Intranasal Liposomal Formulation of Spike Protein Adjuvanted with CpG Protects and Boosts Heterologous Immunity of hACE2 Transgenic Mice to SARS-CoV-2 Infection.

Mucosal vaccination appears to be suitable to protect against SARS-CoV-2 infection. In this study, we tested an intranasal mucosal vaccine candidate for COVID-19 that consisted of a cationic liposome containing a trimeric SARS-CoV-2 spike protein and CpG-ODNs, a Toll-like receptor 9 agonist, as an adjuvant. In vitro and in vivo experiments indicated the absence of toxicity following the intranasal administration of this vaccine formulation. First, we found that subcutaneous or intranasal vaccination protected hACE-2 transgenic mice from infection with the wild-type (Wuhan) SARS-CoV-2 strain, as shown by weight loss and mortality indicators. However, when compared with subcutaneous administration, the intranasal route was more effective in the pulmonary clearance of the virus and induced higher neutralizing antibodies and anti-S IgA titers. In addition, the intranasal vaccination afforded protection against gamma, delta, and omicron virus variants of concern. Furthermore, the intranasal vaccine formulation was superior to intramuscular vaccination with a recombinant, replication-deficient chimpanzee adenovirus vector encoding the SARS-CoV-2 spike glycoprotein (Oxford/AstraZeneca) in terms of virus lung clearance and production of neutralizing antibodies in serum and bronchial alveolar lavage (BAL). Finally, the intranasal liposomal formulation boosted heterologous immunity induced by previous intramuscular vaccination with the Oxford/AstraZeneca vaccine, which was more robust than homologous immunity.

Impact of the COVID-19 Pandemic on Cervical Cancer Screening in São Paulo State, Brazil.

INTRODUCTION: The early identification of precursor lesions followed by appropriate treatment prevents development of cervical cancer and its consequences. OBJECTIVE: The present study evaluated the influence of the COVID-19 pandemic on cervical cancer screening by comparing the quantity of tests to detect cervical cellular changes performed in São Paulo state in 2019, prior to the detection of SARS-CoV-2 in Brazil, to the first (2020) and second (2021) years following its appearance. MATERIALS AND METHODS: Data from Fundação Oncocentro de São Paulo (FOSP), the agency that analyses approximately 220,000 Papanicolaou (Pap) tests annually, were reviewed. RESULTS: A median of 1,835 Pap tests were performed in 55 municipalities in 2019. This was reduced to 815 tests in 2020, a 56% decrease (p = 0.0026). In 2021, the median number was 1,745, a 53% increase over 2020 levels (p = 0.0233). The 26 municipalities with >1,000 tests in 2020 had a median reduction from 4,433 in 2019 to 2,580 in 2020 (p = 0. 0046). The 29 municipalities with <1,000 tests had a median reduction from 951 in 2019 to 554 in 2020 (p < 0.0001). There was a 44% reduction in the number of follow-up cytological evaluations from 2019 to 2020, followed by a 30% increase in the following year. However, the percentage of women with a normal finding or with any abnormality remained unchanged. The findings from a histological evaluation of women in São Paulo city indicated that the percent of cases positive for CIN-1 (p < 0.0410) and CIN-3 (p < 0.0012) increased in 2020 and 2021 as compared to 2019 levels. CONCLUSION: A reduction in testing for cervical cancer in the first year of the COVID-19 pandemic, accompanied by an elevated incidence of precancerous lesions in each of the first 2 years following its initiation, may portend a subsequent increased occurrence of cervical cancer in Brazil.

Moving towards a strategy to accelerate cervical cancer elimination in a high-burden city-Lessons learned from the Amazon city of Manaus, Brazil.

