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Transfer of germinal vesicle to ooplasm of young mice could not rescue ageing-associated chromosome misalignment in meiosis of oocytes from aged mice.

BACKGROUND: [corrected] Transferring a germinal vesicle (GV) from an aged woman's oocyte into ooplasm from a younger woman has been proposed as a possible way to overcome the problem of age-related decline in female fertility. Here we assessed this possibility by determining whether ooplasts derived from young mice could rescue ageing-associated chromosome misalignment in meiosis of oocytes from aged mice. METHODS: Three groups of reconstructed oocytes, young GV-young cytoplast (group YY), aged GV-young cytoplast (group AY), and young GV-aged cytoplast (group YA), were created by micromanipulation and electrofusion. RESULTS: Nuclear transplantation was successful in 89.8-94.4% of GV-ooplast complexes, and maturation rate of the reconstructed oocytes was 93.5-97.9%. Confocal microscopy analysis showed a significantly higher rate (49.2%) of chromosome misalignment in ageing mice than in young mice (16.9%), and 57.1% of oocytes in group AY exhibited chromosome misalignment, while the abnormality rate in groups YY and YA was 16.3 and 16.7% respectively. Calcium imaging showed that the three groups of reconstructed oocytes exhibited a similar pattern of calcium oscillations upon stimulation with bovine sperm extracts. Fertilization rate and developmental capacity to 2-cell embryos were also similar among the three groups of oocytes. CONCLUSIONS: Our findings suggest that: (i) the ooplasm from young mice could not rescue ageing-associated chromosome misalignment in meiosis of GV from aged mice; and (ii) behaviour of chromosome alignment over metaphase spindle is predominantly determined by GV material.

Age Factors↗

IL17 signaling promotes oocyte developmental competence acquisition during maturation.

BACKGROUND: Defects in the acquisition of oocyte developmental competence during the maturation process causes subfertility or infertility in animals and humans. Understanding the regulatory mechanisms of oocyte maturation is essential for reproductive biology and medicine. Follicular fluid (FF) is an important microenvironment governing oocyte maturation. METHODS: A tandem mass tags (TMT)-based comparative FF proteomic analysis was employed to identify FF proteins that are potentially crucial for oocyte maturation. A very large number of pig and mouse oocytes (approximately 20,000) and embryos (over 13,000, including somatic cell nuclear transfer, parthenogenetic activation, and in vitro fertilization embryos) were used to investigate the effects of identified FF proteins on in vitro oocyte maturation and subsequent in vitro and in vivo embryo development. RNA sequencing, quantitative PCR, enzyme-linked immunosorbent assays, and immunofluorescence were used to study the expression patterns and action mechanisms of identified FF proteins in oocytes. In addition, intra-oocyte levels of glutathione and reactive oxygen species were measured to assess redox homeostasis. RESULTS: Interleukin 17D (IL17D) was identified as an important FF protein and it is significantly upregulated in porcine FF during oocyte maturation. IL17D promotes oocyte maturation by enhancing bidirectional communication between oocytes and cumulus cells, via upregulating CX43 expression and transzonal projections, which helps to maintain oocyte redox homeostasis and nuclear-cytoplasmic synchrony. IL17D treatment of oocytes enhances subsequent in vitro and in vivo full-term embryo development by modulating lipid metabolism and histone modification reprogramming. IL17D exerts its function via activating IL17 signaling through binding to CD93. Two other IL17 family members, IL17A and IL17F, also enhance oocyte maturation quality. IL17D displays a conserved expression pattern and function in pig and mouse oocytes. CONCLUSIONS: This study reveals the critical roles of IL17D in regulating oocyte developmental competence acquisition during maturation by activating IL17 signaling. The findings provide valuable insights into the molecular mechanisms underlining oocyte developmental potential acquisition and may help to develop methods for efficient production of oocytes for assisted reproduction.

Animals↗

The reliability of virtual organ computer-aided analysis (VOCAL) for the semiquantification of ovarian, endometrial and subendometrial perfusion.

