[Characteristics of jugular venous pulse and its genesis in Ebstein's anomaly]. To clarify the characteristics of the jugular venous pulse and its genesis in Ebstein's anomaly, 6 patients with Ebstein's anomaly whose mean age was 45 +/- 9 years, and 10 normal subjects with a mean age of 27 +/- 6 years were studied by phono-mechanocardiography and echocardiography. The parameters included the relative height of the jugular "c" wave, the relative timing of the upstroke (Q-Cu interval) and the peak of the jugular "c" wave (Q-Cpeak interval), the relative timing of the upstroke of the carotid artery pulse, the relative timing of tricuspid valve closure (Q-Tc interval), the excursion of the closing motion of the anterior tricuspid leaflet (TV excursion), the distance between the anterior mitral annulus and the septal tricuspid annulus (M-T distance), the area of atrialized right ventricle (ARV area) and the maximum area of the tricuspid regurgitant signal. Among 6 patients with Ebstein's anomaly, tricuspid regurgitation was predominant in 4 and mild in the remaining 2. The results were as follows: 1. A large jugular "c" wave was observed in 4 of the 6 patients. Two patients with large ARV area had giant "c" wave. 2. The interval of the upstroke of the carotid artery pulse and that of the jugular "c" wave was about 39.2 msec. 3. The Q-Tc interval was significantly longer and the TV excursion was significantly greater in the patients than in the normal controls. 4. There was only a weak positive correlation between the Q-Cpeak interval and the Q-Tc interval. No significant correlation was observed between the relative height of the jugular "c" wave and the TV excursion. 5. There was a positive correlation between the relative height of the jugular "c" wave, the ARV area and M-T distance. 6. No obvious correlation was observed between the grade of tricuspid regurgitation and the relative height of the jugular "c" wave. These results suggest that augmentation of the "c" wave of the jugular venous pulse is characteristic of Ebstein's anomaly and that it correlates closely with the severity of displacement of the tricuspid valve and the size of the atrialized right ventricle.


[Pulmonary uptake of thallium-201 in patients with congenital heart disease: comparison between total anomalous pulmonary venous connection and tetralogy of Fallot]. To evaluate the pulmonary extravascular space in patients with congenital heart disease, lung uptake of thallium-201 (T1-201) was quantitatively studied. A total of 50 T1-201 imagings were performed in 33 patients with total anomalous pulmonary venous connection (TAPVC); 4 preoperatively, 22 postoperatively in the early stage (within 6 months), and 24 in the late stage (7 months or later). The images consisted of 17 supracardiac TAPVC (type-I), 13 paracardiac (type-II) and 3 infracardiac (type-III). In patients with tetralogy of Fallot (T/F), T1-201 imaging was performed 15 times preoperatively, 12 in the early stage and 15 in the late stage, postoperatively. Furthermore, 29 patients with ventricular septal defect (VSD) or patent ductus arteriosus (PDA) were also studied preoperatively, and 21 in the late postoperative stage. Twenty-five patients with arrhythmias or a history of Kawasaki disease without perfusion defects were studied on T1-201 myocardial imaging. Lung uptake of T1-201 was analyzed with a computer using the anterior image of the chest, and the average count ratio of the right lung (P) to the left ventricular wall (LV) was calculated. P/LV values were compared between the patients before and after surgery, and differences in anatomical types in TAPVC were also evaluated. In TAPVC, P/LV values decreased gradually in the postoperative state, but were significantly higher than those of controls even in the late stage. In the late postoperative stage, type-I TAPVC had significantly higher P/LV values than those of type-II.(ABSTRACT TRUNCATED AT 250 WORDS)


[Echocardiographic assessment of aortic regurgitation and aortic root dilatation in bicuspid aortic valve]. Aortic regurgitation (AR) and aortic root dilatation in 29 consecutive patients with bicuspid aortic valves but without aortic root disease (20 males, and 9 females: aged 27-85 years) were studied using two-dimensional echocardiography. The normal ranges of aortic root dimensions were calculated from values of 185 normal subjects, as 95% confidence intervals. AR was observed in 17 patients by color flow mapping. In 12 of the 17 AR patients, no significant lesion of the aortic cusp was detected by two-dimensional echocardiography. These 12 AR patients were compared with 12 patients without AR. Increase in dimension of the aortic root was relatively frequent in the 12 AR patients at the aortic annulus (AA) (67 vs 17%, p < 0.05), and at the sinus of Valsalva (A1) (67 vs 17%, p < 0.05). At the ascending aorta 5 mm distal to the sinus of Valsalva (A2), the difference was not significant (58 vs 17%, p < 0.09). The 12 bicuspid AR patients without significant lesions of the aortic cusp were compared with 41 AR patients with normal tricuspid aortic valves. The frequencies of cases with increased aortic root dimension were 67 vs 46% (ns) at the AA, 67 vs 22% (p < 0.05) at A1 and 58 vs 5% at A2 (p < 0.01). Thus, aortic annular dilatation was thought to be the cause of AR in bicuspid and tricuspid aortic valves without significant lesions of the aortic cusps, and generalized dilatation of the aortic root was more frequent in bicuspid AR patients than in tricuspid AR patients.(ABSTRACT TRUNCATED AT 250 WORDS)


[Effects of chronotropic responsive cardiac pacing on ventilatory response to exercise in patients with bradycardia]. To identify the effect of chronotropic responsive cardiac pacing on ventilatory responses to exercise, 9 patients with chronotropic incompetence underwent paired cardiopulmonary exercise tests with fixed demand rates (AAI, VVI) and chronotropic responsive (AAIR, VVIR, DDD) pacing modes. Compared with fixed rate pacing, chronotropic responsive pacing increased peak oxygen uptake and delayed the attainment of the anaerobic threshold (AT) with a higher level of oxygen consumption (p < 0.01). Dyspnea was a major symptom that limited exercise time in 7 patients with fixed rate pacing, which was prominent with chronotropic responsive pacing. Ventilation (VE) and the ratio of ventilation to CO2 production (VE/VCO2) were consistently higher with fixed rate pacing during exercise. To compare the responses between the 2 pacing modes with the same work loads under aerobic conditions, we measured ventilatory variables one min prior to the AT as obtained with fixed rate pacing. When switching the pacing mode from fixed rate pacing to chronotropic responsive pacing, VE and VE/VCO2 decreased significantly from 22.0 +/- 7.8 to 19.8 +/- 6.8 l/min, and from 37.4 +/- 5.4 to 33.6 +/- 5.2, respectively. Tidal volume did not change, but respiratory frequency decreased more with chronotropic responsive pacing (p < 0.05). Although peak VE did not differ between the 2 pacing modes, VE/VCO2 decreased more with chronotropic responsive pacing (p < 0.01). Respiratory frequency decreased and tidal volume increased more with chronotropic responsive pacing (p < 0.05). This study suggests that chronotropic responsive cardiac pacing attenuates exertional dyspnea by improving ventilatory responses to exercise as well as increasing the cardiac output in patients with chronotropic incompetence.


[The influence of atrioventricular asynchronous contraction on left and right ventricular performance]. To investigate the influence of atrioventricular asynchronous contraction on left and right ventricular performance, pulsed Doppler echocardiographic studies were performed in 10 patients who received permanent pacemaker (VVI mode), but without significant heart disease except for complete heart block. After setting the pacing rate at 40 per min, the performance was analyzed during the patient's own slow ventricular rate. Flow velocity patterns at the left (LVOT) and right ventricular outflow tracts (RVOT) were recorded by pulsed Doppler echocardiography, and ejection time (EjT), acceleration time (AcT), peak velocity (PV) and flow velocity integral (FVI), which is proportional to stroke volume, were measured for each outflow tract. When the patient's own atrial contraction occurred during ventricular systole, EjT, AcT, PV and FVI of flow at the LVOT and EjT, AcT and FVI of flow at the RVOT were decreased. Percent change of the FVI of flow at the RVOT (-34.6%) was significantly greater than that of flow at the LVOT (-16.2%, p < 0.01). These results indicate that the loss of right ventricular performance might play a prominent role in the genesis of the hemodynamic deterioration with atrioventricular asynchronous contraction.


[Differentiation between "pseudonormal" from normal transmitral flow velocity waveforms by evaluating isovolumic relaxation time]. We tried to differentiate "pseudonormal" from normal transmitral flow velocity waveforms by evaluating isovolumic relaxation times (IRT) in patients with old myocardial infarction. Forty-three healthy volunteers and 54 patients with old myocardial infarction were studied. Transmitral flow velocity waveforms were obtained by pulsed Doppler echocardiography with a phonocardiogram. Early peak filling velocity (E) and late peak filling velocity (A) were measured, and the E/A ratio was calculated. The time from the beginning of the IIA sound to the onset of transmitral flow was defined as IRT. We observed a significantly positive correlation between IRT and age in the healthy volunteers (r = 0.56, p < 0.01). Based on these results, we selected age-matched healthy subjects (control group, n = 23) older than 35 years from the healthy volunteers. We divided the patients into 2 groups; those with a mean pulmonary capillary wedge pressure (mPCWP) of > or = 16 mmHg (H group, n = 9) and those with an mPCWP of < 16 mmHg (L group, n = 45). E, E/A, IRT, mean blood pressure (mBP), and heart rate were compared among the H, L, and control groups. There was no significant difference in mBP or heart rate between these 3 groups. Both E and E/A were significantly lower in the L group than in the control group (p < 0.05), however, no significant difference was observed in E and E/A between the H and control groups.(ABSTRACT TRUNCATED AT 250 WORDS)


[The effect of heart rate on the time constant of isovolumic relaxation of the left ventricle]. A time constant (T) of the left ventricular isovolumic relaxation, which expresses the relaxation rate of the left ventricle, may be affected by change of the heart rate. There are few clinical reports concerning the relationship between T and heart rate. We studied the relationship between T and heart rate in 10 patients with normal coronary arteries and left ventricular function. Left ventricular pressure was measured using a catheter-tip manometer during right atrial pacing, which was performed at the rate of 10, 20, 40 beats/min in addition to the baseline rate, and finished at 140 or 150 beats/min. Two measurements were made for T.1) Tw from the slope of In (pressure) against time, and 2) Tb by exponential analysis which also estimated the asymptote. As the heart rate increased, both Tw and Tb gradually shortened (Tw: from 38.5 +/- 4.9 msec at the control heart rate to 28.9 +/- 5.6 msec at 140 or 150 beats/min; Tb: from 54.7 +/- 11.4 msec at the control heart rate to 34.9 +/- 5.7 msec at 140 or 150 beats/min). These findings suggest that left ventricular isovolumic relaxation is strongly affected by change of the heart rate in man.


