Influence of oocyte collection technique on initial chromatin configuration, meiotic competence, and male pronucleus formation after intracytoplasmic sperm injection (ICSI) of equine oocytes

There is a great variability in the success of horse oocyte maturation and fertilization among laboratories. This study was conducted to determine if the meiotic and developmental competence of horse oocytes could be dependent on the method of oocyte collection, i.e., aspiration of follicular fluid with a vacuum apparatus, or opening follicles and scraping the granulosa layer. Horse oocytes were recovered from abattoir ovaries by aspiration or scraping and classified as having compact (Cp), expanded (Ex), or partial (P) cumuli. In Experiment 1 (Part A in May and Part B in October), oocytes were fixed immediately after collection to assess whether the collection method influenced the initial chromatin configuration of oocytes. In Experiment 2, in vitro maturation rates of oocytes recovered by aspiration or scraping were compared. In Experiment 3, oocytes were matured in vitro and submitted to intracytoplasmic sperm injection (ICSI). Initial chromatin configuration differed according to collection method in that there was a significantly higher prevalence of diffuse chromatin within the germinal vesicle in oocytes recovered by scraping than in oocytes recovered by aspiration (29/87, 33% and 28/166, 17%, respectively; P < 0.01). Maturation of oocytes to metaphase II did not significantly differ between scraped and aspirated oocytes (56/101, 55.4 % vs. 65/106, 61.4%, respectively). The overall pronucleus formation rate after ICSI of oocytes recovered by scraping was not significantly different than that of oocytes recovered by aspiration (50/99, 52.6% vs. 50/85, 68.5 %, respectively); however, the rate of abnormal fertilization was significantly higher for oocytes collected by aspiration (14/73, 19% vs. 6/94, 6%, respectively; P <0.05). These results demonstrate that the collection method affects the population of recovered oocytes and may contribute to differences in results observed among laboratories working with horse oocytes.     

Genome-wide methylation profiling identifies hypermethylated biomarkers in high-grade cervical intraepithelial neoplasia

Epigenetic modifications, such as aberrant DNA promoter methylation, are frequently observed in cervical cancer. Identification of hypermethylated regions allowing discrimination between normal cervical epithelium and high-grade cervical intraepithelial neoplasia (CIN2/3), or worse, may improve current cervical cancer population-based screening programs. In this study, the DNA methylome of high-grade CIN lesions was studied using genome-wide DNA methylation screening to identify potential biomarkers for early diagnosis of cervical neoplasia. Methylated DNA Immunoprecipitation (MeDIP) combined with DNA microarray was used to compare DNA methylation profiles of epithelial cells derived from high-grade CIN lesions with normal cervical epithelium. Hypermethylated differentially methylated regions (DMRs) were identified. Validation of nine selected DMRs using BSP and MSP in cervical tissue revealed methylation in 63.2–94.7% high-grade CIN and in 59.3–100% cervical carcinomas. QMSP for the two most significant high-grade CIN-specific methylation markers was conducted exploring test performance in a large series of cervical scrapings. Frequency and relative level of methylation were significantly different between normal and cancer samples. Clinical validation of both markers in cervical scrapings from patients with an abnormal cervical smear confirmed that frequency and relative level of methylation were related with increasing severity of the underlying CIN lesion and that ROC analysis was discriminative. These markers represent the COL25A1 and KATNAL2 and their observed increased methylation upon progression could intimate the regulatory role in carcinogenesis. In conclusion, our newly identified hypermethylated DMRs represent specific DNA methylation patterns in high-grade CIN lesions and are candidate biomarkers for early detection.     

宫颈液基超薄细胞学检查临床应用价值探讨

目的探讨宫颈液基超薄细胞学检查（Thinp rep cytologic,TCT）和Bethesda系统（TBS）在宫颈病变诊断中的临床价值。方法对3146例本院宫颈疾病就诊者行TCT检查和TBS细胞学分类诊断,对TCT检查异常者进行阴道镜病理检查。结果异常涂片208例（占6．61％）,其中不典型鳞状细胞（ASC）68例（2．19％）,低度鳞状上皮内瘤变（LSIL）95例（3．02％）高度鳞状上皮内瘤变（HSIL）41例（1．31％）。鳞状细胞癌（SCC）4例（0．12％）。异常细胞涂片30-39岁80例（38．1％）,40—49岁55例（26．12％）,与其他年龄组比较差异有统计学意义（P〈0．05）。TCT检查与病理检查符合率分别为LSIL81％,HSIL92％,SCC100％,两者比较差异无统计学意义。结论TCT结合TBS应用于宫颈细胞涂片配合阴道镜活检,是筛查和诊断宫颈癌前病变的可靠手段。     

Polarized cultures of human airway epithelium from nasal scrapings and bronchial brushings

Airway epithelial cultures are generally derived from tracheas postmortem or from surgical specimens of nasal polyps or turbinates. Scrapings of the mucosal surface have been little used as starting material for cultures because of their low yield of epithelial cells and their contamination with mucous secretions, blood, and underlying connective tissue. For the first time, we report that human airway epithelial cells obtained from nasal scrapings or bronchial brushings can be grown in culture to produce polarized cell sheets suitable for studies of vectorial transport.     

