Search PubMed⌕ Search

Biomedical subjects

J E Clark

Publications and source records attributed to J E Clark.

At least 55 records · Page 3Linked to original sources
PubMed1993

Mechanism-based inhibitors of prostaglandin omega-hydroxylase: (R)- and (S)-12-hydroxy-16-heptadecynoic acid and 2,2-dimethyl-12-hydroxy-16-heptadecynoic acid.

12-Hydroxy-16-heptadecynoic acid has been shown to selectively inactivate cytochrome P450 4A4, a pulmonary cytochrome P450 enzyme that catalyzes the omega-hydroxylation of prostaglandins [Muerhoff, A. S.; Williams, D. E.; Reich, N. O.; CaJacob, C. A.; Ortiz de Montellano, P. R.; Masters, B. S. S. J. Biol. Chem. 1989, 264, 749-756]. Potent, specific inhibitors of this enzyme are required to explore its physiological role. In a continuing effort to develop such agents, the two enantiomers of 12-hydroxy-16-heptadecynoic acid have been stereospecifically synthesized, their absolute stereochemistry confirmed, and the dependence of enzyme inactivation on absolute stereochemistry determined using cytochrome P450 4A4 purified from the lungs of pregnant rabbits. The 12S enantiomer is roughly twice as active (KI = 1.8 microM, t1/2 = 0.7 min) as the 12R enantiomer (KI = 3.6 microM, t1/2 = 0.8 min), but the chirality of the hydroxyl group is not a major determinant of the specificity for the prostaglandin omega-hydroxylase. The flexibility of the acyclic skeleton of the inhibitor may account for the relatively low enantiomeric discrimination. 2,2-Dimethyl-12-hydroxy-16-heptadecynoic acid, an analogue that cannot undergo beta-oxidation, has also been synthesized as a potential in vivo inhibitor of the enzyme and has been shown to inactivate the purified enzyme with KI = 4.9 microM and t1/2 = 1.0 min. These acetylenic agents, particularly the dimethyl analog, are promising in vivo inhibitors of cytochrome P450 4A4.

Animals↗
PubMed1993

Characterization of a cDNA encoding a human kidney, cytochrome P-450 4A fatty acid omega-hydroxylase and the cognate enzyme expressed in Escherichia coli.

A cDNA encoding a cytochrome P-450 4A (CYP4AII) was cloned from a human kidney cDNA library. Northern blot analysis and RNase protection assays indicate that related mRNAs occur in kidney and liver with the highest abundance found in kidney. The enzyme was expressed from its cDNA in Escherichia coli. A solubilized preparation of the enzyme reconstituted with cytochrome P-450 reductase catalyzed the omega-hydroxylation of lauric acid, palmitic acid, and arachidonic acid with turnover numbers of 9.8, 2.2 and 0.55 min-1, respectively. Little or no activity was detected toward prostaglandins A1 and E1.

Amino Acid Sequence↗
PubMed1993

Induction of flavin-containing monooxygenase (FMO B) in rabbit lung and kidney by sex steroids and glucocorticoids.

Modulation of lung and kidney levels of flavin-containing monooxygenase (FMO) isoform B following sc administration of dexamethasone, progesterone, beta-estradiol, aldosterone, or beta-estradiol plus progesterone was examined in male rabbits. Both dexamethasone and progesterone increased FMO B protein levels in lung. N-Oxygenation of N,N'-dimethylaniline (DMA) was significantly induced in lung microsomes from rabbits treated with progesterone. In kidney, only dexamethasone was effective in inducing FMO B protein levels and DMA N-oxygenation. Surgical implantation of osmotic pumps containing beta-estradiol or progesterone into non-pregnant female rabbits resulted in elevated plasma levels of these steroids after 4-5 days at which point the lung levels of FMO B were induced by 35 and 82% for beta-estradiol and progesterone, respectively. The responsiveness of FMO B in lung to both progesterone and glucocorticoids and in kidney to only the latter is consistent with the developmental pattern of both maternal FMO B and peak plasma levels of these steroids during pregnancy.

