Effectiveness of non-immersive virtual reality in the management of procedure-related pain in preschool children: a randomized clinical trial.

The objective of this study is to assess the effectiveness of non-immersive virtual reality as a pain-distraction measure in children between the ages of 3 and 5 years undergoing painful injection procedures in an outpatient setting. We carried out a randomized, unmasked clinical trial in children undergoing venipuncture or intramuscular injection procedures. Patients were randomized to a distraction virtual reality video or standard care. After the procedure, three independent observers (parents, researchers, nursing staff) rated pain on the LLANTO pain scale. We recruited 122 subjects, half of which were randomized to virtual reality. The median age was of approximately 60 months (IQR: 15 months), and the sample was balanced with regard to sex. There were significant differences in LLANTO scales scores between the VR subjects and controls of - 3.34 (95% CI - 4.15; - 2.54), - 3.02 (95% CI - 3.90; - 2.14), and - 2.98 (95% CI - 3.87; - 2.09), as rated by parents, researchers, and nursing staff, respectively. Agreement between raters was high for all three types of observers, with Cohen Kappas over 0.79 in all cases. Bivariate analysis showed reductions in the risk of obtaining higher scores in the LLANTO scale. Linear regression models showed a reduction of approximately 3 points in the scale, regardless of the type of observer. These models were adjusted for sex, age, kind of procedure, use of prior analgesia, and recruitment center. CONCLUSIONS: Non-immersive virtual reality is an effective adjunctive therapy for the reduction of pain in children undergoing painful injection procedures in an outpatient setting. This strategy may be used to improve the quality of care in pediatric outpatient services. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03985930 (Registered June 14, 2019). WHAT IS KNOWN: •The use of immersive virtual reality (VR) has been described as an effective adjunctive distraction method during painful procedures in children over 5 years. WHAT IS NEW: •The utility of non-immersive VR in children below that age is not yet clear. This randomized clinical trial comparing non-immersive VR vs. standard care showed an average reduction of three points in the LLANTO pain scale favoring non-immersive VR. Non-immersive VR is an effective and inexpensive non-pharmacological technique that reduces fear and pain in pediatric patients.

Self-limited epilepsy with autonomic seizures: A case report.

Self-limited epilepsy with autonomic seizures, formerly known as benign occipital epilepsy of childhood or Panayiotopoulos syndrome is a focal epilepsy that is part of the epileptic syndromes with onset during childhood. The objective of this report is to raise awareness about its importance and describe the clinical manifestations, timely diagnosis, and treatment. A pediatric patient admitted with gastrointestinal manifestations is presented. The autonomic manifestations must be considered as part of the clinical spectrum that includes this disease and the digestive and autonomic manifestations that mask the diagnosis, sometimes even in the absence of motor seizures themselves. Electroencephalographic confirmation was performed, avoiding cataloging it in other differential diagnoses.

Probióticos: una mirada al mecanismo de acción y aplicaciones clínicas en Pediatría

Introducción: Los probióticos son microorganismos vivos que brindan beneficios al huésped mediante diversos mecanismos de acción. Han sido fuente de estudio en diversas patologías pediátricas, mostrando algunos resultados prometedores. Objetivo: Elaborar una revisión de la literatura sobre los mecanismos de acción y la evidencia actual que tienen los probióticos sobre la salud infantil. Materiales y métodos: Se realizó una revisión narrativa de la literatura con estrategia de búsqueda sistemática de la literatura con términos MESH acerca de los mecanismos de acción de los probióticos y su uso. Se incluyeron metaanálisis, revisiones sistemáticas y ensayos clínicos aleatorizados. Resultados: Los probióticos son una nueva herramienta terapéutica usada para mejorar la salud infantil. Se ha encontrado efecto benéfico en diarrea, en enterocolitis necrosante con una disminución significativa de la mortalidad y se ha mostrado evidencia significativa en las horas de llanto en cólico del lactante. Conclusión: Se requieren más estudios en otro tipo de enfermedades como estreñimiento y en algunos procesos alérgicos e inflamatorios. Los ensayos revisados ofrecen un panorama prometedor, pero la elección de un probiótico debe ser personalizado de acuerdo con la edad, enfermedad, cepa y dosis, dado que cada uno de ellos tiene múltiples mecanismos de acción que impactan de manera diferente en la eficacia clínica.

Introduction: Prebiotics are living microorganisms that provide benefits to the host through various mechanisms of action. They have been a source of study in various pediatric pathologies showing some promising results. Objective: To prepare a review on the mechanisms of action and current evidence that prebiotics have on child health. Materials and methods: A narrative review of the literature was carried out with a systematic literature search strategy with MESH terms about the mechanisms of action of probiotics and their use. Meta-analyzes, systematic reviews, and randomized clinical trials were included. Results: Probiotics are a new therapeutic tool used to improve children's health. A beneficial effect has been found in diarrhea, in necrotizing enterocolitis with a significant decrease in mortality and significant evidence has been shown in the hours of crying in colic in infants. Conclusion: The trials reviewed offer a promising picture, but the choice of a probiotic must be customized according to the age, disease, bacterial strain and dose, since each one has different action mechanisms and clinical effectiveness. More studies are required in some allergic and inflammatory diseases.

