Prevalence of coronary artery disease in patients with symptomatic peripheral vascular disease.

BACKGROUND: Prevalence of coronary artery disease has been reported to be quite high in patients with peripheral vascular disease in western literature. Therefore, it is important to study the coronary anatomy in patients with symptomatic peripheral vascular disease. METHODS AND RESULTS: Fifty-three patients presenting with symptoms of peripheral vascular disease underwent peripheral angiography in our institute during the last 2 years. The total number of vessels involved in these patients was 117. Fifteen patients had involvement of the upper limb vessels, 46 patients had involvement of the lower limb vessels and 6 patients had involvement of the carotid/vertebral arteries. Coronary arteriography was done in all the patients. Only 8 (15%) patients were found to have coronary artery disease with involvement of 11 arteries. Eighty-four (72%) peripheral vessels out of the 117 vessels involved showed total occlusion, whereas only 2 (18%) coronary arteries out of 11 vessels involved showed total occlusion. CONCLUSIONS: This study shows that the majority of patients with symptomatic peripheral vascular disease have normal coronaries, the extent of their involvement being low despite severe peripheral vascular disease.

Regulation of cytoplasmic pH in osteoclasts. Contribution of proton pumps and a proton-selective conductance.

Osteoclasts resorb bone by secreting protons into an extracellular resorption zone through vacuolar-type proton pumps located in the ruffled border. The present study was undertaken to evaluate whether proton pumps also contribute to intracellular pH (pHi) regulation. Fluorescence imaging and photometry, and electrophysiological methods were used to characterize the mechanisms of pH regulation in isolated rabbit osteoclasts. The fluorescence of single osteoclasts cultured on glass coverslips and loaded with a pH-sensitive indicator was measured in nominally HCO(3-)-free solutions. When suspended in Na(+)-rich medium, the cells recovered from an acute acid load primarily by means of an amiloride-sensitive Na+/H+ antiporter. However, rapid recovery was also observed in Na(+)-free medium when K+ was used as the substitute. Bafilomycin-sensitive, vacuolar-type pumps were found to contribute marginally to pH regulation and no evidence was found for K+/H+ exchange. In contrast, pHi recovery in high K+ medium was largely attributed to a Zn(2+)-sensitive proton conductive pathway. The properties of this conductance were analyzed by patch-clamping osteoclasts in the whole-cell configuration. Depolarizing pulses induced a slowly developing outward current and a concomitant cytosolic alkalinization. Determination of the reversal potential during ion substitution experiments indicated that the current was due to H+ (equivalent) translocation across the membrane. The H+ current was greatly stimulated by reducing pHi, consistent with a homeostatic role of the conductive pathway during intracellular acidosis. These results suggest that vacuolar-type proton pumps contribute minimally to the recovery of cytoplasmic pH from intracellular acid loads. Instead, the data indicate the presence of a pH- and membrane potential-sensitive H+ conductance in the plasma membrane of osteoclasts. This conductance may contribute to translocation of charges and acid equivalents during bone resorption and/or generation of reactive oxygen intermediates by osteoclasts.

Carbonic anhydrase II mRNA expression in individual osteoclasts under "resorbing" and "nonresorbing" conditions.

Rabbit osteoclasts can be transformed from a nonresorbing state to a resorbing state by transferring them from culture medium at pH 7.5 to one at pH 6.5. We evaluated whether expression of mRNA for carbonic anhydrase (CA-II) could be used as an indicator of the state of activity of individual osteoclasts. A cDNA probe to rabbit carbonic anhydrase II (CA-II) was prepared and used for in situ hybridization analysis of osteoclasts isolated from neonatal rabbit long bones. Quantitation by grain counting revealed heterogeneity within the osteoclast population: osteoclasts with a "compact" (rounded, less spread) morphology expressed higher levels of CA-II mRNA than "spread" osteoclasts with similar numbers of nuclei. When maintained at pH 6.5 for 6 h, the level of CA-II mRNA was increased significantly in osteoclasts of both morphologies compared with those in parallel cultures maintained at pH 7.5. These results were confirmed by quantitating CA-II mRNA using the polymerase chain reaction (PCR). Oligonucleotide primers specific for rabbit CA-II were synthesized and used to amplify CA-II cDNA transcribed from mRNA prepared from single or small numbers (one to eight cells) of osteoclasts that were collected with a micromanipulator. This generated a approximately 510 bp PCR product, corresponding to the predicted size of the CA-II fragment encompassed by the primers. For quantitation, CA-II mRNA levels were compared with the levels of a approximately 900 bp actin fragment that was coamplified in the same reaction mixture or amplified separately in a duplicate sample of the reaction mixture.(ABSTRACT TRUNCATED AT 250 WORDS)

Early inhibition of myointimal proliferation by angiopeptin after balloon catheter injury in the rabbit.

