Vulvar Paget's disease and stromal invasion: Clinico-pathological features and survival outcomes.

PURPOSE: To evaluate clinico-pathological features, treatments and survival outcomes of vulvar Paget's disease (VPD). METHODS: We retrospectively reviewed VPD diagnosed between 1983 and 2018 at the Department of Surgical Sciences, Sant'Anna Hospital, Turin. Clinico-pathological characteristics and surgical treatment outcomes were investigated according to the depth of invasion. RESULTS: A total of 122 patients were identified. Eighty-seven patients were diagnosed with intraepithelial VPD, 22 with microinvasive (<=1 mm) VPD and 16 with invasive VPD. The median follow-up was 94.6 months (interquartile range 25th-75th, 26-120). Most of patients 95/122 (77%) were treated by surgery. Local recurrence was observed in 69/95 (73%) patients without significant difference between the 3 groups (p = 0.33), however, total vulvectomy showed better local control in microinvasive and invasive VPD than in intraepithelial tumors. At 120 months the cancer-specific survival was 100% for intraepithelial and microinvasive VPD versus 31% for invasive VPD (log-rank p = <0.0001) Age ≥65 years (OR: 4.17 CI 1.12-15.5, p = 0.03) and VPD's area ≥15 cm2 (OR: 5.83 CI 1.75-19.3, p = 0.004) were associated with risk of invasiveness. CONCLUSION: Microinvasive VPD has an identical prognosis to intraepithelial VPD, suggesting the omission of lymphadenectomy or adjuvant treatments are safe in this subset of patients. We recommend caution to propose medical treatment in patients who are ≥65 years old and with wide tumor area, as they are at the greatest risk of invasiveness.

Controlled ovarian stimulation and progesterone supplementation affect vaginal and endometrial microbiota in IVF cycles: a pilot study.

PURPOSE: Does controlled ovarian stimulation (COS) and progesterone (P) luteal supplementation modify the vaginal and endometrial microbiota of women undergoing in vitro fertilization? METHODS: Fifteen women underwent microbiota analysis at two time points: during a mock transfer performed in the luteal phase of the cycle preceding COS, and at the time of fresh embryo transfer (ET). A vaginal swab and the distal extremity of the ET catheter tip were analyzed using next-generation 16SrRNA gene sequencing. Heterogeneity of the bacterial microbiota was assessed according to both the Bray-Curtis similarity index and the Shannon diversity index. RESULTS: Lactobacillus was the most prevalent genus in the vaginal samples, although its relative proportion was reduced by COS plus P supplementation (71.5 ± 40.6% vs. 61.1 ± 44.2%). In the vagina, an increase in pathogenic species was observed, involving Prevotella (3.5 ± 8.9% vs. 12.0 ± 19.4%), and Escherichia coli-Shigella spp. (1.4 ± 5.6% vs. 2.0 ± 7.8%). In the endometrium, the proportion of Lactobacilli slightly decreased (27.4 ± 34.5% vs. 25.0 ± 29.9%); differently, both Prevotella and Atopobium increased (3.4 ± 9.5% vs. 4.7 ± 7.4% and 0.7 ± 1.5% vs. 5.8 ± 12.0%). In both sites, biodiversity was greater after COS (p < 0.05), particularly in the endometrial microbiota, as confirmed by Bray-Curtis analysis of the phylogenetic distance among bacteria genera. Bray-Curtis analysis confirmed significant differences also for the paired endometrium-vagina samples at each time point. CONCLUSIONS: Our findings suggest that COS and P supplementation significantly change the composition of vaginal and endometrial microbiota. The greater instability could affect both endometrial receptivity and placentation. If our findings are confirmed, they may provide a further reason to encourage the freeze-all strategy.

The combined immunodetection of AP-2alpha and YY1 transcription factors is associated with ERBB2 gene overexpression in primary breast tumors.

INTRODUCTION: Overexpression of the ERBB2 oncogene is observed in about 20% of human breast tumors and is the consequence of increased transcription rates frequently associated with gene amplification. Several studies have shown a link between activator protein 2 (AP-2) transcription factors and ERBB2 gene expression in breast cancer cell lines. Moreover, the Yin Yang 1 (YY1) transcription factor has been shown to stimulate AP-2 transcriptional activity on the ERBB2 promoter in vitro. In this report, we examined the relationships between ERBB2, AP-2alpha, and YY1 both in breast cancer tissue specimens and in a mammary cancer cell line. METHODS: ERBB2, AP-2alpha, and YY1 protein levels were analyzed by immunohistochemistry in a panel of 55 primary breast tumors. ERBB2 gene amplification status was determined by fluorescent in situ hybridization. Correlations were evaluated by a chi2 test at a p value of less than 0.05. The functional role of AP-2alpha and YY1 on ERBB2 gene expression was analyzed by small interfering RNA (siRNA) transfection in the BT-474 mammary cancer cell line followed by real-time reverse transcription-polymerase chain reaction and Western blotting. RESULTS: We observed a statistically significant correlation between ERBB2 and AP-2alpha levels in the tumors (p < 0.01). Moreover, associations were found between ERBB2 protein level and the combined high expression of AP-2alpha and YY1 (p < 0.02) as well as between the expression of AP-2alpha and YY1 (p < 0.001). Furthermore, the levels of both AP-2alpha and YY1 proteins were inversely correlated to ERBB2 gene amplification status in the tumors (p < 0.01). Transfection of siRNAs targeting AP-2alpha and AP-2gamma mRNAs in the BT-474 breast cancer cell line repressed the expression of the endogenous ERBB2 gene at both the mRNA and protein levels. Moreover, the additional transfection of an siRNA directed against the YY1 transcript further reduced the ERBB2 protein level, suggesting that AP-2 and YY1 transcription factors cooperate to stimulate the transcription of the ERBB2 gene. CONCLUSION: This study highlights the role of both AP-2alpha and YY1 transcription factors in ERBB2 oncogene overexpression in breast tumors. Our results also suggest that high ERBB2 expression may result either from gene amplification or from increased transcription factor levels.

