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Cytochemical localization of glycoconjugates in the secretory epithelial cells lining the prostate gland of the miniature pig.

The aim of the study was to emphasize several specific functional aspects of the mammalian prostate gland. In this connection, the subcellular localization and characterization of glycoconjugates in the secretory epithelial cells were examined that line the prostate gland of the miniature pig, using cytochemistry, including lectin-gold methods. The results verified a diversified pattern of glycoconjugates in the structures of the secretory cells. The secretory granules and the free surface coat of the plasma membrane contained several sugar moieties such as alpha-D-Man, alpha-D-Glc, alpha-L-Fuc, beta-D-Gal, GalNAc, GlcNAc and NeuAc (sialic acid), whereas the Golgi apparatus exhibited GlcNAc and sialic acid residues. In addition, the cisterns of the endoplasmic reticulum showed alpha-D-Man and alpha-D-Glc moieties. It seems likely that the secretory glycoconjugates demonstrated contribute to favourable conditions for fertilization.

Animals↗

Evaluation of an immunodiffusion test for screening of rubella immunity.

Rubella antigen obtained by treatment with a mild neutral detergent (Brij-58) of rubella virus infected SIRC cells gave two lines of precipitate in a modified Ouchterlony test (ID), using sera collected during convalescence after rubella. One strong line was obtained with positive sera and sometimes one thin line as well. The presence of at least one line was taken as indicator of rubella immunity. The pattern in which reagents were arranged allowed all positive serum samples to give precipitates showing antibody identity with a rubella control precipitate. Among women in the fertile age screened for immunity, 75% gave precipitates by the ID test and thus were considered to be immune. This should be compared with the 85% positivity found by means of the haemagglutination-inhibition (HI) test. As the HI test occasionally gives false results due to non-removable, non-specific inhibitors, the ID test on basis of the specificity seems to be well suited to be used as a method for the selection of individuals to be vaccinated against rubella.

Adolescent↗

Termination of pregnancy with reduced doses of mifepristone. World Health Organisation Task Force on Post-ovulatory Methods of Fertility Regulation.

OBJECTIVES: To compare the abortifacient efficacy and side effects of three doses of the antiprogestin mifepristone plus prostaglandin for termination of early pregnancy. DESIGN: Randomised, double blind multicentre trial. SETTING: 11 departments of obstetrics and gynaecology and of family planning, mostly in university hospitals, in seven countries. SUBJECTS: 1182 women with an early pregnancy (menstrual delay of 7-28 days) requesting abortion. INTERVENTIONS: Single doses of 200 mg, 400 mg, or 600 mg mifepristone followed, 48 hours later, by vaginal pessary of 1 mg of the prostaglandin E1 analogue gemeprost. MAIN OUTCOME MEASURES: Outcome of treatment; duration and subjective amount of menstrual bleeding; side effects and complications; and concentrations of haemoglobin. RESULTS: Outcome was similar with the three doses of mifepristone. Of the 1151 women with known outcome, 95.5% had a complete abortion (364 (93.8%) of those given 200 mg mifepristone, 368 (94.1%) of those given 400 mg, and 367 (94.3%) of those given 600 mg), 3.7% had an incomplete abortion (14 (3.6%), 15 (3.8%), and 14 (3.6%)), 0.3% had a missed abortion (three (0.8%), one (0.3%), and none), and 0.4% had a continuing live pregnancy (two (0.5%), two (0.5%), and one (0.3%)). Of the 43 women who had incomplete abortion, 23 underwent emergency uterine curettage (usually for haemostatic purposes) and three of these women were given a blood transfusion. The numbers of reported complaints, bleeding patterns, and changes in blood pressure and haemoglobin concentrations were similar with the three treatments. CONCLUSIONS: For termination of early pregnancy a single dose of 200 mg mifepristone is as effective as the currently recommended dose of 600 mg when used in combination with a vaginal pessary of 1 mg gemeprost.

Abortifacient Agents, Nonsteroidal↗

[Borderline tumors of the ovary. New aspects on morphologic prognosis determination].