[Conductance catheter for determining left ventricular volume and end-systolic pressure-volume relations: its clinical application]. In this study, the clinical usefulness of the conductance catheter technique was assessed. The end-systolic pressure-volume relations (ESPVR) measured with the increase and decrease in the preload were also compared. Fourteen patients with various heart diseases who underwent diagnostic cardiac catheterization were studied. Using a right atrial pacing catheter, 8 electrical stimuli delivered at a fixed rate of 800 msec were followed by a single early stimulus of 500 msec coupling interval, resulting in premature atrial contraction and post-extrasystolic potentiation (PESP). Left ventricular (LV) volume was measured by 2 methods; namely, the conductance catheter technique and single-plane left ventriculography (LVG). ESPVR was obtained either with the inferior vena cava occlusion induced by balloon inflation, or with the rapid transfusion of saline with electrical resistance identical to the blood volume in the right atrium. The LV volume obtained with the conductance catheter technique (VCON) was correlated with the LV volume as obtained by LVG (VLVG) (end-diastolic volume, VCON = 1.09VLVG - 4.66, r = 0.95; end-systolic volume, VCON = 1.42VLVG - 18.2, r = 0.89). There was correlation of the change in LV volume (delta V) induced by PESP between the 2 methods (end-diastolic volume, delta VCON = 0.52 delta VLVG + 0.94, r = 0.76; end-systolic volume, delta VCON = 0.42 delta VLVG - 4.78, r = 0.72), suggesting that the change in LV volume estimated by the conductance catheter technique was smaller than that estimated by LVG. The LV volume curves were similar.(ABSTRACT TRUNCATED AT 250 WORDS)


[Inverse relationship between coronary flow velocity and myocardial oxygen extraction during intracoronary infusion of papaverine]. The relationship between coronary blood flow velocity (CBFV) and myocardial oxygen extraction (O2-Ext) was investigated during rapid changes of CBFV after intracoronary papaverine infusion. In 6 patients without stenosis of the left anterior descending artery (LAD), one with hypertrophic cardiomyopathy, 2 with syndrome X and 3 with effort angina pectoris, simultaneous measurements of CBFV using the Doppler catheter system and coronary venous oxygen saturation using the fiberoptic catheter system were continuously performed before and during intracoronary infusion of papaverine. When O2-Ext was related to CBFV in every cardiac cycle, there was a good, inverse linear relationship, both in the increase (r = 0.81 +/- 0.24) and decrease (r = 0.93 +/- 0.04) phases of CBFV. The increase in cross-sectional area of segment 6 in the LAD as observed on orthogonal coronary angiograms was 6.0 +/- 2.0%. These results imply that the increase in CBFV during intracoronary papaverine infusion seems parallel to that of coronary blood flow, and that papaverine induces no significant change in myocardial oxygen consumption. Myocardial oxygen extraction in response to changes in coronary flow is regulated readily to meet the myocardial oxygen demand.


[Comparison of treadmill exercise, handgrip, and cold-pressor tests: with particular reference to the effects on hemodynamics, respiratory gas exchange, and sympathetic nervous activity]. The treadmill test (TM), handgrip test (HG) and cold-pressor test (CP) are now frequently used clinically for multiple purposes. However, gas exchange analysis has not been a common procedure during HG. In particular, during CP, it has not been previously reported. Relationships between these 3 tests and blood pressure, heart rate (HR), respiratory gas exchange and the sympathetic nervous activity of normal subjects have not been reported, either. This study was undertaken to clarify these points. Symptom-limited TM was performed in 11 normal male subjects with a mean age of 45 +/- 8 yrs according to the Bruce protocol, with the HG using the weight-sustaining method (equal weight of 50% maximal voluntary contraction) for 3 min, and CP for 2 min. Systolic and diastolic blood pressures (Ps, Pd) were recorded; HR was measured every 30 sec, and gas exchange variables, such as oxygen uptake (VO2) and carbon dioxide production, were documented every 10 sec using an aereomonitor AE-280 (Minato Medical Science Co). In 10 of 11 subjects, concentrations of plasma noradrenaline (PNA) and plasma adrenaline (PAD) were measured at rest and at the times of peak values of the 3 tests. The peak values of Ps and HR were much higher during TM than during HG and CP (p < 0.01), while the peak values of Pd during HG and CP were higher than during TM (p < 0.01). The VO2 increased significantly for all of the 3 tests (TM: +781%, HG: +65%, CP: +20%), with the increment being the greatest during TM. Both PNA and PAD increased significantly for the 3 tests, with the increments of PNA and PAD being the greatest during TM. The percent change in PAD was more prominent during HG and CP than during TM. This tendency was not as clear for PNA as for PAD. There was no correlation of delta Ps and delta Pd between the 3 tests, but values of delta HR correlated partially. No significant correlations of peak VO2 were observed between the 3 tests. The peak PNA correlated between HG and CP (r = 0.77, p < 0.01), and the peak PAD correlated between TM and CP (r = 0.67, p < 0.05). In summary, numerous differences in hemodynamic and respiratory responses and in sympathetic nervous activation were observed in the 3 tests. When the 3 tests are undertaken, careful attention should be paid for their characteristics, discrepancies and limitations.


[Left ventricular wall motion mimicking stunned myocardium in hypertrophic obstructive cardiomyopathy with normal coronary arteries: a case report]. A 63-year-old woman was admitted to the coronary care unit of Hyogo College of Medicine because of cardiogenic shock. She previously had been hospitalized in the Gynecology Department for the treatment of recurrent uterine cancer. She had poor appetite due to chemotherapy which was given for 10 days prior to her admission. On admission, echocardiography and cardiac catheterization revealed hypertrophic obstructive cardiomyopathy and extensive left ventricular wall motion abnormalities. Coronary arteriography showed no coronary artery disease. Left ventriculography as well as echocardiography performed on the 21st post-admission day revealed that the wall motion abnormalities had completely resolved and the systolic anterior motion of the mitral valve (SAM) was no longer evident. The systolic pressure at the apex of the left ventricle was 200 mmHg on admission. The increased ventricular pressure and the simultaneous resolution of the wall motion abnormality and SAM suggest that marked obstruction of the left ventricular outflow tract is more likely to be involved in transient ventricular wall motion abnormality rather than acute myocardial ischemia. The mechanism of the SAM in the present case seemed to be related to a Venturi effect which was augmented by the decreased preload due to hypovolemia. In addition, papillary muscle contraction seemed to pull the mitral valve toward the interventricular septum during systole.


Concentration of plasminogen activators and inhibitor in the human endometrium at different phases of the menstrual cycle. The concentrations of tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) and plasminogen activator inhibitor (PAI-1) have been determined in endometrial curettings obtained from 46 subfertile women during proliferative, early or late secretory phases of the menstrual cycle. t-PA activity and antigen concentrations was significantly higher (P < 0.001) in late secretory endometrium than in proliferative or early secretory endometrium. Higher concentrations of PAI-1 antigen (P < 0.05) were also noted in late secretory phase than in proliferative and early secretory endometrium. However, u-PA concentration was not significantly different and no PAI activity could be demonstrated in the menstrual phases studied. Zymography studies confirmed the presence of both t-PA and u-PA in the endometrium. Ovarian hormonal patterns may therefore influence the activity of plasminogen activators especially of t-PA in the endometrium during various phases of the menstrual cycle.


Synergistic effects of insulin-like growth factor I and gonadotrophins on relaxin and progesterone secretion by ageing corpora lutea of pigs. Insulin-like growth factor I (IGF-I) is involved in paracrine/autocrine regulation of gonadal steroidogenesis and peptide hormone biosynthesis. This study was designed to determine whether IGF-I alone, or an interaction of IGF-I, is involved in augmenting the actions of luteinizing hormone (LH) and prolactin in controlling relaxin and progesterone secretion from ageing corpora lutea of hysterectomized gilts at days 110, 113 and 116 after oestrus. Luteal tissue slices were incubated for 8 h with IGF-I (0, 50, 300 ng ml-1), LH (0, 100, 1000 ng ml-1), and prolactin (0, 100, 1000 ng ml-1) alone or in combination. Progesterone and relaxin concentrations were determined by radioimmunoassay of spent medium and of homogenates from luteal tissue slices before and after incubation. Porcine luteal tissue from day 110 had a net output of 25 ng progesterone and 26 ng relaxin in the control and of 65 ng progesterone and 2125 ng relaxin in the combined IGF-I, LH and prolactin treatment mg-1 of luteal tissue, respectively. IGF-I, LH and prolactin alone or in combination significantly increased (P < 0.01) progesterone production by luteal tissue from day 110, but they were partially effective at day 113 and ineffective at day 116. By contrast, the same hormone treatments increased relaxin production by luteal tissue from days 110 and 113. Even at day 116, prolactin alone or with LH or IGF-I continued to stimulate relaxin production. In conclusion, IGF-I augments the ability of prolactin and LH to increase relaxin production by ageing corpora lutea; however, a decrease in progesterone secretion and an increase in relaxin secretion at day 113 indicate that different mechanisms control progesterone and relaxin secretion in pigs.


Characterization of the motility of maturing rat spermatozoa by computer-aided objective measurement. A computer-aided sperm analysis system was optimized for objective assessment of the movement characteristics of mature and immature rat spermatozoa by testing different settings. Measurements of straight line velocity of individual motile cells were validated by manual tracking with a digitizer. Better agreement between the two methods and better performance in distinguishing between mature and immature spermatozoa was obtained by reducing the tracking rate to increase the time of analysis. However, numbers of motile and immotile cells could not be determined accurately. Manual counting of videotaped images revealed no significant differences in percentage motility of spermatozoa from five epididymal regions. Caput spermatozoa were characterized by low straight-line (VSL) and averaged-path (VAP) velocities and low path straightness (STR), whereas mature cells displayed high VSL, VAP and STR. An increase in curvilinear velocity on maturation was less obvious. Spermatozoa in the proximal corpus epididymidis were heterogeneous in their acquisition of motility maturation and the uniformity of movement pattern achieved in the distal corpus and proximal cauda regions tended to decrease again in the distal cauda epididymidis. Such objective measurements of motility patterns will facilitate studies on the regulation of motility development upon sperm maturation.


Variation in the timing of the reproductive season among breeds of sheep in relation to differences in photoperiodic synchronization of an endogenous rhythm. Photoperiod may regulate seasonal reproduction either by providing the primary driving force for the reproductive transitions or by synchronizing an endogenous reproductive rhythm. This study evaluated whether breed differences in timing of the reproductive seasons of Finnish Landrace (Finn) and Galway ewes are due to differences in photoperiodic drive of the reproductive transitions or to differences in photoperiodic synchronization of the endogenous rhythm of reproductive activity. The importance of decreasing photoperiod after the summer solstice in determining the onset and duration of the breeding season was tested by housing ewes from the summer solstice in either a simulated natural photoperiod or a fixed summer-solstice photoperiod (18 h light:6 h dark; summer-solstice hold). Onset of the breeding season within each breed did not differ between these photoperiodic treatments, but Galway ewes began and ended their breeding season earlier than Finn ewes. The duration of the breeding season was shorter in Galway ewes on summer-solstice hold than on simulated natural photoperiod; duration did not differ between photoperiodic treatments in Finn ewes. The requirement for increasing photoperiod after the winter solstice for initiation of anoestrus was tested by exposing ewes from the winter solstice to either a simulated natural photoperiod or a winter-solstice hold photoperiod (8.5 h light:15.5 h dark). Onset of anoestrus within each breed did not differ between these photoperiodic treatments, but the time of this transition differed between breeds. These observations suggest that genetic differences in timing of the breeding season in Galway and Finn ewes do not reflect differences in the extent to which photoperiod drives the reproductive transitions, because neither breed requires shortening days to enter the breeding season or lengthening days to end it at appropriate times. These findings are consistent with the hypothesis that photoperiod synchronizes an endogenous rhythm of reproductive activity in both breeds and that genetic differences in timing of the breeding season reflect differences in photoperiodic synchronization of this rhythm.


