Luciane Noal Calil

p16INK4a and Ki67 expression in normal, dysplastic and neoplastic uterine cervical epithelium and human papillomavirus (HPV) infection

Cellular cycle proteins like the p16(INK4a) and the Ki67 proliferation nuclear antigen have been used as oncogenicity cellular markers. The E6 and E7 oncoproteins interact with tumor suppressor genes p53 and pRb, culminating with the p16(INK4a) overexpression. The objective of this study was to evaluate the presence of HPV-DNA in 174 cervical biopsies and correlate the different histological grades with the p16(INK4a) and Ki67 immunohistochemical expression (IHC). A cross-sectional study that enrolled a total of 174 women who underwent uterine cervical biopsies between February 2003 and December 2006, in southern Brazil, was performed. Cervical smear samples were analyzed for the presence of HPV-DNA through polymerase chain reaction (PCR), and biopsy samples were examined for p16(INK4A) and Ki67 expression through IHC techniques. The presence of HPV-DNA was observed in 89% of the tested patients, among which 52% were positive for high-risk (HR) viral types [16, 18 and 31]. Regarding p16(INK4a), an expression of 69% was observed, being expressed in 100% of the high-grade squamous lesions (HSIL) and HR-HPV-DNA positives. Ki67 expression was associated with the lesion grade, being more expressive in the most severe lesions (p<0.001). p16(INK4A) and Ki67 markers coexpression was present in 86% of the samples (p<0.001), being 100% among those positive to HR-HPV-DNA with HSIL (p<0.001). The results suggest an association between the presence of HR-HPV infection and the p16(INK4a) and Ki67 expression and which is even stronger among women with HSIL.

Chlamydia trachomatis and human papillomavirus coinfection: association with p16INK4a and Ki67 expression in biopsies of patients with pre-neoplastic and neoplastic lesions

The objective of this study was to identify the frequency of coinfection by human papillomavirus (HPV) and Chlamydia trachomatis (CT) in cervical lesions and relate it with immunohistochemical expression of p16INK4a and Ki67, both oncogenicity markers. A cross-sectional study with 86 women from primary care units in southern Brazil was conducted. Cervical swabs were collected for HPV-DNA and CT-DNA detection, through the polymerase chain reaction technique (PCR). The immunohistochemical analysis was performed on biopsy cervical tissue material to identify the expression of p16INK4a and Ki67 cell cycle markers. About 83 % were positive for HPV-DNA and 19% had coinfection with CT-DNA. Among coinfected women, 56% expressed p16INK4a. There was a statistically significant association between the histological grade of the lesion and Ki67 expression. All high-grade lesions, 50% of low-grade lesions and 31% of negative biopsies expressed Ki67 (p = 0.004). A total of 37% of coinfected women expressed both markers. In conclusion, although more than half of the coinfected patients have expressed p16INK4a and more than one third have expressed both markers, these results suggest no association between those variables. However, other studies involving larger samples are necessary to corroborate such findings.

Association of human papillomavirus and Chlamydia trachomatis with intraepithelial alterations in cervix samples

The influence of different infectious agents and their association with human papillomavirus (HPV) in cervical carcinogenesis have not been completely elucidated. This study describes the association between cytological changes in cervical epithelium and the detection of the most relevant aetiological agents of sexually transmitted diseases. Samples collected from 169 patients were evaluated by conventional cytology followed by molecular analysis to detect HPV DNA, Chlamydia trachomatis, herpes simplex virus 1 and 2,Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, andTreponema pallidum, besides genotyping for most common high-risk HPV. An association between cytological lesions and different behavioural habits such as smoking and sedentariness was observed. Intraepithelial lesions were also associated with HPV and C. trachomatis detection. An association was also found between both simple and multiple genotype infection and cytological changes. The investigation of HPV and C. trachomatisproved its importance and may be considered in the future for including in screening programs, since these factors are linked to the early diagnosis of patients with precursor lesions of cervical cancer.

