Stroke etiologies in patients with COVID-19: the SVIN COVID-19 multinational registry.

BACKGROUND AND PURPOSE: Coronavirus disease 2019 (COVID-19) is associated with a small but clinically significant risk of stroke, the cause of which is frequently cryptogenic. In a large multinational cohort of consecutive COVID-19 patients with stroke, we evaluated clinical predictors of cryptogenic stroke, short-term functional outcomes and in-hospital mortality among patients according to stroke etiology. METHODS: We explored clinical characteristics and short-term outcomes of consecutively evaluated patients 18 years of age or older with acute ischemic stroke (AIS) and laboratory-confirmed COVID-19 from 31 hospitals in 4 countries (3/1/20-6/16/20). RESULTS: Of the 14.483 laboratory-confirmed patients with COVID-19, 156 (1.1%) were diagnosed with AIS. Sixty-one (39.4%) were female, 84 (67.2%) white, and 88 (61.5%) were between 60 and 79 years of age. The most frequently reported etiology of AIS was cryptogenic (55/129, 42.6%), which was associated with significantly higher white blood cell count, c-reactive protein, and D-dimer levels than non-cryptogenic AIS patients (p

COVID-19 prevalence and mortality in people with epilepsy: A nation-wide multicenter study.

BACKGROUND: To assess the prevalence, severity, and mortality of COVID-19 in people with epilepsy (PWE) and evaluate seizure control in PWE during and after COVID-19. METHODS: Retrospective, observational, multicenter study conducted in 14 hospitals. Medical records of randomly selected PWE followed at neurology outpatient clinics were reviewed. Proportion of PWE with a positive test for SARS-CoV-2 during 2020 was calculated. Risk factors associated with COVID-19 and its morbimortality were evaluated. RESULTS: 2751 PWE were included, mean age 48.8 years (18-99), 72.4% had focal epilepsy, and 35% were drug-refractory. COVID-19 prevalence in PWE was 5.53%, while in the Spanish population was 4.26%. Proportion of admissions to hospital, ICU, and deaths in PWE were 17.1%, 2%, and 4.61% of COVID-19 cases, while in Spanish population were 10.81%, 0.95%, and 2.57%, respectively. A severe form of COVID-19 occurred in 11.8%; dyslipidemia, institutionalization at long-term care facilities, intellectual disability, and older age were associated risk factors. Older age, hypertension, dyslipidemia, cardiac disease, and institutionalization were associated with mortality from COVID-19. Seizure control was stable in 90.1% of PWE during acute COVID-19, while 8.6% reported an increase in seizure frequency. During post-COVID-19 follow-up, 4.6% reported seizure control worsening. CONCLUSIONS: COVID-19 was moderately prevalent in PWE. One out of 5 patients required medical attention and 4.6% died due to COVID-19. Older age, dyslipidemia, institutionalization, and intellectual disability were significant risk factors associated with severe COVID-19. Seizure control remained stable during COVID-19 and throughout long-term follow-up in most PWE who contracted the infection.

Resurgence of Vaccine-Preventable Diseases in Venezuela as a Regional Public Health Threat in the Americas.

Venezuela's tumbling economy and authoritarian rule have precipitated an unprecedented humanitarian crisis. Hyperinflation rates now exceed 45,000%, and Venezuela's health system is in free fall. The country is experiencing a massive exodus of biomedical scientists and qualified healthcare professionals. Reemergence of arthropod-borne and vaccine-preventable diseases has sparked serious epidemics that also affect neighboring countries. In this article, we discuss the ongoing epidemics of measles and diphtheria in Venezuela and their disproportionate impact on indigenous populations. We also discuss the potential for reemergence of poliomyelitis and conclude that action to halt the spread of vaccine-preventable diseases within Venezuela is a matter of urgency for the country and the region. We further provide specific recommendations for addressing this crisis.

Single-Nucleotide Polymorphism 309T>G in the MDM2 Promoter Determines Functional Outcome After Stroke.

