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Biomedical subjects

A Israel

Publications and source records attributed to A Israel.

At least 73 records · Page 4Linked to original sources

Effects of adrenalectomy and aldosterone on the action of centrally administered rat atrial natriuretic peptide-(99-126).

Intracerebroventricular (i.v.t.) administration of rat atrial natriuretic peptide-(99-126) (rANP) increases urinary volume and sodium excretion, but the mechanism is undefined. A diminished mineralocorticoid effect on the kidneys may explain the natriuretic phenomenon. This hypothesis was tested by i.v.t. rANP injection (1.25 micrograms/5 microliters) in conscious, hydrated rats pretreated beforehand with d-aldosterone (20 micrograms/kg, i.p.). Although the absolute amount of sodium excreted was reduced, aldosterone did not affect rANP-induced sodium output at 1 and 3 h. Rats which were sham-operated or bilaterally adrenalectomized (ADX) after four days were pretreated with aldosterone and given an oral water load followed by i.v.t. rANP or saline. In ADX rats natriuresis and diuresis after rANP were still evident. Our results indicate that the natriuretic effect of i.v.t. rANP is unrelated to plasma levels of mineralocorticoids. Likewise, diuresis and natriuresis can occur in the absence of the adrenal glands.

Adrenalectomy↗

Rat atrial natriuretic peptide (99-126) stimulates guanylate cyclase activity in rat subfornical organ and choroid plexus.

The effect of rat atrial natriuretic peptide (99-126) (rANP) on cyclic guanosine 3',5'-monophosphate production was investigated in two brain areas, the subfornical organ and the choroid plexus. rANP activated guanylate cyclase in crude homogenates of rat subfornical organ and choroid plexus in concentration and time dependent fashions. A 2-fold stimulation of the enzyme was obtained with 100 nM rANP and a half-maximal stimulation with a 10 nM dose. Our results further support the hypothesis that cGMP mediates the central action of ANP through the activation of specific receptors in localized target sites such as the subfornical organ and choroid plexus.

Animals↗

Natriuretic and diuretic action of centrally administered rat atrial natriuretic peptide (99-126): possible involvement of aldosterone and the sympathoadrenal system.

Intracerebroventricular (ICV) administration of rat atrial natriuretic peptide (99-126) (rANP) to conscious male hydrated rats resulted in a dose-related increase in urinary volume and sodium excretion over a 6-h period of urine collection. A diminished mineralocorticoid effect on the kidneys may explain the natriuretic phenomenon. This hypothesis was tested by ICV rANP injection (1.25 microgram/5 microL) in conscious hydrated rats pretreated beforehand with d-aldosterone (20 micrograms/kg, ip). Although the absolute amount of sodium excreted was reduced, aldosterone did not affect rANP-induced sodium output at 1 and 3 h. Rats that were sham-operated or bilaterally adrenalectomized after 4 days were pretreated with aldosterone and given an oral water load followed by ICV rANP or saline. The possible participation of the peripheral sympathetic nervous system in the central action of rANP was evaluated in rats pretreated with 6-hydroxydopamine. In sympathectomized and adrenalectomized rats natriuresis and diuresis were still evident after rANP. Our results indicate that the natriuretic effect of ICV rANP is independent of mineralocorticoids. Likewise, diuresis and natriuresis can occur in the absence of the adrenal glands and are independent from the neural tone that the adrenergic system exerts on sodium reabsorption.

Adrenal Glands↗

Isolation of the lux genes from Photobacterium leiognathi and expression in Escherichia coli.

Genes necessary for luminescence (lux genes) in the marine bacterium Photobacterium leiognathi, strain PL721, were isolated and expressed in Escherichia coli. A 15-kb fragment obtained from a partial digestion of PL721 DNA with HindIII was cloned into the plasmid pACYC184, resulting in the hybrid plasmid pSD721. When pSD721 was transformed into E. coli ED8654, the resulting transformants were luminous with no additions to the cells, indicating that it contained the structural genes coding for the alpha and beta subunits of luciferase (luxA and luxB), and for components involved in aldehyde biosynthesis. Hybridization analysis with luxA and luxB 32P probes confirmed the location of these two genes on the 15-kb insert. When pSD721 was transformed into four different strains of E. coli, luminescence expression varied widely in amount and in pattern. In some strains, luminescence developed like an autoinducible system, and at maximum induction was very bright, even with no addition of aldehyde, while in others, luminescence was 100-fold less, and no induction was seen. In no case was luminescence affected by shifts in temperature, osmolarity, or iron concentration. These results indicate that, while the complete lux regulon is apparently contained on the 15-kb cloned fragment, the regulation of the lux regulon in pSD721 is subject to host controls by E. coli, controls which vary widely among different E. coli strains.