LEVANTAMENTO EPIDEMIOLÓGICO DA CANDIDÍASE VULVOVAGINAL EM LAUDOS CITOPATOLÓGICOS REALIZADOS DURANTE SEIS ANOS NO ESTADO DO RIO GRANDE DO SUL, BRASIL

Vulvovaginal candidiasis (VVC) is a common vaginal infectious disease. The constant increase in the incidence of the disease may be associated with factors such as age, HIV infection, diabetes, use of hormonal contraceptive methods and cytopathological alterations. The aim of the study was to survey the prevalence of Candida sp. in cytopathology reports in the state of Rio Grande do Sul, Brazil, in the period of 2005-2010. In this observational and retrospective study a total of 121,328 cytopathology reports (CR) of cytopathology were assessed in a period of 6 years. The mean age of patients was 35 years old. Of this total, 8,582 CR (7.1%) were positive for Candida sp. More than half of patients (53%) were using oral contraceptive. Alterations in the cervix were present in 49% of the cases. The continuity of epidemiological research is needed to monitor trends over the years to better understand the factors that predispose to VVC in Brazil.

Prevalence of Superficial and Cutaneous Mycoses in Patients Attending a University Extension Activity

Docente do Departamento de Análises da Faculdade de Farmácia da Universidade Federal do Rio Grande do Sul – UFRGS. Doutor em Microbiologia ênfase em Micologia e Pós doutor em tecnologias da informática aplicada a educação. Porto Alegre. Rio Grande do Sul. Brasil. Acadêmica do curso de Farmácia da Universidade Federal do Rio Grande do Sul – UFRGS. Porto Alegre. Rio Grande do Sul. Brasil. Farmacêutica, técnica em Laboratório no Departamento de Análises da Faculdade de Farmácia da Universidade Federal do Rio Grande do Sul – UFRGS. Porto Alegre. Rio Grande do Sul. Brasil. Docente do Departamento de Análises da Faculdade de Farmácia da Universidade Federal do Rio Grande do Sul – UFRGS. Doutor em Hematologia ênfase em Citologia. Porto Alegre. Rio Grande do Sul. Brasil. 1

Differentiation of Candida albicans, Candida glabrata, and Candida krusei by FT-IR and chemometrics by CHROMagar™ Candida

Pathogenic Candida species are detected in clinical infections. CHROMagar™ is a phenotypical method used to identify Candida species, although it has limitations, which indicates the need for more sensitive and specific techniques. Infrared Spectroscopy (FT-IR) is an analytical vibrational technique used to identify patterns of metabolic fingerprint of biological matrixes, particularly whole microbial cell systems as Candida sp. in association of classificatory chemometrics algorithms. On the other hand, Soft Independent Modeling by Class Analogy (SIMCA) is one of the typical algorithms still little employed in microbiological classification. This study demonstrates the applicability of the FT-IR-technique by specular reflectance associated with SIMCA to discriminate Candida species isolated from vaginal discharges and grown on CHROMagar™. The differences in spectra of C. albicans, C. glabrata and C. krusei were suitable for use in the discrimination of these species, which was observed by PCA. Then, a SIMCA model was constructed with standard samples of three species and using the spectral region of 1792-1561cm-1. All samples (n=48) were properly classified based on the chromogenic method using CHROMagar™ Candida. In total, 93.4% (n=45) of the samples were correctly and unambiguously classified (Class I). Two samples of C. albicans were classified correctly, though these could have been C. glabrata (Class II). Also, one C. glabrata sample could have been classified as C. krusei (Class II). Concerning these three samples, one triplicate of each was included in Class II and two in Class I. Therefore, FT-IR associated with SIMCA can be used to identify samples of C. albicans, C. glabrata, and C. krusei grown in CHROMagar™ Candida aiming to improve clinical applications of this technique.

Evaluation of semi-automated cells counting in peritoneal fluid

Introduction:Currently, the cytological analysis of biological fluids, such as peritoneal fluid, is performed by manually cells counting in Fuchs-Rosenthal chamber. However, this method has a number of limitations. Because of these limitations, automatic counters have been evaluated for cell counting in this type of sample in order to make it faster and more reliable test.Objective:The aim of this study is to compare the manual and semi-automated leukocytes and erythrocytes counting in peritoneal fluid.Materials and methods:The samples were analyzed manually and using the CountessTM(Invitrogen).Results:The results showed that although there is a correlation between the two counting methods, the correlation is relatively low, for both leukocytes and erythrocytes analysis.Conclusion:The results suggest that peritoneal fluid should continue to be analyzed in Fuchs-Rosenthal chamber. However, further studies should be conducted with a greater number of samples to investigate the possibility of using automated cells counting in serous fluids and, thus, provide greater speed and quality of results.