Resistance phenotype and virulence potential of Leclercia adecarboxylata strains isolated from different sources.

Introduction. Leclercia adecarboxylata is a member of Enterobacterales, often considered an opportunistic pathogen. Recent reports have highlighted L. adecarboxylata as an emerging pathogen harbouring virulence and resistance determinants.Gap statement. Little information exists on virulence and resistance determinants in L. adecarboxylata strains isolated from environmental, food, and clinical samples.Aim. To determine the presence of resistance and virulence determinants and plasmid features in L. adecarboxylata strains isolated from environmental, food, and clinical samples, as well as their phylogenetic relationship.Results. All strains tested showed resistance to ß-lactams and quinolones but were sensitive to aminoglycosides and nitrofurans. However, even though fosfomycin resistance is considered a characteristic trait of L. adecarboxylata, the resistance phenotype was only observed in 50 % of the strains; bla TEM was the most prevalent BLEE gene (70 %), while the quinolone qnrB gene was observed in 60 % of the strains. Virulence genes were differentially observed in the strains, with adhesion-related genes being the most abundant, followed by toxin genes. Finally, all strains carried one to seven plasmid bands ranging from 7 to 125 kbps and harboured several plasmid addiction systems, such as ParDE, VagCD, and CcdAB in 80 % of the strains.Conclusions. L. adecarboxylata is an important emerging pathogen that may harbour resistance and virulence genes. Additionally, it has mobilizable genetic elements that may contribute to the dissemination of genetic determinants to other bacterial genera.

Genomic Analysis of Early SARS-CoV-2 Variants Introduced in Mexico.

The coronavirus disease 2019 (COVID-19) pandemic has affected most countries in the world. Studying the evolution and transmission patterns in different countries is crucial to enabling implementation of effective strategies for disease control and prevention. In this work, we present the full genome sequence for 17 SARS-CoV-2 isolates corresponding to the earliest sampled cases in Mexico. Global and local phylogenomics, coupled with mutational analysis, consistently revealed that these viral sequences are distributed within 2 known lineages, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) lineage A/G, containing mostly sequences from North America, and lineage B/S, containing mainly sequences from Europe. Based on the exposure history of the cases and on the phylogenomic analysis, we characterized 14 independent introduction events. Additionally, three cases with no travel history were identified. We found evidence that two of these cases represented local transmission cases occurring in Mexico during mid-March 2020, denoting the earliest events described for the country. Within this local transmission cluster, we also identified an H49Y amino acid change in the Spike protein. This mutation represents a homoplasy occurring independently through time and space and may function as a molecular marker to follow any further spread of these viral variants throughout the country. Our results provide a general picture of the SARS-CoV-2 variants introduced at the beginning of the outbreak in Mexico, setting the foundation for future surveillance efforts.IMPORTANCE Understanding the introduction, spread, and establishment of SARS-CoV-2 within distinct human populations as well as the evolution of the pandemics is crucial to implement effective control strategies. In this work, we report that the initial virus strains introduced in Mexico came from Europe and the United States and that the virus was circulating locally in the country as early as mid-March. We also found evidence for early local transmission of strains with a H49Y mutation in the Spike protein, which could be further used as a molecular marker to follow viral spread within the country and the region.

Association between bacterial homoplastic variants and radiological pathology in tuberculosis.

BACKGROUND: Understanding how pathogen genetic factors contribute to pathology in TB could enable tailored treatments to the most pathogenic and infectious strains. New strategies are needed to control drug-resistant TB, which requires longer and costlier treatment. We hypothesised that the severity of radiological pathology on the chest radiograph in TB disease was associated with variants arising independently, multiple times (homoplasies) in the Mycobacterium tuberculosis genome. METHODS: We performed whole genome sequencing (Illumina HiSeq2000 platform) on M. tuberculosis isolates from 103 patients with drug-resistant TB in Lima between 2010 and 2013. Variables including age, sex, HIV status, previous TB disease and the percentage of lung involvement on the pretreatment chest radiograph were collected from health posts of the national TB programme. Genomic variants were identified using standard pipelines. RESULTS: Two mutations were significantly associated with more widespread radiological pathology in a multivariable regression model controlling for confounding variables (Rv2828c.141, RR 1.3, 95% CI 1.21 to 1.39, p<0.01; rpoC.1040 95% CI 1.77 to 2.16, RR 1.9, p<0.01). The rpoB.450 mutation was associated with less extensive radiological pathology (RR 0.81, 95% CI 0.69 to 0.94, p=0.03), suggestive of a bacterial fitness cost for this mutation in vivo. Patients with a previous episode of TB disease and those between 10 and 30 years of age also had significantly increased radiological pathology. CONCLUSIONS: This study is the first to compare the M. tuberculosis genome to radiological pathology on the chest radiograph. We identified two variants significantly positively associated with more widespread radiological pathology and one with reduced pathology. Prospective studies are warranted to determine whether mutations associated with increased pathology also predict the spread of drug-resistant TB.

