Paulo Roberto Eleutério de Souza

Immunological's host profile for HPV and Chlamydia trachomatis, a cervical cancer cofactor.

Over 100 different genotypes of human papillomavirus (HPV) have been isolated to date, while Chlamydia trachomatis is the most common bacterial sexually-transmitted pathogen. This review considers evidence that C. trachomatis infection became a cofactor for HPV establishment and the development of cervical intraepithelial neoplasia.

Prevalence of autoimmune thyroid disease and thyroid dysfunction in young Brazilian patients with type 1 diabetes.

Abstract:  Patients with an autoimmune condition are known to be at higher risk of developing other autoimmune disorders. Type 1 diabetes may be associated with additional autoimmune disorders including autoimmune thyroid disease. The aim of this study was to investigate the prevalence of thyroid autoantibodies in a group of children, adolescents, and young adults with type 1 diabetes from northeastern Brazil as well as their significance for the development of thyroid disorders. The study design was cross‐sectional and descriptive, analyzing young people with a previous type 1 diabetes diagnosis. Two hundred and fourteen children and adolescents with prior diagnosis of type 1 diabetes were evaluated. Antibodies to thyroperoxidase (anti‐TPO) were determined in all patients and thyroid‐stimulating hormone (TSH) levels. The anti‐TPO antibody test was positive in 54 out of the 214 patients studied, resulting in an overall prevalence of 25.2%. Among the anti‐TPO‐positive subjects, females were predominant (72%) over males (28%) (p < 0.001). A total of 55.5% patients with positive anti‐TPO antibodies had abnormal TSH levels. Clinically significant hypothyroidism was found in 29.6% and subclinical hypothyroidism in 22.2% of patients with positive anti‐TPO. Hyperthyroidism was present in only 3% of them. Our results demonstrate the high prevalence of autoimmune thyroiditis in patients with type 1 diabetes and the need for these patients of regular screening to make a precocious diagnosis of thyroid dysfunction.

IL-18 gene promoter polymorphism is involved in HIV-1 infection in a Brazilian pediatric population

In our study, we identified a polymorphism (C-607A) in the promoter region of the IL-18 gene that shows different frequencies between human immunodeficiency virus (HIV)-1-infected children and healthy controls in a pediatric Brazilian population. The presence of the −607 C allele correlates to HIV-1 infection and confers an increased risk of infection in subjects carrying the single nucleotide polymorphism.

Principles and applications of polymerase chain reaction in medical diagnostic fields: a review

Recent developments in molecular methods have revolutionized the detection and characterization of microorganisms in a broad range of medical diagnostic fields, including virology, mycology, parasitology, microbiology and dentistry. Among these methods, Polymerase Chain Reaction (PCR) has generated great benefits and allowed scientific advancements. PCR is an excellent technique for the rapid detection of pathogens, including those difficult to culture. Along with conventional PCR techniques, Real-Time PCR has emerged as a technological innovation and is playing an ever-increasing role in clinical diagnostics and research laboratories. Due to its capacity to generate both qualitative and quantitative results, Real-Time PCR is considered a fast and accurate platform. The aim of the present literature review is to explore the clinical usefulness and potential of both conventional PCR and Real-Time PCR assays in diverse medical fields, addressing its main uses and advances.

Agreement between Methods for Diagnosing HPV-Induced Anal Lesions in Women with Cervical Neoplasia

Objective: To evaluate agreement between 3 methods for screening anal intraepithelial lesions: anal cytology, anoscopy and human papillomavirus (HPV) detection by PCR. Study Design: This prospective, cross-sectional study screened 324 women with cervical neoplasia for anal neoplasia. Agreement between methods was calculated using the ĸ coefficient. Results: Of 324 anal cytologies performed, 31.5% (n = 102) were found to be abnormal: low-grade anal lesions were detected in 19.1% (n = 62) of cases, high-grade lesions in 3.1% (n = 10) and atypical squamous cells of undetermined significance in 9.3% (n = 30). With respect to the biopsies, 25.7% (n = 20) were positive, consisting of 7 cases of HPV infection, 5 anal intraepithelial neoplasia (AIN) grade 1, 6 AIN grade 2, and 2 AIN grade 3. Twenty-one samples (6.5%) were inadequate for HPV analysis. Of the 303 adequate samples, 84.2% (n = 255) tested positive for HPV. Agreement between cytology and anoscopy was fair (ĸ = 0.31). Agreement between PCR for HPV and cytology was slight (ĸ = 0.08) and no agreement was found between PCR for HPV and anoscopy (ĸ = 0.00). Conclusion: Agreement between the different methods of diagnosing HPV-induced anal lesions is slight to fair; however, anal cytology permits identification of cases in which lesions are present, allowing them to be referred for anoscopy and biopsy.