Sex-dependent loss of bisected bovine morulae after culture and freezing. The objective of this study was to determine the post-transfer survival rate of bovine embryos cultured between the time of bisection and freezing. In this experiment 158 morulae were bisected and both portions were cultured for 24-44 h either in vivo after transfer to sheep oviducts (n = 80 morulae) or in vitro (n = 78 morulae) in Ham's F10 medium with 20% fetal calf serum, with bovine oviduct cells or in medium collected from oviduct cultures (conditioned medium). After culture, half of each morula was fixed for cytogenetic sex determination (n = 125) and the other half was frozen. The frozen halves were later thawed and transferred (n = 115) to recipients, who were, if pregnant, slaughtered to determine the sex of the fetus. The culture resulted in better pregnancy rates than those previously reported for embryos frozen immediately after bisection. The sex of 49 (33 males, 16 females) of the fixed demi-morulae was determined, and 38 of the transferred demi-morulae established pregnancies (23 males, 10 females and 5 fetuses that were not recovered). The male:female ratio in in vivo and in vitro culture groups was significantly different from the expected ratio of 1:1 and suggests that manipulation and culture of embryos results in a preferential loss of female embryos.


Circulating gonadotrophins during a period of restricted energy intake in relation to body condition in heifers. Beef heifers, 13 months old, were fed to achieve high (7.6 +/- 0.2 units) or low (3.9 +/- 0.1 units) body condition by feeding them one of two diets for 20 weeks. During week 17 of the growth phase, all heifers were ovariectomized. From week 20 to week 27 (restriction phase), all heifers were fed a daily diet containing 0.071 MJ metabolizable energy kg-1 body weight. At weekly intervals throughout the restriction phase, blood samples were collected at 10-min intervals for 11 h to determine the pattern of secretion of luteinizing hormone (LH), the amount of LH released in response to 750 ng (pituitary responsiveness) and 50 micrograms LH-releasing hormone (LHRH, releasable stores) and mean concentrations of follicle-stimulating hormone (FSH) in the circulation. Body weight declined during the restriction phase in a similar fashion in heifers with high and low body condition and changes in body weight were unrelated to mean concentrations of LH and FSH and frequency of LH pulses. Amplitude of LH pulses and responsiveness to 750 ng LHRH increased in a linear fashion with weight loss in heifers with low but not in those with high body condition. The amount of LH released in response to 50 micrograms LHRH decreased with increasing weight loss in heifers with high but not with low body condition, indicating that releasable pools of LH declined with increased weight loss in heifers with high body condition.(ABSTRACT TRUNCATED AT 250 WORDS)


Pre-prosthetic surgery.

Pre-prosthetic surgery. This paper deals with surgical procedures that can be routinely used in a hospital dental clinic to improve the function of the final prosthesis. This report will discuss alveoplasty, Vestibuloplasty, Fibrous Tuberosity Reduction, Denture Fibrosis, Tori and Exostosis, Labial and Lingual Frenum and Oro-Antral Opening (Table 1).


Putative steroid-binding receptors and nonreceptor components and testicular activity in the lizard Podarcis sicula sicula. Labelled testosterone- and oestradiol-binding molecules have been found in the cytosol and nuclei of lizard testes. DNA-cellulose affinity chromatography was used to separate putative sex-steroid-binding receptors (adhering molecules) and nonreceptor components (nonadhering molecules). A putative androgen receptor (Kd: 10(-10) mol l-1; 3-9 fmol g-1 tissue) was found mainly in the nuclei of testicular cells when actively undergoing spermatogenesis. This suggests that, as in higher vertebrates, testosterone is implicated in spermatogenetic step regulation (meiosis and spermiogenesis) in lizard testis. In the cytosol, testosterone-binding molecules (Kd: 10(-9) mol l-1; 384-784 fmol g-1 tissue) with several properties of androgen-binding proteins are present from autumn to spring. The behaviour of these molecules is consistent with the role assigned to androgen-binding proteins as androgen reservoir. A putative oestrogen receptor is present throughout the sexual cycle, except during the culmination phase (breeding). The putative oestrogen receptor may be involved in the regulation of the first spermatogenetic step (spermatogonia multiplication) and in the induction of post-reproductive refractoriness. This phase is present in temperate-zone lizards. These studies show that the evaluation of sex-steroid-binding molecules is useful in considering the relationships between sex hormones and spermatogenetic activity in the testes of lizards.


Effects of dietary zinc deficiency on gonadotrophin secretion and testicular growth in young male sheep. The hypothesis that the secretion of gonadotrophins would be reduced by zinc deficiency was tested in five groups of four young Merino rams (initial liveweight 22 kg). Four groups were fed ad libitum with diets containing 4, 10, 17 or 27 micrograms Zn g-1. The effects of loss of appetite on the deficient diet was controlled by feeding a fifth group (pair-fed control) at a rate of 27 micrograms Zn g-1, but the amount of feed offered was restricted to that eaten voluntarily by the deficient (4 micrograms Zn g-1) group. Blood was sampled every 20 min for 32 h on two occasions before the treatments were imposed and 96 days later, at the end of the experiment. The rams were injected with gonadotrophin-releasing hormone (GnRH; 10 ng kg-1 i.v.) after each serial sampling, and with naloxone (1 mg kg-1 i.v.) 24 h after the end of the final GnRH test. In the group that were fed the diet with the lowest zinc content, the concentration of zinc in blood plasma was reduced to 18% of that in the pair-fed controls (P < 0.05) and was within the deficient range. The appetite of the deficient rams was half that of the controls fed 27 micrograms Zn g-1 ad libitum and there was no increase in liveweight or testicular diameter during pubertal development. Similar, but smaller, effects were observed in the pair-fed controls. There were no significant differences between pair-fed and deficient groups in the frequency of the luteinizing hormone (LH) pulses or in the concentration of follicle-stimulating hormone (FSH), but the secretion of gonadotrophins was markedly lower in both groups than in the control rams fed ad libitum. The response to GnRH was not affected by treatment, but the increase in LH pulse frequency evoked by naloxone was lower in the deficient animals than in other groups. The animals fed zinc at intermediate rates (10-17 micrograms g-1) showed similar responses to the controls fed ad libitum. It is concluded that the specific effects of zinc deficiency on testicular function were small. Most of the reduction in testicular growth in rams fed a deficient diet was not specifically related to the trace element, but was due to the fall in energy and protein intake caused by the loss of appetite. This leads to a reduction in the frequency of GnRH pulses secreted by the hypothalamus, and to low rates of gonadotrophin secretion by the pituitary gland.


Evidence for a gonadal nonsteroidal factor that specifically inhibits release of luteinizing hormone in ewes. Bilaterally ovariectomized ewes were used to investigate the effect of systemic administration (i.v.) of charcoal-treated aqueous luteal extracts from ovine corpora lutea on plasma concentrations of pituitary gonadotrophins. Jugular blood samples were taken every 15 min at least 5 h before (control period) and 5 h after (treatment period) injection. In Expt 1, the administration of luteal extract from corpora lutea of days 70-76 of pregnancy, but not of the extract prepared from muscular tissue, resulted in a significant decrease of mean concentrations of luteinizing hormone (LH) (P < 0.02) and frequency of LH pulses (P < 0.01). Plasma follicle-stimulating hormone (FSH) concentrations were not affected by injections of either extract. These findings provide the first demonstration of the presence of a nonsteroidal factor in the corpus luteum of midpregnancy that selectively suppresses the secretion of LH. In Expt 2, mean concentrations of LH and FSH and frequency of LH pulses were unaffected by injections of luteal extracts from ovine corpora lutea of days 10-12 of the oestrous cycle or day 15 of pregnancy. These data suggest that some factor(s), probably from the fetoplacental endocrine unit, is required to ensure the production of a significant quantity of the luteal LH-inhibiting factor after day 15 of pregnancy. In Expt 3, treatment of luteal extract from corpora lutea of day 70 of pregnancy with proteolytic enzymes destroyed the LH-inhibiting activity, suggesting the proteic nature of the luteal LH-inhibiting factor. In Expt 4, plasma concentrations of LH were not affected by injection of charcoal-treated extract prepared from fetal cotyledonary tissue of days 110-120 of pregnancy suggesting that the LH-inhibiting factor exclusively originates from the corpus luteum during pregnancy. These experiments provide the first direct evidence for the existence of a potent nonsteroidal factor of luteal origin that specifically inhibits pulsatile secretion of LH, without influencing FSH release in female animals. We propose the term LH-release-inhibiting factor (LH-RIF) to describe this activity.


A method to obtain avian germ-line chimaeras using isolated primordial germ cells. Primordial germ cells (PGCs), collected from the blood of 2-day-old chick embryos, were concentrated by Ficoll density centrifugation. The blood contained 0.048% PGCs and the concentrated fraction contained 3.9% PGCs in blood cells. The PGCs were picked up with a fine glass pipette, and one hundred were then injected into the terminal sinuses of 2-day-old Japanese quail embryos (24 somites); bubbles were then inserted to prevent haemorrhage. The embryos were further incubated at 38 degrees C for 24 h, and then fixed. Serial sections were stained with the periodic acid-Schiff reagent (PAS) to demonstrate chicken PGCs and with Feulgen stain to identify quail cells. On the basis of the differences in staining properties, 63.6 +/- 5.3 chick PGCs were detected in the quail embryo in the area where the gonads develop. Furthermore, 39.3 +/- 4.5 chick PGCs were incorporated into the quail germinal epithelium within 24 h of the injection. A similar percentage of the host (quail) PGCs had also migrated to the germinal epithelium at the same stage of development. This technique for obtaining germ-line chimaeras will facilitate research on avian germ-line differentiation.


Cell-mediated immune response after vasectomy in rats. This study investigated the development of a cell-mediated immune response after vasectomy in Swiss Albino rats, by comparing the development of the thymus-dependent lymphoid tissue of the regional testicular lymph node and the spleen in vasectomized and in sham-operated control animals. Frozen sections were used and thymus-dependent regions were stained by immunocytochemistry. After vasectomy, the areas occupied by the paracortex in the lymph node sections showed a significant increase in size; the thymus-dependent regions of the spleen, in contrast, showed no change. The regional lymph node, rather than the spleen, seems to be important in cell-mediated and humoral immune responses to vasectomy.


