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Biomedical subjects

A Gerard

Publications and source records attributed to A Gerard.

At least 37 records · Page 2Linked to original sources
PubMed1995

Endocytosis of androgen-binding protein (ABP) by spermatogenic cells.

To test whether Sertoli cell-secreted ABP could serve as steroid carrier to the germ cell (GC) lineage, radiolabeled ABP and SHBG and gold SHBG were used for binding studies and for internalization studies based on transmission electron microscope analyses and autoradiography of the radiolabeled samples. The data clearly showed that: (1) rat and human germ cells possess a single class of binding sites for rat ABP and human SHBG respectively (Kd of 0.78 and 0.56 nM); (2) 1.7 x 10(10) and 2.7 x 10(10) sites/mg protein was found in the corresponding plasma membrane preparations; (3) the receptor peak was eluted in the same position as dextran blue: 2000 kDa (M(r) = 2 x 106) for labeled rat ABP; (4) in the whole GC lineage, the labeled ligand was internalized through an endocytic pathway involving clathrin coated structures and the distribution was similar throughout the maturation step, however striking differences in the internalization rate were revealed with regard to the maturation step; and (5) this internalization occurred even in ligated seminiferous tubules, via the Sertoli cells cytoplasm. When isolated rat GC were incubated in the presence of ABP, a dose dependent increase in labeled secreted protein was observed for spermatocytes (50-250%) whereas ABP had no effect on spermatids. Addition of steroids and ABP caused a 200 and 50% increase in labeled secreted proteins for spermatocytes and spermatids respectively. 2-D SDS-PAGE analysis revealed that ABP alone increased the secretion of specific spermatocyte proteins whereas steroids in the presence of ABP resulted in the synthesis of new spermatocyte secreted proteins. Taken together these results strongly suggest that ABP may be required for spermatogenesis either as a steroid transmembrane carrier or on its own.

Androgen-Binding Protein↗
PubMed1995

Endogenous glutamate involvement in pulsatile secretion of gonadotropin-releasing hormone: evidence from effect of glutamine and developmental changes.

The secretion of GnRH can be stimulated by glutamate (GLU) and GLU agonists, whereas GLU receptor antagonists inhibit GnRH. Using 6-diazo-5-oxo-L-norleucine (DON), an inhibitor of glutaminase, we aimed to study the involvement of endogenous GLU in GnRH secretion through the effects of impaired GLU biosynthesis from its precursor glutamine (GLN). GnRH secretion by hypothalamic explants of male rats, aged 15 and 50 days, was compared, because the frequency of spontaneous GnRH secretory pulses showed a 2-fold increase between those two ages. Using explants of 50-day-old rats, GLN elicited GnRH secretion in a similar dose-related manner as GLU. DON prevented GLN-evoked secretion of GnRH, whereas the effect of GLU was not altered. DON also markedly inhibited spontaneous pulsatile secretion of GnRH and the secretory response to veratridine, a Na+ channel opener. The inhibitory effect of DON on veratridine-evoked secretion of GnRH was directly related to the duration of exposure to DON and the frequency of GnRH secretory episodes. Using explants of 15-day-old rats, GLN could elicit GnRH release, although this response was lower than GLU-evoked secretion of GnRH. The DON concentrations required for inhibition of veratridine-evoked secretion of GnRH were lower at 15 days than at 50 days. These data indicate that 1) GLU biosynthesis from GLN is a prerequisite to the physiological mechanism of pulsatile GnRH secretion; and 2) inhibition of veratridine- or GLN-induced secretion of GnRH requires higher DON concentrations after the onset of puberty than before. This suggests that glutaminase, the enzyme controlling GLU biosynthesis from GLN, shows increased activity after the onset of puberty when the frequency of pulsatile GnRH secretion is increased as well.

Aging↗
PubMed1995

Survival of patients with radiation enteritis of the small and the large intestine.

