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Biomedical subjects

A Locatelli

Publications and source records attributed to A Locatelli.

At least 91 records · Page 5Linked to original sources

Effects of gonadotrophin deprivation on follicular growth in gilts.

The endocrine and ovarian effects of hypophysectomy (n = 5) and gonadotrophin-releasing hormone (GnRH) antagonist administration (Antarelix) (n = 5) were studied in gilts by comparison with control animals (n = 6). All gilts were given Regumate (20 mg/d for 18 d). The last day of Regumate was day 0. Hypophysectomy and initiation of Antarelix administration (0.6 mg iv twice daily for 7 d) were performed on day 5. All ovaries were obtained at slaughter on day 12. Blood samples were obtained daily from all Antarelix-treated and control sows to measure luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations. Frequent sampling was performed on day 10 on control and Antarelix-treated gilts to assess pulsatile LH secretion. Oestrus or LH surge was initiated before the beginning of treatment in 2 hypophysectomized, 4 Antarelix-treated, and 4 control females. Gonadotropins were undetectable in the blood of hypophysectomized sows on day 6. On day 10, pulsatile LH release was blocked in Antarelix-treated gilts. At no time were FSH concentrations significantly affected. Histological observation of the ovaries demonstrated that: (i) similar populations of healthy or total (healthy + atretic) follicles < 1 mm were found in the 3 groups of females; (ii) healthy follicles 1 to 2 mm in diameter were present in Antarelix-treated but not in hypophysectomized gilts; and (iii) healthy follicles > 2 mm were absent in Antarelix-treated and hypophysectomized gilts. The present study suggests the existence of 3 subgroups amongst antral follicles (gonadotropin independent: 0.19 to 1.1 mm; FSH dependent: 1.1 to 2 mm; LH pulses dependent: > 2 mm in diameter).

Animals↗

Intrauterine diagnosis and management of transient myeloproliferative disorder.

Transient myeloproliferative disorders can be associated with hydrops in Down syndrome fetuses. No cases of prenatal management of such a condition have been reported in the literature. We report a case of myeloproliferative disorder diagnosed by cordocentesis at 31 weeks in a Down syndrome fetus with pericardial effusion. A pericardiocentesis was performed at the first signs of hydrops and successfully improved fetal cardiac function, allowing for continuation of pregnancy.

Adult↗

Regulation of luteinizing hormone-releasing hormone (LHRH) secretion by melatonin in the ewe. I. Simultaneous delayed increase in LHRH and luteinizing hormone pulsatile secretion.

This study was performed to test the hypothesis that a short-day-like melatonin treatment would induce simultaneous stimulations of LHRH and LH secretion. Simultaneous serial samples of jugular and portal blood to assay LH and LHRH, respectively, were carried out at three different periods according to the expected changes in LH secretion. Twenty-six ovariectomized ewes bearing a subcutaneous implant of estradiol and exposed to a long-day photoperiod (16L:8D) received s.c. implants of melatonin on Day 0. LHRH and LH pulsatile secretion was studied in the first group during inhibition of LH secretion by long days (on Day -1, n = 6). A second group (n = 9) was sampled on Day 39. Moreover, seven animals from this group were sampled on Day 46 to focus on the beginning of LH increase. The third group (n = 6) was sampled during maximal LH secretion (Day 74, n = 6). A fourth group (n = 5) composed of noncannulated animals was sampled for LH secretion only as a control for the effects of surgical procedures. On each occasion, blood was collected every 10 min for 6 h. Between Day -1 and Day 39, LHRH pulse frequency was low and did not differ. In the same animals, this value became 2.5-fold higher between Day 39 and Day 46. Finally, on Day 74, LHRH pulse frequency was about 6-fold higher than on Day -1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Regulation of luteinizing hormone-releasing hormone (LHRH) secretion by melatonin in the ewe. II. Changes in N-methyl-D,L-aspartic acid-induced LHRH release during the stimulation of luteinizing hormone secretion by melatonin.

