The effects of two forms of antioxidative co-therapy were analyzed in 24
interferon-alpha (IFN)-naive patients with chronic hepatitis C who were
randomized to either receive IFN monotherapy (3 x 4.5 million units IFN-alpha 2a per week), (group A), or IFN and N-acetylcysteine (N-acetylcysteine (NAC) 1.800 mg/day) plus sodium selenite (400 microg/day) supplementation (group B), or treatment as in group B plus vitamin E (544 IU/day) (group C), over 24 weeks. Changes in histology, normalization of ALT, reduction of viral RNA, serum levels of glutathione, selenium, vitamin E, erythrocyte glutathione peroxidase, trolox equivalent antioxidative capacity (TEAC), thiobarbituric acid reactive substances (TBARS) and protein carbonyl groups were measured. Low baseline TEAC and elevated TBARS indicated increased oxidative stress before therapy, which was not affected by antioxidant supplementation. At the end of treatment complete responses were found in 3/8, 2/8 and 6/8 patients in groups A, B and C, respectively, but liver histology had not significantly improved. Vitamin E treated patients had a 2.4 greater chance (95% CI: 1.05-5.5) of obtaining a complete response and had significantly greater reduction in viral load (P = 0.028) than patients without vitamin E. Relapses, i.e. re-appearance of
detectable hepatitis C virus (HCV) RNA and/or re-elevation of ALT-activity
occurred in 7 out of the 11 responders within 6 months after termination of
therapy (group A: 2/3, group B: 1/2 and group C: 4/6). Thus, no overall
beneficial effect of antioxidant/IFN therapy was detected. However, the apparent
trend towards a more favorable outcome with vitamin E supplementation warrants to further study this substance as an adjuvant to IFN therapy in chronic
hepatitis C.

To determine the independent contribution of specific immunologic and
nutritional factors on survival in HIV-1 disease, CD4 cell count, antiretroviral
treatment, plasma levels of vitamins A, E, B6, and B12 and minerals selenium and zinc were considered in relation to relative risk for HIV-related mortality.
Immune parameters and nutrients known to affect immune function were evaluated at 6-month intervals in 125 HIV-1-seropositive drug-using men and women in Miami, FL, over 3.5 years. A total of 21 of the HIV-1-infected participants died of HIV-related causes during the 3.5-year longitudinal study. Subclinical malnutrition (i.e., overly low levels of prealbumin, relative risk [RR] = 4.01, p < 0.007), deficiency of vitamin A (RR = 3.23, p < 0.03), vitamin B12
deficiency (RR = 8.33, p < 0.009), zinc deficiency (RR = 2.29.1, p < 0.04), and
selenium deficiency (RR = 19.9, p < 0.0001) over time, but not zidovudine
treatment, were shown to each be associated with HIV-1-related mortality
independent of CD4 cell counts <200/mm3 at baseline, and CD4 counts over time. When all factors that could affect survival, including CD4 counts <200/mm3 at baseline, CD4 levels over time, and nutrient deficiencies were considered jointly, only CD4 counts over time (RR = 0.69, p < 0.04) and selenium deficiency (RR = 10.8, p < 0.002) were significantly associated with mortality. These results indicate that selenium deficiency is an independent predictor of survival for those with HIV-1 infection.

Ageing is associated with impaired immune responses and increased
infection-related morbidity. This study assessed the effect of physiological
amounts of vitamins and trace elements on immunocompetence and occurrence of infection-related illness. 96 independently living, healthy elderly individuals
were randomly assigned to receive nutrient supplementation or placebo. Nutrient status and immunological variables were assessed at baseline and at 12 months, and the frequency of illness due to infection was ascertained. Subjects in the supplement group had higher numbers of certain T-cell subsets and natural killer cells, enhanced proliferation response to mitogen, increased interleukin-2 production, and higher antibody response and natural killer cell activity. These subjects were less likely than those in the placebo group to have illness due to infections (mean [SD] 23 [5] vs 48 [7] days per year, p = 0.002).
Supplementation with a modest physiological amount of micronutrients improves
immunity and decreases the risk of infection in old age.

To determine the impact of a trace element and vitamin supplementation on
infectious morbidity, a double-blind controlled trial was performed on 81
elderly subjects in a geriatric center during a 2-year period. Subjects were
randomly assigned to one of four treatment groups, and received daily: placebo;
trace elements/zinc 20 mg; selenium 100 micrograms); vitamins (vitamin C 120 mg; beta-carotene 6 mg; alpha-tocopherol 15 mg); or a combination of trace elements and vitamins at equal doses. (1) Before supplementation, low serum values in vitamin C, folate, zinc and selenium were observed in more than two thirds of the patients. (2) After 6 months of supplementation, a significant increase in vitamin and trace element serum levels was obtained in the corresponding treatment groups: a plateau was then observed for the whole study. (3) Subjects who received trace elements (zinc and selenium) alone or associated with vitamins had significantly less infectious events during the 2 years of supplementation. These results indicate that supplementation with low doses of vitamins and trace elements is able to rapidly correct corresponding
deficiencies in the institutionalized elderly. Moreover, zinc and selenium
reduced infectious events.

