Cytokine and metalloproteinases in gingival fluid from patients with chronic periodontitis.

The purpose of the present research was to determine the levels of IL- 1a, IL- 1 P, TNF-a, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and EMP-8 in gingival crevicular fluid (GCF) of subjects with chronic periodontitis. Clinical measurements were carried out in 20 patients with chronic periodontitis and 11 periodontally healthy controls. The clinical indexes evalua- ted were: gingival index (GI), plaque index (PI), bleeding on probing (BOP), probing depth (PD) and clinical attachment loss (CAL); the measurements were taken at six sites per tooth in all teeth in each subject. GCF samples were taken from one tooth per quadrant, and the levels of mediators were measured using an ELISA test. Statistically significant differences were observed between patients and control group in relation to all clinical parameters evaluated (p<0.05). The gingival concentrations, in pg/mL, of IL-la (patients: 239.06 ± 65.5 vs control: 97.79 ± 15.81), IL-10 (patients: 157.19 ± 36.4 vs control: 63.44 ± 19.04), TNF-a (patients: 10.87 ± 1.7 vs control: 1.15 ± 0.84), IL-6 (patients: 3.77 ± 1.7 vs control: 0.43 ± 0.22), IL-6Sr (patients: 655.59 ± 185.8 vs control: 73.59 ± 23.18), IL-8 (patients: 496.3 ± 155.3 vs control: 206.13 ± 46.63), IL-10 (patients: 10.75 ± 3.6 vs control: 2.41 ± 0.57), MMP-3 (patients: 3531 ± 1558.2 vs control: 724.84 ± 289.51) and MMiP-8 (patients: 8231.70 ± 1279.2 vs control: 1534.67± 814.90) were significantly greater in patients with periodontal disease than in the control group (p<0.00 1). The higher levels of the cytokines and metalloproteinases obtained in this study were significantly associated with the severity of the periodontal disease.

[MMP-3 and MMP-8 levels in patients with chronic periodontitis before and after nonsurgical periodontal therapy]. / Niveles de MMP-3 y MMP-8 en pacientes con periodontitis crónica antes y después del tratamiento periodontal no quirúrgico.

Immune-inflammatory processes are trigged in chronic periodontitis (CP), where matrix metalloproteinases (MMPs) are released and involved in the degradation of the extracellular matrix components that can be detected in the gingival crevicular fluid (GCF). The purpose of the study was to determine the levels of MMP-3 and MMP-8 in GCF, before and after nonsurgical periodontal treatment (NSPT), to evaluate disease activity and therapy response. Eleven patients with PC and eleven healthy controls were selected. Clinical measurements to evaluate gingival index (GI), plaque index (PI), probing depth (PD) and clinical attachment loss (CAL) were made in all the teeth of each individual and in six sites per tooth. GCF samples were taken from one tooth per quadrant, with a pocket depth > or =4 mm and a clinical attachment loss > or =5 mm, and the levels of MMP-3 and MMP-8 measured using an ELISA test. Statistically significant differences in clinical parameters were observed (p < 0.05) between patients with CP and control groups before the periodontal treatment, with significant decrease in all indexes after the NSPT. The initial concentrations of MMP-3 and MMP-8 were significantly higher than those obtained after the NSPT and in the control group, without observing a correlation between the clinical parameters and the levels of MMPs. Increased levels of MMP-3 and MMP-8 in the GCF of patients with PC declined significantly after NSPT, and the difference between the levels in healthy individuals and patients, suggests the important participation of these MMPs in tissue destruction in PC disease..

Human papillomavirus (HPV) type distribution in cervical carcinoma, low-grade, and high-grade squamous intraepithelial lesions in Venezuelan women.

OBJECTIVE: Cervical cancer is an important cause of mortality among women in developing countries, especially in the Latin America and Caribbean (LAC) region. Infection with high-risk (HR) human papillomavirus (HPV) has been identified as the primary cause of cervical cancer. The aim of this study was to determine the frequency of HR-HPV genotypes in low-grade and high-grade squamous intraepithelial lesions (LSIL, HSIL) and cervical carcinoma (CC) among Venezuelan women. MATERIALS AND METHODS: Subjects with histopathological diagnosis of LSIL, HSIL, and CC (LSIL=200; HSIL=100; CC=150) were enrolled in the study after obtaining informed consent. Biopsy samples of these subjects were analyzed to determine the lesion type. HPV detection and typing was done using polymerase chain reaction (PCR) and reverse hybridization. HPV type specific prevalence was determined in subjects with single and multiple infections. RESULTS: HPV DNA was detected in 68%, 95%, and 98.7% of LSIL, HSIL, and CC cases, respectively. HR-HPV and low-risk oncogenic HPV (LR-HPV) was observed in 66.9%/11.8% of LSIL cases, 87.3%/3.2% of HSIL cases, and 91.2%/0.7% of CC cases. HPV types -16/-18 (65%) were the most common high-risk HPV types observed, followed by types -52, -33, -45, and -31. CONCLUSION: Cervical cancer burden in Venezuelan women is substantial. HPV types -16/-18 were the most common types prevalent among Venezuelan women followed by types -52, -33, -45, and -31 (prevalence, ~90.1%). The results of this study provide baseline information on the HPV type distribution, which may facilitate the development of a cervical cancer prevention and control program in Venezuela.