Heritable testicular hypoplasia in Nguni (Bos indicus) bulls: vascular characteristics and testosterone production. The biased unilateral occurrence of heritable gonadal hypoplasia was investigated by examining the gross- and microanatomy of the testicular artery and vein, testicular blood flow and testicular testosterone secretion in normal Nguni bulls and in Nguni bulls showing unilateral left, unilateral right and bilateral hypoplasia of the testis. A high incidence of branching of the testicular artery was found ipsilateral to hypoplastic testes. The branching occurs a short distance from the dorsal aorta: one branch proceeds to the testis, the other to the ipsilateral kidney. The association between arterial branching to the kidney and ipsilateral hypoplasia of the testis held for both unilaterally left and unilaterally right hypoplastic bulls. Variations in the anatomy of the testicular vein occurred in both normal and hypoplastic bulls but there was no specific association between the variations and ipsilateral hypoplasia. The lumen diameter of the testicular artery or branch correlated with testis mass. Wall thickness of the artery ipsilateral to hypoplastic testes was not different from that in normal bulls, discounting hyperplasia of the endothelium. Total blood flow to the testis correlated with testis mass. The secretion rate of testosterone from hypoplastic testes was lower than that of normal testes but there was no difference when compared on a unit mass basis.


Effect of surgical restriction of growth of the testicular artery on testis size and histology in bulls. The growth of the testicular artery was restricted on one side in young bulls and subsequent testicular development was monitored. After the animals had been killed, the testes were studied histologically and compared with testes from hypoplastic bulls. The growth rate of testes from the experimental side was significantly lower than that of testes from the sham-operated side over a period of 578 days. At death, the experimental testes had a mean (+/- SD) mass of 76 (+/- 41) g compared with 220 (+/- 31) g for the control testes. The sham-operated testes accounted for 0.071 (+/- 0.008)% of live body mass compared with 0.025 (+/- 0.014)% for the experimental testes. The seminiferous tubules in the sham-operated testes had a mean diameter (+/- SD) of 211 (+/- 25) microns, whereas those of the artery-restricted testes and hypoplastic testes were significantly smaller (152 (+/- 37) and 145 (+/- 45) microns, respectively). In the artery-restricted and hypoplastic testes, the interstitial tissue accounted for a significantly greater proportion of the testicular tissue than in the sham-operated testes and spermatogenesis was either totally absent or present in only a small proportion of tubules. It is suggested that the artery-restricted testes could be used as a model for testicular hypoplasia.


Ability of thawed tiger (Panthera tigris) spermatozoa to fertilize conspecific eggs and bind and penetrate domestic cat eggs in vitro. Electroejaculates from tigers were collected and half was used fresh to inseminate tiger eggs in vitro and domestic cat eggs stored in a hypertonic salt solution. The remainder was pelleted, frozen in a solution of 20% egg yolk, 11% lactose and 4% glycerol, thawed and cultured with tiger and domestic cat eggs. The motility index ((sperm % motility)+(status rating x 20))/2 for thawed spermatozoa was about 86% of that in fresh aliquots. Of the 49 tiger oocytes inseminated in vitro with fresh spermatozoa, 34 (69.4%) cleaved, compared with 33 of 47 oocytes (70.2%) cultured with thawed spermatozoa (P > 0.05). Embryos generated by either sperm treatment could develop in vitro to the 16-cell or morula stage. Fresh and thawed tiger spermatozoa were equally capable (P > 0.05) of binding and penetrating the outer and inner zona pellucida of domestic cat eggs. These results demonstrate the ability of frozen-thawed tiger spermatozoa to (i) penetrate homologous and heterologous eggs and (ii) result in conspecific, advanced development of preimplantation embryos in vitro.


Ability of in vitro maturing bovine oocytes to transform sperm nuclei to metaphase chromosomes. Bovine oocytes at the germinal vesicle stage were inseminated in Brackett & Oliphant's medium with bovine serum albumin, caffeine and heparin. Eight hours after insemination, oocytes were transferred into tissue culture medium-199 containing 10% fetal calf serum and cultured for 5-40 h at 39 degrees C in 5% CO2 in air. The proportions of unpenetrated and penetrated oocytes reaching metaphase II increased as the time of examination increased, reaching 70 and 65% 40 h after transfer, respectively. When oocytes were penetrated by more than four spermatozoa, meiotic maturation was greatly retarded. Sperm nuclei were decondensed in most (81%) penetrated oocytes 5 h after transfer. The decondensed sperm nuclei were recondensed and then transformed to metaphase chromosomes which were morphologically compacted at first but became slightly dispersed later. The formation of the metaphase chromosomes was observed in 86% of penetrated oocytes examined 40 h after transfer, and occurred in all metaphase II oocytes at that time. In oocytes penetrated by more than nine spermatozoa, no such transformation of sperm nuclei was observed. Well-developed male and female pro-nuclei were observed in only three (6%) of 51 oocytes penetrated 40 h after transfer.


Expression of activities of two 20 alpha-hydroxysteroid dehydrogenase isozymes in rat corpora lutea. The rat ovary contains two isozymes of 20 alpha-hydroxysteroid dehydrogenase (HSD-1 and HSD-2). In this study, the expression of activity of each isozyme was investigated in ovaries that contained a single generation of corpora lutea during pseudopregnancy. This condition was induced by cervical stimulation in rats that had been rendered anovulatory by housing them in a continuously lit environment. The total activity of cytosolic 20 alpha-HSD was lower in the ovaries of these pseudopregnant rats than in ovaries containing multiple generations of corpora lutea. In normal pseudopregnancy, HSD-1 activity was low on days 5 and 9 and increased markedly on day 15, whereas HSD-2 was lower than HSD-1 and did not vary throughout pseudopregnancy. However, on days 5 and 9 of continuous-light pseudopregnancy, low activity of HSD-1 only was detected; by day 15, HSD-1 activity had increased sixfold and HSD-2 activity could be detected. Immunohistochemical methods using a specific antibody recognizing both HSD-1 and HSD-2 revealed that the number of 20 alpha-HSD-positive luteal cells increased by day 15. Thus, the increase in total enzyme activity and appearance of HSD-2 activity observed at late pseudopregnancy was accompanied by an increase in the number of 20 alpha-HSD-positive luteal cells.


Binding of a 15 kDa glycoprotein from spermatozoa of boars to surface of zona pellucida and cumulus oophorus cells. A highly purified 15 kDa glycoprotein isolated from ejaculated spermatozoa was used to raise antisera in female rabbits. An indirect immunofluorescence technique was used to detect the antigen in the seminal vesicle tissue and on the acrosomes of ejaculated, native and capacitated, boar spermatozoa. No immunoreactivity was detected on cells of the seminiferous tubules (spermatogonia, spermatocytes, and spermatids), on spermatozoa in the ductus epididymis and in cells of the epididymal and testicular tissues. These observations support the view that the 15 kDa protein is produced in the seminal vesicle secretory epithelium, and is attached to the sperm plasma membrane during the exposure of spermatozoa to seminal vesicle compounds. The observations that the antigen remained on the acrosome of ejaculated spermatozoa after capacitation and blocked sperm-oocyte binding in vitro suggest that the antigen plays a role in sperm-egg interactions. The strong immunoreactivity exhibited by cumulus cells after incubation of antisera with the porcine egg surrounded by cumulus cells shows the possible importance of the 15 kDa glycoprotein for contact of spermatozoa with cells of the cumulus oophorus surrounding the egg.


Evidence for the action of bovine follicular fluid factor(s) other than inhibin in suppressing follicular development and delaying oestrus in heifers. The aim of this study was to investigate the importance of inhibin in the delay in return to oestrus in heifers induced by steroid-stripped bovine follicular fluid (bFF). Oestrous activity was synchronized in 18 Hereford x Friesian heifers with two injections of prostaglandin (PG) 12 days apart. At the time of the second PG injection (time 0), the animals were assigned at random to one of three experimental groups and received i.v. injections of 20 ml saline (controls, n = 6), whole bFF (FF group, n = 6) or bFF in which the bioactive inhibin content had been reduced by > 95% by immunoaffinity chromatography (-INH group, n = 6; inhibin content approximately 0.8 ml whole bFF) every 8 h for 2 days. In a dose-response study, 2.5 ml whole bFF was insufficient to delay oestrus consistently following a similar synchronization regimen. Blood samples were taken every 8 h, initially before each injection and then subsequently for a further 9 days for hormone analysis. Animals were observed every 8 h throughout the experiment for signs of behavioural oestrus. The ovaries of all animals were examined using real-time ultrasonography about 30 h after the second PG injection. Treatment failed to suppress peripheral follicle-stimulating hormone (FSH) concentrations, although a significant increase was observed in both treatment groups after cessation of injections. Progesterone concentrations fell immediately after the second PG injection in all animals and remained below minimum detectable concentrations in all treated animals for the remainder of the experiment. In control animals, progesterone rose above minimum detectable concentrations by day 6 and continued to rise until the end of the experiment. Analysis of samples taken from treated animals several days after observed oestrus revealed that all had apparently ovulated. Mean daily luteinizing hormone (LH) concentrations did not differ between treatment groups before ovulation, but after ovulation, mean daily LH was significantly reduced in control animals as progesterone concentrations rose. Follicular development, as assessed by the mean antral diameter of the largest follicle on a pair of ovaries at ultrasound examination, was significantly suppressed in treated animals compared with controls (P < 0.01) and there was no significant difference (P = 0.397) between the two treatment groups. Control animals displayed oestrus 68 h (+/- 8 SEM) after the second PG injection, but oestrus was delayed in treated animals to 186h +/- 5 (FF group) and 191 h +/- 6 (-INH group).


Uptake and incorporation of inositol by preimplantation mouse embryos. The uptake of myo-inositol by preimplantation mouse embryos was investigated using [3H]myo-inositol. Uptake increased about 12-fold between one- and two-cell stages and increased again at the blastocyst stage (> 6-fold compared with the two-cell stage). Uptake at the blastocyst stage was time and temperature dependent; it was stimulated by sodium, inhibited by glucose and appeared to take place mainly via a saturable mechanism. Uptake in the presence of 6.25 mmol inositol l-1 was 1424 fmol inositol per blastocyst per h. About 10% of the [3H]inositol taken up by blastocysts during 8 h in culture was incorporated into lipid. Thin layer chromatography of the lipid showed that most of this inositol was incorporated into lipid material co-migrating with phosphatidylinositol with a small proportion co-migrating with phosphatidylinositol 4-phosphate.