Cloning, Molecular↗

High angiotensin converting enzyme (kininase II) activity in the cerebrospinal fluid of spontaneously hypertensive adult rats.

Angiotensin converting enzyme (ACE, Kininase II, E.C. 3.4.15.1) activity was measured in the cerebrospinal fluid of 4- and 16-week-old spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto (WKY) normotensive controls. Adult SHR showed higher cerebrospinal fluid enzyme activity than normotensive age-matched WKY (19.6 +/- 1 and 32.3 +/- 5 nmol/h per ml in WKY and SHR, respectively, P less than 0.025). Conversely, there were no significant differences in enzyme activity in the cerebrospinal fluid of young animals. Our results support the hypothesis of enhanced activity of the central angiotensin system during the established phase of spontaneous hypertension in rats.

Age Factors↗

Increased atrial natriuretic peptide binding sites in the rat subfornical organ after water deprivation.

Binding sites for rat atrial natriuretic peptide (99-126) were localized and quantified in the subfornical organ and choroid plexus of control and water-deprived rats. Brain sections were analyzed after incubation with 125I-rat atrial natriuretic peptide (99-126), autoradiography, computerized microdensitometry and comparison to 125I-standards. Higher numbers of atrial natriuretic peptide binding sites were present in both the subfornical organ and the choroid plexus after 4 days of water deprivation. Our results indicate a central role for atrial natriuretic peptide in the regulation of water balance.

Animals↗

Compensatory increase in adrenomedullary angiotensin-converting enzyme activity (kininase II) after unilateral adrenalectomy.

Angiotensin-converting enzyme (ACE, kininase II, dipeptidyl carboxypeptidase, EC 3.4.15.1) was characterized in the adrenal medulla of male Sprague-Dawley rats. Rat adrenal medulla and lung ACE were similar in their susceptibility to Cl- activation and to the inhibition by EDTA, captopril, bacitracin and thiorphan, suggesting that rat adrenal medulla and lung ACE have similar properties. Changes in right adrenal weight and in adrenomedullary ACE activity 5 and 12 days following left unilateral adrenalectomy (UADX) were examined. Compensatory adrenocortical hypertrophy 12 days following UADX was associated with a significant increase in adrenal medullary ACE activity. This change was due not to a modified affinity of ACE for the substrate but to an alteration in ACE maximal velocity or number of available molecules. UADX had no effect on adrenocortical ACE activity. When UADX was combined with right splanchnic denervation, the increase in adrenomedullary ACE activity was blocked. The results support the existence of a functional ACE in adrenal medulla that is under neuronal control.

Adrenal Medulla↗

A trans-acting mechanism represses the expression of the major transplantation antigens in mouse hybrid thymoma cell lines.

We have fused an H-2- thymoma (BM5R.9) with an H-2+ thymoma (BW5147) and have found that many of the resulting hybrids exhibit an H-2- phenotype. In several hybrids that were analyzed in detail, this phenotype is related to the absence of steady-state H-2 mRNA and shows some instability, possibly related to the loss of chromosomes in segregants. We conclude from our studies that BM5R.9 cells display a trans-acting mechanism that can repress the expression of H-2 antigens, and that the gene(s) causing the repression are not located on chromosome 17. This mechanism is not sufficient to explain the H-2- phenotype of BM5R.9, for which an additional, cis-acting process, must be postulated. We discuss these results in the context of the regulation of expression of the major class I transplantation antigens.

Animals↗

Adenovirus E1A-mediated regulation of class I MHC expression.

Expression of class I MHC transplantation antigens has been shown to be reduced in baby rat kidney (BRK) cells transformed by highly oncogenic adenovirus type 12 (Ad12), as compared with untransformed cells and cells transformed by non-oncogenic Ad5. Here we show that this reduction of class I expression also occurs in a variety of other primary cell cultures transformed by Ad12, and that reduction of class I gene expression occurs for all class I loci. Transfection of Ad5E1 into class I-negative Ad12-transformed BRK cells leads to complete restoration of class I expression. Introduction of Ad12E1 into most class I-positive established cell lines does not result in suppression of class I expression. However, transfection of the Ad12E1A region into a class I-positive cell line which was immortalized by a mutant Ad12E1A region resulted in suppression of class I gene expression, implying that the suppression of class I activity in Ad12-transformed cells is due to an active switching-off process.

Adenovirus Early Proteins↗

Diuretic and natriuretic action of rat atrial natriuretic peptide (6-33) administered intracerebroventricularly in rats.