[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.

Comparative analysis of three methods for HPV DNA detection in cervical samples.

High risk HPV infection is considered to play a central role in cervical carcinogenesis. HPV DNA testing has shown to be a very useful tool for screening and following cervical infections. The aim of this study was to compare three methods for HPV DNA detection, along with cytology and colposcopy analysis. Cervical samples were collected from 100 sexually active women in Mérida, western Venezuela. HPV infection was screened using Hybrid-Capture 2 (HC2), L1-Nested-PCR and E6/E7-PCR assays. 40% of the samples (40/100) were HPV positive by at least one of the DNA detection methods. HC2 detected HPV in 12% specimens. L1- and E6/E7-PCRs showed 50% sensitivity and 77% specificity.The agreement rate between HC2 and both PCR assays was 65%. Kappa value showed moderate concordance between HC2 and both PCR methods (kappa=0.55; CI 95%). Also moderate concordance was seen when L1- and E6/E7-PCRs were compared (kappa=0.48; CI 95%). There was a significant association between the Schiller test and E6/E7-PCR (p=0.006) for HPV infection. An acceptable agreement between all three assays for HPV detection was observed. Nevertheless, different PCR formats need to be further analyzed in order to make the right choice of method for HPV testing.

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.

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.

Presence of human papillomavirus infection determined by hybrid capture assay in cervical lesions in a Venezuelan population.

The aim of this study was to establish the presence of HPV infection in cervical lesions in a Venezuelan population, by Hybrid Capture Assay II (HCA II), and its association with cytological diagnosis. The study included 1483 cervical samples analysed at Laboratorios CITOMED, Caracas, Venezuela, from 2005 to 2007. The woman age range was between 20 and 58 years, and the mean age was 28.8. HPV infection was determined using HCA II. The cytological diagnosis of the smears showed LSIL in 1120/1483 samples (75.5%), HSIL in 354/1483 (23.9%) and ASC-US, in 9/1483 (0.6%). The positivity of HPV DNA detected by HCA II was 54.6% (811/1483). Of the positives cases, 138/811 (17%) presented HPV DNA of low oncogenic risk and 673/811 (82.9%) had high-risk HPV. There were significant differences in the low and high oncogenic HPV type frequencies of the evaluated samples (p > 0.0001). Low risk HPV types were detected in 127 cases of LSIL, 9 of HSIL and 2 of ASC-US. High-risk HPV was detected in most of the cases: 361 LSIL, 308 HSIL and 4 ASC-US. Our study showed a high presence of cervical infection by human papillomavirus of a high risk genotype. Our results contribute to the epidemiological data that report diversity in the prevalence rates in different countries.

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.

Association of viral load and physical status of HPV-16 with survival of patients with head and neck cancer.

INTRODUCTION: Head and neck cancers (NHCs) are of multifaceted origins, and tobacco and alcohol are the primary risk factors. Currently, other factors associated with the genesis of these tumours are being considered, among these viral infections, especially human papillomavirus (HPV) infection. OBJECTIVE: The objective was to evaluate HPV infection, HPV-16 E6 load and its physical status in patients with squamous cell carcinoma in the head and neck and evaluate its effects in the survival of these patients. METHODOLOGY: A total of 80 fresh biopsies of HNC were evaluated. The genetic material was extracted using the commercial kit QIAGEN. The detection and classification of HPV were carried out using INNO-LiPA, whereas the quantification and analysis of integration of the viral genome into the host cell were carried out using real-time PCR. RESULTS: The average age of the patients included was 60.34 ± 14.48 years, with a predominance of the male gender. The most frequent HPV infection was genotype 16 (52.8%), with an average of 10 copies of the HPV-16 E6/ß-globin gene. Furthermore, an integration of the viral genome in the host cell was observed in 86% of cases with a statistically significant relationship between the location of the tumour and the viral load (p < 0.05). CONCLUSIONS: HPV-16 is the most common infection, and its physical status in the host cell is the determining factor in establishing response to treatment. However, more studies are needed to demonstrate the role of HPV infection in carcinogenesis.