Background and Purpose- The E3 ubiquitin ligase MDM2 (murine double minute 2) is the main negative regulator of the p53 protein-a key player in neuronal apoptosis after ischemia. A functional single-nucleotide polymorphism in the human MDM2 gene promoter (rs2279744) regulates MDM2 protein expression. We investigated whether the MDM2 SNP309, by controlling p53-mediated apoptosis, determines functional outcome after stroke. Methods- Primary cortical neurons were subjected to oxygen and glucose deprivation. Mice were subjected to ischemic (transient middle cerebral artery occlusion) or hemorrhagic (collagenase injection) stroke models. Protein and mRNA levels of MDM2 and p53 were measured in both neuronal and brain extracts. The interaction of MDM2 with p53 was disrupted by neuronal treatment with nutlin-3a. siRNA was used to knockdown MDM2 expression. We analyzed the link between the MDM2 SNP309 and functional outcome, measured by the modified Rankin Scale scores, in 2 independent hospital-based stroke cohorts: ischemic stroke cohort (408 patients) and intracerebral hemorrhage cohort (128 patients). Results- Experimental stroke and oxygen and glucose deprivation induced the expression of MDM2 in the brain and neurons, respectively. Moreover, oxygen and glucose deprivation promoted MDM2 binding with p53 in neurons. Disruption of the MDM2-p53 interaction with nutlin-3a, or MDM2 knockdown by siRNA, triggered p53 accumulation, which increased neuronal susceptibility to oxygen and glucose deprivation-induced apoptosis. Finally, we showed that patients harboring the G allele in the MDM2 promoter had higher MDM2 protein levels and showed better functional outcome after stroke than those harboring the T/T genotype. The T/T genotype was also associated with large infarct volume in ischemic stroke and increased lesion volume in patients with intracerebral hemorrhage. Conclusions- Our results reveal a novel role for the MDM2-p53 interaction in neuronal apoptosis after ischemia and show that the MDM2 SNP309 determines the functional outcome of patients after stroke.

Double or nothing: red flag symptoms of critical carotid stenosis, a case report.

BACKGROUND: Detailed knowledge of every possible manifestation of Internal Carotid Artery (ICA) disease is important. For improving detection and a timely adoption of secondary prevention procedures or treatments. Transient oculomotor nerve palsies have been described associated with stenosis or occlusion of the ICA. CASE PRESENTATION: We described a patient that develop a sequential combination of transient monocular loss of vision followed by binocular diplopia secondary to an unstable atherosclerotic preocclusive stenosis of an internal carotid artery previously treated with radiotherapy. CONCLUSIONS: The peculiar sequence of transient monocular vision that give rise later into a transient binocular diplopia (double or nothing) should be kept in mind as a possible manifestation of critical stenosis of ICA.

[Prevalence of beta-lactamase CTX-M-15 in phylogenetic groups of uropathogenic Escherichia coli isolated from patients in the community of Merida, Venezuela]. / Prevalencia de beta-lactamasa CTX-M-15 en grupos filogenéticos de Escherichia coli uropatógena aisladas en pacientes de la comunidad en Mérida, Venezuela.

In this study we determined the prevalence of extended-spectrum beta-lactamases (ESBLs) in phylogenetic groups of uropathogenic E. coli (UPEC) isolated from patients in the community. Twenty one UPEC strains with reduced susceptibility to broad-spectrum cephalosporins were collected between January 2009 and July 2010, from patients with urinary tract infection who attended the Public Health Laboratory in Mérida, Venezuela. Genotypic characterization determined that all UPEC strains harbored blaBLEEs genes: 76.2% of the strains showed the presence of a single ESBL-producer gene, represented by blaCTX-M-15, whereas 23.8% of UPEC showed various combinations of bla genes (blacCTX-M-15 + blaTEM-1, blaCTX-M-15 + blaSHV and blaSHV + blaTEM-1). In this study, 61.9% of the isolates were placed in phylogroup A and the remaining strains were assigned to group B2 (38.1%). There was no evidence of spread of a particular UPEC clone; only seven strains belonged to a clonal group with an index of similarity greater than 85%. To our knowledge, this is the first description of blxCTX-M-15 in UPEC from patients with community-acquired urinary tract infections, which shows that Venezuela is also part of the so-called CTX-M-15 pandemic. The findings in this study, as well as its clinical and epidemiological implications, lead to the need for monitoring and controlling the spread of CTX-M-15 producing UPECs, not only regionally, but also nationwide.