Effect of progesterone on ovarian follicles, emergence of follicular waves and circulating follicle-stimulating hormone in heifers. The hypothesis was tested that greater growth of the dominant follicle of wave 1 (first follicular wave of an interovulatory interval), compared with that of subsequent anovulatory waves, is due to lower circulating concentrations of progesterone during the growing phase of the follicle. Control heifers (n = 6) were compared with heifers (n = 6) treated with a decreasing dose of progesterone from day 0 to day 5 (ovulation = day 0). Maximum diameter (12.7 +/- 0.9 versus 15.3 +/- 0.7 mm) and mean diameter of the dominant follicle of wave 1, averaged over days, were smaller (P < 0.05) in the progesterone-treated than in the control group. Progesterone treatment did not suppress circulating follicle-stimulating hormone (FSH); but the second FSH surge was earlier, resulting in earlier emergence of wave 2 as indicated by a tendency (P < or = 0.1) for group x day interactions attributed to earlier detection of the dominant follicle and an earlier rise in the total number of follicles detected. The stated hypothesis was supported. We also tested the hypothesis that exposure to low circulating concentrations of progesterone at the end of the growing phase of the anovulatory dominant follicle of wave 1 results in continued growth and prolonged maintenance of the dominant follicle. Heifers (n = 6 per group) were given a luteolytic dose of prostaglandin F2 alpha (PGF2 alpha) on day 6 and treated with a low (30 mg day-1), physiological (150 mg day-1), or high (300 mg day-1) dose of progesterone on days 6 to 20. Continued periodic emergence of anovulatory follicular waves occurred (2.1 +/- 0.0 waves, 2.8 +/- 0.2 waves, 3.8 +/- 0.3 waves, respectively; P < 0.05) until treatment was stopped (interovulatory intervals: 26.2 +/- 1.0, 30.8 +/- 0.6 and 40.3 +/- 1.7 days, respectively; P < 0.05). Compared with the physiological dose group, the growth of the dominant follicle was inhibited to a lesser degree in the low-dose group since it grew for longer (P < 0.05) and to a larger diameter (P < 0.05), and persisted for longer (P < 0.05). Prolonged dominance of this oversized (> 20 mm) follicle was associated with delayed emergence of wave 2. The hypothesis was supported. Results also showed that the high dose of progesterone suppressed the dominant follicle more than the physiological dose when given during the growing phase, but not when given after the growing phase.(ABSTRACT TRUNCATED AT 400 WORDS)


Insulin and insulin-like growth factor I in follicular fluid after induction of ovulation in women undergoing in vitro fertilization. This study was undertaken to evaluate the relationship between concentrations of insulin and insulin-like growth factor I (IGF-I) in follicular fluid and fertilization and cleavage of human oocytes fertilized in vitro. The concentration of oestradiol, progesterone, luteinizing hormone, follicle-stimulating hormone, testosterone, insulin and IGF-I was determined in 36 follicular fluids, free of visible blood contamination and containing mature oocyte-corona-cumulus complexes, obtained from 12 women undergoing in vitro fertilization. Follicular development was induced by clomiphene citrate and human menopausal gonadotrophin, and follicular aspiration was performed 35 h after an ovulatory dose of human chorionic gonadotrophin. Concentrations of IGF-I were significantly higher in follicular fluids associated with mature oocytes that fertilized and cleaved, than in follicular fluid associated with mature oocytes that did not fertilize (P < 0.001). There was no difference in the concentration of insulin between follicular fluids from which fertilized oocytes were obtained and those with oocytes that remained unfertilized. No significant correlations were found between rates of embryo cleavage, concentrations of insulin and IGF-I. Multiple linear regression analysis demonstrated that the concentrations of IGF-I in follicular fluid were predicted statistically by a negative regression coefficient for the concentration of testosterone, and by a positive regression coefficient for the concentration of progesterone in follicular fluid. No candidate variable was included in the model to predict concentrations of insulin. These data suggest an important role for IGF-I in the mature follicle.


Adverse effects of gonadotrophin treatment on pre- and postimplantation development in mice. The effect of gonadotrophins on pre- and postimplantation development in mice was investigated by superovulating C57BL/6J/Bom females with pregnant mares' serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG) or by inducing ovulation with hCG. In both hormone treated groups, the proportion of abnormal preimplantation embryos increased compared with naturally ovulating animals. Postimplantation mortality increased and the mean number of live fetuses per pregnant mouse decreased in superovulated and hCG-treated mice compared with controls. Embryonic growth was highly retarded. Mean weight of live fetuses in superovulated and hCG-treated mice was reduced and skeletal examination revealed developmental retardation. In conclusion, superovulation as well as induction of ovulation adversely affected embryonic and fetal development.


Effect of oxytocin infusion on secretion of progesterone and luteinizing hormone and the concentration of uterine oxytocin receptors during the periovulatory period in cloprostenol-treated ewes. Oxytocin infusions were initiated on day 10 of the oestrous cycle in ewes, and luteal regression was induced by injection of 100 micrograms cloprostenol on day 12. Blood samples were collected at frequent intervals via an indwelling jugular vein cannula to measure concentrations of progesterone and luteinizing hormone (LH) during the luteal and follicular phases in saline (n = 6) and oxytocin (n = 5) infused animals. The oxytocin infusion maintained peripheral plasma concentrations of 53 +/- 3.2 pg oxytocin ml-1 (mean +/- SEM) compared with values of about 1 pg ml-1 during oestrus in control ewes. Oxytocin infusion had no effect on luteal phase progesterone concentrations, the timing of luteolysis, basal luteinizing hormone (LH) secretion, LH pulse frequency, or the timing or height of the LH surge. Treated ewes came into oestrus significantly earlier than controls (P < 0.05) but ovulated normally. Uterine samples collected 96 h after cloprostenol injection (approximately day 2 of the cycle) showed that oxytocin receptor concentrations were significantly higher in the endometrium in ewes that had been given a 5 day oxytocin infusion than in control animals (556 and 262 fmol mg-1 protein, respectively: geometric means from ANOVA, P < 0.001), whereas myometrial receptor concentrations were not affected (113 and 162 fmol mg-1 protein, respectively). We conclude that the previously reported delay in luteal development caused by oxytocin infusion (Wathes et al., 1991) is not due to the inhibition or delay of ovulation, but must instead occur via a direct influence on the developing corpus luteum.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of age and time of day on the timing of the surge in luteinizing hormone, behavioural oestrus and mating in red deer hinds (Cervus elaphus). The oestrous cycles of fourteen red deer hinds (six yearling; eight more than 2 years old) were synchronized during the early breeding season by removal of a progesterone-containing intravaginal device and blood samples were taken at intervals of 3 h commencing 13 or 25 h later and continued for 54 h. The controlled internal drug release devices (CIDRs) were removed at 08:00 h (group 1; three yearlings and four adults) or 12 h later at 20:00 h (group 2; three yearlings and four adults). There was no significant effect of time of removal of CIDR on the interval to the onset of oestrus (group 1, 34.5 +/- 4.05 h; group 2, 42.14 +/- 7.8 h) on the time of peak concentration (group 1, 41.81 +/- 5.69 h; group 2, 41.71 +/- 7.81 h) or on duration of the luteinizing hormone (LH) surge (group 1, 15.00 +/- 0.95 h; group 2, 14.57 +/- 0.78 h). The six yearling animals exhibited oestrus and LH surge significantly later than the adults (55 +/- 4.2 versus 32 +/- 6.3 h for the LH surge for yearling and adult females, respectively). In a further experiment, 20 hinds were synchronized during the breeding season by removal of CIDR at two times of day 12 h apart and placed with a stag. Mating took place at a mean time of 42.1 +/- 2.4 h and 37.0 +/- 1.3 h later in the two groups. There was no significant effect of time of removal of CIDR upon time to onset of oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)


In vivo microscopy of the subepithelial capillary plexus of the endometrium of rats during embryo implantation. Using in vivo microscopy we investigated endometrial microvascular events occurring on days 5 and 6 of pregnancy at the time of implantation. Blood flow through the endometrium was visualized using incident-light fluorescence microscopy and a video image was recorded for subsequent analysis. At 17:00 h on day 5 of pregnancy it was not possible to identify the impending implantation site from the in vivo appearance of the subepithelial capillary plexus. At 09:00 h on day 6 of pregnancy the embryo implantation site was recognized as an avascular area surrounded by large diameter vessels. These were highly susceptible to haemorrhage when handled. Capillaries closest to the embryo had the greatest diameters, averaging 18.5 +/- 2.5 microns, and capillary diameters decreased to 7.5 +/- 0.4 microns by 2000 microns from the embryo. It was also observed that blood flow through larger diameter vessels was sluggish with frequent reversals and stoppages. Leucocyte rolling and adhesion were also common features in these larger vessels. These data indicate that changes in capillary diameter occur in response to local signals associated with the implanting rat embryo. The embryonic or local endometrial signals that mediate these major microvascular changes remain to be elucidated.


Requirement for glucose in ligand-stimulated meiotic maturation of cumulus cell-enclosed mouse oocytes. In this study, the effect of different energy sources used in Eagle's minimum essential medium on the meiotic maturation of mouse oocytes in culture was examined. The effects of glucose (5.5 mmol 1(-1)), pyruvate (0.23 mmol 1(-1)) and glutamine (2 mmol 1(-1)) in different combinations were tested on the maturation of denuded oocytes in the presence or absence of 300 mumol dibutyryl cAMP 1(-1) during 17-18 h of culture. In the absence of cyclic nucleotide, only oocytes from those groups containing pyruvate resumed maturation at a high frequency (99-100% germinal vesicle breakdown); all other combinations resulted in < or = 54% germinal vesicle breakdown. When dibutyryl cAMP was introduced, all pyruvate-containing groups exhibited maturation frequencies of about 50%, whereas maturation in all other groups was negligible (< or = 10% GVB). Pyruvate was also important for the maintenance of viability in denuded oocytes (> or = 86% viability in pyruvate-containing medium; < or = 35% viability in pyruvate-free groups). When cumulus cell-enclosed oocytes were cultured in medium without inhibitor, all combinations of energy substrates supported high frequencies of maturation (> or = 89% germinal vesicle breakdown) and viability (> or = 91%). The addition of dibutyryl cAMP resulted in inhibition of meiotic maturation (5-33% germinal vesicle breakdown) in all cultures except the pyruvate-alone group (97% germinal vesicle breakdown). Viability in cumulus cell-enclosed oocytes was greatest when two or more energy substrates were present in the medium. Follicle-stimulating hormone (FSH) produced a stimulation of meiotic maturation in all cultures of meiotically arrested cumulus cell-enclosed oocytes, but maximal induction of germinal vesicle breakdown was dependent upon D-glucose. Concanavalin A (ConA)-induced meiotic maturation was also dependent upon D-glucose. Uptake and metabolism of D-glucose by the cumulus cells is important in mediating the stimulatory effects of these ligands on oocyte maturation because (1) both FSH and ConA stimulated uptake of D-glucose and 2-deoxyglucose but not 3-O-methylglucose; (2) phloretin prevented the stimulatory action of FSH and ConA on germinal vesicle breakdown at a concentration that suppressed ligand-induced uptake of D-glucose; (3) 2-deoxyglucose, a hexose that suppresses glycolysis, prevented the induction of meiotic maturation by FSH and ConA and (4) D-mannose, a glycolysable sugar, was as effective as D-glucose in supporting the ligand effects.(ABSTRACT TRUNCATED AT 400 WORDS)


