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Biomedical subjects

C Orlando

Publications and source records attributed to C Orlando.

At least 73 records · Page 4Linked to original sources
PubMed1992

Sertoli-germ cells interactions in the human testis.

In previous histoimmunochemical studies we reported that transferrin (TF) and insulin-like growth factor I (IGF-I) are present in the cytoplasm of the Sertoli cells of the adult human testis. Receptors for TF were found mainly in adluminal germ cells and type I receptors for IGF-I both in Sertoli and germ cells. Using electron microscopy, evidence of transfer of both TF and IGF-I from the Sertoli to the germ cells through a receptor-mediated endocytosis mechanism was also found. In this paper we report the results of the histoimmunochemical localization of alpha inhibin in the human fetal, prepubertal and adult testis. In 8- to 14-week-old fetal testes a positive immunostaining was found mainly in the interstitial cells, whereas no staining was found in the germ cords. In the prepubertal testis the immunostaining was present in the Sertoli cells but not in the interstitial cells. In the adult human testis the immunostaining was present not only in the Sertoli cells but also in the spermatocytes and in several Leydig cells. Using electron microscopy and immunogold labeling the presence of alpha inhibin immunoreactivity was found in the rough endoplasmic reticulum and in the Golgi cisternae of both Sertoli and Leydig cells. Moreover we found evidence of transfer of alpha inhibin from the Sertoli to the germ cells through receptor-mediated endocytosis.

Adult↗
PubMed1992

Endothelin in rabbit uterus during pregnancy.

Specific immunostaining for endothelin-1 (ET-1) was observed in the uterine mucosa of pregnant and nonpregnant rabbits. During gestation, giant cells immunopositive for ET-1 were identified in either endometrium or myometrium. In the latter, they were located in close proximity to the muscular fibers. High-affinity (dissociation constant = 0.25 nM) high-capacity [maximal binding (Bmax) = 7 pmol/mg protein] receptors for ET-1 were present in myometrial membranes of estrous rabbits. During pregnancy the concentration of ET-1 receptors progressively decreased (Bmax on day 29 = 4 pmol/mg protein), rising again at the time of spontaneous delivery. Conversely, ET-1 receptors in the aorta did not change during pregnancy and parturition. The presence of giant cells immunopositive for ET-1 in close proximity to the myometrial cells, together with the evidence of an increase in ET-1 receptors in myometrium of parturient rabbits, suggests a paracrine role for endometrial ET-1 during delivery. Because we previously demonstrated that oxytocin releases ET-1 from endometrial cells in primary culture and that endometrial and myometrial oxytocin receptors abruptly rise at the time of parturition, we propose that ET-1 might participate in the complex cell-to-cell interactions that occur during labor.

Animals↗
PubMed1992

Spermatic and peripheral venous plasma concentrations of immunoreactive inhibin in prepubertal boys with undescended testis and in pubertal boys with varicocele.

To obtain more information about testicular inhibin secretion in the prepubertal and pubertal human male, we measured the concentrations of immunoreactive inhibin with a heterologous radioimmunoassay in the spermatic and peripheral venous plasma of 5 prepubertal boys with unilateral undescended testis (Group I, P1), 3 prepubertal boys with inguinal hernia (Group II, P1), and 12 pubertal boys with left idiopathic varicocele. The latter subjects were divided, according to the degree of their pubertal development, in early pubertal (Group III, N = 5, P2) and mid-pubertal groups (Group IV, N = 7, P3-4). In Group I, the mean (+/- SD) spermatic venous concentrations of inhibin (289.4 +/- 120.4 ml eq/l) were significantly higher than the corresponding mean peripheral venous concentrations (162.6 +/- 47.2; p < 0.02) suggesting active testicular secretion of inhibin. In Group II, the spermatic-peripheral inhibin gradient was not significant. In pubertal boys with idiopathic varicocele, the mean concentrations of spermatic inhibin were 1076.6 +/- 532.0 and 1023.4 +/- 274.5 in Groups III and IV, respectively. These levels were about five times higher than the corresponding peripheral concentrations (204.8 +/- 41.9 and 238.9 +/- 38.9; p < 0.005 and p < 0.001, respectively). When the data of all the boys were considered together the spermatic venous concentrations of inhibin were significantly correlated with those of peripheral venous FSH (r = 0.4749, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗
PubMed1991

Natriuretic peptide receptors regulate endothelin synthesis and release from parathyroid cells.