Excessive oxidative stress due to hyperglycemia and glycoxidation leads to an
increased production of F2-isoprostanes, one of which, 8-iso-PGF2 alpha, reaches
high concentrations in plasma and urine in both insulin-dependent and
non-insulin-dependent diabetics. This is associated with an increase in platelet
activation, reflected by an increased urinary excretion of platelet-derived
TxB2. Improved metabolic control or vitamin E supplementation reduces urinary
8-iso-PGF2 alpha and TxB2, whereas aspirin or indobufen reduces TxB2 but not
8-iso-PGF2 alpha. Since TxB2 in the urine seems to represent the common link
between diabetes (as well as other risk factors) and the thrombotic
complications of vascular disease, platelet activation due to
lipid-glycoxidation is an important aspect in the pathogenesis of vascular
complications of diabetes mellitus. Among the various plasma coagulation and
fibrinolysis factors that are found to be altered in diabetes, the increased
level of plasminogen activator inhibitor (PAI-1) in the plasma and in the vessel
wall is of the utmost importance. Indeed, it is suspected that the
atherosclerotic plaques formed in the presence of high concentrations of PAI-1
are more prone to rupture and ensuing thrombosis. The thrombosis-oriented
modifications of blood platelets, coagulation and fibrinolysis are an important
cause behind the high prevalence of vascular events in diabetes.

The present study evaluates the presence of oxidative stress in the uncontrolled
diabetic state. Glycemic control reduced the oxidative stress, but total
normalization of the parameters of oxidative stress was not achieved, indicating
continued oxidant injury despite optimal control of the diabetes. Vitamin E
supplementation for 4 weeks in these patients further reduced the oxidative
stress, suggesting that vitamin E supplementation might be helpful in reducing
free-radical-induced oxidant injury.

BACKGROUND: Diabetes mellitus is associated with increased generation of free
oxygen radicals and depleted scavenging potential (oxidative stress), leading to
increased LDL oxidation and platelet hyperreactivity, the major components of
atherothrombotic vascular lesions. A main goal of antioxidant therapy is to
protect the LDL particle from atherogenic oxidation and to reduce the activated
cellular hemostasis. METHODS: We evaluated the influence of a high dose
supplementation with 800 IU of the natural antioxidant RRR-alpha-tocopherol
(vitamin E) per day for six months on serum levels, vitamin E load of LDL
particles (HPLC), lag phase of LDL oxidation (Esterbauer's assay), platelet
adhesion molecules, leukocyte-platelet coaggregation (flow cytometry, D-III
protocol) and coagulation (INR/PTT) in a group of 36 patients with type 2
diabetes (f/m 22/14; age 588.0; HbA1 at baseline 10.251.7). RESULTS:
Average vitamin E levels increased 2.65-fold accompanied by a 1.83-fold increase
of LDL-associated vitamin E and a 12.3 min prolongation of the lag-phase of LDL
oxidation (p<0.001 for all parameters at six months). Platelet expression of
PECAM-1 (CD31) (-30.2% positive cells, p<0.001; antigen density -25%, p<0.001),
ICAM-2 (CD102) (-2.9% positive cells, p<0.01; antigen density -10.6%, p<0.001)
and fibrinogen (-1.6% positive cells, p<0.001; antigen density - 16.1 %,
p<0.001) decreased. Concomitantly, platelet-leukocyte-coaggregation increased by
44% (p<0.001), correlating to an INR reduction of 10.4% (1.060.09 to
0.950.09, p<0.001, r = - 0.34). The PTT remained constant. CONCLUSION: The
antioxidant protection from the increased vitamin E was accompanied by a
decreased expression of constitutive and function-dependent platelet adhesion
molecules. However, increases in platelet-leukocyte coaggregates and a shortened
INR time suggest extrinsic coagulation activation, possibly by induction of a
leukocyte tissue factor dependent mechanism. High dose supplements of
alpha-tocopherol may override the available redox balance in well controlled
type 2 diabetes. However, intrinsic effects of alpha-tocopherol must be
discussed.