We reviewed the clinical and survival data of 108 consecutive patients who presented with radiation enteritis between 1965 and 1981. One hundred and two (94%) had been irradiated for carcinoma of the cervix uteri. The median follow-up was of 11 years. The median time of occurrence of severe radiation-induced lesions (obstruction, perforation) after radiotherapy was of 18 months, against 10.5 months for mild symptoms (e.g., tenesmus) and 9 months for rectal bleeding (p < 0.001). Cox survival analysis taking into account the stage of the cancer and the age at diagnosis showed that rectal bleeding is a factor with a prognosis significantly poorer than the mild symptoms (p = 0.05), equivalent to that of the severe complications. We conclude that in the evaluation of patients who underwent radiotherapy for abdominal or pelvic tumours, rectal bleeding should be considered as a sign of serious radiation-induced complication.

Abdominal Neoplasms↗
PubMed1994

Spermatogenic cells do internalize Sertoli androgen-binding protein: a transmission electron microscopy autoradiographic study in the rat.

Specific binding sites for androgen-binding protein (ABP) have been demonstrated recently to be present on germ cells from the rat and on membrane-enriched fractions from rat germ cells. The present study was undertaken to test if such receptors could lead to the activation of a specific internalization pathway as in other cells. Isolated rat germ cells and in situ rat germ cells, maintained within the seminiferous epithelium with either an intact or a bypassed blood testis barrier, were exposed to culture medium containing 12,000 cpm/ml [3H] delta 6-testosterone (2.5 pg) photoaffinity-labeled ABP, purified from rat testis. The follow-up of labeled ABP/germ cell interactions was based on qualitative and quantitative transmission electron microscopy autohistoradiography. Attention was focused on adluminal germ cells from pachytene spermatocytes to mature spermatids, which are normally present above the Sertoli cell tight junctions. Our observations revealed the presence in rat germ cells of structures related to specific endocytosis, namely coated pits and vesicles which stained positively with anticlathrin antibodies. When exposed to the [3H] delta 6-testosterone-ABP complex, adluminal germ cells showed marked labeling of these endocytic organelles. Preincubation either with excess unlabeled ABP or pretreatment by EDTA reduced the labeling significantly. Once internalized, ABP was found to be confined to the endocytic and nuclear compartments. The nuclear labeling was high in primary spermatocytes and round spermatids but was absent in elongated spermatids with condensed chromatin, in which transcriptional activity had almost stopped. In contrast, at later steps of spermiogenesis, the cytoplasm became heavily labeled, especially in the postnuclear part of elongated spermatids and in residual bodies about to be phagocytozed by Sertoli cells. Experiments using ligated seminiferous tubules, with an intact blood-testis barrier, clearly showed that ABP was captured at the basal part of Sertoli cells, transported up to the adluminal compartment and delivered to germ cells which finally internalized the protein. Experiments using largely opened seminiferous tubules allowing ABP to bypass the blood-testis barrier led to a delay in adluminal germ cell labeling compared with that in isolated germ cells. In all experiments in which germ cells were incubated in teh presence of Sertoli cells, the labeling observed at the surface of the germ cell line, especially over coated pits, was mostly found facing thin Sertoli cell processes, suggesting the possible existence of a specific mechanism for the presentation of ABP by the Sertoli cells to adluminal germ cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Androgen-Binding Protein↗
PubMed1994

Gonadotropin releasing hormone inhibitory autofeedback by subproducts antagonist at N-methyl-D-aspartate receptors: a model of autocrine regulation of peptide secretion.