Aldosterone↗
PubMed1993

20-Hydroxyeicosatetraenoic acid is an endogenous vasoconstrictor of canine renal arcuate arteries.

Recent studies have indicated that renal arteries can produce 20-hydroxyeicosatetraenoic acid (20-HETE) and suggest the potential involvement of a P450 metabolite of arachidonic acid in the myogenic activation of canine renal arteries. In the present study, the effects of 20-HETE on isolated canine renal arcuate arteries were studied. Administration of 20-HETE to the bath or the lumen at concentrations of 0.01-1 microM produced a graded reduction in the diameter of these vessels. In contrast, 19(R)-HETE was a vasodilator, whereas 19(S)-HETE was relatively inactive. The vasoconstrictor response to 20-HETE was not altered by the cyclooxygenase inhibitor indomethacin, endoperoxide/thromboxane receptor antagonist SQ29548, or combined blockade of the cyclooxygenase, lipoxygenase, and P450 pathways using indomethacin, baicalein, and 7-ethoxyresorufin. The response to 20-HETE was associated with depolarization and a sustained increase in the intracellular calcium concentration in renal vascular smooth muscle cells. Patch-clamp studies indicated that 20-HETE significantly reduced mean open time, the open-state probability, and the frequency of opening of a 117-pS K+ channel recorded from renal vascular smooth muscle cells in the cell-attached mode. Microsomes prepared from the renal cortex of dogs produced 20-HETE and 20-carboxyarachidonic acid when incubated with [14C]arachidonic acid. These results indicate that 20-HETE is an endogenous constrictor of canine renal arcuate arteries. The vasoconstrictor response to 20-HETE resembles the myogenic activation of these vessels after elevations in transmural pressure and suggests a potential role for this substance in the regulation of renal vascular tone.

Animals↗
PubMed1993

Strategies for improving maintenance of weight loss. Toward a continuous care model of obesity management.

The management of obesity represents an important objective in the care of many NIDDM patients. In recent years, progress has been made in increasing initial weight reductions, but poor long-term maintenance of weight loss remains a vital clinical concern. This article reviews the challenge of weight-loss maintenance and recommends the adoption of a continuous care model of obesity management. Strategies to improve the long-term maintenance of weight loss are described, and empirical tests of their effectiveness are reviewed. Collectively, the findings suggest that, after treatment for obesity, multifaceted programs comprised of continued professional contact, skills training, social support, and exercise, can enhance the long-term maintenance of weight loss.

Continuity of Patient Care↗
PubMed1993

Anger and sports participation.

This study investigated differences between 19 varsity and 20 intramural male football players in trait anger, anger expression, and sports orientation. While varsity athletes reported comparable levels of trait anger, they described significantly less internalized (anger in) and externalized anger (anger out) than intramural athletes. Also, the varsity athletes reported significantly less anger control. Significant differences were also found for competitiveness and goal orientation, but not win orientation, such that the varsity athletes were more competitive and goal-oriented than the intramural athletes. The implications of these findings are discussed in terms of several alternative hypotheses.

Adolescent↗
PubMed1993

A longitudinal study of intralimb coordination in the first year of independent walking: a dynamical systems analysis.

Using a dynamical systems approach, we examined the development of intralimb coordination over the first year of independent walking. The segmental motion of the thigh and shank and their intralimb coordinative relationship were modeled dynamically as a system of coupled limit cycles. To test the predictions of such a dynamic model in the development of intralimb coordination, 3 infants were filmed from the onset of their first upright independent walking steps, and weekly for the next month and then every month thereafter until they had been walking for 1 year. 3 adults also were included for comparison. Analysis of the kinematic data derived from the film revealed that at the emergence of walking, the limit cycle system displayed instability in several aspects. With walking experience, however, the limit cycle regime became more attractive, such that by 3 months of walking the infants appeared to have found an adult-like stable coordinative relationship between the 2 segments. This pattern of instability followed by stability is consistent with the transitional behaviors of dynamical systems. Several potential control parameters are discussed that might contribute to the stabilization of the walking behavior. The utility of employing a dynamical systems approach for understanding developing behavioral systems is discussed.

