Chlamydia psittaci EN AVES PSITACIDAS EN DOS PARQUES ZOOLÓGICOS DE VENEZUELA / Chlamydia psittaci in Psitacines Birds in Two Zoological Parks of Venezuela

RESUMEN La determinación de Chlamydia psittaci (Cp) en aves psitácidas en parques zoológicos de Venezuela representa una estrategia de conservación y preservación para este grupo de aves, donde múltiples especies se encuentran amenazadas de extinción y otras han perdido su capacidad de reincorporación a su hábitat natural. A través de la reacción en cadena de la polimerasa anidada (PCR) fue amplificada la subunidad 16S del ADNr de Cp en 50 muestras de hisopado cloacal de aves psitácidas, reportando una frecuencia de 62 %. El trabajo fue realizado en el Parque Zoológico Las Delicias (PZD) 8 % y el Aquarium de Valencia (AV) 54 %. La elevada frecuencia fue asociada a un genotipo de baja concentración y virulencia debido a la ausencia de signos clínicos de clamidiosis aviar. Estos resultados demuestran la necesidad de promover la detección de Cp, principalmente para el AV que actúa como centro de recepción de ejemplares de decomiso, y, al igual que el PZD, poseen otras especies vulnerables a la extinción con riesgo de infección a Cp.

ABSTRACT The determination of Chlamydia psittaci (Cp) in psittacida birds in zoological parks in Venezuela represents a strategy of conservation and preservation for this group of birds, where multiple species are threatened with extinction and others have lost their capacity of reincorporation to their natural habitat. Through the nested polymerase chain reaction (PCR) the 16S subunit of Cp DNAr was amplified in 50 cloacal swab samples from psittacine birds, reporting a frequency of 62 %. The work was carried out in the Zoo Park Las Delicias (PZD) 8% and the Aquarium of Valencia (AV) 54%. The high frequency was associated with a genotype of low concentration and virulence due to the absence of clinical signs of avian chlamydiosis. These results demonstrate the need to promote the detection of Cp, mainly for the AV that acts as a center of reception of specimens of confiscation, and, like the PZD, have other species vulnerable to extinction with risk of infection to Cp.

La resistina se asocia con la proteína C reactiva y marcadores de riesgo cardiovascular en niños obesos / Resistin is associated with C reactive protein and cardiovascular risk markers in obese children / A resistina associa-se à proteína C reativa e aos marcadores de risco cardiovascular em crianças obesas

El objetivo del trabajo fue evaluar los niveles de resistina sérica y su asociación con la proteína C reactiva (PCR-hs), óxido nítrico (NO) y lípidos plasmáticos (LP) en sujetos de edad pediátrica. Participaron 366 niños y adolescentes (10-16 años), agrupados en eutróficos (n=162) y obesos (n=204). Se les estudió peso, talla, circunferencia de la cintura, resistina sérica, glucosa e insulina basal, LP, NO, PCR-hs, malondialdehído y tensión arterial. Se calculó el IMC, el índice cintura-talla (IC/T) y el HOMA-IR. Se utilizaron las pruebas t de Student, ANOVA, U de Mann y Whitney o Kruskal- Wallis para comparar entre grupos y la correlación de Spearman para determinar asociación entre variables. Los obesos masculinos presentaron niveles superiores de resistina (p<0,05). El género femenino presentó valores más altos de resistina en eutróficos (p=0,012) y con IC/T normal (p=0,011). A mayor concentración de resistina los eutróficos presentaron niveles más altos de triacilglicéridos, pero los obesos mostraron niveles más bajos de triacilglicéridos, HDLc y NO, más altos de PCR-hs y mayor IMC. Los resultados sugieren que la resistina podría ser un factor de riesgo para la enfermedad cardiovascular por su asociación positiva con la PCR-hs e inversa con el NO y la HDLc, parámetros involucrados en la inflamación y la disfunción endotelial.

The aim of this study was to evaluate seric levels of resitin and their association with high-sensitivity C Reactive Protein (hs-CRP), nitric oxide (NO) and plasmatic lipids (PL) in a pediatric age population. A total of 366 children and adolescents (between 10-16 years old) participated, and were grouped into eutrophic (n=162) and obese (n=204). Weight, height, waist circumference, resistin, fasting blood glucose and insulin levels, PL, hs-CRP, NO, malondialdehyde and blood pressure were measured. BMI, waist to height ratio (W/HR) and HOMA-IR were calculated. T-student, ANOVA, Mann-Whitney U-value or Kruskal-Wallis were used to compare between groups and Spearman correlation was used to determine association among variables. Male obese subjects showed higher resistin levels (p<0.05). Female subjects showed higher resistin values in the eutrophic group (p=0.012) and in the normal W/HR (p=0.011). At higher levels of resistin, the eutrophic group showed higher levels of triacylglycerides, but the obese group showed lower triacylglycerides, HDLc and NO levels and higher hs-CRP levels and BMI. These results suggest that resistin could be a risk factor for cardiovascular disease because of its positive association with hs-CRP and inverse association with NO and HDLc, parameters involved in inflammation and endothelial dysfunction.