Factors associated with self-medication of antibiotics by caregivers in pediatric patients attending the emergency department: a case-control study.

INTRODUCTION: Antibiotic self-medication is a common practice in pediatric caregivers in low-income countries with limited resources and represents a public health problem. Our study sought to determine what factors are associated with this practice, including differences in knowledge or attitudes of caregivers who attend a pediatric emergency service. METHODS: Case-control study based on surveys of caregivers of pediatric patients brought to the emergency room with clinical symptoms suggestive of acute infection. Cases were defined as those caregivers who reported self-medication of antibiotics for the current illness and controls where those who did not report self-medication. Information was collected through a self-administered questionnaire that inquired about demographic and family characteristics, attitudes and knowledge toward self-medication of antibiotics. Data were compared using logistic regression and are presented with odd ratios and confidence intervals. RESULTS: A total of 728 caregivers, 182 cases and 546 controls were included. We found that higher parental education, both in mothers (OR 0.56, 95% CI 0.40-0.79) and fathers (OR 0.62, 95% CI 0.43-0.89) was associated with less self-medication. Attitudes such as always requesting antibiotics from their doctors (OR 3.92, 95% CI 1.59-9.66), frequently buying antibiotics without a prescription (OR 23.66, 95% CI 11.76-47.59) and giving advice on antibiotics among family members (OR 2.90, 95% CI 1.75-4.82) resulted in an increased likelihood of self-medication. There was also a higher probability of antibiotic self-medication in older children (OR 1.13, 95% CI 1.09-1.17), those with a greater number of siblings (OR 1.25, 95% CI 1.09-1.43) and in those cases that received antibiotics within the last 3 months (OR 6.27, 95% CI 4.35-9.04). Overall knowledge of risk of antibiotic self-medication was low. CONCLUSIONS: Some patient and family characteristics such as age, number of siblings, recent antibiotic usage and inappropriate attitudes are strongly related to antibiotic self-medication. These findings will inform future interventions to reduce self-medication in children.

Differences Between Methicillin-susceptible Versus Methicillin-resistant Staphylococcus aureus Infections in Pediatrics: Multicenter Cohort Study Conducted in Bogotá, Colombia, 2014-2018.

BACKGROUND: The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) has changed in recent years. The present article is intended to establish differences between clinical, laboratory and imaging findings and outcomes of MSSA and MRSA infections, as well as among subgroups of infection such as skin and soft tissue infection, osteoarticular, bacteremia or pneumonia in a pediatric population from Bogota, Colombia. METHODS: Retrospective cohort study using clinical records of patients under 18 years of age treated at the participating centers in Bogota, Colombia, between 2014 and 2018. The first positive S. aureus culture was studied. MSSA and MRSA were compared. The χ2 test, Fisher exact test, and Kruskal-Wallis test were calculated, and the statistical significance was presented using the difference and its 95% CI. RESULTS: Five hundred fifty-one patients were included; 211 (38%) corresponded to MRSA and 340 (62%) to MSSA for a total of 703 cultures. A significantly higher probability of having an MSSA infection than MRSA was found in patients with previous heart disease (3.3% vs. 0.5%), neurologic disease (5.9% vs. 2.5%), recent major surgeries (11% vs. 5%) or who has an implanted device (11% vs. 4%). In contrast, in severe MRSA infections (bacteremia, osteoarticular infections and pneumonia), a higher rate of complications was seen (admission to the pediatric intensive care unit, mechanical ventilation and vasoactive support), and in osteoarticular MRSA, more than 1 surgery per case was seen (89% vs. 61%). Laboratory results and mortality were similar. CONCLUSIONS: MRSA was associated with a more severe course in bacteremia, osteoarticular infections and pneumonia. Some classical risk factors associated with MRSA infections were found to be related to MSSA. In general, with the exception of skin and soft tissue infection, there was an increased risk of pediatric intensive care unit admission and mechanical and inotropic support with MRSA in a pediatric population.

Use of the ReSVinet Scale for parents and healthcare workers in a paediatric emergency service: a prospective study.

Introduction: Most scales for acute respiratory infection (ARI) are limited to healthcare worker (HCW) use for clinical decision-making. The Respiratory Syncytial Virus network (ReSVinet) Scale offers a version for parents that could potentially help as an early warning system. Objective: To determine whether or not the ReSVinet Scale for ARI in infants can be reliably used by HCWs and parents in an emergency service. Methods: A prospective study was done of infants with ARI who were admitted to a paediatric emergency room to assess the ReSVinet Scale when used by faculty (paediatric doctor-professors), residents (doctors doing their first specialty in paediatrics) and parents. Spearman's correlation and a weighted kappa coefficient were used to measure interobserver agreement. Internal consistency was also tested by Cronbach's alpha test. Results: Overall, 188 patients, 58% male, were enrolled. A Spearman's correlation of 0.92 for faculty and resident scoring and 0.64 for faculty or resident and parent scoring was found. The weighted kappa coefficients were 0.78 for faculty versus residents, 0.41 for faculty versus parents, and 0.41 for residents versus parents. Cronbach's alpha test was 0.67 for faculty, 0.62 for residents and 0.69 for parents. Conclusion: There was good correlation in the ReSVinet scores between health professionals when used in the paediatric emergency area. Agreement between parents and health professionals was found to be more variable. Future studies should focus on finding ways to improve its reliability when used by parents before the scale is used in the emergency room.