[Distribution of phylogenetic groups and virulence factors in CTX-M-15 ß-lactamase-producing uropathogenic Escherichia coli strains isolated from patients in the community of Mérida, Venezuela]. / Distribución de grupos filogenéticos y factores de virulencia en cepas de Escherichia coli uropatógena productora de ß-lactamasa CTX-M-15 aisladas de pacientes de la comunidad en Mérida, Venezuela.

In this study, the distribution of phylogenetic groups and the genetic detection of virulence factors in CTX-M-15 ß-lactamase-producing uropathogenic Escherichia coli (UPEC) strains were analyzed. Twenty eight strains were isolated between January 2009 and July 2011 from patients with urinary tract infection (UTI) who attended the Public Health Laboratory at Mérida, Venezuela. Determination of phylogenetic groups and detection of six virulence genes, fimH, fyuA, kpsMTII, usp, PAI and papAH, were performed by PCR amplification. Fifteen of the 28 isolates were mainly located in the phylogenetic group A, followed by B2 (12/28) and D (1/28). No direct relationship between the severity or recurrence of UTI and the distribution of phylogroups was observed. All studied virulence factors were found in group B2 strains with the highest frequency. The prevalent virulence profile included the combination of three main genes: fimH, kpsMTII and fyuA and, to a lesser extent, the presence of other determinants such as usp, PAI and/or papAH. These results indicate that virulent UPEC incorporated three important properties: adhesion, iron uptake and evasion of phagocytosis, which favored the production of recurrent UTI. This is the first report describing the association of phylogenetic groups with the potential virulence of CTX-M-15 ß-lactamase producing UPEC strains in Venezuela.

Genetic diversity of Tn916-related transposons among drug-resistant Streptococcus pneumoniae isolates colonizing healthy children in Venezuela.

Among a collection of 48 multidrug-resistant pneumococcal strains colonizing healthy children in a small municipality of Mérida, Venezuela, we identified sequence types (STs) related to a variety of internationally spreading drug-resistant clones, as well as ST135, thus far isolated only in Europe. The clones invariably harbored one or more of the Tn916-related transposons Tn3872, Tn5253, Tn6002, Tn2009, and Tn2010. Finally, our data suggest both structural rearrangements in certain transposons and occurrence of novel transposable elements.

CTX-M-14 ß-lactamase-producing Citrobacter freundii isolated in Venezuela.

A clinical isolate of C. freundii with reduced susceptibility to extended-spectrum ß-lactams from a woman with cystocele associated with recurrent urinary tract infection was analyzed. Susceptibility tests, double disk synergy tests (DDST) and enzymatic activity by the agar iodometric method suggested the presence of ESBLs. Conjugation experiments revealed the presence of a large conjugative plasmid (pLM07/20) with an exclusive FrepB replicon type (IncF/FIB). PCR analysis and sequencing confirmed the presence of the blaCTX-M-14 gene in the pLM07/20 from C. freundii.LM07/10. Although this is the first report of CTX-M-14 in Venezuela, we alert the medical community that future increase of these ß-lactamases in our city could be due to dissemination of plasmids into bacterial populations.

Fibrin-Platelet Clots in Acute Ischemic Stroke. Predictors and Clinical Significance in a Mechanical Thrombectomy Series.

