Analysis of genotypic variation in genes associated with virulence in Aggregatibacter actinomycetemcomitans clinical isolates

Background and Objective: Although certain serotypes of Aggregatibacter actinomycetemcomitans are associated more with aggressive periodontitis than are other serotypes, the correlation between distinct lineages and virulence traits in this species is poorly understood. This study aimed to evaluate the polymorphism of genes encoding putative virulence factors of clinical isolates, and to correlate these findings with A. actinomycetemcomitans serotypes, genotypes and periodontal status of the hosts. Material and Methods: Twenty-six clinical isolates from diverse geographic populations with different periodontal conditions were evaluated. Genotyping was performed using pulse-field gel electrophoresis. Polymorphisms in the genes encoding leukotoxin, Aae, ApaH and determinants for serotype-specific O polysaccharide were investigated. Results: The isolates were classified into serotypes a-f, and exhibited three apaH genotypes, five aae alleles and 25 macrorestriction profiles. Two serotype b isolates (7.7%), obtained from Brazilian patients with aggressive periodontitis, were associated with the highly leukotoxic genotype; these isolates showed identical fingerprint patterns and aae and apaH genotypes. Serotype c, obtained from various periodontal conditions, was the most prevalent among Brazilian isolates, and isolates were distributed in two aae alleles, but formed a genetically distinct group based on apaH analysis. Cluster analysis showed a close relationship between fingerprinting genotypes and serotypes/apaH genotypes, but not with aae genotypes. Conclusion: Apart from the deletion in the ltx promoter region, no disease-associated markers were identified. Non-JP2-like strains recovered from individuals with periodontal disease exhibited considerable genetic variation regarding aae/apaH genotypes, serotypes and XhoI DNA fingerprints.

Asthma Polymorphic Loci genotyping in Madeira population: identification of potential genetic markers of disease susceptibility, severity and clinical relevance

Asthma is a complex disease, influenced by both environmental and genetic factors. In this study, the analysis of multiple environmental factos assessed by questionnaire and the genotyping of SNPs IL131c.144 G/A, IL41590 C/T, IL41RP2 253183, ADRB21c.16 A/G, ADAM331V4 C/G, ADAM331S1 c.710 G/A, GSDML1236 C/T and STAT6121 C/T were performed in a sample of Madeiran asthmatic patients and their families, and their association to asthma susceptibility and severity was assessed. Family, environmental, social and individual factos such as the presence of rhinitis in one of the parents,the habitation conditions, the family smoking habits, individual food habits and allergen sensitivity, were found to account for asthma severity. IL41590*T and IL41RP2*183$ alleles as well as the combined genotypes IL41590*CT/IL41590*TT and IL41 RP2*253183/IL41RP2*253183 were associated to both asthma susceptibility and severity.GSDML1236*TT was found associated only to asthma severity.Allele ADAM331 V4*C was significantly overM transmitted to asthmatic offspring being linked with the disease by TDT. These findings suggest that in addition to environmental influences, IL41 590 C/T, IL41RP2 253183, ADAM331V4 C/G and GSDML1236 C/T SNPs may constitute important genetic factos contributing to asthmasusceptibility and/or severity in Madeira population.

Non-invasive strategy in assessing asthma through biofluids metabolomics exploration: exhaled breath and urine potentialities

Asthma is a significant health issue in the pediatric population with a noteworthy growth over the years. The proposed challenge for this PhD thesis was the development of advanced methodologies to establish metabolomic patterns in urine and exhaled breath associated with asthma whose applicability was subsequently exploited to evaluate the disease state, the therapy adhesion and effect and for diagnostic purposes. The volatile composition of exhaled breath was studied combining headspace solid phase microextraction (HS-SPME) with gas chromatography coupled to mass spectrometry or with comprehensive two-dimensional gas chromatography coupled to mass spectrometry with a high resolution time of flight analyzer (GC×GC–ToFMS). These methodologies allowed the identification of several hundred compounds from different chemical families. Multivariate analysis (MVA) led to the conclusion that the metabolomic profile of asthma individuals is characterized by higher levels of compounds associated with lipid peroxidation, possibly linked to oxidative stress and inflammation (alkanes and aldehydes) known to play an important role in asthma. For future applications in clinical settings a set of nine compounds was defined and the clinical applicability was proven in monitoring the disease status and in the evaluation of the effect and / or adherence to therapy. The global volatile metabolome of urine was also explored using an HSSPME/GC×GC–ToFMS method and c.a. 200 compounds were identified. A targeted analysis was performed, with 78 compounds related with lipid peroxidation and consequently to oxidative stress levels and inflammation. The urinary non-volatile metabolomic pattern of asthma was established using proton nuclear magnetic resonance (1H NMR). This analysis allowed identifying central metabolic pathways such as oxidative stress, amino acid and lipid metabolism, gut microflora alterations, alterations in the tricarboxylic acid (TCA) cycle, histidine metabolism, lactic acidosis, and modification of free tyrosine residues after eosinophil stimulation. The obtained results allowed exploring and demonstrating the potential of analyzing the metabolomic profile of exhaled air and urine in asthma. Besides the successful development of analysis methodologies, it was possible to explore through exhaled air and urine biochemical pathways affected by asthma, observing complementarity between matrices, as well as, verify the clinical applicability.