Cloned rat parathyroid cells (PTr cell line) that produce parathyroid hormone-related peptide plus endothelin 1 and primary cultures of human parathyroid cells were tested for growth and differentiation responses to atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP). High- and low-affinity binding sites for ANP were found on PTr cells; BNP appeared to bind to the same receptors with similar affinities. Either ANP or BNP stimulated production of cGMP and caused a 30% decrease in Na(+)-K(+)-Cl- cotransport. Each peptide increased synthesis and secretion of endothelin 1 by PTr cells in a dose-dependent fashion, but cell growth was not affected. Human parathyroid cells (normal and pathological) also responded to ANP or BNP with an increase in cGMP production. The finding of receptors for natriuretic hormones on parathyroid cells with consequent effects on release of endothelin 1 might be of relevance in understanding the clinical association between hyperparathyroidism and hypertension.

Animals↗
PubMed1991

Endothelin as an autocrine factor in the regulation of parathyroid cells.

Endothelin, originally purified from porcine aortic endothelial cells, is widely distributed in tissues and is recognized as a product of epithelial cells, glial cells, and neurons in addition to endothelial cells. We found evidence by mRNA content and immunoreactivity that this peptide is synthesized in rat parathyroid epithelial cells (PT-r cells) and bovine parathyroid chief cells. The peptide synthesized by PT-r cells comigrated with synthetic endothelin 1 in reverse-phase HPLC and was diluted out in radioimmunoassay in parallel with the synthetic peptide. Bovine parathyroid endothelial cells (BPE-1 cells) did not express this peptide. Preproendothelin 1 mRNA expression by PT-r cells and endothelin 1 peptide production were regulated by calcium. Shifts in extracellular calcium either from high to low concentrations or vice versa elicited similar evanescent increases in expression of mRNA with a peak at 1 h. Synthesis of the peptide seems to be controlled by mRNA expression, and peptide in the medium appears to be continuously degraded or taken up by cells because its concentration in the medium showed a time course similar to that of mRNA expression. PT-r cells also bear a single class of receptors highly specific for endothelin 1, suggesting an autocrine regulation by endothelin 1 of the parathyroid. The facile regulation of endothelin concentrations in the medium by shifts in extracellular calcium concentration and possible autocrine regulation by endothelin 1 suggest that this peptide may mediate, at least in part, effects of calcium on the parathyroid system.

Animals↗
PubMed1991

Steroid modulation of oxytocin/vasopressin receptors in the uterus.

Oxytocin (OT) and V1 vasopressin (VP) receptors are present simultaneously in several tissues, including the uterus. In myometrium these receptors mediate contractility, while in endometrium they mediate the release of other uterotonic substances as endothelin (ET). In rabbit myometrium, estrogens increase, while progesterone blunts neurohypophysial hormone receptors. However, the action of sex steroids on OT and V1 VP receptors differs in terms of the ED50 and maximal effect. Therefore, at parturition, only OT receptors show a dramatic rise, while V1 VP receptors do not change, suggesting a major role for OT in labor. ET is a potent stimulator of uterine activity acting through specific receptors present on myometrial cells. These receptors as well as the endometrial localization of ET are modulated by sex steroids, indicating that ET might represent a paracrine regulator of uterine activity. In humans, OT but not V1 VP receptors increase as pregnancy progresses, confirming the primary relevance of OT in timing delivery.

Animals↗
PubMed1991

A biomechanical evaluation of a cannulated compressive screw for use in fractures of the scaphoid.

The compressive force generated by a 3.5 mm ASIF cannulated cancellous screw with a 5 mm head was compared with that generated by a standard 3.5 mm ASIF screw (6 mm head), a 2.7 mm ASIF screw (5 mm head), and a Herbert screw. The screws were evaluated in the laboratory with the use of a custom-designed load washer (transducer) to the maximum compressive force generated by each screw until failure, either by thread stripping or by head migration into the specimen. Testing was done on paired cadaver scaphoids. To minimize the variability that occurs with human bone, and because of the cost and difficulty of obtaining human tissue specimens, a study was also done on polyurethane foam simulated bones. The 3.5 cannulated screw generated greater compressive forces than the Herbert screw but less compression than the 2.7 mm and 3.5 mm ASIF cortical screws. The 3.5 mm cannulated screw offers more rigid internal fixation for scaphoid fractures than the Herbert screw and gives the added advantage of placement over a guide wire.

Biomechanical Phenomena↗
PubMed1991

LDH and LDH-X activity in sperm from normospermic and oligozoospermic men.