Extended-Spectrum b-Lactamase-Producing Enterobacteriaceae Causing Community-Acquired Urinary Tract Infections in Children in Colombia.

OBJECTIVE: To characterize the pediatric patients presenting at the two pediatric centers in Bogotá, with first isolate urine culture of community-acquired extended-spectrum b-lactamase (ESBL)-producing enterobacteriaceae. METHODS: Review of microbiological data of children between January, 2012 and December, 2018, obtained using the WHONET software. RESULTS: A total of 2657 Escherichia coli, Klebsiella spp and Proteus mirabilis - positive urine cultures were obtained within a 6-year period; data of 132 patients were finally selected. Frequency of ESBL-producing bacteria infections in community-acquired urinary tract infections (UTI) was 5%: 123 E. coli (93.2%), 7 K. pneumoniae (5.2%), 1 K. oxytoca (0.8%), and 1 P. mirabilis (0.8%). CONCLUSION: A predominance of female sex, preschool children, and lower tract urinary infections were found, as well as a low frequency of comorbidities. Adequate sensitivity to amikacin and nitrofurantoin was found in this study.

Microbiota intestinal en pediatría / Intestinal microbiota in the pediatric population

La microbiota intestinal es el conjunto de millones de microrganismos vivos ubicados en el tracto gastrointestinal. Es indispensable en múltiples funciones del organismo, regulación de la inmunidad, en aspectos nutricionales y procesos de inflamación sistémica entre otros. La disbiosis es la alteración del equilibrio de la microbiota normal, debido a cambios en la composición, funcionamiento, orden o su distribución; esto puede predisponer al individuo a la adquisición de enfermedades gastrointestinales, alérgicas y metabólicas, entre otras. El objetivo del presente artículo es realizar una revisión narrativa de la literatura sobre los conceptos claves de la microbiota intestinal, sus asociaciones fisiopatológicas con desórdenes gastrointestinales, alérgicos y metabólicos en pediatría.

ntestinal microbiota are the millions of living microbial communities that inhabit the gastrointestinal tract. It is essential for multiple functions of the human organism, such as, immune-regulation, in nutritional aspects, and systemic inflammatory processes, among others. Dysbiosis refers to the alteration of the equilibrium of normal microbiota due to shifts in its composition, functioning, order or distribution; this can predispose the individual to develop gastrointestinal, allergic and metabolic diseases among others. The aim of this article was to conduct a narrative review of the literature on the key concepts of intestinal microbiota, and its pathophysiological associations with gastrointestinal, allergic and metabolic disorders in pediatrics.

Difference in the duration of pediatric diabetic ketoacidosis: Comparison of new-onset to known type 1 diabetes.

OBJECTIVE: To compare the duration (hours until HCO3- ≥ 15 mmol/L) of diabetic ketoacidosis (DKA) episodes that are the first manifestation of new type 1 diabetes (NT1D) and those that are a complication in patients with previously diagnosed type 1 diabetes (PT1D). METHODS: A multicenter retrospective cohort study was designed. The duration of DKA was measured from the start of the treatment. The primary outcome was the comparison of the time needed in each group to reach HCO3- ≥ 15 mmol/L. The secondary outcomes were the comparison of the time to reach pH ≥ 7.3 and length of hospital stay in each group. Data were analyzed with a bivariate analysis of the variables vs primary outcome. Then, a regression model was analyzed. Results There were 305 episodes included (NT1D: 115, PT1D: 190). DKA in the NT1D group lasted longer (NT1D 20 (16-19) vs PT1D 12 (8-16), hours, P < .01) with a significant difference in each level of DKA severity. This group also took longer to reach pH ≥ 7.3 (NT1D 16 (12-22) vs PT1D 9 (6-12), hours, P < .01) and had a longer hospital stay (NT1D 9 (6-12) vs PT1D 7 (4-10), hours, P < .01). CONCLUSION: The duration of DKA is longer in patients with NT1D regardless of characteristics like DKA severity, duration of symptoms, and type of treatments received.

Lemierre's Syndrome Caused by Streptococcus anginosus Presenting as Postseptal Cellulitis in a Pediatric Patient.