The World Health Organization Call to Eliminate Cervical Cancer resonates in cities like Manaus, Brazil, where the burden is among the world's highest. Manaus has offered free cytology-based screening since 1990 and HPV immunization since 2013, but the public system is constrained by many challenges and performance is not well-defined. We obtained cervical cancer prevention activities within Manaus public health records for 2019 to evaluate immunization and screening coverage, screening by region and neighborhood, and the annual Pink October screening campaign. We estimated that among girls and boys age 14-18, 85.9% and 64.9% had 1+ doses of HPV vaccine, higher than rates for age 9-13 (73.4% and 43.3%, respectively). Of the 90,209 cytology tests performed, 24.9% were outside the target age and the remaining 72,230 corresponded to 40.1% of the target population (one-third of women age 25-64). The East zone had highest screening coverage (49.1%), highest high-grade cytology rate (2.5%) and lowest estimated cancers (38.1/100,000) compared with the South zone (32.9%, 1.8% and 48.5/100,000, respectively). Largest neighborhoods had fewer per capita screening locations, resulting in lower coverage. During October, some clinics successfully achieved higher screening volumes and high-grade cytology rates (up to 15.4%). Although we found evidence of some follow-up within 10 months post-screening for 51/70 women (72.9%) with high-grade or worse cytology, only 18 had complete work-up confirmed. Manaus has successfully initiated HPV vaccination, forecasting substantial cervical cancer reductions by 2050. With concerted efforts during campaigns, some clinics improved screening coverage and reached high-risk women. Screening campaigns in community locations in high-risk neighborhoods using self-collected HPV testing can achieve widespread coverage. Simplifying triage and treatment with fewer visits closer to communities would greatly improve follow-up and program effectiveness. Achieving WHO Cervical Cancer Elimination goals in high-burden cities will require major reforms for screening and simpler follow-up and treatment.

High-Risk HPV Testing in Primary Screening for Cervical Cancer in the Public Health System, São Paulo, Brazil.

Every year there are approximately 16,000 new cases of cervical cancer in Brazil. Novel screening technologies may reduce this number by expanding the population coverage but also by improving the detection rate of precursor lesions. We aimed to evaluate human papillomaviruses (HPV)-DNA testing in the context of routine cervical cancer screening in the public health system of the city of São Paulo, Brazil. Women participating in the primary screening program were invited to enroll. Liquid-based cytology samples were collected and cytology and Hr-HPV DNA testing were performed in parallel. Cytologists were blind to HPV results. Women older than 24 years with a positive high-risk HPV test and/or cytology class ≥ ASC-US were referred to colposcopy. From December 2014 to December 2016, 16,102 women joined the study. High-risk human papillomavirus (HR HPV) DNA prevalence was 14.9%, whereas cytologic abnormalities were found in 7.2% of the women. Per protocol, 1,592 Hr-HPV+ women, in addition to 72 patients with cytologic classification > low-grade squamous intraepithelial lesion (LSIL) were referred. A total of 80 cervical intraepithelial neoplasia (CIN2+) cases were diagnosed, 79 were Hr-HPV DNA+ and 18 had normal cytology. Hr-HPV DNA detected a significant number of patients with premalignant lesions missed by cytology and all 16 CIN3+ cases were Hr-HPV DNA+ HPV genotyping may be useful in the management of Hr-HPV+ women, reducing the burden of colposcopic referral for those harboring genotypes with a weaker association to CIN3+ Use of HPV-DNA testing was shown to be feasible and advantageous over current cytologic screening in the public health system.

Self-sampling coupled to the detection of HPV 16 and 18 E6 protein: A promising option for detection of cervical malignancies in remote areas.

