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Cell-free DNA (Cf-DNA) fragments may constitute an easy-to-measure molecular tool for guiding the choice of care provided to infertile couples who benefit assisted reproductive technology (ART) programmes. Data on Cf-DNA levels in the seminal plasma of men with sperm alterations are scarce. The objective of the present study was to quantify the presence of Cf-DNA in semen by using a quantitative real-time PCR. We compared men with abnormal sperm characteristics (n = 21) with normospermic controls (n = 21). The PCR assay evidenced significantly higher mean Cf-DNA levels in patients with sperm abnormalities than in controls (2.09 versus 1.18 µg/ml, respectively; p = .0003). The Cf-DNA levels were notably higher in men with azoospermia (3.65 µg/ml, versus 1.34 µg/ml in matched controls; p = .03) and men with teratozoospermia (1.80 µg/ml, versus 1.29 µg/ml in matched controls; p = .008). Our data report a significant association between elevated Cf-DNA levels and sperm abnormalities. These results may open up new diagnostic and prognostic perspectives in male infertility.

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PURPOSE: To report cases of central retinal vein occlusion in otherwise healthy children showing combined genetic variants of thrombophilia. METHODS: Ophthalmological, pediatric records and genetic analyses of thrombophilia-associated variants were retrospectively reviewed in four children diagnosed with central retinal vein occlusion. Genetic screening, including Factor XII, platelet glycoprotein (GP) IIIa PlA1/A2 (rs5918), and GPIa/IIa C807T (rs1126643) and G873A (rs1062535) mutations, was performed by PCR amplification and Sanger sequencing of PCR products. The genotyping of prothrombin G20210A, Leiden Factor V G1691A, methylenetetrahydrofolate reductase (MTHFR) C677T/A1298C mutations, and plasminogen activator inhibitor-1 4G/5G polymorphisms was performed by real-time PCR with Fluorescence Resonance Energy Transfer (FRET) probes. RESULTS: The genotyping analysis identified combined genetic variants of thrombophilia in each patient. Mutations for MTHFR (C677T) and GPIIIa PlA1/A2 were detected in Case 1, mutations for MTHFR (C677T), GPIIIa PlA1/A2, and GPIa/IIa in Case 2, mutations for MTHFR (C677T) and GPIa/IIa in Case 3, and mutation for MTHFR (A12986C), GPIIIa Pl A1/A2, and GPIa/IIa in Case 4. Preventive low-dose aspirin therapy was prescribed to all patients. During a follow-up of 5 and 8 years, neither central retinal vein occlusion recurrence nor any other thrombotic event was observed in Cases 1 and 2, respectively. CONCLUSION: In otherwise healthy children presenting central retinal vein occlusion, genetic investigations for thrombophilia-associated variants should be considered, given the possible long-term benefit of aspirin prophylaxis.

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OBJECTIVE: The gynaecological environment can become contaminated by human papillomavirus (HPV) from healthcare workers' hands and gloves. This study aimed to assess the presence of HPV on frequently used equipment in gynaecological practice. METHODS: In this cross-sectional study, 179 samples were taken from fomites (glove box, lamp of a gynaecological chair, gel tubes for ultrasound, colposcope and speculum) in two university hospitals and in four gynaecological private practices. Samples were collected with phosphate-buffered saline-humidified polyester swabs according to a standardised pattern, and conducted twice per day for 2 days. The samples were analysed by a semiquantitative real-time PCR. Statistical analysis was performed using Pearson's χ(2) test and multivariate regression analysis. RESULTS: Thirty-two (18%) HPV-positive samples were found. When centres were compared, there was a higher risk of HPV contamination in gynaecological private practices compared with hospitals (OR 2.69, 95% CI 1.06 to 6.86). Overall, there was no difference in the risk of contamination with respect to the time of day (OR 1.79, 95% CI 0.68 to 4.69). When objects were compared, the colposcope had the highest risk of contamination (OR 3.02, 95% CI 0.86 to 10.57). CONCLUSIONS: Gynaecological equipment and surfaces are contaminated by HPV despite routine cleaning. While there is no evidence that contaminated surfaces carry infectious viruses, our results demonstrate the need for strategies to prevent HPV contamination. These strategies, based on health providers' education, should lead to well-established cleaning protocols, adapted to gynaecological rooms, aimed at eliminating HPV material.

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BACKGROUND: Human papillomavirus (HPV) self-sampling (self-HPV) is valuable in cervical cancer screening. HPV testing is usually performed on physician-collected cervical smears stored in liquid-based medium. Dry filters and swabs are an alternative. We evaluated the adequacy of self-HPV using two dry storage and transport devices, the FTA cartridge and swab. METHODS: A total of 130 women performed two consecutive self-HPV samples. Randomization determined which of the two tests was performed first: self-HPV using dry swabs (s-DRY) or vaginal specimen collection using a cytobrush applied to an FTA cartridge (s-FTA). After self-HPV, a physician collected a cervical sample using liquid-based medium (Dr-WET). HPV types were identified by real-time PCR. Agreement between collection methods was measured using the kappa statistic. RESULTS: HPV prevalence for high-risk types was 62.3% (95%CI: 53.7-70.2) detected by s-DRY, 56.2% (95%CI: 47.6-64.4) by Dr-WET, and 54.6% (95%CI: 46.1-62.9) by s-FTA. There was overall agreement of 70.8% between s-FTA and s-DRY samples (kappa = 0.34), and of 82.3% between self-HPV and Dr-WET samples (kappa = 0.56). Detection sensitivities for low-grade squamous intraepithelial lesion or worse (LSIL+) were: 64.0% (95%CI: 44.5-79.8) for s-FTA, 84.6% (95%CI: 66.5-93.9) for s-DRY, and 76.9% (95%CI: 58.0-89.0) for Dr-WET. The preferred self-collection method among patients was s-DRY (40.8% vs. 15.4%). Regarding costs, FTA card was five times more expensive than the swab (~5 US dollars (USD)/per card vs. ~1 USD/per swab). CONCLUSION: Self-HPV using dry swabs is sensitive for detecting LSIL+ and less expensive than s-FTA. TRIAL REGISTRATION: International Standard Randomized Controlled Trial Number (ISRCTN): 43310942.

