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At least 379 records · Page 21Linked to original sources
PubMed2003

Serial analysis of gene expression in turkey sperm storage tubules in the presence and absence of resident sperm.

Turkey sperm lose viability within 8-18 h when stored as liquid semen using current methods and extenders. In contrast, turkey hens maintain viable, fertile sperm in their sperm storage tubules (SST) for 45 or more days following a single insemination. Our long-term objectives are to identify and characterize differentially expressed genes that may underlie this prolonged sperm storage and then use this information to develop improved methods for storing liquid turkey semen. We employed serial analysis of gene expression (SAGE) to compare gene expression patterns in turkey SST recovered from hens after artificial insemination (AI) with extended semen (sperm AI) or extender alone (control AI). We constructed two separate SAGE libraries with SST RNA obtained from sperm and control AI hens. We used these libraries to generate 95,325 ten-base pair SAGE tags. These 95,325 tags represented 27,430 unique genes. The sperm and control AI libraries contained 47,663 and 47,662 tags representing 18,030 and 19,101 putative unique transcripts, respectively. Approximately 1% of these putative unique genes were differentially expressed (P<0.05) between treatments. Tentative annotations were ascribed to the SAGE tag nucleotide sequences by comparing them against publicly available SAGE tag and cDNA sequence databases. Based on its SAGE tag nucleotide sequence, we cloned a partial turkey avidin cDNA and confirmed its up-regulation in the sperm AI SST. The bioinformatics and experimental procedures employed to clone turkey avidin and confirm its differential expression represent a useful paradigm for analyzing SAGE tag data from relatively uncharacterized model systems.

Animals↗
PubMed1999

Analysis of time-to-pregnancy data.

Statistical models for time-to-pregnancy data are usually formulated in discrete time. Two approaches were surveyed, including methods of accounting for both known heterogeneity (covariates, possibly time-dependent) and unknown heterogeneity (frailty). Delicate censoring and truncation patterns arose for prospective and, particularly, retrospective sampling designs. The inclusion of several pregnancies per couple presents new challenges and possibilities.

Cross-Sectional Studies↗
PubMed1990

Amplification of the ZFY and ZFX genes for sex identification in humans, cattle, sheep and goats.

We describe a quick and efficient method of determining the sex of DNA samples from humans, cattle, sheep, and goats. Using universal primers we have amplified 447 or 445 bp fragments from male or female genomic DNA corresponding to the ZFY or ZFX genes. Restriction fragment length polymorphism (RFLP) analysis of the fragments yielded specific banding patterns between the two sexes in these species. Horse, pig and rainbow trout fragments did not show RFLPs with the group of enzymes chosen for the other species. Use of this method would allow animal breeders to determine the sex of preimplantation embryos before transfer to recipient females. Human embryos at risk for sex-linked disorders could be sexed soon after in vitro fertilization (IVF) and options considered before implantation.

Animals↗
PubMed1998

Annexin V labelling and terminal transferase-mediated DNA end labelling (TUNEL) assay in human arrested embryos.

Confocal laser scanning microscopy was used to observe human arrested and fragmented preimplantation embryos obtained by in-vitro fertilization. Observation of the cellular actin cortex and chromatin showed a high frequency of embryos with blastomeres exhibiting two or more nuclei, while others had nuclei displaying chromatin condensation and fragmentation patterns. Many of the abnormal chromatin images could be due to the process of programmed cell death (apoptosis). The possible link between abnormalities of the blastomeres and apoptosis was investigated using two detection methods for cells undergoing apoptosis. Detection of phosphatidylserine exposure was performed using annexin V; the chromosomal breakdown preceding the nuclear collapse of apoptotic nuclei was tested using the terminal transferase-mediated DNA end labelling (TUNEL) assay. Annexin V staining was observed in all arrested and/or fragmented human embryos, but not in cryopreserved embryos which continued to develop normally after thawing. The TUNEL assay was positive in 30% (15/50) of arrested embryos, all of which had cytoplasmic fragments. In contrast, embryos showing regular size blastomeres without fragments were TUNEL negative.

