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P Needleman

Publications and source records attributed to P Needleman.

At least 55 records · Page 3Linked to original sources
PubMed1990

Augmentation of the natriuretic activity of exogenous and endogenous atriopeptin in rats by inhibition of guanosine 3',5'-cyclic monophosphate degradation.

To investigate the relationship between AP, cyclic GMP, and sodium excretion, we studied the effect of a cyclic GMP phosphodiesterase inhibitor (M + B22948) on the natriuretic response to (a) an infusion of AP (103-126) and (b) acute volume expansion in rats. The phosphodiesterase inhibitor markedly potentiated the effect of low-dose AP infusions on urinary sodium and cyclic GMP excretion without potentiating the fall in blood pressure. Acute volume expansion (1% body wt) led to small but significant (P less than 0.01) rises in plasma AP and urinary cyclic GMP levels. Pretreatment with the phosphodiesterase inhibitor enhanced the natriuretic and cyclic GMP response to volume loading, an effect that was attenuated by administration of a monoclonal antibody directed against AP. These data indicate that cyclic GMP mediates the natriuretic activity of AP and AP and cyclic GMP play active roles in the natriuresis of acute volume expansion. Moreover, pharmacological manipulation of cyclic GMP levels may prove a useful therapeutic strategy for facilitating the natriuretic but not the hypotensive effects of AP.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1990

Selective regulation of cellular cyclooxygenase by dexamethasone and endotoxin in mice.

We have studied the effect of glucocorticoids administered in vivo on the activity and synthesis of the cyclooxygenase enzyme (COX) in mice treated with or without concurrent intravenous administration of LPS. Mouse peritoneal macrophages from LPS-treated animals showed a two to three fold increase in COX activity determined by the production of PGE2 and PGI2 after stimulation of the cells with exogenous arachidonate. Dexamethasone injected simultaneously with LPS, 12 h before killing of the animal and removal of the macrophages, completely blocked the LPS-induced increase COX activity in peritoneal macrophages. The regulation observed in COX activity by LPS and dexamethasone are due primarily to changes in COX mass as determined by immunoprecipitation of [35S]methionine endogenously labeled enzyme. In contrast, the COX present in the nonadherent cells and in renal medullary microsomes obtained from the same animals, showed no significant changes between treatments. These results indicate that LPS in vivo stimulates COX synthesis in the peritoneal macrophages but not in the kidney. The effect of dexamethasone to inhibit COX synthesis is selective to the LPS-induced enzyme.

Animals↗
PubMed1990

Alternative mechanisms for atriopeptin prohormone processing by isolated perfused rat hearts.

The isolated perfused rat heart releases atriopeptin-28 [AP28 (ANF99-126)], whereas the storage form of AP in the heart is the intact prohormone AP126 (ANF1-126). Right atrial stretch or phenylephrine (5 x 10(-5) M) stimulated the release of AP28. The processing of the prohormone during stretch was inhibited by infusion of the protease inhibitor aprotinin, resulting in the appearance of intact AP126 in the cardiac effluent. Other protease inhibitors including p-aminobenzamidine and soybean trypsin inhibitor did not alter prohormone processing by the isolated heart subjected to stretch. In contrast, aprotinin did not block the prohormone processing induced by phenylephrine. Ca+(+)-free medium markedly inhibited prohormone processing during stretch without a significant effect on AP release, whereas phenylephrine-stimulated AP release was completely suppressed by Ca+(+)-free medium. Exogenous AP126 could be cleaved by isolated rat hearts perfused either with Krebs-Henseleit solution or with Ca+(+)-free medium. However, amino acid sequence analysis revealed that the prohormone cleavage in Ca+(+)-free medium occurred at sites other than between Arg98 and Ser99 and that the resultant low molecular weight APs were not AP28. These findings suggest: 1) the characteristics of the enzyme(s) involved in the processing of AP prohormone in isolated perfused rat hearts are different from the described properties of purified enzymes; 2) in isolated perfused rat hearts the specific AP processing enzyme is Ca++ dependent, whereas nonspecific cleavage does not necessarily require Ca++ and 3) two independent AP processing pathways differentially activated by mechanical (stretch) and pharmacologic (alpha 1-adrenergic agonist) stimuli exist.