Epithelial tumours of the ovary with a tumour status between benign and malignant are classified as borderline tumours (BOT). They are particularly frequent in women between 30 and 40 years of age, emphasising the need for effecting a therapy directed at preserving fertility. To keep the risk of this therapy at a minimum it is imperative to effect accurate determination of the prognosis on the basis of an examination of the histological preparation. Conventional histology is insufficient for this purpose, thus requiring the use of additional morphological methods. In the present study the DNA of the cell nucleus was determined via quantitative cytophotometry and also via several tumour or differentiating markers by immunohistology, in healthy ovaries, 10 benign and 20 malignant ovarian tumours and in 20 borderline tumours (BOT). The DNA histograms of the tumours classified histologically as BOT, yielded distribution patterns pointing towards a benign, proliferative or malignant potency of the tumours without a correlation to conventional histology being present in each case. Immunohistological identification of the carcinoembryonal antigen (CEA) and of the blood group substances (A, B, Lewis, Lewis) produced characteristic distribution patterns which correlated with the tendency to proliferation in DNA cytophotometry. To achieve relevant determination of prognosis of BOT in individual cases, conventional histology should be complemented by additional examinations using DNA cytophotometry and immunhistology as diagnostic tools.

ABO Blood-Group System↗

Current management of complete and partial molar pregnancy.

Molar pregnancy is composed of two separate entities, partial (PHM) and complete (CHM), which are distinct in terms of epidemiology, genetics, histopathology, clinical presentation and risk of persistent gestational trophoblastic tumor (GTT). The most common presenting symptom in patients with CHM is vaginal bleeding. Approximately half the patients with CHM show signs of exuberant trophoblastic growth, with uterine enlargement and high levels of human chorionic gonadotropin (hCG). In contrast, patients with PHM usually present as though they have an incomplete or missed abortion, with bleeding, small uteri and low hCG levels. Cytogenetically, all chromosomal material in CHM is derived from the male. Hence, no fetal parts are identified. In PHM, dispermy results in a triploid conceptus, in which an abnormal fetus is present and ultimately dies. The diagnosis of CHM is usually confirmed by sonography when a vesicular pattern is noted. The ultrasound pattern in PHM is less consistent and depends on careful measurement of the gestational sac. Patients with CHM with marked trophoblastic hyperplasia, elevated hCG levels and enlarged uteri can develop significant medical complications, which should be recognized early and treated aggressively. These include acute respiratory distress syndrome, hyperthyroidism, preeclampsia and theca lutein cysts. All molar pregnancies should be evacuated promptly following a definitive diagnosis. If the patient no longer wishes to preserve her fertility, a hysterectomy will reduce the risk of developing nonmetastatic GTT. Following evacuation, careful hCG monitoring is mandatory since it is the most reliable and sensitive method for the early detection of GTT.(ABSTRACT TRUNCATED AT 250 WORDS)

Abortion, Incomplete↗

[Prognosis and fertility after conservative treatment for ovarian tumors of limited malignity: review of 68 cases].

UNLABELLED: The aim of this retrospective study was to evaluate the rate of recurrence and the reproductive outcome after surgical conservative treatment of low malignant ovarian tumors (LMOT). MATERIAL AND METHODS: Sixty-eight patients with 50 Stage I LMOT and 18 LMOT with peritoneal implants treated conservatively at institut Gustave-Roussy, between January 1969 and December 2000. Fifty-nine patients had an unilateral adnexectomy (associated twelve times with a contralateral cystectomy), Seven had a bilateral cystectomy and two an unilateral cystectomy. Five patients received adjuvant therapy. RESULTS: With a median follow-up of 71.5 months, 16 patients recurred and one had evolutive peritoneal disease. The histologic pattern of ovarian recurrences was always of borderline type. The histologic patterns of peritoneal recurrence was similar to those initially diagnosed except in one case. Peritoneal implants, exophytic tumor and serous type tumor were significatively associated with a higher 5-year recurrence rate. Recurrence was more frequent after cystectomy than unilateral adnexectomy (p = 0.13). None of patients treated conservatively recurred under the form of ovarian carcinoma. None patient died of tumor. Nineteen patients experienced 26 pregnancies: 24 were spontaneous in a median delay of 19 months. Seven infertile patients underwent ovarian stimulation. None recurred after infertility treatments. The 2-year and 5-year cumulative pregnancy rate were respectively 41.9% and 59.8%. Four patients experienced a pregnancy after a conservative treatment of their recurrence. CONCLUSION: Despite a high recurrence rate, especially in stage II and III serous LMOT, conservative treatment of LMOT does not affect survival and should be considered in young patients. Such treatment is not advised in case of invasive peritoneal implants. Spontaneous pregnancy rate is good but the frequent infertility associated with these tumors can afterwards require ovarian stimulation. Limited number of stimulation cycles is acceptable in such patients.