Total lactate dehydrogenase (LDH) activity was measured in washed sperm from 74 oligozoospermic patients and 148 normospermic men. In the same samples, an additional iso-enzyme specific for germ cells, LDH-X was also measured. Both enzymatic activities were significantly higher in sperm from oligozoospermic patients than in those from normospermic men (P less than 0.001). In oligozoospermic patients, sperm LDH and LDH-X activities were related inversely to sperm concentration (r = -0.61 and r = -0.53, respectively). In normospermic men, this inverse relationship was less evident (r = -0.28 and r = -0.25, respectively). After separation of sperm on a discontinuous gradient of Percoll, LDH and LDH-X activities were increased in the non-migrated-sperm fractions and reduced in the migrated-sperm fractions in both groups of subjects when compared to the initial semen specimens. In addition, migrated sperm from oligozoospermic and normospermic men did not differ in their LDH and LDH-X activities. Although the possible contribution of immature germ-cells to these findings remains to be defined, the data suggest clearly that high levels of intracellular LDH and LDH-X are a biochemical feature of sperm from oligozoospermic subjects.

Centrifugation, Density Gradient↗
PubMed1990

[The Ehlers-Danlos syndrome and its surgical implications].

The Ehlers-Danlos syndrome, type IV, i.e. the arterial and ecchymotic variety, has a severe prognosis and may represent a threat to life in that it predisposes to spontaneous rupture of the intestine and large vessels. The authors report a case in which spontaneous rupture of the colon and thoracic aorta occurred in short succession. Description of the syndrome and its many variants is preceded by a brief explanatory note on the formation and composition of collagen and its different types. The authors stress the important surgical implications of the syndrome the special risks deriving from elective surgery. Serious consequences may also derive from pregnancy trauma and from acts such as arteriography, endotracheal intubation, endoscopy and other invasive procedures. After examining the cases reported in literature (Sacks, Barabas, Beighton Sykes), they point out that, contrary to what is generally believed, the syndrome is not rare and cases, sporadic or familial, of recurrent episodes of spontaneous rupture of the intestine and large vessels or peripheral arteries are frequent. The authors come to the following conclusions: rupture of the aorta may occur spontaneously or after minimal trauma in subjects with EDS type IV and the cases complicated by spontaneous colon perforation would take advantage from definitive colostomy, considering the high incidence of recurrent perforations when intestinal continuity is restored.

Adolescent↗
PubMed1990

Age-associated decline of hepatic handling of cholephilic anions in humans is reverted by S-adenosylmethionine (SAMe).

Decreased fluidity of hepatocyte plasma membrane may contribute to the age-associated changes of liver function. This study aimed at investigating whether the hepatic clearance of organic anions declines with age and whether S-adenosylmethionine (SAMe), a substance proven to be effective in reversing the age-related decrease of membrane fluidity, might influence this process. Nicotinic acid (NA) half-life and serum bilirubin pharmacokinetics after NA load (5.9 mumol/kg body weight i.v.) were studied in 10 healthy young males (YM) aged 14-28 years and in 10 healthy elderly males (EM) aged 65-81 years, before and after SAMe administration (800 mg/day intravenously for 10 days). At baseline, EM showed serum total bilirubin (STB) levels significantly higher than YM. Similarly, the bilirubinaemic mean curves, STB peak and STB time curve concentration after NA load, expressed as area under the curve (AUC), were significantly higher in EM than in YM (p less than 0.01). NA half-life was also significantly prolonged in the aged group (p less than 0.001). SAMe treatment was followed by a significant decrease of basal STB, STB peak and AUC of STB after NA load in EM (p less than 0.01 vs pre-treatment values) while NA half-life was significantly shortened in both groups (p less than 0.001). As NA and bilirubin share a common carrier protein for hepatic uptake, bilitranslocase, the changes observed in EM may be attributed to the reduced lateral mobility of hepatocyte plasma membrane proteins occurring with age. SAMe, by improving membrane fluidity, may increase the diffusion coefficient of bilitranslocase restoring the hepatic handling of organic anions.

Adolescent↗
PubMed1990

Dissociation between vascular and metabolic effects of nicotinic acid in Gilbert's syndrome.

The relationship between the vasodilating and the hyperbilirubinaemic effect of low and high doses (50 and 300 mg i.v.) of nicotinic acid was studied in baseline conditions and after indomethacin pretreatment in healthy controls and patients with Gilbert's syndrome (a condition characterized by fluctuating, nonhaemolytic unconjugated hyperbilirubinaemia). The hyperbilirubinaemic effect of nicotinic acid was confirmed to be more pronounced in Gilbert's syndrome patients than in controls. The magnitude of hyperbilirubinaemia in the two groups was not dependent on the dose of nicotinic acid or indomethacin pretreatment. A dose-dependent vasodilation which was inhibited by indomethacin could be demonstrated in both controls and Gilbert's syndrome subjects. Vasodilating properties of nicotinic acid were therefore found to be dissociated from the effect on bilirubin.

