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Distribution of antibodies to porcine circovirus in swine populations of different breeding farms.

An enzyme linked immunosorbent assay (ELISA) was developed for mass antibody screening to porcine circovirus (PCV) in pig herds of different age groups and of different husbandries. Infection with PCV was found to be common in all swine herds tested, with only one exception, a herd at a small farm. Statistically, the percentage of PCV negative sera decreased and titer levels increased with increasing age of the pigs. Within individual age groups, differences were found to exist between different husbandries. No correlation was detected between antibody levels and reproductive disorders in the herds.

Age Factors↗

Diagnosis and treatment of acromegaly complications.

The Pituitary Society in conjunction with the European Neuroendocrine Association held a consensus workshop to develop guidelines for diagnosis and treatment of the co-morbid complications of acromegaly. Fifty nine pituitary specialists (endocrinologists, neurosurgeons and cardiologists) assessed the current published literature on acromegaly complications in light of recent advances in maintaining tight therapeutic control of GH hypersecretion. The impact of elevated GH levels on cardiovascular disease, hypertension, diabetes, sleep apnea, colon polyps, bone disease, reproductive disorders, and neuropsychologic complications were considered. Guidelines are proposed for effective management of these complications in the context of overall acromegaly control. When appropriate, requirements for prospective evidence-based studies and surveillance database development are enunciated. Effective management of co-morbid acromegaly complications will lead to improved morbidity and mortality in acromegaly.

Acromegaly↗

The first finding of a Rickettsia bacterium associated with parthenogenesis induction among insects.

The larval endoparasitoid, Neochrysocharis formosa (Westwood), is an important natural enemy of the leafminer Liriomyza trifolii (Burgess) in Japan. The thelytokous strain of N. formosa mostly produces female progeny. Male progeny were produced by females treated with tetracycline, suggesting that microorganisms induce thelytoky in N. formosa. The result of nucleotide sequencing of the 16S rRNA gene indicated that the parasitoid is infected with a Rickettsia bacterium, which appears to be causative of the thelytoky. Although Rickettsia-bellii-like bacteria have been found to be associated with various reproductive disorders, this is the first finding of a parthenogenesis-inducing Rickettsia among insects.

Animals↗

Organotypic culture, a powerful model for studying rat and mouse fetal testis development.

The key role of the fetal testis in the masculinization of genital organs has been known for a long time. More recently, the observed increases in male reproductive disorders has been postulated to be the result of changes in fetal and neonatal testis development in response to increasing environmental pollution. However, few tools are available for studying fetal testis development and the effects of physiological or toxic substances. We have developed an organ culture system in which rat fetal testis is grown on a filter floating on a synthetic medium containing no serum, hormones or biological factors. In this study, we have compared the long-term morpho-functional development of the various testicular cell types in this system with that observed in vivo and have extended this system to the mouse. Rat Leydig, Sertoli and germ cells and macrophages develop normally over a period of 1-2 weeks in this system. Fewer cells are produced than in vivo but the level of differentiated function is similar. Germ cells, which are difficult to culture in vitro, resume mitosis after a quiescent period, at the same time as in vivo. Similar results have been obtained with mouse fetuses, except that Leydig cells dedifferentiate in vitro if the testis is explanted after 13.5 days post conception. Testicular architecture and intercellular communication are sufficiently preserved for the development of the main fetal and neonatal testicular cell types in vitro with no added factors. Our floating-filter organotypic culture system in synthetic medium therefore allows the morpho-functional development of somatic and germ cells in fetal testis explants taken at all developmental stages in rat and at early stages in mouse. This method is potentially useful for studies of the effects of various factors, and of xenobiotics, in particular.

Animals↗

Vascular morphogenesis in the primate ovary.