Oral and oropharyngeal cancer in a Venezuelan population.

The aim of this work was to analyze diagnosed cases of Oral Cancer (OC) and Oropharyngeal Cancer (OPC) in a Venezuelan population. We clinically evaluated 130 patients with OC and OPC and a histopathologic diagnosis of squamous cell carcinoma. The patients were analyzed according to gender age, and use of alcohol and tobacco and the tumors were classIfied based on anatomic location, staging parameters, and degree of difFerentiation. Ninety one patients (70%) were male and 39 (30%) were female. Patients' age ranged from 26 to 86 years old. Use of smoking tobacco, alcohol or both was reported by 84.3%, 49.1% and 45.4% of patients, respectively, and was more frequent in males. The most common oropharyngeal anatomic location was the base of the tongue (22.3%), followed by the tonsils (13.9%), while the most frequently affected oral location was the oral tongue (19.2%) followed by the gingiva and alveolar mucosa (10.8%), and the floor of mouth (7.7%). The majority of tumors (77.7%) were diagnosed at an advanced stage (Stage III or IV); metastasis to the regional lymph nodes occurred in 53.1% of cases. According to degree of diferentiation, well, moderately and poorly difFerentiated tumors accounted for 45.4%, 46.1% and 8.5% of cases, respectively. Well differentiated tumors accounted for 56.7% of OC cases, while the majority of OPC cases were classified as moderately or poorly differentiated (72.3%) (p < or = 0.002). Also, non-metastatic cases (NO) showed a predominance of well-diferentiated tumors (61.2%), while metastatic tumors (N+) were classified as moderately or poorly differentiated in 89.8% of cases (p < or = 0.0001). Our study population was characterized by a predominance of smokers and/or drinkers and a predilection for male patients. Most tumors were diagnosed at an advanced stage with a high incidence of metastatic spread to the regional lymph nodes, indicating possible delays in diagnosis. Less differentiated tumors were more frequently encountered among OPC cases and accounted for the vast majority of metastatic cases, supporting the prognostic value of assessing the degree of difFerentiation.

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.

Lymphocyte subsets predictive value and possible involvement of human papilloma virus infection on breast cancer molecular subtypes.

AIM: To detect human papilloma virus (HPV) presence and to characterize cellular immune response in breast cancer patients. METHODS: A total of 74 women were included, of which 48 samples were from patients diagnosed with breast cancer and 26 patients with benign pathology of the breast. Molecular subtype classification was performed based on the immunohistochemical reports of the tumor piece. HPV genome detection and genotyping from fresh breast biopsies was performed using the INNO-LIPA HPV Genotyping Extra test (Innogenetics, Ghent, Belgium). CD3+, CD4+, CD8+ and natural killer (NK)+ cells levels from peripheral blood samples from patients with breast cancer and benign pathology were measured by flow cytometry. RESULTS: Luminal A was the most frequent breast cancer molecular subtype (33.33%). HPV was detected in 25% of the breast cancer patients, and genotype 18 was the most frequent in the studied population. The mean of CD3+, CD4+ and CD8+ subpopulations were decreased in patients with breast cancer, in relation to those with benign pathology, with a statistically significant difference in CD8+ values (P = 0.048). The mean of NK+ cells was increased in the benign pathology group. The average level of CD3+, CD4+, CD8+ and NK+ cells decreased as the disease progressed. HER2+ and Luminal B HER2+ tumors had the lowest counts of cell subsets. HPV breast cancer patients had elevated counts of cellular subsets. CONCLUSION: Determining level changes in cellular subsets in breast cancer patients is a useful tool to evaluate treatment response.

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.

[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.

[Importance of IgG anti-CagA antibodies of Helicobacter pylori in Venezuelan patients with gastric diseases]. / Importancia de la respuesta humoral de IgG anti-CagA de Helicobacter pylori en pacientes venezolanos con enfermedades de las vías digestivas superiores.