Adolescent↗
PubMed1990

Neurohypophyseal hormone regulation of endothelin secretion from rabbit endometrial cells in primary culture.

Immunoreactive endothelin-1 (IR-ET-1) was detected in the cultured medium from endometrial but not myometrial cells of rabbits in primary culture using a specific radioimmuno assay (RIA). Similar results were obtained with a radioreceptor assay using myometrial membranes. In a reverse-phase HPLC synthetic ET-1 and IR-ET-1 of the extract medium from endometrial cells revealed essentially the same elution profiles, as determined by RIA. Two selective agonists of oxytocin (OT) or V1 vasopressin (VP) receptors produced, respectively, a 6- and 2-fold increase of IR-ET-1 release from endometrial cells. These effects were completely reversed by the addition of two specific antagonists of OT and V1 VP receptors. Our results indicate that ET-1 is produced and released in the culture medium of rabbit endometrial cells in primary culture. The release of ET-1 is under receptor-specific control by neurohypophyseal hormones.

Animals↗
PubMed1989

Identification of a common domain in calmodulin-activated eukaryotic and bacterial adenylate cyclases.

Bordetella pertussis and Bacillus anthracis, two taxonomically distinct bacteria, secrete adenylate cyclase toxins that are activated by the eukaryotic protein calmodulin. The two enzymes contain a well-conserved stretch of 24 amino acid residues [Escuyer et al. (1988) Gene 71, 293-298]. Antibodies have been obtained against two synthetic heptadecapeptides, covering part of the conserved sequences. The anti-peptide antibodies specifically reacted in Western blots with the rat brain adenylate cyclase as well as with the two bacterial enzymes. Anti-rat brain adenylate cyclase serum contained antibodies that were retained by the immobilized peptides, and the affinity-purified antibodies yielded the same recognition pattern of the eukaryotic enzyme as did the unfractionated serum. These results indicate that the eukaryotic adenylate cyclase contains an epitope closely related to that specified by the conserved bacterial sequence. The synthetic peptides and the bacterial adenylate cyclases appeared to compete for ATP (KD of the ATP-peptide complex ca. 0.2 mM), suggesting that the conserved sequence may be part of the substrate binding site in these two enzymes.

Adenylyl Cyclases↗
PubMed1989

Sertoli cell proteins in the human seminiferous tubule.

It is well known that rat Sertoli cells in culture secrete both testis-specific proteins, such as inhibin and androgen binding protein (ABP), and proteins which are very similar, if not identical, to serum proteins, such as transferrin (TF), ceruloplasmin, and IGF-I. It is also well known that very few data have been reported about the secretory activity and the hormonal regulation of the Sertoli cell in man, mainly because of the difficulties associated with the isolation of pure cell populations from human tissue. Using histoimmunochemical techniques we tried to localize, with specific antisera, Sertoli cell proteins and, when possible, their receptors in the human testis. The results obtained with our Light Microscopy studies suggest that: (1) human Sertoli cells produce and/or store transferrin (TF), IGF-I, an albumin-like protein and ABP; (2) TF receptors are localized in spermatocytes and early spermatids and are absent in spermatogonia, in the cytoplasm of Sertoli cells and in differentiated spermatids; (3) IGF-I type I receptors are localized in the same germ cells and in the cytoplasm of Sertoli cells. The results obtained with our Electron Microscopy studies suggest that TF and IGF-I are internalized through a receptor mediated endocytosis mechanism both in Sertoli cells (basal compartment) and in germ cells (spermatocytes and early spermatids).

Androgen-Binding Protein↗
PubMed1988

Insulin-like growth factor-I (IGF-I) and IGF-I receptor in human testis: an immunohistochemical study.

Using specific monoclonal antibodies, the authors studied the distribution of insulin growth factor-I (IGF-I) and its receptor in normal human testis by immunostaining techniques. IGF-I was preferentially localized in Sertoli cells. Less evident positivity was found in primary spermatocytes. In the interstitium some Leydig cells were positive for IGF-I. The major positivity for the IGF-I receptor was found in secondary spermatocytes and early spermatids, whereas Sertoli cells were less positive. An intense positivity was found in some Leydig cells.

Antibodies, Monoclonal↗