[Detection of human papillomavirus in gingival fluid of Venezuelan HIV patients with periodontal disease]. / Detección de virus papiloma humano en el fluido gingival de pacientes con inmunodeficiencia humana y enfermedad periodontal.

Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the IIPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 +/- 324,244 copy/mL) than in the HPV-patients (10,246 +/- 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with IIPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

Detección de Virus Papiloma Humano en el fluido gingival de pacientes con inmunodeficiencia humana y enfermedad periodontal / Detection of Human Papillomavirus in gingival fluid of Venezuelan HIV patients with periodontal disease

Evidencias sugieren que los virus pueden participar en la activación de la enfermedad periodontal, permitiendo el sobrecrecimiento de bacterias periodontopatógenas. El objetivo del estudio fué la detección molecular de VPH en fluido gingival (FG) de pacientes VIH+ con enfermedad periodontal. Se evaluaron muestras de FG de 20 pacientes VIH+ con enfermedad periodontal que asistieron al Centro de Atención de Pacientes con Enfermedades Infecciosas (CAPEI) de la Facultad de Odontología de la Universidad Central de Venezuela, 13 bajo terapia antirretroviral (HAART) y 7 VIH+ sin HAART. Se incluyeron 7 pacientes seronegativos con periodontitis crónica y como grupo control 7 pacientes seronegativos periodontalmente sanos. Se extrajo el ADN, se amplificó la región L1 de VPH con primers MY09 y MY11. Las muestras VPH+ fueron genotipificadas para los tipos 6, 11, 16, 18, 31 y 45. VPH fue detectado en 46% de los pacientes VIH+ bajo terapia. El contaje CD4+ en la población VPH+ no presentó diferencias con el grupo VPH-, y la carga viral mostró valores promedio significativamente mayores (200.470± 324.244 copias/mL) con respecto a los pacientes VPH- (10.246±23.805 copias/mL). Las muestras VPH+ presentaron los genotipos 6 y 11, de los cuales 66,6% estaban coinfectados con ambos tipos. Las condiciones periodontales no presentaron diferencias entre los individuos con doble infección viral por VPH y VIH, y los que solo portaban VIH. VPH fue detectado solamente en fluido gingival de pacientes VIH+ con HAART, indicando que esta terapia puede influir en el estado inmunológico independientemente de las condiciones periodontales.

Evidence suggests that viruses may be involved in the activation of periodontal disease, allowing the overgrowth of periodontal pathogens. The purpose of the present study was to detect the presence of Human Papillomavirus (HPV) in gingival crevicular fluid (GCF) in HIV+ Venezuelan patients with periodontal disease. We evaluated GCF samples from 20 HIV+ patients with periodontal disease from the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to establish their periodontal conditions, 13 under HAART (antiretroviral therapy) and 7 without HAART. Seven seronegative patients with chronic periodontitis and 7 seronegative patients, without periodontal disease were included. DNA extraction was performed, the consensus primers MY09 and MY11 for the HPV L1 region were used for PCR amplification. Genotipification was made for the 6, 11, 16, 18, 31 and 45 genotypes. HPV were detected in 46% of HIV+ patients under therapy. The CD4 cell counts in the HPV+ patients were not significantly different from the HPV-group. The viral load in the HPV+ group was significantly higher (200,470 ± 324,244 copy/mL) than in the HPV- patients (10,246 ± 23,805 copy/mL). Genotypes 6 and 11 were observed in the HPV positive samples, of which 4/6 (66.6%) presented coinfection with both types. No significant differences in the periodontal conditions were observed between patients with HPV-HIV infection related to patients with only HIV. HPV was detected only in the gingival crevicular fluid of HIV+ patients under HAART independently of the periodontal conditions.

Detección de microbacterias del complejo tuberculosis en tejido periodontal: aplicado biología molecular / Detection of mycobacteria tuberculosis complex in periodontal tissue: molecular biology applied

El objetivo de esta investigación fue la detección molecular de micobacterias del complejo tuberculosis en el tejido periodontal afectado de un grupo de pacientes VIH+ y VIH- con tuberculosis pulmonar. Se incluyeron 20 pacientes, 10 VIH+ y 10 VIH- con diagnóstico microbiológico convencional de tuberculosis, provenientes de la consulta de Tisiología, y de Sala de VIH, del Hospital Simón Bolívar, Caracas. De cada uno de los pacientes fue obtenido el consentimiento informado previo al inicio de la investigación, se le realizó una minuciosa historia clínica y un examen clínico bucal, con el fin de establecer la condición periodontal de cada uno de los sujetos participantes en este estudio. Se tomaron muestras de tejido periodontal afectado de cada uno de los pacientes con indicación de cirugía periodontal. Resultados: 50% (10/20) de los pacientes con TBC VIH- presentaron gingivitis crónica localizada y el 30% (3/20) de los pacientes con TBC VIH+ presentaron gingivitis crónica generalizada. El grupo restante mostró gingivitis localizada y cuadros sugerentes de periodontitis. Por medio de la reacción en cadena de la polimerasa (PCR) en 60% (12/20) de las muestras de tejido periodontal se detectó la secuencia IS6110, que está presente en micobacterias del complejo tuberculosis. La amplificación de blancos moleculares es una metodología sensible para la detección de este grupo de microorganismos en enfermedad periodontal y pudiera ser utilizada en el diagnóstico diferencial de lesión de la cavidad bucal

Human papillomavirus and Epstein Barr virus in oral hairy leukoplakia among HIV positive Venezuelan patients.