Presence of chelonid herpesvirus 5 (ChHV5) in sea turtles in northern Sinaloa, Mexico.

The presence of fibropapilloma and its associated chelonid herpesvirus 5 (ChHV5) was assessed in 82 wild sea turtles. Olive ridley turtles Lepidochelys olivacea (n = 58) were caught in the pelagic Area of Marine Influence (AMI) (off the coast of Guasave, Sinaloa), and black turtles Chelonia mydas agassizii (n = 24) were captured in the Navachiste Lagoon System. The apparent physical condition was evaluated as 'good' or 'poor' by physical examination. The population structure and general health status was determined by condition index, hematocrit and total plasma protein. Detection of ChHV5 from skin samples was done by PCR. The overall physical condition of black turtles was good and all the individuals were tumor-free. Likewise, the physical condition of most olive ridley turtles was good, except for 10 individuals with poor condition. Of these, 4 had fibropapilloma-like tumors. PCR analyses showed that 3 tumors were ChHV5-positive. The DNA sequence showed 96% identity with ChHV5. All other skin samples from black or olive ridley turtles were ChHV5-negative. This is the first report of fibropapillomatosis-ChHV5 in foraging grounds off northern Sinaloa. The virus was present in a small proportion of L. olivacea individuals, a free-ranging species. It is suggested that infected turtles acquired the virus at a different location somewhere during their development before arriving in the AMI zone. This finding makes the case for setting up a health monitoring program for turtle populations in the area, enforcing sanitary measures to reduce the spread of the pathogen.

Modification of HIF-1α, NF-aκB, IGFBP-3, VEGF and adiponectin in diabetic foot ulcers treated with hyperbaric oxygen.

Introduction: Diabetic foot ulcers are a frequent complication of diabetes and the first cause of non-traumatic lower limb amputation. They affect quality of life, restrict social productivity and generate a high economic burden for health care systems. Hyperbaric oxygen (HBO2) therapy is an adjunctive treatment option because it improves wound healing in the short term. However, its ability to modulate the pro- and anti-inflammatory balance and the hypoxic cell response in the clinical setting has not been fully described. Objective: To determine modifications in HIF-1α, NF-κB, IGFBP-3, and VEGF expression in wounds as well as circulating inflammatory cytokines in patients with diabetic foot ulcers subjected to HBO2. Materials and methods: We studied 17 ambulatory patients and one hospitalized patient with diabetic foot ulcers classified as Grade 3 or 4 according to the Wagner scale. All underwent HBO2 therapy. Tissue expression of HIF-1α, NF-κB, IGFBP-3, and VEGF was determined by immunohistochemistry. Plasma levels of adiponectin, IL-6, IFN-γ, IL-10 and IL-4 were measured by ELISA and chemiluminescence. Fibrosis and angiogenesis were determined by Masson's trichrome staining. Results: Ulcers in all patients healed after one month of HBO2, and none presented relapses at the one-year follow-up. At the beginning of treatment, HIF-1α and NF-κB expression was observed mainly in the nucleus, whereas these proteins were localized in the cytoplasm at the end of HBO2. There were significant modifications in VEGF expression after therapy, an increase in the plasma level of proinflammatory IL-6, and a decrease in that of IFN-γ. IGFBP-3 expression and plasma levels of adiponectin were increased at the end of HBO2. Increases in fibrosis and angiogenesis were also observed. Conclusion: These results suggest that adjuvant HBO2 modifies the proinflammatory balance related to the cellular response to hypoxia.