Intracerebroventricular (IVT) administration of rat atrial peptide (6-33) (rANP) to conscious male hydrated or salt-loaded rats, resulted in significant increase in urinary volume. The diuretic effects of rANP occurred during the 3 hr period of urine collection and were most effective during the first hour. Most remarkably, rANP given IVT produced a dose-related increase in urinary sodium excretion at 3 hr. With a high dose of rANP, kaliuresis was significant only at 3 hr. Our results strongly suggest that ANF may play a significant role in central regulation of fluid homeostasis, and that its natriuretic and diuretic effects may be, at least in part, centrally mediated.

Animals↗

Photosynthesis of Ulva sp: III. O(2) Effects, Carboxylase Activities, and the CO(2) Incorporation Pattern.

Ulva, a common green seaweed, performs at the biochemical level as a typical C(3) plant. Over 90% of label was found in glycerate 3-phosphate following a 3 second (14)C pulse in the light, and the label was subsequently transferred to sugars. Also, the level of ribulose-1,5-bisphosphate carboxylase activity in crude extracts was about 10 times higher than that of phosphoenolpyruvate carboxylase. Concerning gas exchange, photosynthetic rates of Ulva showed no O(2) sensitivity, indicating that photorespiratory CO(2) losses are repressed as in C(4) plants. This apparent anomaly could be explained by the efficient HCO(3) (-) uptake system of Ulva which might concentrate CO(2) to the chloroplasts, thus suppressing the oxygenase activity of ribulose-1,5-bisphosphate carboxylase.

Journal Article↗

Increased number of angiotensin II binding sites determined by autoradiography in anterior pituitary of water-deprived and Brattleboro rats.

Pituitary angiotensin II (ANG) binding sites were characterized by autoradiography in individual male Long Evans (LE) rats, heterozygous Brattleboro (HZ) rats, and homozygous Brattleboro (DI) rats which were water satiated or water deprived. An additional group of DI rats was treated with arginine vasopressin for 1 week. The technique utilized 8-micron pituitary sections which were incubated with 125I-[Sar1]-ANG in concentrations ranging from 25 pM to 10 nM. Angiotensin binding in the anterior pituitary of water-satiated LE rats was characterized by a single class of high-affinity, saturable sites with a Bmax of 1.309 +/- 119 fmol/mg of protein and a Ka of 0.51 +/- 0.03 X 10(9) M-1. Compared to LE rats the density of ANG-binding sites in anterior pituitary was higher in DI rats (+84.2%), with HZ having an intermediate concentration (+41.6). Dehydration increased the ANG-binding site density in all groups. Five days of water deprivation increased the number of ANG-binding sites by 86.5% and by 36.9% in LE and HZ rats, respectively, when compared to their water-satiated controls. After 1 day of water deprivation, the ANG-binding site density increased by 19.4% in DI rats. No changes in ANG-binding sites occurred after hormonal replacement with arginine vasopressin in water-satiated DI rats. The binding affinity constant of the agonist for the ANG-binding sites compared with LE rats was decreased in both HZ and DI rats (-27.5 and -19.4%) and was also decreased after 5 days of dehydration in LE rats (-34%) when compared to the water-satiated state.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗

Decreased number and affinity of rat atrial natriuretic peptide (6-33) binding sites in the subfornical organ of spontaneously hypertensive rats.

Binding sites for rat atrial natriuretic peptide (6-33) were quantified by incubation of brain sections with (3-[125I]iodotyrosyl28) rat atrial natriuretic peptide (6-33), followed by autoradiography with computerized microdensitometry. Spontaneously hypertensive rats present lower numbers and lower affinity of binding sites than normotensive controls, Wistar-Kyoto rats, in the subfornical organ (binding capacity 61.7 +/- 8.9 and 124.3 +/- 10.7 fmol/mg protein; affinity constant 4.25 +/- 0.55 and 11.10 +/- 1.67 X 10(9) M-1, respectively). In the choroid plexus, hypertensive rats have lower numbers of sites than normotensive rats (binding capacity 72.7 +/- 10.5 and 173.6 +/- 22.8 fmol/mg protein, respectively), but there was no difference in the binding affinity (affinity constant 6.28 +/- 0.82 and 7.60 +/- 2.06 X 10(9) M-1, respectively). Our results suggest that discretely localized brain binding sites for rat atrial natriuretic peptide (6-33) may have a physiological function in genetically hypertensive rats.

Animals↗

Increased atrial natriuretic peptide (6-33) binding sites in the subfornical organ of water deprived and Brattleboro rats.