Oral hairy leukoplakia (OHL) is commonly found in individuals infected with HIV and represents the most frequent oral manifestation. The purpose of this study was to detect the presence of Human Papillomavirus (HPV) and Epstein Barr Virus (EBV) in OHL of HIV+ Venezuelan patients. We evaluated 21 HIV+ adult patients with clinically present OHL lesions: 11 under antiretroviral therapy, 10 without therapy, and 10 oral mucosal samples as controls. Nested-PCR was used to detect EBV and HPV infection. The INNO-LiPA HPV Genotyping v2 was applied to determine the HPV genotype. The EBV genome was found in 16/21 (76%) of the HIV+ patients with OHL. No difference was observed in EBV+ and EBV- patients related to antiretroviral therapy viral load and CD4+ Tcell coant. HPV-DNA was observed in 7/21 HIV positive cases (33%). The HPV genotypes detected were: 6, 11, 31, 33, 52, and 56/74. The most frequently HPV found was genotype 6 in 7/7, while two cases were HPV-11 and two HPV-52. Of the positive cases, 5/7 (71%) presented co-infection with more than one HPV genotype and 4/7 (57%) had HPV coinfection with high and low risk types. No case was EBV or HPV positive in the control group. In this study, a higher EBV prevalence was observed in OHL-HIV+ patients, confirming the etiologic role in this entity. A considerable number of cases were positive for HPV infection, and many patients presented coinfection with more than one HPV genotype as well as the presence of high oncogenic risk HPV in OHL.

Human papillomavirus and Epstein Barr virus in oral hairy leukoplakia among HIV positive Venezuelan patients

Oral hairy leukoplakia (OHL) is commonly found in individuals infected with HIV, and represents the most frequent oral manifestation. The purpose of this study was to detect the presence of Human Papillomavirus (HPV) and Epstein Barr Virus (EBV) in OHL of HIV+ Venezuelan patients. We evaluated 21 HIV+ adult patients with clinically present OHL lesions: 11 under antiretroviral therapy, 10 without therapy, and 10 oral mucosal samples as controls. Nested-PCR was used to detect EBV and HPV infection. The INNO-LiPA HPV Genotyping v2 was applied to determine the HPV genotype. The EBV genome was found in 16/21 (76%) of the HIV+ patients with OHL. No difference was observed in EBV+ and EBV- patients related to antiretroviral therapy viral load and CD4+ T cell count. HPV-DNA was observed in 7/21 HIV positive cases (33%). The HPV genotypes detected were: 6, 11, 31, 33, 52, and 56/74. The most frequently HPV found was genotype 6 in 7/7, while two cases were HPV-11 and two HPV-52. Of the positive cases, 5/7 (71%) presented co-infection with more than one HPV genotype and 4/7 (57%) had HPV coinfection with high and low risk types. No case was EBV or HPV positive in the control group. In this study, a higher EBV prevalence was observed in OHL-HIV+ patients, confirming the etiologic role in this entity. A considerable number of cases were positive for HPV infection, and many patients presented coinfection with more than one HPV genotype as well as the presence of high oncogenic risk HPV in OHL.

El proposito del presente estudio fue detectar la presencia de virus papiloma humano (VPH) y Epstein Barr (VEB) en Leucoplasia Vellosa Oral (LVO) de pacientes VIH positivos. Se evaluaron 21 pacientes adultos VIH positivos con lesiones clinicas presentes de LVO y 10 casos controles de mucosa sana. Para el diagnostico molecular de VPH y EBV se utilizo Nested PCR. La determinacion de los genotipos se realizo mediante el kit HPV INNO-LiPA genotyping v2. La presencia de genoma de VEB se demostro en un alto porcentaje (76%) en 16/21 de los pacientes VIH positivos con LVO. No se observo relacion entre los pacientes VEB+ y VEBcon el uso de terapia antirretroviral, la carga viral y el contaje de celulas T CD4+. Se demostro la presencia de ADN-VPH en 7/21 (8%) de los casos VIH positivos. Los genotipos de VPH detectados fueron 6, 11, 31, 33, 52, 56/74. El genotipo 6 fue el mas frecuentemente observado en 7/7, dos casos fueron VPH 11 y dos VPH 52. De los casos positivos 5/7 (71%) presentaron coinfeccion con mas de un genotipo de VPH y en 4/7 (57%) se evidencio coinfeccion con tipos de alto y bajo riesgo oncogenico. En el presente estudio se observo una alta prevalencia de VEB en pacientes VIH positivos con LVO, confirmando el papel etiologico en esta entidad. Un considerable numero de casos fueron positivos para VPH. Se observo la presencia de coinfeccion con mas de un tipo viral, asi como la presencia de VPH de alto riesgo.

Human papillomavirus in ameloblastoma.