OBJECTIVE: To evaluate both the performance and acceptability of a method coupling self-sampling with detection of cervical malignancy via elevated HPV 16 and 18 E6 oncoproteins (OncoE6™ Cervical Test) in remote areas in Brazil. METHODS: Women living in rural villages in proximity to Coari city, Amazonas, Brazil were invited to participate in a cervical cancer screening study. 412 subjects were enrolled; there were no refusals. In addition to E6 protein detection, DNA was extracted from the brushes and evaluated for HPV genotypes by PCR (PGMY09/11), followed by typing by the Papillocheck™ if positive. Subjects who were found to be positive for OncoE6 or HPV-DNA were referred for colposcopy. RESULTS: For 110 subjects (27%) this was the first cervical cancer exam. Overall the HPV-DNA prevalence was 19.1% (n = 79); 1.4% (n = 6) were positive by the OncoE6 Test. Fifty-six women attended the invitation for colposcopy where nine had an abnormal cervix and were subsequently biopsied. Histopathological analysis revealed 2 CIN3, 2 carcinomas and 5 CIN1. OncoE6 called two out of the three HPV 16 or 18 associated CIN3+ lesions. CONCLUSIONS: The findings suggest that self-administered sample collection in combination with OncoE6 Test is feasible in this population. This could enable expanded screening coverage while ensuring a high specificity which is imperative given the remote geographic location, since women bearing abnormal test results would necessitate travel and logistical burden to access colposcopy and treatment.

Influence of Prior Knowledge of Human Papillomavirus Status on the Performance of Cytology Screening.

OBJECTIVES: This study aimed to evaluate the influence of prior knowledge of human papillomavirus (HPV) status in cervical cytopathology readings. METHODS: Participants comprised 2,376 women who underwent parallel cytology and HPV-DNA testing. Smears were read twice by the same team, first with previous knowledge of HPV-DNA status. RESULTS: Overall, 239 (10.2%) smears had their cytology classification altered by the HPV-informed review. Cytology readings with prior knowledge of the HPV status revealed 10.5% of abnormal smears (atypical squamous cells of undetermined significance or higher), while without prior knowledge, this rate dropped to 7.6%. When HPV status was informed, a significant increase in all categories of altered smears was observed. Cytology with prior knowledge of HPV status detected more cervical intraepithelial neoplasia grade 2 or higher (CIN 2+) compared with blinded: 86.7% vs 60.0%. CONCLUSIONS: Our data indicate that cytology interpreted with prior knowledge of the HPV status provides higher sensitivity for CIN 2+ lesions while marginally reducing the overall specificity compared with HPV status blinded cytology.

Multiple HPV genotype infection impact on invasive cervical cancer presentation and survival.

BACKGROUND: Invasive cervical cancer (ICC) is the third most common malignant neoplasm affecting Brazilian women. Little is known about the impact of specific HPV genotypes in the prognosis of ICC. We hypothesized that HPV genotype would impact ICC clinical presentation and survival. METHODS: Women diagnosed with ICC at the Instituto do Câncer do Estado de São Paulo (ICESP) between May 2008 and June 2012 were included in the study and were followed until December 2015. HPV genotype was detected from formalin-fixed paraffin-embedded (FFPE) tumor tissue samples using Onclarity™ system (BD Viper™ LT automated system). RESULTS: 292 patients aged 50±14 years were analyzed. HPVDNA was detected in 84% of patients. The HPV genotypes studied were: HPV16 (64%), HPV18 (10%), HPV33-58 (7%), HPV45 (5%), HPV31 (4%) and other high-risk HPV genotypes (11%). HPV genotypes showed different distributions regarding histological type and clinical stage. Patients were followed for 35±21 months. The overall survival at 5 years after diagnosis of cervical cancer was 54%. Age, clinical staging, histological type and multiple HPV genotypes infection detected in the same tumor specimen were associated with poorer overall survival on multivariate Cox proportional hazard analysis (p<0.05). No specific HPV genotype affected survival. CONCLUSION: Multiple HPV genotype infection was associated with poorer ICC survival in our study, compared with single genotype infection. HPV genotyping from FFPE tumor tissue using an automated assay such as the Onclarity BD™ assay provides a simpler alternative for routine clinical use. IMPACT: This is the largest study employing an automated HPV genotyping assay using FFPE of ICC. Multiple HPV genotype infection adversely influenced survival.