Concentrations of energy substrates in oviductal fluid and blood plasma of pigs during the peri-ovulatory period. Large White gilts, 9 to 18 months old, that had exhibited at least two natural oestrous cycles were divided into three groups (phases): unmated pre-ovulatory, unmated post-ovulatory and mated post-ovulatory (n = 16, 20 and 18). Oviductal luminal fluid samples were collected under anaesthesia by micropipette from the ampulla and ampullary-isthmic junction and analysed by an ultramicrofluorometric technique. Glucose concentrations (mmol 1(-1), means combining regions; mean +/- SEM) were significantly higher in blood plasma than in oviductal fluid (4.56 +/- 0.20 versus 0.59 +/- 0.16; P < 0.0001; n = 27), whereas lactate was higher in the oviduct (5.71 +/- 0.53 versus 2.48 +/- 0.24; P < 0.0001; n = 27). No significant differences were found between the ampulla and the ampullary-isthmic junction. However, the concentration of glucose was significantly higher (P < 0.05) in the ampulla of the pre-ovulatory group (0.97 +/- 0.20; n = 13) compared with the mated group (0.25 +/- 0.05; n = 14) and its concentration in the ampullary-isthmic junction in the pre-ovulatory group (1.65 +/- 0.63; n = 13) was significantly greater (P < 0.05) than in the post-ovulatory (0.43 +/- 0.11; n = 11) or mated groups (0.17 +/- 0.02; n = 14). Lactate in the ampulla of mated animals was higher than in the pre-ovulatory group (6.83 +/- 0.70 versus 3.86 +/- 0.38; P < 0.05; n = 15 and 13), but neither was significantly different from the post-ovulatory group. Furthermore, no change was seen at the ampullary-isthmic junction in lactate concentration with phase.(ABSTRACT TRUNCATED AT 250 WORDS)


Transport of steroids between fetuses via amniotic fluid in relation to the intrauterine position phenomenon in rats. In litter-bearing mammals, the course of development of male and female fetuses is affected by the presence of other fetuses of the same or opposite sex located nearby within the uterus. The transport of testosterone between rat fetuses was examined by implanting a Silastic capsule containing [3H]testosterone into the amniotic sac of a fetus at either the ovarian or cervical end of a uterine horn on days 19 and 20 of pregnancy. The amount of testosterone that was recovered from the amniotic fluid of other fetuses 12 h later was determined. The amniotic fluid surrounding the adjacent fetus on the cervical side of the implanted fetus contained three times as much [3H]testosterone as did the adjacent fetus on the ovarian side, regardless of where in the uterus the implant was made. The movement of dye injected into the uterine lumen was towards the cervix. Intraluminal fluid movement may thus mediate the greater transport of [3H]testosterone towards the cervix than towards the ovary. Our findings support the hypothesis that transport of testosterone between fetuses occurs across the fetal membranes via diffusion, such that any fetus (male or female) located between male fetuses receives the greatest supplement of testosterone.


Collection of oocytes and production of blastocysts in vitro from individual, slaughtered cows. On four occasions ovaries from a total of 35 cows were collected separately at the abattoir where they had been killed. The age of 20 of these cows was recorded. Oocytes from these ovaries were collected separately and were submitted to in vitro maturation, in vitro fertilization and in vitro culture procedures. Ovaries of 34 randomly chosen cows were pooled and treated as the control. Ova from individual cows were cultured in 10 microliters droplets and those from pooled ovaries were cultured in groups of 50 in 50 microliters droplets of oviductal cell-conditioned medium. The 35 cows treated individually supplied 493 oocytes (mean 14.1 oocytes per cow) with high individual variation (SD = 10.0; range = 0-38) and 47 expanded blastocysts (9.5% of oocytes; mean 1.3 blastocysts per cow; range = 0-6). Among these cows, 16 produced one or more blastocysts. Considerable variation in average development rates was detected over the four replicate experiments (11.3, 4.0, 9.0 and 13.5%). The 34 cows treated as the control supplied 397 oocytes (mean 11.7 oocytes per cow) and 44 expanded blastocysts (11.1% of oocytes; mean 1.3 blastocysts per cow) with high variations between replicates (11.1, 4.0 and 18.1%). No difference was observed between individual and pooled ovaries regarding either the number of oocytes, the rate of blastocyst formation, or the number of blastocysts per cow. No effect of age was detected.(ABSTRACT TRUNCATED AT 250 WORDS)


Development of pronuclei in pig oocytes activated by a single electric pulse. Pig oocytes were matured in vitro in a modified M-199 medium for 44 h, subjected to electrical stimulation and scored for activation 6 h later. Sham pulsed oocytes, exposed to electroporation medium and an a.c. field, did not develop the female pronucleus any more frequently than occurs spontaneously (8.3% within 50 h of culture). However, a single d.c. pulse proved extremely efficient in activating pig oocytes. Pulses of 0.75-1.65 kV cm-1 lasting 30 or 100 microseconds activated at least 90% of matured oocytes. The developmental pathway taken by the activated oocytes depended on the parameters of the pulse. The lowest effective stimulation (0.45 and 0.60 kV cm-1 for 30 microseconds) frequently produced oocytes that remained in pre-pronuclear stages of activation (29.4 and 42.3%, respectively). Extrusion of the second polar body and creation of one pronucleus was the most frequent type of activation (in up to 88.2% among the activated oocytes). The strongest stimulations used (1.05-1.65 kV cm-1 for 100 microseconds) often yielded oocytes that failed to extrude the second polar body and formed two or more pronuclei (up to 56.3%). Under optimal stimulation (0.75 kV cm-1), the activated oocytes proceed synchronously to interphase of the first mitotic division. Anaphase II is reached within 30 min and telophase Ii at 1 h after application of the pulse. The second polar body is extruded about 2 h after activation. Well-defined swelling pronuclei were found in oocytes 5-6 h after activation. The relationship between the stage of oocyte maturation and susceptibility to activation was investigated. The period of culture in which the oocytes develop the activation competence (32-36 h of culture) overlapped with the period in which the oocytes complete meiosis (28-38 h). This suggests that ageing in meiotic arrest is not essential for pig oocytes to become activated by electric pulses. Activation of pig oocytes was accompanied by release of cortical granules. In sections of control (metaphase II) oocytes, an average of 7.3 intact cortical granules per 10 microns of overlying cytoplasmic membrane was found. This number dropped to 1.5 in 10 microns within 30 min after the pulse.


Effects of low molecular weight oviductal factors on the development of mouse one-cell embryos in vitro. The relationship between the oviduct and embryo development in the mouse was investigated and the period at which the influence of oviduct can be concerned in the development of mouse embryos in vitro was identified. In addition, the relative molecular weight of oviductal factors that promote embryo development was demonstrated. Mouse zygotes developed to the blastocyst stage when co-cultured with ampulla. The period of embryo co-culture significantly affected the further development of the embryos. Fewer one-cell embryos co-cultured with dissected ampullae for less than 24 h developed to blastocysts than those co-cultured for more than 28 h (P < 0.001). A high percentage of embryos co-cultured with ampullae after 24 h of culture in vitro developed to the blastocyst stage, which suggests that the influences of ampulla on the development of mouse embryos are restricted to a specific period at the two-cell stage (about 55-56 h after hCG injection) in vitro. Mouse ova that were cultured in media conditioned by ampullae could also develop to the blastocyst stage. The fractionated medium that contained low molecular weight fractions was more effective (P < 0.001) on the development of embryos to the blastocyst stage than that containing high molecular weight fractions. These results suggest that the low molecular weight oviductal factors play an important role in the development of mouse embryos at a certain critical age in vitro.


Influence of season of birth on onset of gonadotrophic and ovarian functions in young doe hares (Lepus europaeus). The pituitary and ovarian responses to a monthly i.v. injection of 5 micrograms luteinizing-hormone-releasing hormone (LHRH) were studied in three groups of young doe hares, born in January-February (group I), in April (group II) or at the end of the breeding season (August-September, group III). The LHRH injection was always followed by a release of LH and progesterone, which did not differ among the three groups at 3 months of age. The pituitary and ovarian responses to LHRH increased gradually from the age of 3 months in groups I and III and from the age of 9 months in group II. One female of the ten born in January-February ovulated and reached puberty in June, at the age of 4 months, but with a weak pituitary response. The females born in April displayed a seasonally delayed puberty, at 9 months of age (two of five females ovulated in the next January). Four of the five females born at the end of the breeding season ovulated after LHRH when 5 months old (in February), with a full pituitary-ovarian response. The low pituitary response of group I in June-August, even if 10-20% of females ovulated after LHRH, suggests a need for a period of short days. Then, the most favourable conditions for the hare to reach puberty would be a period of short decreasing daylengths during the fall, followed by increasing daylengths after the winter solstice.


Unexpected oocyte growth after follicular antrum formation in four marsupial species. During examination of maturing preovulatory marsupial oocytes we noted that oocyte diameters were invariably about 50% greater than the figures reported in earlier histological studies. As all previous investigations were limited to small follicles (at most 25% the size of the ovulating follicle), the present study was initiated to examine oocyte growth during the whole period of follicular development. Oocyte and follicle diameters were measured for three Australian (Trichosurus vulpecula, Macropus eugenii and Bettongia penicillata--fresh nonfixed material) and one American marsupial species (Monodelphis domestica--histological sections) in which multiple follicle development had been induced by exogenous gonadotrophin treatment. In all species oocytes were obtained from follicles ranging from pre-antral to immediately pre-ovulatory (maximum follicle sizes obtained were: T. vulpecula, 4.5 mm; M. eugenii, 4.3 mm; B. penicillata, 2.5 mm; M. domestica, 0.7 mm). In two of the species (T. vulpecula and B. penicillata) ovulated oocytes were also examined. In T. vulpecula and M. eugenii oocytes were found to achieve much greater diameters than previously reported from histological studies of small follicles (< 0.8 mm) and similar patterns of growth were found in the other two species. In the four species oocytes reached diameters about two to three times that found for eutherian mammals. It was concluded that the marsupial oocyte continued to grow after formation of the follicular antrum and that, although the rate of oocyte growth slowed in larger follicles, it continued into the period immediately before ovulation. In B. penicillata the largest oocytes were obtained after ovulation.(ABSTRACT TRUNCATED AT 250 WORDS)


Seasonal reproduction of a tropical bat, Anoura geoffroyi, in relation to photoperiod. Anoura geoffroyi (Chiroptera, Phyllostomidae, Glossophaginae), Geoffroy's hairy-legged long-tongued bat, were collected from September 1984 to August 1985, and these bats were found to breed seasonally in the wild on Trinidad, West Indies, at 10 degrees N latitude. Histological examination of these samples indicated that females became pregnant in July or August, and young were born in late November or early December. The testes and epididymides were small from September to mid-April, increased threefold in weight between mid-April and late May, reached a peak weight in July, and decreased in weight in August. Spermatogenesis occurred throughout the testes of males captured from May to August. In 1990, the timing of parturition in females that gave birth in the laboratory to young conceived in the wild was similar to the timing in the field in 1984-1985. Groups of 10-13 males were subjected in the laboratory to (i) a gradually changing, civil twilight photoperiod that mimicked the natural cycle of annual change at 10 degrees N latitude, (ii) the same gradually changing cycle of photoperiod accelerated to a six-month period, or (iii) a constant photoperiod (light 12:54 h: dark 11:06 h). These treatments began in mid-December, four months before the initiation of testicular recrudescence in the wild. In all three groups, testicular volume remained low until April, and then increased two- to threefold between late April and late June, rising to a peak in July, as occurred in the wild.(ABSTRACT TRUNCATED AT 250 WORDS)