OBJECTIVES: Three-dimensional power Doppler angiography (3D-PDA) has been largely used for the subjective assessment of vascular patterns but semiquantification of the power Doppler signal is now possible. We examined the intraobserver and interobserver reliability of the semiquantification of ovarian, endometrial and subendometrial blood flow using 3D-PDA, virtual organ computer-aided analysis (VOCAL) and shell-imaging. METHODS: 3D-PDA was used to acquire 20 ovarian and 20 endometrial volumes from 40 different patients at various stages of in vitro fertilization treatment. VOCAL was then used to delineate the 3D areas of interest and the 'histogram facility' employed to generate three indices of vascularity: the vascular index, the flow index and the vascularization flow index. Intraobserver and interobserver reliability was assessed by two-way, mixed, intraclass correlation coefficients (ICCs) and general linear modeling was used to examine for differences in the mean values between each observer. RESULTS: The intraobserver reliability for both observers was extremely high and there were no differences in reliability between the observers for measurements of both volume and vascularity within the ovary or endometrium and its shells. With the exception of the outside subendometrial shell volumes, there were no significant differences between the two observers in the mean values obtained for either endometrial or ovarian volume and vascularity measurements. The interobserver reliability of measurements was equally high throughout with all measurements obtaining a mean ICC of above 0.985. CONCLUSIONS: 3D-PDA and shell-imaging offer a reliable, practical and non-invasive method for the assessment of ovarian, endometrial and subendometrial blood flow. Future work should concentrate upon confirming the reliability of data acquisition and the validity of the technique before its predictive value can be truly tested in prospective clinical studies.

Endometrium↗

Molecular diversity of male sterility inducing and male-fertile cytoplasms in the genus Helianthus.

The organisation of mtDNA was investigated for 28 sources of cytoplasmic male sterility (CMS) and a fertile line (normal cytoplasm) of Helianthus annuus by Southern hybridisation. In addition to nine known mitochondrial genes ( atp6, atp9, cob, coxI, coxII, coxIII, 18S, 5S and nd5) three probes for the open reading frames in the rearranged area of PET1, orfH522, orfH708 and orfH873, were used. Genetic similarities of the investigat-ed cytoplasms varied between 0.3 and 1. Cluster analyses using the UPGMA method allowed the distinction of ten mitochondrial (mt) types between the 29 investigated cytoplasms. Most mitochondrial types comprise two or more CMS sources, which could not be further separated, like the PET1-like CMS sources (with the exception of ANO1 and PRR1), or ANN1/ANN2/ANN3, ANN4/ ANN5, ARG3/RIG1, BOL1/EXI1/PEF1/PEP1 and GIG1/ PET2. ANL1, ANL2 and the fertile cytoplasms are also regarded as one mitochondrial type. Unique banding patterns were only observed for ANT1 ( atp6), MAX1 ( atp6, orfH522 and orfH708) and PRR1 ( coxII). However, four of the mitochondrial types showed unique hybridisation signals: ANN4/ANN5 had characteristic bands for atp6 and orfH708, PEF1/PEP1/EXI1/BOL1 for atp6and coxII, and PET2/GIG1 for atp9. The PET1-like cytoplasms all shared the same patterns for orfH522, orfH708and cob (except ANO1). It could be demonstrated that CMS sources, like, e.g., PET2 and PEF1, are different from PET1 in mtDNA organisation and the CMS mechanism. Therefore, these CMS sources represent interesting candidates for the development of new hybrid breeding systems based on new CMS mechanisms.

Journal Article↗

Daily fecundability: first results from a new data base.

This multicenter study has produced a database of 7017 menstrual cycles contributed by 881 women. It provides improved knowledge on length and location of the "fertile window" (identified as of up to 12 days duration) and the patterns and level of daily conception probability. The day of ovulation was identified in each cycle from records of basal body temperature and mucus symptoms. By referencing days of intercourse to the surrogate ovulation markers, estimates of daily fecundability were computed either directly or by the Scwartz model, both for single and multiple acts of intercourse in the fertile window. The relationship between coital pattern and fecundability has been explored. Univariate analysis underlines the significant link with fecundability only of the woman's reproductive history.

Developed Countries↗

Menstrual pattern changes in laparoscopic sterilization patients whose last pregnancy was terminated by therapeutic abortion. A two-year follow-up study.

Women were used as their own controls in the comparison of presterilization and poststerilization menstrual patterns. Five parameters were studied: regularity of cycle length, duration and amount of flow and incidence of dysmenorrhea and intermenstrual bleeding. Three parameters in the electrocoagulation group (regularity of cycle length and duration and amount of flow) and one parameter in the tubal ring group(duration of flow) showed significant changes after sterilization. However, by controlling for the effect of previous contraceptive methods used, no significant menstrual pattern changes following sterilization were discerned in either technique group.