Vascular disease accounts for the majority of the clinical complications in
diabetes mellitus. As an exaggerated oxidative stress degree has been postulated
as the link between diabetes mellitus and endothelial function, a possible
positive effect of plasma vitamin E (Vit.E) administration on brachial
reactivity could be postulated. Our study aims at investigating the possible
effect of chronic Vit.E administration on brachial reactivity, oxidative stress
indexes, and intracellular magnesium and calcium content in type II diabetic
patients free of diabetic complications. Forty adult, type II diabetic patients
were enrolled in the study, which was deigned as a double blind, randomized vs.
placebo trial. At baseline all patients underwent the following tests: 1)
anthropometric and metabolic examinations, 2) evaluation of oxidative stress
indexes, 3) intracellular magnesium and calcium measurements, and 4)
determination of arterial compliance and distensibility. Then, all patients were
randomly assigned to Vit.E treatment at a dose of 600 mg/day (Evion Forte; n =
20) or placebo (n = 20) over 8 weeks. At the end of this treatment period, a
complete reevaluation of the patients was made. Vit.E treatment was associated
with a significant improvement in the percent change in brachial artery diameter
(P<0.03) and oxidative stress indexes (P< 0.005). In the Vit.E group, the
percent change in brachial artery diameter correlated positively with the
percent change in oxidative stress indexes (oxidized/reduced glutathione,
Trolox-equivalent antioxidant capacity, thiobarbituric acid reaction products,
lipid peroxides) and intracellular cation content (magnesium and calcium). After
adjustment for age, sex, body mass index, and wait/hip ratio, all of these
correlations remained significant (P<0.03 for all). Furthermore, adjusting for
glycosylated hemoglobin, plasma total cholesterol, and homeostatic model index,
brachial artery diameter was still correlated with the percent change in
oxidative stress indexes (P<0.04 for all). Nevertheless, the relationship
between the percent change in brachial artery diameter and oxidative stress
indexes was no longer significant after adjustment for intracellular Mg and
Ca2+. In conclusion, our study demonstrates that chronic administration of Vit.E
improves brachial artery reactivity in patients with type II diabetes mellitus.
Such an effect seems mediated by a reduction in oxidative stress and a
regulation of intracellular calcium and magnesium contents.

Oxidative stress is produced under diabetic conditions and possibly causes
various forms of tissue damage in patients with diabetes. The aim of this study
was to examine the involvement of oxidative stress in the progression of
pancreatic beta-cell dysfunction in type 2 diabetes and to evaluate the
potential usefulness of antioxidants in the treatment of type 2 diabetes. We
used diabetic C57BL/KsJ-db/db mice, in whom antioxidant treatment
(N-acetyl-L-cysteine [NAC], vitamins C plus E, or both) was started at 6 weeks
of age; its effects were evaluated at 10 and 16 weeks of age. According to an
intraperitoneal glucose tolerance test, the treatment with NAC retained
glucose-stimulated insulin secretion and moderately decreased blood glucose
levels. Vitamins C and E were not effective when used alone but slightly
effective when used in combination with NAC. No effect on insulin secretion was
observed when the same set of antioxidants was given to nondiabetic control
mice. Histologic analyses of the pancreases revealed that the beta-cell mass was
significantly larger in the diabetic mice treated with the antioxidants than in
the untreated mice. As a possible cause, the antioxidant treatment suppressed
apoptosis in beta-cells without changing the rate of beta-cell proliferation,
supporting the hypothesis that in chronic hyperglycemia, apoptosis induced by
oxidative stress causes reduction of beta-cell mass. The antioxidant treatment
also preserved the amounts of insulin content and insulin mRNA, making the
extent of insulin degranulation less evident. Furthermore, expression of
pancreatic and duodenal homeobox factor-1 (PDX-1), a beta-cell-specific
transcription factor, was more clearly visible in the nuclei of islet cells
after the antioxidant treatment. In conclusion, our observations indicate that
antioxidant treatment can exert beneficial effects in diabetes, with
preservation of in vivo beta-cell function. This finding suggests a potential
usefulness of antioxidants for treating diabetes and provides further support
for the implication of oxidative stress in beta-cell dysfunction in diabetes.

OBJECTIVE: This study compared susceptibility to oxidation of low-density
lipoproteins (LDL) of non-diabetic and diabetic (Type 2) men and examined the
response of diabetic men to antioxidant supplementation (alpha-tocopherol,
beta-carotene and ascorbate). RESEARCH DESIGN AND METHODS: Twenty adult
non-diabetic and 20 diabetic men were recruited. Oxidation of LDL was assessed
by four different assay systems, and the extent of oxidation was assessed by
four different measurements. Diabetic men received eight weeks of placebo
("baseline"), twelve weeks of antioxidant supplements ("treated") and eight
weeks of placebo ("post-treatment"). Supplements provided 24 mg of
beta-carotene, 1000 mg of ascorbate and 800 IU of alpha-tocopherol daily.
RESULTS: With Cu oxidation at 37 degrees C, thiobarbituric reactive substances
(TBARS) formation was significantly higher (p=0.032) and loss of free amine
groups was significantly greater (p=0.013) in the LDL from diabetic subjects
than controls. Antioxidant supplementation of diabetic subjects significantly
decreased all parameters of LDL oxidation with Cu at 30 degrees C and 37 degrees
C. At 30 degrees C the lag phase increased from 55 to 129 minutes (p<0.0001);
conjugated diene formation decreased from 1.23 to 0.62 OD units (p<0.0001);
TBARS formation decreased from 78 to 33 nmoles MDA/mg LDL protein (p<0.0001);
and free amine loss decreased from 41 to 12% (p<0.0001). With Cu oxidation at 37
degrees C, similar changes occurred. CONCLUSIONS: These studies indicate that
the LDL from diabetic subjects are more susceptible to oxidation than LDL from
non-diabetic subjects. Supplementation of diabetic subjects with antioxidant
vitamins significantly decreases susceptibility of LDL to oxidation by Cu. These
studies are consistent with epidemiological and intervention studies suggesting
that antioxidant vitamin use significantly decreases risk for coronary heart
disease.