Introduction: The histological composition of the clot influences its mechanical properties, affects the efficacy of endovascular treatment (EVT), and could determine the clinical outcome of patients with acute ischemic stroke (AIS). Insights into clot composition may guide therapeutic decision-making prior to EVT and facilitate revascularization therapies. Material and Methods: Consecutive patients with AIS recorded in a prospective single-center reperfusion registry from December 2015 to December 2019 and treated with EVT were included. Baseline, laboratory [including post-procedural C-reactive protein (CRP)], radiological, and angiographic variables were analyzed. We aimed to study the relationship between histological composition of the clot with basal neuroimaging, laboratory markers, and recanalization technique. The secondary outcome was to analyze the correlation between clot composition and functional outcome at 3 months assessed by the modified Rankin scale (mRS). Results: From the study period, 360 AIS patients treated with EVT were included, of whom 189 (53%) fulfilled the inclusion criteria. One hundred (53%) cases of fibrin-predominant clot (FPC) were recorded. Full recanalization in FPC cases was achieved with higher probability when stent retrievers (SR) were selected as the first-line device (68.2%, p = 0.039). Patients with FPC had higher levels of CRP (p = 0.02), lower frequency of the hyperdense middle cerebral artery (HMCA) in baseline imaging (p = 0.039), and higher rates of mortality (p = 0.012). The multivariate analysis showed that the absence of HMCA (OR = 0.420; 95% CI 0.197-0.898; p = 0.025) and higher levels of CRP (OR = 1.01; 95% CI 1.003-1.019; p = 0.008) were predictors of FPC. Leukocytes and platelet counts were not associated with clot histology. Conclusions: The absence of HMCA and higher levels of CRP were markers of FPC. In patients with FPC, complete recanalization was most likely to be achieved when a SR was selected as first line of treatment. Mortality was higher in patients within this histologic group.

[Distribution of phylogenetic groups, virulence factors and antimicrobial susceptibility in strains of uropathogenic Escherichia coli]. / Distribución de grupos filogenéticos, factores de virulencia y susceptibilidad antimicrobiana en cepas de Escherichia coli uropatógena.

BACKGROUND: The difference between the pathogenic isolates and commensals of Escherichia coli is based on their phylogenetic antecedents. In Venezuela there are few studies that describe the pathogenic potential of phylogenetic groups in E. coli. AIMS: Relate antimicrobial susceptibility, distribution of phylogenetic groups and virulence genes in strains of uropathogenic E. coli (ECUP) isolated from patients with UTI. METHODS: We studied 17 ECUP strains, isolated from adult patients hospitalized in two health institutions. The susceptibility to 8 antibiotics was determined by the broth microdilution (MDC) method. Extended spectrum ß-lactamases (ESBL) and carbapenemases were phenotypically detected. The phylogenetic groups and the detection of the virulence genes were determined by PCR. RESULTS: All strains synthesized ESBL and of these, 41% were associated with the production of a carbapenemases (KPC or MBL). The phylogroup B2 (41%) was the most predominant. The most frequent virulence genes were fimH and fyuA with 82% each. Only one strain from group F was positive to the 6 genes studied. DISCUSSION: The diversity of associations between virulence genes and resistance profiles in the ECUP are evolving continuously, their distribution in the different phylogenetic groups depends to a large extent on the clinical epidemiological characteristics of the study groups.

Oral Anticoagulation and Risk of Symptomatic Hemorrhagic Transformation in Stroke Patients Treated With Mechanical Thrombectomy: Data From the Nordictus Registry.