Abortion, Therapeutic↗

Fecal cortisol levels in free-ranging female chacma baboons: relationship to dominance, reproductive state and environmental factors.

Savannah baboons are one of the few mammalian species that do not exhibit seasonal reproduction patterns and are therefore ideally suited to study the effect of female reproductive states (cycling, pregnant, lactating) on cortisol levels independent of seasonal factors. Fecal samples from 10 free-ranging female chacma baboons (Papio hamadryas ursinus), collected during a period of 17 months, were analyzed using a steroid-extraction method. Reproductive state had a significant effect on fecal cortisol, with lowest levels found in estrous females. Fertility was not related to fecal cortisol levels; we found no significant differences between samples collected on conceptive and nonconceptive cycles. Environmental factors explained most of the variance of fecal cortisol levels. Cortisol measures were strongly correlated with seasonal differences such as daylight duration, temperature and the amount of time that baboons spent resting. We measured higher cortisol levels during winter months and suggest that this could be related to shorter resting periods and to the cold minimum ambient temperatures at this study site. Finally, we found no relationship between social rank nor the rate of agonistic interactions with basal fecal cortisol levels.

Adaptation, Physiological↗

Change in contraceptive choices and the effect of education on use of contraception at the family planning clinic of Sişli Etfal Training and Research Hospital, Istanbul, Turkey.

OBJECTIVE: To investigate change in contraceptive use and the effect of education on fertility and contraceptive attitudes at our family planning clinic. METHODS: For this retrospective review, medical charts from Sişli Etfal Training and Research Hospital Family Planning Center were obtained. Records were evaluated for 2514 women in 1997 and 2268 women in 2002. RESULTS: During the 6-year period studied, significant decreases occurred in the number of births, elective abortions, gravidity and number of living children (p < 0.001). When the patterns of method use were compared, use of intrauterine devices and of coitus interruptus diminished whereas pill and condom use increased. Parallel with the rise in educational level, intrauterine device use declined whereas pill and condom use rose significantly (p < 0.001). CONCLUSION: The sociocultural and educational levels of women affect their contraceptive choices. Policies aimed at raising the socioeconomic and educational level of women should be supported to prevent maternal and fetal complications as a result of unwanted pregnancies.

Adult↗

[Contraception and sexuality].

Contraception implies the responsible confrontation with fertility and sexuality. Contraceptive counselling deals directly or indirectly with sexuality and comprises therefore frequently sexual counselling. 25% of women consulting our Family Planning Clinic complain spontaneously about sexual difficulties. Out of these women one fifth suffer from sexual dysfunction related to the contraceptive method used. Hormonal contraception seems to include most frequently sexual dysfunction. The symptom most frequently complained about is lack of libido. There are typical patterns of correlation between sexual dysfunction, contraceptive compliance and side-effects of contraception which are of great important for contraceptive counselling.

Adolescent↗

Distinct expression pattern of cytokines in semen of men with genital infection and oligo-terato-asthenozoospermia.

PROBLEM: The objective of this study was to evaluate the possible relevance of cytokines in seminal plasma (SP) of patients with accessory gland infection and oligoterato-asthenozoospermia. METHOD OF STUDY: Semen samples were obtained by masturbation from 90 men and were examined for the presence of interleukin (IL)-2, IL-6, IL-8, IL-11 and soluble CD23 (sCD23) by enzyme-linked immunosorbent assay. Five groups were included: (1) fertile men (n = 20), (2) infertile men with varicocele and oligo-teratoasthenozoospermia (V-OTA, n = 20), (3) infertile men with genital infection and OTA (INF-OTA, n = 20), (4) infertile men with idiopathic testicular lesion and OTA (ITL-OTA, n = 20) and (5) infertile men with azoospermia (AZOO, n = 10). RESULTS: We found that the mean level of IL-2 was higher in SP from infertile men compared with SP from fertile men (P < 0.05). Mean levels of IL-6, IL-8, IL-11 in SP of INF-OTA were higher than that of all other groups (P < 0.05, P < 0.05, P < 0.001, respectively). However, no significant differences could be detected between other groups. A significant increase was noted in sCD23 levels in SP from men with ITL-OTA compared with all other groups (P < 0.01). We have not observed any correlations between IL-2, IL-6, IL-8, IL-11 and sCD23 levels in SP and semen parameters. Spearman's correlation coefficient revealed that there was a significant association between IL-6, IL-8, IL-11 levels in men with INF-OTA. CONCLUSION: The measurement of each cytokine separately in the SP of men with INF-OTA, in spite of the existing significant differences, does not have a diagnostic value in male infertility. However, a combined determination of IL-6, IL-8, IL-11 in the SP of men with genital infection and oligo-terato-asthenozoospermia may provide clinically useful information for the diagnosis of male accessory gland infection.

