Identification of albumin precursor protein, Phi AP3, and alpha-smooth muscle actin as novel components of redox sensing machinery in vascular smooth muscle cells.

Aerobic organisms are continually subjected to environmental stressors that compromise redox homeostasis and induce cellular injury. In vascular smooth muscle cells (vSMCs), the activation/repression of redox-regulated genes after environmental stress often involves protein binding to cis-acting antioxidant response elements (AREs). The present study was conducted to identify proteins that participate in redox-regulated protein binding to human c-Ha-ras and mouse glutathione S-transferase A1 AREs in vSMCs after oxidant injury. Challenge of vSMCs with 0.3 or 3 microM hydrogen peroxide, 3-methylcholanthrene, benzo[a]pyrene-7,8-diol, 3-hydroxy benzo[a]pyrene, and benzo[a]pyrene-3,6-quinone induced concentration-related increases in ARE protein binding. The profiles of ARE complex assembly were comparable, but exhibited chemical specificity. Pretreatment with 0.5 mM N-acetylcysteine inhibited activation of ARE protein binding in hydrogen peroxide-treated cells. Preparative electrophoretic mobility shift assays coupled to Western analysis identified NF-E2-related proteins 1 and 2 and JunD in complexes assembled on AREs. Polyethylenimine affinity and sequence-specific serial immobilized DNA affinity chromatography followed by N-terminal sequencing identified albumin precursor protein, phi AP3, and alpha-smooth muscle actin as members of the ARE signaling pathway. Sequence analysis of albumin protein revealed homology to the redox-regulated transcription factors Bach1 and 2, as well as cytoskeletal and molecular motor proteins. These results implicate albumin precursor protein, phi AP3, and alpha-smooth muscle actin as participants in redox sensing in vSMCs, and suggest that protein complex assembly involves interactions between leucine zipper and zinc finger transcription factors with cytoskeletal proteins.

Impact of cellular metabolism on the biological effects of benzo[a]pyrene and related hydrocarbons.

Polycyclic aromatic hydrocarbons are ubiquitous contaminants in the environment. Benzo[a]pyrene (BaP), a prototypical member of this class of chemicals, has been extensively studied for its toxic effects in laboratory animals and human populations. BaP toxicity is often mediated by oxidative metabolism to reactive intermediates that interact with macromolecules leading to alterations in target cell structure and function. More recent evidence suggests that disruption of cellular signaling pathways involved in the regulation of growth and differentiation contribute significantly to the toxicity of BaP and its metabolites. This review summarizes recent advances in our understanding of biological mechanisms of BaP toxicity at the molecular level, and the role of metabolic intermediates in carcinogenesis, atherogenesis, and teratogenesis.

Profiles of antioxidant/electrophile response element (ARE/EpRE) nuclear protein binding and c-Ha-ras transactivation in vascular smooth muscle cells treated with oxidative metabolites of benzo[a]pyrene.

Activation of nuclear protein binding to the antioxidant/electrophile response element (ARE/EpRE) by benzo[a]pyrene (BaP) in vascular smooth muscle cells (vSMCs) is associated with transcriptional deregulation of c-Ha-ras. This response may be mediated by oxidative intermediates of BaP generated during the course of cellular metabolism. To test this hypothesis, the profile of ARE/EpRE protein binding and transactivation elicited by BaP was compared with that of 3-hydroxy BaP (3-OH BaP) (0.03 to 3.0 microM), BaP 7,8-dihydrodiol (BaP 7,8-diol) (0.03 to 3.0 microM), BaP 3,6-quinone (BaP 3,6-Q) (0.0003 to 3.0 microM), and H(2)O(2) (25 to 100 microM). Specific protein binding to the consensus c-Ha-ras ARE/EpRE was observed in vSMCs treated with all BaP metabolites at concentrations considerably lower than those required for the parent compound. H(2)O(2), a by-product of BaP 3,6-Q redox cycling, also increased binding to the ARE/EpRE. Treatment of vSMCs with oxidative BaP metabolites or H(2)O(2) transactivated the c-Ha-ras promoter in all instances, but the response was consistently half of the maximal induction elicited by BaP. Similar proteins cross-linked specifically to the consensus c-Ha-ras ARE/EpRE sequence in cells treated with BaP or its oxidative intermediates. The protein binding profile in the c-Ha-ras promoter was similar to that in the NADPH:quinone reductase gene (NQO(1)) and the glutathione S-transferase Ya gene (GSTYa) promoters, but the relative abundance of individual complexes was promoter-specific. We conclude that oxidative intermediates of BaP mediate activation of nuclear protein binding to ARE/EpRE and contribute to transcriptional de-regulation of c-Ha-ras in vSMCs.

