Oxidant stress and glioblastoma multiforme risk: serum antioxidants, gamma-glutamyl transpeptidase, and ferritin.

Case-control studies of serum antioxidants are difficult to interpret, because antioxidants may be altered by the disease under study. However, because glioblastoma multiforme (GBM) is a relatively rare disease, a cohort study would require a large sample observed for many years. In the present case-control pilot study (34 cases and 35 controls), we evaluated the association between serum levels of ascorbic acid (AA) and alpha- and gamma-tocopherol (alpha-T and gamma-T) measured before diagnostic surgery. To control for influence of GBM on serum AA, alpha-T, and gamma-T, we adjusted for oxidant stress indexes (gamma-glutamyl transpeptidase and uric acid) and an acute-phase response index (serum ferritin). When adjusted, AA is inversely related to GBM (p for trend = 0.007). In addition, AA interacts with alpha-T to further reduce GBM risk (test for interaction, p = 0.04). gamma-T is not associated with GBM (p = 0.71). However, gamma-glutamyl transpeptidase (p = 0.004), coenzyme Q (p = 0.01), and ferritin (p = 0.009) are positively and uric acid (p = 0.000) is negatively related to GBM. We conclude that 1) AA and alpha-T are jointly related to GBM after adjustment for GBM-produced oxidant stress and 2) there is a strong association between the presence of GBM and oxidant stress.

Hypotheses concerning roles of dietary energy, cured meat, and serum tocopherols in adult glioma development.

We conducted a pilot study to determine whether: (1) high levels of energy intake increase glioma risk; (2) the previously observed relationship between cured meat consumption and glioma risk can be attributed to confounding by energy intake, and (3) alpha-tocopherol modifies caloric intake and gamma-tocopherol modifies cured meat consumption. We identified 40 age-sex-race matched glioma sets, and obtained serum vitamin C and alpha- and gamma-tocopherol levels for 23 of these sets. Glioma risk increased with quartile of total dietary energy adjusted for fat, protein, and nitrite-containing meat consumption (odds ratio = 1.0, 2.7, 5.8, 8.2; p value for trend test = 0.02). Although positive associations between individual cured meats and glioma risk decreased when adjusted for caloric intake, the small sample size makes it difficult to interpret the results. Serum alpha-tocopherol appeared to modify the effect of calories and serum gamma-tocopherol may modify the effect of cured meat on glioma risk. While the observed interaction is predicted by experimental research, our findings are based on small numbers. Larger studies are needed to further evaluate our preliminary findings.

Cytotoxicity of tocopherols and their quinones in drug-sensitive and multidrug-resistant leukemia cells.

Cytotoxicities of tocopherols (alpha-T, gamma-T, delta-t), their para (alpha-TQ, gamma-TQ, delta-TQ)- and ortho (Tocored)-quinone oxidation products, the synthetic quinone analog of gamma-TQ containing a methyl group substituted for the phytyl side-chain (TMCQ) and the synthetic quinone analog of Tocored containing a methyl group substituted for the phytyl side-chain (PR) were measured in acute lymphoblastic leukemia cell lines that are drug-sensitive (CEM) and multidrug-resistant (CEM/VLB100). Among tocopherols, only delta-T exhibited cytotoxicity. Among para quinones, alpha-TQ showed no cytotoxicity, while gamma-TQ and delta-TQ were highly cytotoxic in both CEM and CEM/VLB100 cell lines (LD50 < 10 muM). delta-TQ and gamma-TQ were more cytotoxic than the widely studied chemotherapeutic agent doxorubicin, which also showed selective cytotoxicity to CEM cells. The orthoquinone Tocored was less cytotoxic than doxorubicin in drug-sensitive cells but more cytotoxic than doxorubicin in multidrug-resistant cells. Cytotoxicity was not a function of the phytyl side-chain since both TMCQ and PR were cytotoxic in leukemia cells. Cytotoxic para and ortho quinones were electrophiles that formed adducts with nucleophilic thiol groups in glutathione and 2-mercaptoethanol. Cytotoxicity was enhanced when the glutathione pool was depleted by preincubation with buthionine-[S,R]-sulfoximine, but cytotoxicity was diminished by the addition of N-acetylcysteine to cultures. alpha-T also diminished the cytotoxicity of para- and orthoquinones. Buthionine-[S,R]-sulfoximine did not block the inhibitory effect of either N-acetylcysteine or alpha-T, showing that these agents did not act solely by maintaining the glutathione pool as an essential antioxidant system. In conclusion, tocopherylquinones represent a new class of alkylating electrophilic quinones that function as highly cytotoxic agents and escape multidrug resistance in acute lymphoblastic leukemia cell lines.