Binding sites for rat atrial natriuretic peptide (6-33) (ANP) were quantitated in the subfornical organ of chronically dehydrated homozygous Brattleboro rats unable to synthesize vasopressin; heterozygous Brattleboro rats, their controls, Long Evans rats and Long Evans rats after 4 days of water deprivation. Brain sections were incubated in the presence of 125I-ANP and the results analyzed by autoradiography coupled to computerized microdensitometry and comparison to 125I-standards. Brattleboro rats and water deprived Long Evans rats presented a higher number of ANP binding sites than their normally hydrated controls. Our results suggest a role of ANP binding sites in the subfornical organ in the central regulation of fluid balance and vasopressin secretion.

Animals↗

Pindolol decreases plasma angiotensin-converting enzyme activity in young spontaneously hypertensive rats.

The beta-adrenoceptor antagonist pindolol, given p.o. (10 mg/kg, once a day for 7 days) to 4 and 24 week old male spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY), decreased plasma angiotensin-converting enzyme (ACE) activity only in young SHR. Kinetic studies indicated that the treatment reduced the ACE maximal velocity or the number of available ACE molecules. In vitro studies showed that pindolol had no direct effects on the activity of plasma ACE. A decrease in soluble (loosely bound) ACE activity was observed in the lung of young SHR treated with pindolol. The results suggest that pindolol affects the site of origin of plasma ACE. The susceptibility of young SHR to pindolol may indicate a probable role of beta-adrenoceptors in the regulation of ACE activity in spontaneous (genetic) hypertension.

Animals↗

Functional analysis of the mouse H-2Kb gene promoter in embryonal carcinoma cells.

Mouse embryonal carcinoma (EC) cells do not express the major H-2 class I transplantation antigens. The latter, however, become detectable upon in vitro differentiation of EC cells. Neither class I H-2 genes nor the gene coding for beta-2 microglobulin (beta 2m) is transcribed in EC cells. We have constructed two hybrid plasmids containing the 5' flanking region of an H-2Kb gene followed by the coding regions of either the herpes simplex virus thymidine kinase (H-2 tk) or the chloramphenicol acetyl transferase (H-2 CAT) genes. Upon transfer into EC cells, the H-2 tk hybrid gene is expressed in F9 tk- cell lines which thus acquire a stable tk+ phenotype. When such transformed clones are induced to differentiate in vitro, tk activity shows a moderate increase, which reflects an increase in transcription of the hybrid gene. In transient transformation experiments, EC cells were found to express the H-2 CAT hybrid gene as well. We conclude that the 2 kilobase pair region of the H-2Kb gene which we used contains an active promoter region, but does not include all the elements required for the correct regulation of the H-2Kb gene.

Acetyltransferases↗

Asynchronous regulation of mouse H-2D and beta-2 microglobulin RNA transcripts.

The major transplantation (or H-2) antigens in the mouse are cell-surface glycoproteins composed of a heavy chain and a light chain, the beta-2 microglobulin (beta 2m). The expression of these proteins is regulated during development. Embryonic cells at early stages of development do not express these proteins. On the other hand, these molecules are present on the surface of all adult somatic cells. We investigated whether the expression of both chains was coordinately regulated. Using specific single-stranded DNA probes in an S1 nuclease analysis, we compared the relative amounts of H-2D and beta 2m transcripts in normal tissues, in transformed cells, and during embryonic development. Our results show that (1) the steady state level of beta 2m transcripts varies from one adult organ to another, while that of H-2D transcripts stays approximately the same; (2) upon transformation, the amount of H-2D-specific mRNA increases drastically, while the beta 2m mRNA level remains constant; (3) whereas the quantity of beta 2m mRNA increases during early development, the amount of H-2D mRNA remains at a very low level. These data suggest that the regulation of H-2D and beta 2m genes are not identical and that their activation during development is not synchronous.

Aging↗

Quantitative autoradiographic characterization of receptors for angiotensin II and other neuropeptides in individual brain nuclei and peripheral tissues from single rats.

Autoradiographic techniques coupled with computerized microdensitometry and comparison with 125I standards were used to characterize and quantitate receptors for neuropeptides in rat brain and adrenal and pituitary glands. These techniques are rapidly performed, anatomically precise, and more sensitive than membrane binding techniques. They permit the determination of complete saturation curves and Scatchard analysis in discrete nuclei of the rat brain and in single rat pituitary and adrenal glands. Angiotensin II (AII) receptors were quantitated after incubation of 16-micron tissue sections with the AII agonist 125I-[Sar1]-AII. High-affinity, high-density AII receptors were present in the organon subfornicalis, organon vasculosum laminae terminalis and nuclei triangularis septalis, suprachiasmatis, and paraventricularis of the rat and in rat adrenal capsule-zona glomerulosa area, adrenal medulla, and anterior pituitary. These techniques could be used for precise localization and quantitation of other neuropeptide receptors in single rat brain nuclei, after optimizing the assay conditions and provided that suitable 125I ligands are available.

Adrenal Cortex↗