Greater expression of the human leukocyte antigen-G (HLA-G) and interleukin-17 (IL-17) in cervical intraepithelial neoplasia: analytical cross-sectional study / Aumento da expressão do antígeno leucocitário humano-G (HLA-G) e interleucina-17 (IL-17) em neoplasia intraepitelial cervical: estudo transversal analítico

CONTEXT AND OBJECTIVE:Impaired local cell immunity seems to contribute towards the pathogenesis and progression of cervical intraepithelial neoplasia (CIN), but the underlying molecular mechanisms promoting its progression remain unclear. Identification of new molecular markers for prognosis and diagnosis of early-stage CIN may aid in decreasing the numbers of CIN cases. Several novel immunoregulatory molecules have been discovered over the past few years, including the human leukocyte antigen G (HLA-G), which through interaction with its receptors exerts important tolerogenic functions. Several lines of evidence suggest that T-helper interleukin-17 (IL-17)-producing cells (Th17 cells) may play a role in antitumor immunity. However, recent reports have implicated Th17 cells and their cytokines in both pro and anti-tumorigenic processes. The aim of the study was to evaluate the roles of HLA-G and Th17 in the immunopathogenesis of CIN I.DESIGN AND SETTING:Analytical cross-sectional study with a control group using 58 cervical specimens from the files of a public university hospital providing tertiary-level care.METHODS:We examined HLA-G and IL-17 expression in the cervical microenvironment by means of immunohistochemistry, and correlated these findings with clinical and pathological features.RESULTS:There was a greater tendency towards HLA-G and IL-17 expression in specimens that showed CIN I, thus suggesting that these molecules have a contribution towards cervical progression.CONCLUSION:These findings suggest that HLA-G and IL-17 expression may be an early marker for assessing the progression of cervical lesions.

CONTEXTO E OBJETIVO:A deficiência na imunidade celular localizada parece contribuir para a patogênese e progressão das neoplasias intraepiteliais cervicais (NIC), no entanto, ainda não está totalmente esclarecido o mecanismo molecular fundamental nesse processo de progressão. A identificação de novos marcadores moleculares de prognóstico e diagnóstico das NIC em estágios precoces pode ajudar a diminuir a quantidade de casos de NIC. Várias novas moléculas com função imunorregulatória foram descobertas nos últimos anos, inclusive o antígeno leucocitário humano G (HLA-G), que, através de interação com os receptores, tem importantes funções tolerogênicas. Diversas linhas de evidência sugerem que as células T-ajudantes produtoras de interleucina-17 (IL-17, células Th17), podem desempenhar um papel na imunidade antitumoral. Porém, recentes relatos implicaram as células Th17 e suas citocinas tanto em processos pro- quanto anti-tumorigênicos. O objetivo do estudo foi avaliar o papel do HLA-G e Th17 na imunopatogênese das NIC I.TIPO DE ESTUDO E LOCAL:Estudo transversal analítico com grupo controle em 58 espécimes cervicais dos arquivos de um hospital universitário público com assistência prestada no nível terciário.MÉTODOS:Avaliamos a expressão de HLA-G e IL-17 por imunoistoquímica no microambiente cervical, associando esses achados com as características clínico-patológicas.RESULTADOS:Houve tendência aumentada da expressão de HLA-G e IL-17 em espécimes que apresentaram NIC I, sugerindo que essas moléculas têm contribuição na progressão cervical.CONCLUSÃO:Estes resultados sugerem que a expressão do HLA-G e da IL-17 pode ser um marcador precoce para avaliar a progressão das lesões cervicais.