Ovarian function is dependent on intense cyclical vascular morphogenesis and regression. The molecular and cellular pathways involved in the generation of new capillary networks in the ovary are now being elucidated. Focussing on the marmoset, the course of angiogenesis at different stages of follicular maturation and in the corpus luteum throughout the cycle and in early pregnancy have been quantified and major progress has been made in the evaluation of the role of vascular endothelial growth factor (VEGF). To study the physiological role of VEGF in follicular and luteal angiogenesis in detail, VEGF was inhibited during defined stages of the cycle in vivo. VEGF antagonist administered throughout the follicular phase of the cycle resulted in a marked decrease in endothelial cell proliferation in developing antral follicles, accompanied by a decline in granulosa cell proliferation, restriction of follicular growth and inhibition of ovulation. An outstanding feature in the ovary is the intense angiogenesis that occurs during the early luteal phase. VEGF inhibitors markedly suppressed this angiogenesis, resulting in a marked restriction in the development of the microvascular tree and suppression of plasma progesterone. These studies showed that VEGF is essential for normal follicular and luteal angiogenesis and function, and demonstrated how luteal angiogenesis in particular could serve as a sensitive bioassay for putative angiogenic antagonists. Antagonists of VEGF are potent tools for investigating the role of angiogenic factors within the ovary and may have applications to the treatment of reproductive disorders characterised by alterations in normal vascular structure or function.

Angiogenesis Inducing Agents↗

Predicting human cloning acceptability: a national Greek survey on the beliefs of the public.

BACKGROUND: In the era of human genome research, there is a large theoretical debate among scientists and authorities on the ethical dimension based on the moral liberty of the individuals and the scientific and economic dimension based on the freedom and the independence of the scientific and technological activities. Meanwhile, the understanding of beliefs on human cloning (HC) and its acceptability are important for the development of evidence-based policy making. However, previous research in the field of public beliefs towards human genetics is limited. Therefore, in the present study, we investigated the role of public beliefs as predictors of the acceptability of the respondents to use HC. METHODS: Personal interviews were conducted with 1020 men and women of urban areas in Greece. Stratified random sampling was performed to select participants. Several scientists, experts in HC, evaluated the content of the instrument initially developed. The final questionnaire was the result of a pilot study. RESULTS: The acceptability of HC for the cure of incurable diseases and transplantation need is very high (70.7 and 58.6%, respectively). Public's intention to have recourse to HC because of "bringing" back to life a loved person or because of reproductive disorders was reported by 35 and 32.5%, respectively. With respect to the role of beliefs: increasing scores of reasons of social benefits, moral/religious reasons and legislative reasons increased the public's intention to have recourse to HC; inversely, decreasing scores of reasons of human commodification/exploitation increased public's intention to have recourse to HC. Additionally, low rates of church attendance appeared to be correlated with high reported acceptability of HC. CONCLUSION: There is great public concern regarding the application of HC, which probably reflects the existing ambivalence over the relationship between technology and society. Scientists and policymakers should take into account these indicators of public disquiet and should manage the public involvement in policy decisions, from which they have so far been excluded.

Adolescent↗

High prevalence of chlamydial (Chlamydophila psittaci) infection in fetal membranes of aborted equine fetuses.

Seventy-seven cases of equine abortion from 49 Hungarian farms that occurred between 1998 and 2000 were investigated for the presence of chlamydiae by immunohistochemistry, PCR and/or MZN staining. Evidence of the presence of these bacteria was obtained in 64 cases (83.1%) from 41 (83.7%) different farms. Partial ompA gene sequencing of PCR products revealed that the agent was Chlamydophila psittaci. Based on the findings of microbial diagnosis, pathology and case history, chlamydial infection was considered to be the most likely cause of abortion in at least 11 (14.3%) cases. In the remaining 53 Chlamydophila-positive cases, either other bacterial and viral agents (n = 22 or 28.6%) as well as non-infectious factors (n = 14 or 18.2%) were identified as more probable primary causes of disease, or the role of chlamydiae remained unclear because lesions in fetuses and fetal membranes were absent (n = 17 or 22.1%). When chlamydial antigen was detected in aborted equine placental tissue using immunohistochemistry it was seen only in the chorionic epithelial cells, but not in other parts of the fetal membranes nor in any of the fetal tissues. In conclusion, chlamydial infection of the genital tract should be considered a possible factor in equine reproductive disorders.

