Developmental and family history-based analysis of congenital fused labia phenotype in the captive common marmoset (Callithrix jacchus).

BACKGROUND: Congenital fused labia (CFL) is defined as a failure or significant delay in the opening of the juvenile sealed labia majora. This phenotype is known to be variably common in adult captive female marmosets but has never been investigated in detail before. MATERIALS AND METHODS: Here, we define, describe and quantify the variations in the degree of closure of the vulva in 122 captive marmosets (Callithrix jacchus) from 1.2 to 42 months old and include colony analysis. RESULTS: There was a negative correlation between the degree of labial fusion and animal age after prepubertal period (P < 0.05). CFL females had higher number CFL relatives (4.3 ± 0.6 vs 2.4 ± 0.5 for non-CFL, P < 0.05) and more external ancestors compared to non-CFL (P < 0.05). CONCLUSIONS: Our results therefore suggest that CFL phenotype is most likely associated with epigenetic effects induced by the captive environment and colony management strategy of extensive crossing of family lines to promote heterozygosity.

Text mining and natural language processing approaches for automatic categorization of lay requests to web-based expert forums.

BACKGROUND: Both healthy and sick people increasingly use electronic media to obtain medical information and advice. For example, Internet users may send requests to Web-based expert forums, or so-called "ask the doctor" services. OBJECTIVE: To automatically classify lay requests to an Internet medical expert forum using a combination of different text-mining strategies. METHODS: We first manually classified a sample of 988 requests directed to a involuntary childlessness forum on the German website "Rund ums Baby" ("Everything about Babies") into one or more of 38 categories belonging to two dimensions ("subject matter" and "expectations"). After creating start and synonym lists, we calculated the average Cramer's V statistic for the association of each word with each category. We also used principle component analysis and singular value decomposition as further text-mining strategies. With these measures we trained regression models and determined, on the basis of best regression models, for any request the probability of belonging to each of the 38 different categories, with a cutoff of 50%. Recall and precision of a test sample were calculated as a measure of quality for the automatic classification. RESULTS: According to the manual classification of 988 documents, 102 (10%) documents fell into the category "in vitro fertilization (IVF)," 81 (8%) into the category "ovulation," 79 (8%) into "cycle," and 57 (6%) into "semen analysis." These were the four most frequent categories in the subject matter dimension (consisting of 32 categories). The expectation dimension comprised six categories; we classified 533 documents (54%) as "general information" and 351 (36%) as a wish for "treatment recommendations." The generation of indicator variables based on the chi-square analysis and Cramer's V proved to be the best approach for automatic classification in about half of the categories. In combination with the two other approaches, 100% precision and 100% recall were realized in 18 (47%) out of the 38 categories in the test sample. For 35 (92%) categories, precision and recall were better than 80%. For some categories, the input variables (ie, "words") also included variables from other categories, most often with a negative sign. For example, absence of words predictive for "menstruation" was a strong indicator for the category "pregnancy test." CONCLUSIONS: Our approach suggests a way of automatically classifying and analyzing unstructured information in Internet expert forums. The technique can perform a preliminary categorization of new requests and help Internet medical experts to better handle the mass of information and to give professional feedback.

Human oocyte chromosome analysis: complicated cases and major pitfalls.

Human oocytes that remained unfertilized in programmes of assisted reproduction have been analysed cytogenetically for more than 20 years to assess the incidence of aneuploidy in female gametes. However, the results obtained so far are not indisputable as a consequence of difficulties in evaluating oocyte chromosome preparations. Because of the lack of guidelines, we decided to summarize for the first time, the possible pitfalls in human oocyte chromosome analysis. Therefore, we screened the material from our previous studies and compiled representative, complicated cases with recommendations for their cytogenetic classification. We point out that maturity and size of the oocyte are important parameters and that fixation artefacts, as well as the particular structure of oocyte chromosomes, may predispose one to misinterpretations. Moreover, phenomena related to oocyte activation and fertilization are illustrated and explained. This compilation may help to avoid major problems in future studies and contribute to a more precise, and uniform assessment of human oocyte chromosomes.

Access to genetic material: reproductive technologies and bioethical issues.

New assisted reproduction technologies provide access to, and require the handling of, genetic material. Such material includes gametes or early-stage embryos. After a selective appraisal of some of the most relevant developments facilitated by the access to germ cells and embryos, this paper identifies some major ethical challenges created by the assisted reproduction technologies, with a special focus on Germany. The rapid progress in medical technology makes it difficult to prohibit or allow assisted reproduction technologies on the basis of such traditional criteria as the protection of human life, medical indications or a preference to adhere to natural processes. Instead, a more open discussion and a flexible application of ethical principles may be more helpful, such as: (i) considering DNA as a biological data file which allows potential access to a person's biography or identity; (ii) reconciling the protection of the early embryo with patients' needs; and (iii) placing both the mother and her embryo at the centre of bioethical and legal considerations, instead of exclusively relying on ethical principles and expert opinions with regard to the embryo alone. To continue the success of assisted reproduction, more attention should be given to the 'lifeworld' philosophy, as some authors have put it.

Information needs and visitors' experience of an Internet expert forum on infertility.