Adolescent↗

Hierarchical patterns of correlated mating in Acacia melanoxylon.

Pollen of acacias is transported by insects as polyads, composite pollen grains. The polyad has enough pollen grains to fertilize all ovules within a flower and hence all seed within a pod may be full sibs. Isozyme markers were used to test this hypothesis in two populations of Acacia melanoxylon R.Br. The proportions of fruit pods with multiple paternity detected in two populations were 0.08 and 0.15. The proportions of fullsib pairs within pods estimated by the sibling pair method were 1 and 0.63 for the two populations. Comparison of the diploid paternal genotypes of pods of single paternity showed that the probability of a common pollen source for a pair of pods was high within globular clusters (0.35) or within inflorescences (0.46) but declined to 0.10 or 0.25 within the tree at random. Thus the reproductive system acted to reinforce a hierarchy of paternal correlation within each tree.

Acacia↗

SPAM1 (PH-20) protein and mRNA expression in the epididymides of humans and macaques: utilizing laser microdissection/RT-PCR.

BACKGROUND: The Sperm Adhesion Molecule 1 (SPAM1) is an important sperm surface hyaluronidase with at least three functions in mammalian fertilization. Previously our laboratory reported that in the mouse, in addition to its expression in the testis, Spam1 is synthesized in the epididymis where it is found in membranous vesicles in the principal cells of the epithelium in all three regions. Since SPAM1 is widely conserved among mammals the aim of the study was to determine if its expression pattern in the epididymis is conserved in rodents and primates. METHODS: We used laser microdissection (LM)/RT-PCR on frozen and paraffin-embedded epididymal sections of humans (n = 3) and macaques (n = 2) as well as in situ transcript hybridization to determine if transcripts are present in the epididymal epithelium. Western analysis and immunohistochemistry were used to detect and confirm the protein expression, and hyaluronic acid substrate gel electrophoresis analyzed its hyaluronidase activity. An in silico analysis of the proximal promoter of SPAM1 was also performed to identify relevant putative transcription binding sites for the androgen receptor. RESULTS: We demonstrate that mRNA unique to SPAM1 is present in the principal cells of the epididymal epithelium in all individuals of both species studied. SPAM1 protein is present in all three regions of the epididymis, as well as the vas deferens, and is localized similarly to the transcripts. SPAM1 was shown to have hyaluronidase activity at pH 7.0. In the proximal promoter of SPAM1 were uncovered putative epididymal transcription factor binding sites including androgen receptor elements (AREs), consistent with epididymal expression. CONCLUSIONS: These findings allow us to conclude that epididymal SPAM1 is conserved in at least two mammalian classes, rodents and primates. This conservation of expression suggests that the protein is likely to play an important function, possibly in sperm maturation.

Adult↗

[Phase-directional management of protective plantations. I. Fundamentals].

