Identification of novel microsatellite markers flanking GJB2 gene in order to use in preimplantation genetic diagnosis of hearing loss: A comparison of whole-genome amplification and semi-nested PCR

Hearing loss is the most prevalent sensorineural disorder which can be caused by genetic factors in more than half of the cases. GJB2 mutations with the frequency of 18.7% are the most common cause of autosomal recessive non-syndromic hearing loss (ARNSHL) in the Iranian population. The aim of the current study was to genotype 100 healthy individuals for eight microsatellite markers flanking the GJB2 gene, and to study markers on ten blastomeres using semi-nested PCR and Whole-genome amplification (WGA). All microsatellite markers within 1 Mb flanking the GJB2 gene were identified. From the identified markers, four with potentially high heterozygosity values were selected. The heterozygosity indices of four newly discovered markers and four previously reported markers were calculated. The markers and the GJB2 gene were also validated on single lymphocytes and blastomeres. Totally, 77 alleles were observed in eight loci. D13S046 showed the highest polymorphism and D13S141 showed the lowest. The observed heterozygosities of all markers, except D13S141, were higher than 50%. All single cells were genotyped successfully by the two techniques. Our findings indicate a high degree of polymorphism of the selected markers. Due to the high rate of successful amplification of markers in all ten blastomeres and the low level of allelic drop out (ADO), a combination of these eight microsatellite markers in conjunction with direct mutation detection is suggested for performing preimplantation genetic diagnosis (PGD) of hearing loss due to GJB2 mutations.     

Polygenic risk scores and risk-stratified breast cancer screening: Familiarity and perspectives of health care professionals

PurposeHealth care professionals are expected to take on an active role in the implementation of risk-based cancer prevention strategies. This study aimed to explore health care professionals’ (1) self-reported familiarity with the concept of polygenic risk score (PRS), (2) perceived level of knowledge regarding risk-stratified breast cancer (BC) screening, and (3) preferences for continuing professional development.MethodsA cross-sectional survey was conducted using a bilingual—English/French—online questionnaire disseminated by health care professional associations across Canada between November 2020 and May 2021.ResultsA total of 593 professionals completed more than 2 items and 453 responded to all questions. A total of 432 (94%) participants were female, 103 (22%) were physicians, and 323 (70%) were nurses. Participants reported to be unfamiliar with (20%), very unfamiliar (32%) with, or did not know (41%) the concept of PRS. Most participants reported not having enough knowledge about risk-stratified BC screening (61%) and that they would require more training (77%). Online courses and webinar conferences were the preferred continuing professional development modalities.ConclusionThe study indicates that health care professionals are currently not familiar with the concept of PRS or a risk-stratified approach for BC screening. Online information and training seem to be an essential knowledge transfer modality.     

Heterozygous loss-of-function variants in LHX8 cause female infertility characterized by oocyte maturation arrest

PurposeThe genetic causes of oocyte maturation arrest leading to female infertility are largely unknown, and no population-based genetic analysis has been applied in cohorts of patients with infertility. We aimed to identify novel pathogenic genes causing oocyte maturation arrest by using a gene-based burden test.MethodsThrough comparison of exome sequencing data from 716 females with infertility characterized by oocyte maturation arrest and 3539 controls, we performed a gene-based burden test and identified a novel pathogenic gene LHX8. Splicing event was evaluated using a minigene assay, expression of LHX8 protein was assessed in HeLa cells, and nuclear subcellular localization was determined in both HeLa cells and mouse oocytes.ResultsA total of 5 heterozygous loss-of-function LHX8 variants were identified from 6 independent families (c.389+1G>T, c.412C>T [p.Arg1381], c.282C>A [p.Cys941]; c.257dup [p.Tyr861]; and c.180del, [p.Ser61Profs130]). All the identified variants in LHX8 produced truncated LHX8 protein and resulted in loss of LHX8 nuclear localization in both HeLa cells and mouse oocytes.ConclusionBy combining genetic evidence and functional evaluations, we identified a novel pathogenic gene LHX8 and established the causative relationship between LHX8 haploinsufficiency and female infertility characterized by oocyte maturation arrest.     

Prenatal polybrominated diphenyl ethers exposure and anogenital distance in boys from a Shanghai birth cohort

Background

Polybrominated diphenyl ethers (PBDEs) are major brominated flame retardant (BFR) chemicals with endocrine-disrupting properties. One small-scale study on humans has suggested that prenatal exposure to PBDEs is adversely related to anogenital distance (AGD) a sensitive marker for prenatal androgen exposure. The aim of the present study was to examine the associations between prenatal exposure to PBDEs and AGD among boys 0–4 years of age in a cohort study.