Adnexa Uteri↗

Epigenetic safety of in vitro maturation in PCOS: genome-wide DNA methylation profiling of cord blood from a randomized controlled trial.

BACKGROUND: In vitro maturation (IVM) provides a safer alternative to conventional in vitro fertilization (IVF) for women with polycystic ovary syndrome (PCOS) by mitigating the risk of ovarian hyperstimulation. However, concerns persist regarding whether IVM perturbs epigenetic reprogramming in the offspring. Current evidence is constrained by candidate-gene approaches or a lack of parental controls. This study aimed to evaluate the genome-wide DNA methylation safety of IVM compared with conventional IVF using a rigorous trio-based design. METHODS: This secondary epigenetic analysis was nested within a randomized controlled trial (RCT) (ClinicalTrials.gov: NCT03463772). We included 10 nuclear families (trios), comprising five IVM-conceived and five IVF-conceived singleton offspring alongside their biological parents. Both groups utilized a uniform freeze-only single-blastocyst transfer strategy to minimize hormonal confounding. Genomic DNA from umbilical cord blood (UCB) and parental peripheral blood was analyzed using reduced representation bisulfite sequencing (RRBS). Genome-wide methylation patterns and differentially methylated regions (DMRs) were subsequently compared between the groups. RESULTS: Clinical characteristics were comparable between the IVM and IVF groups. Genome-wide analyses demonstrated high concordance in UCB methylation patterns, revealing no significant differences in global CpG methylation levels or distributions across key genomic features (promoters, CpG islands, and gene bodies). Only three rare DMRs were identified in UCB (representing ~ 0.0001% of the genome), none of which mapped to imprinted or developmentally critical loci. Furthermore, methylation variability remained consistent between the groups. CONCLUSIONS: Our findings provide robust mechanistic evidence supporting the epigenetic safety of IVM. The remarkable stability of the neonatal methylome confirms that specific IVM conditions do not compromise early developmental programming, thereby endorsing IVM as a safe and viable alternative for women with PCOS. TRIAL REGISTRATION: ClinicalTrials.gov registry, NCT03463772. Registered on March 13, 2018.

Humans↗

Practice patterns of Canadian urologists in the management of stage I testicular seminoma.

OBJECTIVE: To evaluate the practice patterns of Canadian urologists in the management of stage I testicular seminoma. METHODS: A survey was conducted with a three-page questionnaire among Canadian urologists between July and November 2002. RESULTS: The overall response rate was 48%. The total number of completed and partially completed questionnaires was 198 and 212, respectively. Ninety-four responders described their practice as university-affiliated teaching centre, while 118 reported community-based or private practice. All ordered CT abdomen/pelvis with either chest x-ray or CT scan of chest for staging investigation. Only 1% would order a lymphangiogram. About one third would not offer surveillance as a management option for stage I testicular seminoma. When asked to rank, in order of preference, three management options (surveillance, adjuvant radiotherapy, and adjuvant chemotherapy) under four different clinical scenarios varying in perceived relapse risk and the presence/absence of fertility preservation concern, the majority selected adjuvant radiotherapy as the treatment of choice for a patient with high relapse risk or no fertility concern. When a patient had desire to preserve fertility as well as low relapse risk, surveillance was chosen as the preferred management strategy. There was no significant response difference between academic and community urologists. CONCLUSION: There was some variation among Canadian urologists in the management of stage I testicular seminoma. The issue of fertility preservation and perceived relapse risk were important factors influencing management decisions. There was no significant difference between academic and community-based urologists with respect to patient volume and management approaches.

Fertility↗

A general approach for identifying distant regulatory elements applied to the Gdf6 gene.

Regulatory sequences in higher genomes can map large distances from gene coding regions, and cannot yet be identified by simple inspection of primary DNA sequence information. Here we describe an efficient method of surveying large genomic regions for gene regulatory information, and subdividing complex sets of distant regulatory elements into smaller intervals for detailed study. The mouse Gdf6 gene is expressed in a number of distinct embryonic locations that are involved in the patterning of skeletal and soft tissues. To identify sequences responsible for Gdf6 regulation, we first isolated a series of overlapping bacterial artificial chromosomes (BACs) that extend varying distances upstream and downstream of the gene. A LacZ reporter cassette was integrated into the Gdf6 transcription unit of each BAC using homologous recombination in bacteria. Each modified BAC was injected into fertilized mouse eggs, and founder transgenic embryos were analyzed for LacZ expression mid-gestation. The overlapping segments defined by the BAC clones revealed five separate regulatory regions that drive LacZ expression in 11 distinct anatomical locations. To further localize sequences that control expression in developing skeletal joints, we created a series of BAC constructs with precise deletions across a putative joint-control region. This approach further narrowed the critical control region to an area containing several stretches of sequence that are highly conserved between mice and humans. A distant 2.9-kilobase fragment containing the highly conserved regions is able to direct very specific expression of a minimal promoter/LacZ reporter in proximal limb joints. These results demonstrate that even distant, complex regulatory sequences can be identified using a combination of BAC scanning, BAC deletion, and comparative sequencing approaches.