HPV genotype distribution in Brazilian women with and without cervical lesions: correlation to cytological data.

BACKGROUND: Human Papillomavirus (HPV) genotype distribution varies according to the method of assessment and population groups. This study analyzed type-specific HPV infections among women ranging from 14-95 years old, displaying normal and abnormal cytology, from São Paulo and Barretos cities, Brazil. METHODS: Women found positive for High Risk-HPVs DNA by either the Hybrid Capture 2 (HC2) or Cobas HPV Test (n = 431) plus a random sample of 223 negative by both assays and 11 samples with indeterminate results, totalizing 665 samples, were submitted to HPV detection by the PapilloCheck test. Cytological distribution included 499 women with a cytological result of Negative for Intraepithelial Lesion or Malignancy and 166 with some abnormality as follows: 54 Atypical Squamous Cells of Undetermined Significance; 66 Low-Grade Squamous Intraepithelial Lesion; 43 High-Grade Squamous Intraepithelial Lesion and 3 (0.5 %) Invasive Cervical Cancer. RESULTS: From the 323 samples (48.6 %) that had detectable HPV-DNA by the PapilloCheck assay, 31 were HPV negative by the cobas HPV and HC2 assays. Out of these 31 samples, 14 were associated with HR-HPVs types while the remaining 17 harbored exclusively low-risk HPVs. In contrast, 49 samples positive by cobas HPV and HC 2 methods tested negative by the PapilloCheck assay (19.8 %). Overall, the most frequent HR-HPV type was HPV 16 (23.2 %), followed by 56 (21.0 %), 52 (8.7 %) and 31 (7.7 %) and the most frequent LR-HPV type was HPV 42 (12.1 %) followed by 6 (6.2 %). Among the HR-HPV types, HPV 56 and 16 were the most frequent types in NILM, found in 19.1 and 17.7 % of the patients respectively while in HSIL and ICC cases, HPV 16 was the predominant type, detected in 37.2 and 66.7 % of these samples. CONCLUSIONS: In the population studied, HPV 16 and 56 were the most frequently detected HR-HPV types. HPV 56 was found mainly in LSIL and NILM suggesting a low oncogenic potential. HPV 16 continues to be the most prevalent type in high-grade lesions whereas HPV 18 was found in a low frequency both in NILM and abnormal smears. Surveillance of HPV infections by molecular methods is an important tool for the development and improvement of prevention strategies.

Critical Analyses of the Introduction of Liquid-Based Cytology in a Public Health Service of the State of São Paulo, Brazil.

OBJECTIVE: The aim of this study was to compare the performance of the current conventional Pap smear with liquid-based cytology (LBC) preparations. STUDY DESIGN: Women routinely undergoing their cytopathological and histopathological examinations at Fundação Oncocentro de São Paulo (FOSP) were recruited for LBC. Conventional smears were analyzed from women from other areas of the State of São Paulo with similar sociodemographic characteristics. RESULTS: A total of 218,594 cases were analyzed, consisting of 206,999 conventional smears and 11,595 LBC. Among the conventional smears, 3.0% were of unsatisfactory preparation; conversely, unsatisfactory LBC preparations accounted for 0.3%. The ASC-H (atypical squamous cells - cannot exclude high-grade squamous intraepithelial lesion) frequency did not demonstrate any differences between the two methods. In contrast, the incidence of ASC-US (atypical squamous cells of undetermined significance) was almost twice as frequent between LBC and conventional smears, at 2.9 versus 1.6%, respectively. An equal percentage of high-grade squamous intraepithelial lesions were observed for the two methods, but not for low-grade squamous intraepithelial lesions, which were more significantly observed in LBC preparations than in conventional smears (2.2 vs. 0.7%). The index of positivity was importantly enhanced from 3.0% (conventional smears) to 5.7% (LBC). CONCLUSIONS: LBC performed better than conventional smears, and we are truly confident that LBC can improve public health strategies aimed at reducing cervical lesions through prevention programs.

