Differential expression of biomarkers in saliva related to SARS-CoV-2 infection in patients with mild, moderate and severe COVID-19.

BACKGROUND: Severe COVID-19 is a disease characterized by profound dysregulation of the innate immune system. There is a need to identify highly reliable prognostic biomarkers that can be rapidly assessed in body fluids for early identification of patients at higher risk for hospitalization and/or death. This study aimed to assess whether differential gene expression of immune response molecules and cellular enzymes, detected in saliva samples of COVID-19 patients, occurs according to disease severity staging. METHODS: In this cross-sectional study, subjects with a COVID-19 diagnosis were classified as having mild, moderate, or severe disease based on clinical features. Transcripts of genes encoding 6 biomarkers, IL-1ß, IL-6, IL-10, C-reactive protein, IDO1 and ACE2, were measured by RT‒qPCR in saliva samples of patients and COVID-19-free individuals. RESULTS: The gene expression levels of all 6 biomarkers in saliva were significantly increased in severe disease patients compared to mild/moderate disease patients and healthy controls. A significant strong inverse relationship between oxemia and the level of expression of the 6 biomarkers (Spearman's correlation coefficient between -0.692 and -0.757; p < 0.001) was found. CONCLUSIONS: Biomarker gene expression determined in saliva samples still needs to be validated as a potentially valuable predictor of severe clinical outcomes early at the onset of COVID-19 symptoms.

A parallel-group, multicenter randomized, double-blinded, placebo-controlled, phase 2/3, clinical trial to test the efficacy of pyridostigmine bromide at low doses to reduce mortality or invasive mechanical ventilation in adults with severe SARS-CoV-2 infection: the Pyridostigmine In Severe COvid-19 (PISCO) trial protocol.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, the causative agent of coronavirus disease 2019 (COVID-19), may lead to severe systemic inflammatory response, pulmonary damage, and even acute respiratory distress syndrome (ARDS). This in turn may result in respiratory failure and in death. Experimentally, acetylcholine (ACh) modulates the acute inflammatory response, a neuro-immune mechanism known as the inflammatory reflex. Recent clinical evidence suggest that electrical and chemical stimulation of the inflammatory reflex may reduce the burden of inflammation in chronic inflammatory diseases. Pyridostigmine (PDG), an ACh-esterase inhibitor (i-ACh-e), increases the half-life of endogenous ACh, therefore mimicking the inflammatory reflex. This clinical trial is aimed at evaluating if add-on of PDG leads to a decrease of invasive mechanical ventilation and death among patients with severe COVID-19. METHODS: A parallel-group, multicenter, randomized, double-blinded, placebo-controlled, phase 2/3 clinical trial to test the efficacy of pyridostigmine bromide 60 mg/day P.O. to reduce the need for invasive mechanical ventilation and mortality in hospitalized patients with severe COVID-19. DISCUSSION: This study will provide preliminary evidence of whether or not -by decreasing systemic inflammation- add-on PDG can improve clinical outcomes in patients with severe COVID-19. TRIAL REGISTRATION: ClinicalTrials.gov NCT04343963 (registered on April 14, 2020).

Antiretroviral purchasing and prescription practices in Mexico: constraints, challenges and opportunities.

OBJECTIVE: This study examines the antiretroviral (ARV) market characteristics for drugs procured and prescribed to Mexico's Social Protection System in Health beneficiaries between 2008 and 2013, and compares them with international data. MATERIALS AND METHODS: Procurement information from the National Center for the Prevention and the Control of HIV/AIDS was analyzed to estimate volumes and prices of key ARV. Annual costs were compared with data from the World Health Organization's Global Price Reporting Mechanism for similar countries. Finally, regimens reported in the ARV Drug Management, Logistics and Surveillance System database were reviewed to identify prescription trends and model ARV expenditures until 2018. RESULTS: Results show that the first-line ARV market is concentrated among a small number of patented treatments, in which prescription is clinically adequate, but which prices are higher than those paid by similar countries. The current set of legal and structural options available to policy makers to bring prices down is extremely limited. CONCLUSIONS: Different negotiation policies were not successful to decrease ARV high prices in the public health market. The closed list approach had a good impact on prescription quality but was ineffective in reducing prices. The Coordinating Commission for Negotiating the Price of Medicines and other Health Supplies also failed to obtain adequate prices. To maximize purchase efficiency, policy makers should focus on finding long-term legal and political safeguards to counter the high prices imposed by pharmaceutical companies.