Animals↗
PubMed1989

Brain natriuretic peptides: differential localization of a new family of neuropeptides.

Brain natriuretic peptide (BNP) is a recently discovered neuropeptide, isolated from the porcine brain, that is highly homologous to atriopeptin (AP), the atrial natriuretic peptide. We used a set of highly selective antisera against the two peptides to map their differential distribution immunohistochemically in the rat central nervous system. BNP immunoreactivity has a distinct distribution, involving many central autonomic and endocrine control structures that contain little if any AP immunoreactivity. AP and BNP belong to a family of neuropeptides that may be important in central cardiovascular control.

Animals↗
PubMed1989

The cell biology of fibroblast cyclooxygenase.

We have prepared polyclonal antisera against sheep seminal vesicles cyclooxygenase (COX) which cross-reacted with human COX. We employed this antisera in studies with human dermal fibroblast cultures to immunoprecipitate selectively the COX enzyme. Labeling of the cells with [35S]-methionine, solubilization of cellular COX followed by its immunoprecipitation, SDS-PAGE electrophoresis and fluorography enabled us to determine directly the synthetic rate of COX protein and its modulation by the monokine interleukin-1 (IL-1). The immunoprecipitated [35S]-labeled COX, as judged from SDS-PAGE electrophoresis, has a molecular size of approximately 73,000 daltons, similar to that of native sheep COX and [3H]-acetyl COX. IL-1 stimulation of enhanced COX synthesis was time and dose dependent; as little as 0.03 units/ml of IL-1 produced significant stimulation of [35S]-labeled COX synthesis. Maximum stimulation was 3-10-fold after preincubation of the cells with IL-1 for 12-16 hours. IL-1 treatment of cells in serum-free media yielded parallel dose response curves for stimulation of PGE2 formation, cellular solubilized COX activity and synthesis of newly formed COX, suggesting that this IL-1 effect is mediated solely via induction of new COX protein synthesis. In contrast, IL-1 effect on cells incubated in the presence of fetal calf serum is more complex. Serum synergistically augments the IL-1 effect on PGE2 synthesis in intact cells but concurrently blunts IL-1 induction of COX synthesis, thus suggesting that a factor (or factors) in serum may stimulate PGE2 production by activating cellular phospholipase(s).

Animals↗
PubMed1989

Essential fatty acid deficiency inhibits early but not late leukocyte infiltration in rabbit myocardial infarcts.

Essential fatty acid (EFA) deficiency, induced by elimination of the dietary (n-6) fatty acids, has been shown to limit inflammatory cell influx and consequent enhanced eicosanoid production in experimental glomerulonephritis and hydronephrosis. To determine whether EFA-deficiency exerts anti-inflammatory effects following left ventricular myocardial infarction (LVMI), male weanling rabbits were fed EFA-deficient diet for 3 months prior to 60 minutes of distal left circumflex coronary artery occlusion followed by reperfusion. One and 4 days later, corresponding to infiltration of cardiac tissue with polymorphonuclear (PMN) and mononuclear leukocytes respectively, infarcted hearts were buffer perfused and stimulated to produce eicosanoids with f-met-leu-phe or bradykinin. One day following LVMI, the hearts of EFA-deficient rabbits demonstrated a marked suppression of PMN infiltration and eicosanoid production relative to controls. Four days following myocardial infarction, no differences were observed in mononuclear cell invasion, collagen deposition, or eicosanoid production between EFA-deficient and normal hearts. Our data show that EFA-deficiency inhibits PMN influx and consequent enhanced eicosanoid production without affecting the later appearance of mononuclear cells, collagen deposition, or eicosanoid production. Recent studies have shown that suppression of PMN invasion limits the extent of tissue damage following LVMI. Selective inhibition of PMN infiltration is possible and may be useful in the management of acute myocardial infarction.

6-Ketoprostaglandin F1 alpha↗
PubMed1989

Differentiation in vivo of the sequential blood cell invasion following ureter obstruction of the rabbit kidney.