Injection of N-methyl-D,L-aspartic acid (NMDA), a glutamatergic agonist, cause an acute release of LH. The amount of LH released is modulated by photoperiod in sheep; it is larger in animals in which reproductive activity has been inhibited by long days than in those photostimulated by short days. The purpose of the present experiment was to determine whether this effect of photoperiod on LH response to NMDA is mediated by melatonin and whether it reflects a change in the amount of LHRH released in response to NMDA. Twenty-one ovariectomized ewes, bearing an s.c. implant of estradiol and kept in long days (16L:8D), received s.c. implants of melatonin on Day 0. The effect of a single injection of NMDA (5 mg/kg i.v.) on LHRH and simultaneous LH secretion was studied at three different periods of the stimulation of LH secretion by the melatonin treatment: first, during inhibition of LH secretion by long days (on Day -1, n = 6); secondly, during the onset of the LH increase (on Day 39, n = 9); and thirdly, during maximum LH secretion (on Day 74, n = 6). The stimulation of LHRH secretion by administration of NMDA was characterized by a large and long increase in LHRH concentration on Day -1 and Day 39. There was no significant difference between these two periods in terms of duration and amplitude of the LHRH release. On Day 74, the stimulation was much shorter than on Day -1 and Day 39 and was followed by a phase of decreased pulsatility of LHRH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Bilateral lesions of the suprachiasmatic nuclei alter the nocturnal melatonin secretion in sheep.

The hypothalamic suprachiasmatic nuclei (SCN) constitute both the biological clock of many circadian rhythms, and the first relay in the transmission of light cues from the retina to the pineal gland, which releases, via nocturnal melatonin secretion, an endocrine expression of the daylength. The aim of the present work was to investigate the precise role of the SCN in the entrainment of the nocturnal rhythm of melatonin (MEL) in sheep. Bilateral lesions of the SCN were performed via a transsinusal surgical approach in 10 adult rams submitted to a constant photoperiod (16:8D). Lesioned rams were compared to 4 sham and 2 control animals. Blood samples were collected 8 days before, 8 days after, and one month after surgery. Plasma MEL levels were estimated using direct radioimmunoassay. At the end of the experiment, histology and immunohistochemistry of the suprachiasmatic area were performed, and the extent of lesions was evaluated using a computerized image analysis system. Six rams exhibited a complete lesion of the SCN, and in the four remaining animals, the lesions were restricted to the anterior part of the SCN. For all animals, the nocturnal melatonin secretion was altered, but depending on the extent of the lesion, two types of results were observed: Eight days after surgery, in 3 of the 4 rams bearing anterior lesions of the SCN (SCNx-), a nocturnal increase in melatonin secretion still occurred at dusk, but the duration of this secretion extended beyond the end of the night. One month after surgery, melatonin profiles were once again normal, as compared to sham animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Nature and bioactivity of gonadotropin-releasing hormone (GnRH) secreted during the GnRH surge.

Previous studies have demonstrated a neural action of estradiol in inducing a surge of GnRH in the ewe. However, although the GnRH and LH surges began concurrently, the GnRH surge consistently continued well beyond the surge of LH. Three experiments were conducted to test the hypothesis that the termination of the LH surge results from the secretion of a relatively inactive variant of GnRH during the later phases of the GnRH surge. In the first experiment, hypophyseal portal blood collected during an estrogen-induced LH surge was analyzed for GnRH immunoreactivity using two antibodies having specificity for the N- or C-terminal portion of the GnRH molecule. The duration, amplitude, and time course of the GnRH surge were found to be similar irrespective of the antisera used. In a second experiment, a competitive GnRH antagonist was administered at the beginning of the estrogen-induced GnRH/LH surge at a dose capable of blocking pituitary responsiveness for approximately half the duration of the GnRH surge. Antagonist treatment did not result in any change in the time of onset of the GnRH surge, but there was no increase in LH that naturally occurs coincident with onset of the GnRH surge. Rather, a persistent increase in LH secretion was observed during the latter stages of the GnRH surge, indicating that the GnRH molecules secreted at this time were biologically active. Finally, a sensitive and specific ovine pituitary cell bioassay was used to test bioactivity of GnRH in hypophyseal portal blood during different phases of the GnRH surge. GnRH bioactivity in samples collected early in the GnRH surge was greater than that before the onset of the GnRH surge but no greater than that collected during the descending limb of the surge. The results of all three experiments fail to support the hypothesis that the LH surge ends because of a change in the nature of the GnRH secreted. Rather they show that GnRH secreted throughout the surge is biologically active. Thus, the termination of the LH surge before that of the GnRH surge occurs for reasons other than lack of a bioactive GnRH signal.

Animals↗

Ontogenesis of somatic and germ cells in sheep fetal testis.