Actins↗
PubMed1995

[Experimental induction of abnormalities by the instillation of ethanol in embryonated eggs of chickens (Gallus gallus)].

The teratogenic effect of alcohol on chick embryos has been confirmed by many investigators. However, how this occurs is unknown. The aim of this study was to establish a teratogenic pattern of alcohol effects, on the first stages of development in avians. Fertilized eggs were infused through the air space of the shell on day 0, with ethanol in concentrations of 20%, 40% and 60%. The control group was infused with 0.1 ml of NaCl at 0.9%. At a second stage, the eggs were treated on the 4th day of incubation, using the same method. In both groups the eggs were removed on the 11th day of incubation. The teratological manifestations that appeared more frequently were evisceration, haemorrhagic embryos, oedema, cranial deformities, lack of eyes, and umbilical hernia, showing every embryo a clear reduction in size and body weight.

Abnormalities, Drug-Induced↗
PubMed2003

Neurons expressing the highest levels of gamma-synuclein are unaffected by targeted inactivation of the gene.

Homologous recombination in ES cells was employed to generate mice with targeted deletion of the first three exons of the gamma-synuclein gene. Complete inactivation of gene expression in null mutant mice was confirmed on the mRNA and protein levels. Null mutant mice are viable, are fertile, and do not display evident phenotypical abnormalities. The effects of gamma-synuclein deficiency on motor and peripheral sensory neurons were studied by various methods in vivo and in vitro. These two types of neurons were selected because they both express high levels of gamma-synuclein from the early stages of mouse embryonic development but later in the development they display different patterns of intracellular compartmentalization of the protein. We found no difference in the number of neurons between wild-type and null mutant animals in several brain stem motor nuclei, in lumbar dorsal root ganglia, and in the trigeminal ganglion. The survival of gamma-synuclein-deficient trigeminal neurons in various culture conditions was not different from that of wild-type neurons. There was no difference in the numbers of myelinated and nonmyelinated fibers in the saphenous nerves of these animals, and sensory reflex thresholds were also intact in gamma-synuclein null mutant mice. Nerve injury led to similar changes in sensory function in wild-type and mutant mice. Taken together, our data suggest that like alpha-synuclein, gamma-synuclein is dispensable for the development and function of the nervous system.

Animals↗
PubMed1994

Enzymes of purine salvage and catabolism in the mouse preimplantation embryo measured by high performance liquid chromatography.

The activities of five enzymes involved in purine salvage and catabolism--hypoxanthine phosphoribosyl transferase (HPRT), adenine phosphoribosyl transferase (APRT), adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and guanase--were measured in mouse embryo extracts, from the one-cell to the blastocyst stage. Xanthine oxidase activity was not detected. The analyses were performed using high performance liquid chromatography and the enzymes showed different patterns of activity during development. Activities of HPRT, APRT and PNP were low before morula formation, and then increased until the blastocyst stage. ADA and guanase showed high activities after fertilization; guanase activity decreased sharply after the two-cell stage and ADA activity decreased sharply after the morula stage. Blastocyst formation was accompanied by a further decline in activity of both enzymes. The methods used may be suitable for measuring these enzymes in single human embryos, or in biopsies derived from them.

Adenine Phosphoribosyltransferase↗
PubMed1991

Fertilizing capacity of epididymal and testicular spermatozoa microinjected under the zona pellucida of the mouse oocyte.

Mouse spermatozoa were obtained from the testis and caput, corpus, and cauda epididymis incubated for 2 h in capacitation medium and a single spermatozoon from the capacitated samples microinjected into the pervitelline space of mature mouse oocytes. Spermatozoa from the testis were unable to fertilize oocytes and few spermatozoa from the caput were capable of fertilization (0-7% depending on the method of preparation). Similar fertilization rates (30-45%) were obtained with spermatozoa with forward progressive motility from the proximal and distal corpus and the cauda epididymis, but those that were vibratory or with local circular motility had significantly reduced fertilizing capacity (6-10%). The capacity of spermatozoa from the different regions of the testis and epididymis to bind to zona-free oocytes followed the same pattern as fertilization rate after microinjection. There was a progressive increase in acrosome reactions after 2 h incubation in capacitation medium in samples obtained from the testis to the cauda epididymis. Maturation of the capacity to bind to the perivitelline membrane and to develop forward progressive motility, rather than the capacity to acrosome react, appears to govern the fertilization of oocytes in which the zona has been bypassed by microinjection. These characteristics are obtained in the proximal segment of the corpus. However, there was evidence that the embryo developmental capacity of oocytes fertilized by spermatozoa from the higher segments of the epididymis is reduced, particularly in oocytes fertilized by caput spermatozoa. These observations suggest that sperm microinjection may have only a limited benefit for improving fertilization rates for men with high epididymal obstructive azoospermia.