Relative abundance and population biology of the non-indigenous crab Charybdis hellerii (Crustacea: Brachyura: Portunidae) in a southwestern Atlantic estuary-bay complex

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Smoking and Periodontal Disease: Clinical Evidence for an Association

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Longitudinal clinical and radiographic evaluation of severely intruded permanent incisors in a pediatric population

Intrusion is defined as the axial dislodgment of the tooth into its socket and is considered one of the most severe types of dental trauma. This longitudinal outcome study was undertaken to evaluate clinically and radiographically severely intruded permanent incisors in a population of children and adolescents. All cases were treated between September 2003 and February 2008 in a dental trauma service. Clinical and radiographic data were collected from 12 patients (eight males and four females) that represented 15 permanent maxillary incisors. Mean age at the time of injury was 8 years and 9 months (range 7-14 years and 8 months). Mean time elapsed to follow-up was 26.6 months (range 10-51 months). The analysis of data showed that tooth intrusion was twice as frequent in males. The maxillary central incisors were the most commonly intruded teeth (93.3%), and falling at home was the main etiologic factor (60%). More than half of the cases (53.3%) were multiple intrusions, 73.3% of the intruded teeth had incomplete root formation and 66.6% of the teeth suffered other injuries concomitant to intrusion. Immediate surgical repositioning was the treatment of choice in 66.7% of the cases, while watchful waiting for the tooth to return to its pre-injury position was adopted in 33.3% of the cases. The teeth that suffered additional injuries to the intrusive luxation presented a fivefold increased relative risk of developing pulp necrosis. The immature teeth had six times more chances of presenting pulp canal obliteration that the mature teeth and a lower risk of developing root resorption. The most frequent post-injury complications were pulp necrosis (73.3%), marginal bone loss (60%), inflammatory root resorption (40%), pulp canal obliteration (26.7%) and replacement root resorption (20%). From the results of this study, it was not possible to determine whether the type immediate treatment had any influence on the appearance of sequelae like pulp necrosis and root resorption after intrusive luxation, but the existence of additional injuries and the stage of root development influenced the clinical case outcome in a negative and positive manner, respectively.

Clinical and Microbiologic Follow-Up Evaluations After Non-Surgical Periodontal Treatment With Erbium:YAG Laser and Scaling and Root Planing

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Distribution and hydrolytic enzyme characteristics of Candida albicans strains isolated from diabetic patients and their non-diabetic consorts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Clinical applications of non-culture based methods for the diagnosis and management of opportunistic and endemic mycoses

Nonculture based methods for the detection of infections caused by fungal pathogens are becoming more important tools in the management of infected patients. Detection of fungal antigens and DNA appear to be the most promising in this respect for both opportunistic and endemic mycoses. In this article we present an overview of the most recent developments in nonculture based methods and examine their value in clinical practice.

A clinical follow-up study of the periodontal conditions of RPD abutment and non-abutment teeth

The purpose of this study was to evaluate the periodontal conditions of removable partial denture (RPD) wearers, comparing direct and indirect abutment teeth, and the teeth not involved in the denture design before denture placement and 1 year later. Fifty patients (32 women and 18 men), average age 45, were assessed by the same examiner at the moment of denture insertion and 3, 6, 9 and 12 months later. The following items were verified in each assessment: probing depth (PD), plaque index (PI) and gingival index (GI). PD and PI data were evaluated by ANOVA test for linear trend followed by Tukey-Kramer post-test, while GI data were analysed by Friedman's test. Results showed that the teeth not involved in the denture design were the least affected for all variables studied. It was also verified that PD and GI mean values increased from the initial assessment to 1 year of RPD wearing in every group, but that only PI showed a significant increase. This study indicated that direct and indirect retainer elements tend to undergo more damaging periodontal effects associated with RPD wearing when compared with non-abutment elements. Plaque index values were significantly higher after 1 year of denture use.

Results of prostate cancer screening in non-symptomatic men

Objectives: To verify prostate cancer prevalence in non-symptomatic men between 50 and 70 years old as well as cancer characteristics. Material and Methods: 2815 non-symptomatic men had total PSA and digital rectal examination performed between March 1998 and April 1998. Racial distribution was: 2331 Caucasians (83.9%), 373 Blacks (13.4%) and 75 Asiatic (2.7%). PSA was normal in 2554 (91.4%), 4 to 10 in 177 (6.3%) and greater than 10 in 64 (2.3%). DRE was normal in 2419 (86.3%), suspicious in 347 (12.4%) and characteristic for cancer in 37 (1.3%). Men with abnormal DRE and/or PSA had transrectal prostate biopsy indicated. Results: 461 biopsies were done and 78 tumors was detected (prevalence = 2.8%). Prevalence was progressively higher with age (p < 0.001), PSA level (p < 0.0001) and DRE findings (p = 0.0216). Cancer prevalence in Blacks was 1.65 times higher than in Caucasians (p > 0.05) and 94.9% of detected tumors were moderately or poorly differentiated. Sensibility, specificity, positive predictive value, negative predictive value and total accuracy for PSA were respectively: 66.6%; 89.7%; 51.7%; 94.2% and 86.5%. For DRE, the respective values were: 49.1%; 79.4%; 50.9%; 78.3% and 70.3%. Conclusions: prostate cancer prevalence in the studied population (2.8%) was similar to that of other countries populations. Cancer prevalence in blacks was 1.65 times higher than in Caucasians (difference was not statistically significant). Cancer prevalence becomes higher with aging. The association of DRE and PSA is of paramount importance for cancer diagnosis. The great majority of detected tumors (94.9%) was moderately and poorly differentiated. Brazil probably needs regional studies to better characterize prostate cancer epidemiology due to population heterogeneity.