Métodos de indução do trabalho de parto / Methods of induction of labour

Os métodos de indução do parto podem ser divididos em estímulos naturais, estímulos exógenos diretos ou mecânicos e estímulos exógenos indiretos ou farmacológicos, cada qual apresenta suas particularidades nas indicações e contraindicações. O objetivo deste artigo foi realizar uma revisão da literatura consultando Medline/Pubmed e a Biblioteca Cochrane para avaliar a eficácia e segurança na utilização dos principais métodos de indução do trabalho de parto. Apurou-se não haver método ideal de indução do trabalho de parto. Os estímulos naturais e os métodos alternativos carecem de maiores estudos para incentivo de seu uso rotineiro. As prostaglandinas, em destaque o misoprostol, está indicada no Índice de Bishop desfavorável e a ocitocina em condições cervicais favoráveis. Os avanços no campo da biologia molecular tem corroborado que o método ideal deve atuar em sincronismo com a contratilidade uterina e a maturação cervical.(AU)

Methods of labor induction can be classified as natural stimuli, direct exogenous stimuli or mechanical and indirect exogenous stimuli or pharmacological. Which one has its peculiarities in relation to indications and contraindications. The objective of this article was to assess the efficacy and safety of the main methods of induction of labor trough the analysis of the medical literature in Medline/Pubmed and the Cochrane Library to. No ideal method of inducing labor was found. Further studies are required to encourage natural stimuli and alternative methods more often. According to Bishop scores, prostaglandins, (especially misoprostol) are unfavorable and oxytocin in case of favorable cervical environment. Advances in the field of molecular biology have confirmed that the ideal method should work simultaneously with uterine contraction and cervical ripening.(AU)

Detección de virus papiloma humano (HPV) a partir de muestras celulares de cuello uterino en base líquida: Correlación con la inmunorreactividad de la proteína p16INK4a / Detection of human papilloma virus (HPV) in liquid-based cervical samples: Correlation with protein p16INK4a expression

La citología del cuello uterino en base líquida mejora la calidad de la muestra y el material residual podría ser utilizado para realizar pruebas complementarias, como la detección del virus papiloma humano (HPV) y estudio inmunocitoquímico de biomarcadores. El propósito de este estudio fue correlacionar la presencia de HPV y la inmunoexpresión de p16INK4a en las muestras citológicas en base líquida para examinar la utilidad de estas nuevas herramientas en la detección de cáncer de cuello uterino. Las pacientes incluidas (n=67) presentaban una citología anormal o patología cervical previa. La detección y genotipificación de HPV se realizó con PCR-SPF10/LiPA (INNOLiPA Extra Amp) y para la inmunodetección de p16INK4a se utilizó el anticuerpo clon E6H4. La citología convencional proporcionó los mismos hallazgos citológicos que la citología en base líquida. La prevalencia general del HPV fue de 43,3% (29/67). El HPV16 fue el tipo viral mas frecuente (31,03%) y el 48,3% de los casos presentó infección múltiple. En el 35,8% de las muestras cervicales se detectó inmunoexpresión de la p16INK4a y ésta fue significativamente (p<0,020) asociada a la presencia de HPV. Estos resultados apoyan la evidencia que la implementación de nuevas tecnologías en la rutina diaria del laboratorio contribuye significativamente en la detección precoz del cáncer de cuello uterino y en el aporte de datos importantes para facilitar en el manejo clínico adecuado de la paciente. La detección de HPV combinada con la p16INK4a podría ser utilizado en la evaluación de pacientes con mayor riesgo a desarrollar lesiones cervicales significativas.

The liquid-based cervical cytology improves the quality of the sample and the residual sample could be used efficiently to carry out complementary tests, such as the detection of HPV DNA and the immunocytochemical biomarkers study. The purpose of this study was to correlate the presence of HPV and immunoexpression of p16INK4a in liquid-based cervical samples to examine the utility of these new tools in the detection of cervical cancer. The included patients (n=67) presented an abnormal cytology or previous cervical pathology. The HPV detection and genotyping were carried out with PCR-SPF10/LiPA (INNOLiPA Extra Amp) and for p16INK4a immunodetection was used antibody clone E6H4. The conventional cytology provided the same cytologic interpretations that those of liquid-based cytology. The overall HPV prevalence was 43.3% (29/67). HPV16 was the most frequent viral type (31.03%) and 48.3% of the cases were infected with multiple HPV types. p16INK4a immunoexpression was observed in 35.8% of liquid-based cytological samples and this was significantly (p < 0.020) associated to the HPV presence. These results support the evidence that the implementation of new technologies in the daily routine of the laboratory, contribute significantly in the early detection of cervical cancer and provide important data to help in the patient’s efficient management. The combined use of HPV detection and p16INK4a expression could be used for evaluation of patients with more risk to develop significant cervical lesions.