OBJECTIVE: Ameloblastomas are benign epithelial tumors of odontogenic origin, with a high recurrence rate and local aggressiveness. A few preliminary studies have demonstrated HPV presence mainly in peripheral ameloblastomas. The purpose of this study was to detect HPV-DNA in intraosseous ameloblastomas. METHODS: Eighteen cases of intraosseous ameloblastomas of different histological variants were selected. Immunohistochemistry, CISH, nested-PCR, and INNOLiPA HPV Genotyping v2 were used. RESULTS: The predominant age group was between the third and fourth decades of life. Males were more affected with 61% and females represented 39%. Of the 18 cases, 7 were solid multicystic tumors and 11 were unicystic. Of the histological variants, the plexiform represented 3 (17%) of the 18, 2 (11%) were follicular, 2 (11%) were acanthomatous, and 1 (6%) was desmoplastic. All cases were HPV negative by immunohistochemistry and CISH. HPV-DNA was detected in 6 (33%) of the cases by nested-PCR. HPV 6 was the most frequent genotype in 4 (66%) of the 6. Two cases presented a mixture of HPV 16, 33, and HPV 6, 42 respectively. Four of the unicystic ameloblastomas were HPV positive; of these, all presented koilocytic changes and were associated with dentigerous cysts, whereas only 2 positive cases corresponded to solid ameloblastomas. None of the positive cases were related to recurrence rate. CONCLUSIONS: We may conclude that HPV low and high risk was detected in our sample of intraosseous ameloblastomas. HPV positivity was observed more in the unicystic cases than solid types.

Helicobacter pylori cagA and vacA genotypes in Cuban and Venezuelan populations.

The aim of this study was to determine the presence of Helicobacter pylori cytotoxin-associated gene (cagA)/vacuolating cytotoxin gene (vacA) among patients with chronic gastritis in Cuba and Venezuela. Gastric antrum biopsies were taken for culture, DNA extraction and PCR analysis. Amplification of vacA and cagA segments was performed using two regions of cagA: 349 bp were amplified with the F1/B1 primers and the remaining 335 bp were amplified with the B7629/B7628 primers. The VA1-F/VA1-R set of primers was used to amplify the 259-bp (s1) or 286-bp (s2) product and the VAG-R/VAG-F set of primers was used to amplify the 567-bp (m1) or 642-bp (m2) regions of vacA. cagA was detected in 87% of the antral samples from Cuban patients and 80.3% of those from Venezuelan patients. All possible combinations of vacA regions were found, with the exception of s2/m1. The predominant combination found in both countries was s1/m1. The percentage of cagA+ strains was increased by the use of a second set of primers and a greater number of strains was amplified with the B7629/B7628 primers in the Cuban patients (p = 0.0001). There was no significant difference between the presence of the allelic variants of vacA and cagA in both populations. The predominant genotype was cagA+/s1m1 in both countries. The results support the necessary investigation of isolates circulating among the human population in each region.

Helicobacter pylori cagA and vacA genotypes in Cuban and Venezuelan populations

The aim of this study was to determine the presence of Helicobacter pylori cytotoxin-associated gene (cagA)/vacuolating cytotoxin gene (vacA) among patients with chronic gastritis in Cuba and Venezuela. Gastric antrum biopsies were taken for culture, DNA extraction and PCR analysis. Amplification of vacA and cagA segments was performed using two regions of cagA: 349 bp were amplified with the F1/B1 primers and the remaining 335 bp were amplified with the B7629/B7628 primers. The VA1-F/VA1-R set of primers was used to amplify the 259-bp (s1) or 286-bp (s2) product and the VAG-R/VAG-F set of primers was used to amplify the 567-bp (m1) or 642-bp (m2) regions of vacA. cagA was detected in 87 percent of the antral samples from Cuban patients and 80.3 percent of those from Venezuelan patients. All possible combinations of vacA regions were found, with the exception of s2/m1. The predominant combination found in both countries was s1/m1. The percentage of cagA+ strains was increased by the use of a second set of primers and a greater number of strains was amplified with the B7629/B7628 primers in the Cuban patients (p = 0.0001). There was no significant difference between the presence of the allelic variants of vacA and cagA in both populations. The predominant genotype was cagA+/s1m1 in both countries. The results support the necessary investigation of isolates circulating among the human population in each region.

Epstein Barr Virus detection and latent membrane protein 1 in oral hairyleukoplakia in HIV + Venezuelan patients

Purpose: To determine the prevalence of Epstein Barr Virus (EBV) in oral hairy leukoplakia lesions (OHL) inHIV+ Venezuelan patients. Material and Method: In this case study, we evaluated 21 HIV+ adult patients withclinically present OHL lesions, 11 who were undergoing antiretroviral therapy, 10 who were not undergoingtherapy and 10 HIV-negative adult patients with hyperkeratotic oral mucosal lesions. All of the subjects were assessedat the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinicallyexamined to detect oral mucosal lesions with the confirmed histopathologic diagnosis. Nested-PCR was used todetermine the EBV infection and the latent membrane protein-1 (LMP-1) expression by immunohistochemistry.Results: Of the subjects, 16/21 (76%) of the HIV+/AIDS patients tested positive for EBV, whereas 5/10 (50%) ofthe HIV-negative subjects tested positive for EBV. Conclusions: In the present study, a higher EBV prevalence wasobserved in HIV-positive patients when compared to HIV-negative patients without oral hairy leukoplakia, confirmingthe etiologic role in this entity. The LMP-1 in OHL patients who were both HIV+ and EBV+ was highlyexpressed (60%) at the epithelial basal cells. No association between the alcohol and tobacco consumption wasobserved among the EBV-positive cases (AU)

Epstein Barr Virus detection and latent membrane protein 1 in oral hairy leukoplakia in HIV+ Venezuelan patients.