Activation of nuclear protein binding to the antioxidant/electrophile response element in vascular smooth muscle cells by benzo(a)pyrene.

This laboratory has previously shown that binding of nuclear proteins to the antioxidant/electrophile response element (ARE/EpRE) participates in deregulation of vascular gene expression by benzo(a)pyrene (BaP), a suspected atherogen. In the present study, oligonucleotides representing ARE/EpREs within the c-Ha-ras and glutathione-S-transferase (GST-Ya) promoters were employed to evaluate the role of flanking sequences in stabilizing protein:DNA interactions in BaP-treated vascular smooth muscle cells (vSMCs). We also wanted to define promoter-specific patterns of protein recognition to ARE/EpREs in this cell system. In electrophoretic mobility shift assays (EMSA), optimal protein binding to a human Ha-ras ARE/EpRE variant sequence fitted to match the extended mouse(m) GST-Ya ARE/EpRE core (5'-TMAnnRTGAYnnnGCR-3') was dependent on 5' nucleic acid sequence. Using immobilized DNA affinity chromatography (IDAC), we identified four nuclear proteins of M(r) 62, 60, 50, and 30 kDa that associated specifically with the mGSTYa ARE/EpRE. Photo crosslinking to a BrdU-substituted hHa-ras or mGST ARE/EpRE probe identified specific proteins of M(r) 80, 60, 55, 25, 23 kDa or 80, 60, 55, 27, 25, 23 kDa, respectively. Protein:DNA complexes detected using IDAC eluate overlapped with those observed in crude nuclear extracts. Chemical treatments known to modulate ARE/EpRE protein binding in vSMCs did not alter overall protein:DNA affinity and/or sequence recognition to either hHa-ras or mGST-Ya elements. We conclude that nucleotide sequences 5' to the core ARE/EpRE influence specific binding of nuclear proteins and that multiple proteins bind to ARE/EpREs in a promoter-specific manner in vSMCs.

Success rate analysis of Navy SERGRAD flight training.

SERGRADS are selectively retained graduates of Naval Flight Training who become flight instructors during their first tour as Naval Aviators. A perception exists that SERGRADS have more difficulties than newly designated pilots during subsequent training in the Fleet Replacement Squadrons (FRS). A preliminary inquiry at the F/A-18 FRS revealed that former SERGRADS appear to experience a higher rate of failure on their initial carrier qualification attempt during FRS training. This study examined the SERGRAD experience and compared their performance to trainees from other sources. Although significant differences were found between groups for Fleet Replacement Squadron flight grades and Training Command composite grades, no significant differences were found between the groups for Training Command carrier qualification grades or Fleet Replacement Squadron carrier qualification. Thus, the data do not support the hypothesis that SERGRADS have more problems or are poorer aviators than F/A-18 students from other sources.

Evaluation of various nitrogen supplements in starter diets for growing Holstein steers and their effects on ruminal bacterial fermentation in continuous culture.

Concurrent in vivo and in vitro studies were conducted to evaluate urea (U), soybean meal (SBM), ground soybeans (RAW), extruded soybeans (ES) or extruded soybeans plus urea (ES + U) as primary supplemental N sources in starter diets for Holstein steers. Three groups of 48 Holstein steers each were fed five different starter diets to 181 kg BW in three experimental periods over 2 yr. Average daily gains were similar (P greater than .05) for steers fed ES + U (1.12 kg), ES (1.08 kg) and SBM (1.09 kg) but lower (P less than .05) for those fed U (1.00 kg) or RAW (.97 kg) diets. Feed/gain was similar (P greater than .05) for ES-fed steers vs those fed other diets except U. From 181 to 477 kg, all steers were fed the same diet. Steers fed the RAW starter diet had the lowest (P less than .05) ADG for the entire period. The starter diets were used as substrates for ruminal microbial metabolism in eight dual-flow continuous culture fermenters. True OM digestion was higher and NDF and ADF digestion was lower (P less than .05) for the ES + U diet than for the ES diet. Dietary protein degradation was lowest (P less than .05) for the ES diet (64.4%). Total bacterial N flow was higher (P less than .05) with the ES + U, SBM and U diets than with the ES diet. Lysine flow was higher (P less than .05) for the ES + U diet than for all other diets except ES. Results of these experiments indicate that ES as a protected ruminal escape N source with or without added urea did not improve steer performance above that obtained from SBM in starter diets.