Testicular development of sheep fetuses was studied between day 42 of gestation and birth. Testis mass and the total number of testicular cells increased curvilinearly with fetal age and a positive linear relationship was established between the logarithmic values of age and testis mass, sex cord total length, total number of Sertoli cells, germ cells and Leydig cells per testis. The mean number of gonocytes per unit length of sex cord, the Sertoli cell nuclear cross-sectional area and the Leydig cell cross-sectional area decreased linearly with age between day 42 of gestation and birth. Hypophysectomy and hemicastration were performed to study the regulation of testicular cell divisions during fetal life and to determine whether they were under pituitary control and whether a feedback mechanism was present. Hypophysectomy at day 100 or 110 of gestation nonsignificantly decreased (0.05 < P < 0.01) the testis mass, total length of sex cords and total number of Sertoli cells and significantly decreased (P < 0.05) the cross-sectional area of Leydig cells and nuclei of Sertoli cells. Sex cord diameter and total number of gonocytes were unaltered. Hemicastration at day 110 of gestation significantly increased (P < 0.05) the total number of Leydig cells per testis without changing any other testicular parameter. In male sheep fetuses, the proliferation of testicular somatic and germ cells occurs throughout testicular fetal growth at a higher rate before day 100 of gestation than later, but without any differentiation. Mitotic divisions of Sertoli cells are more numerous before birth than afterwards. Before birth, the proliferation of gonocytes is not under pituitary control.

Animals↗

[Quantification and monitoring of vascular resistance in the lower limbs by the Doppler method (animal model)].

The object of this study was to define and validate a non-invasive method of evaluation and monitoring of vascular resistances in the leg. Blood flow velocity was measured by Doppler ultrasound in an animal model (ewe) with similar blood flow characteristics in the lower limb as man and allowing access to the required invasive measurements for validation of the method (pressure and flow). Vascular resistances distal to the measuring point (femoral, for example) were assessed using the resistance index R = D/S, S being the peak systolic deflection and D that of diastolic reflux of the Doppler spectral analysis of flow in the femoral artery. The values and variations of this resistance index were compared with the vascular resistances calculated from measurements of pressure and flow at the point of Doppler sampling and expressed in mmHg/ml/min. Femoral flow was measured by Doppler ultrasound (Doppler-echo), and mean pressure by an arterial catheter introduced into the abdominal aorta. Compression of the lower limb veins induced a venous return resulting in a reduction of cardiac output and femoral flow. During compression, femoral flow decreased by an average of 29% (p < 0.001) although mean pressure and heart rate did not change significantly. The femoral resistance index (Rf) increased by an average of 37.5% (p < 0.01) and vascular resistances increased by 45.9% (p < 0.01). Injection of 1 mg adrenaline induced peripheral vasoconstriction with an increase in blood pressure and a decrease in heart rate and femoral flow.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Resistance to aminoglycosides and other antibiotics among clinical isolates of Enterococcus spp.

We conducted a study on 81 clinical isolates of enterococci (65 Enterococcus faecalis and 16 Enterococcus faecium) collected consecutively over a one year period at our University Hospital. Of them, 10 (15%) Enterococcus faecalis and 6 (38%) Enterococcus faecium showed high level resistance to aminoglycosides, were all acquired in surgical units and in five cases they were responsible for invasive infections. Ampicillin and imipenem were active only against Enterococcus faecalis. Vancomycin and teicoplanin were bacteriostatic; however, teicoplanin MICs were lower and in 45% of cases its MBC values were within clinically achievable serum concentrations. Ciprofloxacin and cotrimoxazole were active in vitro.

Aminoglycosides↗

Treatment of recurrent and resistant CAPD peritonitis by temporary withdrawal of peritoneal dialysis without removal of the catheter.

Recurrent and resistant continuous ambulatory peritoneal dialysis (CAPD) peritonitis is usually treated by removal of the catheter and temporary hemodialysis. We treated 3 patients: 1 with resistant Klebsiella peritonitis and 2 with recurrent peritonitis (one due to Staphylococcus and the other to Enterococcus), by stopping CAPD for a 2-4 week period, leaving the catheter in situ and continuing antibiotic therapy. All 3 patients had resolution of their infections and restarted CAPD. This therapeutic modality reduced catheter replacements, limited admissions to the hospital, reduced psychological impact, and diminished risks and costs of CAPD.

Adult↗

Density of peptide histidine-isoleucine- and vasoactive intestinal peptide-immunoreactive nerve fibers in the sheep pineal gland is not affected by superior cervical ganglionectomy.