Long-term quality of life among localised prostate cancer survivors: QALIPRO population-based study

BackgroundTo evaluate quality of life (QoL) 10 years after treatments for localised prostate cancer (LPCa) patients in comparison with aged-matched healthy controls.MethodsLPCa patients diagnosed in 2001 were obtained from 11 French cancer registries. Controls were recruited among the general population and were matched to patients on age and geographic area. EORTC Quality of Life Questionnaire – Core 30 items, Expanded Prostate Cancer Index Composite, Hospital Anxiety and Depression Scale and Multidimensional Fatigue Inventory self-reported questionnaires were used to measure QoL, anxiety and fatigue. Patients were classified in three groups according to previous treatments: radical prostatectomy (RP), radiotherapy (RT) and radical prostatectomy and radiotherapy (RP+RT). The differences in QoL between patients and controls and according to treatment groups were evaluated.ResultsThere were 287 patients and 287 controls. There was no socio-demographic difference between patients and controls. Treatments were: RP (143), RT (78), PR+RT (33), baseline hormone therapy (49) and hormone therapy at the time of the study (34). Patients had similar levels of global QoL, anxiety, depression and fatigue as controls. They reported more urinary troubles (urinary function and incontinence) (p < 0.0001) and more sexual dysfunctions (p < 0.0001) than controls, whatever the treatment group. Worse bowel dysfunction was reported in patients treated by RT and RP+RT (p < 0.002). According to the treatments, RP groups had the worst urinary function and incontinence (p < 0.01), and reported more bowel bother when the treatment was combined with RT.ConclusionsEven though patients reported similar global QoL as control 10 years after treatment, patients reported numerous urinary and sexual dysfunctions. Patients treated with RP+RT reported cumulative sequelae of both treatments.     

Longitudinal Trends of AMS and IIEF-5 Scores in Randomly-Selected Community Men 40 to 80 Years Old: Preliminary Results

IntroductionMost of conclusions on the relationship between age and reproductive health in aging men relied on cross-sectional data.AimTo better characterize the natural degradation trajectory of reproductive health of aging men based on longitudinal data.MethodsA community cohort study was performed in randomly selected men 40 to 80 years old, initiated in 2012 and followed up in 2014 and 2016. Participants were investigated by face-to-face structured interview, including demographic information and International Index of Erectile Function (IIEF-5) and Aging Males’ Symptoms (AMS) scales.Main Outcome MeasuresThe differences among the 3 assessments of IIEF-5 and AMS were analyzed, and progression trajectories were traced.ResultsThe high degree of variability on AMS and IIEF-5 was evident across individual subjects, as was the variability within individuals. The average IIEF-5 score of 248 subjects decreased from 16.9 to 14.1 during the 4 years, and the total AMS score increased from 22.6–27.0 (P < .001). Longitudinal data, both of individuals and of groups, showed the more rapid increase or decrease on AMS or IIEF-5 scores over 4 years in the 61–70 age group than in other age groups.Clinical ImplicationThe evidence of the greatest changes on AMS and IIEF-5 scores in the 61–70 age group prompts the importance of early intervention to postpone the degradation of reproductive health.Strength & LimitationsCompared with cross-sectional data, longitudinal data can provide a more natural progression trajectory of reproductive health of aging male individuals. The low follow-up rate might affect the parameter estimation to some extent.ConclusionCohort data over 4 years’ follow-up showed more abrupt changes on AMS and IIEF-5 scores in the 61–70 age group than in other age groups.Zheng J-B, Liang Q-F, Li J-H, et al. Longitudinal Trends of AMS and IIEF-5 Scores in Randomly-Selected Community Men 40 to 80 Years Old: Preliminary Results. J Sex Med 2019;16:1567–1573.     

幼龄SD大鼠重复给药毒性试验背景数据的建立

目的:对本研究室幼龄SD大鼠4周重复给药毒性试验中正常对照组的各项指标进行统计整理,建立本研究室的数据背景资料,为儿科药物非临床安全性评价提供参考.方法:从本研究室2015-2019年进行的儿科药物重复给药毒性试验中,选取全部正常对照组幼龄SD大鼠的数据,包括体重、摄食量、生殖发育(龟头包皮腺裂开和阴道张开时间)、骨骼...     

微流控芯片技术在精子优选中的研究进展

精子优选是体外受精( in vitro fertilization, IVF) 的关键步骤,对于提高人类辅助生殖(assisted reproductive technology,ART)成功率至关重要。研究证实,目前精子常规处理方法中的离心等过程可能造成精子活性氧(reactive oxygen species,ROS)产生增加,并损伤精子DNA。而自然受精过程中,精子需要穿过一系列解剖屏障,通过“微通道”进行迁移。以小型化和自动化为特征的微流控芯片技术恰能模拟精子在女性生殖管道中通过“微通道”自然迁移这一过程,从原精中优选获得高质量的精子,这对于提高IVF效率和改善妊娠结局具有很大的临床价值。本文对微流控芯片技术在精子优选中的研究进展进行综述,并对其应用前景进行了展望。