Interim safety and immunogenicity results from an NDV-based COVID-19 vaccine phase I trial in Mexico.

There is still a need for safe, efficient, and low-cost coronavirus disease 2019 (COVID-19) vaccines that can stop transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here we evaluated a vaccine candidate based on a live recombinant Newcastle disease virus (NDV) that expresses a stable version of the spike protein in infected cells as well as on the surface of the viral particle (AVX/COVID-12-HEXAPRO, also known as NDV-HXP-S). This vaccine candidate can be grown in embryonated eggs at a low cost, similar to influenza virus vaccines, and it can also be administered intranasally, potentially to induce mucosal immunity. We evaluated this vaccine candidate in prime-boost regimens via intramuscular, intranasal, or intranasal followed by intramuscular routes in an open-label non-randomized non-placebo-controlled phase I clinical trial in Mexico in 91 volunteers. The primary objective of the trial was to assess vaccine safety, and the secondary objective was to determine the immunogenicity of the different vaccine regimens. In the interim analysis reported here, the vaccine was found to be safe, and the higher doses tested were found to be immunogenic when given intramuscularly or intranasally followed by intramuscular administration, providing the basis for further clinical development of the vaccine candidate. The study is registered under ClinicalTrials.gov identifier NCT04871737.

Safety and immunogenicity of a live recombinant Newcastle disease virus-based COVID-19 vaccine (Patria) administered via the intramuscular or intranasal route: Interim results of a non-randomized open label phase I trial in Mexico.

There is still a need for safe, efficient and low-cost coronavirus disease 2019 (COVID-19) vaccines that can stop transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here we evaluated a vaccine candidate based on a live recombinant Newcastle disease virus (NDV) that expresses a stable version of the spike protein in infected cells as well as on the surface of the viral particle (AVX/COVID-12-HEXAPRO, also known as NDV-HXP-S). This vaccine candidate can be grown in embryonated eggs at low cost similar to influenza virus vaccines and it can also be administered intranasally, potentially to induce mucosal immunity. We evaluated this vaccine candidate in prime-boost regimens via intramuscular, intranasal, or intranasal followed by intramuscular routes in an open label non-randomized non-placebo-controlled phase I clinical trial in Mexico in 91 volunteers. The primary objective of the trial was to assess vaccine safety and the secondary objective was to determine the immunogenicity of the different vaccine regimens. In the interim analysis reported here, the vaccine was found to be safe and the higher doses tested were found to be immunogenic when given intramuscularly or intranasally followed by intramuscular administration, providing the basis for further clinical development of the vaccine candidate. The study is registered under ClinicalTrials.gov identifier NCT04871737. Funding was provided by Avimex and CONACYT.

Integrated food chain surveillance system for Salmonella spp. in Mexico.

Few developing countries have foodborne pathogen surveillance systems, and none of these integrates data from humans, food, and animals. We describe the implementation of a 4-state, integrated food chain surveillance system (IFCS) for Salmonella spp. in Mexico. Significant findings were 1) high rates of meat contamination (21.3%-36.4%), 2) high rates of ceftriaxone-resistant S. Typhimurium in chicken, ill humans, and swine (77.3%, 66.3%, and 40.4% of S. Typhimurium T isolates, respectively), and 3) the emergence of ciprofloxacin resistance in S. Heidelberg (10.4%) and S. Typhimurium (1.7%) from swine. A strong association between Salmonella spp. contamination in beef and asymptomatic Salmonella spp. infection was only observed in the state with the lowest poverty level (Pearson r = 0.91, p<0.001). Pulsed-field gel electrophoresis analysis of 311 S. Typhimurium isolates showed 14 clusters with 102 human, retail meat, and food-animal isolates with indistinguishable patterns. An IFCS is technically and economically feasible in developing countries and can effectively identify major public health priorities.