Closely related genes encode developmental and tissue isoforms of porcine cytochrome P450 aromatase.

We examined the chromosomal basis for the synthesis of tissue (ovary, endometrium/placenta, and peri-implantation blastocyst) isoforms of cytochrome P450 aromatase in the pig. DNA fragments derived from three distinct porcine aromatase chromosomal genes were cloned and characterized. The porcine type III aromatase gene encoding the blastocyst aromatase isoform was found to consist of nine coding exons and two mutually exclusive, 5' untranslated exons (designated E1A and E1B), collectively spanning 30 kb or more. The porcine type II aromatase gene, encoding the endometrial/placental aromatase isoform, was identified by cloning of a genomic DNA fragment spanning the corresponding exons 7, 8, and 9. The DNA inserts of two other phage clones encompassed exons 2, 3, and 4 of a third chromosomal gene (type I) encoding the ovarian aromatase isoform. All intron-exon junctions in these genomic fragments were found to be identical in relative positions to those of the single-copy human aromatase gene. Comparisons of cDNA and genomic sequences indicated that nucleotide sequence variation was not uniform across the corresponding exons of these genes and that the corresponding intronic sequences were conserved. The type II and type III aromatase genes were localized to the same regional location (q16-17) on swine chromosome 1, which is homologous to the human chromosome 15 region (q21.1) in which the human aromatase gene resides. Results demonstrate that the three aromatase genes characterized in the present study appear to be similar in their overall structural organization and most likely are clustered, which could have resulted from at least two independent gene duplication events. The presence of multiple aromatase genes constitutes a newly described mechanism by which aromatase enzyme biosynthesis and functional activity can be regulated in a tissue and temporal fashion and serves to highlight further the complexity of aromatase gene expression in mammals. Moreover, the presence of a unique aromatase gene that is highly expressed in pig blastocysts may constitute a paradigm for other mammals (e.g., equids, rabbit, hamster) whose peri-implantation blastocysts are estrogenic.

Physical mapping of SOD1 to bovine chromosome 1.

Superoxide dismutase 1 (SOD1) was mapped to cattle chromosome 1q12 --> q14 by in situ methods. Both traditional in situ hybridization using tritium and a new technique, direct in-situ single copy PCR (DISC-PCR), were used in two separate laboratories. Both human and bovine SOD1 clones were tritium labeled for radioactive in situ hybridization. A primer pair based on the bovine SOD1 gene (Barendse et al., 1994b) was used for the DISC-PCR procedure. The map location of SOD1 is close to collagen 6A1. SOD1 is a potentially important type 1 anchor locus in the region where the gene for horns in cattle was recently mapped (Georges et al., 1993; Schmutz et al., 1995).

Directed integration of the physical and genetic linkage maps of swine chromosome 7 reveals that the SLA spans the centromere.

The first integrated physical and genetic linkage map encompassing the entire swine chromosome 7 (SSC7) reveals that the porcine MHC (SLA) spans the centromere. A SLA class II antigen gene lies on the q arm, whereas class I and III genes lie on the p arm, suggesting that the presence of a centromere within the SLA does not preclude a functional complex. The SLA appears smaller than other mammalian MHC, as the genetic distance across two class I, three class II, and three class III SLA gene markers is only 1.1 cM. There are significant variations in recombination rates as a function of position along the chromosome, and the SLA lies in the region with the lowest rate. Furthermore, the directed integration approach used in this study was more efficient than previous efforts that emphasized the screening of large insert libraries for random microsatellites.