Three-dimensional analysis of mandibular morphology in Otolemur

Euclidean distance matrix analysis (EDMA) of three-dimensional data is used here to examine mandibular morphology between two species of galagos. Otolemur crassicaudatus consumes primarily exudates, while O. garnettii is more frugivorous. Acquisition of exudates involves either gouging or scraping tree bark, and may involve different forces at the mandible than incising fruits. Previous studies of mandibular morphology in exudate-feeding primates produced mixed results, some suggesting that morphological specializations reflect adaptations for greater force at the anterior dentition, while others suggest specializations for producing a large gape. This study addresses these controversies by testing predictions associated with O. crassicaudatus generating greater force at the anterior dentition or producing a larger gape relative to O. garnettii. In addition, this study tests predictions associated with specializations of the anterior dentition in O. crassicaudatus as related to exudate-feeding. Crania and mandibles from 28 O. crassicaudatus and 17 O. garnettii were digitized in three dimensions, using 18 landmarks that summarize the shape of the jaw. Two-dimensional measurements were taken to assess incisor robusticity. All three-dimensional data were analyzed using EDMA, and bootstrap tests were executed to identify specific interlandmark differences that were driving any significant (P < 0.05) overall shape differences. Two-dimensional data were analyzed using Student's t-test for independent measures. Results revealed that there was a significant shape difference in mandibles between species, and that mandibles of O. crassicaudatus showed higher condyles, longer mandibles, decreased incisor procumbency, and greater incisor robusticity relative to O. garnettii. It is suggested that the results of the present study reflect adaptations for scraping in O. crassicaudatus rather than gouging.     

Genome-wide methylation profiling identifies hypermethylated biomarkers in high-grade cervical intraepithelial neoplasia

Epigenetic modifications, such as aberrant DNA promoter methylation, are frequently observed in cervical cancer. Identification of hypermethylated regions allowing discrimination between normal cervical epithelium and high-grade cervical intraepithelial neoplasia (CIN2/3), or worse, may improve current cervical cancer population-based screening programs. In this study, the DNA methylome of high-grade CIN lesions was studied using genome-wide DNA methylation screening to identify potential biomarkers for early diagnosis of cervical neoplasia. Methylated DNA Immunoprecipitation (MeDIP) combined with DNA microarray was used to compare DNA methylation profiles of epithelial cells derived from high-grade CIN lesions with normal cervical epithelium. Hypermethylated differentially methylated regions (DMRs) were identified. Validation of nine selected DMRs using BSP and MSP in cervical tissue revealed methylation in 63.2–94.7% high-grade CIN and in 59.3–100% cervical carcinomas. QMSP for the two most significant high-grade CIN-specific methylation markers was conducted exploring test performance in a large series of cervical scrapings. Frequency and relative level of methylation were significantly different between normal and cancer samples. Clinical validation of both markers in cervical scrapings from patients with an abnormal cervical smear confirmed that frequency and relative level of methylation were related with increasing severity of the underlying CIN lesion and that ROC analysis was discriminative. These markers represent the COL25A1 and KATNAL2 and their observed increased methylation upon progression could intimate the regulatory role in carcinogenesis. In conclusion, our newly identified hypermethylated DMRs represent specific DNA methylation patterns in high-grade CIN lesions and are candidate biomarkers for early detection.     

Functional significance of intramandibular bending in Poeciliid fishes

Substrate-feeding teleosts show multiple, independent evolutionary acquisitions of intramandibular bending (bending within the lower jaw)—a behavior that likely enhances performance when feeding on attached or encrusting food items. However, intramandibular bending has only been quantified for marine teleosts. Here, we examine substrate feeding in eight species from the order Cyprinodontiformes and quantify movements produced by the anterior jaws of four target species selected from the family Poeciliidae to represent a variety of trophic strategies. Intramandibular bending, defined here as bending between the dentary and angular–articular bones of the lower jaw, is not present in some poeciliids (i.e. Gambusia affinis), nor is it present in outgroup cyprinodontiforms (i.e. Fundulus rubrifrons). However, intramandibular bending is present in certain poeciliids (i.e. Poecilia sphenops), and can exceed 90°. Such jaw bending enables the production of a gape angle that approaches 120°, which likely allows the fish to maximize contact between the toothed tips of the jaws and the substrate during the bite. Intramandibular bending in poeciliid species is associated with specific trophic shifts: the greater the intramandibular bending in a given species, the more attached algae (periphyton) reported in its diet. This result supports the hypothesis that intramandibular bending enhances performance when feeding on encrusting food items. We predict that additional examples of functional convergence are likely to be documented in freshwater teleosts as more herbivorous species are examined, and we propose that intramandibular bending represents an excellent model system in which to examine the functional processes that underlie convergent evolution. An erratum to this article can be found at     

慢性牙周炎患者洁刮治术后牙周微生物变化及意义

目的:研究洁刮治术后慢性牙周炎可疑致病微生物检出量与临床疗效的关系。方法:随机选取我院2012年1月至2014年5月慢性牙周炎患者36例,均采用洁刮治术治疗。分别于治疗前后检测患者牙周膜附着丧失量、牙周探诊深度、出血率以及微生物数量。结果:治疗前后比较,患者牙周探诊深度与探诊出血率差异具有统计学意义(P0.05),临床附着丧失量无统计学差异(P0.05);A组治疗后中间普氏菌数量显著高于治疗前(P0.05);牙龈卟啉单胞菌、齿垢密螺旋体和福赛氏类杆菌显著低于治疗前(P0.05);B组治疗前后细菌数量无统计学差异(P0.05);牙龈卟啉单胞菌C组牙周探诊深度变化显著高于D组(P0.05);齿垢密螺旋体C组探诊出血率变化显著低于D组(P0.05);福赛氏类杆菌C组牙周探诊深度和出血率与D组存在显著性差异(P0.05)。结论:牙龈卟啉单胞菌、齿垢密螺旋体和福赛氏类杆菌数量与洁刮治术后疗效密切相关,可作为微生物指标。