Introduction: We aimed to evaluate if prior oral anticoagulation (OAC) and its type determines a greater risk of symptomatic hemorrhagic transformation in patients with acute ischemic stroke (AIS) subjected to mechanical thrombectomy. Materials and Methods: Consecutive patients with AIS included in the prospective reperfusion registry NORDICTUS, a network of tertiary stroke centers in Northern Spain, from January 2017 to December 2019 were included. Prior use of oral anticoagulants, baseline variables, and international normalized ratio (INR) on admission were recorded. Symptomatic intracranial hemorrhage (sICH) was the primary outcome measure. Secondary outcome was the relation between INR and sICH, and we evaluated mortality and functional outcome at 3 months by modified Rankin scale. We compared patients with and without previous OAC and also considered the type of oral anticoagulants. Results: About 1.455 AIS patients were included, of whom 274 (19%) were on OAC, 193 (70%) on vitamin K antagonists (VKA), and 81 (30%) on direct oral anticoagulants (DOACs). Anticoagulated patients were older and had more comorbidities. Eighty-one (5.6%) developed sICH, which was more frequent in the VKA group, but not in DOAC group. OAC with VKA emerged as a predictor of sICH in a multivariate regression model (OR, 1.89 [95% CI, 1.01-3.51], p = 0.04) and was not related to INR level on admission. Prior VKA use was not associated with worse outcome in the multivariate regression model nor with mortality at 3 months. Conclusions: OAC with VKA, but not with DOACs, was an independent predictor of sICH after mechanical thrombectomy. This excess risk was associated neither with INR value by the time thrombectomy was performed, nor with a worse functional outcome or mortality at 3 months.

[Phenotypic variations of susceptibility in Klebsiella pneumoniae strains of nosocomial origin and their association with biofilm formation]. / Variaciones fenotipicas de susceptibilidad en cepas de Klebsiella pneumoniae de origen nosocomial y su asociación con la formación de biopelículas.

Klebsiella pneumoniae is the principal opportunist pathogen associated with nosocomial infections. The morbidity and mortality produced by this microorganism is related mostly to antimicrobial resistance and its capacity to form biofilms. In this study, the phenotypic variations of susceptibility in 50 Klebsiella pneumoniae strains (26 betaLEE-producer and 24 no betaLEE-producer) isolated from pediatric patients with nosocomial septicemia at the High-risk Neonatal Unit of the Instituto Aut6nomo Hospital Universitario de Los Andes, Mérida, Venezuela and their associations with biofilm formation were analyzed. These strains were biochemically identified by standard microbiology methods. Antimicrobial susceptibility was determined by minimal inhibitory concentration and biofilm formation was detected by spectrophotometric techniques. Most of betaLEE strains were frequently associated with resistance markers such as tobramycin, kanamycin, tetracycline, chloramphenicol (38% each), streptomycin (36%) and netilmicin (34%). The predominant multiresistent phenotype was composed of betaLEE+ -AMG-TET-CLF (30%). 18% of K. pneumoniae strains were sensible to the antibiotics tested. Formation of biofilm was observed in 77.8% and 96.2% of sensible and resistent strains, respectively. There was no statistic relation between phenotipic variations of susceptibility and biofilm formation (p > = 0.05). However, both characteristics of K. pneumoniae should be evaluated to determine the therapeutic conduct to treat patients with nosocomial infection.

[blaVIM-2 gene detection in metallo-beta-lactamase-producing Pseudomonas aeruginosa strains isolated in an intensive care unit in Ciudad Bolívar, Venezuela]. / Detección del gen blaVIM-2 en cepas de Pseudomonas aeruginosa productoras de metalo beta-lactamasa aisladas en una unidad de cuidados intensivos en Ciudad Bolívar, Venezuela.

Ten Pseudomonas aeruginosa strains with resistance to broad-spectrum cephalosporin and carbapenems were studied to determine the presence of genes that mediate the production of metallo-beta-lactamases. These strains were isolated from patients with nosocomial infection at the Intensive Care Unit of the Complejo Hospitalario "Ruiz y Paéz" of Ciudad Bolívar, Bolívar State, Venezuela, from 2003 to 2006. In all isolates a metallo-enzyme activity was detected by using the double disk synergism test. PCR amplification of genes encoding the families IMP, VIM and SPM metallo-beta-lactamases showed the presence of a blaVIM gene in all strains studied. DNA sequencing revealed that all isolates showed the presence of blaVIM-2. These results suggest that it is necessary to keep these strains under epidemiologic surveillance, establish laboratory strategies for opportune detection and the implementation of new policies to ensure the appropriate use of antibiotics in this institution.