Acrosome↗
PubMed2004

On the frequency of intercourse around ovulation: evidence for biological influences.

BACKGROUND: Intercourse in mammals is often coordinated with ovulation, for example through fluctuations in libido or by the acceleration of ovulation with intercourse. Such coordination has not been established in humans. We explored this possibility by examining patterns of sexual intercourse in relation to ovulation. METHODS: Sixty-eight sexually active North Carolina women with either an intrauterine device or tubal ligation provided data for up to three menstrual cycles. These women collected daily urine specimens and kept daily diaries of intercourse and menstrual bleeding. Major estrogen and progesterone metabolites excreted in urine were used to identify the day of ovulation. The fertile days of the cycle were defined as the 6 consecutive days ending with ovulation. Women contributed a total of 171 ovulatory cycles. Menstrual bleeding days were excluded from analysis. RESULTS: The frequency of intercourse rose during the follicular phase, peaking at ovulation and declining abruptly thereafter. The 6 consecutive days with most frequent intercourse corresponded with the 6 fertile days of the menstrual cycle. Intercourse was 24% more frequent during the 6 fertile days than during the remaining non-bleeding days (P < 0.001). CONCLUSIONS: There apparently are biological factors that promote intercourse during a woman's 6 fertile days.

Adult↗
PubMed1995

Consistency and variation in fertility investigations in Europe.

A questionnaire survey among the teaching departments of obstetrics and gynaecology in Western Europe (response rate 71%) revealed only weak adherence to the World Health Organization recommendations for the standard investigation of the infertile couple. Both general and specific examinations were applied more frequently in the female than in the male partner. Although semen analysis and the ascertainment of ovulation were standard in virtually all departments, the criteria for normal semen and the methods used for the detection of ovulation varied greatly among both departments and countries. There were also large differences among countries in the preferred standard method for testing tubal patency. The data suggest that fertility investigations are based more on tradition and personal preferences than on the demonstrated utility of its components.

Europe↗
PubMed1997

Genetics of reproduction and its evolutionary significance.

PROBLEM: To provide an evolutionary perspective on reproductive problems. METHOD OF STUDY: A broad range of evolutionary and reproductive data in the literature were analyzed. RESULTS: Evolutionary considerations provide some insight into two aspects of human reproduction. First, the inefficiency of the reproductive process (20-35% of fertilized ova yield successful pregnancies) may reflect the stress placed on the reproductive process by the relatively short evolutionary time between the hunter--gatherer and settled agricultural patterns of human life. Second, the immaturity of the immune response of the human newborn is most likely a consequence of its delivery while still in the embryonic/fetal stage of development. Delivery at this time is necessitated by the rapid development of the human brain in utero. CONCLUSIONS: The attempts to develop reproductive technologies should take into account the fundamental limits on the human reproductive process. A variety of in utero preventive and therapeutic techniques should be developed to provide the newborn with enhanced resistance to infectious diseases or to correct congenital defects. All new procedures should be thoroughly investigated and proved before being put into clinical practice.

Animals↗
PubMed1997

Norplant implants in Ethiopia.