PURPOSE: Coronary artery restenosis after percutaneous transluminal angioplasty occurs in more than 40% of patients. Angiopeptin, a stable synthetic octapeptide analogue of somatostatin, attenuates accelerated coronary artery myointimal thickening in rabbit cardiac allografts and myointimal thickening after arterial injury. In this study the temporal relationship between the angiopeptin treatment schedule and efficacy was explored. The relationship between inhibition of myointimal thickening by angiopeptin and inhibition of vascular cell proliferation was also examined. METHODS: The aorta and the common and external iliac arteries of the rabbit underwent balloon injury. Angiopeptin (2 to 200 micrograms/kg/day) was administered for 1 day before injury and for 1, 5, and 21 days after injury. Morphometric studies were performed to determine measurement of intimal thickening. Inhibition of vascular cell proliferation by angiopeptin was evaluated by tritiated thymidine incorporation into the balloon-injured rabbit aorta. Thymidine was either administered intraperitoneally or added ex vivo to aorta segments of rabbits treated with angiopeptin (2, 20, or 200 micrograms/kg/day) from 1 day before injury until sacrifice 72 hours later. RESULTS: Administration of angiopeptin (2 to 200 micrograms/kg/day) significantly reduced intimal thickening by approximately 50% in all three vessels when evaluated 3 weeks after injury. This inhibitory effect was unrelated to duration of treatment and dose. Treatment initiated at the time of injury was found to be effective, but delaying treatment for 8, 18, or 27 hours abrogated the inhibitory effect of angiopeptin on myointimal thickening. Angiopeptin treatment significantly decreased thymidine-labeled nuclei of smooth muscle cells in vitro. Angiopeptin treatment similarly inhibited thymidine uptake in vitro by balloon-injured aorta segments. CONCLUSION: Angiopeptin significantly inhibits myointimal thickening by inhibiting vascular cell proliferation. Administration of angiopeptin for 2 days is as efficacious as 3 weeks treatment in inhibiting myointimal thickening. Delaying treatment for as little as 8 hours after injury abrogates the inhibitory effects of angiopeptin. This speaks to the importance of early events immediately after vascular tissue injury, suggesting that angiopeptin inhibits the expression of early genes causally related to the vascular injury response and thereby triggering vascular cell proliferation.

Estradiol inhibition of arterial neointimal hyperplasia after balloon injury.

PURPOSE: Estrogens have been shown to protect against cardiovascular disease in postmenopausal women. The mechanisms are unknown. In this study we investigated the effect of estrogen treatment on arterial neointimal proliferation elicited by balloon injury of blood vessels of the rabbit. METHODS: The aorta and the common and external iliac arteries of the rabbit underwent balloon injury. 17-beta-Estradiol cypionate (100 micrograms/kg/day intramuscularly) was administered beginning 1 day before injury and until sacrifice at 22 to 24 days after injury. Angiopeptin (20 micrograms/kg/day) was administered for the same length of time as estrogen to a group of rabbits to use this peptide as a positive control for morphometric analysis. Angiopeptin was also combined with estradiol to determine whether it was possible to further enhance the effect of estrogen. Morphometric studies were performed to determine measurement of intimal thickening. Inhibition of cell proliferation by estrogen was evaluated by incorporation of tritiated thymidine in vitro into the balloon injured rabbit aorta 72 hours after balloon injury. RESULTS: Treatment of male rabbits with estradiol significantly (p < 0.01) decreased the neointimal thickening of these vessels by 50% to 70%. The somatostatin analog, angiopeptin, was similarly effective in the same circumstances. Estradiol failed to further inhibit neointimal thickening when combined with angiopeptin. Treatment with estradiol for 3 days inhibited both thymidine incorporation (p < 0.01) and DNA content in injured vessels. CONCLUSION: Estradiol treatment of rabbits undergoing balloon injury of the aorta and iliac arteries, significantly inhibits the myointimal thickening. This effect of estrogen is mediated by inhibition of vascular smooth muscle cell proliferation.