Female↗
PubMed1992

Cloned, expressed rat cerebellar nitric oxide synthase contains stoichiometric amounts of heme, which binds carbon monoxide.

The endogenous formation of nitric oxide (NO) has become an area of intense interest as evidence for its biological functions has been obtained in three distinct tissues: circulating macrophages, in which it exerts cytotoxic effects; blood vessels, in which it has been identified as endothelium-derived relaxing factor; and neuronal cells, in which it functions as a neurotransmitter. The formation of NO in brain extracts has been shown to be catalyzed by an enzyme, termed NO synthase, which generates the NO responsible for stimulation of cGMP formation, the highest levels of which occur in the cerebellum. NO synthase catalyzes the formation of citrulline from arginine with the coincident production of NO and has been shown to be a flavoprotein, containing 1 mol each of FAD and FMN, tetrahydrobiopterin, and iron. It is also reported to contain an alpha-helical, calmodulin-binding consensus sequence consistent with its stimulation by calmodulin in the presence of Ca2+. The formation of NO requires incorporation of one of the atoms of molecular oxygen into one of the guanidinium nitrogen atoms of arginine with the coincident formation of citrulline. This communication reports that rat cerebellar NO synthase, cloned and stably expressed in human kidney 293 cells, contains heme in amounts stoichiometric with the flavins FAD and FMN as evidenced by the appearance of a pyridine hemochrome and a reduced CO difference spectrum with an absorbance maximum at approximately 445 nm. The finding of a CO-binding heme moiety explains the presence of iron in the enzyme and suggests a role for prosthetic heme as an oxygenase reaction center. This report also presents evidence for incorporation of delta-[14C]aminolevulinate specifically into immunoprecipitable NO synthase in stably transfected human kidney 293 cells but not in nontransfected cells. Simultaneously, K. A. White and M. A. Marletta [(1992) Biochemistry 31, 6627-6631] have demonstrated a CO-binding heme prosthetic group in purified murine macrophage NO synthase and have suggested the identity of these reaction centers in both the constitutive (cerebellar) and inducible (macrophage) forms of NO synthase.

Amino Acid Oxidoreductases↗
PubMed1991

Bilateral renal hemangiopericytoma.

The first reported case of bilateral renal hemangiopericytoma is presented. The 19 previous renal hemangiopericytomas that have been reported in the world literature are reviewed. This lesion has afflicted patients ranging in age from sixteen to sixty-eight years (mean 41 years) with an equal sex distribution. Mortality from hemangiopericytoma of the kidney has been 50 percent, and the primary mode of therapy remains surgical.

Adult↗
PubMed1991

Development of categorical identification of native and non-native bilabial stops: infants, children and adults.

Using an infant speech identification (ISI) procedure, English language environment infants, two- and six-year-old children, and adults were tested for their identification of sounds on a native (voiced/voiceless bilabial stop) and a non-native (prevoiced/voiced bilabial stop) speech continuum. Categorical perception of the two contrasts diverged as a function of age, increasing for the native contrast and decreasing for the non-native between two and six years. In Experiment 2, subjects of the same four ages were tested for their identification of a continuum of harmonic tones varying in pitch. Comparison of the results of Experiment 1 with the essentially continuous perception of this pitch continuum supports the view that the perception of the native contrast becomes more categorical with age, while perception of the non-native contrast becomes less categorical, especially at six years. Experiment 3, in which adults were tested on the three continua with a multi-trial open set procedure, demonstrated that results with the ISI procedure in Experiments 1 and 2 are comparable to results with more traditional methods. The results of the three experiments are discussed in terms of the role of specific linguistic experience in the development of categorical speech perception.