Antimicrobial susceptibility patterns of Streptococcus pneumoniae in Mexico.

The susceptibility to 14 beta-lactam and non-beta-lactam antimicrobial agents was evaluated for Streptococcus pneumoniae from patients with community-acquired respiratory infections in a Mexican medical center. Three hundred fifteen pneumococcal isolates obtained from patients between 1995 and 2001 were tested by the broth microdilution test. Fifty-two percent of the isolates were nonsusceptible to penicillin (minimal inhibitory concentration, >0.06 microg/mL). Penicillin-nonsusceptible isolates were more likely to exhibit resistance to cephalosporins, macrolides, ciprofloxacin, trimethoprim/sulfamethoxazole, chloramphenicol, and tetracycline when compared to penicillin-susceptible isolates. Ninety-three percent of the penicillin-nonsusceptible isolates were resistant to at least one other class of antimicrobials, in contrast to only 47% of the penicillin-susceptible strains (p < 0.0001). More than 90% of the tested isolates were susceptible to amoxicillin/clavulanate, ceftriaxone, levofloxacin, and gatifloxacin. Reduced susceptibility to penicillin was considered to be a reliable marker for the higher probability of multidrug resistance, thus requiring in vitro tests to guide chemotherapy or the choices of parenteral extended spectrum cephalosporins or newer respiratory quinolones.

Antiretroviral purchasing and prescription practices in Mexico: constraints, challenges and opportunities / Compras y prescripciones de medicamentos antirretrovirales en México: restricciones, retos y oportunidades

Objective. This study examines the antiretroviral (ARV) market characteristics for drugs procured and prescribed to Mexico's Social Protection System in Health beneficiaries between 2008 and 2013, and compares them with international data. Materials and methods. Procurement information from the National Center for the Prevention and the Control of HIV/AIDS was analyzed to estimate volumes and prices of key ARV. Annual costs were compared with data from the World Health Organization's Global Price Reporting Mechanism for similar countries. Finally, regimens reported in the ARV Drug Management, Logistics and Surveillance System database were reviewed to identify prescription trends and model ARV expenditures until 2018. Results. Results show that the first-line ARV market is concentrated among a small number of patented treatments, in which prescription is clinically adequate, but which prices are higher than those paid by similar countries. The current set of legal and structural options available to policy makers to bring prices down is extremely limited. Conclusions. Different negotiation policies were not successful to decrease ARV high prices in the public health market. The closed list approach had a good impact on prescription quality but was ineffective in reducing prices. The Coordinating Commission for Negotiating the Price of Medicines and other Health Supplies also failed to obtain adequate prices. To maximize purchase efficiency, policy makers should focus on finding long-term legal and political safeguards to counter the high prices imposed by pharmaceutical companies.

Objetivo. Este estudio analiza el mercado de los medicamentos antiretrovirales (ARV) adquiridos y prescritos a los beneficiarios del Seguro Popular entre 2008 y 2013, en México, comparándolo con información internacional. Material y métodos. Se analiza información sobre la compra de medicamentos por parte del Centro para la Prevención y el Control del VIH y el Sida (Censida) para estimar precios y volúmenes de compra de los principales ARV. Los costos anuales de tratamiento estimados fueron comparados con información del Global Price Reporting Mechanism (GPRM) de la Organización Mundial de la Salud, para países similares. Finalmente se revisaron los esquemas reportados en el Sistema de Administración, Logística y Vigilancia de ARV para identificar tendencias y proyectar el gasto en ARV hasta 2018. Resultados. El mercado mexicano de ARV está concentrado en pocos esquemas de primera línea y, aunque la prescripción es clínicamente adecuada, los precios son más altos que en otros países similares. El conjunto actual de opciones legales y estructurales disponibles para los formuladores de políticas para reducir los precios es muy limitado. Conclusiones. Las políticas de negociación han sido poco exitosas para disminuir los precios de los ARV en México. La Coordinating Commission for Negotiating the Price of Medicines and other Health Supplies y la integración de las guías de tratamiento han tenido impacto significativo en la calidad de la prescripción, pero moderado en la reducción de precios. Por ello es necesario buscar garantías jurídicas y políticas a largo plazo para hacer frente a los altos precios de los ARV.