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Our objective was (i) to assess if a self-collected test for human papillomavirus (HPV) may serve as a primary cervical cancer screening method in a low-resource setting, (ii) to evaluate its implication in a screen and treat approach and (iii) to identify the most eligible age group in a screening program. Women were recruited through a cervical cancer screening campaign conducted in Cameroon. Written and oral instructions were given to participants by a health-care professional to carry out an unsupervised self-collected HPV-test (Self-HPV), followed by a physician-collected cervical sample for HPV testing (Physician-HPV) and cytology. Differences in performance between Self-HPV versus Physician-HPV and their ability to detect abnormal cytology results (ASC-US+) were evaluated. Descriptive analyses were used to examine the correlation between HPV positivity and cervical abnormalities by age. A sample of 789 women was prospectively enrolled. HPV prevalence was 14.6% and 12.7% for Self-HPV and Physician-HPV, respectively (Cohen's kappa = 0.74). HPV positivity by cytological diagnosis for ASC-US+ was similar with the two tests. positive predictive value of the Self-HPV for ASC-US+ was 20.4; odds ratio and number needed to treat were 6.5 (3.2-13.4) and 6 (4.2-10.9), respectively. We observed a trend of increasing cytological abnormalities in 30-49 year-old women and a concomitant trend of decreasing HPV prevalence supporting that this age group might be the most eligible group for screening. In conclusion, Self-HPV can be used as a primary screening test but needs to be followed by a triaging test that would identify the subset of women affected by clinically significant precancer or cancer.

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BACKGROUND: To evaluate if human papillomavirus (HPV) self-sampling (Self-HPV) using a dry vaginal swab is a valid alternative for HPV testing. METHODS: Women attending colposcopy clinic were recruited to collect two consecutive Self-HPV samples: a Self-HPV using a dry swab (S-DRY) and a Self-HPV using a standard wet transport medium (S-WET). These samples were analyzed for HPV using real time PCR (Roche Cobas). Participants were randomized to determine the order of the tests. Questionnaires assessing preferences and acceptability for both tests were conducted. Subsequently, women were invited for colposcopic examination; a physician collected a cervical sample (physician-sampling) with a broom-type device and placed it into a liquid-based cytology medium. Specimens were then processed for the production of cytology slides and a Hybrid Capture HPV DNA test (Qiagen) was performed from the residual liquid. Biopsies were performed if indicated. Unweighted kappa statistics (к) and McNemar tests were used to measure the agreement among the sampling methods. RESULTS: A total of 120 women were randomized. Overall HPV prevalence was 68.7% (95% Confidence Interval (CI) 59.3-77.2) by S-WET, 54.4% (95% CI 44.8-63.9) by S-DRY and 53.8% (95% CI 43.8-63.7) by HC. Among paired samples (S-WET and S-DRY), the overall agreement was good (85.7%; 95% CI 77.8-91.6) and the κ was substantial (0.70; 95% CI 0.57-0.70). The proportion of positive type-specific HPV agreement was also good (77.3%; 95% CI 68.2-84.9). No differences in sensitivity for cervical intraepithelial neoplasia grade one (CIN1) or worse between the two Self-HPV tests were observed. Women reported the two Self-HPV tests as highly acceptable. CONCLUSION: Self-HPV using dry swab transfer does not appear to compromise specimen integrity. Further study in a large screening population is needed. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01316120.

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Cryostat sections of E8.5 to E9.5 mouse embryos were hybridized with 35S-labelled RNA probes to urokinase-type and tissue-type plasminogen activator (uPA and tPA, respectively) mRNA. The sections were screened for expression of either gene. Two main features emerge from the results. (a) The stage of initial detection is different for each transcript. The uPA mRNA is first detected in cephalic mesenchyme at E8.5 when tPA mRNA is already widely distributed in tissues derived from ectoderm and mesoderm; later, the uPA mRNA transcripts were found throughout most mesodermic tissues, (b) Each gene presents a different pattern of expression. The uPA is restricted to single cells or to small groups of cells within tissues; this distribution suggests its involvement in cell migratory mechanisms. On the other hand, tPA was detected in most tissues, with variable intensities. Its expression gains complexity while organogenesis proceeds. This pattern supports the hypothesis that regulatory mechanisms other than a direct gene regulation are involved. In extraembryonic tissues, uPA and tPA genes are constantly expressed at a high level in trophoblastic giant cells and parietal endoderm, respectively. Our results confirm the presence of plasminogen activator during embryonic development and provide detailed picture of the plasminogen activator gene expression in mouse organogenesis.