Peptide histidine-isoleucine (PHI) is a regulatory peptide, synthesized as part of the same propeptide that includes also vasoactive intestinal peptide (VIP). The present study describes the distribution of PHI-immunoreactive nerve fibers in the sheep pineal organ and compares their location with the distribution of VIP-immunoreactive fibers in both normal and superior cervical ganglionectomized sheep in order to elucidate the origin of the PHI/VIP immunoreactive nerve fibers. Several PHI-immunoreactive nerve fibers were present in the meninges and in the pineal capsule. Numerous positive nerve fibers entered the pineal gland and travelled within connective tissue spaces. Individual PHI-positive nerve fibers were either smooth, without specialization, or varicose. Generally VIP- and PHI-immunoreactive fibers were located close to connective septa and blood vessels. However, many PHIergic and VIPergic fibers possessing varicosities of variable sizes were also dispersed between pinealocytes. The distribution, density, and morphology of PHI- and VIP-immunoreactive fibers in the sheep pineal gland were similar. In superior cervical ganglionectomized animals, intrapineal VIP- and PHI-immunoreactive nerve fibers were present with the same density as in control animals. In agreement, the concentration of immunoreactive VIP and PHI did not change after ganglionectomy. No VIP- and PHI-immunoreactive cell bodies were observed in the superior cervical ganglia. Thus this study shows that the intrapineal VIP- and PHI-immunoreactive nerve fibers do not originate from the sympathetic superior cervical ganglion.

Animals↗

The suprachiasmatic nucleus in the sheep: retinal projections and cytoarchitectural organization.

The retinal innervation, cytoarchitectural, and immunohistochemical organization of the suprachiasmatic nucleus (SCN) was studied in the domestic sheep. The SCN is a large elongated nucleus extending rostrocaudally for roughly 3 mm in the hypothalamus. The morphology is unusual in that the rostral part of the nucleus extends out of the main mass of the hypothalamus onto the dorsal aspect of the optic chiasm. Following intraocular injection of wheat-germ agglutinin-horseradish peroxidase or tritiated amino acids, anterograde label is distributed throughout the SCN. Retinal innervation of the SCN is bilaterally symmetric or predominantly ipsilateral. Quantitative image analysis demonstrates that, although the amount of autoradiographic label is greatest in the ventral and central parts of the nucleus, density varies progressively between different regions. In addition to the SCN, retinal fibers are also seen in the medial preoptic area, the anterior and lateral hypothalamic area, the dorsomedial hypothalamus, the retrochiasmatic area, and the basal telencephalon. Whereas the SCN can be identified using several techniques, complete delineation of the nucleus requires combined tract tracing, cytoarchitectural, and histochemical criteria. Compared with the surrounding hypothalamic regions, the SCN contains smaller, more densely packed neurons, and is largely devoid of myelinated fibers. Cell soma sizes are smaller in the ventral SCN than in the dorsal or lateral parts, but an obvious regional transition is lacking. Using Nissl, myelin, acetylcholinesterase, and cytochrome oxidase staining, the SCN can be clearly distinguished in the rostral and medial regions, but is less differentiated toward the caudal pole. Immunohistochemical demonstration of several neuropeptides shows that the neurochemical organization of the sheep SCN is heterogeneous, but that it lacks a distinct compartmental organization. Populations of different neuropeptide-containing cells are found throughout the nucleus, although perikarya positive for vasoactive intestinal polypeptide and fibers labeled for methionine-enkephalin are predominant ventrally; neurophysin-immunoreactive cells are more prominent in the dorsal region and toward the caudal pole. The results suggest that the intrinsic organization of the sheep SCN is characterized by gradual regional transitions between different zones.

Acetylcholinesterase↗

Thienylvinylindoles as inhibitors of mitochondrial NADH dehydrogenase.

In connection with a previous study, new phenylindoles bearing a 2- or 3-thienyl group were synthesized and tested as specific inhibitors of mitochondrial NADH dehydrogenase. The position of the phenyl ring and the geometrical configuration play an important role in the activity and specificity of these derivatives. In order to study the mechanism of action of these thienylvinylindoles, their activity was compared with that of known inhibitors in a new test employing exogenous quinones.

Animals↗

A new surgical approach to the ram suprachiasmatic nuclei.

In mammals, lesions of the suprachiasmatic nuclei (SCN) cause alterations in several biological rhythms. Investigation of the role of the ovine SCN requires complete bilateral lesions of the nuclei. However, the elongated shape, location and horizontal orientation of this hypothalamic structure prohibit a classical stereotaxic vertical approach. We present here a new trans-sinusal surgical technique to reach the SCN for lesion or other studies, which avoids damage to hypothalamic or other structures. The surgical procedure allows direct viewing of the SCN, located dorsal to the optic chiasm on the rostro-ventral surface of the brain. The orientation of the access tunnel is roughly aligned with the nearly horizontal main axis of the nuclei. An example of the use of such a surgical procedure is described: the bilateral destruction of the SCN. Following surgery, the extent of lesions was assessed by histology, showing complete destruction of the SCN. In order to demonstrate the physiological effects of the lesions, melatonin secretion profiles were followed over 48 h periods and compared before and after surgery. Bilateral SCN lesions induced alterations in the rhythm of melatonin secretion: levels were no longer correlated with light/dark cycles.

Animals↗