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Biomedical subjects

J Martial

Publications and source records attributed to J Martial.

At least 37 records · Page 2Linked to original sources
PubMed1993

The transcriptional regulation of the growth hormone gene is conserved in vertebrate evolution.

Growth hormone (GH) gene expression in mammals is regulated by the interaction of the transcription factor Pit-1 with two binding sites within the proximal promoter. Four sequences, homologous to the mammalian Pit-1 motif occur in the rainbow trout (Oncorhynchus mykiss) GHII (rtGH) gene promoter, two of which partly overlap. The three regions containing these putative Pit-1 binding sequences were protected from deoxyribonuclease I digestion by nuclear extracts of GC cells, a rat pituitary tumor cell line producing Pit-1. In gel shift assays, nuclear proteins from GC cells and from trout pituitaries were found to interact specifically with one of these protected sites. Transfection experiments showed that the rtGH promoter is transcriptionally active in GC cells, the response being strongly enhanced in the presence of a cAMP analogue. The results demonstrate that rat Pit-1 binds to and activates the rtGH promoter, indicating that the basic mechanisms regulating GH gene transcription have been conserved between fish and mammals.

Animals↗
PubMed1993

Effects of prolactin on alpha and beta chloride cells in the gill epithelium of the saltwater adapted tilapia "Oreochromis niloticus".

Tilapia (Oreochromis niloticus), 21 g average body weight, were divided into two groups. A group was maintained in fresh water, whereas another group was adapted for 2 weeks to 20% salt water. Among the latter, fishes were injected every 2 days for a week with tilapia prolactin (ti-PRL I). Gills were prepared for electron microscopy in order to determine the types and surface areas of chloride cells in each experimental condition. Two types of chloride cells, the alpha and beta cells were easily distinguished on the basis of their location and ultrastructural features in the gills of freshwater fishes, while only one type of cell, the saltwater alpha cells presumably derived from the transformation of the freshwater alpha cells, were encountered in saltwater adapted animals. After PRL injection of saltwater adapted fishes, small chloride cells, which displayed ultrastructural features similar to those of beta cells in freshwater tilapia, reappeared in interlamellar regions of the gills. In the same experimental conditions, the voluminous saltwater alpha cells showed a tendency to resume ultrastructural features more characteristic of the freshwater alpha cells from which they were derived. These observations tend to indicate that prolactin behaves as a "freshwater adapting hormone" and that beta cells are specifically involved in fish adaptation to freshwater living conditions.

Adaptation, Physiological↗
PubMed1993

Membrane translocation and relationship with MHC class I of a human thymic neurophysin-like protein.

Thymic epithelial and nurse cells (TEC/TNC) synthesize an oxytocin (OT)-like peptide in association with a neurophysin (NP)-related protein in a way similar to in the hypothalamo-neurohypophysial (NHP) system. The central T-cell tolerance of the NHP neuroendocrine functions have been proposed to be mediated through these thymic NHP-related peptides due to their close homology with the NHP neurohormones OT and vasopressin (VP). In order to investigate their putative presentation by proteins of the major histocompatibility complex (MHC), human thymic membranes were purified and passed through an immunoaffinity column using mAb B9.12 directed to the monomorphic determinant of human MHC class I proteins. This methodology provided the following observations: (1) a NP-like protein is translocated in human thymic membranes and is retained by B9.12 on the column; (2) the MW of this NP-like material (50-55 kD) is quite different from the MW of hypothalamic NP proteins (10 kD), and (3) this thymic NP-like protein could be identified on Western blots with mAb B9.12. The precise extent of this relationship between the thymic NP-like protein and the Ig/MHC superfamily is actually investigated through the characterization of the genetic mechanisms responsible for the thymic expression of NHP-related peptides. Given the physiological importance of OT and of its binding to NP for transport along the axonal processes of the NHP tract, we postulate that, somewhat analogously, the thymic NP-/MHC class I-related protein could be involved in the presentation of the OT-like peptide to immature T-cells.

Amino Acid Sequence↗
PubMed1991

Characterization of 5-HT1D receptor binding sites in post-mortem human brain cortex.

The present study provides further evidence for the presence of serotonin1D (5-HT1D) receptors in post-mortem human brain. Receptor binding parameters in temporal cortex homogenates were assessed using [3H]5-HT in the presence of 100 nM 8-OH-DPAT, 1 microM propranolol and 1 microM mesulergine to prevent labelling of the 5-HT1A, 5-HT1B and 5-HT1C sites, respectively. Under these conditions, [3H]5-HT apparently bound to a class of high affinity (Kd = 5.0 +/- 1.0 nM) low capacity (Bmax = 96 +/- 23 fmol/mg protein) sites. In competition experiments, 5-HT and 5-carboxyamidotryptamine (5-CT), as well as ergotamine, lysergic acid, sumatriptan and RU-24969 exhibited high affinity for these sites. This pharmacological profile is concordant with the ligand selectivity pattern reported for 5-HT1D receptors in other species and thus provides further evidence for its existence in human temporal cortex. In addition, the competition profile of some ligands, particularly of unlabelled 5-HT, 5-CT and ergotamine, revealed the existence of a lower affinity binding site. The latter suggests receptor heterogeneity or the presence of a lower affinity state of 5-HT1D receptors.