Aborted Fetus↗

A clinical evaluation of a direct radioimmunoassay of testosterone.

We report the evaluation of a commercial coated tube kit (Medgenix) to the measurement of testosterone in plasma/serum samples. The accuracy, precision, specificity and recovery of the assay and the affinity of antibody for ligand were established. The range of concentrations of testosterone providing satisfactory analyses was 0.48-48 nmol/l, which is appropriate for general clinical use. Results of analysis of male and female samples obtained from the routine hospital laboratory in endocrinologically-defined groups were also reviewed. In all groups of male subjects and in normal females, the kit provided results in close agreement to those of an extraction method. With samples from women with various reproductive disorders, as well as with some samples obtained from external quality controls, the kit showed a small but significant negative bias relative to the results of the extraction method. There was, however, no evidence of bias with samples from a separate group of volunteer women with the polycystic ovary syndrome. No interference due to SHBG was detected. Clinical sensitivities using the kit were equivalent to those obtained using the extraction RIA. There was no evidence of spuriously high results in samples from females which has been a common problem with direct radioimmunoassay of testosterone.

Adolescent↗

Direct enzymeimmunoassay of progesterone in bovine milk.

A sensitive enzymeimmunoassay has been developed for measuring progesterone in unextracted bovine milk. An N-hydroxysuccinimide ester of 11 alpha -hydroxyprogesterone 11-hemisuccinate has been synthesised and used to form conjugates with beta-galactosidase in buffer at pH 7.0. The degree of incorporation of progesterone into the enzyme was demonstrated using (14C)-labelled steroid and by radioimmunoassay binding inhibition. Standard curves of comparable range and sensitivity to radioimmunoassay were obtained in the presence of whole milk taken from a cow at oestrus. These advances have allowed the development of a simple micro-titre plate enzymeimmunoassay of progesterone in whole milk and will be of particular value in determination of pregnancy, prediction of the day of oestrus and diagnosis of reproductive disorders.

Animals↗

The effects of prolactin on rat ovarian function.

Hyperprolactinemia has been associated with several reproductive disorders. To investigate whether hyperprolactinemia directly affects rat ovarian function, we examined the ovarian histopathology and the activities of the four ovarian enzymes 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 17-hydroxylase (17-OH), 17,20-desmolase (17,20-D) and aromatase in hyperprolactinemic rats and controls. Hypophysectomized, gonadotropin-treated Fisher rats were made hyperprolactinemic by isografting pituitary glands under the kidney capsule. The control animals received skeletal muscle. The ovaries were resected, pooled according to prolactin levels and microsomal enzyme activities were measured from each pool. Prolactin (PRL) levels were 344 +/- 23 ng/ml in the hyperprolactinemic rats and 18 +/- 5 ng/ml in the controls (p less than 0.001). Estradiol concentrations were 609 +/- 47 pg/ml in the hyperprolactinemic animals and 56 +/- 13 pg/ml in the controls (p less than 0.001). Ovarian and uterine weights were significantly higher in the hyperprolactinemic rats (p less than 0.02). Ovarian histopathology demonstrated benign polycystic transformation in the hyperprolactinemic animals. Hyperprolactinemia had no effect on 3 beta-HSD, but was associated with significant decreases in the 17-OH, 17,20-D and aromatase activities when compared to controls (p less than 0.001). We conclude that prolactin has a direct effect on rat ovarian function which appears to be independent of changes in gonadotropin secretion.

3-Hydroxysteroid Dehydrogenases↗

Effect of PG F2 alpha treatment on conception rates of dairy cows treated with a GnRH agonist 12 to 14 days after Artificial Insemination.