Differential Detection of Enterovirus and Herpes Simplex Virus in Cerebrospinal Fluid by Real-Time RT-PCR.

BACKGROUND: Enterovirus (EV) and herpes simplex virus 1 and 2 (HSV1 and HSV2) are the main etiologic agents of central nervous system infections. Early laboratory confirmation of these infections is performed by viral culture of the cerebrospinal fluid (CSF), or the detection of specific antibodies in serum (e.g., HSV). The sensitivity of viral culture ranges from 65 to 75%, with a recovery time varying from 3 to 10 days. Serological tests are faster and easy to carry out, but they exhibit cross-reactivity between HSV1 and HSV2. Although molecular techniques are more sensitive (sensitivity >95%), they are more expensive and highly susceptible to cross-contamination. METHODS: A real-time RT-PCR for the detection of EV, HSV1, and HSV2 was compared with end-point nested PCR. RESULTS: We tested 87 CSF samples of patients with a clinical diagnosis of viral meningitis or encephalitis. Fourteen samples were found to be positive by RT-PCR, but only 8 were positive by end-point PCR. The RT-PCR showed a specificity range of 94-100%, the negative predictive value was 100%, and the positive predictive value was 62, 100, and 28% for HSV1, HSV2, and EV, respectively. CONCLUSION: Real-time RT-PCR detected EV, HSV1, and HSV2 with a higher sensitivity and specificity than end-point nested RT-PCR.

Multidrug-Resistant Tuberculosis Treatment Outcomes in Relation to Treatment and Initial Versus Acquired Second-Line Drug Resistance.

BACKGROUND: Resistance to second-line drugs develops during treatment of multidrug-resistant (MDR) tuberculosis, but the impact on treatment outcome has not been determined. METHODS: Patients with MDR tuberculosis starting second-line drug treatment were enrolled in a prospective cohort study. Sputum cultures were analyzed at a central reference laboratory. We compared subjects with successful and poor treatment outcomes in terms of (1) initial and acquired resistance to fluoroquinolones and second-line injectable drugs (SLIs) and (2) treatment regimens. RESULTS: Of 1244 patients with MDR tuberculosis, 973 (78.2%) had known outcomes and 232 (18.6%) were lost to follow-up. Among those with known outcomes, treatment succeeded in 85.8% with plain MDR tuberculosis, 69.7% with initial resistance to either a fluoroquinolone or an SLI, 37.5% with acquired resistance to a fluoroquinolone or SLI, 29.3% with initial and 13.0% with acquired extensively drug-resistant tuberculosis (P < .001 for trend). In contrast, among those with known outcomes, treatment success increased stepwise from 41.6% to 92.3% as the number of drugs proven effective increased from ≤1 to ≥5 (P < .001 for trend), while acquired drug resistance decreased from 12% to 16% range, depending on the drug, down to 0%-2% (P < .001 for trend). In multivariable analysis, the adjusted odds of treatment success decreased 0.62-fold (95% confidence interval, .56-.69) for each increment in drug resistance and increased 2.1-fold (1.40-3.18) for each additional effective drug, controlling for differences between programs and patients. Specific treatment, patient, and program variables were also associated with treatment outcome. CONCLUSIONS: Increasing drug resistance was associated in a logical stepwise manner with poor treatment outcomes. Acquired resistance was worse than initial resistance to the same drugs. Increasing numbers of effective drugs, specific drugs, and specific program characteristics were associated with better outcomes and less acquired resistance.

Impact of HIV on mortality among patients treated for tuberculosis in Lima, Peru: a prospective cohort study.