Clinical and nutritional profile of individuals with Chagas disease

Chagas disease (CD), caused by the protozoan Trypanossoma cruzi, affects approximately 18 million individuals in the Americas, 5 million of which five in Brazil. Most chronic sufferers have either the indeterminate form of the disease, without organic compromise, or the cardiac or digestive forms. Despite the importance of this disease, there is no information on the effect of nutrition on CD evolution. We evaluated the clinical-nutritional profile of individuals with CD treated at the Tropical Diseases Nutrition Out-Patient Clinic of the Botucatu School of Medicine, UNESP.A retrospective cohort study was performed between 2002 and 2006, on 66 patients with serum and parasitological diagnosis of CD. Epidemiological, clinical, nutritional, and biochemical data were collected, including gender, age, skin color, smoking, alcoholism, physical activity, weight, stature, body mass index, abdominal circumference, glycemia, and lipid profile. Fifty-three percent were mate and 47% female; 96% were white skinned. Mean age was 49.6±6.36 years. The predominant form was indeterminate in 71 %; smoking and drinking were recorded in 23% and 17%, respectively. Sedentariness predominated in 83%, and 55% presented increased abdominal circumference. Most, 94%, were overweight or obese. The biochemical exam revealed hyperglycemia in 12% and dyslipidemia in 74%. These findings suggest that the Chagas population presents co-morbidities and risk factors for developing chronic non-transmissible diseases, including cardiovascular diseases, making CD evolution even worse. © 2007 by The Brazilian Journal of Infectious Diseases and Contexto Publishing. All rights reserved.

Evaluation of clinical periodontal conditions in smokers and non-smokers

Given that tobacco smoking habit is a risk factor for periodontal diseases, the aim of this study was to compare clinical periodontal aspects between smokers and non-smokers. The clinical status were assessed in 55 patients, 29 smokers and 26 non-smokers, aged 30 to 50 years, with mean age of 40. The clinical parameters used were: probing depth (PD), plaque index (PI), gingival index (GI), clinical attachment level (CAL), gingival recession (GR) and gingival bleeding index (GBI) for arches (upper and lower ) and teeth (anterior and posterior). Tooth loss was also evaluated in both groups. Multiple regression analysis showed: tendency of greater probing depth and clinical attachment level means for smokers; greater amount of plaque in smokers in all regions; greater gingival index means for non-smokers with clinical significance (p<0.05) in all regions. Although, without statistical significance, the analysis showed greater gingival bleeding index means almost always for nonsmokers; similar gingival recession means in both groups and tendency of upper tooth loss in smokers and lower tooth loss in non-smokers. The findings of this study showed that clinical periodontal parameters may be different in smokers when compared to non-smokers and that masking of some periodontal signs can be a result of nicotine's vasoconstrictor effect.

Autoimmunity does not contribute to the highly prevalent glucose metabolism disturbances in a Japanese Brazilian population

The Japanese Brazilian population has one of the highest prevalences of diabetes worldwide. Despite being non-obese according to standard definitions, their body fat distribution is typically central. We investigated whether a subset of these subjects had autoantibodies that would suggest a slowly progressive form of type 1 diabetes. A total of 721 Japanese Brazilians (386 men) in the 30- to 60-year age group underwent clinical examination and laboratory procedures, including a 75-g oral glucose tolerance test and determinations of serum autoantibodies. Antibodies to glutamic acid decarboxylase (GADab) were determined by radioimmunoassay and to thyroglobulin (TGab) and thyroperoxidase (TPOab) by flow-cytometry assays. Mean body mass index was 25.2 ± 3.8 kg/m2, but waist circumference was elevated according to the Asian standards. Diabetes, impaired glucose tolerance, and impaired fasting glycemia were found in 31%, 22%, and 22%, respectively, and 53% of the subjects had metabolic syndrome. Glutamic acid decarboxylase (GADab) was positive in 4.72%, TGab in 9.6%, and TPOab in 10% of the whole sample. When participants were stratified according to the presence of thyroid antibodies, similar frequencies of GADab were found in positive and negative groups. The prevalence rates of glucose metabolism disturbances did not differ between GADab positive and negative groups. Our data did not support the view that autoimmune injury could contribute to the high prevalence of diabetes seen in Japanese Brazilians, and the presence of co-morbidities included in the spectrum of metabolic syndrome favors the classification as type 2 diabetes.