O objetivo da pesquisa foi avaliar os níveis séricos de resistina sérica e sua associação com a proteína C-reativa (PCR-hs), óxido nítrico (NO) e lipídios plasmáticos (LP) em crianças e adolescentes. O estudo envolveu 366 crianças e adolescentes (10-16 anos), agrupados em eutróficos (n=162) e obesos (n=204). Os sujeitos foram estudados em relação ao peso, altura, circunferência da cintura, resistina sérica, glicose e insulina basal, LP, NO, PCR-hs, malondialdeído e pressão arterial. Os IMC, índice cintura-altura (IC/A) e HOMA-IR foram calculados. Foram utilizados os Testes t de Student, ANOVA, U de Mann e Whitney ou Kruskal-Wallis para comparar entre os grupos e a correlação de Spearman para verificar a associação entre variáveis. Os obesos masculinos mostraram níveis mais elevados de resistina (p<0,05). O sexo feminino apresentou valores mais altos de resistina em eutróficos (p=0,012) e com IC/T normal (p=0,011). À maior concentração de resistina, os eutróficos apresentaram maiores níveis de triacilglicerídeos, mas os obesos apresentaram níveis mais baixos de triacilglicerídeos, HDLc e NO, mais altos de PCR-hs e maior IMC. Os resultados sugerem que a resistina poderia ser um fator de risco para a doença cardiovascular devido à sua associação positiva com a PCR-hs e inversa com o NO e a HDLc, parâmetros envolvidos na inflamação e disfunção endotelial.

[Association between resistin serum levels and dimension and body composition variables in children and adolescents]. / Asociación de la resistina con variables de dimensión y composición corporal en eutróficos y obesos.

BACKGROUND: Resistin is an adipokine secreted in adipose tissue that may be associated with the metabolic and endocrine disorders of obesity. AIM: To assess the association between serum resistin levels and body composition variables, in children and adolescents. MATERIAL AND METHODS: Cross-sectional assessment of 302 subjects, aged 10-16 years. According to body mass index, 124 were classified as eutrophic and 178 as obese. A clinical examination and pubertal assessment were performed. Body weight, height, waist and arm circumferences were measured. Serum resistin levels were measured using an immunoenzymatic assay. RESULTS: Male obese children had significantly higher resistin levels than their eutrophic counterparts. Eutrophic women had higher levels than eutrophic males. No significant association between resistin and pubertal status was observed. In the whole sample and among obese subjects, resistin levels correlated with body mass index, tricipital skinfold, arm circumference, arm fat area and fat mass. CONCLUSIONS: There is a significant association between resistin levels and body composition variables, particularly with indicators of fat accretion.

Asociación de la resistina con variables de dimensión y composición corporal en eutróficos y obesos / Association between resistin serum levels and dimension and body composition variables in children and adolescents

Background: Resistin is an adipokine secreted in adipose tissue that may be associated with the metabolic and endocrine disorders of obesity. Aim: To assess the association between serum resistin levels and body composition variables, in children and adolescents. Material and Methods: Cross-sectional assessment of 302 subjects, aged 10-16 years. According to body mass index, 124 were classified as eutrophic and 178 as obese. A clinical examination and pubertal assessment were performed. Body weight, height, waist and arm circumferences were measured. Serum resistin levels were measured using an immunoenzymatic assay. Results: Male obese children had significantly higher resistin levels than their eutrophic counterparts. Eutrophic women had higher levels than eutrophic males. No significant association between resistin and pubertal status was observed. In the whole sample and among obese subjects, resistin levels correlated with body mass index, tricipital skinfold, arm circumference, arm fat area and fat mass. Conclusions: There is a significant association between resistin levels and body composition variables, particularly with indicators of fat accretion.

Prevalencia de microsporidiosis intestinal en niños con desnutrición severa de un hospital de la ciudad de Maracaibo / Intestinal Microsporidiosis Prevalence in Children With Severe Malnutrition at a Hospital in the City of Maracaibo

Con la finalidad de comparar la prevalencia de Microsporidiosis intestinal en niños con desnutrición severa y niños eutróficos de la Unidad de Recuperación Nutricional del Hospital Chiquinquirá de Maracaibo, Estado Zulia, se realizó la técnica de reacción en cadena de la polimerasa (PCR) para la identificación de Encephalitozoon intestinalis y Enterocytozoon bieneusi en las muestras de heces de 50 niños desnutridos graves y 50 niños eutróficos, cuyas edades fueron estratificadas de la siguiente manera: lactantes menores (0-11 meses), lactantes mayores (12-23 meses), preescolares (2-6 años), y escolares (7 a 12 años). De las especies de microsporidios investigadas, se evidenció Enterocytozoon bieneusi en el 14% de los niños desnutridos graves y 8% en los niños eutróficos, no se obtuvieron amplificaciones para Encephalitozoon intestinalis en los grupos estudiados; al aplicar la prueba estadística X², resultó no significativa para las variables microsporidiosis intestinal y desnutrición. Se concluye que la medida en la que se relacionan la desnutrición, y la microsporidiosis es difícil de esclarecer, ya que la desnutrición es una condición multifactorial y aún no está bien establecido cuál de estos dos factores es la causa y cual la consecuencia.

In order to compare the prevalence of intestinal microsporidiosis in children with severe malnutrition and eutrophic children at the Nutritional Recovery Unit, Chiquinquirá Hospital in Maracaibo, State of Zulia, a chain reaction technique was performed on the polymerase (PCR) to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi in stool samples from 50 healthy and 50 severely malnourished children, whose ages were stratified as follows: younger infants (0-11 months), older infants (12-23 months), preschool (2-6 years) and school children (7-12 years). Of the microsporidia species investigated, Enterocytozoon bieneusi was evident in 14% of severely malnourished children and 8% of the eutrophic children. No amplifications for Encephalitozoon intestinalis were obtained in the groups studied. On applying the chi-square statistical test, the result was not significant for the variables intestinal microsporidiosis and malnutrition. Conclusions are that the extent to which malnutrition and microsporidiosis relate is difficult to clarify, because malnutrition is a multifactorial condition, and it is not yet well established which of these two factors is the cause and which is the consequence.