BACKGROUND: Human immunodeficiency virus (HIV)-associated tuberculosis deaths have decreased worldwide over the past decade. We sought to evaluate the effect of HIV status on tuberculosis mortality among patients undergoing treatment for tuberculosis in Lima, Peru, a low HIV prevalence setting. METHODS: We conducted a prospective cohort study of patients treated for tuberculosis between 2005 and 2008 in two adjacent health regions in Lima, Peru (Lima Ciudad and Lima Este). We constructed a multivariate Cox proportional hazards model to evaluate the effect of HIV status on mortality during tuberculosis treatment. RESULTS: Of 1701 participants treated for tuberculosis, 136 (8.0%) died during tuberculosis treatment. HIV-positive patients constituted 11.0% of the cohort and contributed to 34.6% of all deaths. HIV-positive patients were significantly more likely to die (25.1 vs. 5.9%, P < 0.001) and less likely to be cured (28.3 vs. 39.4%, P = 0.003). On multivariate analysis, positive HIV status (hazard ratio [HR] = 6.06; 95% confidence interval [CI], 3.96-9.27), unemployment (HR = 2.24; 95% CI, 1.55-3.25), and sputum acid-fast bacilli smear positivity (HR = 1.91; 95% CI, 1.10-3.31) were significantly associated with a higher hazard of death. CONCLUSIONS: We demonstrate that positive HIV status was a strong predictor of mortality among patients treated for tuberculosis in the early years after Peru started providing free antiretroviral therapy. As HIV diagnosis and antiretroviral therapy provision are more widely implemented for tuberculosis patients in Peru, future operational research should document the changing profile of HIV-associated tuberculosis mortality.

[Descriptive epidemiology of children with acute myeloid leukemia residing in Mexico City: a report from the Mexican Inter-Institutional Group for Identifying Childhood Leukemia Causes]. / Epidemiología descriptiva de la leucemia mieloide aguda (LMA) en niños residentes de la Ciudad de México: reporte del Grupo Mexicano Interinstitucional para la Identificación de las Causas de la Leucemia en Niños.

INTRODUCTION: Acute myeloid leukemias represent the second most common childhood leukemia subtype. In Mexico, there are few studies on descriptive epidemiology for this disease. AIMS: To report acute myeloid leukemia incidence for children less than 15 years of age in the Metropolitan Area of the Valley of Mexico for a period of five years (2010-2014) and to analyze whether there are differences in the incidence of acute myeloid leukemia by regions. MATERIAL AND METHODS: A descriptive study was conducted in nine public hospitals in Mexico City. The crude annual average incidence rate and adjusted average annual incidence rate were calculated. RESULTS: A total of 190 patients with diagnosis of de novo acute myeloid leukemia were analyzed. Male sex (57.2%) and acute myeloid leukemia-M3 subtype (25.3%) were more frequent. The adjusted average annual incidence rates for Mexico City and for the Metropolitan Area of the Valley of Mexico were 8.18 and 7.74 per million children under 15 years old, respectively. CONCLUSIONS: It seems that childhood acute myeloid leukemia incidence is increasing in Mexico City, which makes the identification of associated risk factors imperative.

Prevalence of and risk factors for resistance to second-line drugs in people with multidrug-resistant tuberculosis in eight countries: a prospective cohort study.

BACKGROUND: The prevalence of extensively drug-resistant (XDR) tuberculosis is increasing due to the expanded use of second-line drugs in people with multidrug-resistant (MDR) disease. We prospectively assessed resistance to second-line antituberculosis drugs in eight countries. METHODS: From Jan 1, 2005, to Dec 31, 2008, we enrolled consecutive adults with locally confirmed pulmonary MDR tuberculosis at the start of second-line treatment in Estonia, Latvia, Peru, Philippines, Russia, South Africa, South Korea, and Thailand. Drug-susceptibility testing for study purposes was done centrally at the Centers for Disease Control and Prevention for 11 first-line and second-line drugs. We compared the results with clinical and epidemiological data to identify risk factors for resistance to second-line drugs and XDR tuberculosis. FINDINGS: Among 1278 patients, 43·7% showed resistance to at least one second-line drug, 20·0% to at least one second-line injectable drug, and 12·9% to at least one fluoroquinolone. 6·7% of patients had XDR tuberculosis (range across study sites 0·8-15·2%). Previous treatment with second-line drugs was consistently the strongest risk factor for resistance to these drugs, which increased the risk of XDR tuberculosis by more than four times. Fluoroquinolone resistance and XDR tuberculosis were more frequent in women than in men. Unemployment, alcohol abuse, and smoking were associated with resistance to second-line injectable drugs across countries. Other risk factors differed between drugs and countries. INTERPRETATION: Previous treatment with second-line drugs is a strong, consistent risk factor for resistance to these drugs, including XDR tuberculosis. Representative drug-susceptibility results could guide in-country policies for laboratory capacity and diagnostic strategies. FUNDING: US Agency for International Development, Centers for Disease Control and Prevention, National Institutes of Health/National Institute of Allergy and Infectious Diseases, and Korean Ministry of Health and Welfare.