PURPOSE: To determine the prevalence of Epstein Barr virus (EBV) in oral hairy leukoplakia lesions (OHL) in HIV+ Venezuelan patients. MATERIAL AND METHOD: In this case study, we evaluated 21 HIV+ adult patients with clinically present OHL lesions, 11 who were undergoing antiretroviral therapy, 10 who were not undergoing therapy and 10 HIV-negative adult patients with hyperkeratotic oral mucosal lesions. All of the subjects were assessed at the Infectious Disease Center, Faculty of Dentistry, Central University of Venezuela, and were clinically examined to detect oral mucosal lesions with the confirmed histopathologic diagnosis. Nested-PCR was used to determine the EBV infection and the latent membrane protein-1 (LMP-1) expression by immunohistochemistry. RESULTS: Of the subjects, 16/21 (76%) of the HIV+/AIDS patients tested positive for EBV, whereas 5/10 (50%) of the HIV-negative subjects tested positive for EBV. CONCLUSIONS: In the present study, a higher EBV prevalence was observed in HIV-positive patients when compared to HIV-negative patients without oral hairy leukoplakia, confirming the etiologic role in this entity. The LMP-1 in OHL patients who were both HIV+ and EBV+ was highly expressed (60%) at the epithelial basal cells. No association between the alcohol and tobacco consumption was observed among the EBV-positive cases.

Detección de Candida Albicans en lesiones de leucoplasia vellosa bucal de un grupo de pacientes venezolanos VIH+ / Detection of candida albicans in lesions of oral hairy leukoplakia a group of HIV + patients in Venezuela

El objetivo del presente estudio fue determinar la prevalencia de la infección por Candida albicans en lesiones de Leucoplasia Vellosa Bucal (LVB) en un grupo de pacientes VIH+ en una muestra de la población venezolana. Para ello se evaluaron 21 pacientes adultos VIH+, con lesiones clínicas de Leucoplasia Vellosa Bucal, 11 con terapia antiretroviral y 10 sin terapia, y 10 pacientes adultos VIH- con lesiones hiperqueratósicas de la mucosa oral provenientes del Centro para la Atención de Pacientes con Enfermedades Infectocontagiosas de la Facultad de Odontología de la Universidad Central de Venezuela. Los pacientes fueron examinados clinícamente para la detección de lesiones presentes en la mucosa oral, confirmándose el diagnóstico con el estudio histopatológico. La infección por C albicans se determinó mediante biopsias de las lesiones para su estudio histopatológico empleando dos técnicas diferentes, coloración de Pass y método de Grocott, y cultivo microbiológico en medio Agar Sabouraud Resultados: En pacientes VIH+ según los distintos métodos empleados se pudo observar que mediante el cultivo en Agar Sabouraud 6/21 (29%) de los pacientes fueron positivos y 15/21 (71%) fueron negativos. Con respecto a la coloración de Grocott, todos los 21 pacientes fueron positivos, mientras que con la coloración de PAS, 17/21 (81%) de los pacientes fueron positivos y 4/21 (19%) fueron negativos. En el caso de los pacientes VIH- con leucoplasia bucal, 1/10 (10%) resultó positivo mediante el cultivo en Agar Sabouraud y 9/10 (90%) fueron negativos. Mediante la coloración de Grocott todos los pacientes (10) fueron negativos para la presencia del hongo, e iguales resultados fueron obtenidos con la coloración de PAS. Conclusión: Se observó una mayor prevalencia de la infección por C. albicans en pacientes VIH+ lo que demuestra que la presencia del hongo, constituye un factor fuertemente asociado a los pacientes con LVB

The aim of this study was to determine the prevalence of Candida albicans infection in Oral Hairy Leukoplakia (OHL) in Venezuelan HIV+ population. We evaluated 21 HIV+ adults patients with the clinical diagnosis of OHL, 11 under antiretroviral therapy, 10 without and 10 HIV negative adults with hyperkeratotic lesions on the oral mucosa as a control group, from the Center of Infectious diseases, Faculty of Dentistry, Central University of Venezuela. All patients were clinically examined and definitive histopathological diagnosis was made. C albicans was detected by two different histochemical methods: PAS and Groccot and microbiological culture using Agar Sabouraud medium. The results demonstrated that 6/21 (29%) patients were positive by microbiological culture and 15/21 (71%) were negative. All the patients were positive by Grocott special stain while 17/21 (81%) were positive using PAS. In the control group, 1 patient was positive by culture and 9/10 (90%) were negative. Additionally, all the patients were negative by Grocott and PAS. In conclusion, we observed a higher prevalence of C. albicans infection in HIV+ patients, suggesting that the presence of this microorganism may constitute an associated factor in OHL

Fluconazol: Seguridad y Eficacia en el Control de la Estomatitis Sub-Protésica Asociada a Candida