Comparative analyses of peritoneal fluid from calves and adult cattle.

Reference values for hematologic variables change with increasing age in cattle. Therefore, the purpose of the study reported here was to describe the peritoneal fluid constituents of clinically normal young calves, and to compare cellular concentration and distribution in blood and peritoneal fluid of young calves with those of adult cattle. Eight healthy 8-week-old male Holstein calves and 8 healthy 3- to 8-year-old Holstein cows were studied. Peritoneal fluid was collected from calves along the ventral midline, 4-cm cranial to the umbilicus. Abdominocentesis was performed in the region of the lower right flank in adult cattle. Correlation analysis, using the Pearson's correlation coefficient, and regression analysis were performed for blood and peritoneal fluid data from calves. Data from calves were compared with those of cows, using Wilcoxon's rank sum test. A P value < 0.05 was considered significant for all tests. Calves had significantly lower blood eosinophil count (P < 0.003) and plasma protein concentration (P < 0.001) than did cows. Calves had significantly higher peritoneal fluid nucleated cell (P < 0.05) and mononuclear cell (P < 0.05) counts, but lower peritoneal fluid eosinophil cell count (P < 0.003) than did cows. For calves, nucleated cell and lymphocyte cell counts in the blood had a high, positive correlation with those of peritoneal fluid. However, the prediction equation for nucleated cell count accounted for a modest proportion of variability. A prediction equation for peritoneal fluid lymphocyte cell count was established.(ABSTRACT TRUNCATED AT 250 WORDS)

Antioxidant and cytotoxic tocopheryl quinones in normal and cancer cells.

We found previously that [d]-alpha-tocopherol (alpha-T) and [d]-gamma-tocopherol (gamma-T) are lipid antioxidants (thiobarbituric acid test) in model systems containing arachidonic acid (AA), cumene hydroperoxide, and Fe3+ and in smooth muscle cell (SMC) cultures challenged with AA. We now show that [d]-alpha-tocopherylquinone (alpha-TQ), [d]-delta-tocopherylquinone (delta-TQ), and [d]-gamma-tocopherylquinone (gamma-TQ) are antioxidants at low concentrations and prooxidants at high concentrations in the model system. Prooxidant activity is greater with gamma-TQ than either alpha-TQ or delta-TQ. Low concentrations of alpha-TQ, delta-TQ, and gamma-TQ are also antioxidants in SMC cultures challenged with AA. Unlike alpha-TQ, partially substituted gamma-TQ and glutathione (GSH) form a Michael adduct which has been purified and characterized. We found previously that alpha-T, gamma-T, and alpha-TQ are mitogenic in SMC. We now report that both delta-TQ and gamma-TQ but not alpha-TQ show concentration-dependent cytotoxicity (changes in morphology, propidium iodide stain) in SMC cultures. Cytotoxicity is greater with gamma-TQ than delta-TQ. An acute lymphoblastic leukemia (ALL) cell line shows greater chemosensitivity (MTT and Neutral Red assays) to gamma-TQ than to either doxorubicin (DOX) or vinblastine (VLB). An ALL cell line resistant to both DOX and VLB retains the same chemosensitivity to gamma-TQ as the drug-sensitive ALL cell line. ALL cell lines are unaffected by either alpha-TQ or the GSH Michael adduct of gamma-TQ. These data show that partially substituted tocopheryl quinones capable of forming Michael adducts are potential chemotherapeutic agents for multidrug-resistant cancer cells.

Comparison of peritoneal fluid analysis before and after exploratory celiotomy and omentopexy in cattle.