The effect of a GnRH agonist (10 ug Buserelin) on conception rate was determined when injected into dairy cows 12 to 14 days after Artificial Insemination (AI). The following factors were taken into account: previous treatment prior to AI with Prostaglandin F2 alpha, clinical history recorded prior to AI since last calving, parity and milk yield. A number of 118 cows, from one large dairy herd, were involved in this study. A total of 210 AI's were performed, followed by 140 GnRH treatments and 70 saline injections. Reproductive events were then recorded. The rank of AI was equally distributed among the two groups. 153 AI's were preceded by a Prostaglandin F2 alpha (Dinoprost - 25 mg) treatment among which 103 were subsequently GnRH-treated and 50 were controls. Plasma progesterone concentrations were determined daily for 34 days after AI in 13 treated and 13 control cows. An Early Pregnoncy Diagnosis (EPD) from milk progesterone concentration was performed on Day 21 after AI in all cases. Post-AI GnRH agonist treatment resulted in a significant enhancement of the conception rate: 60% vs 44%; p < 0.01, respectively for treated and control animals. PG F2 alpha treatment prior to GnRH injection had a major influence on the conception rate (62% vs 40%; p < 0.01). No effect was seen (54 % vs 55 %) in non-PG F2 alpha treated females which were subsequently injected respectively with GnRH or saline after AI. Previous clinical reproductive disorders, parity and milk yield had no significant effect. In non-pregnant treated animals, GnRH agonist treatment resulted in an increased rate of heat detection on days 20 - 25 after AI (91 % vs 74 %) and a higher fertility rate on the following AI was seen (59 % vs 44 %; p < 0.05). In conclusion, GnRH agonist treatment 12 to 14 days after AI only enhanced the conception rate of females which had previously been treated with PG F2 alpha. In non-pregnant cows, this treatment had also a benficial effect on heat detection and improved the conception rate at the subsequent AI.

Journal Article↗

Follicular development and reproductive endocrinology during and after superovulation in heifers and mature cows displaying contrasting superovulatory responses.

To understand the causes for poor response to superovulation in mature cows of high genetic potential, endocrine and follicular events during and after superovulation were compared in heifers (<2 yr old) yielding large numbers of embryos and cows (9 to 13 yr old) known to be poor embryo donors. Follicular development was monitored by daily ultrasonography. Blood samples were taken 2 to 3 times a day for the measurements of P4, E2, FSH and LH by RIA. Intensive blood collections at 15-min intervals for 6 h were also performed during preovulatory and luteal phases. The number of embryos produced in the heifers (15.2 +/- 2; mean +/- SEM) and the cows (0.6 +/- 0.4), was similar to the number of ovulatory follicles derived from ultrasonographic observations in the heifers (16.2 +/- 3.7), but not in the cows (7.8 +/- 2.8). Contrary to that observations in heifers, there was no increase in the number of 4- to 5-mm follicles in cows during superovulation. The number of larger follicles (>5 mm) increased during superovulation in both cattle groups, but it was significantly lower in cows than in heifers. During superovulation, the maximal E2 concentration was greater (P < 0.0001) in heifers than in cows. One cow showed delayed luteolysis during superovulation, while another had abnormally high FSH (>10 ng/ml) and LH (>3 ng/ml) concentrations following superovulation. All the cows had a postovulatory FSH rise which was not detected in the heifers. The results showed that attempts to improve superovulatory response in mature genetically valuable cows are hampered by a number of reproductive disorders that are not predictable from the study of the unstimulated cycle.

Journal Article↗

Effect of lindane on mitochondrial side-chain cleavage of cholesterol in mice.