La Estomatitis Sub-Protésica (E.S.P.) describe cambios patógenos encontrados en los tejidos de soporte de la dentadura. Dichos cambios se caracterizan por la presencia de un eritema, debido entre otros factores a la proliferación de microorganismos del Género Candida. Es por ello que el objetivo principal del presente trabajo fue comprobar la eficacia del Fluconazol (Diflucán®) tanto por vía oral como en forma tópica sobre la mucosa afectada, en un grupo de pacientes con diagnóstico de E.S.P. inducida por Candida. En esta investigación se seleccionaron 30 pacientes que acudieron al Servicio de Clínica Estomatológica "Magdalena Mata de Henning" con diagnóstico presuntivo de E.S.P., éstos fueron divididos en 3 grupos de 10 pacientes cada uno: Grupo A: Los cuales fueron medicados una vez al día con Fluconazol (Diflucán®), administrado por vía oral, y aplicado tópicamente en forma de suspensión en la prótesis, en ambos casos durante 14 días; Grupo B: Quienes fueron medicados una vez al día con Fluconazol (Diflucán®), administrado por vía oral por 14 días, pero sin medicación para la prótesis. Grupo C: A quienes se les administró un placebo una vez al día, durante 14 días pero sin aplicación de producto sobre la prótesis. De igual manera, se les dictaron instrucciones a todos los pacientes en relación con el mantenimiento de una adecuada higiene bucal, así como realizarla previa administración del tratamiento antimicótico. En este estudio se demostró que el tratamiento antimicótico con Diflucán®, es eficaz contra la E.S.P. inducida por Candida.

Denture Stomatitis (DS), defined as a confluent erythema and oedema of the denture bearing area of the palate. The ocurrence of Candida can be demonstrated in the mouth in persons with DS to a significantly higher degree than in healthy controls, and differents antifungal agents has frecuently been used in dental practice in order to cure the disease. The purpose of this study was to evaluate the clinical efficacy and the safety of oral fluconazole (Diflucan) in patients with denture stomatitis. The study was perfomed using an open test method, without a comparative drug. Thirty five patients, aged between 28 to 59 years old, were included in the investigation. Candida was cultured from the surface of maxillary dentures and mucosa of all the patients. Patients were treated with oral fluconazole (50mg) given once daily for 14 days and medication of dentures with the same agent. The effect of treatment was evaluated by changes in the clinical erythema, the yeast colonization obtained by mycologic sampling on the palatal mucosa and on the denture base. Results showed that Candida albicans was the most frequent species found in all the patients. We could also demonstrate that the implemented therapy was significantly effective in reducing the candidal populations of the maxillary denture as assessed by scoring of smears and cultures of the palatal denture-bearing mucosa. We concluded that fluconazole is an effective treatment for denture stomatitis, including those patients who persist in wearing dentures continuosly.

Genotipos vacA de Helicobacter pylori en una población venezolana / Helicobacter pylori vac A genotypes in a Venezuelan population

La infección por Helicobacter pylori está  asociada con gastritis, úlcera gastroduodenal, linfoma tipo Maltoma, y es considerado un importante factor de riesgo para el desarrollo de cáncer gástrico. El objetivo de este estudio fue caracterizar los genotipos vacA de cepas de  H. pylori provenientes de biopsias gástricas de una población venezolana. Se evaluaron 128 pacientes con indicación de endoscopia, por enfermedad de las vías digestivas superiores. Se obtuvieron biopsias gástricas de cada uno de ellos para la amplificación de glm y tipificación de las formas alélicas  de vacA, empleando la reacción en cadena de la polimerasa. Los resultados demostraron que 82 (64%) de las muestras fueron positivas para la amplificación de glm. De estos, 51 (62%) de las cepas tenían el genotipo vacA, forma alélica S1 (17 s1a, 29 s1b, 5 s1a+s1b, 10 fueron no tipificables) y 21 (26%) tenían el subtipo S2. El análisis de la región media de vacA reveló que 42 (51%) fueron vac A m1, 26 (32%) m2 y 14 (17%) no tipificaron para m1  o m2. Los resultados de la presente investigación demostraron una alta frecuencia de infección por  H  pylori  genotipo vac A variante alèlica s1/ m1.

Helicobacter pylori infection is associated with gastritis, gastroduodenal ulcer, and Maltoma type lymphoma, and is also considered an important risk factor for the development of gastric cancer. The purpose of this study was to characterize vacA genotypes of H. pylori strains from gastric biopsies from a Venezuelan population. One hundred and  twenty eight patients who required endoscopy due to disease of the upper digestive system were evaluated. A gastric biopsy was taken from each of them for glm amplification and typing of the allelic vacA  forms, using the polymerase chain reaction assay. Results showed that 82 (64%) of the samples were positive for glm amplification. Of these, 51 (62%) of the strains had the vacA genotype, S1 allelic form (17 sla, 29 slb, 5 sla+slb, 10 were not typable) and 21 (26%) had the S2 subtype. The analysis of the vacA mid region revealed that 42 (51%) were vacA m1, 26 (32%) m2, and 14 did not type for m1 or m2. The results of this investigation showed a high frequency of H. pylori vacA genotype infections, s1/m1 allelic variants.

Expresión de moléculas de adhesión y subpoblaciones linfocitarias en lesiones de pacientes con Estomatitis Aftosa Recurrente

