Imiquimod cream and C02 laser vaporization in vulvar intraepithelial neoplasia (VIN) 2/3 treatment.

OBJECTIVES: To compare the use of topical 5% imiquimodt (IMQ) cream or CO2 laser vaponization as the treatment of vulvar inmraepithelial lesions (VIN) 2/3 and to evaluate the degrees of residual or recurrent lesions. MATERIALS AND METHODS: Twenty-nine women with VIN 2/3 were separated into two groups, according to the proposed treatments. All were submitted to collection of vulvar swabs for DNA genotyping of human papillomavirus (HPV), vulvoscopy, and biopsy of the found lesions. After treatment they were followed up in quarterly consultations to (until) possible appearance of new lesions or along one year. RESULTS: The findings were similar in effectiveness and presence of residual or recurrent lesions on the performed treatments. However, patients treated with topical 5% IMQ cream had less severe lesions in histological recurrence when compared to those submitted to the CO2 laser vaporization. CONCLUSIONS: The effectiveness of topical 5% IMQ cream was similar to that of CO2 laser vaporization. There was no difference between the treatments for the presence of residual or recurrent lesions. However, patients who received IMQ had less aggressive lesions than those submitted to the treatment with CO2 laser vaporization.

HPV genotyping and p16 expression in Xingu Indigenous Park, Brazil.

The association between high-risk human papillomavirus (HPV) genotypes and p16 expression in indigenous women from the Xingu Indigenous Park, Brazil, was unknown. This study evaluated p16 expression in women with a histological diagnosis of cervical intraepithelial neoplasia (CIN) 3 or higher and correlated this expression with HPV genotypes to determine possible discrepancies in the expression of this marker. We evaluated 37 previously collected samples with different HPV genotypes and high-grade lesions diagnosed based on cytology, histology, and colposcopy. Immunohistochemical analysis was performed using paraffin-embedded tissue sections and the CINtec® Histology Kit. p16 protein expression was investigated by immunostaining with an anti-p16 antibody. HPV genotyping was performed by reverse hybridization. The age of the study population ranged from 22-75 years (43.81 ± 15.89 years) and parity ranged from 1-11 (5.92 ± 2.58). Thirteen different HPV genotypes were found using the INNO-LiPA kit. Single and multiple infections by HPV were found with prevalence of single infections (P = 0.029). Comparison between HPV genotype and simple or multiple infections was highly significant; it was observed more HPV 52 followed by HPV 16 in single infections (P < 0.001). p16 expression was predominantly diffuse, which was observed in 91.7% of lesions, whereas 8.3% were focal (P < 0.001). HPV 52, HPV 16 and 31 were the most prevalent HPV types in high-grade CIN in these indigenous women. Diffuse p16 expression in high-grade CIN was not influenced by the viral genotype; however, more studies are necessary to further our understanding of this restricted group.

Taking the Leap: Runx1 in the Formation of Blood from Endothelium.

Blood cell formation in the embryo occurs from multiple anatomic sites and results in the production of hematopoietic stem and progenitor cells that appear in overlapping waves. The transcription factor Runx1 is involved in a dramatic step of this process, for the transition from an endothelial cell that is integrated in a monolayer to a nonadherent circulating blood cell, a process conceptually similar to the epithelial to mesenchymal cell transition. Here we will review the role of Runx1 in the so-called hemogenic endothelium. We will describe the blood cell progenitors for which Runx1 is required, the proximal upstream transcription factors and signaling events that regulate its expression, and some of its important downstream targets in the hemogenic endothelium.

Expression of human papillomavirus E6 and E7 oncoprotein mRNA in women with low-grade squamous intraepithelial lesions or less.

We verified the prevalence of human papillomavirus (HPV) E6/E7 protein mRNA expression in patients with low-grade squamous intraepithelial lesions (LSILs) and negative cervicovaginal cytology. To investigate the relationship between mRNA expression and viral infection type, we assessed genotyping in single infections. Samples from 825 women were submitted to the E6/E7 survey. We noticed a larger percentage of E6/E7 mRNA expression in the atypical squamous cells of undetermined significance (ASC-US) and LSIL cytologies. Negative results of mRNA expression were in accordance with negative cytologies. In positive cases, the infection by a single HPV type was most common, with type 16 being most prevalent. The expression of mRNA was most prevalent in ASC-US and LSIL cytologies, compared with the negative cytology. The infection by a HPV type was more frequent in cases of positive expression, with HPV type 16 being found most frequently. Patients with LSIL cytologies had a higher percentage of multiple infections.

Immunoexpression of HPV 16/18 E6 and E7 oncoproteins in high-grade cervical squamous intraepithelial lesions in HIV-positive women.