This is a cross-sectional study involving 364 clients who have had Norplant implants insertion for contraception at the Gandhi Memorial Hospital (GMH) during the second half of 1994. It was part of the pilot project designed to introduce Norplant in Ethiopia after evaluating the safety, efficacy, tolerance, acceptability and reversibility of the method. The vast majority of the clients were found to be married, housewives, multiparae, relatively educated, urban dwellers, Christian by faith and in their reproductive age group with a mean age of 27.9 +/- 5.7 years. A significant number of them used one or more types of contraceptive methods prior to their switch to Norplant (P < 0.05). A third of the patients were lactating at the time of the Norplant insertion. The time taken to insert the implants was 12.5 +/- 4.3 minutes. The mean duration since the insertion is computed to be 25.3 +/- 4.2 months. Their main source of information was from the medical personnel. The acceptability and continuation rate of the method was 93.9% and 95.1%, respectively. The fertility-reversibility among those who desired to have childbirth was achieved in 72.3% and 84.1% at one and two years, respectively. Adverse effects of Norplant were registered in a quarter of the study population and included abnormal menstruation, headache, weight changes and pain at the site of implantation. These were not influenced by age, body weight, parity, duration of usage of the method, blood pressure patterns and previous use of contraception (P > 0.05). We have not encountered any method failure in the last 30 months. The study concludes that Norplant is safe, effective, acceptable, and reversible long-term birth control method which can be used in our setting in parallel with other available options.

Adolescent↗
PubMed1984

Aboriginal marriage and survival.

Patterns of marriage among the aboriginal population of Australia are investigated. The author attempts to determine "the economic significance of a method of family formation in which child-bearing is an essential component of the process....[and discusses the impact of] change in the micro-economies of Aboriginal households and families." Probabilities of change in marital status are also considered. Data are from the 1981 census.

Australia↗
PubMed2005

Low levels of linkage disequilibrium in wild barley (Hordeum vulgare ssp. spontaneum) despite high rates of self-fertilization.

High levels of inbreeding cause populations to become composed of homozygous, inbred lines. High levels of homozygosity limit the effectiveness of recombination, and therefore, retard the rate of decay of linkage (gametic phase) disequilibrium (LD) among mutations. Inbreeding and recombination interact to shape the expected pattern of LD. The actual extent of nucleotide sequence level LD within inbreeding species has only been studied in Arabidopsis, a weedy species whose global range has recently expanded. In the present study, we examine the levels of LD within and between 18 nuclear genes in 25 accessions from across the geographic range of wild barley, a species with a selfing rate of approximately 98%. In addition to examination of intralocus LD, we employ a resampling method to determine whether interlocus LD exceeds expectations. We demonstrate that, for the majority of wild barley loci, intralocus LD decays rapidly, i.e., at a rate similar to that observed in the outcrossing species, Zea mays (maize). Excess interlocus LD is observed at 15% of two-locus combinations; almost all interlocus LD involves loci with significant geographic structuring of mutational variation.

DNA, Plant↗
PubMed2004

Ageing populations and childhood infections: the potential impact on epidemic patterns and morbidity.

BACKGROUND: Population decline, arising from below replacement fertility and possibly giving rise to substantial changes in age distribution, is a feature of many industrialized developed countries; Italy is one of the most notable European examples. The potential influence of this phenomenon on prevalence of chronic non-infectious disease is well known, but little attention to date has been paid to the impact on severe disease due to childhood infections in those cases where control is insufficient to achieve elimination. METHODS: A transmission dynamics model incorporating realistic demography is used to investigate the possible impact of population decline and ageing and suboptimal vaccination uptake on the age distribution of incidence of measles infection and of consequent mortality. Data from Italy is used to parameterize the model. RESULTS: Population ageing in the absence of vaccination is shown to reduce per capita incidence of infection but also to increase average and upper quartile ages at infection. The effect is substantially enhanced by significantly suboptimal vaccination uptake, when disease-induced mortality has, for a period, the potential to exceed that in the absence of vaccination. CONCLUSIONS: Although a substantially increased burden from chronic non-infectious disease has frequently been proposed as a consequence of population decline, there is also potential for an increase in morbidity and mortality from measles and other childhood infectious diseases, particularly where vaccine uptake is substantially below the optimum. Rubella is highlighted as a particular cause for concern. This work also has implications for less-developed countries.

Adolescent↗
PubMed2005

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↗
PubMed2026

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↗