Analysis of the discriminant ability of shorter versions of the French ADAM questionnaire.

OBJECTIVE: To investigate whether shorter versions of the ADAM test, a screening questionnaire for andropause, provide better diagnostic value than the original tool. METHODS: Five thousand and twenty-eight volunteer men aged 50-70 years attending a screening campaign for andropause, provided a fasting blood sample and completed the French ADAM test. Logistic regression analysis identified items that best predict andropause defined as serum free testosterone level below 70 ng/l. ROC curves assessed the diagnostic value of modified versions of the ADAM test, obtained by elimination of the less relevant predictors of andropause. RESULTS: Only four items of the ADAM questionnaire may account for the diagnosis of andropause. These items concerned loss of height, decrease in libido and in enjoyment of life and deterioration in work performance. Item 9 was borderline significant. The area under the ROC curve for the short versions varied slightly from 0.555 to 0.560. As expected, model 6 has a greater specificity (56.02%) than the original tool while the efficiency increased slightly (54.85%). CONCLUSION: The modified versions of the ADAM test do not provide better diagnostic value than the original tool.

No major month to month variation in free testosterone levels in aging males. Minor impact on the biological diagnosis of 'andropause'.

BACKGROUND: The measurement of bioavailable testosterone (BT) or free testosterone (FT) levels is currently considered the gold standard for the diagnosis of androgen deficiency in elderly men. While the impact of age on circulating testosterone levels (total, bioavailable and free) has been strongly documented, the existence of seasonal variations in testosterone levels remains debated. OBJECTIVE: We investigated whether seasonal variations in serum calculated free testosterone (cFT) levels may translate into variations in the prevalence of low testosterone levels. Diagnosis was on the basis of biochemical determinations and was cross-checked with the prevalence of clinical signs and symptoms of 'andropause', as assessed by the Androgen Deficiency in Aging Males (ADAM) questionnaire. METHODS: The study recruited 5028 men aged 50 years and over from September 2000 to January 2003. Their serum FT levels were assessed and they completed the French ADAM test. Men were considered eugonadal when cFT was > or =70 ng/l. The ADAM test was scored as described originally. The prevalence of 'andropause', diagnosed by the two methods, was compared throughout the year, on a month by month basis. RESULTS: The percentage of subjects with cFT levels below 70 ng/l increased significantly with age (P<0.001). Serum cFT levels (mean [SD]) varied significantly with the month of sampling (P<0.0001), the highest (88.1 [30.2] ng/l) and lowest (76.9 [28.0] ng/l) mean values occurring in April and in October, respectively. Conversely, the prevalence of testosterone deficiency (cFT<70 ng/l) reached a peak in October (45.7%) and a nadir in April (29.7%). Although the prevalence of 'andropause', based on the ADAM questionnaire, increased significantly with age (P<0.0001), no influence of the month of the year was noticed. CONCLUSIONS: Our results confirm a progressive age-related decline in FT levels. The monthly variations in serum FT values, observed throughout the year, do not show a major seasonal rhythm in elderly community-dwelling males, since the magnitude of the variations (<15%) remains marginal. This slight variation may, however, have an impact on the number of elderly men diagnosed with Partial Androgen Deficiency in Aging Males (PADAM).

Interest of the androgen deficiency in aging males (ADAM) questionnaire for the identification of hypogonadism in elderly community-dwelling male volunteers.

OBJECTIVE: To date, serum free testosterone measurement is considered to be the gold standard for the diagnosis of hypogonadism in elderly males but it is not available to all subjects suspected of a decrease in testicular function. Therefore, we evaluated whether the Androgen Deficiency in Aging Males (ADAM) questionnaire, in its original or in a modified 'quantitative' version (qADAM), could be used as a surrogate to biochemical determinations for the identification of hypogonadism in elderly males. METHODS: 5028 men, aged 50-70 years, spontaneously consulting for the assessment of their gonadal function were studied. ADAM and qADAM, allocating a value of 1 point for any positive answer to each of the 10 questions of the ADAM test, were assessed for their ability to discriminate between males with free testosterone levels below or above 70 ng/l. RESULTS: The sensitivity and specificity of the ADAM score were 81% and 21.6% respectively. The use of ADAM resulted in an appropriate classification of our population in normal or hypogonadal subjects in 44.5% of the cases. The area under the receiver operating characteristics (ROC) curve for the qADAM (0.529) revealed a highly marginal interest of this quantitative approach compared with the original scoring system. CONCLUSIONS: The ADAM test has a high sensitivity to identify aging males with low free testosterone levels. However, due to its lack of specificity, this test cannot be used as a surrogate to serum free testosterone testing for the identification of androgen deficiency in elderly, community-dwelling males.