José Sulon

Pregnancies, calves and calf viability after transfer of in vitro produced bovine embryos.

Pregnancy, parturition and calf survival following the transfer of embryos produced in vitro were monitored. A total of 44 blastocysts was transferred in pairs to 1 uterine horn ipsilateral to the corpus luteum (CL) of 22 synchronized heifers. At Day 42 of development 14 recipients (64%) were pregnant; the calving rate was also 64%. The twinning rate was 9/14 at Day 42 and 7/14 at birth, for an overall fetal mortality rate of 9%. The average gestation length was 281 and 275 d for single and twin pregnancies, respectively. Blood samples from recipients were collected for determination of bovine pregnancy associated glycoprotein (bPAG) from 2 wk after transfer and throughout the pregnancy. During the first trimester of pregnancy, the bPAG concentration was significantly higher in twin than in single bearing heifers, and the perinatal increase in bPAG was correlated positively with the total weight of the fetus(es). The percentage of male calves was 43%. The birth weight of twin individuals was 25 +/- 1 kg, which was 78% of the birthweight of the singletons (32 +/- 2 kg). One singleton calf was oversized, weighing 58 kg (80% more than the median weight of the other singletons). Stillbirths occurred in 21% of the twins, butin none of the singletons. Calf mortality during the first 14 d was higher for twins (4/11) than for singletons (1/7) due to infections and cerebellar hypoplasia. Karyotyping the calves detected no cytogenetically recognizable abnormalities. All calves were negative for BVD virus and IBR antibodies. The results of this study showed that although the incidence of fetal loss was low, there was an unacceptable high perinatal mortality of the calves. Thus it is likely that the blood supply through the placenta of animals pregnant with twins was impaired or it is possible that these fetuses and calves had increased stress susceptibility caused by the in vitro conditions. Furthermore, the birth of 1 oversized calf, 2 calves with cerebellar hypoplasia and 5 calves succumbing to infections seems to indicate that a proportion of in vitro produced calves may suffer from factors inherent in the in vitro production system.

Early pregnancy diagnosis in goats by determination of pregnancy-associated glycoprotein concentrations in plasma samples.

Different RIA systems available for measuring the concentrations of pregnancy-associated glycoproteins (PAGs) in dairy goats were compared in order to evaluate their accuracy in early pregnancy diagnosis. Plasma concentrations of PAGs were determined by 3 heterologous RIA systems with a bovine PAG standard and tracer in combination with antisera anti-ovine PAG (RIA 1), anti-caprine PAG55 + 62 (RIA 2), anti-caprine PAG55 + 59 (RIA 3), and by 2 homologous RIA systems that employed caprine PAG55 + 62 and caprine PAG55 + 59 and their specific antisera (RIAs 4 and 5, respectively). In all of the RIAs, the mean concentrations of PAGs were significantly higher (P < 0.01) in pregnant than in nonpregnant goats from Day 21 onwards after breeding. On Day 21, the accuracy rates of early pregnancy diagnoses were 56% (RIA 1), 96% (RIA 2), 99% (RIA 3), 95% (RIA 4) and 90% (RIA 5), whereas on Day 28 these rates were > 99% for RIAs 2, 3, 4 and 5. The RIAs for PAGs depend on proteins from the placenta being present in maternal plasma and require only a single sample of blood, to distinguish pregnant goats from those that fail to return to estrus for other reasons. The homologous and semi-heterologous assays are highly accurate as early as Day 21 of pregnancy.

Effect of the presence of females on the pituitary-testicular activity in male Japanese quail (Coturnix coturnix japonica).

Five experiments were carried out to study the role of the presence of a female on the reproductive endocrinology of male Japanese quail. In the first three experiments, exposure of an adult male raised in long days to a female for l0 min or l week failed to increase plasma testosterone and LH levels; in fact a significant transitory decrease in plasma testosterone was observed, associated with a preceding increase in plasma corticosterone. These changes are interpreted as a result of the stress caused by repeated bleeding or by the continuous presence of a female in a limited space. In the last two experiments, an increase in the maturation rate of immature males could be observed in birds maintained in the continuous presence of females by comparison with birds kept in isolation. The paired males had larger cloacal glands and testes and higher plasma levels of testosterone and LH than the isolated one. This effect of the female was observed in long days (l6L:8D) as well as in marginally stimulating short days (l2L:l2D).