Inflammatory cellular changes occurring during renal injury as a consequence of unilateral ureter obstruction were studied in vivo using 111In-labeled leukocytes and 111In-labeled platelets. The radiolabeled cells were administered at various times after induction of hydronephrosis by unilateral ureter ligation in New Zealand rabbits. The contralateral kidney (CLK) served as the control. The migration and accumulation of the radiolabeled cells was monitored by dual isotope scintigraphic imaging (i.e. 99mTc-labeled red blood cell subtraction) and confirmed by direct analysis of removed epsilateral hydronephrotic kidney (HNK) and CLK at the time of sacrifice. When 111In platelets were administered, significantly greater uptake in the hydronephrotic kidney was observed within 15 min of ureter ligation. Furthermore, the HNK accumulated twice the amount of 111In radioactivity than the CLK, when either the polymorphonuclear neutrophils (PMN) or monocytes were injected at the time of ureter obstruction. The differences between HNK and CLK were scintigraphically detectable between 6 and 12 h after hydronephrosis respectively; and 111In-labeled monocytes selectively accumulated in HNK if administered during the first 48 h after ureter ligation. These data suggest that inflammatory cells sequentially accumulate following ureter obstruction and are derived from circulating white blood cells.

Animals↗
PubMed1989

Temporal and pharmacological division of fibroblast cyclooxygenase expression into transcriptional and translational phases.

We have recently shown that the synthesis of cyclooxygenase [also called prostaglandin (PG) synthase or PG endoperoxide synthase; 8,11,14-icosatrienoate, hydrogen-donor:oxygen oxidoreductase, EC 1.14.99.1] in human dermal fibroblasts is markedly stimulated by the cytokine interleukin 1 (IL-1). We now show that the temporal sequence of the induced synthesis of PG synthase can be separated into an early transcriptional (i.e., actinomycin D inhibitable) phase and a subsequent translational (cycloheximide but not actinomycin D inhibitable) phase and that IL-1 exerts its effect during the transcriptional phase. Phorbol 12-myristate 13-acetate also stimulates synthesis of PG synthase and, together with IL-1, produces a synergistic stimulatory effect. Inhibitors of protein kinase C activation abolished the stimulatory effect of IL-1, suggesting that protein kinase C activation is a critical event in the signal-transduction sequence of the IL-1-induced increase of PG synthase synthesis. The antiinflammatory glucocorticosteroids dexamethasone and triamcinolone, but not progesterone or testosterone, were potent inhibitors of PG synthase synthesis (complete inhibition at 20 nM; IC50, 1 nM) when added during the translational phase of the synthesis sequence. The glucocorticosteroid effect was blocked by RNA and protein synthesis inhibitors. This report suggests that glucocorticosteroids exert their effect via a newly synthesized protein, causing a profound translational control of PG synthase synthesis. This novel mechanism of suppression of arachidonate metabolism is distinct from any influence of steroids on phospholipase A2 activity.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1989

Mechanisms underlying atriopeptin-induced increases in hematocrit and vascular permeation in rats.

Infusion of atriopeptin into humans and animals induces diuresis, natriuresis, hemodynamic changes, and an increase in arterial hematocrit. The objective of the present study was to elucidate the mechanism(s) responsible for the increase in hematocrit in rats given atriopeptin-24 (AP-24). Infusion of AP-24 for 30 minutes increased large vessel and total vascular hematocrits by 10-15% while decreasing microvascular hematocrits by 9-26% in numerous tissues. Regional vascular permeation by [131I] bovine serum albumin was markedly increased (2-5.6-fold) in many tissues, consistent with a 16% decrease in plasma volume. AP-24 infusion had no effect on extracellular fluid volume or the volume of circulating red cells. Vascular resistance was decreased and was associated with a significant increase in blood flow in many, but not all, tissues. In the atrium and in the small and large intestine the percentage decrease in microvascular hematocrit exceeded the increase in blood flow. These observations indicate that the increase in large vessel hematocrit induced by AP-24 infusion 1) is accompanied by a decrease in (microvascular) hematocrit in many tissues, 2) reflects an increase in overall (i.e., total vascular hematocrit), and 3) is the consequence of a decrease in plasma volume resulting from a marked increase in the rate of vascular permeation by plasma constituents in multiple tissues.