Adult↗
PubMed1991

Effects of 12-HETE on isolated dog renal arcuate arteries.

12-Hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) is one of the major noncyclooxygenase eicosanoids formed in vascular tissue. The vasoactive effects of the cytochrome P-450 product, 12(R)-HETE and the lipoxygenase product, 12(S)-HETE, on isolated, perfused renal arcuate arteries of the dog were investigated using videomicroscopy. R and S refer to stereoconfiguration of the hydroxy group at the twelfth position of this fatty acid structure. Cumulative doses of 12(R)-HETE produced a concentration-dependent vasoconstriction (n = 9) with a threshold dose of 10(-9) M. At a concentration of 3 x 10(-7) M, 12(R)-HETE reduced vascular diameter by 63 +/- 8 microns (from 306 +/- 17 microns), which was 37 +/- 6% of the maximal vasoconstrictor response to norepinephrine (10(-6) M). The effects of 12(R)-HETE were not altered by indomethacin (10(-6) M, n = 8). The vasoconstrictor response was associated with depolarization of vascular smooth muscle from -47 +/- 1 (15 cells) to -32 +/- 1 mV (12 cells). 12(S)-HETE was also a vasoconstrictor (n = 6), but the threshold concentration for vasoconstriction was 10(-8) M. Small renal arteries obtained from ischemic-injured, but not normal, kidneys produced a metabolite when incubated with arachidonic acid, which coeluted with a 12-HETE standard using reverse-phase high-pressure liquid chromatography. The rate of synthesis of this 12-HETE-like metabolite by renal arteries obtained from ischemic-injured and normal kidneys averaged 1.2 +/- 0.4 (n = 9) and 0.1 +/- 0.1 (n = 7) pmol.h-1.mg tissue-1, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1991

Inhibitors of cytochrome P-450 attenuate the myogenic response of dog renal arcuate arteries.

The role of cytochrome P-450 in the myogenic response of isolated, perfused renal arcuate arteries of dogs to elevations in transmural pressure was examined. The phospholipase A2 inhibitor oleyloxyethylphosphorylcholine (1 and 10 microM) inhibited the greater than threefold increase in active wall tension in these arteries after an elevation in perfusion pressure from 80 to 160 mm Hg. Inhibition of cyclooxygenase activity with indomethacin (1 or 10 microM) had no effect on this response. The cytochrome P-450 inhibitors ketoconazole (10 and 100 microM) and beta-diethyl-aminoethyldiphenylpropylacetate (SKF 525A, 10 and 100 microM) also inhibited the myogenic response. At a pressure of 160 mm Hg, SKF 525A (10 microM) and ketoconazole (100 microM) reduced active wall tension in renal arteries by approximately 70%. Partial inhibition of the myogenic response was obtained after perfusion of the vessels with mechanism-based inhibitors of P-450, 1-aminobenzotriazole (75 microM) and 12-hydroxy-16-heptadecynoic acid (20 microM). The thromboxane receptor antagonist SQ 29,548 (1 or 10 microM) had no effect on the pressure-induced increase in active wall tension in renal arteries. Arachidonic acid (50 microM) constricted isolated perfused renal arteries and potentiated the myogenic response in the presence of indomethacin. This response was completely reversed by ketoconazole (100 microM) or SKF 525A (100 microM). Microsomes (1 mg/ml) prepared from small renal arteries (200-500 microns) and incubated with [1-14C]arachidonic acid (0.5 mu Ci, 50 microM) produced a metabolite that coeluted with 20-hydroxyeicosatetraenoic acid (20-HETE) during reversed-phase high-performance liquid chromatography. The formation of this product was inhibited by both ketoconazole and SKF 525A at concentrations of 10 and 100 microM. These results are consistent with the involvement of the vasoconstrictor 20-HETE and other cytochrome P-450 metabolites of endogenous fatty acids in the myogenic response.