Detección de Entamoeba histolytica y Entamoeba dispar mediante PCR, en niños menores de cinco años con diarrea, en Maracaibo, Venezuela: Estudio preliminar / Detection of Entamoeba histolytica and Entamoeba dispar by PCR in children, less than five years of age with diarrhea, in Maracaibo, Venezuela: A preliminary study

Para determinar la prevalencia de Entamoeba histolytica como productora de diarrea, se realizó un estudio en niños menores de cinco años con diarrea que asistieron a diversas consultas del Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela. A las muestras de heces obtenidas, se les realizó examen macroscópico, microscópico con solución salina fisiológica 0,85% y lugol, así como coloración de Kinyoun. El resto de la muestra se congeló hasta la extracción del ADN y luego se amplificaron mediante PCR separadas para E. histolytica y E. dispar. De las 50 muestras analizadas mediante examen microscópico, ninguna presentó trofozoítos y/o quistes de Entamoeba histolytica/dispar/moshkovskii, ni coccidios intestinales. Los parásitos detectados fueron Giardia lamblia (6%), Blastocystis sp. (6%), Pentatrichomonas hominis (2%), Ascaris lumbricoides (2%) y Trichuris trichiura (2%). Mediante PCR, 6 muestras (12%) presentaron ADN de E. dispar y 2 (4%) ADN de E. histolytica. Ningún niño presentó asociación de ambas amibas. Los dos niños que presentaron E. histolytica tenían 1 año de edad. E. dispar si fue detectada en niños de menor edad. Se sugiere que la prevalencia de E. histolytica en niños menores de cinco años con diarrea es realmente baja.

To determine the prevalence of Entamoeba histolytica as a producer of diarrhea, a study was conducted in children, less than five years of age, with diarrhea who attended several out patient clinics of the Servicio Autónomo Hospital Universitario, Maracaibo, Venezuela. A macroscopic and microscopic examination with physiological saline, lugol and Kinyoun staining were performed to the stool samples obtained. The remainder of the sample was frozen until DNA extraction, and PCR amplification was performed separately for E. histolytica and E. dispar. Microscopic examination showed no trophozoites and/or cysts of Entamoeba histolytica/dispar/moshkovskii, or intestinal coccidians in any of the 50 samples analyzed. Parasites detected were Giardia lamblia (6%), Blastocystis sp. (4%), Pentatrichomonas hominis (2%), Ascaris lumbricoides (2%) and Trichuris trichiura (2%). By PCR, six samples (12%) had DNA of E. dispar and two (4%) had DNA from E. histolytica; no child showed association of both amoebae. The two children who had E. histolytica were one-year-old. E. dispar was detected in younger children. We suggest that the prevalence of E. histolytica in children under five years is really low.

Evaluación de métodos fenotípicos para la detección de metalobetalactamasas en aislados clínicos de Pseudomonas aeruginosa / Assessment of Phenotypic Methods for Metalobetalactamase Detection in Clinical Isolates of Pseudomonas aeruginosa

Pseudomonas aeruginosa es considerado uno de los patógenos nosocomiales más importantes, siendo común su aislamiento en pacientes de unidades de cuidados intensivos, en quienes además suele presentar multirresistencia, esto limita enormemente las opciones terapéuticas para el tratamiento del paciente, lo que constituye un problema ya que no se disponen de drogas efectivas para el tratamiento. El presente trabajo tiene como objetivo evaluar cuatro métodos fenotípicos para la detección de metalo-beta-lactamasas (método de sinergia del doble disco con EDTA, método del disco combinado de imipenem y meropenem con EDTA, método de E-Test-MBL y el método de Hodge modificado), con un método de referencia (PCR), para determinar el método fenotípico o combinación de estos que sea más valido y seguro para su implementación en los laboratorios de bacteriología de rutina. La sensibilidad y especificidad de los métodos fenotípicos empleados fue buena (más de 90%), de los métodos empleados, el test de Hodge modificado fue el que proporcionó mejores resultados en la detección de aislados de P. aeruginosa productores de MBL (menor tasa de falsos positivos y negativos), al analizar los diferentes métodos combinados para determinar cuál es la asociación con mayor sensibilidad y especificidad, el método del doble disco asociado con el test de Hodge modificado reunieron estas características, estos resultados sugieren que estos dos métodos se deben emplear de rutina en un laboratorio de Bacteriología, por ser fáciles de realizar, económicos, sensibles y específicos.

Pseudomonas aeruginosa is considered one of the most important nosocomial pathogens. Its isolation is commonin intensive care unit patients who also usually have multi-resistance. This greatly limits therapeutic options, since no effective drugs are available for treatment. The study aims to evaluate four phenotypic methods for detecting metallo-beta-lactamases. The double disc synergy with EDTA method, combined imipenem and meropenem disk with EDTA method, the E-Test-MBL method and the modified Hodge method, along with a reference method (PCR), are used to determine the phenotypic method or a combination of these is used that is more valid and safe for implementation in routine bacteriology laboratories. The sensitivity and specificity of the phenotypic methods used was good (over 90%); the modified Hodge test obtained the best results in detecting MBL-producing P. aeruginosa isolates (lower rate of false positive and negative). On analyzing the different combined methods for determining the association with greater sensitivity and specificity, the double disc method associated with the modified Hodge test met these characteristics. Results suggest that these two methods should be routinely used in a bacteriology laboratory, since they are easy to perform, sensitive, specific and economical.