Clinical characteristics of women diagnosed with carcinoma who tested positive for cervical and anal high-risk human papillomavirus DNA and E6 RNA.

High-risk human papillomavirus (hrHPV) is an essential cause of cervical carcinoma and is also strongly related to anal cancer development. The hrHPV E6 oncoprotein plays a major role in carcinogenesis. We aimed to evaluate the frequency of hrHPV DNA and E6 oncoprotein in the anuses of women with cervical carcinoma. We analyzed 117 women with cervical cancer and 103 controls for hrHPV and the E6 oncogene. Positive test results for a cervical carcinoma included 66.7 % with hrHPV-16 and 7.7 % with hrHPV-18. One case tested positive for both HPV variants (0.9 %). The samples from the anal canal were positive for HPV-16 in 59.8 % of the cases. Simultaneous presence of HPV in the cervix and anal canal was found in 53.8 % of the cases. Regarding expression of E6 RNA, positivity for HPV-16 in the anal canal was found in 21.2 % of the cases, positivity for HPV-16 in the cervix was found in 75.0 %, and positivity for HPV-18 in the cervix was found in 1.9 %. E6 expression in both the cervix and anal canal was found in 19.2 % of the cases. In the controls, 1 % tested positive for HPV-16 and 0 % for HPV-18. Anal samples from the controls showed a hrHPV frequency of 4.9 % (only HPV16). The presence of hrHPV in the anal canal of women with cervical cancer was detected at a high frequency. We also detected E6 RNA expression in the anal canal of women with cervical cancer, suggesting that these women are at risk for anal hrHPV infection.

Cervical cytology, HPV DNA and mRNA - a comparative study in 162 patients / Citologia cervicovaginal, DNA HPV e RNAm - estudo comparativo em 162 pacientes

In contrast to the general improvement of the socioeconomic status of the Brazilian population, pathologies that are characteristic of poor health assistance persist. Among those, cervical cancer (CC) is emblematic; it still presents a persistently high incidence. Objective: to compare the performance of cervical cytology to HPV DNA and mRNA detection methods in 162 patients undergoing routine gynecological clinical practice. Methods:a total of 162 patients attended during routine gynecological examination in a private clinic in Florianópolis, Santa Catarina, Brazil, had cervical samplescollected and processed for cytopathological and molecular tests, conventional PCR and NASBA. Positive samples positive for HPV DNA were submittedto Type-Specific PCR (TS-HPV PCR). Patients with altered smears were submitted to colposcopy and biopsy. Results: among the 162 samples, 19.8%(32/162) had altered smears, being 4/32 classified as ASC-H, 9/32 as ASC-US, 9/32 as LSIL and 10/32 as HSIL. Biopsies revealed nine cases of CIN I,nine CIN II and one CIN III, while seven were negative for cervical neoplasia. Overall, HPV DNA was detected in 38.3% (62/162) of the samples and HPV E6/E7 mRNA expression was found in 13.6% (22/162). Using TS-HPV PCR, HPV 16 was the most frequent type, found in 8% of the samples (5/62).Considering CIN2+ the gold-standard, cytology had 38.5% of specificity. Sensitivity and specificity of HPV-DNA PCR and NASBA were, respectively,100% and 60%; 18.7% and 68.7%. Conclusion: mRNA E6/E7 expression was not a highly specific or sensitive marker for prevalent cervical disease while HPV DNA may be used for cervical cancer screening only in conjunction to more specific adjuvant tests.