Genetic polymorphisms of the tumor necrosis factor and lymphotoxin alpha in type 2 diabetes.

OBJECTIVE: To investigate the frequency of the single-base change polymorphic variants identified in tumor necrosis factor (TNF) gene (-308 G/A) and lymphotoxin alpha (LTA) (+252 G/A) in patients with type 2 diabetes (T2D). METHODS: A prospective study in a Mexican-mestizo population of 51 patients with T2D and 48 healthy subjects was carried out. We took a peripheral blood sample from each individual for identification of the polymorphic genotypes by polymerase chain reaction. RESULTS: The genotype distribution in T2D was: TNF alpha homozygous 0%; TNFG/A heterozygous 20%; TNFG homozygous 80%. CONCLUSIONS: In regards to the TNF -308 G/A genotype, we found a significant difference (p = 0.012) with a bigger frequency in the group of patients. The health controls showed a higher frequency of TNF -308 G/G genotype (p = 0.034).

[Mixed analysis of the satisfaction survey of the Primary Health Care course of the Institute of Health for Well-being]. / Análisis mixto de la encuesta de satisfacción del curso de Atención Primaria de Salud del Instituto de Salud para el Bienestar.

Background: Describing the perception towards the online course on Primary Health Care (PHC) of the Institute of Health for Well-being (INSABI) will allow to establish improvement actions. Objective: Describe the factors that contribute to satisfaction with the PHC course offered online by INSABI. Material and methods: 620 records of the Health Education System for Well-being were studied. Satisfaction was determined using a Likert-type questionnaire with three dimensions: virtual environment, cognitive area, and measurement of learning. A deductive analysis of the open opinions was carried out. Results: 70% of the health personnel approved the course in less than a week, with an initial score of 5.41 ±1.9 points and final score of 7.8 ± 1.2. More than 65% had scores above the average in the three dimensions. Satisfaction with the virtual environment was 15.57 ± 3.4 points, and 15.73 ± 3.3 with the cognitive dimension. Age and gender were associated with dissatisfaction with the virtual environment and in the cognitive dimension, age was associated with dissatisfaction; 27.7% expressed negative comments, 28.5% related to course extension; 15.5% about the didactic techniques, 10.9% about the speakers and 10.4% about the final exam. Conclusions: The course generates significant learning, 62.4% of the students have a positive or neutral opinion. However, 27.8% expressed dissatisfaction, the majority related to the extension of the course.

Introducción: describir la percepción hacia el curso en línea de Atención Primaria de Salud (APS) del Instituto de Salud para el Bienestar (INSABI) permitirá establecer acciones de mejora. Objetivo: conocer el grado de satisfacción con el curso sobre APS que el INSABI ofrece en línea mediante un análisis mixto. Material y métodos: se estudiaron 620 registros del Sistema Educativo de Salud para el Bienestar. La satisfacción se determinó mediante un cuestionario tipo Likert con tres dimensiones: ambiente virtual, área cognitiva y medición del aprendizaje. Se realizó un análisis deductivo de las opiniones abiertas. Resultados: el 70% de los trabajadores de la salud aprobaron el curso en menos de una semana con una calificación inicial de 5.41 ± 1.9 puntos y la final de 7.8 ± 1.2. Más del 65% tuvieron puntuaciones por arriba de la media en las tres dimensiones. La satisfacción con el ambiente virtual fue de 15.57 ± 3.4 puntos, y de 15.73 ± 3.3 con la dimensión cognitiva. La edad y el sexo se asociaron a insatisfacción con el ambiente virtual y en la dimensión cognitiva, la edad se asoció con insatisfacción; el análisis cualitativo mostró que 27.7% expresaron comentarios negativos, 28.5% relacionadas con la extensión del curso; 15.5% sobre las técnicas didácticas, 10.9% acerca de los ponentes y 10.4% sobre el examen final. Conclusiones: el curso genera aprendizaje significativo, el 62.4% de los educandos tienen una opinión positiva o neutra. Sin embargo, 27.8% manifestaron insatisfacción, la mayoría relacionada con la extensión del curso.