Polimorfismo 3’UTR +62G>A del gen RETN codificante de resistina y asociación con componentes del síndrome metabólico / 3’UTR +62G>A polymorphism of the RETN gene coding resistin and its association with metabolic syndrome components

Fundamento y objetivo: Diversos polimorfismos del gen de la resistina (RETN) se han asociado con alteraciones metabólicas. En este estudio se evaluó la asociación entre el polimorfismo 3’UTR +62G>A del genRETN y elementos del síndrome metabólico. Sujetos y método: La población de estudio (n = 218) fue distribuida en 3 grupos: control, sin alteraciones metabólicas (n = 77); AMnS, con alteraciones metabólicas aisladas (n = 94); y SM, con síndrome metabólico (n = 47). El polimorfismo 3’UTR +62G>A se analizó por PCR-RFLP. Resultados: La obesidad central fue la alteración más frecuentemente descrita tanto en el grupo AMnS (56,4%) como en el SM (91,5%), seguida de bajos niveles de colesterol unido a lipoproteínas de alta densidad (colesterol HDL) (42,6%) en el grupo AMnS e hipertensión arterial (85%) en el grupo SM. La frecuencia del genotipo +62G/A para la población fue del 20,2%. El genotipo G/A fue más frecuente en los grupos SM (38,3%) y AMnS (17%) que en el grupo control (13%). La distribución alélica fue significativamente diferente entre grupos control (+62G = 0,94, +62A = 0,06) y SM (+62G = 0,81, +62A = 0,19) (p = 0,0001). Se encontró asociación significativa entre el genotipo G/A y valores elevados de circunferencia abdominal (p = 0,047), glucemia basal (p = 0,02) y presión arterial sistólica (p = 0,003). Conclusión: Los resultados sugieren una asociación del genotipo RETN +62G/A con valores elevados de presión arterial sistólica, hiperglucemia y obesidad central. La asociación con estas alteraciones parece ser independiente de posibles efectos de agrupación en el contexto de síndrome metabólico (AU)

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International physical activity questionnaire overestimation is ameliorated by individual analysis of the scores.

Sedentarism is considered a risk factor for coronary heart disease and death from any cardiovascular disease. The International Physical Activity Questionnaire (IPAQ) assesses physical activity in metabolic equivalents, using 4 dimensions: occupation, transportation, household activities, and leisure-time physical activity. The purpose of this investigation was to assess physical activity levels in the patients enrolled in the Maracaibo City Metabolic Syndrome Prevalence Study, currently undertaken by the "Dr. Félix Gómez" Endocrine-Metabolic Research Center. Two thousand one hundred eight individuals were recruited and subjected to a standard Medical chart, Graffar scale, and IPAQ long form, applied by trained personnel. Description of the population was done using mean, SD, and coefficient of variation. IPAQ scores were analyzed as medians and distributed by percentiles. From the 2108 individuals, 46.9% were men and 53.1% were women. The most prevalent physical activity was high physical activity with 39.9%, followed by moderate physical activity with 36.9% and low physical activity with 23.2%. Medians for each IPAQ dominion were occupation with 0.00 (0.00-66.00), transportation with 165.00 (0.00-463.00), household activities with 772.50 (0.00-2520.00), and leisure time with 0.00 (0.00-594.00). Using leisure-time scores, a new reclassification was conducted, obtaining 1245 subjects with 0 metabolic equivalents in this dominion. From this new subsample, 43.6% had High physical activity, 56.95% had Moderate physical activity, and 91% had Low physical activity, demonstrating an important overestimation in the former sample results. IPAQ overestimates moderate and vigorous activity in the adult population of the Maracaibo Municipality. Overestimation is mainly located in the household- and gardening-related activity.

[Prevalence of Encephalitozoon intestinalis and Enterocytozoon bieneusi in HIV positive patients to Maracaibo, Venezuela]. / Prevalencia de Encephalitozoon intestinalis y Enterocytozoon bieneusi en pacientes VIH positivos de Maracaibo, Venezuela.

Microsporidioses are considered emerging and opportunistic infections in immunocompromised individuals worldwide. The purpose of this study was to identify the species of intestinal microsporidia in patients with HIV-AIDS from the Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela (SAHUM). Fecal samples were collected from 50 patients with confirmed diagnosis of HIV, during the years 2007 and 2008; the CD4 values were obtained from 42 patients. The samples were analyzed by separate PCRs to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi. Microsporidia species showed a 36% prevalence: ten patients had Encephalitozoon intestinalis, four Enterocytozoon bieneusi and four both species. An inverse and statistically significant relationship between the CD4 count and the presence of microsporidia in the fecal sample was also found. It is remarkable the high prevalence of microsporidia species observed in the HIV patients studied, with a predominance of E. intestinalis.

Single-nucleotide polymorphisms in adiponectin, AdipoR1, and AdipoR2 genes: insulin resistance and type 2 diabetes mellitus candidate genes.

