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Biomedical subjects

W C Hubbard

Publications and source records attributed to W C Hubbard.

104 records · Page 6Linked to original sources
PubMed1980

The site of VX2 tumor transplantation affects the development of hypercalcemia in rabbits.

The relationship between plasma levels of 15-keto-13, 14-dihydroprostaglandin E2 (15K-H2-PGE2) and serum calcium levels was studied in nontumor-bearing rabbits and in rabbits bearing the VX2 carcinoma intramuscularly and intra-abdominally. The plasma levels of 15K-H2-PGE2 in the two groups of tumor-bearing animals did not vary significantly but was several fold greater than in nontumor-bearing rabbits. Rabbits bearing the VS2 carcinoma intramuscularly developed hypercalcemia between the second and third week after implantation of neoplastic tissue and remained hypercalcemic until they expired. The serum calcium levels in rabbits bearing the VX2 carcinoma intra-abdominally did not vary significantly from those of nontumor-bearing rabbits. The differences in the serum calcium levels in rabbits bearing the VX2 carcinoma at intramuscular and intra-abdominal implant sites may be related to different extents of metabolism by the lung and by the liver of prostaglandin E2 or other cyclooxygense of polyenoic fatty acids produced by the tumor.

Animals↗
PubMed1980

Preparation and characterization of hydroperoxy-eicosatetraenoic acids (HPETEs).

5-, 8-, 9-, 11-, 12- AND 15 hydroperoxy-eicosatetraenoic acids (HPETEs) were generated from arachidonic acid by a reaction with H2O2 and Cu++ ions. They were purified by high performance liquid chromatography, either on a silica gel (mu Porasil) column or on a reversed phase (mu Bondapak C18) column. The yield of 5-HPETE was considerably greater when the mu Bondapak C18 column was used. The HPETEs were characterized and assayed by their ability to oxidize triphenylphosphine: triphenylphosphine oxide formation was monitored by gas chromatography. When stored in methylene chloride at -20 degrees C, the HPETES were stable for several months.

Arachidonic Acids↗
PubMed1980

Modulation of human neutrophil function by monohydroxy-eicosatetraenoic acids.

The generation from arachidonic acid and purification of large quantities of a series of monohydroxy-eicosatetraenoic acids (HETEs) which differed only in the position of the hydroxyl group permitted an in vitro analysis of the relative effects of the HETEs on a variety of human neutrophil functions. All of the HETEs elicited maximal neutrophil chemotactic responses of comparable magnitude, but the chemotactic potencies exhibited a distinct rank order with 5-HETE greater than 8-HETE:9-HETE (85:15, w:w) greater than 11-HETE=12-L-HETE. Peak chemotactic responses were achieved at concentrations of 1 microgram/ml for 5-HETE, 5 microgram/ml for 8-HETE:9-HETE and 10 microgram/ml for 11-HETE and 12-L-HETE. In the absence of a concentration gradient, the HETEs were similar in potency with respect to the stimulation of neutrophil chemokinesis and the enhancement of the expression of neutrophil C3b receptors. At optimally chemotactic and chemokinetic concentrations, none of the HETEs stimulated the generation of superoxide by neutrophils, altered the expression of neutrophil IgG-Fc receptors, or evoked the release of lysosomal enzymes. Methyl esterification of 5-HETE and 12-L-HETE reduced the chemotactic activity to less than 12% of that of the parent compound. The HETE methyl esters competitively inhibited the chemotactic activity of the homologous free acids by approximately 50% at equimolar concentrations, without inhibiting the chemotactic activity of formyl-methionyl peptides or of chemotactic fragments of the fifth component of complement (C5fr). The stimulus specificity of the competitive inhibition of chemotaxis by HETE methyl esters and the functional selectivity of the HETEs as compared to the formyl-methionyl peptides and C5fr, which stimulate neutrophil oxidative metabolism and lysosomal enzyme release, suggest that HETEs activate human neutrophils by a unique mechanism.

Arachidonic Acids↗
PubMed1979

Uterine vein prostaglandin levels in late pregnant dogs.

We studied the uterine venous plasma concentrations of prostaglandins E2, F2 alpha, 15 keto 13,14 dihydro E2 and 15 keto 13,14 dihydro F2 alpha in late pregnant dogs in order to evaluate the rates of production and metabolism of prostaglandin E2 and F2 alpha in pregnancy in vivo. We used a very specific and sensitive gas chromatography-mass spectrometry assay to measure these prostaglandins. The uterine venous concentrations of prostaglandin E2 and 15 keto 13,14 dihydro E2 were 1.35 +/- .27 ng/ml and 1.89 +/- .37 ng/ml, respectively; however, we could not find any prostaglandin F2 alpha and very little of its plasma metabolite in uterine venous plasma. Since uterine microsomes can generate prostaglandin F2 alpha and E2 from endoperoxides, prostaglandin F2 alpha production in vivo must be regulated through an enzymatic step after endoperoxide formation. Prostaglandin E2 is produced by pregnant canine uterus in quantities high enough to have a biological effect in late pregnancy; however, prostaglandin F2 alpha does not appear to play a role at this stage of pregnancy.