[Development of the INSABI educational strategy: a lesson learned from the COVID-19 epidemic]. / Desarrollo de la estrategia educativa del INSABI: una lección aprendida a partir de la epidemia de COVID-19.

The Institute for Health for Well-being (INSABI according to its initials in Spanish), in collaboration with the National Institute of Medical Sciences and Nutrition Salvador Zubirán (INCMNSZ), instituted the Continuous Training on clinical management "Mexico against COVID-19" in 2020, with the purpose of training the frontline health personnel in the care for patients with COVID-19 in the context of hospital reconversion through the COVIDUTI platform. Virtual conferences were held for medical personnel from all over the country with the possibility of interacting with various specialists. In 2020, 215 sessions were held and 158 in 2021. That year educational content was expanded and included topics for other health categories, such as nursing and social work. In October 2021, it was established the Health Educational System for Well-being (SIESABI), with the aim of promoting continuous and permanent education for health workers. It currently offers face-to-face and virtual courses, permanent seminars, and telementoring, with the possibility of providing academic follow-up to its subscribers and linking priority courses that are on other platforms. The educational platform is an opportunity to unify the efforts of the health system in Mexico in the continuous and permanent education of professionals who care for people without social security and thereby contribute to the implementation of a model of care based on primary health care (PHC).

El Instituto de Salud para el Bienestar (INSABI), en colaboración con el Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán (INCMNSZ), instituyó el entrenamiento continuo sobre manejo clínico "México contra COVID-19" en 2020, con el propósito de capacitar al personal de salud de primera línea en la atención de pacientes con COVID-19 en el contexto de la reconversión hospitalaria mediante la plataforma COVIDUTI. Se realizaron conferencias virtuales dirigidas a personal médico de todo el país con la posibilidad de interactuar con diversos especialistas. En 2020 se realizaron 215 sesiones y 158 en 2021. Ese año se ampliaron contenidos educativos y se incluyeron temas para otras categorías de salud, como enfermería y trabajo social. En octubre de 2021 se estableció el Sistema Educativo de Salud para el Bienestar (SIESABI), con el objetivo de promover la educación continua y permanente para los trabajadores de la salud. Actualmente ofrece cursos presenciales, virtuales, seminarios permanentes y telementorías, con la posibilidad de dar seguimiento académico a sus suscriptores y vincular cursos prioritarios que están en otras plataformas. La plataforma educativa es una oportunidad para unificar los esfuerzos del sistema de salud en México en la educación continua y permanente de los profesionales que atienden a personas sin seguridad social y, con ello, contribuir en la implementación de un modelo de atención basado en Atención Primaria de Salud (APS).

Genomic insights of Leclercia adecarboxylata strains linked to an outbreak in public hospitals in Mexico.

BACKGROUND: Leclercia adecarboxylata is a bacteria closely related to Escherichia coli according to its biochemical characteristics and is commonly considered non-pathogenic although a growing number of publications classify it as an emerging pathogen. Fosfomycin resistance is a common trait for L. adecarboxylata encoded by fosALA gene. OBJECTIVE: To analyze genomic traits of sixteen L. adecarboxylata strains isolated from blood culture and a bottle of total parenteral nutrition. METHODS: Twenty-eight L. adecarboxylata strains isolated from blood culture and a bottle of total parenteral nutrition were identified biochemically with a Vitek ® automated system. The strains were phenotyped by their growth on Eosin Methylene Blue agar or MacConkey agar plates. Additionally, Pulsed field gel electrophoresis (PFGE) was performed to establish the clonal relationship. The genomic DNA of sixteen strains was obtained using a Qubit ® dsDNA HS Assay Kit and sequenced on an Illumina ® MiSeq instrument. Draft genomes were assembled using PROKKA and Rast. Assemblies were submitted to Resfinder and PathogenFinder from the Center for Genomic Epidemiology in order to find resistance genes and pathogenic potential. IslandViewer4 was also used to find Pathogenicity and Phage Islands. For identification of the fosA gene, manual curation and Clustal analysis was performed. A novel FosA variant was identified. Finally, phylogenetic analysis was performed using VAMPhyRE software and Mega X. RESULTS: In this paper, we report the genomes of sixteen strains of Leclercia adecarboxylata causing an outbreak associated with parenteral nutrition in public hospitals in Mexico. The genomes were analyzed for genetic determinants of virulence and resistance. A high pathogenic potential (pathogenicity index 0.82) as well as multiple resistance genes including carbapenemics, colistin and efflux pumps were determined. Based on sequence analysis, a new variant of the fosALA gene was described. Finally, the outbreak was confirmed by establishing the clonal relationship among the sixteen genomes obtained. CONCLUSIONS: Commensal strains of L. adecarboxylata may acquire genetic determinants that provide mechanisms of host damage and go unnoticed in clinical diagnosis. L. adecarboxylata can evolve in a variety of ways including the acquisition of resistance and virulence genes representing a therapeutic challenge in patient care.