Cholesterol side-chain cleavage of mitochondria, the key rate limiting step in steroid biosynthesis in ovarian tissues, was studied in female Swiss mice fed lindane (gamma-hexachlorocyclohexane) at various doses and over varying periods. The insecticide adversely affected cholesterol side-chain cleavage of the ovary as judged by decreased conversion of this sterol to pregnenolone and subsequently to progesterone in a dose-dependent manner. The formation of pregnenolone was maximally inhibited (75% inhibition) at the highest intake of insecticide with simultaneous inhibition of its conversion to progesterone. At all doses, the rate of inhibition of pregnenolone formation was comparatively higher than its conversion to progesterone. The significant inhibition of cholesterol side-chain cleavage by lindane and resultant decrease in the rate of steroidogenesis in the ovary would account for the observed gonadal hormone deficiency and related reproductive disorders in the lindane-fed mice.

Administration, Oral↗

Pathogenesis of porcine circovirus; experimental infections of colostrum deprived piglets and examination of pig foetal material.

The results of virus and antigen distribution following experimental infection of colostrum deprived pigs with pig circovirus (PCV) by oral/nasal and intravenous routes are reported. PCV and antigen were detected using virus isolation and indirect immunofluorescence on cryostat sections respectively. PCV antigen was detected in tissues throughout the body but primarily in spleen thymus, and lung. No PCV antigen or virus was detected in tissue samples from the central nervous system. Examination of pig foetal material from field cases of abortion/stillbirth resulted in 3 PCV isolates from 2 sera and a spleen sample from 2 groups of stillborn piglets from the same farm. No antibody to PCV alone was detected in 160 foetal sera tested. These results suggest that transplacental infection with PCV does occur, possibly prior to foetal immunocompetance. However, it is probably not a significant cause of reproductive disorders in pigs in Northern Ireland.

Animals↗

Isolation of porcine reproductive and respiratory syndrome (PRRS) virus in a Danish swine herd and experimental infection of pregnant gilts with the virus.

The first case of porcine reproductive and respiratory syndrome (PRRS) in Denmark was diagnosed in March 1992 by the detection of specific antibodies against PRRS virus in serum samples originating from sows in a herd located on the island of Als. Subsequently, PRRS virus was isolated from a 200-sow farrow-to-finish herd with clinical signs consistent with PRRS. The virus was isolated by inoculation of pleural fluid from a stillborn piglet onto porcine pulmonary alveolar macrophages. The isolate was identified as PRRS virus by staining with a specific antiserum. By electron microscopy, the virus particle was found to be spherical and enveloped, measuring 45-55 nm in diameter and containing a 30-35 nm nucleocapsid. Only minor antigenic differences were found between the Danish and a Dutch isolate. Following intranasal inoculation of 3 pregnant gilts with the Danish isolate transplacental infection was demonstrated by the re-isolation of PRRS virus from approximately 45% of the piglets from the experimentally infected gilts. However, the experimental infection produced no significant reproductive disorders or other clinical signs. At autopsy, histopathological examination revealed slight interstitial pneumonia in a few piglets.

Abortion, Veterinary↗

Spontaneous recovery from ethanol-induced male infertility.

Chronic ethanol ingestion by the male is associated with reproductive impairment. No longitudinal studies have been carried out to determine the recovery of ethanol-related reproductive failure subsequent to moderate periods of abstinence. An animal model (C57B1 mouse) was utilized to examine the effectiveness of abstinence for reversal of ethanol-induced reproductive failure. After treatment with either a 5% ethanol diet (10 weeks) or a 6% ethanol diet (5 weeks), animals were hemicastrated (right testis and accessory organs); their reproductive tracts and epididymal spermatozoa were examined. Ingestion of the 5% and 6% ethanol diets resulted in significantly (p less than 0.05) decreased testicular weights (24% and 28%, respectively) and seminal vesicle/prostate weights (20%, 6% diet only), increased frequencies of germ cell desquamation (480% and 400%), inactive seminiferous tubules (186% and 567%) and inhibition of in vitro fertilization of mouse oocytes by epididymal spermatozoa (26% and 62%), as compared to their respective pair-fed control values. Also observed was a significant decrease in total motile spermatozoa (85%, 6% diet only). Improvement in all parameters was observed in the contralateral reproductive organs subsequent to ten weeks abstinence. Only germ cell desquamation remained significantly elevated (100% as compared to control) in animals that ingested the 5% diet. In contrast, significant abnormalities persisted ten weeks after treatment with the 6% ethanol diet, including increased germ cell desquamation (200%) and inactive seminiferous tubules (157%); decreased forward progression (17%) of epididymal sperm also persisted. If these data can be applied clinically, male alcoholic patients with reproductive disorders could have a prognosis of at least partial recovery following moderate periods of abstinence from ethanol.