The effect of right paralumbar fossa exploratory celiotomy and omentopexy on peritoneal fluid constituents was studied in 22 adult dairy cows. Six cows were eliminated on the basis of physical examination findings (n = 2), surgical findings (n = 2), or inability to obtain a sufficient volume of peritoneal fluid (n = 2). Sixteen cattle had normal results of CBC and serum biochemical analysis, and a minimum of 1 ml of peritoneal fluid was obtained by abdominocentesis. Abdominocentesis was repeated on days 1, 2, and 6 after surgery. Statistical analysis for repeated measures was performed, using a significance level of P < 0.05. Stage of gestation was evaluated for interaction with time. Mean total nucleated cell count was 3,200 cells/microliters before surgery, was significantly increased 2 days after surgery (16,336 cells/microliters), and continued to increase through day 6 (20,542 cells/microliters). Mean polymorphonuclear cell count was 1,312 cells/microliters before surgery and was significantly higher at 2 (11,043 cells/microliters) and 6 (10,619 cells/microliters) days after surgery. Mean lymphocyte count was 254 cells/microliters before surgery and was significantly increased 2 days (1,911 cells/microliters) after surgery. By day 6, lymphocyte numbers were similar to preoperative values. Mean mononuclear cell count was 770 cells/microliters before surgery and was significantly increased on days 1 (3,084 cells/microliters), 2 (3,285 cells/microliters), and 6 (2,349 cells/microliters) after surgery. Mean eosinophil numbers were 1,388 cells/microliters before surgery and were significantly increased on day 6 (6,347 cells/microliters) only. Interaction between time and stage of gestation was found only for specific gravity and total protein concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Interactions between the monocyte/macrophage and the vascular smooth muscle cell. Stimulation of mitogenesis by a soluble factor and of prostanoid synthesis by cell-cell contact.

The effect of soluble factors from the monocyte/macrophage (M phi) on cell proliferation and the functional effects of cell-cell contact on the arachidonic acid (AA) cascade were studied with vascular smooth muscle cells (SMCs). Peripheral blood M phi s were isolated by adherence or in a Percoll gradient, and alveolar M phi s were obtained by lavage. Conditioned medium (CM) was prepared by preincubating M phi s with medium alone or by separating SMC and M phi cocultures by a membrane insert. Cell proliferation (image analysis) and 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha, radioimmunoassay) were measured in SMCs. Labeled prostanoids and other eicosanoid metabolites were isolated by high-performance liquid chromatography from SMCs prelabeled with 14C-AA. M phi s did not synthesize 6-keto-PGF1 alpha. The CM enhanced proliferation but did not stimulate 6-keto-PGF1 alpha synthesis in SMCs. However, cell-cell contact in cocultures of SMCs with the same concentration of M phi s used to generate CM resulted in increased 6-keto-PGF1 alpha synthesis by SMCs. Since the stimulatory effect of cell contact was not blocked by butylated hydroxytoluene, it could not be attributed to an oxidative burst from M phi s. Functional studies showed that the stimulatory effect of cell contact was enhanced by exogenous free AA and by endogenous AA release through A23187. Release of total radioactivity from prelabeled SMCs was enhanced by cell contact, and this effect was blocked by indomethacin (IM). Cell contact did not increase the release of free AA from prelabeled SMCs, even in the presence of IM. Finally, cell contact only stimulated the formation of prostanoids (IM-sensitive eicosanoid metabolites) from prelabeled SMCs. Lipoxygenase and other products of AA were not formed through cell-cell contact. These data showed that M phi s express a soluble factor that enhances SMC proliferation without affecting prostanoid synthesis. Subsequent cell contact between SMCs and M phi s stimulates prostanoid synthesis, which may possibly serve as a local and focal homeostatic mechanism for the regulation of uncontrolled SMC proliferation in atherogenesis.

Action of bradykinin at the cyclooxygenase step in prostanoid synthesis through the arachidonic acid cascade.