Phenotypic differences between BCG vaccines at the proteome level.

To contribute to Mycobacterium bovis BCG characterization, two substrains were analyzed using two-dimensional gel electrophoresis (2D-PAGE) and mass spectrometry (MS), based on their protective efficacy in a pulmonary-tuberculosis mouse model. Cell-fraction proteins of BCG Denmark and Phipps substrains were separated into approximately 500 spots in 2D-PAGE. The proteomes were similar in protein number, and isoelectric point (pI) and molecular mass (MM) distribution. Statistical analysis, resulted in 72 spots with no change, and 168 and 90 unique for BCG Phipps or Denmark, respectively. Two hundred and fourteen spots showed changes in intensity of >1-fold, 138 of Denmark, and 76 of Phipps. Seventeen spots were selected for MS-based identification (13 from Phipps and 4 from Denmark), including unique, as well as proteins with changes in intensity. The proteins identified participate in virulence, detoxification, adaptation, lipid metabolism, information pathways, cell wall and cell processes, intermediary metabolism and respiration, or still hypotheticals. Our findings contribute to phenotype characterization of BCG substrains and provide new elements to consider for the design of diagnostic tools, drug targets and a new vaccine against tuberculosis based upon protein expression through quantitative statistical analysis.

Mycobacterium bovis BCG substrains confer different levels of protection against Mycobacterium tuberculosis infection in a BALB/c model of progressive pulmonary tuberculosis.

Mycobacterium bovis BCG is the only available vaccine against tuberculosis. Reasons for why diverse BCG substrains induce different levels of protection in clinical trials remain unclear. The aim of this study was to compare the effectiveness of 10 BCG substrains in a mouse model of pulmonary tuberculosis. BALB/c mice were subcutaneously vaccinated and 2 months later were challenged with Mycobacterium tuberculosis H37Rv by intratracheal injection. Two and 4 months after challenge, delayed-type hypersensitivity (DTH) response, lung tissue affected by pneumonia, CFU, T-cell counts, and cytokine expression (interleukin-2 [IL-2], IL-4, IL-10, and gamma interferon) were determined. A differential protective effect of the diverse BCG substrains was found. BCG Phipps led to the largest and most persistent reduction of CFU counts and of the area of pneumonia at 2 and 4 months after challenge. This protection was accompanied by reduced IL-10-producing T cells. Contemporary BCG substrains induce a wide range of protection in this animal model. These data can help in the selection of the best vaccine for human immunization and for the development of novel recombinant BCG-based vaccine.

Longitudinal study of microbial diversity and seasonality in the Mexico City metropolitan area water supply system.

In the Mexico City metropolitan area (MCMA), 70% of the water for 18 million inhabitants is derived from the Basin of Mexico regional aquifer. To provide an overview of the quality of the groundwater, a longitudinal study was conducted, in which 30 sites were randomly selected from 1,575 registered extraction wells. Samples were taken before and after chlorine disinfection during both the rainy and dry seasons (2000-2001). Microbiological parameters (total coliforms, fecal coliforms, streptococci, and Vibrio spp.), the presence of Helicobacter pylori, and physicochemical parameters, including the amount of trihalomethanes (THMs), were determined. Although microorganisms and inorganic and organic compounds were evident, they did not exceed current permissible limits. Chlorine levels were low, and the bacterial counts were not affected by chlorine disinfection. Eighty-four bacterial species from nine genera normally associated with fecal contamination were identified in water samples. H. pylori was detected in at least 10% of the studied samples. About 40% of the samples surpassed the THM concentration allowed by Mexican and U.S. regulations, with levels of chloroform being high. The quality of the water distributed to the MCMA varied between the rainy and dry seasons, with higher levels of pH, nitrates, chloroform, bromodichloromethane, total organic carbon, and fecal streptococci during the dry season. This study showed that the groundwater distribution system is susceptible to contamination and that there is a need for a strict, year-round disinfection strategy to ensure adequate drinking-water quality. This situation in one of the world's megacities may reflect what is happening in large urban centers in developing countries which rely on a groundwater supply.