Lemierre's syndrome is an infrequent disease characterized by septic thrombosis of the internal jugular vein followed by pulmonary embolism generally occurring after upper respiratory and gastrointestinal tract infections. We present the case of a 15-year-old female patient with postseptal cellulitis and cervical abscess who developed pulmonary embolism and pleural effusion secondary to internal jugular vein thrombosis. Cultures were positive for Streptococcus anginosus, antibiotic treatment was established with satisfactory clinical outcome. High clinical suspicion is required for a diagnosis. The mainstay of treatment is a multidisciplinary approach based on two essential pillars: antibiotic therapy and surgical drainage. This is an important case because of the unusual presentation, the isolation of an infrequent pathogen, and the primary infection site (postseptal cellulitis), which are rare characteristics of this condition in the pediatric population.

Alteraciones urinarias en niños con primera infección urinaria e infección urinaria recurrente / Urinary disorders in children with first urinary infection and recurrent urinary tract infection

Introducción: las infecciones del tracto urinario son un problema frecuente en la población pediátrica. La infección del tracto urinario recurrente tiene mayor riesgo de cicatrices renales y daño glomerular. Objetivo: describir la frecuencia de hematuria, presencia de proteínas en orina, elevación en tensión arterial y hallazgos ecográficos en pacientes pediátricos desde 1 mes hasta 14 años de edad, con diagnóstico de infección del tracto urinario. Métodos: estudio descriptivo transversal, realizado entre octubre de 2014 y febrero de 2016, en pacientes que asistieron al servicio de urgencias pediátricas del Hospital de San José, con diagnóstico de infección del tracto urinario. Resultados: se diagnosticaron 125 pacientes con infección del tracto urinario, la mediana de edad fue de 2,75 años, más frecuente en mujeres (75,2 por ciento) y la mayoría recibió tratamiento intrahospitalario (58,4 por ciento). Se observó que en el grupo de infección del tracto urinario recurrente fue más frecuente el hallazgo de hematuria, que en la infección del tracto urinario primer episodio (21,9 vs. 11,8 por ciento), al igual que las malformaciones renales (32,0 vs. 14,5 %). En la tensión arterial no se observaron grandes diferencias entre los grupos, con una frecuencia de tensión arterial elevada en hospitalizados de 19,6 por ciento para primer episodio, y 18,2 por ciento en recurrentes. La presencia anormal de proteínas en orina fue de 35,4 por ciento en el grupo de infección del tracto urinario primer episodio. Conclusiones: son frecuentes la presencia de hematuria, malformaciones renales y elevación de la tensión arterial en pacientes con infección del tracto urinario. Se encuentra una frecuencia inusual de pacientes con elevación de la presión arterial y proteínas en orina, probablemente por falsos positivos. Se debe insistir en seguimiento ambulatorio del uroanálisis y la tensión arterial, para garantizar que esta se normalice o para detectar daños permanentes(AU)

Introduction: urinary tract infections are a frequent problem in the pediatric population. Recurrent urinary tract infection has a higher risk of kidney scars and glomerular damage. Objective: to describe frequency of hematuria, presence of proteins in urine, increase of blood pressure, and ultrasound findings in pediatric patients from 1 month to 14 years old with a diagnosis of urinary tract infection. Methods: cross-sectional descriptive study conducted from October 2014 to February 2016, in patients who attended the pediatric emergency´s service of the Hospital of San José with a diagnosis of urinary tract infection. Results: 125 patients with urinary tract infection were diagnosed, the median age was 2.75 years, it was more frequent in women (75.2 percent) and the majority of them received intrahospital treatment (58.4 percent). It was observed that in the recurrent urinary tract infection group the finding of hematuria was more frequent, than in the group of urinary tract infection as a first episode (21.9 vs. 11.8 percent), as well as the renal malformations (32,0 vs. 14.5percent). In blood pressure, there were no large differences among the groups, with a rate of high blood pressure frequency in hospitalized patients of 19.6 percent for first episode, and 18.2 percent in recurrent patients. The abnormal presence of proteins in urine was of 35.4 percent in the group of urinary tract infection first episode(AU) Conclusions: the presence of hematuria, renal malformations and increase of blood pressure in patients with urinary tract infection are frequent. There is an unusual frequency of patients with high blood pressure and proteins in the urine, probably due to false positives. It must be addressed the ambulatory monitoring of urinalysis and blood pressure to ensure that it normalizes or to detect permanent damage

Alteraciones urinarias en niños con primera infección urinaria e infección urinaria recurrente / Urinary disorders in children with first urinary infection and recurrent urinary tract infection