Animals↗
PubMed1979

The role of renal metabolism of PGE2 in determining its activity as a renal vasodilator in the dog.

Since the mammalian renal cortex avidly metabolizes prostaglandin E2 (PGE2), we examined the importance of renal metabolism of PGE2 in determining its renal vascular activity in the dog. We used 13, 14 dihydro PGE2 (DHPGE2) as a model compound to study this because DHPGE2 retains similar activity to the parent prostaglandin, PGE2, but is a poorer substrate than PGE2 for both the metabolism and the cellular uptake of the prostaglandins. Using dog renal cortical slices, we found that under similar experimental conditions, PGE2 was metabolized several-fold faster than DHPGE2. Both prostaglandins were metabolized to the 15 keto 13, 14 dihydro PGE2, which was positively identified using GC-MS. In vivo, we infused increasing concentrations of DHPGE2 into the renal artery of dogs and measured renal hemodynamic changes using radioactive microspheres. DHPGE2 was a potent renal vasodilator beginning at an infusion rate of 10(-9)g/kg/min. When compared to PGE2, DHPGE2 was about 10 times more potent in affecting renal vasodilation. The intrarenal redistribution of blood flow towards the inner cortex seen with DHPGE2 was identical to that seen with PGE2. We conclude that renal catabolism of PGE2 is very important in limiting the in vivo biological activity of PGE2, but regional differences in metabolism of PGE2 within the cortex are an unlikely determinant of the pattern of redistribution of renal blood flow.

Animals↗
PubMed1979

Relative vasodilator potency of ATP, ADP, AMP and adenosine (Ado) in the cat hind limb.

The relative vasodilator potencies of ATP, ADP, AMP and Ado were determined by bioassay using the skinned and unskinned cat hind limb and comparing the increase in blood flow induced by intra-arterial injection of the adenyl compounds. The relative vasodilator potencies of these agents were similar in either hind limb preparation and were as follows: ADP greater than ATP greater than Ado greater than or equal to AMP. ADP was 2-4 times more potent than ATP and about 200 times more potent than Ado or AMP. When the adenyl compounds were incubated in blood for 2 min before injection, their potencies were not changed in the bioassay. Furthermore, in a chemical assay, after Ado and ADP were incubated for 2 min in cat blood (at the concentrations used in the bioassay) only 3% of Ado and 29% of ADP were metabolized. The results indicate that ATP and ADP exert a vasodilator effect that is not dependent upon their conversion to Ado. This deduction cannot be made for AMP because its relative potency was quite similar to Ado. Since a similar relative potency was found in the skinned and the unskinned hind limb, it is concluded that the vasodilator effect due to ATP and ADP is occurring in skeletal muscle but this does not exclude simultaneous cutaneous vasodilation. The vasodilator effect of adenyl compounds may be related to purinergic receptor activation.

Adenine Nucleotides↗
PubMed1978

The lack of an effect of furosemide on uterine prostaglandin metabolism in vivo.

We determined the effect of 2 mg/kg intravenous furosemide on the production and metabolism of prostaglandin E2 in the utero-placental unit of pregnant dogs. Uterine venous prostaglandins E2 and 15-keto-13,14-dihydro E2 were measured by gas chromatography-mass spectrometry. Even though the dose of furosemide was adequate to effect a good diuresis, neither the production nor the metabolism of prostaglandin E2 by the uterus was altered by that dose of the drug. Using radioactive microspheres to measure hemodynamic parameters, we observed no change in uterine vascular resistance while renal vascular resistance decreased. Although the renal concentration of furosemide may be higher than the uteroplacental concentration, there is so far no evidence in vivo that usual doses of furosemide enhance the production or inhibit the metabolism of prostaglandin E2.

Animals↗
PubMed1978

Indomethacin is a placental vasodilator in the dog. The effect of prostaglandin inhibition.