Biosynthesis, catabolism, and biological properties of HPETEs, hydroperoxide derivatives of arachidonic acid.

The oxygenation of arachidonic acid by lipoxygenases results in the formation of HPETEs (hydroperoxyeicosatetraenoic acids), the first products of the LOX pathway. These compounds are short lived and are catabolised into various families of more stable compounds of which the HETEs, hepoxilins, lipoxins and leukotrienes have been identified so far. The development of new techniques have helped to identify and understand the structures of various HPETEs and only recently the biological effects of HPETEs and their various catabolites are being unraveled. Although lipoxygenases are ubiquitous, not all tissues possess the same spectrum of lipoxygenase enzymes. Hence different HPETEs can be formed in different tissues. Recent studies have revealed that HPETEs or products derived from them possess a diversity of important biological properties including the regulation of electrolyte flux and eicosanoid and corticosterone syntheses, release of histamine, regulation of oocyte maturation and release of various reproductive hormones. HPETEs appear to be involved in some pathological conditions viz, skin psoriasis, Clarkson's disease, nerve injury and spinal cord ischemia. These novel eicosanoids are associated with the release of insulin as well as renin. Recently HPETEs have been suggested to act as second messengers in the Aplysia sensory neurons and its catabolite, hepoxilin, has been demonstrated to have effects on mammalian hippocampal neurons. The purpose of this review is to provide a brief summary of the formation of the HPETEs and the various families of compounds derived from them as well as the various types of biological activities for these products described so far.

Polyamines and cell proliferation in the sea star Pycnopodia helianthoides.

1. Diamines (putrescine and cadaverine) and polyamines (spermidine and spermine) were extracted from tissues of the sea star Pycnopodia helianthoides, separated and quantitated using reversed-phase high-performance liquid chromatography (RP-HPLC). Simultaneous measurements of levels of protein and DNA and rates of incorporation of 14C-thymidine were carried out. 2. The most abundant polyamine in tissues was spermidine (0.3873-2.5282 nmol/mg tissue) followed by spermine (0.103-1.5517 nmol/mg tissue), putrescine (0.2096-0.5322 nmol/mg tissue) and cadaverine (0.022-0.6064 nmol/mg tissue). 3. An unknown molecule with derivatization and elution behaviour similar to that of polyamine standards was detected in all tissues. 4. Protein levels ranged from 20.47 mg/g tissue in the body wall to 48.44 mg/g tissue in the pyloric caecum. 5. DNA levels were lowest in the ovary (0.25 mg/g tissue) and highest in the testis (5.62 mg/g tissue). 6. Incorporation of 14C-thymidine was highest in the testis. Testicular tissue had the highest spermidine/spermine ratio (5.4). A significant correlation between the spermidine/spermine ratio and 14C-thymidine incorporation (expressed either as DPM/g tissue or DPM/mg protein) suggests that polyamines are implicated in the regulation of cell proliferation in the sea star P. helianthoides.

Alanine and aspartate aminotransferases in normal and denervated skeletal muscle.

Activities of alanine and aspartate aminotransferases are maintained during the first 5 weeks of growth, but decrease subsequently in normal chick gastrocnemii. In sciatectomized muscles, a 5-fold elevation in these enzymes reveals increased utilization of amino acids as a compensatory metabolic support during denervation atrophy.

Macromolecular biosynthesis in Mycobacterium smegmatis ATCC 607 in the presence of antibodies to mannophosphoinositides.

The biosynthesis of DNA, proteins, RNA and phospholipids in Mycobacterium smegmatis ATCC 607 was investigated by studying the incorporation of radiolabelled components in the presence of antiserum to mannophosphoinositides. The antiserum had an inhibitory effect on the rate of synthesis of these macromolecules. However, the inhibition was greater when antibody was present together with complement.