It has already been a decade and a half since the discovery of adiponectin and its role as an insulin sensitizer and only 7 years since its receptors, AdipoR1 and AdipoR2, were described. A single-nucleotide polymorphism (SNP) is a DNA sequence variation that affects only one nucleotide; it may vary from one population to another with different predisposing factors to diseases and other ailments. Once some of the effects of adiponectin and its receptors were known, it was not long until an effort was made to find the associations between specific SNPs of the genes of this hormone and its receptors as genetic risk factors for insulin resistance, type 2 diabetes mellitus, and metabolic syndrome, although these genes were investigated as possible candidates related to the development of these metabolic disorders. All of these possible associations were studied in different populations from France, Finland, the United Kingdom, North America, and Japan, showing hardly concluding results, and because of that it is highly controversial to directly associate one of the genes mentioned above to insulin resistance, type 2 diabetes mellitus, and metabolic syndrome. All of these inconsistencies lead to a review that summarizes the SNPs of the genes of adiponectin, AdipoR1, and AdipoR2 that are mostly related to insulin resistance syndrome, type 2 diabetes mellitus, and metabolic syndrome, although presenting the possible factors that should be taken into account to homogenize the results obtained until now.

Prevalencia de Encephalitozoon intestinalis y Enterocytozoon bieneusi en pacientes VIH positivos de Maracaibo, Venezuela / Prevalence of Encephalitozoon intestinalis and Enterocytozoon bieneusi in HIV positive patients to Maracaibo, Venezuela

Los microsporidios pueden provocar infecciones emergentes y oportunistas en individuos inmunocomprometidos de todo el mundo. Se realizó éste estudio para identificar las especies de microsporidios intestinales presentes en pacientes con VIH-SIDA del Servicio Autónomo Hospital Universitario de Maracaibo (SAHUM). Se recolectaron 50 muestras fecales de individuos con diagnóstico confirmado de VIH durante los años 2007-2008; se obtuvieron las cifras de CD4 de solo 42 pacientes. Las muestras se analizaron mediante PCR separadas para la identificación de Encephalitozoon intestinalis y Enterocytozoon bieneusi. Las especies de microsporidios presentaron un 36% de prevalencia, 10 pacientes presentaron Encephalitozoon intestinalis, 4 Enterocytozoon bieneusi y 4 ambas especies. Se determinó una relación inversamente proporcional y estadísticamente significativa entre el contaje de CD4 y la presencia de microsporidios en la muestra fecal. Es destacable la elevada prevalencia de especies de microsporidios observada en los pacientes VIH estudiados, donde predominó E. intestinalis.

Microsporidioses are considered emerging and opportunistic infections in immunocompromised individuals worldwide. The purpose of this study was to identify the species of intestinal microsporidia in patients with HIV-AIDS from the Servicio Autónomo Hospital Universitario de Maracaibo, Venezuela (SAHUM). Fecal samples were collected from 50 patients with confirmed diagnosis of HIV, during the years 2007 and 2008; the CD4 values were obtained from 42 patients. The samples were analyzed by separate PCRs to identify Encephalitozoon intestinalis and Enterocytozoon bieneusi. Microsporidia species showed a 36% prevalence: ten patients had Encephalitozoon intestinalis, four Enterocytozoon bieneusi and four both species. An inverse and statistically significant relationship between the CD4 count and the presence of microsporidia in the fecal sample was also found. It is remarkable the high prevalence of microsporidia species observed in the HIV patients studied, with a predominance of E. intestinalis.

[Detection of Entamoeba histolytica and Entamoeba dispar by PCR in children, less than five years of age with diarrhea, in Maracaibo, Venezuela. A preliminary study]. / Detección de Entamoeba histolytica y Entamoeba dispar mediante PCR, en niños menores de cinco años con diarrea, en Maracaibo, Venezuela. Estudio preliminar.

To determine the prevalence of Entamoeba histolytica as a producer of diarrhea, a study was conducted in children, less than five years of age, with diarrhea who attended several out patient clinics of the Servicio Aut6nomo Hospital Universitario, Maracaibo, Venezuela. A macroscopic and microscopic examination with physiological saline, lugol and Kinyoun staining were performed to the stool samples obtained. The remainder of the sample was frozen until DNA extraction, and PCR amplification was performed separately for E. histolytica and E. dispar. Microscopic examination showed no trophozoites and/or cysts of Entamoeba histolytica/dispar/moshkovskii, or intestinal coccidians in any of the 50 samples analyzed. Parasites detected were Giardia lamblia (6%), Blastocystis sp. (4%), Pentatrichomonas hominis (2%), Ascaris lumbricoides (2%) and Trichuris trichiura (2%). By PCR, six samples (12%) had DNA of E. dispar and two (4%) had DNA from E. histolytica; no child showed association of both amoebae. The two children who had E. histolytica were one-year-old. E. dispar was detected in younger children. We suggest that the prevalence of E. histolytica in children under five years is really low.

[3'UTR +62G>A polymorphism of the RETN gene coding resistin and its association with metabolic syndrome components]. / Polimorfismo 3'UTR +62G>A del gen RETN codificante de resistina y asociación con componentes del síndrome metabólico.