Em contraste com a melhora geral da situação socioeconômica da população brasileira, patologias que são características de uma deficiente assistência à saúde persistem. Entre elas, o câncer cervical (CC) é emblemático, ainda apresentando uma persistente alta incidência. Objetivo: avaliaro desempenho da citologia e de métodos de detecção de DNA e RNAm de HPV em 162 pacientes submetidas a prática clínica ginecológica de rotina.Métodos: cento e sessenta e duas pacientes atendidas em uma clínica particular de Florianópolis, Santa Catarina, Brasil, tiveram amostras cervicais coletadas e processadas para estudo citopatológico e molecular; PCR convencional e NASBA. Amostras positivas para o DNA do HPV foram submetidas àPCR tipo-específica (PCR HPV-TE). Resultados: entre as 162 amostras, 19,8% (32/162) apresentaram esfregaços alterados, sendo 4/32 classificadas comoASC-H, 9/32 como ASC-US, 9/32 como LSIL e 10/32 como HSIL. Biópsias revelaram nove casos de NIC I, nove casos de NIC II e um caso de NIC III. ODNA do HPV foi detectado em 38,3% (62/162) das amostras. Expressão de E6/E7 (RNAm) foi encontrada em 13,6% (22/162) das amostras. Utilizando a PCR tipo-específica (HPV-TE), o HPV 16 foi o tipo mais frequente, encontrado em 8% (5/62) das amostras HPV+. Considerando NIC 2+ o padrão-ouro,a especificidade da citologia foi de apenas 38,5%, enquanto a sensibilidade e a especificidade da PCR DNA e RNAm foram, respectivamente, 100% e 60%;18,7% e 68,7%. Conclusão: a expressão de E6/E7 RNAm não se mostrou um marcador altamente específico ou sensível para doença cervical prevalente,enquanto o DNA HPV pode ser utilizado para rastreamento apenas em conjunto com testes adjuvantes mais específicos.

Performance of RT-PCR in the detection of Streptococcus agalactiae in the anogenital tract of pregnant women.

INTRODUCTION: Infection with Group B Streptococcus (GBS) is the most frequent in the first weeks of life of a newborn. The identification of pregnant women with GBS colonization may reduce neonatal infection. METHODS: This cross-sectional study evaluated the performance of real-time polymerase chain reaction (RT-PCR) to detect GBS colonization in the anogenital tract of pregnant women. Anogenital swabs were collected from 266 pregnant women from December 2010 to August 2011. GBS was detected using culture (gold standard) and RT-PCR to determine sip gene expression. The presence of DNA was confirmed using betaglobin amplification, and the guanidine technique was used for DNA extraction. When results were discordant, the test was repeated using conventional PCR. The results were evaluated to determine sensitivity, specificity, positive and negative predictive values and accuracy. RESULTS: Of the 266 samples collected, 254 were adequate for analysis. Prevalence was 28.7% using the gold standard criterion and 38.2% using RT-PCR. The comparison of RT-PCR with culture revealed a sensitivity of 89% (95% CI 0.81-0.96), specificity of 82% (95% CI 0.76-0.87), positive predictive value of 67% (95% CI 0.57-0.76) and negative predictive value of 94% (95% CI 0.91-0.99). CONCLUSION: Further studies using other DNA extraction techniques, targeting other GBS genes and using sample enhancement before RT-PCR should be conducted to determine whether the sensitivity and specificity recommended by the CDC may be reached using the same thermal cycler.

Prevenção do câncer cervical: associação da citologia oncótica a novas técnicas de biologia molecular na detecção do papilomavírus humano (HPV) / Cervical cancer prevention: association of new techniques of molecular biology with the oncotic cytology in detection of human papillomavirus (HPV)