The cardiac manifestations of acquired immune deficiency syndrome.

This article reviews the cardiac manifestations of acquired immune deficiency syndromes (AIDS) as reported in the literature. These abnormalities were examined with regard to incidence, clinical presentation, and possible etiologies. Review of the literature revealed that cardiac abnormalities occur frequently in persons with AIDS; involve the heart valves, myocardium, and pericardium; and, in many cases, may not arouse clinical suspicion. Because the cardiac sequelae that may occur during the course of the disease can be life-threatening, the nurse must be alert to the possibility of cardiac involvement and initiate prompt intervention when cardiac complications occur.

Antiplatelet therapy for ischemic heart disease in the elderly.

A vast experience has developed defining an important role for blood platelets in the etiology of atherosclerotic coronary disease. Antiplatelet drugs such as aspirin have major therapeutic applications in various ischemic heart disease syndromes. Areas of proven efficacy for antiplatelet therapy include maintenance of saphenous vein graft patency and prevention of myocardial infarction and death in patients with unstable angina and in survivors of myocardial infarction. Other less-defined applications include primary prevention and use during acute myocardial infarction with and without thrombolysis. The issues of antiplatelet efficacy in women, drug doses, and risk of toxicity need further clarification.

Platelets and antiplatelet therapy in ischemic heart disease.

Blood platelets have been shown to play an important role not only in thrombosis, but also in the pathogenesis of coronary artery disease and its complications. Drugs that affect platelets have been shown to reduce mortality in survivors of acute myocardial infarction, to reduce the risk of myocardial infarction in patients with unstable angina, and to preserve the potency of saphenous venous grafts used to bypass obstructed coronary arteries. The drugs may also play a role in the primary prevention of arteriosclerosis and in preventing thrombotic complications following coronary angioplasty.

Effects of oral doses of corn oil and other factors on preweaning survival and growth of piglets.

Piglets were provided supplemental energy as oral doses of corn oil to find whether such treatment might improve survival or weight gains. A total of 1,840 piglets from 182 litters was used. Piglets were allotted to dosed and control treatments within litter on the basis of body weight. Dosed piglets received a total of 8 ml of corn oil in four doses during the first 48 h postpartum. The survival rate to weaning was high and not affected (P greater than .10) by the corn oil doses. Deaths of piglets low in birth weight were delayed by the fat doses, suggesting some utilization. The fat doses did not affect piglet weight gain. Factors affecting preweaning survival and growth of piglets were also evaluated using data from the same piglets. Specific factors investigated were sex, parity of dam, birth weight, litter size and the mean and standard deviation of piglet birth weights within litters. Male piglets were heavier at birth than females, but there was no sex effect on weight at 7 d or at weaning. Small males were less likely to survive to 3 d than were small females. Litters from primiparous sows were smaller at birth and weaning, but had lower percentages of stillbirths and of preweaning deaths than did those from multiparous sows. Piglets from primiparous sows were more uniform in birth weight. Piglets that were heavier at birth grew faster and were more likely to survive. Litter size affected growth but not survival of piglets. A heavier average birth weight in the litter reduced slightly the growth rate of individual piglets. An increase in variability of piglet birth weight within a litter was associated with an increase of the percentages of stillbirths and of preweaning mortality. A few litters accounted for a major portion of the mortality. Much of the variation in piglet mortality and growth was not explained by factors considered.

Correlated response of udder dimensions to selection for milk yield in Holsteins.