The secretion of Gonadotropin-releasing Hormone (GnRH) involves activation of N-methyl-D-aspartate (NMDA) receptors. Here, we show that pulsatile GnRH secretion from hypothalamic explants is suppressed by 1-5GnRH, an endogenous breakdown product of GnRH, while 2-10GnRH has no effect. GnRH secretion evoked by NMDA is selectively inhibited by 1-5GnRH and this effect is similar to that of AP-5, a competitive antagonist at NMDA receptors. In addition, 1-5GnRH accounts for a dose-related inhibition of tritiated glutamate binding to hypothalamic membrane preparations. Using GnRH secretion as a model of NMDA-receptor controlled system, the effect of different peptides has been studied. Growth Hormone Releasing Factor (GRF), Insulin-like Growth Factor-I (IGF-I) and Proinsulin result in inhibition of GnRH secretion. Bioactive subproducts of those peptides (1-29GRF, 4-701GF-I and insulin) do not show any effect, suggesting that their classical receptors are not involved. In contrast, GnRH secretion is inhibited by other subproducts (1-37GRF, 1-31GF-I and C-peptide) all terminating in a glutamate residue. These subproducts selectively suppress the NMDA-evoked secretion of GnRH. Protease inhibitors prevent the inhibitory effects of IGF-I on GnRH secretion. This, breakdown products of different peptide hormones are possible endogenous antagonists at NMDA receptors. This effect could account for an autocrine or paracrine limitation of NMDA-receptor-mediated secretion of peptides.

Animals↗
PubMed1994

A surgical model for studying biliary excretion of drugs in the awake micropig yucatan.

The pig is probably one of the best mammalian models for studying the digestive functions and the biliary excretion of drugs because of the tight similarities with humans. Insertion of a T drain in the common bile duct after cholecystectomy in humans became uncommon because of surgical progress. For this reason, we developed a model using a bidirectional cannula to study the biliary secretion in the conscious Micropig Yucatan. We used this breed because of their ready availability and low stable weight. The surgical procedure was a double choledococholedocal fistula with a bypass on the main biliary duct, maintaining the continuous flow of bile into the duodenum through the sphincter of Oddi. This fistula remained effective for at least sixty days with normal hepatic biologic parameters. The development of our model could improve the biliary excretion study of drugs. Comparison of different routes of administration and interaction pharmacokinetic studies could also be studied in the same micropig, eliminating intersubject variations.

Animals↗
PubMed1993

Endocytosis of transcobalamin in male rabbit germ cells: electron microscope radioautography study.

The binding of 125I-iodinated transcobalamin to a suspension of isolated rabbit germ cells was studied by Scatchard plot. The number of binding sites was evaluated to about 1000 per cell, and its association constant (Kass) in order of 14.6 l/nmole. The distribution to structures related to endocytosis was determined by ultrastructural histomorphometric studies. Both coated and uncoated structures were present regardless of maturation stage. The number of coated vesicles was at its highest in the initial maturation steps, whereas the number of uncoated vesicles was highest in the final maturation steps. The endocytosis of 125I-iodinated transcobalamin by the suspension of germ cells was studied by electron microscope radioautography. The tracer was mostly detected over the plasma membrane, coated vesicles and multivesicular bodies of germ cells. The grains were observed mainly over spermatocytes and round spermatids; 31.6% and 32% of these cells, respectively, were labeled. In contrast the tracer was detected in only 8.11% of elongated spermatids. In conclusion, iodinated transcobalamin is internalized in rabbit germ cells by receptor-mediated endocytosis. This phenomenon was predominant in the early stages of germ cell maturation.

Animals↗
PubMed1992

[Antibioprophylaxis in pulmonary surgery: a randomized trial with cefamandole versus placebo].