Introducción: La Estomatitis Aftosa Recurrente (EAR) es una condición muy común, caracterizada por la recurrencia de úlceras en la mucosa bucal no queratinizada. Actualmente las investigaciones dirigidas a determinar la etiología de la EAR, se enfocan hacia el componente inmunológico, la evidencia más relevante indica que los individuos afectados, presentan alteración de la respuesta inmunológica mediada por células. Adicionalmente, existe atención en destacar la importancia y participación de las moléculas de adhesión en el reclutamiento del infiltrado inflamatorio en esta condición. Objetivo: Determinar la expresión de las moléculas de adhesión ICAM-1, VCAM-1 y las subpoblaciones de linfocitos T CD4+ y CD8+, en lesiones de pacientes con EAR. Material y Método: Se incluyeron 19 pacientes de los cuales 15 presentaban EAR Menor y 4 tenían EAR Mayor. El grupo control incluyó muestras de encía de 10 pacientes sin historia de EAR. A los pacientes les fue realizada una biopsia de la lesión que fue procesada por inmunohistoquímica. Resultados y Conclusión: Se observó un marcado aumento en las subpoblaciones de linfocitos, especialmente de los CD8+, lo que sugiere la implicación de los linfocitos T, y de una respuesta inmune mediada por células activadas, en la patogénesis de esta entidad. La expresión de las moléculas de adhesión de los pacientes con EAR, fue mayor en comparación con el grupo control, siendo estas diferencias estadísticamente significativas. Con respecto a ICAM-1, se observó un fuerte marcaje en el corion-infiltrado seguido del corion-endotelio y epitelio, mientras que en relación a VCAM-1 fue similar en corion-infiltrado y corion-epitelio. Estos resultados implicarían a las moléculas de adhesión jugando un papel importante en las interacciones celulares y en el reclutamiento del infiltrado celular inflamatorio, pudiéndose convertir en blanco para el daño citotóxico y contribuyendo de esta forma, a la ruptura del tejido que ocurre en los sitios de ulceración.

Recurrent aphthous stomatitis (RAS) is the most common oral mucosal disease characterized by aphthous ulcerations in the oral mucosal. Actually the investigations have been directed to determine the etiology of the RAS with special attention to the cellular immune response. Aditionally, is very important to determine the rol of the adhesion molecules in this condition. The purpose of this work was to determine the of the adhesion molecules expression, ICAm-1, VCAM-1, CD4 and CD8 cell count in patients with RAS. We evaluated 19 patients, 15 with RAS minor and 4 with major RAS. The control group included oral health tissue of subjects without RAS. We observed a higher level of CD8 cell count that suggest the implication of T cells and cellular immune response in the pathogenesis of this entity. The adhesion molecules expression was higher than in the control group. These results suggest that the adhesion molecules may play an important role in the cell interactions that we observed in this entity.

Deteccion de Mycobacterium tuberculosis por métodos microbiologicos convencionales y caracteristicas del periodonto de un grupo de pacientes con tuberculosis

Objetivo: establecer la importancia del diagnóstico microbiológico de la Tuberculosis. Metodología: 15 pacientes que asistieron al servicio del Hospital Simón Bolívar, Caracas Venezuela, con cuadro clínico presuntivo de Tuberculosis pulmonar fueron estudiados a través del examen en cavidad bucal, radiografía de tórax, Baciloscopía (BK) y método de Petroff, tomando en consideración los signos y síntomas de cada uno de ellos. Resultados: 11 de las 15 muestras de esputo fueron BK (+) y 4 BK (-). Mycobacterium tuberculosis se aisló en 7 de las muestras incluyendo los pacientes que presentaron BK (-). Desde el punto de vista radiográfico, 10 pacientes presentaron lesión bilateral, 4 lesión unilateral y 1 fibrotorax. El examen en cavidad bucal permitió observar enfermedad periodontal de leve a moderada en 14 de los pacientes, 8 de ellos de manera localizada y 6 de manera generalizada. La detección de M. tuberculosis a través del BK sigue siendo un método de importancia para establecer en conjunto con los antecedentes, epidemiología, clínica, radiografía, serología e histopatología del paciente el diagnóstico definitivo de la tuberculosis, sin embargo, la técnica de PCR puede complementar dicho estudio.

Mycobacterium tuberculosis, aethiological agent of tuberculosis is a curved, aerobic, non sporulated and non motile rod that is a resistant aganaist dissecation effects. 15 patients were selected in this study who assited to Neumonology service located at Simón Bolívar Hospital, Caracas, Venezuela. These patients who had presuntive lung tuberculosis were studied considering their sings and simthomps. Oral cavities of each patient were studied considering their sings and sinthomps. Oral cavities of each patient were examinated and thorax Rx, Bacilloscopy (BK) and Petroff method were taked from each one in order to detect this rod. Our findings related that 11 out of 15 sputum samples were BK (+) and the other 4 were BK (-). M. tuberculosis was isolated in 7 samples, including BK (-) patients. Radiografically, 10 patients presented bilateral lesion, 4 presented unilateral lesion and 1 fibrothorax. Examination of oral cavities showed that 8 patients presented localizated periodontal disease and 6 presented generalizated periodontal disease. Detection of M. tuberculosis using BK continue to be an important method for identification of this bacteria, considering also epidemiological, clinical, radiographical, serological and histopathological studies in order to establish definitive diagnosis of TBC. Aditionally, Polimerase Chain Reaction (PCR) is an excellent method used to complement identification studies of this microorganism.

Periodontal conditions and distribution of Prevotella intermedia, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans in HIV-infected patients undergoing anti-retroviral therapy and in an HIV-seronegative group of the Venezuelan population.