BACKGROUND AND OBJECTIVE: The polymorphism of the resistin gene (RETN/RSTN) has been associated with metabolic alterations. In this study the association between the 3'UTR +62G>A polymorphism and metabolic syndrome components was evaluated. SUBJECTS AND METHODS: The population (n=218) was distributed in 3 groups: the control group with no metabolic alterations (n=77), nSMA group with isolated metabolic alterations (n=94) and MS group with metabolic syndrome (n=47). The 3'UTR +62G>A polymorphism was analyzed by PCR-RFLP. RESULTS: Central obesity was the most frequent alteration in both nSMA (56.4%) and MS (91.5%) groups followed by low c-HDL levels in the nSMA group (42.6%) and arterial hypertension in the MS group (85%). The frequency of the +62G/A genotype was 20.2% in the population. The G/A genotype was more frequently found in the MS (38.3%) and nSMA (17%) groups than in the control group (13%). The allelic distribution between the control group (+62G=0.94, +62A=0.06) and MS group (+62G=0.81, +62A=0.19) was significantly different (P=.0001). Significant associations between the G/A genotype and high values of abdominal circumference (P=.047), basal glycemia (P=.02) and systolic arterial pressure (P=.003) were found. CONCLUSION: The findings suggest the association between the G/A genotype and high values of systolic arterial pressure, basal glycemia and abdominal circumference. This association was independent of the metabolic syndrome context.

Evidence of zoonotic Chlamydophila psittaci transmission in a population at risk in Zulia state, Venezuela.

OBJECTIVE: This study was aimed at investigating the frequency of infection by Cp. psittaci and determining its genotype in individuals at potential risk of exposure to the bacteria. METHODOLOGY: The study involved 170 individuals: a risk group (n= 96) and a low-risk control group (n=74). Cp. psittaci was detected and genotyped by single-tube nested PCR and ompA gene sequencing. RESULTS: Eight (8.3 %) positive cases were detected in the risk group and 1 (1.4 %) in the control group (p<0.04). Cp. psittaci was found in 16.7 % of pigeons' fecal samples. Cp. psittaci infection with was more frequent in symptomatic (17.7 %) than asymptomatic (6.3 %) individuals in the risk group. Analysing the genomes isolated from human and bird specimens revealed the presence of genotype B. CONCLUSION: The presence of Cp. psittaci genotype B in the population being evaluated could have been attributed to zoonotic transmission from pigeons to humans, an underestimated potential public health problem in Venezuela requiring the health authorities' involvement.

Detección de la mutación S315T en el gen katg como estrategia para identificación de Mycobacterium tuberculosis resistente a isoniacida en un laboratorio de referencia / Detection of S315T mutation in the katg gene as a strategy for identification of isoniazid-resistant Mycobacterium tuberculosis in a reference laboratory

Introduction: The emergence of strains of Mycobacterium tuberculosis resistant to drugs is a public health problem. Aim: To characterize the resistance to isoniazid (INH) in M. tuberculosis. Methods: Phenotypic and genotypic methods were used to determine the contribution of mutations at 315 codon of katG gene to the phe-notypic expression of resistance. The analysis of susceptibility to antibiotics was performed by the proportional method of Canetti and nitrate reductase method.Genotypic analysis of INH resistance was performed by PCR-RFLP. Results: 193 strains of M. tuberculosis from patients with respiratory symptoms were analyzed. Nineteen (9.8%) strains resistant to INH were identified, of which 12 (63.2%) showed resistance to other drugs. Genotypic analysis allowed to detect the mutation S315T in the katG gene in 15 of 17 strains phenotypically resistant to INH, showing a sensitivity of 88.24%, 100% specificity, 100% positive predictive value, 92% negative predictive value and high concordance with phenotypic methods (kappa = 0.85 (p < 0.01). Conclusion: The S315T mutation in the katG gene is the predominant mechanism of INH resistance in our circulating strains. This feature could be used as a rapid diagnostic tool with potential to detect at least 88% of isoniazid resistant strains, with great impact on the therapeutic management of patients.

Introducción: La emergencia de cepas de Mycobacterium tuberculosis multi-resistentes a antimicrobianos constituye un problema de salud pública. Objetivo: Caracterizar la resistencia a isoniacida (HIN) en cepas de M. tuberculosis. Métodos: Se aplicaron métodos fenotí-picos y genotípicos para determinar la contribución de mutaciones en el codón 315 del gen katG a la expresión fenotípica de resistencia. El análisis fenotípico de susceptibilidad a antimicrobianos se realizó por métodos de las proporciones de Canetti y nitrato reductasa. El análisis genotípico se realizó por RPC-PLFR. Resultados: Se analizaron 193 cepas de M. tuberculosis aisladas de pacientes sintomáticos respiratorios. Se identificaron 19 (9,8%) cepas resistentes a HIN, de las cuales, 12 (63,2%) exhibieron resistencia a otros antimicrobianos. El análisis genotípico permitió detectar la mutación katG S315T en 15/17 cepas fenotípicamente resistentes a HIN, exhibiendo una sensibilidad de 88,24%, especificidad de 100%, valor predictor positivo de 100%, valor predictor negativo de 92% y alta concordancia al comparar con los métodos fenotípicos (kappa = 0,85 (p < 0,01). Conclusión: La mutación S315T en el gen katG representa un mecanismo de desarrollo de resistencia a HIN predominante en las cepas circulantes en nuestro medio, característica que podría ser utilizada como herramienta diagnóstica rápida para detectar al menos 88% de las cepas resistentes a isoniacida, con gran impacto en el manejo terapéutico de los pacientes.