Introducción: las infecciones del tracto urinario son un problema frecuente en la población pediátrica. La infección del tracto urinario recurrente tiene mayor riesgo de cicatrices renales y daño glomerular. Objetivo: describir la frecuencia de hematuria, presencia de proteínas en orina, elevación en tensión arterial y hallazgos ecográficos en pacientes pediátricos desde 1 mes hasta 14 años de edad, con diagnóstico de infección del tracto urinario. Métodos: estudio descriptivo transversal, realizado entre octubre de 2014 y febrero de 2016, en pacientes que asistieron al servicio de urgencias pediátricas del Hospital de San José, con diagnóstico de infección del tracto urinario. Resultados: se diagnosticaron 125 pacientes con infección del tracto urinario, la mediana de edad fue de 2,75 años, más frecuente en mujeres (75,2 por ciento) y la mayoría recibió tratamiento intrahospitalario (58,4 por ciento). Se observó que en el grupo de infección del tracto urinario recurrente fue más frecuente el hallazgo de hematuria, que en la infección del tracto urinario primer episodio (21,9 vs. 11,8 por ciento), al igual que las malformaciones renales (32,0 vs. 14,5 %). En la tensión arterial no se observaron grandes diferencias entre los grupos, con una frecuencia de tensión arterial elevada en hospitalizados de 19,6 por ciento para primer episodio, y 18,2 por ciento en recurrentes. La presencia anormal de proteínas en orina fue de 35,4 por ciento en el grupo de infección del tracto urinario primer episodio. Conclusiones: son frecuentes la presencia de hematuria, malformaciones renales y elevación de la tensión arterial en pacientes con infección del tracto urinario. Se encuentra una frecuencia inusual de pacientes con elevación de la presión arterial y proteínas en orina, probablemente por falsos positivos. Se debe insistir en seguimiento ambulatorio del uroanálisis y la tensión arterial, para garantizar que esta se normalice o para detectar daños permanentes(AU)

Introduction: urinary tract infections are a frequent problem in the pediatric population. Recurrent urinary tract infection has a higher risk of kidney scars and glomerular damage. Objective: to describe frequency of hematuria, presence of proteins in urine, increase of blood pressure, and ultrasound findings in pediatric patients from 1 month to 14 years old with a diagnosis of urinary tract infection. Methods: cross-sectional descriptive study conducted from October 2014 to February 2016, in patients who attended the pediatric emergency´s service of the Hospital of San José with a diagnosis of urinary tract infection. Results: 125 patients with urinary tract infection were diagnosed, the median age was 2.75 years, it was more frequent in women (75.2 percent) and the majority of them received intrahospital treatment (58.4 percent). It was observed that in the recurrent urinary tract infection group the finding of hematuria was more frequent, than in the group of urinary tract infection as a first episode (21.9 vs. 11.8 percent), as well as the renal malformations (32,0 vs. 14.5percent). In blood pressure, there were no large differences among the groups, with a rate of high blood pressure frequency in hospitalized patients of 19.6 percent for first episode, and 18.2 percent in recurrent patients. The abnormal presence of proteins in urine was of 35.4 percent in the group of urinary tract infection first episode(AU) Conclusions: the presence of hematuria, renal malformations and increase of blood pressure in patients with urinary tract infection are frequent. There is an unusual frequency of patients with high blood pressure and proteins in the urine, probably due to false positives. It must be addressed the ambulatory monitoring of urinalysis and blood pressure to ensure that it normalizes or to detect permanent damage

Erratum: Erratum: Multiple Hereditary Exostoses: Report of an EXT2 Gene Mutation in a Colombian Family.

[This corrects the article DOI: 10.1055/s-0038-1636998.].

Detección y tipificación de virus del papiloma humano en biopsias de carcinoma ductal infiltrante y lesiones benignas de mama en mujeres venezolanas

Objetivo: Realizar la detección y tipificación del virus del papiloma humano (VPH) en muestras de biopsias de tejido mamario con carcinoma ductal infiltrante. Métodos: Estudio descriptivo de corte transversal de 57 biopsias de carcinoma ductal infiltrante, y 41 biopsias de lesiones benignas de mama de pacientes venezolanas, estas fueron evaluadas utilizando la técnica PCR-RFLP en busca de la presencia del genoma del virus de papiloma humano. El riesgo OR fue evaluado mediante análisis estadístico con el paquete SPSS 12.0. Resultados: Treinta y tres (57,9 %) de las muestras de carcinoma ductal infiltrante tuvieron un resultado positivo para virus de papiloma humano, 19 de ellas pudieron ser tipificadas como: VPH-6b 15,15 %; VPH-11 3,03 %; VPH-18 12,12 %; VPH-33 27,27 %; VPH-45 3,03 % y VPH-58 3,03 %; de este grupo el 42,4 % fueron positivas no determinadas para la presencia de ADN del virus. Seis biopsias de lesiones benignas (14,6 %), presentaron infección por virus de papiloma humano, determinándose para ellas los tipos VPH-6b 33,33 %, VPH-11 16,67%, VPH-33 16,67% y 33,33 % positivas no determinadas. Se determinó estadísticamente que la presencia de virus de papiloma humano en tejido mamario aumenta 10,77 veces la posibilidad de desarrollar carcinoma ductal infiltrante. Conclusiones: Los hallazgos corroboran los resultados de otros investigadores, colocando al virus de papiloma humano como posible agente involucrado en la inmortalización de las células epiteliales de la mama.