Response to selection for milk yield in Holsteins was examined by a controlled selection experiment initiated in 1964. Foundation cows were paired by sire and divided randomly into two breeding groups, selection and control. Selection group was mated to four sires each year highest for Predicted Difference milk. Twenty bulls selected in 1964 as near breed average for milk sired all control cows. Milk yield was recorded for each lactation, and height of udder from ground, distances between teats, and perimeter and area bound by the four teats were measured at 30 to 75 days postpartum on 153 selection and 202 control cows over 14 yr. Most estimates of repeatability and heritability of udder dimensions before and after milking were .45 or larger. Correlations with milk yield were negative for udder height and positive for other udder measurements. Selection cows exceeded controls for lactational milk yield. Daughters of high milk bulls had greater distances between teats, greater perimeters, and larger areas of udder floor. Selection cows did not differ from controls in udder height for first parity but were below controls in udder height and had udders that collapsed more for all parities.

Severe hyperkalemia during piroxicam therapy.

Severe hyperkalemia developed in an 85-year-old man after he had been receiving piroxicam treatment for several months. At admission his serum potassium level was 9.3 mEq/L; total CO2 level, 11 mmole/L; chloride level, 122 mEq/L; serum urea nitrogen level, 54 mg/dL; and creatinine level, 2.5 mg/dL. Hyperkalemia resolved after withdrawal of the drug and polystyrene sodium sulfonate therapy and the nonanion gap acidosis subsided concomitantly. His serum urea nitrogen and creatinine levels remained unchanged. He had abnormally low plasma renin activity, which gradually returned to normal, and aldosterone concentration, which remained low. The nonsteroidal drug may have impaired renin secretion, adrenal responsiveness to angiotensin, or the action of aldosterone on the renal tubule.

Evaluation of high protein oats as a replacement for corn in diets fed to growing-finishing swine.

In a series of five trials, 680 crossbred pigs were fed isolysine diets in which high protein (13.8 to 16.0%) oats constituted 0, 20, 40 or 60% of the cereal portion. There was a linear depression in average daily gain (P less than .001) and average daily feed intake (P less than .005) with increasing level of oats, but feed/gain was unaffected (P less than .05) during the growing period from 22 to 46 kg. There were no significant differences in average daily gain during the finishing period from 46 to 102 kg; however, feed/gain increased linearly (P less than .01) with increasing level of oats in the diet. Increases in the percentage of oats in the cereal portion of the diet resulted in a linear (P less than .01) decrease in average daily gain and an increase (P less than .01) in feed/gain for the entire feeding period. We conclude that high protein-high lysine oats of the quality used in these trials can be included at up to 20% of the cereal portion of the diet without greatly affecting performance.

Emergency medical services: teamwork to fight sudden death.

It is estimated that upwards of 20% of accidental highway and prehospital coronary deaths could be averted by the availability of prompt, effective prehospital care; this implies that approximately 35,000 deaths from myocardial infarctions and 12,000 deaths from vehicular trauma are preventable. So, statistically, are 13,000 deaths from cerebrovascular accidents, poisonings, drowning, and other accidents. There are nearly 12 million nonfatal injuries each year, the suffering, cost and disabilities of which are incalculable. It is clear that these statistics demand some type of unified multi-professional response. Although not perfect, Emergency Medical Services is clearly the most effective available means of reversing the present rates of sudden death and disability.

Health care requirements of dairy cattle. I. Response to milk yield selection.

To measure direct response to single trait selection for milk yield and correlated response in health problems, two homologous base populations were formed by pairing 66 Holstein females by sire. Base populations and descendants were managed identically except for selection by milk yield. One base and lineage were mated with sires with highest predicted difference milk (selection group); other base and lineage were mated with average sires in 1964 (control group). Milk yield, supplemental labor, and veterinary and semen expense were recorded specific to each cow. Over 9 yr, 130 selection and 163 control cows were observed. Selection cows yielded more milk but with increase in labor and expense for health care. Estimates of labor for the selection group were greater for mammary, locomotion, and digestion categories and for expense in mammary, respiration, first insemination, and later insemination categories. Labor and expense for reproduction did not differ for genetic groups. Differences between groups in annual estimated labor and expense totaled $27.00. Extra income over feed cost more than compensated for greater health care and semen cost; however, the magnitude of the latter indicates a need for them to be considered when economically evaluating breeding programs where major emphasis is on milk yield.