Adolescent↗
PubMed1990

Growth hormone and somatostatin gene expression in pituitary adenomas with active acromegaly and minimal plasma growth hormone elevation.

Some patients with active acromegaly have elevated plasma IGF-I concentrations with only minimal elevation of plasma GH. We compared adenomatous GH and SRIH expression in 3 such patients (patients No. 1, 2 and 3; basal plasma GH level less than 4 micrograms/l) and in 3 acromegalic patients with high basal plasma GH level (patients No. 4, 5 and 6; 51.7 +/- 16.1 micrograms/l, mean +/- SEM). By immunocytochemistry, all the tumours proved to be somatotropic adenomas. At the ultrastructural level, signs of low secretory activity were observed in adenomas from patients No. 2 and 3. Perifused adenoma cells of patients No. 1, 2 and 3 released very little GH compared with those of patients No. 4, 5 and 6 (1 +/- 0.37 vs 51.5 +/- 34.1 micrograms x (10(-6) cells) x min-1, p less than 0.001). Adenoma SRIH content was 65.7 and 30.6 pg/mg proteins in patients No. 1 and 2, whereas it was undetectable in the others (patients No. 4, 5 and 6). Northern blot analysis showed that the GH gene was poorly expressed in the adenomas from patients No. 1, 2 and 3 compared with the adenomas from patients No. 4, 5 and 6. SRIH mRNA was detected in all 6 adenomas. However, the signal was more intense in the adenomas from patients No. 1, 2 and 3 than in those from patients No. 4, 5 and 6.(ABSTRACT TRUNCATED AT 250 WORDS)

Acromegaly↗
PubMed1989

Apparent absence of serotonin1B receptors in biopsied and post-mortem human brain.

In order to determine if post-mortem delays could account for the apparent absence of 5-HT1B sites reported in human brain, 5-HT1B binding parameters were determined in fresh and fresh-frozen cortical biopsies and compared to results obtained in post-mortem tissues. Binding parameters and in vitro receptor autoradiography were performed by using two different ligands which have been shown to label 5-HT1B sites, namely [3H]5-HT, in presence of 100 nM 8-OH-DPAT, and [125I]cyanopindolol, in presence of 10 microM (-)isoproterenol. No specific binding was detected with [125I] cyanopindolol in either fresh cortical biopsies or post-mortem tissues, suggesting that the apparent absence of 5-HT1B receptor sites reported earlier was not related to long post-mortem delays. Some specific labelling was seen with [3H]5-HT, in presence of 8-OH-DPAT. This binding, which is unlikely to be to the 5-HT1B type, could represent labelling to the 5-HT1D or 5-HT1E sites.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1988

Pairing properties of bromouracil and repair of bromouracil-containing DNA. Possible utilization of bromodeoxyuridine triphosphate for site-directed mutagenesis.

5-Bromo-2'-deoxyuridine triphosphate (Br-dUTP) and dTTP are used interchangeably for DNA synthesis in vitro by the Klenow fragment of Escherichia coli DNA polymerase I. When DNA containing Br-dUMP instead of dTMP at a few preselected sites is transfected into competent bacteria, no mutation occurs, indicating that in vivo E. coli DNA polymerase always places a dAMP residue in front of any unrepaired Br-dUMP residue. On the other hand, in vitro Br-dUTP can also replace dCTP, but only with difficulty: when dCTP is absent, Br-dUMP can be forced in front of a dGMP residue, but the Klenow polymerase pauses before and after addition of Br-dUMP. Transfection into E. coli of the substituted DNA leads to the expected G----A transitions. These mutations can easily be targeted by using a suitable primer and the correctly chosen mix of deoxynucleoside triphosphates containing Br-dUTP. When Br-dUMP has been placed in front of a dGMP residue, the mutation yield is not 100%, showing a partial repair of the transfected DNA before it is replicated. Advantage can be taken of this partial repair to prepare a set of different mutations within a target region in a single experiment.

Base Composition↗
PubMed1987

Human brain receptor autoradiography using whole hemisphere sections: a general method that minimizes tissue artefacts.