Objective: To perform the detection and typing of human papilloma (HPV) virus in biopsy samples of breast tissue invasive ductal cancer. Methods: Cross-sectional study of 57 biopsies of invasive ductal carcinoma, and 41 biopsies of benign breast lesions of Venezuelan patients were evaluated using the PCR-RFLP technique for the presence of the human papillomavirus genome. The OR risk was evaluated by statistical analysis using SPSS package. Results: Thirty-three (57.9%) of invasive ductal carcinoma samples had a positive result for human papillomavirus, 19 of them could be classified as: HPV-6b 15.15%; HPV-11 3.03%; HPV-18 12.12%; HPV-33 27.27%; HPV-45 3.03% and HPV-58 3.03%. This group 42.4% were positive not determined for the presence of virus DNA. Six biopsies of benign lesions (14.6%) had human papillomavirus infection, determining for themselves the types HPV-6b 33.33%, 16.67% HPV-11, HPV-33 16.67% and 33.33% not determined positive. It is statistically determined that the presence of human papillomavirus in breast tissue 10.77 times increases the possibility of developing invasive ductal carcinoma. Conclusions: These findings corroborate the results of other researchers, placing human papillomavirus as a possible agent involved in the immortalization of epithelial cells of the breast.

Estandarización de un sistema PCR multiplex para la determinación molecular de infertilidad masculina idiopática

Objetivo: Diseñar y optimizar sistemas de reacción en cadena de polimerasa individuales y multiplex para la detección de microdeleciones de genes asociados a infertilidad masculina en el cromosoma Y. Métodos: Se estandarizaron sistemas de reacción en cadena de polimerasa multiplex utilizando oligonucleótidos STS (Sequence Target Site) específicos asociados a infertilidad masculina, con previa estandarización de cada par de oligos en reacciones individuales. Resultados: Se logró estandarizar 7 sistemas individuales y 2 sistemas multiplex de alta sensibilidad y especificidad que pueden indicar la presencia o ausencia de un gen, en este caso, son utilizados para indicar la mutación por microdeleción de algún fragmento específico de Yq que conlleva a la inactivación de un gen. Conclusiones: Se pudo realizar la estandarización de dos sistemas multiplex para el análisis de microdeleciones del cromosoma Y asociados a infertilidad masculina como una herramienta molecular para el diagnóstico rápido y preciso de esta patología. El amplificado del marcador RBM1 no pudo ser incluido en ninguna de los dos multiplex estandarizados, no obstante, se sugiere el estudio de otros marcadores de infertilidad masculina que puedan ser incluidos en la estandarización de nuevos multiplex.

Objective: To design and optimize individual systems and multiplex polymerase chain reaction for detection of microdeletions of male infertility associated genes on the Y chromosome. Methods: multiplex polymerase chain reaction systems were standardized using oligonucleotides STS (Sequence Target Site) specific to male infertility associated with prior standardization of each pair of oligos in individual reactions. Results: It was possible to standardize 7 individual systems and two multiplex systems high sensitivity and specificity that may indicate the presence or absence of a gene, in this case, are used to indicate the mutation microdeletion of a specific fragment Yq leading to the inactivation of a gene. Conclusions: standardization could make two multiplex systems for the analysis of microdeletions of the Y chromosome associated with male infertility as a molecular tool for rapid and accurate diagnosis of this disease. The amplified RBM1 marker could not be included in either standard multiplex, despite the studies of other markers of male infertility is suggested to be included in new in the standardization of new multiplexes.

Detección molecular de VPH por PCR-RFLP en muestras del área ano-genital de hombres en el Estado Mérida, Venezuela

Objetivo: Determinar el porcentaje de positividad y tipo de virus de papiloma humano en el área genitoanal de hombres, relación con la edad, sitio anatómico y tipo de muestra, en el Departamento de Biología, Laboratorio de Biología y Medicina Experimental Facultad de Ciencias. LABIOMEX Universidad de Los Andes. Métodos: Estudio descriptivo, transversal, de 882 muestras del área genito-anal de hombres que acudieron voluntariamente a la consulta para detección y tipificación de virus de papiloma humano mediante reacción en cadena de la polimerasa y análisis de restricción. Resultados: El porcentaje de positividad y tipo de virus del papiloma humano encontrado fue de 49,9.% donde el virus de papiloma humano de alto riesgo fue de 46,2 %, (VPH16: 12,4 %, VPH18: 11,0 % y VPH31: 6 %) y virus de papiloma humano de bajo riesgo el 53,8 % (VPH6: 35,8 % y VPH11: 12 %). El cepillado fue más eficiente en rendir ADN de buena calidad. El rango de edad 20-26 años fue el más numeroso. Se encontró una fuerte asociación entre muestras de la región anal y virus de papiloma humano de alto riesgo comparado con las muestras de otras áreas genitales (Chi-cuadrado=7,405, P=0,05). Conclusiones: El porcentaje de positividad de virus de papiloma humano encontrado en este estudio es alto, con una frecuencia importante de virus de papiloma humano de alto riesgo, y con una fuerte asociación con muestras de la región anal. La tasa de cáncer cervical en nuestro país es alta, en comparación con otros países de la región, la alta frecuencia de virus de papiloma humano de alto riesgo en nuestra población masculina puede ser un factor que contribuya a que más mujeres se infecten con tipos oncogénicos y desarrollen esta enfermedad.