Bradykinin enhances prostanoid synthesis in aorta smooth muscle cells. Free arachidonic acid also enhances prostanoid synthesis and bradykinin, unlike fatty acid releasing agents, has a synergistic effect with free arachidonic acid. Bradykinin promotes metabolite release from cells prelabeled with [14C]-arachidonic acid and this effect is blocked completely by indomethacin. High performance liquid chromatography shows increase amounts of labeled 6-keto-prostaglandin F1 alpha, prostaglandin E2 and three additional cyclooxygenase-dependent metabolites but no increase in free arachidonic acid or other metabolites either in the absence or presence of indomethacin. Fatty acid releasing agents such as A23187 and cyclosporine A have very different effects on cells. These agents enhance levels of prostanoids, a number of other cyclooxygenase-independent metabolites, and free arachidonic acid which is even more elevated with added indomethacin. Bradykinin behaves in all respects like another agent, bacterial lipopolysaccharide, and the action of both agents is consistent with a mechanism involving cyclooxygenase rather than fatty release in the arachidonic acid cascade.

Effects of oxidized low density lipoproteins on arachidonic acid metabolism in smooth muscle cells.

The role of oxidized plasma lipoproteins in modifying arachidonic acid (AA) metabolism was studied in smooth muscle cells (SMC). Very low density lipoproteins (VLDL), unoxidized low density lipoproteins (LDLBHT) isolated with butylated hydroxytoluene (BHT), and oxidized LDL (LDLOXID) were separated from human serum. Thiobarbituric acid reactant (TBAR) levels were adjusted by saline incubations. Prostanoids in guinea pig SMC cultures were measured either by radioimmunoassay (RIA) or the isolation by high performance liquid chromatography (HPLC) of labeled prostanoids from SMC prelabeled with [14C]AA. Cell morphology and viability were studied by staining with Giemsa, nile red, and propidium iodide. VLDL and LDLBHT had little effect on prostanoid synthesis. Low-TBAR-LDLOXID enhanced total prostanoid levels and diminished the release of labeled prostanoids. Similar effects were found with exogenous free AA (unlabeled). Low-TBAR-LDLOXID did not affect the release of endogenous phospholipid AA as free AA. Synergism occurred between LDLOXID and exogenous free AA in prostanoid synthesis. Low-TBAR-LDLOXID evidently enhanced prostanoid levels in SMC both by supplying AA and by stimulating cyclooxygenase. High-TBAR-LDLOXID blocked prostanoid synthesis and enhanced cell death but time and pulse-recovery experiments showed that these effects were unrelated. High-TBAR-LDLOXID stimulated prostanoid synthesis when BHT was added to the incubation media. High-TBAR-LDLOXID also caused massive free AA release and the formation of many nonprostanoid derivatives. High-TBAR-LDLOXID evidently diminished overall prostanoid levels in SMC by inhibiting cyclooxygenase and at the same time stimulating AA release and the formation of other AA derivatives.

Bacterial lipopolysaccharide acts on human endothelial cells to enhance the adherence of peripheral blood monocytes.

The effects of bacterial lipopolysaccharide on the adherence of human peripheral blood monocytes (M0) to endothelial cells (ECs) were investigated in a quantitative adherence assay as an in vitro model of M0-EC interactions. ECs exposed for 2 hours or longer to 0.10 to 10 micrograms/ml lipopolysaccharide demonstrated significant increases in the levels of M0 adherence compared to control cells. By contrast, lipopolysaccharide added directly to adherence assays or preincubated with M0 did not increase M0 adherence to ECs. The effects of interleukin-1 (IL-1) were similar to the effects of lipopolysaccharide in that they acted solely on ECs to enhance M0 adherence and accelerated the rate of EC adherence to M0. Lipopolysaccharide did not increase EC adherence of M0 by causing damage or extracellular release of soluble mediators. The increase in M0-EC adherence by the chemotactic peptide formyl-methionyl-leucylphenylalanine (FMLP) was quite different from that by lipopolysaccharide and IL-1. FMLP rapidly induced M0 to become more adherent to ECs and plastic surfaces but was unable to act on ECs and enhance M0 binding. Thus M0-EC interactions are enhanced by the direct effects of FMLP on M0, whereas the actions of lipopolysaccharide and IL-1 are entirely focused on ECs and can be distinguished by differing characteristics and cellular targets.