The effect of 8 mg/kg of indomethacin on uterine blood flow, prostaglandin production, and intraamniotic fluid pressure was examined in late pregnant dogs. Uterine blood flow was measured with 15 mum radiolabeled microspheres. Because we found that a significant percentage of the microspheres shunted through the placental circulation into the lungs, we calculated placental blood flow by adding the shunted microspheres through the placenta to the nonshunted microspheres in the placenta. Total uterine blood flow significantly increased from 271+/-69 ml/min during control period to 371+/-72 ml/min (P < 0.01) 30 min after indomethacin. This increase was attributable to the change in blood flow to the placental circulation (222+/-58 to 325+/-63 ml/min; P < 0.01). Associated with these hemodynamic changes we found an almost complete suppression of uterine prostaglandin E(2) production (1,654+/-305 to 51+/-25 pg/ml; P < 0.01) as measured by gas chromatography-mass spectrometry. In addition, we found that indomethacin treatment resulted in uterine relaxation as measured by intraamniotic fluid pressure changes (11.2+/-1.3 mm Hg to 8.5+/-1.2 mm Hg; P < 0.001). We conclude that indomethacin causes an increase in placental blood flow without any change in flow to the rest of the uterus, and that this dose of the drug inhibits greater than 95% of uterine prostaglandin production. In addition, indomethacin is responsible for uterine relaxation. The increase in placental blood flow after indomethacin is probably a result of uterine relaxation, which is secondary to prostaglandin synthesis inhibition.

Amniotic Fluid↗
PubMed1977

Prostaglandin-mediated hypercalcemia in the VX2 carcinoma-bearing rabbit.

Prostaglandin biosynthesis and metabolism were studied in the VX2 carcinoma-bearing rabbit, an animal model of prostaglandin-mediated hypercalcemia. All the identification and quantification of the prostaglandins were done by gas chromatography-mass spectrometry. The tumor incubated in vitro converted exogeneous arachidonic acid principally to PGE2. Biosynthesis from endogenous precursor lipids yields mainly PGE2 and PGF2alpha. The 100,000 x g supernatant fluid of the tumor did not contain any metabolizing enzymes. Significant hypercalcemia developed between the first and second week after tumor implantation. The levels of the major plasma metabolite of PGE2, 15-keto-13,14-dihydro-PGE2, became elevated at one week, had risen 25-fold by the end of the second week, and at the fourth week were elevated to 256 times the pre-incubation levels. The concentration of 15-keto-13,14-dihydro-PGF2alpha in plasma rose in parallell but to a lesser degree. 7alpha-hydroxy-5,11-diketotetranor-prostane-1,16-dioic acid, the major urinary metabolite of the E prostaglandins, was elevated two weeks after tumor implantation and rose until the fifth week. Indomethacin treatment lowered both serum calcium and the plasma level of 15-keto-13,14-dihydro-PGE2.

Animals↗
PubMed1976

Determination of 15-keto-13,14-dihydro-metabolites of PGE2 and PGF2alpha in plasma using high performance liquid chromatography and gas chromatography-mass spectrometry.

A method is described for the measurement of 15-keto-13,14-dihydrometabolites of PGE2 and PGF2alpha in peripheral human plasma. This involves purification by high performance liquid chromatography followed by determination of levels by combined gas chromatography-mass spectrometry using tetradeuterated analogs of the metabolites as internal standards. The levels of these metabolites in plasma are considered to be a more reasonable index of the entry of PGE2 and PGF2alpha into peripheral blood than are the levels of the corresponding primary prostaglandins. The endogenous levels of 15-keto-13, 14-dihydro-PGE2 and 15-keto-13,14-dihydro-PGF2alpha found in peripheral plasma are 33 +/- 10 pg/ml (SD; n=6) and 40 +/- 16 pg/ml (SD; n=6), respectively.

Chromatography, Gas↗
PubMed1975

Dissociation constant of the norepinephrine-receptor complex in normotensive and hypertensive rats.

Previous reports have suggested that smooth muscle obtained from the thoracic aorta of spontaneously hypertensive rats is less responsive to vasoconstrictive agents than that obtained from normotensive rats. The present study was undertaken to determine whether the responsiveness of aortic muscles from normotensive and spontaneously hypertensive rats correlates with a difference in the affinity of the adrenergic receptors for norepinephrine and whether antihypertensive therapy alters the affinity of the adrenergic receptors for norepinephrine. The affinity of the adrenergic receptors for norepinephrine was determined by computing the dissociation constant of the norepinephrine-receptor complex (KDR). The values computed for KDR in aortic muscles from normotensive and spontaneously hypertensive rats that had received no antihypertensive therapy were 1.07 X 10(-7)M and 1.17 X 10(-7)M, respectively. The values computed for KDR in aortic muscles from normotensive and spontaneously hypertensive rats that had received antihypertensive therapy were 1.38 X 10(-7)M and 1.29 X 10(-7)M, respectively. The differences in these values for KDR are not significant. These results indicate that the difference in the contractility of aortic muscles from normotensive and spontaneously hypertensive rats is not related to an alteration in the affinity of the adrenergic receptors for norepinephrine and that the affinity of the adrenergic receptors for norepinephrine is not altered by antihypertensive therapy. Thus, it appears that the etiology of hypertension cannot be directly correlated with a difference in the affinity of the adrenergic receptors for norepinephrine.

Animals↗