The aim of this study was to assess the immunoexpression of human papillomavirus genotypes 16 and 18 (E6 and E7) oncoproteins in cervical high-grade squamous intraepithelial lesions (HSIL) of human immunodeficiency virus (HIV)-positive women. These results were also compared to the persistence and/or recurrence of lesions after loop electrosurgical excision procedure. Cervical samples from 158 patients were divided into three groups according to the presence or absence of HSIL in women who were or were not HIV-positive. By using the tissue microarray technique, immunohistochemistry was performed to analyze the expression of HPV 16/18 E6 and E7 oncoproteins. Cervical samples from 95 HIV-positive women and 63 HIV-negative women were studied. A statistically significant difference was found in the immunoexpression of E6 and E7 oncoproteins in samples from HIV-positive women with HSIL and that of women with non-neoplastic tissue (P < 0.001). There was also a statistically significant correlation between the immunoexpression of E6 (P = 0.012) and E7 (P < 0.001) oncoproteins in lesion persistence among HIV-positive women. Within the limitations of this study, the immunoexpression of HPV 16/18 E6 and E7 oncoproteins may have prognostic value regarding lesion persistence in HIV-positive women.

Prevention, diagnosis and treatment of cervical cancer precursor lesions at the Xingu Indigenous Park, Brazil.

PURPOSE: To describe the prevention, diagnosis, and treatment of cervical cancer precursor lesions at the Xingu Indigenous Park (PIX) from 2005 to 2006. MATERIALS AND METHODS: Observational, transversal study. The research sample consisted of 503 sexually active women aged 12 years and older. The research was performed in three stages: screening, colposcopy, and surgical treatment by large loop excision of the transformation zone. RESULTS: The cytopathological screening coverage was of 99.6%. The rate of cytologic atypia was 11.7%. Together, low-grade squamous intraepithelial lesions (LSILs) and high-grade squamous intraepithelial lesions (HSILs) were observed in 4.6% of the women. The cytological examination returned a sensitivity of 54%, specificity of 97%, a positive predictive value of 88%, and a negative predictive value of 83%. In the anatomopathological examinations of biopsies, the rate of HSILs was 30.2%. The sensitivity of the anatomopathological examination of biopsies was 72.2%, the specificity was 100%, the positive predictive value was 100%, and the negative predictive value was 44.4%. CONCLUSIONS: Viable strategies for preventing, diagnosing, and treating cervical cancer precursor lesions in women from the PIX include increasing annual coverage of cytopathological examinations, early detection of cervical intraepithelial lesions, and treatment and follow-up of detected cases.

HPV infection-associated anogenital cyto-colpo-histological findings and molecular typing in HIV-positive women.

HIV and human papillomavirus (HPV) coinfection is increasing, especially in the anal canal (AC) and cervico-vaginal regions. We identified anal epithelium abnormalities related to high-risk HPV (HR-HPV) lesions in the lower genital tracts (LGTs) of HIV-positive women, described the HPV genotypes identified, and assessed the expression of E6/E7 oncogenes in coinfected patients. Ninety-eight women were enrolled in groups combining HIV status and presence or absence of HPV in the LGT. Anal and cervical smears were collected for cytology and HR-HPV assays using Cobas(®) and/or PapilloCheck(®). Samples with highly oncogenic HPV genotypes were confirmed by NucliSENS EasyQ(®). Forty-two HIV-positive (25-52; mean age 39.5) and 56 HIV-negative (18-58; mean age 35.7) patients were included. E2 and C1 groups presented AC alterations (P = 0.002); altered images for high-resolution anoscopy were higher in E1 and C2 (P < 0.001). Of the 29 women with alterations, 41.38% were HIV-negative and 58.62% were HIV-positive (P < 0.001). HIV-positive patients accounted for 29% of the anal high-grade squamous intraepithelial lesions (P = 0.015). The Cobas(®) positive result frequency was higher in three AC groups than in the other groups. There was variation in the number of HPV types in the cervico-vaginal samples among the study groups (P < 0.001). Anal cytology and anoscopy showed more altered findings in HIV-positive patients with HPV in the LGT. HR-HPV anal infections by various genotypes are common and are associated with cervical infections in HIV-positive patients. E6/E7 expression is apparently more common in the AC of HIV-positive women.

Accuracy of the cytopathology, bacterioscopy, and vaginal flora culture.