Concentration of oxytocin receptors in the placenta and fetal membranes of cows during pregnancy and labour. The concentrations of oxytocin receptors were measured in intercaruncular and caruncular endometrium, fetal cotyledons, chorioallantois and amnion during pregnancy and parturition in cows. Tissues were obtained on days 20 (endometrium only), 50, 100, 150, 200, 225, 250, 275, at term (days 280-284), during labour and within 24 h after calving. Receptor concentrations in intercaruncular endometrium were low on day 20 of pregnancy, 39 +/- 11 fmol mg-1 protein. By day 50, receptor concentrations had increased more than tenfold to 572 +/- 52 fmol and rose steadily until day 250 and then levelled off at about 4500 fmol mg-1. Shortly before parturition, on day 282 +/- 1, a further rise to 7300 +/- 1418 fmol mg-1 was observed, these concentrations were maintained throughout labour. By contrast, caruncular endometrial receptor concentrations remained low until term, mean 145 +/- 15 fmol mg-1, and then rose to 720 +/- 163 fmol mg-1 during labour (cervix 17 cm--fully dilated). Fetal cotyledons and membranes had very low oxytocin receptor concentrations during most of pregnancy, on average only 20 fmol mg-1 protein. At term and during labour, receptor concentrations were significantly increased in both tissues. Mean concentrations during labour were 163 +/- 36 fmol mg-1 for cotyledons, 270 +/- 61 fmol mg-1 for chorioallantois and 311 +/- 121 fmol mg-1 for amnion.(ABSTRACT TRUNCATED AT 250 WORDS)


Inhibitory effects of a luteinizing-hormone-releasing hormone agonist implant on ovine fetal gonadotrophin secretion and pituitary sensitivity to luteinizing-hormone-releasing hormone. Sheep fetuses at day 70 of gestation (term = 145 days) were implanted subcutaneously with a biodegradable implant containing a luteinizing-hormone-releasing hormone (LHRH) agonist (buserelin) to investigate whether treatment with LHRH agonist would induce a state of desensitization of the fetal gonadotrophs and thus influence fetal gonadal development. Treatment with the LHRH agonist for 35-40 days caused a significant reduction in mean fetal plasma concentrations of LH and follicle-stimulating hormone (FSH) compared with control fetuses. LH pulses were evident in control fetuses but were completely abolished by buserelin treatment. Furthermore, the pituitary content of LH and FSH was significantly depleted in fetuses implanted with LHRH agonist. A bolus intravenous injection of 500 ng LHRH given to control fetuses caused a rapid and significant increase in plasma LH and FSH concentrations which was sustained for at least 60 min after injection. Pretreatment with buserelin completely abolished the LH and FSH responses to a bolus injection of LHRH. There were no differences between the sexes in fetal gonadotrophin concentrations or pituitary sensitivity to LHRH in control or agonist-treated fetuses. Furthermore, buserelin treatment for 35-40 days had no effect on the morphological appearance of the fetal gonads when compared with control fetuses, at least to day 110 of pregnancy. These results provide evidence for the induction of a state of desensitization of the LHRH receptors of the fetal pituitary gonadotrophs following long-term treatment with an LHRH agonist, but provide no evidence for a role for gonadotrophin secretion in gonadal development at this stage in fetal life.


Successful cryopreservation of mouse blastocysts using a new vitrification solution. Mouse blastocysts were exposed to solutions containing four concentrations (10, 20, 30 and 40% v/v) of six permeating cryoprotectants (glycerol, ethylene glycol, propylene glycol, dimethyl sulfoxide, 1,3-butanediol and 2,3-butanediol) in phosphate-buffered saline (PBS) with calf serum (CS) at room temperature (20-22 degrees C). Blastocysts were exposed to these solutions for various periods, diluted into PBS plus CS with or without 1 mol trehalose l-1 solution and their subsequent survival in vitro was examined. Two-way anova showed a significant interaction (P < 0.01) between cryoprotectant type, concentration of cryoprotectant and method of dilution. However, no significant interaction was observed between cryoprotectant type and duration of exposure. Results suggest that cryoprotectant-induced injury to nonfrozen blastocysts is variable and depends on the cryoprotectant used. On the basis of toxicity assays, ethylene glycol was the least harmful and was combined with dimethyl sulfoxide and 1,3-butanediol to produce a new vitrification solution. Mouse blastocysts were successfully cryopreserved using a vitrification solution (designated as VSv) consisting of 20% ethylene glycol, 20% dimethyl sulfoxide and 10% 1,3-butanediol (v/v). Embryos were equilibrated in two steps, first in an equilibration solution (designated as ESv: 10% ethylene glycol, 10% dimethyl sulfoxide and 5% 1,3-butanediol; v/v) and then to VSv or one-step in VSv at different exposure times at room temperature, and then vitrified by direct plunging into liquid nitrogen. High developmental rates were obtained in vitro when the embryos were exposed to ESv and VSv for 3 and 0.5 min, respectively (96.2%) or exposed to VSv for 0.5 min (95.4%). Prolonged exposure time proved detrimental to subsequent embryo development in vitro. When vitrified warmed embryos were transferred immediately to pseudopregnant recipients, the rate of development to normal fetuses did not significantly differ from that of the nonvitrified control (two-step, 54.2 and one-step, 45.0 versus 60.0%, P > 0.05). These results suggest that the simple vitrification solution described in this study is effective for the cryopreservation of mouse blastocysts.


International collaborative study by in vitro bioassays of the first international standard for porcine inhibin. A lyophilized preparation of inhibin from porcine ovarian follicular fluid, ampoule code 86/690, was made internationally available as a research standard for in vitro bioassays in 1987. A study involving ten participants in eight countries assessed the stability and suitability of this research standard to serve as an international standard. Each of the participants used in vitro assays, the majority of which depended upon the inhibition of release of follicle-stimulating hormone from dispersed rat anterior pituitary cells. The research standard 86/690 was compared with coded ampoules of 86/690 stored under conditions of accelerated thermal degradation and with inhibins from different species. Intra- and interlaboratory variation for estimates of potency of a coded duplicate ampoule of the research standard provided the basis for comparisons of non-identical inhibins, but the fourfold variability of potency estimates for identical ampoules was such that no conclusions about the differences seen for non-identical inhibins could be made. Predictions of stability from consensus estimates of potency of ampoules that have undergone accelerated thermal degradation indicated that the research standard had satisfactory stability. On the basis of this study, the research standard 86/690 was deemed sufficiently stable and suitable to serve as a standard for in vitro bioassays and was established by the World Health Organization Expert Committee on Biological Standardization as the First International Standard for Porcine Inhibin. The possible presence, in biological extracts (standard or sample), of other bioactive proteins, such as activin and follistatin, complicates the quantitative interpretation of bioassay data. A standard of highly purified human inhibin is now required as a standard for immunoassays used for clinical research purposes; sufficient quantities of recombinant human inhibins have recently been donated for ampouling and evaluation by bio- and immunoassay in the subsequent phase of the standardization of inhibins.


Possible implication of lysophosphatidylcholine in cell fusion accompanying implantation in rabbits. Implantation in rabbits involves the cellular fusion of trophoblastic and uterine epithelial cells resulting in embryo penetration of the uterine endometrium. Since lysophospholipids, known to have fusigenic properties, could be responsible for this cell fusion, the metabolism of lysophospholipids was studied throughout gestation in blastocyst/yolk sac and extracoelic amnioallantoic fluids. Analysis of phospholipid composition revealed that lysophospholipids are present in blastocyst/yolk sac fluid. Their concentrations and haemolytic activity change during pregnancy. They increase and reach their highest values during days 7 to 9, the implantation days in rabbits. A clear correlation was observed between lysophosphatidylcholine concentrations in blastocyst/yolk sac fluid and haemolysis induced by this fluid. Phosphatidylcholine concentrations, phospholipase A2 activity, which generates lysophospholipids, and lysophospholipase A activity which hydrolyses lysophosphatidylcholine into fatty acid, were at their highest value at day 12. These data suggest that a transient accumulation of lysophospholipids could ensure local cell fusion. Moreover, we propose that the lysophospholipid concentrations in blastocyst/yolk sac fluid are dependent upon activities of phospholipase A2 and lysophospholipase.


Ovarian and pituitary function in dogs after hysterectomy. In studies of five hysterectomized and five control dogs, hysterectomy shortened the anoestrous interval (96.6 +/- 28.0 versus 149.4 +/- 50.9 days, P < 0.05). No differences in hormone concentrations (progesterone, oestradiol, prolactin and growth hormone) were observed between the control and hysterectomized dogs except for a brief fall in progesterone concentrations over 8 days immediately after surgery, between days 35 and 40 after onset of pro-oestrous bleeding; only these animals developed symptoms of overt pseudopregnancy. It is concluded that, in dogs, luteal regression occurs independently of a uterine luteolysin, but that the uterus may play a role in control of duration of anoestrus. Pseudopregnancy seems to be initiated by a fall in progesterone concentrations rather than by other hormonal changes.


Effects of naloxone on circulating gonadotrophin concentrations in prepubertal heifers. The pattern and opioidergic control of the secretion of gonadotrophins in prepubertal heifer calves were examined. Ten age-matched Hereford heifer calves were weighed and a blood sample was taken every 2 weeks from 2 to 25 weeks of age and then weekly until 60 weeks of age. At 60 weeks, a fertile bull was introduced and at 75 weeks of age pregnancy diagnosis was performed by transrectal ultrasonography. At 4, 12, 18, 24 and 32 weeks of age, the opioid antagonist naloxone was injected (i.v., n = 5; 1 mg kg-1 body weight) each hour for 12 h. Control heifers received sterile saline at the same ages. Blood samples were collected every 12 min for the 12 h treatment and serum samples were analysed for luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Samples taken once every 2 weeks from 2 to 60 weeks were analysed for LH, FSH and oestradiol, and weekly samples were taken for progesterone determination. There was no effect of naloxone on the age at puberty, which was 56.2 +/- 0.7 weeks at a body weight of 388.5 +/- 8.0 kg. The mean age at conception was 63.4 +/- 0.5 weeks. On the basis of samples taken every other week, serum concentrations of LH were high at 10 weeks and between 40 and 60 weeks of age. From the periods of intensive blood collection, the early rise in mean serum concentrations of LH appeared later at 12 and 18 weeks of age and was caused by a rise in LH pulse amplitude.(ABSTRACT TRUNCATED AT 250 WORDS)


Cyclic adenosine 3'5'-monophosphate and the relaxant action of relaxin in the rat uterus in vivo. The relationship between relaxant actions of relaxin in the uterus and changes in uterine cAMP concentrations was assessed in anaesthetized bilaterally ovariectomized nonpregnant rats. Relaxin i.v. bolus (5 micrograms kg-1) did not change cAMP concentrations but inhibited uterine contractions with rapid onset. Uterine contractions were significantly reduced by 30-70% for 60 min. Relaxin (50 micrograms kg-1) produced a short-lived (up to 5 min) and small (up to 3.2-fold) increase in cAMP concentrations plus a marked (90%) and prolonged inhibition of uterine contractions (70-90% over 60 min). Salbutamol (an agonist at beta 2-adrenoceptors, 100 and 500 micrograms kg-1) produced a similar degree and time course of inhibition of uterine contractions to that of relaxin but a more marked (19-fold) increase in cAMP concentrations. Glibenclamide, a blocker of ATP-sensitive potassium channels, which has been shown to antagonize relaxin as a uterine relaxant, did not prevent the relaxin-induced rise in cAMP concentrations. It is suggested that the uterine relaxant action of relaxin may not result from an increase in uterine cAMP concentrations.