Evaluation of real time PCR technique to diagnosis of human T-lymphotropic virus type I (HTLV-I) in patients in the Hematologia da Fundação Hemope Hospital, in Northeastern Brazil

As a high degree of homology exists between the proviral genomes of HTLV-I and HTLV-II, there is significant cross-reactivity. Therefore although detection of HTLV antibodies is characteristic of viral infection, it is not sufficient to confirm the presence of the viral type. Molecular tests used to diagnose the HTLV-I/II viruses are based on investigations of proviral genomic sequences, and allow for an infection to be diagnosed prior to the appearance of any sign or symptom. The HTLV proviral load in infected individuals can be determined using real-time PCR, a faster method with less risk of contamination than simple or nested PCR. We analyzed 63 samples from the Hemope Hospital, of which 33 were from HTLV seropositive individuals and 30 from blood donors, to determine the type of virus and the proviral load. The sensitivity of qualitative PCR in comparison to ELISA was 87.5% (95% IC: 70.1 - 95.9%) and the specificity was 100% (IC 95%: 85.9 - 100.0%). The sensitivity and specificity of real-time PCR in comparison to the serological test (ELISA) were 100% (95% IC: 86.7 - 100.0%) and 96.67% (95% IC: 80.9 - 99.8%) respectively. The proviral load in the seropositive individuals ranged from 13 to 343820 copies/106 PBMC cells. Our study also observed that individuals with TSP/HAM had a higher proviral load than those who showed no symptoms. The use of real time PCR for routine clinical testing of infected individuals will play a significant role in identifying the virus type and determining the proviral load, thereby providing more appropriate treatment.

MAO‐B and COMT Genetic Variations Associated With Levodopa Treatment Response in Patients With Parkinson's Disease

The most commonly used Parkinsons disease (PD) treatment is the replacement of dopamine by its levodopa precursor (l‐dopa). Monoamine oxidase‐B (MAO‐B) and catechol‐o‐methyl transferase (COMT) are enzymes involved in the metabolism and regulation of dopamine availability. In our study we investigated the possible relation among selected single‐nucleotide polymorphisms (SNPs) in the MAO‐B (rs1799836) and COMT (rs4680) genes and the therapeutic response to levodopa (l‐dopa). A total of 162 Brazilian patients from the Pro‐Parkinson service of Clinics Hospital of Pernambuco diagnosed with sporadic PD and treated with levodopa were enrolled. PD patients were stratified into 2 groups according to the daily levodopa dose. MAO‐B and COMT SNP genotyping was conducted by polymerase chain reaction–restriction fragment length polymorphism. After multivariate analysis, we observed a significant difference between PD groups for the following variables: sex (P = .02), longer duration of disease (P = .02), longer levodopa therapy duration (P = .01), younger onset of PD (P = .01), and use of COMT inhibitor (P = .02). We observed that patients carrying MAO‐B (rs1799836) A and AA genotypes and COMT (rs4680) LL genotype suffered more frequently from levodopa‐induced‐dyskinesia. In addition, we found an increased risk of 2.84‐fold for male individuals carrying the MAO‐B G allele to be treated with higher doses of levodopa (P = .04). We concluded that before beginning PD pharmacological treatment, it is important to consider the genetic variants of the MAO‐B and COMT genes and the sex, reinforcing the evidence that sexual dimorphism in the genes related to dopamine metabolism might affect PD treatment.

A Fast Method for DEFB1-44C/G SNP Genotyping in Brazilian Patients with Periodontitis

AIM Defensins are cationic antimicrobial peptides expressed in epithelial cells. Such peptides exhibit antibacterial, antifungal and antiviral properties, and are a component of the innate immune response. It has been suggested that they have a protective role in the oral cavity. This study evaluated the DEFB1 polymorphism in diabetic patients with or without periodontitis in comparison to healthy controls. MATERIAL AND METHODS We used Hairpin-Shaped Primer (HP) assay to study the distribution of the -44 C/G SNP (rs1800972) in 119 human DNAs obtained from diabetic patients and healthy control patients. RESULTS The results indicate that there are no differences in distribution between groups and that in diabetic periodontitis patients the homozygous mutant could be found more frequently. CONCLUSION Further studies are necessary in order to investigate the role of DEFB1 polymorphisms in diabetic periodontitis patients and the influence of the peptide in periodontal pathogens.

The influence of SLC6A3 and DRD2 polymorphisms on levodopa-therapy in patients with sporadic Parkinson's disease

The aim of this study was to evaluate a possible relationship between DRD2/ANKK1 (rs1800497) and SLC6A3/DAT1 (rs28363170) gene polymorphisms with the response to levodopa (L‐DOPA)‐therapy in patients with Parkinsons disease (PD).

Mannose binding lectin genes (MBL2) polymorpshisms and the periodontal disease in diabetic patients

PURPOSE: To assess the association between the polymorphism in exon-1 of the MBL2 gene and the periodontal disease in type 2 diabetic patients. METHODS: The sample comprised of 100 patients, who were submitted to a clinical periodontal examination that evaluated in six sites per tooth the probing depth (PD), bleeding on probing (BOP), clinical attachment loss (CAL), plaque index (PI) and the number of teeth present. Periodontal disease was defined as at least four sites with loss of attachment of >5 mm, with one or more of those sites having a pocket of > 4 mm. The collection of scaling cells from the oral mucosa was carried out and the detection of MBL2 polymorphism was made by real time PCR and melting temperature curve analysis. RESULTS: In a type 2 diabetic population, no significant statistical differences in MBL2 polymorphisms genotype or allele frequencies were observed among subjects with periodontal disease. CONCLUSION: This study indicates that the polymorphisms in exon-1 of the MBL2 gene are not related to periodontal disease in a type 2 diabetic population.