Mechanisms for the stimulation of prostanoid synthesis by cyclosporine and bacterial lipopolysaccharide.

Both cyclosporine and bacterial lipopolysaccharide enhance prostanoid synthesis and regulate the immune response. This study was designed to establish whether these agents affect prostanoid synthesis by common or different mechanisms. CsA and LPS stimulate prostanoid synthesis both in human monocytes and smooth muscle cells from guinea pig aorta. Only LPS stimulates synthesis in the presence of exogenous arachidonic acid. Dexamethasone totally blocks CsA but only partially inhibits LPS. CsA and LPS both enhance the release of labeled metabolites from cells labeled with arachidonic acid, but indomethacin only blocks the effect of LPS. CsA and the releasing agent calcium ionophore (A23187) both increase PGE2 and PGI2 synthesis without changing their relative concentrations, cause the release of free arachidonic acid, and lead to the formation of new metabolites that are not products of cyclooxygenase activity. Preincubation with either CsA or A23187 and a subsequent wash deplete the arachidonic acid pool available for prostanoid synthesis. Thus, A23187 and CsA have very similar effects on arachidonic acid metabolism. In contrast, LPS increases PGE2 and PGI2 synthesis and alters their relative concentrations, diminishes the relative concentration of free arachidonic acid, and enhances the formation of new metabolites that are products of cyclooxygenase activity. These differences are explained by mechanisms in which CsA promotes prostanoid synthesis through arachidonic acid release, and LPS promotes prostanoid synthesis through increased cyclooxygenase activity.

Studies on oxidized low density lipoproteins. Controlled oxidation and a prostaglandin artifact.

Low density lipoproteins (LDL), isolated by ultracentrifugal flotation, were oxidized (LDLOXID) slowly during dialysis against 0.15 M NaCl and subsequent incubation in 96% air-4% CO2 at 37 degrees C. Butylated hydroxytoluene prevented LDL oxidation. LDL preparations from different sera were oxidized at different rates and the degree of lipid peroxidation was controlled by varying the incubation time. Mild oxidation did not alter the electrophoretic mobility of the LDLOXID preparations. LDLOXID contained lipid peroxides in neutral lipids, had increased amounts of lysophosphatidylcholine, and contained a number of complex oxidation products that were generated from the oxidation of free fatty acids. These oxidation products included large amounts of soluble material that cross-reacted with antibodies to PGE2 but not 6-keto-PGF1 alpha. The amount of cross-reacting material was proportional to the degree of lipid peroxidation. Cross-reacting material in LDLOXID preparations was evidently formed from the oxidation of free fatty acids released from LDL, since cross-reacting material was also formed when a synthetic fat emulsion was oxidized in the presence of free arachidonic acid.

Phase-front measurements of an injection-locked AlGaAs laser-diode array.

The phase-front quality of the primary spatial lobe emitted from an injection-locked gain-guided AlGaAs laser diode array is measured by using an equal-path, phase-shifting Mach-Zehnder interferometer. Root-mean-square phase errors of 0.037 +/- 0.003 wave (lambda/27) are measured for the single spatial lobe, which contained 240-mW cw output power in a single longitudinal mode. This phase-front quality corresponds to a Strehl ratio of S = 0.947, which results in a 0.23-dB power loss from the single lobe's ideal diffraction-limited power. These values are comparable with those measured for single-stripe index-guided AlGaAs lasers.

IL 1 and bacterial lipopolysaccharide increase the ability of human endothelial cells to bind peripheral blood monocytes.