Helicobacter pylori is one of the most common bacterial infections worldwide. It is associated with chronic gastritis, peptic ulcer disease and constitutes a major risk factor for gastric adenocarcinoma and lymphoma. The aim of this study was to evaluate the specific serologic immunoglobulin G (IgG) response to whole cells proteins, CagA and urease antigens of Helicobacter pylori in a Venezuelan population. We evaluated 66 patients from the Hospital Universitario de Caracas, attending in the gastroscopy service. H. pylori infection was detected by culture and rapid urease test. IgG antibodies against, CagA and ureases were tested by enzyme-linked immunosorbent assay method using highly purified recombinant antigens. We demonstrated the presence of H. pylori in 48/66 (72.7%), by culture and rapid urease test. We found a seroprevalence of 45 (68%) to whole cells, 34/66 (51%) to CagA and 18/66 (27%) to urease. The positive rates of CagA antibodies in patients with gastric ulcer, gastric cancer and chronic gastritis were 87.8%, 77.7% y 40.8% respectively. The serum antibodies anti-CagA were similar between peptic ulcer disease and gastric cancer patients.

Presence of human papillomavirus in breast cancer and its association with prognostic factors.

Breast cancer accounts for 16% of all female cancers worldwide, and in Venezuela, it is the leading cause of death among women. Recently, the presence of high-risk genotypes of human papillomavirus (HPV) has been demonstrated in breast cancer and has been associated with histopathological features of the tumours. In Venezuela, there is no study which determines the association between the presence of HPV in breast cancer and the histopathological features. The aim of this investigation is to evaluate the presence of HPV in the different types of breast cancer, according to their molecular classification, based on the expression of ER, PR, HER2 and Ki67. With this purpose in mind, we assessed the presence of the HPV genome in 24 breast cancer samples diagnosed with infiltrating ductal carcinoma, ductal carcinoma in situ (DCIS) and lobular carcinoma, by the INNO-LIPA genotyping extra kit and the evaluation of the markers ER, PR, HER2, and Ki67 by immunohistochemistry. The viral genome was found in 41.67% of the total number of samples, 51 being the most frequent genotype with 30.77%, followed by types 18 and 33, with 23.08%, respectively. Most tumours were found in the group of luminal A, with a low range of Ki67 expression. The presence of HPV in breast tumours could affect their growth pattern and metastatic power.

Evaluación de virus papiloma humano de alto y bajo riesgo oncogénico en la cavidad bucal de pacientes VIH positivos / Evaluation of human papillomavirus of high and low oncogenic risk in the oral cavity of HIV positive patients

Objetivo. Realizar la detección y tipificación de virus papiloma humano (VPH) en la cavidad bucal de un grupo de pacientes VIH positivos, atendidos en el Centro de Atención de Pacientes con Enfermedades Infectocontagiosas (CAPEI) de la Facultad de Odontología-Universidad Central de Venezuela. Métodos. Se evaluaron 31 hisopados bucales de pacientes VIH positivos para la infección por VPH, mediante el Sistema de Hibridación Reversa de INNO-LiPA que detecta 28 genotipos de VPH de alto y bajo riesgo oncogénico. Resultados. El 61,0% de las muestras evaluadas presentó infección por VPH. El genotipo 6 fue el más frecuente (73,68%), seguido de los genotipos 18, 11 y 16 con 63,16%, 37,0% y 32,0% respectivamente. El 74,0% de las muestras positivas para VPH presentaron infecciones múltiples, siendo más frecuente la coinfección mixta (35,70%) con los genotipos de VPH-6/18 de bajo y alto riesgo oncogénico, 21,40% con los genotipos 6/11/18 y 6/11, cada una. Seguido de 14,30% de las muestras que presentaron infección con VPH-6/11/16/18 y 7,10% con los genotipos 11/16 de bajo y alto riesgo oncogénico. Conclusiones. La alta frecuencia de infección con VPH de alto riesgo oncogénico y la presencia de múltiples genotipos observada en la cavidad bucal de individuos VIH positivos, que no presentaron lesiones compatibles con esta infección en el examen extra e intrabucal, indica que los métodos moleculares de diagnóstico son importantes en la detección de infecciones subclínicas y latentes, lo que puede permitir un mejor seguimiento y manejo más oportuno de estos pacientes con mayor riesgo de desarrollar neoplasias malignas en la cavidad bucal.