A prospective double-blind study was designed to assess the efficacy of antibiotic prophylaxis in lung surgery. It included 114 patients undergoing lung surgery for primary or secondary malignant tumours randomly assigned to two groups. Group A patients (n = 59) were given cefamandole intravenously every four hours, three times, starting from induction of anaesthesia. The dose was determined according to the patient's weight: 1.5 g for patients weighing less than 60 kg, 2.5 g for those weighing between 60 and 80 kg, and 3 g for those above 80 kg. Group B patients (n = 55) were given a placebo at the same times. Nineteen other patients were excluded because either the tumour was found to be infected, or the patient had to be mechanically ventilated postoperatively, or an exploratory thoracotomy only was carried out, or they were allergic to beta-lactam antibiotics. The efficacy of antibiotic prophylaxis was assessed by recording the incidence of postoperative infections, the length of the patient's stay in hospital, and the need to use an antibiotic treatment. Patients, their sputum and wound were examined every day, and their temperature recorded. The white blood cell count and chest X-ray was carried out every day for the first week. All the drain and catheter tips were cultured, as well as sputum and blood (every three days). In case of infection, samples were obtained and cultured. Both groups of patients were similar with regard to age, risk factors (smoking habit, diabetes mellitus), and type of surgery (segmentectomy, lobectomy, pneumonectomy). There were 9 postoperative infections in group A, and 22 in group B (p = 0.003).(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1992

Priming doses of atracurium and vecuronium depress swallowing in humans.

The administration of low doses of muscle relaxant may cause peripheral muscular weakness including difficulty in swallowing. In the present study, the effect of priming doses of atracurium and vecuronium on swallowing was studied. Sixty patients undergoing elective surgery under general anesthesia were divided randomly into four groups of 15 patients and received as a priming dose either vecuronium (10 or 15 micrograms/kg) or atracurium (50 or 75 micrograms/kg). Swallowing muscle activity was measured by electromyography using submental surface electrodes. Swallowing was initiated by administration of 0.3 ml distilled water through an oral catheter. Swallowing reflex was determined by measuring the latency time (i.e., time from water administration to start of EMG activity of glossal muscles). Swallowing activity was determined by integration of the EMG of glossal muscles during swallowing. Peripheral muscle strength was determined by hand grip strength. Swallowing reflex activity and peripheral muscle strength were measured before and 3 and 6 min after administration of vecuronium or atracurium. Latency time remained unchanged after any of the priming doses. Integrated EMG decreased significantly (P < .001) 3 and 6 min after all priming doses tested (42-75% of baseline value). Only after atracurium 75 micrograms/kg was the hand grip strength significantly decreased (P < .01). These results suggest that owing to its effect on swallowing, the priming dose should be used with caution.

Adult↗
PubMed1991

Endocytosis of human sex steroid-binding protein in monkey germ cells.

We investigate in this study the hypothesis of human sex steroid-binding protein hSBP internalization into germ cells in a primate model. Human SBP was purified from late-pregnancy serum and labeled either with colloidal gold particles (18 nm) or with [3H]delta 6-testosterone by photoaffinity treatment. The germ cells were isolated from sexually mature monkey testis or caput epididymis (Macaca fascicularis) by mechanical means and cell suspensions (4 x 10(6) per 100 microliters culture medium) were incubated in presence of hSBP-gold complex (60 ng/100 microliters) or hSBP-[3H]delta 6-testosterone complex (66 ng/100 microliters, 20,000 cpm) for 2, 5, 15, 45, and 60 min. The samples were processed for electron microscopy followed by autoradiographic treatment for the radiolabeled samples. Localization of the label occurred over the whole germ cell lineage whichever tracer was used. Spermatogonia, spermatocytes, spermatids, testicular and epididymal spermatozoa exhibited specific binding sites over the plasma membrane associated with clathrin-like coated pits and vesicles. At 34 degrees C, intracellular localization of the labeled ligand was found within coated vesicles, in early and late endosomes. In addition, in early spermatogenic cells, labeled ligand was detected in the nuclei and/or associated with the nuclear envelope whereas in late spermatids and residual bodies, the labeling was accumulated in multivesicular, prelysosomal structures. Quantitative analysis of the "labeled cells/total cells" ratio exhibited a negative correlation to the maturation steps, epididymal spermatozoa being the least labeled. The cellular distribution is similar with one or the other protein in the same spermatogenic cells. Unlabeled hSBP treatment prior to labeled hSBP reduced significantly the internalization. Lowering the temperature to 4 degrees C prevented endocytosis and enhanced membrane binding. EDTA pretreatment strongly decreased hSBP internalization and modified the early endocytic steps, namely, the pinching off of the coated vesicles. It is concluded that monkey germ cells are able to internalize the human sex steroid-binding protein through specific endocytic organelles. This endocytosis leads to the labeling of the nuclei in the early spermatogenic cells and of the multivesicular bodies in the late germ cells. This strongly suggests that steroid-binding proteins may be required for spermatogenesis in acting at the germ cell lineage level either by themselves or by serving as steroid transmembrane carriers.