Animals↗
PubMed1989

Role of invading leukocytes in enhanced atrial eicosanoid production following rabbit left ventricular myocardial infarction.

The isolated perfused hearts of rabbits previously subjected to in vivo left ventricular myocardial infarction (LVMI) show a 5-10-fold increase in f-Met-Leu-Phe (FMLP) and bradykinin (BK)-stimulated eicosanoid metabolite production relative to noninfarcted hearts. This exaggerated arachidonate metabolism has been shown to occur primarily in the cardiac atria, a site remote from the zone of injury and to be associated with a 10-15-fold increase in atrial FMLP receptor number in the absence of atrial inflammation. All of these changes were temporally related to leukocyte infiltration into the infarct zone. To determine whether invading leukocytes mediate these responses, acute inflammatory cell influx was suppressed either by inducing leukopenia with nitrogen mustard or by administration of BW-755C, a mixed cyclooxygenase-lipoxygenase inhibitor. Both pharmacological manipulations resulted in a decrease in inflammatory cells in the infarct zone and a marked suppression (50-70%) of ex vivo agonist-stimulated eicosanoid metabolite production from perfused hearts and isolated atria. These manipulations also resulted in reversal of ex vivo FMLP-induced coronary vasoconstriction as well as augmentation of BK-induced coronary vasodilation. Further studies in nitrogen mustard-treated animals revealed a suppression of the LVMI-stimulated increase in atrial FMLP receptor number. These data show that suppression of leukocyte invasion after LVMI attenuates enhanced cardiac and atrial eicosanoid metabolite production, and results in marked changes in coronary vascular reactivity. An additional finding was that basal and stimulated LTB4 production was markedly increased in infarcted hearts. In vivo suppression of the increase in LTB4 production by BW-755C was associated with inhibition of inflammatory cell influx into the infarct zone. It therefore appears that LTB4 may be an important proinflammatory mediator of leukocyte invasion after LVMI.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗
PubMed1989

Stretch-induced atriopeptin secretion in the isolated rat myocyte and its negative modulation by calcium.

Cellular mechanism(s) regulating atriopeptin secretion and processing by the atrial myocyte are currently unknown. Osmotic stretch of isolated atrial myocytes as well as potassium chloride depolarization were potent stimuli of atriopeptin secretion. Release was potentiated by buffering either extracellular calcium with EGTA or intracellular calcium with the intracellular chelator, BAPTA AM. Atrial release of atriopeptin was inhibited after administration of ionomycin which elevates intracellular calcium. Fetal or early neonatal ventricular myocytes actively synthesize atriopeptin. Atriopeptin secretion by ventricular myocytes was also markedly potentiated by osmotic stretch as well as KCl depolarization. Only the 126 amino acid prohormone was secreted by the stretch-stimulated atrial and ventricular myocyte. These data suggest that stretch of the myocyte plasma membrane is a major stimulus for atriopeptin secretion and that atriopeptin secretion is not stimulated by raising intracellular calcium and appears to be negatively modulated by this cation. Like the atrial myocyte, the ventricular myocyte possesses the cellular mechanism(s) necessary to secrete atriopeptin by a regulated mechanism.

Animals↗
PubMed1989

Relationship between plasma atriopeptin concentration and function in the conscious primate.

