en México se desconoce la información al respecto en este grupo de la población. ­. OBJETIVO: Evaluar los factores de riesgo asociados a mortalidad en niños mexicanos con COVID-19. MÉTODO: Análisis secundario de la base de datos de la Dirección General de Epidemiología. Se incluyeron niños menores de 19 años, en quienes se confirmó SARS-CoV-2 mediante RT-PCR. RESULTADOS: Se incluyeron 1443 niños. La mediana de edad fue de ocho años; 3.3 % ingresó a la unidad de cuidados intensivos, 1.8 % requirió ventilación mecánica asistida y la mortalidad fue de 1.9 %. En los modelos multivariados, el desarrollo de neumonía constituyó el principal factor de riesgo de mortalidad, con razón de momios (RM) de 6.45 (IC 95 % 1.99, 20.89); los pacientes que requirieron intubación tuvieron RM de 8.75 (IC 95 % 3.23, 23.7). CONCLUSIONES: Los niños con COVID 19 tienen alta mortalidad en México, por lo que en ellos se debe procurar evitar la neumonía, especialmente en los menores de cuatro años, con riesgo cardiovascular o inmunosupresión.

Mortality risk factors in Mexican children with COVID-19 / Factores asociados a muerte en niños con COVID-19 en México

Resumen Introducción: Se informa que la mayoría de los niños afectados por SARS-CoV-2 cursan asintomáticos y que en ellos la mortalidad por COVID-19 es baja; en México se desconoce la información al respecto en este grupo de la población. Objetivo: Evaluar los factores de riesgo asociados a mortalidad en niños mexicanos con COVID-19. Método: Análisis secundario de la base de datos de la Dirección General de Epidemiología. Se incluyeron niños menores de 19 años, en quienes se confirmó SARS-CoV-2 mediante RT-PCR. Resultados: Se incluyeron 1443 niños. La mediana de edad fue de ocho años; 3.3 % ingresó a la unidad de cuidados intensivos, 1.8 % requirió ventilación mecánica asistida y la mortalidad fue de 1.9 %. En los modelos multivariados, el desarrollo de neumonía constituyó el principal factor de riesgo de mortalidad, con razón de momios (RM) de 6.45 (IC 95 % 1.99, 20.89); los pacientes que requirieron intubación tuvieron RM de 8.75 (IC 95 % 3.23, 23.7). Conclusiones: Los niños con COVID 19 tienen alta mortalidad en México, por lo que en ellos se debe procurar evitar la neumonía, especialmente en los menores de cuatro años, con riesgo cardiovascular o inmunosupresión.

Abstract Introduction: Most children affected by SARS-CoV-2 are reported to be asymptomatic, and COVID-19-related mortality in them is low; in Mexico, there is a lack of information on the subject in this population group. Objective: To assess the risk factors associated with mortality in Mexican children with COVID-19. Method: Secondary analysis of the General Directorate of Epidemiology database. Children younger than 19 years, in whom SARS-CoV-2 infection was confirmed by RT-PCR, were included. Results: 1443 children were included. Median age was eight years; 3.3 % were admitted to the intensive care unit, 1.8 % required assisted mechanical ventilation, and mortality was 1.9 %. In multivariate models, the development of pneumonia was the main risk factor for mortality, with an odds ratio (OR) of 6.45 (95 % CI 1.99, 20.89); patients who required intubation had an OR of 8.75 (95 % CI 3.23, 23.7). Conclusions: Children with COVID-19 exhibit high mortality in Mexico, and avoiding pneumonia should therefore be tried in them, especially in children younger than four years with cardiovascular risk or immunosuppression.

El daño colateral de la atención de la pandemia de COVID-19.

Throughout history, pandemics have had a major impact on humanity. The measures used to combat them cause collateral damage. During the COVID-19 pandemic, the actions taken to reduce the exposure, the number of infections, and the case fatality rate focus on reducing mortality, however, the collapse of the health system can cause an even greater number of deaths. At the same time, both medical personnel and patients are affected by the economic slowdown and the "effect of negativity". In this review article the different tools available for pandemic control, their development in a historical context, and how they may impact risk stratification for vulnerable patients (elderly, patients with chronic degenerative and oncological diseases) were analyzed.

A lo largo de la historia, las pandemias han tenido un gran impacto para la humanidad. Las medidas utilizadas para combatirlas causan daño colateral. En la pandemia por COVID-19, las acciones generadas para disminuir la exposición, el número de contagios y la tasa de letalidad conllevan un enfoque en la reducción de la mortalidad, sin embargo el colapso del sistema de salud puede provocar un número aún mayor de muertos. A su vez, tanto el personal médico como los pacientes se ven afectados por la desaceleración económica y el "efecto de la negatividad". En este artículo de revisión se analizaron las diferentes herramientas para el control de la pandemia, su desarrollo en un contexto histórico y como impactan en la estratificación del riesgo para pacientes vulnerables (ancianos, pacientes con enfermedades crónico degenerativas y oncológicos).

Deamidated Human Triosephosphate Isomerase is a Promising Druggable Target

Therapeutic strategies for the treatment of any severe disease are based on the discovery and validation of druggable targets. The human genome encodes only 600-1500 targets for small-molecule drugs, but posttranslational modifications lead to a considerably larger druggable proteome. The spontaneous conversion of asparagine (Asn) residues to aspartic acid or isoaspartic acid is a frequent modification in proteins as part of the process called deamidation. Triosephosphate isomerase (TIM) is a glycolytic enzyme whose deamidation has been thoroughly studied, but the prospects of exploiting this phenomenon for drug design remain poorly understood. The purpose of this study is to demonstrate the properties of deamidated human TIM (HsTIM) as a selective molecular target. Using in silico prediction, in vitro analyses, and a bacterial model lacking the tim gene, this study analyzed the structural and functional differences between deamidated and nondeamidated HsTIM, which account for the efficacy of this protein as a druggable target. The highly increased permeability and loss of noncovalent interactions of deamidated TIM were found to play a central role in the process of selective enzyme inactivation and methylglyoxal production. This study elucidates the properties of deamidated HsTIM regarding its selective inhibition by thiol-reactive drugs and how these drugs can contribute to the development of cell-specific therapeutic strategies for a variety of diseases, such as COVID-19 and cancer.