In order to ensure the higher effectiveness, more stability and sustainability of Protective plantations, the definition of phase-directional management (PDM) of protective plantations was brought forward on the basis of management researches and practices for protective plantations. The basics of PDM is protective maturity, which is defined as the time when the protective plantations attain to the state that the protective plantations can provide effective and complete protection to the objects needed to be protected. Protective maturity has two points, initial protective maturity age (IPMA), the age of protective maturity started, and terminal protective maturity age (TPMA), the age of protective maturity ended. Three management phases of protective plantation, i.e., prematurity phase, the period from sapling or establishment to initial protective maturity, protective maturity phase, the period of protective maturity lasting, and regeneration phase, the period during regeneration and before the establishment, are divided based on the fundamental of protective maturity. Directional management of protective plantation means that all of the management techniques in each phase are directed at the aim of protective maturity, i.e., protective maturity is the direction of management of protective forests, and protective maturity is the final objective for the management of protective forests. In order to sustain the protective maturity state, corresponding measures should be conducted in each phase, according to the classification of protective plantations. In pre-maturity phase, the purpose of managing is to accelerate the protective maturity, therefore, the measures such as weed clearing, soil cultivation, irrigation, fertilization, intercropping and branch cutting etc. should be conducted in protective plantations. In maturity phase, the aim of managing is to sustain the protective maturity, i.e., the techniques (tending and thinning) of controlling the structure of protective plantations should be paid emphases. In the period of regeneration, the objective of managing is to recover the protective maturity, accordingly, the regeneration patterns and ways should be determined reasonably. Additionally, the methods of determining protective maturity, i.e., the core of the phase-directional management, are also given corresponding to farmland shelterbelt, sand-fixation forest and water and soil conservation forest. For farmland shelterbelt, IPMA can be determined according to the growth pattern of tree height under the suitable structure (porosity). For sand-fixation forest, it can be determined by the cover degree of the forest, i.e., the age when cover degree gets to pi/4 can be considered as IPMA. In the case of water and soil conservation forest, IPMA is determined by the canopy closure at the height of 1m above forest ground, which can be obtained from the hemispherical silhouettes in vertical direction. As for the TPMA, it can be described by the natural age of trees for all of above-mentioned three kinds of protective plantations, but the concrete methods for estimating the natural age of trees in each kind of forests are different.

Agriculture↗

Models relating the timing of intercourse to the probability of conception and the sex of the baby.

The probability that conception will result from intercourse on particular days of the menstrual cycle is of biological, demographic, and personal interest to many. We describe an existing model for conception as related to the timing of intercourse, and develop a method for fitting it by means of the expectation-maximization (EM) algorithm, using widely available software (GLIM). We then generalize the model to allow for effects on its parameters due to reproductively toxic exposures. A further extension allows one to address the question of whether the pattern of intercourse in a conception cycle has an influence on the sex of the resulting baby. We illustrate the methods by application to data from a prospective study of couples trying to begin a pregnancy.

Abortion, Spontaneous↗

Establishment of a capillary-cumulus model to study the selection of sperm for fertilization by the cumulus oophorus.

BACKGROUND: Spermatozoa have to traverse the cumulus oophorus before fertilization in vivo. Evidence suggests that the cumulus oophorus plays an important role in the fertilization process. We describe the establishment of a capillary-cumulus oophorus model with which to study the action of cumulus mass on the function of human spermatozoa. METHODS: Human cumulus oophorus was aspirated into a glass capillary. Spermatozoa were allowed to pass through the cumulus mass in the capillary from one end of the capillary. The spermatozoa that had traversed the mass were collected at the other end of the capillary and underwent sperm function analyses. RESULTS: Compared with those spermatozoa cultured in medium alone, spermatozoa exposed to the cumulus mass were more likely to have normal morphology, be capacitated and acrosome reacted, with a distinct motility pattern and better zona-binding capacity. CONCLUSION: A novel in vitro model for spermatozoa penetration through the cumulus oophorus was established. The model can be applied to investigate the effect of the cumulus oophorus on sperm functions.

Acrosome Reaction↗

Soil survival of Escherichia coli O157:H7 acquired by a child from garden soil recently fertilized with cattle manure.

AIMS: This investigation was conducted to determine the survival of a naturally occurring Escherichia coli O157:H7 in garden soil linked to a sporadic case of E. coli O157 infection in Minnesota. METHODS AND RESULTS: The presence and viability of E. coli O157:H7 was monitored in manure-contaminated garden soil for several weeks. Bacterial isolates were characterized using PCR and pulsed-field gel electrophoresis (PFGE). Isolates obtained from the patient and the garden plots during this investigation had indistinguishable PFGE patterns and had the same virulence factors (stx1, stx2, eaeA, ehxA). The E. coli O157:H7 levels obtained from the garden plots declined gradually for a period of 2 months, and on day 69 only one garden plot of four had detectable levels of pathogen. All plots were negative on day 92. The rate of decline in the soil samples stored at 4 degrees C was faster compared with soil samples that remained in ambient conditions, and in refrigerated storage E. coli O157:H7 could not be detected after 10 days. CONCLUSIONS: E. coli O157:H7 strains can survive on manure-amended soil for more than 2 months, and this survival could be reduced by low temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: This is one of the few reports that have investigated the survival of a proven virulent strain in naturally contaminated soil samples. This case stresses the importance of avoiding the use of raw cattle manure to amend soil for cultivation of foods, including soils in residential garden plots.

Animals↗