Detección fenotípica de metalobetalactamasas en aislados clínicos de Pseudomonas aeruginosa / Phenotypic detection of metallo-beta-lactamase in clinical isolates of Pseudomonas aeruginosa

Pseudomonas aeruginosa, es considerado uno de los más importantes gérmenes hospitalarios, siendo común su aislamiento en pacientes hospitalizados, adicionalmente, este microorganismo presenta una marcada multiresistencia, lo que incrementa la mortalidad. El tratamiento de estos pacientes suele ser difícil, ya que además de su resistencia natural, Pseudomonas puede adquirir mecanismos de resistencia para prácticamente la totalidad de los antimicrobianos disponibles para su tratamiento, por lo que cada vez es más frecuente y necesario el empleo de antibióticos como los carbapenems, lo que facilita la adquisición de mecanismos de resistencia a estas drogas. El presente estudio intenta determinar la producción de metalobetalactamasas (MBL) en aislados clínicos de Pseudomonas aeruginosa, utilizando para ello dos métodos fenotípicos. Se utilizó el método del doble disco (MDD) y el test de Hodge modificado (MHT). Se analizaron 726 aislados clínicos de P. aeruginosa, el 20,11% (146) de estos fueron resistentes a imipenem (IPM) y meropenem (MEM), por lo que se les realizaron los dos métodos fenotípicos, de los 146 aislados resistentes a carbapenems, 139 fueron positivas para el MDD, mientras que 144 lo fueron para el MHT, estos dos métodos permitieron confirmar la presencia de una carbapenemasa tipo MBL en el 98,63% de los aislados de P. aeruginosa, por otra parte, cinco aislados no fueron positivos para el MDD pero si para el MHT, lo que indicaría la presencia de carbapenemasas no MBL en estos aislado, también se obtienen 2 aislados que a pesar de ser IPM y MEM resistentes fueron negativos por los dos métodos fenotípicos utilizados, esto indicaría la presencia de un mecanismo de resistencia no enzimático que confiere resistencia a carbapenems...

Pseudomonas aeruginosa is considered one of the most important hospital germs; its isolation is common in hospitalized patients. In addition, this microorganism has a marked multi-resistance, which increases mortality. Treatment of these patients is often difficult, since in addition to its natural resistance, Pseudomonas can obtain resistance mechanisms to virtually all antimicrobial drugs available for its treatment; due to this, its appearance is increasingly frequent and necessitates the use of antibiotics such as carbapenems, which facilitates the acquisition of resistance mechanisms to these drugs. This study attempts to determine the production of metallo-beta-lactamase (MBL) in clinical isolates of Pseudomonas aeruginosa, utilizing two phenotypic methods: the double disc method (MDD) and the modified Hodge test (MHT). 726 clinical isolates of P. aeruginosa were analyzed; 20.11% (146) of these were resistant to imipenem (IPM) and meropenem (MEM); 139 were positive for the MDD, while 144 were positive for the MHT. These two methods permitted confirming the presence of an MBL-type carbapenemase in 98.63% of P. aeruginosa isolates; five isolates were negative for the MDD but positive for the MHT, indicating the presence of non-MBL-type carbapenemase in these isolates. Also, 2 isolates were obtained that, despite being resistant to IPM and MEM, were negative according to the two phenotypic methods used; this would indicate the presence of a non-enzymatic resistance mechanism conferring resistance to carbapenems. The use of phenotypic methods for detecting MBL in P. aeruginosa isolates is quite an acceptable option for use in routine laboratories where specialized molecular biology tests are not available.

Polimorfismos del gen UCP-3 en individuos con Síndrome Metabólico del Municipio Maracaibo-Venezuela

Objetivo: La frecuencia de Síndrome Metabólico (SM) en poblaciones adultas está en constante incremento en el Estado Zulia. En este estudio se determinó la frecuencia de los polimorfismos G304A exón 3 del gen UCP-3 y su asociación con los componentes del SM y composición corporal. Métodos: Se estudiaron 92 individuos (45 con diagnóstico de SM, según los criterios de la Federación Internacional de Diabetes y 47 individuos sanos). Se realizaron las determinaciones antropométricas, se tomo la presión arterial y en ayunas se cuantificaron los parámetros bioquímicos. Las versiones polimórficas fueron analizadas por PCR-RFLP. Resultados: La frecuencia genotípica de G/G y G/A fue de 84,45 y 15,55 respectivamente en el grupo con SM con una frecuencia alélica G y A de 0,92 y 0,08 respectivamente. En el grupo testigo la frecuencia genotípica de G/G y G/A fue de 97,88 y 2,12 respectivamente, con una frecuencia alélica G y A de 0,99 y 0,01. Se observaron diferencias significativas entre genotipo G/G y G/A en valores de glucosa basal (p<0,0001), tensión arterial sistólica (p<0,004), triacilgicéridos (p<0,042) en pacientes con SM y porcentaje de grasa (p<0,04). También se observó diferencias significativas en valores de porcentaje de grasa (p< 0,043), VLDL-c (p< 0,04), LDL-c (p<0,019) y triacilglicéridos en sexo femenino con SM. Conclusión: En la muestra estudiada se observó asociación aparente entre el genotipo G/A del gen UCP-3 con hiperglicemia, hipertensiòn arterial sistólica e incremento del porcentaje de grasa así como dislipidemia en el caso particular del sexo femenino.