The aim of this study was to determine the periodontal conditions and the distribution of Prevotella intermedia, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans in a group of HIV-infected patients undergoing anti-retroviral therapy (HAART) and in an HIV-seronegative group. The study sample comprised thirty-two (32) HIV positive patients distributed in two groups (11 HIV+ without HAART and 21 HIV+ with HAART) and 16 HIV seronegative patients. Plaque index, gingival index, pocket depth, and clinical attachment level were evaluated at six sites per tooth in all teeth. Subgingival plaque samples were collected from one tooth per quadrant with pocket depth > 4 mm and attachment level > 5 mm. and then analyzed by PCR. The mean value of PI, GI, and CAL of the HIV-infected patients undergoing or not HAART- and the control group were similar the PD was higher in the control group. LGE was observed only in the HIV-infected group and NUP in the HIV+ without HAART therapy. The control group and the total HIV-infected patients showed similar CPG and CPL values. P. intermedia was the most frequently recovered microorganism in all the groups evaluated. The second pathogen with higher prevalence was A. actinomycetemcomitans, P. gingivalis was observed only in one (5%) HIV+ patient under HAART and in three patients (19%) in the control group. The periodontal indexes was not related with the CD4+ count and viral load. Changes observed in the periodontal tissues of patients infected with HIV are similar to those observed in HIV negative subjects.

La placa dental como reservorio de Helicobacter pylori / Dental plaques as Helicobacter pylori reservoirs

Resumen La placa dental ha sido propuesta como un reservorio para Helicobacter pylori, sugiriéndose que el microambiente oral pueda ser un nicho permanente para la bacteria. El objetivo de este estudio fue determinar el papel que juega la placa dental como reservorio de esta especie, evaluando la presencia de ADN de H. pylori en la placa dental y biopsias gástricas, mediante la reacción en cadena de la polimerasa (PCR por sus siglas en inglés). Fueron evaluados 71 pacientes sintomáticos gastrointestinales referidos para examen endoscópico, provenientes del Servicio de Gastroenterología del Hospital Universitario de Caracas, Venezuela y 40 sujetos asintomáticos. Las muestras de placa dental fueron analizadas por PCR, basada en la secuencia del gen ure c. Se tomaron biopsias gástricas para análisis histopatológico y PCR. H. pylori. fue detectado en las biopsias gástricas de 48(68%) de los 71 pacientes, todos con gastritis crónica y en placa dental de 13(18%) de los 71 sujetos. En 8 (17%) de estos 48 pacientes, H. pylori fué también detectado en placa dental. De estos pacientes 4 presentaban adicionalmente displasia y 4 metaplasia. Tres pacientes del grupo control fueron positivos por PCR para H. pylori en placa dental. La placa dental puede ser un reservorio para la bacteria, y su presencia podría representar un factor de riesgo para la reinfección gastrointestinal, posterior al tratamiento de erradicación de la bacteria.

Abstract Dental plaques have been proposed as Helicobacter pylori reservoirs, suggesting that the oral microenvironment can be a permanent niche for the bacterium. The objective of this study was to determine the role played by the dental plaque as reservoir for this species, evaluating the presence of H. pylori DNA in dental plaques and gastric biopsies through the polymerase chain reaction (PCR). The evaluation included seventy-one gastro-intestinal symptomatic patients referred for endoscopic examination by the Gastroenterology Service of the Caracas University Hospital, as well as 40 asymptomatic subjects. The dental plaque samples were analyzed by PCR based on the ure c gene sequence. Gastric biopsies were taken for histopathological analysis and PCR. H. pylori was detected in 48 (68%) gastric biopsies from the 71 patients, all with chronic gastritis, and in the dental plaque of 13 (18%) of the same 71 patients. In 8 (17%) of these 48 patients H. pylori was also detected in dental plaque. Four of these patients also presented displasia and metaplasia. Three patients from the control group were PCR positive for H. pylori in dental plaque. Dental plaques can be a reservoir for these bacteria and their presence could represent a risk factor for gastrointestinal re-infection after treatment for eradicating this bacterium.

Prevalence of oral lesions in HIV patients related to CD4 cell count and viral load in a Venezuelan population.

AIM: To determine the prevalence of oral lesions in a HIV+ group of patients, related to CD4 cell count and viral load in a Venezuelan population. MATERIALS AND METHODS: In the present study, we evaluated 75 HIV+ adult patients, attended at the Center of Infectious Diseases, at the Faculty of Dentistry, Central University of Venezuela. Each patient was clinically examined for detection of oral mucosal lesions. In addition, CD4 cell count was determined by flow cytometry, as well as viral load by RT-PCR (Amplicor HIV-RNA, TM test 1.5, Roche). RESULTS: 85% (64/75) of HIV/AIDS patients showed associated HIV lesions. Oral Candidiasis constituted the most common lesion representing a 61% (39/64), followed by Oral Hairy Leukoplakia 53% (34/64); Oral Leukoplakia 34% (22/64), Melanic Hyperpigmentation 38% (18/64); Papilloma 13 (6/64), Lineal Gingival Erythema 8% (5/64); Aphtous Recurrent Stomatitis 5% (4/64) and Kaposi's Sarcoma 5% (3/64). Only one case of the following lesions were represented by Non Hodgkin Lymphoma, Multifocal Epithelial Hyperplasia, Recurrent Herpes, Histoplasmosis and Molluscum Contagiosum. The patients with a viral load of 30.000 copies/mm3 exhibited oral lesions related with HIV, independent of CD4 cell count, although patients with CD4+ levels of 200 cel/mm3 were more susceptible to develop these lesions. CONCLUSIONS: The most common oral lesion was Oral Candidiasis followed by Oral Hairy Leukoplakia, Oral Leukoplakia and Melanic Hyperpigmentation. A high viral load was strongly associated to the oral lesions occurrence independently of CD4+ cell count.