Human endothelial cells (EC) exposed to human recombinant IL 1 and to bacterial lipopolysaccharide (LPS) demonstrated a time dependent increase in their ability to bind peripheral blood monocytes (M0). The enhancement of endothelial-M0 interactions was shown to represent a direct alteration of EC and was not explained by IL 1 and LPS affecting M0. Other experiments indicated that the enhancement represented an acceleration in EC binding of M0 suggesting that EC may play a role in initiating proinflammatory events prior to the recruitment of inflammatory cells by chemotactic peptides.

Effect of an oral dose of vitamin E on the vitamin E and cholesterol content of tissues of the vitamin E-deficient rat.

Weanling rats (female Sprague-Dawley) were fed until maturity a vitamin E-deficient diet or the deficient diet supplemented with 66 IU RRR-alpha-tocopheryl acetate/kg. Vitamin E, vitamin E quinone and total cholesterol levels in plasma, liver, paraovarian adipose tissue, lung, ovary and adrenal tissue were measured by high performance liquid chromatography. Vitamin E levels were greatly diminished, but cholesterol levels were unchanged in all tissues except adipose tissue of animals fed the deficient diet. Vitamin E-deficient animals received a single oral dose of 2 or 16.7 mg of RRR-alpha-tocopherol, and tissues were examined at 12 and 48 h. Plasma and liver formed a vitamin E pool that peaked at 12 h, had a high vitamin E/cholesterol ratio at 12 h and contained only trace amounts of vitamin E quinone. Adipose tissue, lung, ovary and adrenal concentrated vitamin E throughout the 48-h period, had low vitamin E/cholesterol ratios and contained small but significant amounts of vitamin E quinone. Vitamin E levels (micrograms/gram) at 48 h in lung, ovary and adrenal were higher than the vitamin E level in liver but the liver contained much more vitamin E (micrograms/organ) than the other tissues combined. Cholesterol levels (micrograms/gram) in plasma and liver decreased 45 to 55% in a dose- and time-dependent manner when a single oral dose of vitamin E was administered to deficient animals. Cholesterol levels in adipose tissue, lung and ovary were unchanged while the cholesterol level in adrenal increased 122% in a time-dependent manner with a single oral dose of vitamin E. These data show that a single oral dose of vitamin E has a profound effect on cholesterol levels in short-time experiments with the vitamin E-deficient rat. This rat model is appropriate for studies on the relationship between vitamin E and cholesterol metabolism in plasma, liver and the adrenal.

Deficiency of vitamin E in the alveolar fluid of cigarette smokers. Influence on alveolar macrophage cytotoxicity.

Cigarette smoking produces oxidant-mediated changes in the lung important to the pathogenesis of emphysema. Since vitamin E can neutralize reactive oxygen species and prevent peroxidation of unsaturated lipids, it may constitute an important component of the lung's defense against oxidant injury. To better characterize the antioxidant protective role of vitamin E, young asymptomatic smokers and nonsmokers were evaluated by bronchoalveolar lavage before and immediately after a 3-wk course of oral vitamin E (2,400 IU/d). Smoker alveolar fluid at baseline was relatively deficient in vitamin E compared with nonsmoker fluid (3.1 +/- 0.7 ng/ml vs. 20.7 +/- 2.4 ng/ml, P less than 0.005). Although smoker alveolar fluid vitamin E levels increased to 9.3 +/- 2.3 ng/ml after supplementation, the levels remained significantly lower than nonsmoker baseline levels (P less than 0.01). This deficiency was explained, in part, by the increased oxidative metabolism of vitamin E to the quinone form in the lungs of smokers compared with nonsmokers. Although the significance of a lower concentration of alveolar fluid vitamin E is unclear, it may compromise the antioxidant protection afforded by the alveolar fluid as it coats the lung's epithelial surface. The protective role of vitamin E was assessed by cytotoxicity experiments, which demonstrated that the killing of normal rat lung parenchymal cells by smoker alveolar macrophages was inversely related to the vitamin E content of the parenchymal cells. These findings suggest that vitamin E may be an important lower respiratory tract antioxidant, and that the deficiency seen in young smokers may predispose them to an enhanced oxidant attack on their lung parenchymal cells.