INTRODUCTION: An over-population of vaginal microorganisms causing inflammatory processes renders it difficult to properly assess the cytopathological exam that aims to screen precedent cervical lesions. On the contrary, the occurrence of the microbial flora saprophyte does not influence correct cythodiagnosis. OBJECTIVE: To assess the composition of vaginal tract aerobic microorganisms of asymptomatic women in menacme and post-menopause, and to analyze the accuracy of cytopathologic, bacterioscopic exams, and culturing of the flora. METHODS: The women were first submitted to a focused anamnestic interrogatory and then submitted to gynecological exam. A sample of the vaginal fluid was collected with a culture swab and a smear was made on two glass slides for stained bacterioscopic exam (GRAM). The collection of material was then compiled in a cytopathologic smear analysis. All women signed the free and informed consent letter and the project was approved by the Ethics Research Board of Hospital São Paulo - UNIFESP. RESULTS: Bacterioscopy and culture proved to be better than the cytopathologic exam in featuring the bacilli and cocci. The bacterioscopy provided a better detection of the presence of bacilli (p < 0.001); no statistical difference was seen between both exams with respect to the detected cocci. The beta-hemolytic Streptococcus group was of significance in post-menopausal women (p < 0.05). CONCLUSION: In this study, the bacterioscopic and culture exams of the vaginal fluid were more effective in assessing the vaginal flora and in the detection of bacilli, compared to the cytopathological exam.

Depth of glandular crypts and its involvement in squamous intraepithelial cervical neoplasia submitted to large loop excision of transformation zone (LLETZ).

BACKGROUND: The authors aimed to confirm the depth of six mm in order to achieve an optimal eradication of the lesion. MATERIALS AND METHODS: This is a retrospective observational study of 94 cervical surgical pieces from women aged 17 to 22 years with a cyto-colpo-histopathological diagnosis of high-grade squamous cervical intraepithelial neoplasia (CIN II and/or CIN III) submitted to large loop excision of transformation zone (LLETZ). The glandular crypts and margins, both exposed or not to CIN, were assessed. The compromise and the maximum depth of the glandular crypts were noticed. RESULTS: After LLETZ, 23 (24.47%) cases presented a neoplasic impairment of endocervical margin and ten (10.64%) of the ectocervical margin. The largest noticed crypt measured 4.500 mm and the shortest 0.100 mm, with an average of 2.148 mm. CONCLUSIONS: Squamous CIN more frequently show the exposure of surgical margins to LLETZ. The deeper location of glandular crypts in the cases studied was 4.500 mm, while the largest neoplastic extension was 3.000 mm.The therapeutic method depends on this knowledge.

Efficacy of vaginal use of topical estriol in postmenopausal women with urogenital atrophy.

OBJECTIVE: This study evaluates the effect of intravaginal estriol on urogenital atrophy, Pap smear parameters, colposcopy parameters and discomfort during gynecological examination. METHODS: 31 postmenopausal women who had not used hormone therapy in the previous six months were studied. All women used intravaginal estriol, 1 mg/day for 21 days. The following variables were analyzed before and after treatment: complaints of urogenital atrophy; cytological parameters, colposcopic parameters, and subjective evaluation of discomfort during gynecologic examination. RESULTS: All urogenital atrophy complaints improved after treatment. At the first visit, 45.2% of women presented a predominance of profound cells, 51.6% with predominance of intermediate cells, and 3.2% with predominance of superficial cells. At the second visit, these rates were 35.5%, 64.5%, and 0%, respectively. Evaluation of the maturation index showed that 83.9% of women had atrophic Pap smears, and 16.1% showed good estrogenic level before treatment. At the second visit, atrophic smears occurred in 12.9%, and 87.1% of women exhibited good estrogenic level (chi2 = 20.045; p = 0.000). Colposcopy showed that 71% of women had atrophic colpitis and/or petequiae before treatment, while atrophy after therapy was present in only 6.4%. The evaluation of other colposcopic parameters also improved after treatment. Great discomfort was reported by 19.4% before and by 0% after treatment. CONCLUSION: Intravaginal estriol 1 mg/day for a period of 21 days was efficient in improving urogenital atrophy, Pap smear parameters and colposcopic evaluation in postmenopausal women.

Notch signaling in mammalian hematopoietic stem cells.

Notch is a crucial cell signaling pathway in metazoan development. By means of cell-cell interactions, Notch signaling regulates cellular identity, proliferation, differentiation and apoptosis. Within the last decade, numerous studies have shown an important role for this pathway in the development and homeostasis of mammalian stem cell populations. Hematopoietic stem cells (HSCs) constitute a well-defined population that shows self-renewal and multi-lineage differentiation potential, with the clinically relevant capacity to repopulate the hematopoietic system of an adult organism. Here, we review the emergence, development and maintenance of HSCs during mammalian embryogenesis and adulthood, with respect to the role of Notch signaling in hematopoietic biology.