Animals↗

Fine-scale genetic structure and gene dispersal in Centaurea corymbosa (Asteraceae) I. Pattern of pollen dispersal.

Pollen dispersal was characterized within a population of the narrowly endemic perennial herb, Centaurea corymbosa, using exclusion-based and likelihood-based paternity analyses carried out on microsatellite data. Data were used to fit a model of pollen dispersal and to estimate the rates of pollen flow and mutation/genotyping error, by developing a new method. Selfing was rare (1.6%). Pollen dispersed isotropically around each flowering plant following a leptokurtic distribution, with 50% of mating pairs separated by less than 11 m, but 22% by more than 40 m. Estimates of pollen flow lacked precision (0-25%), partially because mutations and/or genotyping errors (0.03-1%) could also explain the occurrence of offspring without a compatible candidate father. However, the pollen pool that fertilized these offspring was little differentiated from the adults of the population whereas strongly differentiated from the other populations, suggesting that pollen flow rate among populations was low. Our results suggest that pollen dispersal is too extended to allow differentiation by local adaptation within a population. However, among populations, gene flow might be low enough for such processes to occur.

Centaurea↗

[Treatment of idiopathic sterility].

In the Centre for Assisted Reproduction of the Second Gynaecological and Obstetric Clinic, First Medical Faculty, Charles University and General Hospital Prague, during the period from 1993-1997, 900 couples were examined on account of sterility. In 8%, i.e. 72 couples the cause was not elucidated. The female patients were divided into groups by age, parity and period of sterility. In these groups the cumulative conception (CCR) was compared as well as the fecundity per cycle after three months without treatment, after three cycles of intrauterine insemination (IUI) and after three cycles of in vitro fertilization (IVF) or intracytoplasmic sperm injection into the oocyte (ICSI). In the group of patients with sterility shorter than three years, the cumulative conception rate after three months without treatment was 34.8%, after the subsequent three cycles of IUI 27.3% and after three cycles of IVF/ICSI 66.7%. In women with sterility extending over more than three years these results were significantly lower: without treatment CCR was 12.5%, when IUI was used it was 16.1% and after IVF/ICSI 40.5%. There were also marked differences in the age groups. Based on these results the authors elaborated an optimal individual therapeutic pattern. Treatment can be postponed by three months in women under 30 years and those where sterility persists for less than three years. If they do not become pregnant, 3-4 IUI cycles should be attempted. Even women where IUI fails have a fair chance to become pregnant by IVF ET. In women above 30 years with sterility lasting more than three years there the chance that they will become pregnant without treatment is only when secondary sterility is involved. IUI increases the probability of pregnancy in this group only slightly. If during stimulation before IUI more than four follicles grow, it is advisable to convert these cycles to IVF. In women above 39 years the authors recommend IVF or ICSI resp. as the method of first choice.

Adult↗

Grassland management for horses.

The pasture needs for horses as compared to those for farm livestock are reviewed. The differing preferences of various types of grasses and other plants, and the patterns of grazing seen on"horse-sick" pastures, are discussed. Suggestions for practical management include frequent collection and removal of droppings, the use of grazing by cattle, adequate rest from horses and the application of cattle manure. Methods for controlling weeds and renovating horse-sick paddocks are discussed and compared to ploughing up. Finally, the place of fertilisers is considered for use on pastures for horses.

Animal Feed↗

Migration and stability.

"In this endeavor to incorporate net migration in a model of population growth, we have found that when net maternity rates are held constant, birth trajectories can be traced in several examples. We have basically experimented with (a) unchanging age-specific net migration rates and also with (b) constant number of net migrants with unchanging age composition. In most cases, the birth trajectory approaches an exponential pattern of growth with two interesting exceptions. When the intrinsic rate of growth is negative, the population approaches stationarity in the second example. Also a linear trend is found in the same example, when the net reproduction rate is equal to one." (SUMMARY IN FRE AND ITA)

Age Distribution↗

Evaluation of different biopsy methods of blastomeres from 2-cell mouse embryos.