Objective. Perform human papilomavirus (HPV) detection and typing in the oral cavity in a group of HIV positive patients, treated at the Center for Care of Patients with Infectious Diseases (CAPEI) of the Faculty of Dentistry-Central University of Venezuela. Methods. Thirty-one oral swabs from HIV-positive patients were evaluated to detect HPV infection using the INNO-LiPA Reverse Hybridization System that detects 28 HPV genotypes of high and low oncogenic risk. Results. The 61.0% of the evaluated samples had an HPV infection. The low risk genotype 6 was the most frequent (73.68%), followed by genotypes 18, 11 and 16 with 63.16%, 37.0%, and 32.0%, respectively. Seventy-four percent of HPV positive samples had multiple infections, being the more frequent the mixed coinfection (35.70%), with HPV genotypes 6/18 of low and high oncogenic risk, 21.40% with genotypes 11/6/18 and 6/11, each one, followed by 14.30% of the samples presented infection with HPV-6/11/16/18 and 7.10% with the 11/16 genotypes of low and high oncogenic risk. Conclusions. The high frequency of infection with high oncogenic risk HPV types and the presence of multiple genotypes observed in the oral cavity of HIV positive patients, who did not present lesions compatible with this infection in the extra and intraoral examination, indicates that molecular diagnostic methods are important in the detection of subclinical and latent infections, which may allow better follow-up and more timely management of these patients with a higher risk of developing malignant lesions in the oral cavity

Persistent infection with high-risk human papilloma viruses: cohort study, Mérida, Venezuela.

Cervical lesions have been associated with infection by high-risk human papilloma virus (high-risk HPV). In 409 women aged >15 years high-risk HPV lesions were identified. In a cohort of this population persistent infection was compared with cytological, colposcopic, and histological lesions. Cervical scrapes were taken and DNA was isolated. HPV was detected by PCR in the E6/E7 region. Genotyping was performed by PCR nested multiple E6/E7. HPV was detected in a 37.40% (153/409), high-risk HPV in 86% (153/178), HPV18 46.64% (83/178), HPV16 34.28% (61/178). Among these 53.93% (96/178) were multiple infections, and HPV18/16 (30/96) was the most frequent 31.25%. The cytology showed changes in 15% of positive patients. A 49.67% in women positive for HPV infection showed abnormalities in the colposcopic study, a relationship that turned out to be statistically significant ( p < 0.0019 test χ(2)). Among all 85% of the women were younger than 45 years of age. Fifty-seven patients were evaluated 15 months after the base study, with initial prevalence of morbidity 49.12% (28/57) and at the end 10.53% (6/57), showing in 89.29% (25/28) negative for HR-HPV infection, 10.34% (3/28) showed persistence of infection, 17.54% (10/57) presented cytological alterations, with 80% of positivity for HPV, and a regression of 100% (10/10) of the previously identified lesions. With colposcopy, 50% (14/28) presented alterations related to HPV, of these 85.71% (12/14) showed regression of such an alteration. The cumulative incidence for HPV was 10.34% (3/29). The incidence rate was 4.23% (3/71), which is equal to 4.23 new cases of HPV infection per 100 people, per year of follow-up. In conclusion, the present work shows a high frequency of infection by high-risk HPV, with predominance of HPV18 and 16 and in general for multiple infections. Colposcopy was better predictor than the Pap smear for infection. The follow-up study revealed a low percentage of persistent infection, and a high frequency of negativity for viral infection, high regression of cytological and colposcopic lesions, a low cumulative and incidence rate similar to that reported by other Latin American countries and higher than the European countries.

Detection of Helicobacter pylori DNA in the oral cavity and gastroduodenal system of a Venezuelan population.

Dental plaque has been suggested as a reservoir for Helicobacter pylori but the hypothesis that the oral microflora may be a permanent reservoir of H. pylori is still controversial. The aims of this study were to determine the presence of H. pylori DNA in the gastric antrum and dental plaque of a Venezuelan population by PCR and to investigate the relationship between this infection and the oral hygiene index. Thirty-two patients from the Hospital Universitario de Caracas, attending for routine gastroscopy, and 20 asymptomatic subjects (control group) were evaluated. The patients' gingiva and plaque were assessed by the gingival and plaque indices of Sillness and Löe. Supragingival plaque was analysed by a PCR for a specific internal urease gene. Gastric antrum biopsies were taken for histological examination and PCR. H. pylori was detected in antral samples from 24 (75%) of 32 patients, all of whom had chronic gastritis. H. pylori was also detected in dental plaque samples of 12 (37.5%) of the 32 patients. In 7 (58%) of these 12 patients, H. pylori was identified in the gastric biopsy. Seven patients with chronic gastritis carried H. pylori in dental plaque and antral samples. Of these patients, four also had dysplasia and one had metaplasia. Three subjects in the control group were positive by PCR. In the present study there was no correlation between H. pylori infection and dental hygiene, dental caries, periodontal disease or use of dentures. The oral cavity may be a reservoir for H. pylori infection and oral secretions may be an important means of transmission of this micro-organism. H. pylori in dental plaque may represent a risk factor for gastrointestinal re-infection and ulcer relapse after antibiotic therapy.