Rotavirus diarrhea severity is related to the VP4 type in Mexican children.

This report is of a community-based case control study to assess whether the severity of acute diarrhea by rotavirus (RV) in young children is associated with a particular VP7 (G) or VP4 (P) RV serotype. Five hundred twenty children younger than 2 years of age with diarrhea lasting less than 3 days were age and gender matched with 520 children with no diarrhea. The G and P serotypes were determined with specific monoclonal antibodies, and the VP4 serotype specificity in a subgroup was confirmed by genotyping. Infection with a G3 serotype led to a higher risk of diarrhea than infection with a G1 serotype. Infection with a G3-nontypeable-P serotype was associated with more severe gastroenteritis than infection with a G3 (or G1) P1A[8] serotype. A child with diarrhea-associated dehydration was almost five times more likely to be infected with a G3-nontypeable-P serotype than a child without dehydration (P < 0.001). Moreover, the two predominant monotypes within serotype P1A[8] had significantly different clinical manifestations. In this study, the severity of RV-associated diarrhea was related to different P serotypes rather than to G serotypes. The relationship between serotype and clinical outcomes seems to be complex and to vary among different geographic areas.

Utilidad de la telerradiografía de tórax en pacientes hospitalizados / Usefulness of the chest roentgenogram in hospitalized patients

Objetivo. Evaluar la utilidad de la telerradiografía de tórax (TRT) en enfermos hospitalizados, en términos de su influencia en el plan diagnóstico y terapéutico anticipado por el clínico. Diseño. Encuesta a un grupo de médicos residentes en medicina interna encargados de los sectores de hospitalización del Instituto Nacional de la Nutrición Salvador Zubirán. Se aplicaron dos tipos de cuestionarios: uno al momento de la solicitud de una TRT (motivo de la solicitud, la probabilidad de un hallazgo anormal y su decisión terapéutica) y otro, una vez conocido el resultado de la radiografía, averiguando lo inseperado del hallazgo radiológico y su influencia en la terapéutica. Se analizaron las respuestas a estos cuestionarios a la luz de la interpretación de la placa por un radiólogo. Resultados. Se analizaron 100 encuestas. La TRT descartó la sospecha anticipada por el médico de aparición de un nuevo evento pulmonar (55 por ciento de las veces) o de una mala evolución de una enfermedad pulmonar ya conocida (en el 50 por ciento). En cambio, la TRT corroboró la ausencia de un nuevo evento pulmonar, de una mejor o estable evolución de una enfermedad pulmonar ya conocida, anticipados por el médico, en el 92 por ciento de los casos. La TRT determinó un cambio en el plan terapéutico anticipado por el clínico en el 61 por ciento de las veces. Conclusión. La TRT en pacientes hospitalizados es de utilidad práctica al médico al evitar sobrediagnósticos clínicos y el consecuente tratamiento innecesario de sus enfermos

Resistencia a antimicrobianos de helicobacter pylori en un centro de referencia infectológico / Antimicrobial resistance in isolates of Helicobacter pylori in a referral center

Objetivo. Estimar la frecuencia de aislados clínicos de H. pylori resistentes a seis antibióticos de uso común en su erradicación. Diseño. Estudio observacional transversal. Lugar. Institución hospitalaria de referencia de tercer nivel. Participantes. 31 aislamientos de igual número de enfermos con gastritis antral crónica, obtenidos de biopsias de mucosa gástrica. Desenlace principal. Se midió la concentración mínima anhibitoria (CMI) de metronidazol, tetraciclina, doxiciclina, ampicilina, amoxicilina y de subcitrato de bismuto mediante la técnia de dilución en placas de agar. Resultados. Todos los aislados mostraron ser sensibles a tetraciclina, doxiciclina, ampicilina y a amoxicilina; sólo 46 por ciento y 55 por ciento fueron inhibidos a concentraciones menores a 8 µg/mL y a 16 µg/mL de metronidazol, respectivamente. Todos los aislamientos fueron inhibidos a una concentración =128 µg/mL de la sal de bismuto. Se observó un incremento del 50 por ciento en el porcentaje de aislados resistentes a metronidazol (resistencia definida como una CMI = 8µg/mL) al comparar los de 1988 con los de 1992. Conclusión. Se necesitan estudios a futuro que evalúen cuál esquema de antibioticoterapia ofrece un mejor índice costo/beneficio en el tratamiento de la úlcera péptica por H. pylori en nuestro medio