Immunohistochemical expression of granzyme B and vascular endothelial growth factor (VEGF) in normal uterine cervices and low and high grade squamous intraepithelial lesions.

PURPOSE: This study aimed to evaluate the immunoexpression of granzyme B and vascular endothelial growth factor (VEGF) in the variants of cervical squamous intraepithelial neoplasia. METHODS: Granzyme B immunohistochemical expression was studied in the epithelium, stroma and in both the epithelium + stroma of 142 fragments of uterine cervix; there were 34 grade 1 cervical intraepithelial neoplasias (CIN 1), 36 grade 2 cervical intraepithelial neoplasias (CIN 2), 33 grade 3 cervical intraepithelial neoplasias (CIN 3) and 39 uterine cervix fragments without abnormalities - control group. Immunoexpression of VEGF was studied in 160 uterine cervix fragments, with 43 grade 1 cervical intraepithelial neoplasias (CIN 1), 33 grade 2 cervical intraepithelial neoplasias (CIN 2), 31 grade 3 cervical intraepithelial neoplasias (CIN 3) and 53 uterine cervix fragments without abnormalities--control group. RESULTS: In the stroma, immunoexpression of granzyme B in grade 1 cervical intraepithelial neoplasias was smaller than in grade 3 cervical intraepithelial neoplasias. High VEGF immunoexpression was found in grade 3 cervical intraepithelial neoplasias while it was low in grade 1 cervical intraepithelial neoplasias and in the control group. CONCLUSION: The higher the severity of the cervical intraepithelial lesion, the higher the immunoexpression of granzyme B. A progressive increase in VEGF immunoexpression was found in the intense grade, according to the severity of the cervical intraepithelial neoplasia.

Cytopathological screening in indigenous women from Parque Indigena do Xingu.

Over the years, there have been many deaths due to cervical cancer among indigenous women of the Parque Indigena do Xingu as a consequence of low screening coverage. Since 2005, however, the coverage index of cervical lesion screening has been high: 97.6% among at-risk women in 2005 and 92.6% in 2007. Cytological alterations occurred in 12.6% and 6% of the cases in the respective years. After complete diagnosis and treatment of uterine lesions, by staff trained in lower tract pathology, negative results were seen in all cases of high-grade lesions and invasive neoplasia and no case of invasive carcinoma was detected in 2007. We conclude, therefore, that health actions have been effective in decreasing the incidence of cytological alterations and invasive carcinoma.

Uterine cervical neoplasia prevention in Parque Indigena do Xingu.

Results of preventive health measures, diagnosis and treatment applied to Parque Indigena do Xingu native women were studied. Thirty-seven cases of uterine cervical intraepithelial lesions and invasive neoplasias were treated in the local villages without referral to an advanced medical center. LEEPs were carried out in 32 women, three cold knife conizations, one vaginal hysterectomy and one Wertheim Meigs procedure. Results of 53.1% of LEEP surgical procedures did not have margin involvement by the lesions. Bleeding complications were seen in 15.6%. Regular follow-up with two or three cytologic and colposcopic tests in 32 women was carried out. All cases were negative for lesions. Five women were not followed-up due mainly to logistical reasons. Health endeavors adopted in the period 2005-2007 brought about a significant reduction of precursor lesions in this native aboriginal population without screening resources.

Fas/CD95 promoter polymorphism gene and its relationship with cervical carcinoma.

OBJECTIVE: Apoptosis is an important fail-safe control in human papillomavirus (HPV)-associated carcinogenesis. We tested the hypothesis that the A/G polymorphism at -670 of Fas promoter is associated with an increased risk for cervical cancer, using a matched case-control setting. METHODS: The material in this case-control study consisted of 91 patients with cervical carcinoma and 176 population-based control subjects, recruited between 2002 and 2004; all the ethnic Brazilian women had histologically confirmed cervical carcinoma. Control subjects were age-matched; healthy women who were selected following a negative cervical cytology and normal colposcopy. Fas genotyping was performed using a PCR-RFLP technique. RESULTS: No significant difference existed in the distribution of the Fas polymorphisms (wild, heterozygous, mutant) between the cases and controls. The heterozygous (OR: 4.85, 95% CI: 1.1-22.6) genotypes among the younger (< 48 yrs) cancer patients were almost 5-fold increased, as compared with the wild type. No such increase was observed among the patients older than 48 years. CONCLUSIONS: Our data suggest that 670A/G polymorphism in the promoter region of the death receptor Fas is associated with an increased risk of cervical cancer among Brazilian women under 48 years. The mechanisms would be the inhibition of apoptosis by Fas -670G allele-mediated down-regulation of Fas transcription.