A general method for the preparation of high-quality, mostly ice-crystal-artefact-free whole human brain hemisphere sections is described. Upon receipt, hemispheres are divided; one is then fixed in buffered 10% formalin for neuropathological analysis while the other is cut in 8-10-mm-thick coronal slices that are then rapidly frozen in 2-methylbutane at -40 degrees C (10-15 sec) before being placed in the brain bank at -80 degrees C. Such rapid freezing markedly decreases the formation of ice-crystal artefacts. Whole-hemisphere 20-micron thick sections are then cut and mounted onto lantern-type gelatin-coated slides. These sections are subsequently used for both qualitative and quantitative in vitro receptor autoradiography. Examples of data obtained are given by using various radioligands labelling "classical" neutrotransmitter, neuropeptide, enzyme, and ion channel receptor binding sites. This method should be useful for the obtention of various receptor maps in human brain. Such information could be most useful for in vivo receptor visualization studies using positron emission tomography (PET) scanning. It could also indicate if a given receptor population is specifically and selectively altered in certain brain diseases, eventually leading to the development of new therapeutic approaches.

Adult↗
PubMed1987

Expression of the growth hormone variant gene in human placenta.

Besides the hGH-N gene, which codes for the pituitary 22 and 20K GH variants, the human genome contains a second GH gene, namely the GH-V, which has been thought to be silent. We recently discovered a placental variant of human growth hormone (hPGH), which appears in maternal serum at mid-pregnancy and which rises in concentration thereafter to term. As hPGH and GH-V proteins display very similar characteristics, including a high affinity for hepatic GH receptors, they could be identical. To verify this hypothesis, we sought hGH-V mRNA in placenta. Hybridization experiments were performed between dot-blotted mRNA originating either from placenta or from one pituitary hGH secreting adenoma and synthetic polynucleotide probes corresponding to specific portions of the hGH-V or hGH-N gene sequences. The results indicate that the V gene is indeed expressed in the placenta and, at a very low level, in the pituitary adenoma. Therefore hPGH is most likely the expression product of the hGH-V gene.

Adenoma↗
PubMed1985

Sequestration of 3H-vitamin D3 by the fetal and neonatal rat liver.

19- to 21-day-old fetuses as well as 3-, 7-, 14-, and 22-day-old pups were used to evaluate the sequestration of a tracer dose of 3H-D3 by the fetal and neonatal rat liver. In fetuses, 3H-D3 was injected directly into the umbilical vein; in neonates, 3H-D3 was injected into the portal vein. In both cases, 14C-sucrose was used as extracellular marker. 3H-D3 sequestration was evaluated at 11.3 +/- 1.1% (means +/- SEM) in fetuses, 3.3 +/- 0.6, 4.9 +/- 0.6, 6.3 +/- 0.5 0.5 and 42.0 +/- 1.9% of the dose administered in 3-, 7-, 14-, and 22-day-old rats, respectively. These results clearly show that, in the rat, the fetal liver can take up 3H-D3. Moreover, when compared to the values obtained in the late fetal period, 3H-D3 uptake decreased significantly during the first 2 weeks post partum (p less than 0.0001) and then dramatically increased (6-fold, p less than 0.0001) between days 14 and 22 post partum, when it reached an uptake capacity similar to that previously observed in adult rats.

Age Factors↗
PubMed1981

Regulation of calcium-binding protein messenger RNA by 1,25-dihydroxycholecalciferol.

1,25-Dihydroxycholecalciferol modulates the synthesis of the intestinal calcium-binding protein. To determine if this effect is due to an increase in calcium-binding protein mRNA activity, we measured total cytoplasmic protein-specific mRNA activity from chick intestine in an in vitro wheat germ translation system. 1,25-Dihydroxycholecalciferol enhances calcium-binding protein specific mRNA activity. The sterol does not induce a general increase in mRNA synthesis since the concentration of only a few proteins increased. Thus 1,25-dihydroxycholecalciferol regulates only a subset of genes and specifically affects a transcriptional process involving calcium-binding protein synthesis.

Animals↗
PubMed1980

The pH dependence of rat liver RNA polymerases I and II.

The effect of pH on the stability and activity of rat liver RNA polymerases I (A) and II (B) has been studied. Both enzymes are irreversibly inactivated in buffer solutions below pH 5.0. Km values of the two enzymes are constant between pH 6.5 and 8.7 but a two -to three- fold increase is observed between pH 8.7 and 9.7. The Vmax versus pH profiles are bell-shaped curves indicating the participation of two ionizing groups with apparent pKa values of 6.5 and 9.8 for enzyme I and 6.7 and 9.9 for enzyme II. Both enzymes are inactivated by photooxidation in the presence of Rose Bengal. It is suggested that the above pKa corresponds to the imidazole of a histidine residue and a amino group of a lysine residue.

Animals↗