Animals↗

The variability in the interval between estrus and ovulation in cattle and its determinants.

Fertility of Holstein cows has been decreasing for years and, to a lesser extent, the fertility of heifers too but more recently. A hypothesis to explain this phenomenon may be that the chronology of events leading to ovulation is different for those animals bred nowadays when compared to what was reported previously; this would result in an inappropriate time of insemination. Therefore, two experiments were designed to investigate the relationships among estrus behavior, follicular growth, hormonal events and time of ovulation in Holstein cows and heifers. In the first experiment, the onset of estrus, follicular growth, patterns of estradiol-17beta, progesterone and LH, and the time of ovulation were studied in 12 cyclic Holstein heifers that had their estrus synchronized using the Crestar method; this was done twice, 3 weeks apart. The intervals between estrus and ovulation, estrus and the LH peak, and between the LH peak and ovulation were, respectively, 38.5 h +/-3.0, 9.1 +/- 2.0 and 29.4 h +/-1.5 (mean+/- S.E.M). The variation in the interval between estrus and the LH peak explained 80.6% of the variation in the interval between estrus and ovulation. The intervals between estrus and the LH peak, and estrus and ovulation were correlated with estradiol-17beta peak value (r=-0.423, P <0.04 and r=-0.467, P<0.02, respectively). Positive correlation coefficients for the number of follicle larger than 5 mm, and negative correlation coefficients for the size of the preovulatory follicle with the intervals between estrus and LH peak, LH peak and ovulation, and estrus and ovulation suggest an ovarian control of these intervals. In respect to its role to explain the variation in the interval between estrus and ovulation, the variation in the interval between estrus and the LH peak was evaluated further in a second set of experiments utilizing 12 pubertal Holstein heifers and 35 Holstein cows. The duration of the interval between the beginning of estrus and the LH peak was longer in heifers than in cows (4.15 h versus -1.0 h; P <0.002); the variation for this interval was higher in cows than in heifers (S.E.M.= 1.2 h versus 0.8 h; P=0.01). According to the results of these studies it can be proposed that estradiol and other product(s) of ovarian origin regulate not only the duration of intervals between the onset of estrus and the LH surge but also between the LH surge and ovulation. From the results obtained in the first experiment, it may be postulated that differences observed between cows and heifers for the duration of the interval between onset of estrus and the LH surge as well as for the variation of this interval would be observed also for the interval between the onset of estrus and ovulation. Therefore, on a practical point of view, the long interval between the onset of estrus and ovulation and the high variation of this interval, especially in cows, may be a source of low fertility and should be considered when analysing reproductive disorders.

Animals↗

The uterine junctional zone.

Magnetic resonance imaging has revealed that the endometrio-myometrial interface constitutes a distinct, hormone-dependent uterine compartment termed the junctional zone. In the non-pregnant uterus, highly specialized contraction waves originate exclusively from the junctional zone and participate in the regulation of diverse reproductive events, such as sperm transport, embryo implantation, and menstrual shedding. Conversely, growing evidence suggests that disruption of the normal endometrio-myometrial interface plays an integral role in diverse reproductive disorders. This chapter reviews our current understanding of the mechanisms that govern the cyclic changes in the uterine junctional zone and summarizes the evidence implicating the endometrio-myometrial interface in normal uterine physiology and pathological processes.

Endometriosis↗