Testosterone metabolism and testosterone-dependent characteristics in Japanese quail.

In 2 independent experiments, we measured and correlated in maturing male Japanese quail the individual variations in sexual and aggressive behavior, cloacal gland size, testes weight, plasma testosterone concentrations and intracellular testosterone metabolism by hypothalamus and cloacal gland. Cloacal gland area was only weakly related to plasma testosterone levels but was positively correlated with the production of active androgenic metabolites and negatively related to the production of 5 beta-reduced androgens (inactive) in the glandular tissue. Several measures of behavior were correlated with aspects of the testosterone metabolism in the anterior hypothalamus. In both experiments, the behavior of the birds was also strongly correlated with their testes weight and their cloacal gland area but weakly or not at all with their plasma testosterone levels. These studies suggest that testosterone metabolism is involved in the control of hormone action in maturing animals.

Early pregnancy diagnosis in sheep by progesterone and pregnancy-associated glycoprotein tests

The aim of this study was to compare the accuracy of the progesterone (P4) and pregnancy associated glycoprotein (PAG) tests for determination of early pregnancy in sheep. Estrus was synchronized in 182 Awassi x Merino ewes and blood samples were collected at Days 0 (day of the insemination), 18, 22, 29, 36, and 50 after artificial insemination (AI). Plasma P4 concentrations at Days 0 and 18 were determined by double antibody radioimmunoassay, while PAG concentrations at Days 22, 29, 36 and 50 were determined by a heterologous, double-antibody radioimmunoassay (RIA) using the bovine PAG 67 kDa subunit as tracer and standard and rabbit antiserum raised against a mixture of caprine 55 and 59 kDa PAG subunits as the first antibody. The discriminatory value for diagnosis of pregnancy by the P4 and the PAG-RIA tests was > or = 1 ng/ml. Based on lambing data, the accuracy for diagnosing pregnant (sensitivity) and non-pregnant ewes (specificity) and predictivity of both tests were calculated. The sensitivity, specificity, positive and negative predictive values for P4 and PAG tests were 100, 95.4, 81.6, and 100% at Day 18 (P4) and 93.5, 100, 100 and 98.7% at Day 22 (PAG), respectively. For diagnosis of non-pregnant ewes the PAG test had significantly higher specificity than the P4 test (P < 0.05). It is concluded that ovine pregnancy can be reliably diagnosed at Day 22 after AI by using a heterologous radioimmunoassay of PAG.

Hypothalamic-pituitary-adrenal reactivity in prepubertal children with social phobia

BACKGROUND The aim of this study was to investigate whether a different pattern of HPA axis activity is found between children with social phobia (SP) and healthy control children. METHODS A total of 50 prepubertal subjects (25 children with SP and 25 healthy control subjects) were studied. The effects of stress were studied by comparing cortisol responses to a psychosocial stressor, consisting of a public speaking task. RESULTS Children with SP showed an elevated cortisol response to the psychosocial stressor as compared with healthy controls. Trait but not state anxiety levels are associated with higher HPA axis activity. LIMITATIONS Limited sample size. CONCLUSIONS The results indicate that a higher cortisol responsivity to stress may be a neurobiological marker for prepubertal children with SP. Directions for future research and clinical implications are discussed.