As an outgrowth of in-vitro fertilization and embryo transfer, detection of genetic and metabolic defects prior to implantation might be possible in the future. The objective for preimplantation diagnosis would be to sample a minimum of cell material of the conceptus for diagnosis prior to transfer. Different protocols for isolating individual blastomeres out of 2-cell mouse embryos were evaluated. 2-cell mouse embryos (from F1 hybrids C57B1 females x CBA males) were collected and the zona pellucida was removed by enzyme treatment (pronase), by exposure to Ca2+-Mg2+-free acid Tyrode (pH = 2.5) or by mechanical force. Individual blastomeres were obtained by exposure to an enzyme (pronase), to a chelating agent (EDTA-glycine mixture), to Ca2+-Mg2+-free PBS or after isolation by mechanical force. The biopsied blastomeres were then cultured in vitro as such or first replaced into a host zona pellucida. Evaluation was performed by culture in vitro up to the blastocyst stage and by transfer of embryos appearing morphologically normal into pseudopregnant foster mothers. A chromosomal study of the second mitotic division of the isolated blastomere was also performed. All isolation procedures had a negative impact on the in-vitro and in-vivo growth patterns of the isolated blastomeres. After culture in vitro to the blastocyst stage, different abnormalities could be observed: embryos lacking compaction, embryos with double blastocoelic cavities and embryos with no inner cell mass (trophoblastic vesicle). After replacement of the isolated blastomeres into a host zona pellucida, similar observations could be made. Chromosomal analysis did not reveal a clear influence of the different biopsy methods on the mitosis of the isolated blastomeres.

Animals↗

Integrated morphophysiological assessment of two methods for sperm selection in bovine embryo production in vitro.

Extensive work was done regarding the ability of Swim up and Percoll gradient to select functional sperm for in vitro embryo production (IVP) systems. The aim of this work was to compare Swim up and Percoll as methods of sperm selection by ultrastructural, biochemical and functional studies. Frozen-thawed semen from two bulls (Experiments 1 and 2, respectively) were treated using Swim up or Percoll discontinuous gradients. Motility, sperm membrane ultrastructure, sperm proteins, in vitro embryo production (insemination doses, cleavage, embryo yield and quality) and embryo sex ratio were scored and compared. Electron transmission microscopy of outer sperm membranes showed higher (P<0.05) percentage of sperm with lost acrosomes in Percoll treated samples compared to Swim up. A differential protein pattern was also detected. When in vitro embryo production was performed, Percoll gradient produced higher (P<0.05) number of fertilizing doses (7.6 versus 5.9, Bull 1; 13.5 versus 7.8, Bull 2) and higher sperm motility (90% versus 76.6%, Bull 1; 81.7% versus 68.3%, Bull 2) than Swim up. The percentage of cleavage (Day 3) was similar in both treatment groups, whereas embryo production rate (Day 7) was higher (39.4% versus 30.2%, Bull 1; 38% versus 32.4%, Bull 2; P<0.05) when Percoll gradient was used. The percentage of hatched embryos (Day 11) and sex ratio did not differ. Total cell counting and embryo differential staining (inner cell mass and trophoblast cells) of Day 7 embryos showed that Percoll treated sperm produced better quality embryos compared to Swim up. We concluded that Percoll had a better performance selecting sperm and an enhanced capacity for embryo production when compared with the Swim up procedure; this could be attributed to a better acrosome exocytosis, associated to the absence of certain membrane proteins.

Acrosome↗

Disease diversity and human fertility.

The existence of parasitic constraints on the evolution of life-history traits in free-living organisms has been demonstrated in several plant and animal species. However, the association between different diseases and human traits is virtually unknown. We conducted a comparative analysis on a global scale to test whether the diversity of human diseases, some of them responsible for high incidences of morbidity and mortality, were associated with host life-history characteristics. After controlling for direct confounding effects exerted by historical, spatial, economic, and population patterns and their interactions, our findings show that human fertility increases with the diversity and structure of disease types. Thus, disease control may not only lower the costs associated with morbidity, but could also contribute directly or indirectly to reductions in human population growth.

Biological Evolution↗