Molecular typing, antibiotic resistance profiles and biocide susceptibility in Salmonella enterica serotypes isolated from raw chicken meat marketed in Venezuela.

INTRODUCTION: Salmonella is a common bacterial cause of foodborne diarrhea worldwide. The purpose of this study was to characterize antimicrobial resistance and susceptibility to biocides in Salmonella enterica serotypes isolated from raw chicken meat, as well as to study the genetic relationship between strains and virulence profiles. METHODS: Nine Salmonella enterica strains (5 S. Heidelberg; 2 S. Enteritidis; 1 S. Typhimurium and 1 S. Meleagridis) recovered from raw chicken meat marketed in the urban area of Mérida, Venezuela, were studied. Phenotypic characterization was based on antimicrobial susceptibility testing as well as detection of extended-spectrum ß-lactamases (ESBLs) by double-disc synergy. The susceptibility to biocides was determined using the dilution-neutralization methods. The detection of quinolone resistance-determining regions of gyrA, gyrB, and parC genes, bla ESBLs genes, plasmid-mediated quinolone resistance determinants and virulence genes (invA and spvC) was carried out by PCR. All strains were typed using PFGE. RESULTS: Multidrug-resistance was evident in 6 of 9 strains studied. However, all Salmonella serotypes were susceptible to the tested biocides. Genotypic characterization determined that 5 strains harbored the bla CTXM-2, 4 bla TEM-1 and 3 qnrB19 genes. All strains were positive for the invA gene. The spvC gene was detected in 4 of them. PFGE grouped Salmonella strains into 4 different patterns that represented individual serotypes. CONCLUSIONS: This study provides valuable information on antibiotic resistance, biocide susceptibility profiles, virulence gene content and genetic diversity of Salmonella enterica serotypes isolated from raw chicken meat marketed in Venezuela, and evidenced a health risk for consumers.

Seroprevalence and molecular detection of Toxoplasma gondii among donors in a blood bank in Cúcuta, Colombia Introduction: Toxoplasma gondii infection / Seroprevalencia y detección molecular de Toxoplasma gondii en donantes de un banco de sangre de Cúcuta, Colombia

Introduction: Toxoplasma gondii infection manifests differently in humans according to their immunity ranging from asymptomatic profiles to severe disease. There are multiple transmission mechanisms including blood transfusions, but little is known about the frequency of T. gondii infection in Colombia's blood banks. Objective: To determine the prevalence of T. gondii infection in blood donors of a blood bank in the city of Cúcuta by serological and molecular diagnostic techniques. Materials and methods: We identified IgG and IgM antibodies against T. gondii by immunoassay in serum from 348 donors. The frequency of T. gondii DNA was determined by polymerase chain reaction (PCR) in whole blood from seropositive donors and relevant variables were analyzed based on the information obtained from surveys during blood donor selection. Results: Out of the 348 enrolled donors, 134 (38.5%) showed IgG antibodies against T. gondii; two of them (0.6%) had both IgG and IgM, and in two of them (1.5%), parasite DNA was detected in blood samples. A bivariate analysis indicated an association between seropositivity to T. gondii and being over 26 years of age (p=0.020). Conclusions: The prevalence of T. gondii infection found in the blood donors of this study suggests a significant exposure to the infectious agent that becomes relevant when parasitemia is detected.