Objective: Determine the percentage of positivity and human papillomavirus type in the genital and anal area in men, related to age, anatomical site and type of sample. Method: A descriptive, cross-sample of 882 genito-anal area of men who came voluntarily to the clinic for human papillomavirus detection and typing by PCR-RFLP. Environment: Faculty of Sciences, Laboratory of Experimental Biology and Medicine LABIOMEX, Universidad de Los Andes. Merida, Merida, Venezuela. Results: The human papillomavirus percentage of positivity was 49.9 % where human papillomavirus high risk was 46.2 % (HPV16: 12.4 %, HPV18: 11.0 % and HPV31: 6 %) and human papillomavirus low risk was 53.8 % (HPV6: 35.8 % and VPH11: 12 %). Brushing was more efficient in yield good quality DNA. The age range 20-26 years was the largest. We found a strong association between samples of the anal region and human papillomavirus high risk compared with samples from other genital areas (Chi-square = 7.405, P = 0.05). Conclusions: The human papillomavirus prevalence found in this study was high, with a high frequency of human papillomavirus high risk and with a strong association with samples of the anal region. Cervical cancer rates in our country is high compared with other countries in the region, the high frequency of human papillomavirus high risk in our male population may be a contributing factor to more women becoming infected with oncogenic types and develop this disease.

[Characterization of extended-spectrum ß-lactamases-producing Klebsiella pneumoniae isolates of two intensive care units]. / Caracterización de cepas de Klebsiella pneumoniae productora de ß-lactamasa de espectro extendido aisladas en dos unidades de cuidados intensivos.

BACKGROUND: Extended-spectrum ß-lactamase (ESBL)- producing Klebsiella pneumonia is more the frequently implied bacterium infections nosocomiales in the Neonatal High Risk Unit (NHRU), and adult intensive care unit (aICU) in the The Andes University Hospital Autonomy Institute. AIM: To determine the microbiological and molecular characteristics associated with infections caused by this bacterium. MATERIAL AND METHODS: 17 strains of K. pneumoniae isolated from neonates from NHRU with nosocomial infections and 11 isolates from patients hospitalized in the aICU between June to November 2009 were characterized by in vitro agar diffusion test and MIC, the production of ß-lactamase, the presence of blaTEM blaSHV y blaCTX-M genes and genotyping. RESULTS: A high percentage of resistance to ß-lactams and the presence of ESßL TEM, SHV and CTX-M in 41.2 y 82% of strains obtained at HRNU and at aICU respectively were detected. BLEE (+) phenotype was detected in 17,6% of HRNU strains and 91% of aICU strains. Genotypic analysis by REP-PCR detected several clones suggesting that resistant clones of ESBL-producing K. pneumoniae are endemic not only specific circulation and transmission of a single common among patients. CONCLUSION: K. pneumoniae strains that produce ß-lactamases circulate in these critical units studied representing a therapeutic challenge that is necessary to fight against.

Caracterización de cepas de Klebsiella pneumoniae productora de β-lactamasa de espectro extendido aisladas en dos unidades de cuidados intensivos / Characterization of extended-spectrum β-lactamases-producing Klebsiella pneumoniae isolates of two intensive care units

Background: Extended-spectrum β-lactamase (ESBL)- producing Klebsiella pneumonia is more the frequently implied bacterium infections nosocomiales in the Neonatal High Risk Unit (NHRU), and adult intensive care unit (aICU) in the The Andes University Hospital Autonomy Institute. Aim: To determine the microbiological and molecular characteristics associated with infections caused by this bacterium. Material and Methods: 17 strains of K. pneumoniae isolated from neonates from NHRU with nosocomial infections and 11 isolates from patients hospitalized in the aICU between June to November 2009 were characterized by in vitro agar diffusion test and MIC, the production of β-lactamase, the presence of blaTEM blaSHV y blaCTX-M genes and genotyping. Results: A high percentage of resistance to β-lactams and the presence of ESβL TEM, SHV and CTX-M in 41.2 y 82% of strains obtained at HRNU and at aICU respectively were detected. BLEE (+) phenotype was detected in 17,6% of HRNU strains and 91% of aICU strains. Genotypic analysis by REP-PCR detected several clones suggesting that resistant clones of ESBL-producing K. pneumoniae are endemic not only specific circulation and transmission of a single common among patients. Conclusion: K. pneumoniae strains that produce β-lactamases circulate in these critical units studied representing a therapeutic challenge that is necessary to fight against.