Carbon dioxide laser treatment of Bartholin's gland cyst.

The purpose of this study is to present the results of laser surgery for Bartholin's cyst which shows to be a method that is safe, efficient, of low morbidity, and can be performed in an outpatient clinic under local anesthesia. Of the 22 cases operated on, only two had relapses which were resolved using a second procedure. The average surgery time was short, without bleeding during the operation. Complete epithelization was achieved within a maximum 4-week span with minimal or no postoperative discomfort.

Relationship between angiogenesis and grade of histologic differentiation in endometrial adenocarcinoma.

The objective of the study was to quantify vessels and to relate them to the degree of histologic differentiation in endometrial adenocarcinoma. We studied 35 cases of which ten were G1, 13 G2 and 12 G3 adenocarcinomas. The control group consisted of 11 atrophic and 10 proliferative endometria. From each case two histologic sections were obtained: one for hematoxylin-eosin staining and the other for immunohistochemical study with anti-CD34. Vessel count was performed by morphometric study. Mean vessel count was 15.3 for G1; 19 for G2 and 22.7 for G3 adenocarcinomas; in the control group it was 11.6 for atrophic and 13.2 for proliferative endometria. Slightly differentiated adenocarcinoma presented greater angiogenesis than normal and well-differentiated carcinoma. In contrast, moderately differentiated carcinoma showed greater angiogenicity as related to normal endometrium, but did not differ from other tumoral endometria.

Angiogenesis in squamous intraepithelial neoplasia of the uterine cervix in HIV-seropositive women.

OBJECTIVES: This study aimed to quantify angiogenesis in squamous intraepithelial lesions of the uterine cervix in seropositive HIV patients as well as to establish a relationship between vascular density and variations in the CD4+ lymphocyte titer and the viral load of human immunodeficiency virus (HIV). METHODS: 125 patients, 55 HIV seropositive and 70 seronegative, were allocated with respect to grade of squamous intraepithelial lesion (SIL). The obtained samples were stained with an immunohistochemical marker for CD34 antigen and vessel counts were performed in ten consecutive fields at 400x magnification. The seropositive HIV patients were distributed into groups according to the CD4+ index and HIV viral load. RESULTS: Seropositive HIV patients presented a higher mean vascular density (MVD) than the control group, even in the absence of cervical intraepithelial lesions. High- and low-grade lesions in the presence of HIV seropositivity presented higher MVD than that found in seronegative HIV patients. There was no significant variation in the MVD and CD4+ count ratio or viral RNA-HIV load, except for high-grade (H)SIL. CONCLUSIONS: Infection with HIV influenced angiogenesis of uterine cervix in the presence of squamous intraepithelial lesions and more significantly in HSIL.

Low-dose 5-fluorouracil adjuvant in laser therapy for HPV lesions in immunosuppressed patients and cases of difficult control.

The authors established a protocol for the use of 5-fluorouracil (5FU) adjuvant in lasertherapy for clinical and subclinical HPV infection in immunosuppressed patients, persistent lesions and as reinforcement treatment in cases of poor progress. Sixty-four patients were evaluated, of whom 26 were immunosuppressed, 34 presented persistent lesions and four received intravaginal reinforcement treatment with 2.5 g 5% 5FU every two weeks, or biweekly vulvar reinforcement after lasertherapy. On average, five 5FU courses were used, but in the immunossuppressed patients its use was maintained indefinitely. The rate of complete response was 66%, but the immunossuppressed patients showed less response (46.2%) when compared with the persistent lesion/reinforcement treatment group (78.9%). The responses were positive in the two groups when compared to that with no response. We deem the use of low-dose 5FU an excellent alternative in cases of difficult HPV progress, presenting a low cost and minimal side-effects.

Spatial and temporal expression pattern of Runx3 (Aml2) and Runx1 (Aml1) indicates non-redundant functions during mouse embryogenesis.

The human RUNX3/AML2 gene belongs to the 'runt domain' family of transcription factors that act as gene expression regulators in major developmental pathways. Here, we describe the expression pattern of Runx3 during mouse embryogenesis compared to the expression pattern of Runx1. E10.5 and E14.5-E16.5 embryos were analyzed using both immunohistochemistry and beta-galactosidase activity of targeted Runx3 and Runx1 loci. We found that Runx3 expression overlapped with that of Runx1 in the hematopoietic system, whereas in sensory ganglia, epidermal appendages, and developing skeletal elements, their expression was confined to different compartments. These data provide new insights into the function of Runx3 and Runx1 in organogenesis and support the possibility that cross-regulation between them plays a role in embryogenesis.