Avaliação clínica e microbiológica das condições de saúde oral dos receptores de transplante renal do Hospital Universitário Onofre Lopes, Natal RN

Introduction: The chronic immunosuppression promotes the development of changes in the oral cavity of the kidney transplant recipients (KTR), however with the exception of gingival overgrowth, little is known regarding the prevalence of oral lesions in this population. Objective: To evaluate the prevalence of clinical and microbiological conditions of the oral cavity of the recipients of kidney transplantation and the associated factors. Methods: This was a cross-sectional study which examined 96 clinical KTR and experimental where collected saliva stimulated them to identify Candida sp. Data collection consisted of evaluation stomatologic, socio-demographic, clinical aspects of transplantation, condition of oral hygiene and dental caries, yonder to questions about knowledge of oral alteration after transplantation. Results: Of the total, 66.7% of KTR had some type of oral manifestation. The most common was saburral tongue, followed by gingival overgrowth, with both oral manifestations related to gender and concomitant use of cyclosporine and nifedipine (p <0.05). Tacrolimus showed a protective effect for gingival overgrowth (OR = 0.13). The oral hygiene was associated with saburral tongue(p = 0.03) and severity of gingival overgrowth (p = 0.0001). Oral candidiasis was diagnosed in 17.7% of patients and Candida albicans was isolated most frequently in the saliva of RTR with a colonization of 58.3%. The average DMF-T increased with age. The method of oral hygiene was most used brush and toothpaste to 61.5%. Changes in the oral cavity was seen in 54.2% of KTR, citing as the main growth and gingival ulcers. Instructions for oral hygiene after transplantation were neglected for 61.5% of RTR. Dry mouth and halitosis were reported in 30.2% and 36.5% respectively. Conclusions: More the half of the KTR had at least one injury of the mouth, the immunosuppressive drugs and oral hygiene are associated with these alterations. Prospective cohort studies are needed to elucidate the relationship between oral manifestations and levels of drug and risk of oral manifestations occur over time. The kidney transplant recipients showed to be aware of oral alterations occurred after transplantation and uninformed about the oral hygiene instructions. With regard to hygiene, the incidence of caries was considered high, conditions of risk were identified and improvements in primary attention should be encouraged and reflected in the monitoring of renal transplant

Identificação e fatores de virulência de Candida spp isoladas da cavidade bucal de transplantados renais do hospital universitário Onofre Lopes em Natal-RN

Despite Candida species are often human commensals isolated from various oral sites such as: tongue, cheek and palatal mucosa plus subgingival region, there are some properties linked to the organism commonly known as virulence factors which confer them the ability to produce disease. Oral candidiasis is one of the main oral manifestations reported in literature related to kidney transplant patients. The objectives of the present study were to identify and investigate virulence factors of yeasts isolated from the oral cavity of kidney transplant recipients admitted at the Hospital Universitário Onofre Lopes, in Natal RN. Seventy Candida species isolated from 111 kidney transplant recipients were investigated in this study. Identification of the isolates was performed by using the evidence of germ tube formation, hypertonic broth, tolerance to grow at 42°C, micromorphology and biochemical profiles. We observed a high rate of isolation of yeasts from the oral cavity of kidney transplant recipients (63.1%) being C. albicans was the most prevalent species. Oral candidiasis was diagnosed in 14.4% of transplant recipients. We evaluated virulence properties of the isolates regarding to: biofilm formation on polystyrene microplates as well as XTT reduction, adherence to acrylic resin and human buccal epithelial cells and proteinase activity. Most isolates were able to form biofilm by the method of adhesion to polystyrene. All isolates of Candida spp. remained viable during biofilm formation when analyzed by the method of XTT reduction. The number of CFU attached to the acrylic resin suggested high adherence for C. parapsilosis. C. albicans isolates showed higher median adherence to human buccal epithelial cells than non-C. albicans Candida isolates. Nevertheless, this difference was not statistically significant. C. dubliniensis showed low ability to adhere to plastic and epithelial cells and biofilm formation. Proteolytic activity was observed for all the isolates investigated, including the unique isolate of C. dubliniensis. There was a statistically significant association between proteinase production and the presence of oral candidiasis. Studies related to oral candidiasis in renal transplant recipients are limited to clinical and epidemiological data, but investigations concerning Candida spp. virulence factor for this group of individuals are still scarce. We emphasize the importance of studies related to virulence factors of yeasts isolated from this population to contribute to the knowledge of microbiological aspects of oral candidiasis

Características genotípicas e fenotípicas de Candida Albicans isoladas da cavidade bucal de pacientes transplantados renais com ênfase na ação do extrato bruto de Eugenia uniflora em fatores de virulência

Candida albicans is a diploid yeast that in some circumstances may cause oral or oropharyngeal infections. The investigation of natural products is mandatory for the discovery of new targets for antifungal drugs development. This study aimed to determine the genotypes of 48 clinical isolates of C. albicans obtained from the oral cavity of kidney transplant patients from two distinct geographic regions of Brazil. In addition, we investigated three virulence factors in vitro: phospholipase activity, morphogenesis and the ability to evade from polymorphonuclear neutrophils. The expression of these virulence factors in vitro was also investigated in the presence of the crude extract of Eugenia uniflora. The genotype A was the most prevalent (30 isolates; 62.5%), followed by genotype C (15 isolates; 31.5%) and genotype B (3 isolates; 6.25%). When microsatellite technique with primer M13 was applied, 80% of the isolates from the South were placed within the same cluster. All Genotype C strains were grouped together within two different clusters. Genotype C was considered more resistant to PMNs attack than genotypes A and B. Strains isolated from the South of Brazil showed higher ability to combat PMNs phagocytosis. We found a high rate of genotype C strains isolated from the oral cavity of this group of patients. The crude extract of E. uniflora inhibited proper hypha formation and phagocytosis by PMNs, but had no significant effect on phospholipase activity. This study characterized oral C. albicans strains isolated from kidney transplant recipients and will contribute for the better understanding of the pathogenesis and alternative therapeutics for oral candidiasis