Suppression of pulsatile luteinizing hormone secretion by gonadotrophin-releasing hormone antagonist does not affect episodic progesterone secretion or corpus luteum function in ewes. Progesterone secretion has been observed to be episodic in the late luteal phase of the oestrous cycle of ewes and is apparently independent of luteinizing hormone (LH). This study investigated the effects of suppressing the pulsatile release of LH in the early or late luteal phase on the episodic secretion of progesterone. Six Scottish Blackface ewes were treated i.m. with 1 mg kg-1 body weight of a potent gonadotrophin-releasing hormone (GnRH) antagonist on either day 4 or day 11 of the luteal phase. Six ewes received saline at each time and acted as controls. Serial blood samples were collected at 10 or 15 min intervals between 0 and 8 h, 24 and 32 h, and 48 and 56 h after GnRH antagonist treatment and daily from oestrus (day 0) of the treatment cycle for 22 days. Oestrous behaviour was determined using a vasectomized ram present throughout the experiment. Progesterone secretion was episodic in both the early and late luteal phase with a frequency of between 1.6 and 3.2 pulses in 8 h. The GnRH antagonist abolished the pulsatile secretion and suppressed the basal concentrations of LH for at least 3 days after treatment. This suppression of LH, in either the early or late luteal phase, did not affect the episodic release of progesterone. Daily concentrations of progesterone in plasma showed a minimal reduction on days 11 to 14 after GnRH antagonist treatment on day 4, although this was significant (P < 0.05) only on days 11 and 13. There was no effect of treatment on day 11 on daily progesterone concentration, and the timing of luteolysis and the duration of corpus luteum function was unaffected by GnRH antagonist treatment on either day 4 or day 11. These results indicate that the episodic secretion of progesterone during the luteal phase of the oestrous cycle in ewes is independent of LH pulses and normal progesterone secretion by the corpus luteum can be maintained with minimal basal concentrations of LH.


[Famotidine in the treatment of stomach ulcer]. In order to evaluate the efficacity and tolerance of Famotidine in the treatment of gastric ulcer, in Lebanese patients, we conducted a prospective open study in twenty one patients with benign gastric ulcer. A clinical, endoscopic and biological evaluation was done before, at 4 weeks and if necessary at 8 weeks after the start of 40 mg of Famotidine per day. All patients were carefully monitored at regular intervals for adverse drug reactions by clinical and biological examinations. The healing rate at 4 and 8 weeks was 75% and 91% respectively. The treatment was well tolerated and no modifications in laboratory tests were observed. Famotidine therefore proved effective in the treatment of gastric ulcer and was well tolerated on a short term basis.


[Various aspects of uretero-ureteral reflux in incomplete ureteral duplication]. This study is a complete review of the common complications related to incomplete ureteral duplication, and based on three clinical cases. These complications include: Recurrent urinary tract infections with pyelonephritis; non functioning of one or both kidney units; Urinary lithiasis and the uretero-ureteral reflux secondary to retrograde ureteral peristalsis. Finally, the surgical treatment of the uretero-ureteral reflux depends on the clinical and radiological symptoms of the patients. Heminephrectomy in case of a non functioning renal segment. Excision of one ureteral segment converting an incomplete ureteral duplication into a bifid pelvis. Conversion of an incomplete duplication into a complete ureteral duplication if the common segment is near the bladder. Nephroureterectomy should be considered in case of non functioning kidney with type IV vesico-ureteral reflux.


[The estimation of the prevalence of diabetes mellitus in Lebanon]. A sample of 436 persons of both sexes, all ages and being from all over the Lebanese territory, was analysed as part of a survey on the prevention of cardiovascular diseases. Our results gave an estimation of the prevalence of diabetes mellitus in Lebanon being at 7-8% and that of impaired glucose tolerance being at 10-11%. 50% of diabetics were previously unknown; this percentage rose up to 89% in people aged less than 40 years. The male preponderance was not statistically significant. The prevalence of diabetes mellitus increased steadily with age.


Gonadotropin secreting pituitary microadenoma. FSH and LH secreting pituitary adenomas may not be as rare as has been previously thought. Women in their reproductive years with a normal estrogenic state and hypergonadotropic amenorrhea should be suspected to harbor a pituitary microadenoma secreting gonadotropins. A computerized tomogram of the pituitary confirms the diagnosis. Two such cases who underwent transphenoidal resection of their adenomas are presented. Pituitary dynamic testing is normal in both. Laparoscopic directed ovarian biopsies show primordial follicles in one and a corpus luteum in the other. Transphenoidal resection, with the risk of permanent hypopituitarism, should be weighed against expectant management, with periodic evaluation of pituitary functions, until the natural history of these adenomas is uncovered.


Osteosarcoma clinical and laboratory evaluation with staging. An outline of the clinical presentation, laboratory findings and staging of osteosarcoma is discussed. Experimental studies that may help in the future in the diagnosis are also mentioned.


Outline of conventional management of osteosarcoma. Major change in the prognosis of osteosarcoma has been made over the past two decades. The survival rate is over 80%, each case must be investigated by modern methods and classified according to the Enneking classification. Biopsy techniques must be very precise. Chemotherapy could be administered prior to surgery and modified according to the response to the agent used. It could be also given post operatively. Surgery of the extremity should be conservative when the local and general condition is permissible.


[The cancer registry at the Hôtel Dieu de France Hospital]. Between January and December 1989, 541 new cases of cancer have been diagnosed at Hôtel-Dieu de France Hospital at Beirut. These cases were among 5400 histopathologic or hemato-cytologic examinations performed during the same period. There were 311 men (57.5%) and 230 women (42.5%). In men lung cancer was the most common site (19%) followed by bladder cancer (16.7%) and prostate cancer (11.6%). In women the most frequent cancer reported was breast cancer (36.1%) followed by uterine (body and cervix) cancer (15.2%) and digestive tract tumors (12.3%). Hematologic malignancies were more frequent between children and young patients. We have noted a progressive frequency of breast, lung, digestive tract, bladder and genito-urinary cancers with age. The establishment of a national tumor registry covering all diagnosed patients in different Lebanese Institutions is warranted.


[How to improve the results in colorectal cancer?]. To improve results in the treatment of colo-rectal cancer, there are four main ways to consider: 1--Earlier diagnosis even presymptomatically (Screening). 2--More radical operations. 3--Adjuvant radiotherapy and chemotherapy in selected cases. 4--More strict follow-up with CEA monitoring and occasional "second look operations".


[Gastric leiomyoblastoma, 2 case reports with a selective literature review]. The gastric Leiomyoblastoma is a smooth muscle described in 1960 by Martin and in 1962 by Stout. These rare tumors are generally benign but may have a sarcomatous evolution in 11% of cases. They are characterized by the presence of large round or polygonal cells. Thin filaments are visible on electronic microscopy. Treatment of choice is the surgical resection.


[The use of antiemetics in cancer chemotherapy]. The development of new chemotherapeutic protocols especially those cisplatin containing regimens, has lead to a dramatic improvement of remission rates in certain cancers. Nausea and vomiting seems to be the most distressing side effect from the patients point of view. A good administration of classical anti-emetic drugs should yield to at least 50% of good protection. The introduction of new drugs should even give more protection to our patients.


[Use of implantable chemotherapy reservoirs. Experience at the Hotel-Dieu of France]. We are presenting our experience with 70 deep venous catheters for long term chemotherapy. Despite the well known complications of these procedures, the implantation of subcutaneous injection sites has facilitated the management and brought much more comfort to cancer patients. Strict measures of survey and meticulous nursing care are essential for optimal use of the injection sites.


[Vaginal delivery with scarred uterus at the Hotel-Dieu of France, Beirut]. Vaginal deliveries on a scarred uterus are actually admitted and done in many obstetrics-gynecology centers throughout the world. A uterine scar does not represent a formal indication for a repeat cesarean section anymore. In this article, the results of a retrospective study between January 1980 and December 1989 are reported. We reviewed all the files of women having a scarred uterus, delivering at "Hotel-Dieu de France", vaginally or by repeat cesarean section. This study concerns 560 women: 500 delivering by repeat cesarean section & 60 having a trial of labor, 45 of which delivering vaginally. A lot of parameters are compared, such as: days of hospitalization, antibiotic coverage, and materno-foetal complications. No uterine rupture was reported. The hospital stay was reduced following vaginal delivery after the trial of labor. A trial of labor on a scarred uterus, taking some precautions, seems tempting, without enhancing feto-maternal morbidity and mortality.


[Inguinal herniorrhaphies by the pre-peritoneal method using surgical mesh]. During a period of 5 years, 314 patients with 427 inguinal hernias were operated. 80 males with bilateral inguinal hernia recurrent, or not, were operated with an insertion of a Crinoplaque by a pre-peritoneal approach. The operative time was 65 +/- 26 minutes. 10 patients (12.5%) had by the same incision a concomitant pelvic procedure (9 varicoceles, 1 benign prostatic hypertrophy). 4 patients (5%) developed minor parietal complications. After 1 to 4 years of follow-up, 1 patient (1,25%) showed one side recurrence, in the beginning of our experience. In conclusion, we consider the pre-peritoneal Crinoplaque insertion as a treatment of choice for bilateral recurrent, or not, inguinal hernia for its simplicity and its lower complication rate.


Laparoscopic cholecystectomy: initial experience in Lebanon. Laparoscopic Cholecystectomy became very rapidly the procedure of choice for dealing with gallstones. We present our initial experience in Lebanon with 45 cases. All patients referred for cholelithiasis without any exception had a laparoscopic procedure. The conversion, mortality and morbidity rates were 0%. Post-operative hospital stay was 24 hours in the majority of the cases. The results clearly demonstrates the major advantages this procedure has over the conventional cholecystectomy and we believe it should be the procedure of choice for cholelithiasis.