The renal actions of atriopeptins (APs) 24, 21 and 28 were examined in the conscious primate, macaca fascicularis. AP-24 increased urine flow rate and sodium excretion 20- and 100-fold, respectively. The circulating form of the atriopeptins, AP-28, had similar, even slightly greater (25%) effects when compared to AP-24. AP-21 on the other hand had dramatically reduced effects, less than 20%, when compared to either AP-24 or AP-28. Infusion of AP-24 resulted in marked increases in plasma immunoreactive AP and in renal function. There were direct, significant linear relations between plasma levels and arterial pressure, heart rate, glomerular flow rate, urine flow rate, sodium and potassium excretion. However, the threshold for these effects was generally higher than expected, i.e., greater than 100 pg/ml. Interestingly, there was a 4-fold greater slope for sodium excretion when compared to other renal functions implying a distinctly different mechanism of action. Whereas, the plasma half-life of the peptide was 2 to 3 min, the biological half-life varied from 6 min for sodium excretion to 10 min for urine flow and potassium excretion. The increased slope for the relationship between sodium excretion and plasma AP concentration and the short half-life for sodium excretion indicate that the change in renal sodium handling is independent of urine flow rate and glomerular filtration rate. There is a direct and linear relationship between plasma peptides and renal function which may imply a cause and effect relationship. This extrapolation may, however, be valid only when plasma peptide levels are elevated markedly.

Animals↗
PubMed1989

Pharmacological manipulation of inflammation in rabbit hydronephrosis: effects of a combined cyclooxygenase/lipoxygenase inhibitor ethoxyquin, a thromboxane synthase inhibitor RS-5186 and a PAF antagonist L-659,989.

The rabbit hydronephrotic kidney (HNK) is a model of renal inflammation characterized by a marked increase in arachidonic acid metabolism that is temporally associated with an inflammatory cell influx into the injured tissue. The HNK exhibits an exaggerated elaboration of eicosanoids ex vivo in response to either bradykinin or the inflammatory cell agonist n-formyl-methionyl-leucyl-phenylalanine (fMLP) compared with the unobstructed contralateral kidney. To pharmacologically manipulate inflammatory cell influx into the HNK we administered ethoxyquin (200 mg/kg p.o.), a combined cyclooxygenase/lipoxygenase inhibitor, RS-5186 (10 mg/kg p.o.), a thromboxane synthase inhibitor or L-659,989 (5 mg/kg p.o.), a platelet activating factor antagonist, before and at various times during the development of hydronephrosis. Only ethoxyquin reduced inflammatory cell influx into the HNK and thereby prevented the enhancement of microsomal cyclooxygenase activity and attenuated the elaboration of eicosanoids ex vivo. Collectively, these results suggest a primary role of an eicosanoid, possibly leukotriene B4, but not thromboxane A2 or the chemotactic phospholipid, platelet activating factor, as a mediator of inflammatory cell influx resulting from ureter obstruction.

Animals↗
PubMed1988

Strategies for the in vivo inhibition of endogenous atriopeptin.

Pharmacological strategies have been employed to manipulate the response to atriopeptin (AP-28; ANF 99-126) in rats. These approaches include: heparin infusion which blocks the natriuresis-diuresis produced by exogenous AP infusion or by acute volume expansion; and autoimmune rats with endogenous antibody that blocks the natriuresis-diuresis produced by administration of exogenous AP or by acute volume expansion in anesthetized rats. Administration of the pressor analog 1-deamino-arg8-vasopressin (dAVP) to rats produces an elevation in systemic and right atrial blood pressure and markedly increases atriopeptin blood levels. The dAVP also produces a delayed natriuresis-diuresis which is effectively blunted in the autoimmune or heparin treated rats. The above approaches provide tools, in the absence of a specific receptor antagonist, with which to validate the intrinsic involvement of AP in physiological, pharmacological, or pathophysiological events.

Animals↗
PubMed1988

Macrophages isolated from liver granulomas of murine Schistosoma mansoni synthesize predominantly TxA2 during the acute and chronic phases of infection.

Macrophages isolated from liver granulomas of mice infected with Schistosoma mansoni for 8 or 20 wk synthesize predominantly thromboxane A2 with smaller amounts of the PGE2 and PGI2. There is no physiologic production of leukotrienes, as determined by RIA and HPLC. Thromboxane A2 is the predominant arachidonic acid metabolite whether the cells are stimulated by a phagocytic stimuli such as zymosan or the exogenous substrates arachidonic acid and PGH2. These data indicate that the predominant arachidonate enzymatic activity in these cells is thromboxane synthase.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