Epidemiological and biological characteristics of Methicillin-Resistant staphylococcal infections in a Mexican Hospital

Background. Methicillin-resistant Staphylococcus aureus (MRSA) has spread worldwide since 1960. However, there is little information concerning methicillinresistant coagulase-negative staphylococci (MRCNS) infections. Methods. In order to study the clinical and epidemiological characteristics of methicillinresistant staphylococci (MRS) infections and to determine the relationship between MRS and both synergistic hemolysis (SH) and slime production (SP), a laboratory-based survey and non-matched case-control study were carried out at a tertiary-care center in Mexico City. In regard to patients, from May 1991 to October 1992, 46 cases of MRS infection and 86 patients (controls) infected by methicillin-susceptible staphylococci (MSS) were included. Clinical and epidemiologic variables were analyzed. The isolates were identified and tested for antimicrobial susceptibility by standard method. An MIC of oxacillin = 8 µg/mL was defined as an MRS. Results. During the study. 94 nosocomial staphyloccocal infections were diagnosed: S. aureus, 35 and CNS, 59; 43 (45.7 percent) by MRS (rate of MRS infections was 1.12 per 100 inpatients); 2 MRSA; 41 MRCNS, and only 19 were symptomatic. Three infections were community-acquired, including one MRSA and two MRCNS. After multivariate analysis, the significant risk factors were previous antimicrobial therapy (p= 0.013) and catheterelated (p= 0.009) and urinary-tract source (p = 0.0001). Forty-nine percent of MRS showed SH while only 15 percent of MSS (p < 0.001) showed SH, especially in 10/10 MR-S. hemolyticus. additionally, 48 percent of MRCNS showed SP, as did 18 percent of MSCNS (p = 0.019), particularly in 15/20 MR-S. epidermis. Of all MRS isolates, 38 percent showed a homogeneous phenotype, a trait associated with multi-durg resistance (p < 0.01) and SH (p< 0.001). Conclusions. CNS predominanted as the cause of MRS infections in our setting. The homogeneous phenotype was associated with SH and multi-drug resistance

Patrones de susceptibilidad a 19 antimicrobianos de germenes aislados de hemocultivos en un hospital de referencia de la ciudad de México / Sensitivity patterns to 19 antimicrobial agents of bacterial strains isolated from blood cultures in a referencie hospital of Mexico city

Se midió la sensiblidad a 19 antimicrobianos de 598 cepas bacterianas obtenidas de cultivos sanguíneos durante un lapso de 23 meses (diciembre de 1982-octubre de 1984), correspondiendo cada una de estas cepas a diferentes pacientes vistos en el Instituto Nacional de la Nutrición Salvador Zubirán. Mediante la determinación de las concentraciones mínimas inhibitorias, efectuadas por el método de icrominación el placa, se reportan los patrones de resistencia en los años de 1983 y 1984 de los gérmenes estudiados. Los aminoglucósidos (amikacina, gentamicina y tobramicina) mostraron tener buena actividad bacteriostática sobre la mayoría de las cepas probadas. En contraste, los porcentajes de cepas sensibles a la mayoría de los ß-lactámicos probados fueron menores, sobre todo los de Pseudomonas aeruginosa y Serratia-sp. La cefalotina y la dicloxacilina mostraron buena actividad sobre los cocos gram positivos. El aztreonam y la norfloxacina inhibieron casi todas las cepas probadas. Comparando los porcentajes de cepas resistentes en el período de 1983 con el de 1984, no hubo cambios estadísticamente significativos, excepto para un incremento de 100% de cepas resistentes de Pseudomonas a la carbenicilina y de un aumento de cepas resistentes de Klebsiella a la tobramicina. Es necesario continuar esta vigilancia epidemiológica de los patrones de resistencia de las cepas aisladas de hemocultivos por tratarse de una información indispensable para el manejo empírico de antibióticos en los pacientes con bacteremia