Objectives: The frequency of metabolic syndrome (MS) in adult populations is constantly increasing in Zulia State. In this study we determined the frequency of polymorphisms G304A exon 3 of the UCP-3 gene and its association with metabolic syndrome components and body composition. Methods: The sample consisted of 92 individuals (45 with diagnosis of MS, according to the criteria of the International Diabetes Federationand 47 healthy subjects). versions were analyzed by PCR-RFLP. Results: The genotype frequency of G/G and G/A was 84.45 and 15.55 respectively in the MS group with a G and A allele frequency of 0.92 and 0.08 respectively, while the genotype frequency in the control group of G/G and G/A was 97.88 and 2.12 respectively with a G and A allele frequency of 0.99 and 0.01. Significant differences were observed between genotype G/G and G/A fasting glucose values (p <0.0001), systolic blood pressure (p<0.004), triacilgicéridos (p <0.042) in patients with MS and fat percentage (p <0.04). Also observed significant differences in fat percentage values (p <0.043), VLDL-c (p <0.04), LDL-c (p <0.019) and triglycerides in females with MS. Conclusion: In the sample studied was observed apparent association between the genotype G/A UCP-3 gene with hyperglycemia, hypertension and increased systolic blood fat percentage and dyslipidemia in the particular case of female.

[Detection of S315T mutation in the katg gene as a strategy for identification of isoniazid-resistant Mycobacterium tuberculosis in a reference laboratory]. / Detección de la mutación S315T en el gen katg como estrategia para identificación de Mycobacterium tuberculosis resistente a isoniacida en un laboratorio de referencia.

INTRODUCTION: The emergence of strains of Mycobacterium tuberculosis resistant to drugs is a public health problem. AIM: To characterize the resistance to isoniazid (INH) in M. tuberculosis. METHODS: Phenotypic and genotypic methods were used to determine the contribution of mutations at 315 codon of katG gene to the phenotypic expression of resistance. The analysis of susceptibility to antibiotics was performed by the proportional method of Canetti and nitrate reductase method.Genotypic analysis of INH resistance was performed by PCR-RFLP. RESULTS: 193 strains of M. tuberculosis from patients with respiratory symptoms were analyzed. Nineteen (9.8%) strains resistant to INH were identified, of which 12 (63.2%) showed resistance to other drugs. Genotypic analysis allowed to detect the mutation S315T in the katG gene in 15 of 17 strains phenotypically resistant to INH, showing a sensitivity of 88.24%, 100% specificity, 100% positive predictive value, 92% negative predictive value and high concordance with phenotypic methods (kappa = 0.85 (p < 0.01). CONCLUSION: The S315T mutation in the katG gene is the predominant mechanism of INH resistance in our circulating strains. This feature could be used as a rapid diagnostic tool with potential to detect at least 88% of isoniazid resistant strains, with great impact on the therapeutic management of patients.

Curable sexually transmitted infections among female sex workers in a population of Zulia State, Venezuela / Infecciones de transmisión sexual curables en trabajadoras sexuales en una población del estado Zulia, Venezuela

Sexually transmitted infections (STIs) represent a public health problem worldwide. The aim of this study was to investigate the prevalence of curable STIs caused by Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum in female sex workers in a population from Zulia State, Venezuela. Seventy eight (78) women attended a health monitoring sanitary controls were evaluated, and PCR amplification assays were used to detect the three microorganisms in endocervical samples. In 33.3% of the samples, at least one microorganism was detected: U. urealyticum was found more frequently (25.6%), followed by N. gonorrhoeae (18%), and C. trachomatis (12.8%). A significant association between N. gonorrhoeae and C. trachomatis was found (p <0.0001). STIs cases represented 42.9% and 28% for symptomatic and asymptomatic groups, respectively. In the symptomatic group, N. gonorrhoeae was 2.4 times (28.6%) more frequent than in the asymptomatic one (12%) (p=0.015), particularly associated with mucopurulent discharge (p=0.025). No association was found between C. trachomatis (p=0.078), and U. urealyticum (p=0.432) with clinical manifestations. Prevalence of curable STIs in the study population was relatively low compared with other high-risk populations worldwide. The results support the possible association between C. trachomatis and N. gonorrhoeae, therefore, treatment would be indicated against both pathogens when one of them is detected in vulnerable populations.

Las infecciones de transmisión sexual (ITS) representan un problema de salud pública a nivel mundial. El objetivo de este estudio fue investigar la prevalencia de ITS curables causadas por Neisseria gonorrhoeae, Chlamydia trachomatis y Ureaplasma urealyticum en trabajadoras sexuales de una población del estado Zulia, Venezuela. Se evaluaron 78 mujeres que asistieron a jornadas de control sanitario y se utilizaron ensayos de amplificación por PCR para detectar los tres microorganismos en muestras endocervicales. En 33,3% de las muestras, se detectó al menos un microorganismo: U. urealyticum fue encontrado con mayor frecuencia (25,6%), seguido de N. gonorrhoeae (18%) y C. trachomatis (12,8%). Se encontró asociación significativa entre N. gonorrhoeae y C. trachomatis (p<0,0001). Los casos de ITS representaron porcentajes de 42% y 28% para los grupos sintomático y asintomático, respectivamente. N. gonorrhoeae fue 2,4 veces más frecuente en el grupo sintomático (28,6%) que en el asintomático (12%) (p=0,015), particularmente asociado con secreción mucopurulenta (p=0,025). No se encontró asociación entre C. trachomatis (p=0,078) y U. urealyticum (p=0,432) con manifestaciones clínicas. La prevalencia de ITS de la población en estudio fue relativamente baja comparada con otras poblaciones de alto riesgo a nivel mundial. Los resultados apoyan la posible asociación entre C. trachomatis y N. gonorrhoeae, por lo tanto se debería considerar el tratamiento contra ambos patógenos, cuando uno de ellos sea detectado en poblaciones vulnerables.