Análise quantitativa das células CD4 e sua importância no diagnóstico de lesões orais em pacientes HIV positivos

The oral manifestations due to HIV infection are, a lot of times, the first clinical signs of the disease. These injuries may also function as beepers and sentries of the curse and progression of the HIV infection and AIDS. The objective of this work was to evaluate the prevalence of the oral injuries in HIV positive patients, relating them with the CD4+ cells counting and the viral load in patients from the Hospital of Infected contagious Gizelda Trigueiro in Natal-RN. One hundred and one patients were evaluated, where after the clinical exam of the oral cavity, these ones were conducted to the peripheral blood collection for the counting of CD4+ lymphocytes. We observed a prevalence of 25,6%, that is, 31 cases. The Oral Candidiasis was the most commum injure, followed by Oral Hairy Leukoplakia, linear gingival erytema, lips herpes, gingivitis and periodontitis - HIV. The average counting of cells CD4+ of the injury carrying patients was of 250 cells/mm3. We did not observe relation between the presence of injuries and the viral load of the individuals

Triagem antifúngica de extratos obtidos de espécies vegetais do nordeste brasileiro

The increase in the incidence of fungal infections due to the drug-resistance or to the number of patients with immune alterations such as AIDS, chemotherapy or organ transplantation, has done the research necesseray for new antifungal drugs. The species from Northeastern Brazil may become an important source of innovative natural molecules. To evaluate the antifungal activity of 10 medicinal plants from Northeastern Brazil, traditionally used as antimicrobial agents, 30 crude extracts (CE) were tested in vitro against four standard species of Candida spp. The CE most promising of these plants were evaluated against yeasts of the oral cavity of kidney transplant patients and through a bioassay-guided fractionation. The extracts form leaves of E. uniflora, the stem bark of L. ferrea and leaves of P. guajava showed significant activity against all yeasts evaluated, with MIC values between 15.62 and 62.5 μg/mL. E. uniflora also showed fungicidal properties against all yeasts, especially against Candida dubliniensis. In patients with immune systems compromised, such as transplanted, oral candidiasis manifests mainly due to immunosuppressive therapy, and resistance to conventional antifungals. The CE of E. uniflora presented range of MIC values between 1.95 to 1000 μg/mL, and lower MIC50 and MIC90 values were observed against C. non-albicans. Due the better results, the CE of E. uniflora was elected to performe the bioassay-guided fractionation. Thus it was possible to obtain enriched fractions, which showed good inhibitory ability against ATCC strains of Candida spp. It was also possible to perform experiments to verify the production of biofilm in two strains of C. dubliniensis and action of extracts and fractions on the same. With this, we observed a behavior between the yeast ATCC and clinical isolate. In addition, CE, fractions and subfractions of E. uniflora inhibit planktonic cells to preventing the growth of biofilm. The preliminary chemical characterization of the fractions obtained revealed the presence of polyphenols (especially flavonoids and tannins). Finally, the results suggests that among the plant species studied, E. uniflora showed a pattern very promising as regards the antifungal, requiring further study of purification and structural elucidation of compounds in order to verify that the antifungal effect found can be attributed to a specific compound or some mechanism depends on synergistic the mixture of polyphenols

Capacidade de adesão e formação de biofilme de Candida ssp. isoladas da cavidade oral de pacientes transplantados renais na presença do extrato de Eugenia uniflora

Candidiasis is a major oral manifestation in kidney transplant patients. Candida spp. possess essential virulence factors which contribute for the infectious process, including the ability to adhere to epithelial cells and biofilm formation. The extract obtained from the leaves of Eugenia uniflora [acetone: water (7:3, v/v)] has demonstrated antifungal activity against Candida spp. This study evaluated the influence of the extract of E. uniflora in adhesion to human buccal epithelial cells (HBEC) and biofilm formation of 42 strains of Candida spp. isolated from the oral cavity of kidney transplant patients. Candida spp. strains belonging to a culture collection were reactivated and phenotypically re-identified by classical and molecular methods (genotyping ABC and RAPD), when necessary, to complete the identification to the species level. For the virulence tests evaluated in vitro, yeasts were grown in the presence and absence of 1000 g/mL of the extract. A ratio of 10: 1 (Candida spp. cells x HBECs) was incubated for 1 hour at 37 ° C, 200 rpm, fixed with 10% formalin and the number of Candida cells adhered to 150 HBEC determined by optical microscope. Biofilms were formed on polystyrene microplates in the presence or absence of the extract. The quantification was performed with crystal violet staining at 570 nm. All isolates were viable and exhibited phenotypic characteristics suggestive of each species identified. Two strains presumptively identified as Candida dubliniensis belonged to this species as determined with genotyping ABC, while strains identified as belonging to the Candida parapsilosis species complex were differentiated by RAPD genotyping. Candida albicans was found to be the most adherent species to the buccal epithelia, while C. tropicalis showed remarkable biofilm formation.We could detect that the extract of E. uniflora was able to reduce adhesion to HBEC for both Candida albicans and non-Candida albicans Candida species. On the other hand, only 16 Candida spp. strains (36 %) showed reduced biofilm formation. However, two highly biofilm producer strains of C. tropicalis had an expressive reduction in biofilm formation. This study reinforces the idea that besides growth inhibition, E. uniflora may interfere with the expression of some virulence factors of Candida spp., and may be possibly applied in the future as a novel antifungal agent.