O câncer do colo uterino é conhecido como uma das causas mais frequentes de óbito na população feminina em todo o mundo. A infecção persistente por papilomavírus humano (HPV) é o principal fator de risco para o câncer cervical e suas lesões precursoras. Objetivo: avaliar os métodos diagnósticos do câncer cervical: citologia oncótica, DNA do HPV pela PCR, detecção do RNAm das proteínas E6 e E7 dos HPVs de alto risco, para acrescentar novos marcadores na detecção do HPV em amostras com citologia alterada. Métodos: foram avaliados os resultados da citologia oncótica de3.000 pacientes, atendidas no Laboratório Vital, Nonoai - RS, de março de 2009 a janeiro de 2010, classificando-as conforme a faixa etária e codificação de Bethesda, 2001. Foram selecionados dois grupos: o grupo A incluiu pacientes com citologia prévia alterada e o grupo B, pacientes com citologia sem anormalidades. Foram coletadas 47 amostras, 22 do grupo A e 25 do grupo B. Três amostras do grupo B apresentaram inibidores da PCR e foram excluídas.A análise dos dados considerou 22 amostras em cada grupo. A detecção do HPV foi feita pela reação em cadeia da polimerase (PCR) utilizando-se os primers consenso MY9/11 e GP5+/6+. O RNAm das proteínas E6 e E7 foi detectado pelo método Real-Time NASBA. Resultados: nas amostras do grupo A, houve detecção do DNA/HPV pela PCR em 14 (64%) amostras, e RNAm das proteínas E6 e E7 do HPV em seis (27%) amostras. Todas as 22 amostras do grupoB foram negativas nos métodos moleculares. Conclusão: neste estudo, a citologia prévia alterada associada a testes moleculares indica a necessidade de monitoramento ou intervenção terapêutica das pacientes. Assim, conclui-se que a citologia oncótica associada à detecção do DNA do HPV pela PCR e à expressão das oncoproteínas E6 e E7 é a alternativa para identificar precocemente pacientes com elevado risco de desenvolver o câncer cervical.

Worldwilde, the cervical cancer is known as one of the most frequent cause of death in female population. Persistent infection with oncogenic human papillomavirus (HPV) is the main risk factor for cervical cancer and its precursor lesions. Objective: to evaluate the diagnostic methods of cervical cancer: oncotic cytology, HPV DNA by PCR, detection of mRNA E6 / E7 proteins of high-risk HPV, to add new markers in the detection of HPV.Methods: were evaluated 3,000 results of oncotic cytology from patients referred to Vital Laboratory, Nonoai- RS, Brazil, from March 2009 to January2010. The results were grouped according age and Bethesda classification, 2001. Group A included patients with previous altered cytology and GroupB patients without abnormalities in cytology. To molecular tests were collected 47 samples, 22 from group A and 25 from group B. Three samples from group B presented PCR inhibitors and were excluded. The detection of HPV was made by polymerase chain reaction (PCR) using the consensus primers MY9/11 and GP5 +/6 +. The mRNA E6 / E7 proteins were detected by real-time NASBA. Results: in samples of group A, DNA/HPV was detected in 14(64%) samples, and mRNA E6 / E7 proteins of HPV was detected in 6 (27%) samples. All 22 samples of group B were negative to molecular methods.Conclusion: in this study, the previous altered cytology associated with molecular methods indicates the need for monitoring or therapeutic intervention of patients. The oncotic cytology associated with the detection of HPV DNA by PCR and the expression of oncoproteins E6 / E7 are alternatives to identify early lesions in patients with high risk of developing cervical cancer.

Cistatina C: um novo marcador para a filtração glomerular comparada ao clearance de creatinina e a creatinina sérica / Cystatin C: a new marker for glomerular filtration rate compared to creatinine clearance and serum creatinine

A Cistatina C é uma proteína de baixo peso molecular (- 13 kDa) produzida constantemente em todas as células nucleadas. Esta molécula é livremente filtrada no glomérulo renal, reabsorvida e catabolizada no túbulo proximal, sendo os níveis séricos dependentes e indicadores da função de filtração glomerular. Esta revisão aborda o desempenho da Cistatina C em relação aos testes usualmente empregados na rotina laboratorial para avaliação da função renal. Verifica-se que a Cistatina C é um marcador confiável da filtração glomerular mais sensível e específico que as determinações de creatinina sérica e clearance de creatinina, e pode ser uma alternativa atrativa, especialmente quando a população pediátrica é considerada.