Identification of miR-23a as a novel microRNA normalizer for relative quantification in human uterine cervical tissues

Quantitative real-time RT-PCR (RT-qPCR) is being widely used in microRNA expression research. However, few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in microRNA RT-qPCR studies. The aim of this study was to identify the most stable reference gene(s) for quantification of microRNA expression analysis in uterine cervical tissues. A microarray was performed on 6 pairs of uterine cervical tissues to identify the candidate reference genes. The stability of candidate reference genes was assessed by RT-qPCR in 23 pairs of uterine cervical tissues. The identified most stable reference genes were further validated in other cohort of 108 clinical uterine cervical samples: (HR-HPV- normal, n = 21; HR-HPV+ normal, n = 19; cervical intraepithelial neoplasia [CIN], n = 47; cancer, n = 21), and the effects of normalizers on the relative quantity of target miR-424 were assessed. In the array experiment, miR-26a, miR-23a, miR-200c, let-7a, and miR-1979 were identified as candidate reference genes for subsequent validation. MiR-23a was identified as the most reliable reference gene followed by miR-191. The use of miR-23a and miR-191 to normalize expression data enabled detection of a significant deregulation of miR-424 between normal, CIN and cancer tissue. Our results suggested that miR-23a and miR-191 are the optimal reference microRNAs that can be used for normalization in profiling studies of cervical tissues; miR-23a is a novel microRNA normalizer.

Predicting preterm delivery in asymptomatic pregnant women with prior preterm delivery by measurement of cervical length, fetal fibronectin and phosphorylated insulin-like growth factor-binding protein-1

To evaluate the accuracy of cervical length measurement in combination with assessment of fetal fibronectin [FFN] and phosphorylated insulin-like growth factor-binding protein-1 [phIGFBP-1] in cervico-vaginal secretions as a predictor of preterm delivery in asymptomatic pregnant women with a history of preterm birth. 91 singleton pregnant mothers [between 22-24 weeks of gestation] with previous history of one or more unexplained preterm birth were included in the study. For all participants transvaginal sonographic measurement of cervical length was carried out together with qualitative assay of fetal fibronectin and quantitative measurement of phosphorylated insulin-like growth factor binding protein-1 in cervico-vaginal secretions. The primary outcome of the study was delivery before completed 37 weeks of gestation. There was a significant association between cervical length and the occurrence of preterm delivery [p=0.002], cervical length was 23.12 +/- 8.5mm in 33 cases who experienced preterm delivery compared to 29.34 +/- 9.4mm in 58 cases who delivered at term. Regarding fetal fibronectin assay in cervico-vaginal secretions, no statistically significant difference was found between those who delivered preterm and those who had term delivery [p=0.972]. Measurement of phosphorylated insulin-like growth factor-binding protein-1 [phIGFBP-1] in cervico-vaginal secretions showed statistically significant difference among patients who delivered preterm compared to those who did not [p=0.007]. Sensitivity, specificity, positive and negative predictive values, and positive and negative likelihood ratios were calculated for cervical length, phIGFBP-1, and for their combination, and our results demonstrated high sensitivity, specificity, positive and negative predictive values for the combined method compared with either method alone. Both cervical length and phIGFBP-1 measurement in cervico-vaginal secretions at 22-24 weeks are likely to be useful in predicting preterm delivery in asymptomatic women with a history of preterm birth and their combination increased their sensitivity, specificity, positive and negative predictive values as predictors

In vitro study of biosynthesis of protoporphyrin IX induced by Ù-Aminolevulinic acid in normal and cancerous cells of the human cervix