Introducción. La infección por Toxoplasma gondii puede presentarse en los humanos con un amplio rango de manifestaciones que van desde el estado asintomático hasta la enfermedad grave, según el estado inmunológico del individuo. Los mecanismos de transmisión incluyen la transfusión sanguínea, pero poco se sabe sobre la frecuencia del parásito en los bancos de sangre de Colombia. Objetivo. Determinar la prevalencia de la infección con T. gondii en donantes de un banco de sangre de Cúcuta mediante técnicas de diagnóstico serológico y molecular. Materiales y métodos. Se determinaron los anticuerpos IgG e IgM contra T. gondii mediante un inmunoensayo en suero en 348 donantes. Se determinó la frecuencia de ADN de T. gondii utilizando la reacción en cadena de la polimerasa (PCR) en sangre total de donantes seropositivos y se analizaron las variables de interés con base en la información obtenida durante la selección de donantes. Resultados. De los 348 donantes participantes, 134 (38,5 %) presentaron anticuerpos IgG contra T. gondii; dos (0,6 %) de ellos presentaron tanto IgG como IgM y, en dos (1,5 %), se detectó ADN del parásito en la sangre. Un análisis bivariado evidenció una asociación entre la seropositividad para T. gondii y tener más de 26 años de edad (p=0,020). Conclusiones. La prevalencia de la infección con T. gondii encontrada en los donantes de sangre sugiere una exposición significativa al agente, la cual adquiere relevancia al detectarse la parasitemia.

The Neuronal Ischemic Tolerance Is Conditioned by the Tp53 Arg72Pro Polymorphism.

Cerebral preconditioning (PC) confers endogenous brain protection after stroke. Ischemic stroke patients with a prior transient ischemic attack (TIA) may potentially be in a preconditioned state. Although PC has been associated with the activation of pro-survival signals, the mechanism by which preconditioning confers neuroprotection is not yet fully clarified. Recently, we have described that PC-mediated neuroprotection against ischemic insult is promoted by p53 destabilization, which is mediated by its main regulator MDM2. Moreover, we have previously described that the human Tp53 Arg72Pro single nucleotide polymorphism (SNP) controls susceptibility to ischemia-induced neuronal apoptosis and governs the functional outcome of patients after stroke. Here, we studied the contribution of the human Tp53 Arg72Pro SNP on PC-induced neuroprotection after ischemia. Our results showed that cortical neurons expressing the Pro72-p53 variant exhibited higher PC-mediated neuroprotection as compared with Arg72-p53 neurons. PC prevented ischemia-induced nuclear and cytosolic p53 stabilization in Pro72-p53 neurons. However, PC failed to prevent mitochondrial p53 stabilization, which occurs in Arg72-p53 neurons after ischemia. Furthermore, PC promoted neuroprotection against ischemia by controlling the p53/active caspase-3 pathway in Pro72-p53, but not in Arg72-p53 neurons. Finally, we found that good prognosis associated to TIA within 1 month prior to ischemic stroke was restricted to patients harboring the Pro72 allele. Our findings demonstrate that the Tp53 Arg72Pro SNP controls PC-promoted neuroprotection against a subsequent ischemic insult by modulating mitochondrial p53 stabilization and then modulates TIA-induced ischemic tolerance.

Prevalence of Fecal Carriage of CTX-M-15 Beta-Lactamase-Producing Escherichia coli in Healthy Children from a Rural Andean Village in Venezuela.

OBJECTIVES: Antimicrobial resistant extended-spectrum-ß-lactamase-producing Enterobacteriaceae (ESBL-PE) have been shown to be present in healthy communities. This study examined healthy children from the rural Andean village of Llano del Hato, Mérida, Venezuela, who have had little or no antibiotic exposure to determine the prevalence of fecal carriage of ESBL-producing Escherichia coli (ESBL-EC). METHODS: A total of 78 fecal samples were collected in healthy children aged from 1 to 5 years. ESBL-EC were selected in MacConkey agar plates with cefotaxime and further confirmed by the VITEK 2 system. ESBL were phenotypically detected and presence of bla genes and their variants were confirmed by molecular assays. Determination of phylogenetic groups was performed by PCR amplification. Risk factors associated with fecal carriage of ESBL-EC-positive isolates were analyzed using standard statistical methods. RESULTS: Of the 78 children studied, 27 (34.6%) carried ESBL-EC. All strains harbored the blaCTX-M-15 allele. Of these, 8 were co-producers of blaTEM-1, blaTEM-5, blaSHV-5 or blaSHV-12. Co-resistance to aminoglycosides and/or fluoroquinolones was observed in 9 strains. 51.9% of ESBL-EC isolates were classified within phylogroup A. A significant, positive correlation was found between age (≥2.5 - ≤5 years), food consumption patterns and ESBL-EC fecal carriage. CONCLUSION: This is the first study describing the high prevalence of fecal carriage of ESBL-EC expressing CTX-M-15- among very young, healthy children from a rural Andean village in Venezuela with scarce antibiotic exposure, underlining the importance of this population as a reservoir.