Animals↗
PubMed1991

Apophyseal fracture of the lumbar spine in adolescence.

A case of adolescent lumbar disc herniation with apophyseal fracture is reported. The patient, a 12-year-old boy, experienced sudden onset of pain during athletic activity. Examination revealed tenderness on palpation of the L-5 and S-1 spinous processes, straightening of the lumbar spine, and exquisite pain on 10 degrees of hyperextension. Straight leg raising, Lasègue's, and bowstring tests were all positive. Magnetic resonance imaging and computed tomography confirmed the diagnosis. Treatment included exercise, ibuprofen administration, and limitation of sports activity. At 23 months, the patient is symptom-free. A literature review of 20 cases with computed tomographic or myelographic diagnosis is presented, and conservative and surgical treatment programs are outlined.

Basketball↗
PubMed1990

Cloning and expression of three rabbit kidney cDNAs encoding lauric acid omega-hydroxylases.

cDNAs encoding three cytochrome P-450 enzymes were cloned from a rabbit kidney cDNA library. These three cDNAs exhibit greater than 90% nucleotide sequence identity across the coding region. This degree of sequence identity is also seen with P450IVA4, an enzyme that catalyzes the omega-hydroxylation of prostaglandins and that is elevated during pregnancy and induced by progesterone in rabbit lung. The 3' untranslated regions of the three cDNAs display very little sequence identity, suggesting that they are the products of distinct genes. The predicted amino acid sequences derived from each cDNA and for P450IVA4 exhibit about 85% identity. Each cDNA was inserted into an expression vector for transient transfection of COS-1 cells. The transfected cells each expressed a protein recognized by antibodies to P450IVA4. Microsomes isolated from the cells transfected with each cDNA efficiently catalyzed the omega-hydroxylation of lauric acid with rates that greatly exceed that catalyzed by microsomes isolated from the host cell line. One of the cDNAs encodes an enzyme that omega-hydroxylates prostaglandin A1; however, the specific activity was 2 orders of magnitude lower than that for lauric acid. Our results indicate that the substrate selectivity of the kidney P-450s encoded by these cDNAs is distinct from that of the lung P450IVA4 and that multiple enzymes comprise P-450 class IVA in the rabbit.

Amino Acid Sequence↗
PubMed1989

Viscoelastic shoe insoles: their use in aerobic dancing.

To determine whether use of viscoelastic insoles would significantly decrease the frequency of musculoskeletal overuse injury in aerobic dancers, 139 high-level aerobic dancers were divided randomly into two groups. The control group received placebo foam insoles and test subjects were fitted with viscoelastic insoles. Subjects used these insoles during dance class for 15 weeks. Injury rates were low in both groups and no statistical difference was found. Pain syndromes were fewer in the group using viscoelastic insoles, but the difference was not statistically significant. About a third of dancers fitted with viscoelastic insoles and a tenth of placebo insert wearers found that the insoles made their shoes too tight to be comfortable. No conclusion can be drawn on whether shock-absorbing insoles decrease injuries from aerobic dancing, but use of viscoelastic insoles may improve comfort and provide pain relief for some high-level aerobic dancers if proper fit is achieved.

Adult↗
PubMed1988

Human interlimb coordination: the first 6 months of independent walking.

The organization of interlimb coordination was studied in newly walking infants with and without support and in infants who had been walking 0.5, 1, 3, and 6 months. Analysis of the temporal and distance phase relationships between the limbs revealed that newly walking infants exhibit interlimb coordination similar to that of mature walkers. The coupling between the limbs, however, is loosely constrained, approaching adult-like consistency after 3 months of walking. Providing support for the newly walking infant afforded a more consistent, mature coordination suggesting that neuromuscular development is in advance of its ability to function in the physical world.

Child Development↗