Animals↗
PubMed1991

Evidence that androgen-binding protein endocytosis in vitro is receptor mediated in principal cells of the rat epididymis.

We have studied the binding of [125I-iodo]androgen-binding protein (ABP) and of [3H]delta 6-testosterone photoaffinity-labelled ABP to receptors in the plasma membrane of rat epididymal cells in three ways: ABP binding to a Triton X-100-solubilized membrane extract, ABP binding to isolated epithelial cells in suspension and autoradiography of segments of dissected epididymides after in-vitro intraluminal injection of labelled ABP. The binding of iodinated ABP to the receptor was similar to that of photoaffinity-labelled ABP in gel filtration. The ABP-receptor complex was eluted from Superose 6 gels as an aggregate, with a molecular mass of 2000 kDa. It was separated into two peaks by sucrose gradient ultracentrifugation, with respective sedimentation coefficients of 18.4 and 9.0 s. The activity of the receptor (ABP-binding capacity/mg protein) was tenfold higher in the caput than in the cauda. The binding of ABP to the receptor was pH dependent, being almost abolished at pH less than 4. The binding at 4 degrees C of photoaffinity-labelled ABP to epithelial cells corresponded to two types of binding sites. The numbers of high-affinity and low-affinity sites per cell were 1600 and 7700 respectively; the association constants of these sites were 67.9 and 2.8 litres/nM respectively. The binding was decreased by treatment of the cells with trypsin or incubation in the presence of EDTA. The binding in vitro of labelled ABP to the epididymis epithelium reached a maximum after about 20 min at 4 degrees C. In the autoradiographic study the tracer was found to be closely associated with coated pits, coated vesicles, endosomes and pale multivesicular bodies. Treatment of rats with cycloheximide significantly reduced the uptake of the tracer. Perfusion in vitro of epididymides with chloroquine produced a fourfold increase of the tracer in endosomes and multivesicular bodies.

Androgen-Binding Protein↗
PubMed1991

Hepatic artery ligation with and without portal infusion of 5-FU. A randomized study in patients with unresectable liver metastases from colorectal carcinoma. The E.O.R.T.C. Gastrointestinal Cancer Cooperative Group (G.I. Group).

The aim of this multicentric prospective randomized clinical trial was to study the efficacy of hepatic artery ligation (HAL) with and without portal infusion (PI) of 5-FU in patients with liver metastasis of colorectal origin. Seventy-four patients were randomized. Sixty-seven were fully evaluable. Thirty-five patients were eligible in the HAL + PI of 5-FU group and 32 in the HAL alone group. The 5-FU infusion had to be discontinued for technical reasons in 13 patients. Complications of HAL were relatively high, including four hepatic failures (WHO grading greater than 2). Side effects of chemotherapy were limited. Five patients out of 30 had a partial response (WHO criteria) and one patient had a complete response in the group treated by HAL and PI of 5-FU. Only one patient had a partial response in the HAL alone group. Median survival for both groups was 12 months. Median time to progression for both groups was 6 months. This study did not show any advantage of delivery using the portal route in addition to hepatic artery ligation in terms of progression nor in survival of patients.

Adenocarcinoma↗