Introducción: Klebsiella pneumoniae productora de β-lactamasa de espectro extendido (βLEE) es la bacteria más frecuentemente implicada en infecciones nosocomiales en las Unidades de Alto Riesgo Neonatal (UARN) y Cuidados intensivos de adulto (UCIa) del Instituto Autónomo Hospital Universitario de Los Andes. Objetivo: Determinar las características microbiológicas y moleculares asociadas a las infecciones causadas por esta bacteria. MaterialyMétodos: Se caracterizaron la susceptibilidad in vitro por difusión en agar y CIM, la presencia de β-lactamasa y la presencia de genes blaTEM blaSHV y blaCTX-M y la genotipificación en 17 cepas de K. pneumoniae aisladas de neonatos con infección nosocomial, así como 11 aisladas de pacientes hospitalizados en la UCIa entre junio y noviembre de 2009. Resultados: Esta investigación reveló un alto porcentaje de resistencia a β-lactámicos y la presencia de genes tipo blaTEM, blaSHV y blaCTX-M en 41,2 y 82% de las cepas procedentes de UARN y UCIa, respectivamente. Producción de BLEE se detectó en 17,6% cepa de UARN y en 91% de las pertenecientes a UCIa. Mediante el análisis genotípico REP-PCR se detectaron varias clones lo cual sugiere que existen clones resistentes de K. pneumoniae productora de βLEE específicas no endémicas más que la circulación y transmisión de un aislado habitual entre pacientes. Conclusión: En las unidades críticas estudiadas circulan cepas de K. pneumoniae productoras de β-lactamasas constituyendo un problema terapéutico importante y necesario de combatir.

Diseño de un sistema de PCR para la detección del virus del papiloma humano mediante el uso de oligonucleótidos degenerados de la región E6 / Desing of PCR system for detection of human papilloma virus by using degenerate oligonucleotides of the E6 region

Diseñar y estandarizar un sistema de PCR para detectar un amplio espectro de tipos de VPH en una sola reacción utilizando como blanco la región E6 del genoma viral. Utilizando secuencias del gen E6 de distintos tipos de VPH del NCBI y las herramientas informáticas Mult Alin versión 5.4.1. y Oligo Analyzer 1.1.2 se diseñaron oligonucleótidos degenerados que permitían la amplificación de un fragmento de aproximadamente 214 pb. Las muestras seleccionadas incluyen: 25 muestras de ADN, cada una positiva para un solo tipo de VPH tipificados por el sistema PCR-RFLP MY11/MY09 del gen L1. Se obtuvieron amplificados de aproximadamente 214 pb de 25 tipos distintos de VPH, se detectaron amplificados de E6 en muestras negativas para el sistema MY09/MY11. Se pudo determinar que el diseño de los oligonucleótidos degenerados fue altamente eficiente para la amplificación de por lo menos 25 tipos distintos de VPH lo que facilitaría la detección en muestras clínicas

Design and standardize a PCR system to detect a broad spectrum of HPV types in a single reaction using target the E6 region of the viral genome. Using E6 gene sequences of different HPV types and NCBI tools Mult Alin Oligo Analyzer version 1.1.2 5.4.1. Degenerate oligonucleotides were designed that allowed amplification of a fragment of approximately 214 bp. Selected samples include: 25 DNA samples, each positive for a single HPV type typified by PCR-RFLP system MY11/MY09 L1 gene. We obtained approximately 214 bp amplified from 25 different HPV types were detected in samples amplified from E6 negative MY09/MY11 system. It was determined that the design of degenerate oligonucleotides was highly efficient for amplification of at least 25 different HPV types which would facilitate detection in clinical samples

Identificación de la mutación R579X en el exón 18 del gen RB1, en pacientes con lesiones cervicales asociadas a infección por virus del papiloma humano / Identification of the R579X mutation in exon 18 of the RB1 gene in patients with cervical infection associated with human papilloma virus

Identificar la mutación R579X ubicada en el exón 18 gen RB1, en pacientes con lesiones cervicales asociadas a infección por virus de papiloma humano mediante PCR-SSCP, PCR-RFLP y secuenciamiento. Estudio de 72 muestras de hisopado/cepillado de cuello uterino provenientes de 36 mujeres sanas, y 36 con infección por virus de papiloma humano y con presencia de lesiones cervicales a las cuales se les realizó la extracción de ADN, la amplificación del exón 18 del gen RB1 mediante PCR, y el análisis mediante PCR-SSCP, Secuenciamiento y PCR-RFLP. El análisis por PCR-SSCP reveló dos patrones diferentes de corrida y mediante el secuenciamiento se logró identificar la mutación R579X en el exón 18 del gen supresor de tumores RB1 en el 30,56 por ciento de las muestras experimentales. La mutación R579X encontrada en las muestras experimentales conlleva a la formación de una proteína truncada no funcional, y aunado a esto, el virus de papiloma humano podría estar favoreciendo a la inmortalización de las células que presentan dicha mutación

To identify the mutation R579X in exon 18 located RB1 gene in patients with cervical lesions associated with human papilloma virus infection by PCR-SSCP, PCR-RFLP and sequencing. Study of 72 samples of cervical brushing of the healthy women, infected with human papilloma virus and with cervical lesions that required DNA extraction, PCR amplification of exon 18 of the RB1 gene, PCR-SSCP, Sequencing and PCR-RFLP analysis. PCR-SSCP analysis revealed two different band patterns. We indentified the R579X mutation in exon 18 of the RB1 tumor suppressor gene in 30.56 percent of the experimental samples. The mutation found in the experimental samples leads to the formation of a non-functional truncated protein, in adition, human papilloma virus infection might contributed to the immortalization of cells with this mutation