BACKGROUND: Patients increasingly use health portals and Web-based expert forums (ask-the-doctor services), but little is known about the specific needs of Internet users visiting such websites, the nature of their requests, or how satisfied they are with Internet health experts. OBJECTIVE: The aim of this study was to analyze the information requests of (mostly female) patients visiting an Internet expert forum on involuntary childlessness and their satisfaction with the experts' feedback. METHODS: We posted an electronic questionnaire on a website hosting an expert forum on involuntary childlessness. The questionnaire was "activated" whenever a visitor sent a question or request to the expert forum. The survey focused on the reasons for visiting the expert forum and whether the visitors were satisfied with the experts' answers to previously posted questions. The free-text questions of visitors who answered the survey were analyzed using Atlas-ti, a software program for qualitative data analysis. RESULTS: Over a period of 6 months, 513 out of 610 visitors (84%) answered the questionnaire. The majority of respondents (65.5%) expected general information about involuntary childlessness, conception, or an evaluation of drugs. Others were concerned about their actual treatment (40.6%) and therapeutic options (28.8%). Out of 225 respondents who had previously contacted the forum, 223 has received an answer, and 123 (55.2%) were satisfied with the experts' answers. About half (105/223) of those users who had previously received an answer from the expert forum stated that they had discussed it with their own doctor. More of these users were satisfied with their subsequent care in fertility clinics than users who did not talk to their doctor about their Internet activities (93.9% vs 76.1%; P = .015). According to the qualitative analysis, many requests (n = 194) were more or less trivial, especially those for information on basic aspects of reproduction. More than one-third of visitors (n = 199) sent detailed results of diagnostic tests and asked for a first or second opinion. Requests to the expert forum were also sent in order to obtain emotional support (17%) or to complain about a doctor (15%). CONCLUSIONS: Visitors who sent their laboratory findings to receive a thorough evaluation or a second opinion had a good command of the opportunities that an expert forum offers. One important expectation of the forum was emotional support, indicating psychological needs that were not met by medical providers. Future websites must find a compromise in order to protect experts from being overwhelmed by general, nonspecific requests while supporting patients with individualized answers.

Zona pellucida characteristics and sperm-binding patterns of in vivo and in vitro produced porcine oocytes inseminated with differently prepared spermatozoa.

IVF of porcine oocytes has been carried out in many laboratories. However, polyspermic fertilization is still a major issue to be solved. It is well known that besides the nucleus, oocyte organelles and the cytoplasm have to undergo a final maturation process before they become fully competent for fertilization. Until now, it is still uncertain whether the zona pellucida (ZP) must also undergo a maturation process and what impact the maturation status may have on sperm recognition and monospermic fertilization. Our data show that the ZP undergoes biochemical changes in the final maturation phase of the oocyte prior to fertilization. During zona maturation, the induction of the acrosome reaction in spermatozoa bound to the zona pellucida shows a different time pattern. Additionally, it was shown by 2D gel electrophoresis that after maturation, ZPA moved 0.8 pI units and ZPB/ZPC 1.3 pI units in the direction of the anode, indicating increased acidity. These preliminary studies indicate that the maturation processes of the oocyte involves biochemical and functional alterations in the zona pellucida. In addition, the morphology of the porcine ZP was investigated before and after maturation at the GVI and metaphase II stage as well as 1h after onset of IVF. No significant consistent structural changes were seen between immature oocytes and those matured in vitro for 48 h. However, at 24 h, the zona structures were more similar to those in in vivo matured oocytes. This phenomenon needs to be elucidated. So far, the only way to avoid polyspermic penetration is to reduce the number of spermatozoa per oocyte used for IVF. The amount depends on the treatment of the sperm and has to be set for each individual boar.

Analysis of N-linked glycans of porcine zona pellucida glycoprotein ZPA by MALDI-TOF MS: a contribution to understanding zona pellucida structure.

The mammalian oocyte is encased by a transparent extracellular matrix, the zona pellucida (ZP), which consists of three glycoproteins, ZPA, ZPB, and ZPC. The glycan structures of the porcine ZP and the complete N-glycosylation pattern of the ZPB/ZPC oligomer has been recently described. Here we report the N-glycan pattern and N-glycosylation sites of the porcine ZP glycoprotein ZPA of an immature oocyte population as determined by a mass spectrometric approach. In-gel deglycosylation of the electrophoretically separated ZPA protein and comparison of the pattern obtained from the native, the desialylated and the endo-beta-galactosidase-treated glycoprotein allowed the assignment of the glycan structures by MALDI-TOF MS by considering the reported oligosaccharide structures. The major N-glycans are neutral biantennary complex structures containing one or two terminal galactose residues. Complex N-glycans carrying N-acetyllactosamine repeats are minor components and are mostly sialylated. A significant signal corresponding to a high-mannose type chain appeared in the three glycan maps. MS/MS analysis confirmed its identity as a pentamannosyl N-glycan. By the combination of tryptic digestion of the endo-beta-galactosidase-treated ZP glycoprotein mixture and in-gel digestion of ZPA with lectin affinity chromatography and reverse-phase HPLC, five of six N-glycosylation sites at Asn(84/93), Asn268, Asn316, Asn323, and Asn530 were identified by MS. Only one site was found to be glycosylated in the N-terminal tryptic glycopeptide with Asn(84/93.) N-glycosidase F treatment of the isolated glycopeptides and MS analysis resulted in the identification of the corresponding deglycosylated peptides.