Modulatory effect of imetit, a histamine H3 receptor agonist, on C-fibers, cholinergic fibers and mast cells in rabbit lungs in vitro

The pharmacological mechanisms involved in the interactions between C-fibers, cholinergic fibers and mast cells were investigated in tracheally perfused rabbit lungs by measuring the simultaneous release of substance P and histamine in lung effluents. The amounts of substance P and histamine released in lung superfusates were measured by radioimmunoassay (RIA) after administration of capsaicin and carbachol. Capsaicin (10(-4) M) induced a simultaneous increase in substance P (273 +/- 56% of baseline) and histamine (460 +/- 138%) release. Similarly, carbachol (10(-4) M) caused an increase in the release of both substance P (367 +/- 111%) and histamine (1379 +/- 351%). The effect of capsaicin was prevented by pretreating the lungs with the tachykinin NK1 receptor antagonist SR 140333 (10(-7) M), and atropine (10(-6) M). SR 140333 prevented the carbachol-induced release of substance P but not of histamine. Exogenous substance P induced an increase in histamine release (136 +/- 7%) which was significantly greater in lungs perfused with the neutral endopeptidase inhibitor, thiorphan (10(-5) M) (272 +/- 35%). This effect was prevented by atropine (10(-6) M). Pretreatment of lungs with imetit (5 x 10(-8) M), a selective H3 receptor agonist, prevented the capsaicin-induced release of both mediators. Imetit also blocked the carbachol-induced release of substance P but not of histamine. Exogenous substance P-evoked histamine release was inhibited by imetit. Therefore, it can be concluded that substance P released through the action of capsaicin can activate cholinergic fibers, leading to cholinoceptor stimulation with subsequent activation of C-fibers and mast cells. While the presence of presynaptic H3 receptors modulating substance P-induced acetylcholine release was only surmised, the existence of modulating histamine H3 receptors on C-fibers was confirmed.

Use of saliva cortisol in the dexamethasone suppression test

In a sample of 26 inpatients (15 primary endogenous depressives and a heterogeneous comparison group of 11 psychiatric patients), results of the dexamethasone suppression test (DST) for endogenous depression were compared when cortisol was measured in plasma (total and free) and in saliva. Results showed a close linear relationship among plasma total and free cortisol, plasma total cortisol, and saliva cortisol, and between free plasma and saliva cortisol. A saliva cortisol cutoff point of 70 ng/dl achieved the same sensitivity (67%), specificity (91%), and diagnostic confidence (91%) as the best cutoff scores of plasma total cortisol (5 micrograms/dl) and plasma free cortisol (0.15 microgram/dl). These results suggest that saliva cortisol, which directly reflects the biologically active fraction of cortisol, can be used as a reliable and more practical index in the DST, especially in outpatients.

Diurnal variations of sexual receptivity in the female Japanese quail (Coturnix coturnix japonica).

A series of experiments was performed to study the changes during the day of female receptivity in Japanese quail. In each experiment, the frequency of squatting and the percentage of male approaches which were followed by squatting increased at the end of the day, approximately 11 to 13 hr after lights on (in a photoperiod of 16L:8D). In some cases this increased receptivity was associated with a significant decrease of the long-avoid frequency. Analyses are presented which demonstrate that the increased receptivity at the end of the day is not directly caused by the oviposition and does not result directly from changes in the behavior of the male stimuli. This is strongly supported by the observation that the increase in receptivity was observed at the same time after lights on (but different clock times) in two groups of females which were raised in two different photoperiods shifted by 6 hr and tested with the same group of males raised in one of the two photoperiods. The increase in receptivity coincides with an increase in plasma estradiol and progesterone. Considering that this behavior is suppressed by ovariectomy, it is argued that the daily changes in receptivity could be controlled by the hormonal changes associated with the ovulatory cycle.

Exteroceptive Silent Period of Temporalis Muscle in Menstrual Headaches

The second exteroceptive silent period (ES2) of temporalis muscle was recorded on days 1 and 15 of the menstrual cycle in 17 women, 9 of whom suffered regularly from tension-type headaches during menstruation. Mean duration of temporalis ES2 was significantly shorter on day I of the cycle than on day 15. This difference was due to a marked menstrual reduction of ES2 in the headache subgroup. A positive correlation was found between ES2 durations and oestradiol/progesterone ratios. We hypothesize that the variations of ES2 during the ovarian cycle result from the modulatory effects of oestrogens on descending aminergic pathways that control excitability of inhibitory brainstem interneurons mediating exteroceptive suppression of jaw-closing motoneurons.