Pandemic influenza A/H1N1 virus infection and TNF, LTA, IL1B, IL6, IL8, and CCL polymorphisms in Mexican population: a case-control study.

BACKGROUND: Some patients have a greater response to viral infection than do others having a similar level of viral replication. Hypercytokinemia is the principal immunopathological mechanism that contributes to a severer clinical course in cases of influenza A/H1N1. The benefit produced, or damage caused, by these cytokines in severe disease is not known. The genes that code for these molecules are polymorphic and certain alleles have been associated with susceptibility to various diseases. The objective of the present study was to determine whether there was an association between polymorphisms of TNF, LTA, IL1B, IL6, IL8, and CCL1 and the infection and severity of the illness caused by the pandemic A/H1N1 in Mexico in 2009. METHODS: Case-control study. The cases were patients confirmed with real time PCR with infection by the A/H1N1 pandemic virus. The controls were patients with infection like to influenza and non-familial healthy contacts of the patients with influenza. Medical history and outcome of the disease was registered. The DNA samples were genotyped for polymorphisms TNF rs361525, rs1800629, and rs1800750; LTA rs909253; IL1B rs16944; IL6 rs1818879; IL8 rs4073; and CCL1 rs2282691. Odds ratio (OR) and the 95% confidence interval (95% CI) were calculated. The logistic regression model was adjusted by age and severity of the illness in cases. RESULTS: Infection with the pandemic A/H1N1 virus was associated with the following genotypes: TNF rs361525 AA, OR = 27.00; 95% CI = 3.07-1248.77); LTA rs909253 AG (OR = 4.33, 95% CI = 1.82-10.32); TNF rs1800750 AA (OR = 4.33, 95% CI = 1.48-12.64); additionally, LTA rs909253 AG showed a limited statistically significant association with mortality (p = 0.06, OR = 3.13). Carriers of the TNF rs1800629 GA genotype were associated with high levels of blood urea nitrogen (p = 0.05); those of the TNF rs1800750 AA genotype, with high levels of creatine phosphokinase (p=0.05). The IL1B rs16944 AA genotype was associated with an elevated number of leukocytes (p <0.001) and the IL8 rs4073 AA genotype, with a higher value for PaO2 mm Hg. CONCLUSION: The polymorphisms of genes involved in the inflammatory process contributed to the severity of the clinical behavior of infection by the pandemic influenza A/H1N1 virus.

Functional state analysis of phagocytic cells of patients with type 2 diabetes and pulmonary tuberculosis.