Molecular characterisation of multidrug-resistant pneumococcal clones colonising healthy children in Mérida, Venezuela.

Genetic mechanisms of resistance, clonal composition and the occurrence of pili were analysed in 48 multidrug-resistant (MDR) pneumococci isolated from healthy children in Mérida, Venezuela. Intermediate resistance to penicillin was related to variations in pbp2b and pbp2x. High-level resistance to penicillin as well as low susceptibility to cephalosporins and carbapenems were associated with alterations in pbp1a, pbp2b and pbp2x. Non-ß-lactam resistance was associated with Tn3872, Tn5253, Tn6002 and Tn2010 transposons. Macrolide-resistant strains carried ermB or mefE, but not mefA. Tetracycline- and chloramphenicol-resistant pneumococci carried tetM and cat, respectively. MDR pneumococci were related to six clonal complexes (CCs), largely CC156 or CC15. Limited diversity in pbp2a,2b,2x-RFLP profiles within each clone was observed. Conversely, detection of non-ß-lactam resistance and transposons revealed clear genetic diversity within clones. A group of non-typeable/cpsA-negative pneumococci related to the null capsule clade 1 (NCC1) carrying a Tn2009 element was found. Each NCC1-related strain showed a novel MLST allelic combination and a different pbp2a,2b,2x-RFLP profile. PI-1 (pilus type 1 rrgC gene) was present in most of the MDR pneumococci and its occurrence was commonly homogeneous within each clone. PI-1 was present in all CC242 and CC320 pneumococci, whereas it was absent in all CC37, CC81 and NCC1 isolates. CC156 and CC15 isolates showed variations in the occurrence of PI-1. Both PI-1 and the islet for pilus type 2 were present in CC320 isolates. We provide useful data to follow the evolution of clonal composition, mechanisms of antibiotic resistance and the occurrence of pili among pneumococci circulating in Mérida.

[Genetic diversity of extraintestinal Escherichia coli strains producers of beta-lactamases TEM, SHV and CTX-M associated with healthcare].

INTRODUCTION: There are few reports from Venezuela describing the genetic basis that sustains the pathogenic potential and phylogenetics of Escherichia coli extraintestinal strains isolated in health care units. OBJECTIVE: To establish the genetic diversity of extraintestinal E. coli strains producers of betalactamases TEM, SHV and CTX-M associated with healthcare. MATERIALS AND METHODS: We studied a collection of 12 strains of extraintestinal E. coli with diminished sensitivity to broad-spectrum cephalosporins. Antimicrobial susceptibility was determined by minimum inhibitory concentration. We determined the phylogenetic groups, virulence factors and genes encoding antimicrobial resistance using PCR, and clonal characterization by repetitive element palindromic-PCR rep-PCR. RESULTS: All strains showed resistance to cephalosporins and joint resistance to quinolones and aminoglycosides. The phylogenetic distribution showed that the A and B1 groups were the most frequent, followed by D and B2. We found all the virulence factors analyzed in the B2 group, and fimH gene was the most frequent among them. We found blaCTX-M in all strains,with a higher prevalence of blaCTX-M-8; two of these strains showed coproduction of blaCTX-M-9 and were genetically identified as blaCTXM-65 and blaCTX-M-147 by sequencing. CONCLUSION: The strains under study showed genetic diversity, hosting a variety of virulence genes, as well as antimicrobial resistance with no particular phylogroup prevalence. This is the first report of blaCTX-M alleles in Venezuela and in the world associated to non-genetically related strains isolated in health care units, a situation that deserves attention, as well as the rationalization of antimicrobials use.