Prevalencia de lesiones bucales en pacientes VIH +, relación con contaje de células CD4+ y carga viral en una población Venezolana / Prevalence of oral lesions in HIV patients related to CD4 cell count and viral load in a Venezuelan population

Objetivo: Determinar la prevalencia de las lesiones bucales en un grupo de pacientes VIH+, su relación con el contaje de linfocitos CD4 y la carga viral, en una muestra de la población venezolanaMateriales y métodos: En el presente estudio se evaluaron 75 pacientes adultos VIH+, provenientes del Centro para la Atención de Pacientes con Enfermedades Infectocontagiosas de la Facultad de Odontología de la Universidad Central de Venezuela. Cada uno de ellos fue examinado clínicamente para la detección de las lesiones presentes en la mucosa bucal. Igualmente se les determinó el contaje de células CD4+ mediante citometría de flujo, así como la carga viral mediante la técnica TR-RCP (Transcriptasa Reversa-Reacción en Cadena de la Polimerasa) (Amplicor HIV-1-RNA, TM test 1.5, Roche).Resultados: 85% (64/75) de los pacientes VIH/SIDA presentaron lesiones asociadas a la infección por VIH. Entre ellas, la Candidiasis constituyó la forma más frecuente representando un 61% (39/64), seguida por la Leucoplasia Vellosa con 53% (34/64), Leucoplasia Bucal 34% (22/64), Hiperpigmentación Melánica 38% (18/64). Papiloma 13% (6/64), Eritema Gingival Lineal 8% (5/64), Estomatitis Aftosa Recidivante 5% (4/64), Sarcoma de Kaposi 5% (3/64). Se presentó un solo caso (2%) de cada una de las lesiones que se enumeran a continuación: Linfoma No Hodgkin, Hiperplasia Epitelial Multifocal, Herpes Labial, Histoplasmosis y Molusco Contagioso. Los pacientes con una carga viral de 30.000 copias/mm3, presentaron lesiones bucales relacionadas con VIH independientemente del contaje de células CD4+, aunque los pacientes con niveles de CD4+ menores a 200 cel//mm3, fueron mas susceptibles a desarrollar estas lesiones.Conclusiones: La lesión oral mas frecuente fue la Candidiasis bucal, seguida por la Leucoplasia vellosa, Leucoplasia bucal, e Hiperpigmentación melánica. Una alta carga viral estuvo fuertemente asociada a la presencia de lesiones, independientemente del contaje de células CD4+

Aim: To determine the prevalence of oral lesions in a HIV + group of patients, related to CD4 cell count and viral load in a Venezuelan population.Materials and methods : In the present study, we evaluated 75 HIV+adult patients, attended at the Center of Infectious Diseases,at the Faculty of Dentistry, Central University of Venezuela. Each patient was clinically examined for detection of oral mucosal lesions. In addition, CD4 cell count was determined by flow cytometry, as well as viral load by RT-PCR (Amplicor HIV-RNA, TM test 1.5, Roche)Results: 85% (64/75) of HIV/AIDS patients showed associated HIV lesions. Oral Candidiasis constituted the most common lesion representing a 61% (39/64), followed by Oral Hairy Leukoplakia 53% (34/64); Oral Leukoplakia 34% (22/64), Melanic Hiperpigmentation 38% (18/64); Papilloma 13 (6/64), Lineal Gingival Erithema 8% (5/64); Aphtous Recurrent Stomatitis 5% (4/64) and Kaposi’s Sarcoma 5% (3/64). Only one case of the following lesions were represented by Non Hodgkin Lymphoma, Multifocal Epithelial Hyperplasia, Recurrent Herpes, Histoplasmosis and Molluscum Contagiosum. The patients with a viral load of 30.000 copies/mm3 exhibited oral lesions related with HIV, independent of CD4 cell count, although patients with CD4+ levels of 200 cel/mm3 were more susceptible to develop these lesions.Conclusions: The most common oral lesion was Oral Candidiasis followed by Oral Hairy Leukoplakia, Oral Leukoplakia and Melanic Hyperpigmentation. A high viral load was strongly associated to the oral lesions occurrence independently of CD4+ cell count

[Evaluation of K-ras 12 oncogene mutations in Venezuelan patients with Helicobacter pylori infection]. / Evaluación de las mutaciones del oncogen K-ras 12 en pacientes venezolanos infectados con Helicobacter pylori.

This work evaluated the infection of H. pylori in the different gastric pathologies and its association with the oncogen K-ras 12. Endoscopy was performed in 62 patients and 3 biopsies from the antral region were taked and used for the histological diagnostic, PCR, and point mutations determination. The results showed a high incidence of H. pylori infection in patients with active chronic gastritis (AcCG) 90%, chronic atrophic gastritis (AtCG) 70%, intestinal metaplasia (IM) 67%, dysplasia (D) 83%, and decrease in in gastric cancer (GC) 33%. Evaluation of the oncogen K-ras 12 showed that 68% of the patients presented mutations in the different analyzed amino acids. In the 12 codon of the K-ras gene, we observed simple point mutations and combination in the same sample in different gastric pathologies. In AcCG samples were detected the greater number of mutations. A decrease of the point mutations were observed in the progression stages to gastric cancer. The presence of these specific mutations would be tumor markers and it determine the possible development of gastric tumors.