BACKGROUND: The phagocytic function in pulmonary tuberculosis (PTB) and Type 2 diabetes (T2D) has been explored mainly in macrophages but not in polymorphonuclears (PMN). The purpose of this study was to determine the functional status of PMN leukocytes in patients with pulmonary tuberculosis (PTB), Type 2 diabetes (T2D), and in patients with both diseases. METHODS: An observational, prospective, and comparative study was carried out. 30 ambulatory patients with T2D, 10 with PTB undergoing treatment and 10 patients with PTB and T2D, and 44 healthy subjects were studied. PMN leukocytes were separated, the capacity of these cells to produce hydrogen peroxide and to reduce nitroblue tetrazolium (NBT) in response to stimulus with the phorbolic ester of myristic acid (PMA) was measured; and the capacity of PMN leukocytes to adhere to surfaces was determined. RESULTS: Concerning the test for adherence, on comparing healthy subjects with patients with T2D+PTB, we observed a clear decrease in cellular adherence in the group of patients with both diseases; it was statistically significant (p = 0.007).With regard to phagocytic function, we observed that in NBT reduction as well as in hydrogen peroxide production, statistically significant differences were not obtained on comparing healthy subjects with any of the three groups of patients. CONCLUSIONS: We observed a clear decrease in cellular adherence when both diseases co-exist. These results could indicate the need for the co-existence of T2D and TB to cause deterioration in the cells' adherence activity. The microtechniques employed permit the evaluation in a practical manner of certain phagocytic-activity expressions.

[CSF RT-PCR standardization in pediatric patients with enteroviral central nervous system infection]. / Estandarización de RT-PCR en LCR de pacientes pediátricos con infección del sistema nervioso central por enterovirus.

OBJECTIVE: We standardized the RT-PCR panviral CSF and determined its applicability in detecting acute enterovirus infection in the central nervous system in children under 15 years. MATERIAL AND METHODS: RT-PCR was performed directly in CSF samples of 10 pediatric patients with suspected CNS infection and 9, with different conditions of the central nervous system. RESULTS: 80% (8/10) of RT-PCR samples were positive for enterovirus in patients with suspected CNS infection and no sample was positive in patients with different ailments. CONCLUSIONS: Since enteroviruses are among the main etiologies of pediatric encephalitis, RT-PCR could be particularly useful for rapid detection in CSF.

The usefulness of the buffy coat smear and panbacterial polymerase chain reaction in early diagnosis of neonatal sepsis.

OBJECTIVE: In this study are evaluated the usefulness of the buffy coat smear and panbacterial polymerase chain reaction (PCR) as diagnostic tests in the early detection of neonatal sepsis. MATERIAL AND METHODS: It was studied 49 patients aged up to 28 days who were hospitalized in the Intensive Care Unit (ICUs) of the Neonatology, with a clinical diagnosis of neonatal sepsis and 49 umbilical cord samples of healthy newborns. Blood cultures and 50 microL of plasma were taken for the DNA and performance of the broad-range PCR primer system (panbacterial PCR). Simultaneously, were taken three capillaries with blood for the leukocyte layer (buffy coat) smear, we performed three stains: Gram; Löeffler blue methylene (LBM), and acridine orange (AO). Statistical analysis included sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) against the clinical diagnosis. RESULTS: With respect to stains of buffy coat smear, they resulted very specific, from 90-97%, with 64-75% sensitivity, 87-94% PPV, and 77-82% NPV. In inverse fashion, PCR resulted very sensitive at 96%, with 91% specificity, 92% PPV, and 96% NPV. CONCLUSIONS: Buffy coat smear stains are easy, fast, and specific, while that of PCR was highly sensitive. Thus, both can be utilized as diagnostic tests.

[Pro-inflammatory cytokines related to severity and mortality in type 2 diabetes patients with soft tissue infection]. / Citocinas, diabetes e infección de tejidos blandos. Su relación con la severidad y mortalidad.

OBJECTIVE: to determine the relation between IL6, IL10 and TNFa serum levels in a cohort of patients with type 2 diabetes (T2D) and severe soft tissue infections (STI), with severity and mortality factors. METHODS: A. comparative and transversal, study with 15 adult patients, any gender, with T2D and STI were done. A T2D control group of 20 patients without STI was included. Apache II Score, glycemia and by ELISA, IL6, IL10 and TNFa, were determined. RESULTS: in all patients, it was a correlation at beginning between glycemia and IL6 (r = 0.67, IC 95 % 0.24-0.88), as soon as glycemia and Apache II, (r = 0.59, IC 95 % 0.11-0.83). CONCLUSIONS: although IL6 was very usefulness, it is not a routine test in clinical laboratory and it is expensive, but in medical practice, it could be possible to evaluate these patients with Apache II Score and glycemia. However, in STI, the values of IL6 and IL10 were highly significant. It is likely that IL6 is a marker of poor outcome.