Bxkground. Ù-Aminolevulinic acid (ALA) is recognized as the starter in the biosynthesis of the heme group, the structural basis of cytochromes, chlorophylls, biliary pigments, and other porphyrins. It is the first intermediary in the biosynthesis of protoporphyrin IX (PpIX), and of ther heme group. PpIX is present in low concentration in normal cells, and in high concentration in tumor cells. Methods. The accumulation of protoporhyrin IX (PpIX) induced by Ù-aminolevulinic acid (ALA) was tested in two cervico-uterine cancer cell lines (HeLa and CaLo), and in normal human cervical epithelial (NHCE) cells. Results. The optimal concentration of ALA that induced maximum levels of intra- and extracellular accumulation of PpIX in both HeLa and NHCE cells was 300 µg of ALA/mL, and for CaLo cells, 150 µg/mL. The viability of HeLa, CaLo, and NHCE cells exposed to ALA measured 81, 98 and 84 percent, respectively. The optimale time for accumulation of PpIX, both intra- and extracellular, was 4 h for HeLa and NHCE cells and 5 h for CaLo cells per 24 h of expusure to optimal concentrations of ALA. After the maximum level of PpIX accumulation was reached, there was a gradual decrease until there was only a small quantity. A statistically significant difference (p <0.0001) was foundf in the accumulation of PpIX, depending on the concentrations of ALA used as well as between cervical cancer cell lines and 1:7, and for NHCE and CaLo cells. 1:5. Conclusions. These results are important for determining the usefulness of the sensitizer (PpIX)

Shedding of HIV-1 subtype E in semen and cervico-vaginal fluid.

The uneven expansion of HIV-1 subtypes in each transmitted group raises the possibility that some viruses have less/more potential by qualitative/quantitative for heterosexual transmission compared to others. In Thailand, HIV-1 subtype E is mainly spread via heterosexual route and accounts for about 95 per cent of the infected cases. To determine whether high sexual infectivity of HIV-1 subtype E is due to the presence of a virus in genital fluid, we conducted a study to characterize shedding of HIV-1 in seminal and cervico-vaginal fluids of 30 HIV-1 subtype E infected Thai couples by PCR and virus isolation methods. All subjects had no HIV-associated diseases and other sexually transmitted diseases. HIV-1 subtype E DNA was detected in 22/30 (77.33%) of cervico-vaginal and also 22/30 (77.33%) of seminal fluid samples. The isolation rate of HIV-1 from semen and cervico-vaginal secretion was 36.67 per cent and 16.67 per cent, respectively. Number of HIV-1 subtype E DNA copies in the blood is reversely correlated with the number of blood CD4+ T cells, while that in genital fluid was not related to CD4+ T cell count. An increase in shedding of HIV- DNA subtype E in female genital tract compared to other HIV subtypes reported by other investigators might be one reason to explain the rapid spread of subtype E by heterosexual transmission in Thailand.

Uso de tampones vaginales para la obtención de muestras citológicas del cuello uterino de pacientes ambulatorias

En el presente estudio, se tomó en cuenta a 139 mujeres con vida sexual activa que asistieron al consultorio externo del Servicio de Pediatría y niño sano del Hospital de Apoyo Camaná, a quienes se les solicitó su participación en el estudio, se les explicó que no era necesario someterse al examen pélvico. Seguidamente se entregó a la paciente un tampón vaginal comercial previa explicación del modo de inserción, frotis y fijación de la muestra. La aceptación del nuevo método llegó a un 76.9 por ciento siendo un resultado excelente para campañas de Screening. En el 100 por ciento de las muestras se evidenció cantidad suficiente de células cervicales para su lectura citológica válida. Este resultado es también excepcional. No se encontró ningún caso de Displasia ni células neoplásicas, se detectó 9 casos (8.1 por ciento) de inflamación leve, 10 (9 por ciento) de inflamación moderada y 14 casos (12.7 por ciento) de inflamación severa, así como un caso de candidiasis. Entre los factores asociados a la aceptación del nuevo método (edad, paridad, años de estudio aprobados, lugar de residencia), sólo la edad influyó en dicha aceptación