Peripheral biomarkers of oxidative stress and their limited potential in evaluation of clinical features of Huntington's patients.

CONTEXT: Peripheral oxidative biomarkers could be useful for monitoring clinical features of Huntington's disease (HD). MATERIALS AND METHODS: Cu/Zn-superoxide dismutase (Cu/Zn-SOD), neuron-specific enolase (NSE) and 8-hydroxy-2'-deoxyguanosine (8-oxoGua) serum levels were analysed in 18 HD patients and 10 controls. Clinical measures were recorded from each HD patients. RESULTS: Cu/Zn-SOD, NSE and 8-oxoGua values were higher in HD patients than in controls. Cu/Zn-SOD and NSE correlated positively. No correlation was observed between the biomarkers analysed and the clinical measures assessed. DISCUSSION AND CONCLUSION: Serum oxidative biomarkers could express the neuronal oxidative processes going on in HD patients but are inadequate to evaluate clinical features of the disease.

Inflammation and endothelial activation in benign prostatic hyperplasia and prostate cancer.

PURPOSE: Emerging insights underline a link among chronic inflammation and endothelial activation with benign prostatic hyperplasia (BPH) and prostate cancer (PCa). We aim to investigate whether specific plasma markers of inflammation and endothelial activation allow to discriminate BPH and PCa. MATERIALS AND METHODS: Fifteen patients affected by BPH, 15 by PCa and 15 controls, were enrolled. Interleukin-6 (IL-6), CD40 ligand (CD40L), endothelial-selectin (E-selectin), platelet-selectin (P-selectin), vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) were measured. RESULTS: In systemic blood samples, IL-6 has been found increased in patients affected by BPH (4.25 ± 0. pg/mL) and PCa (5.08 ± 0.24) respect to controls (2.62 ± 0.34; p < 0.05). CD40L was higher in BPH (4.25 ± 0.65 ng/mL; p < 0.05) than in control (2.31 ± 0.20) and PCa group (2.60 ± 0.56). E-selectin, P-selectin and VCAM-1 did not show any significant difference. Higher levels of ICAM-1 were detected in patients with PCa (573.04 ± 52.23) and BPH (564.40 ± 74.67) than in the controls (215.30 ± 11.53 ng/mL; p < 0.05). In local blood samples, IL-6 has been found significantly increased in PCa in comparison with patients with BPH; there was no difference in CD40L, E-selectin, P-selectin, VCAM-1 ed ICAM-1. CONCLUSIONS: Changes in inflammation and endothelial activation markers may be not considered to be of value in discriminating BPH and PCa.

Oxidative stress in benign prostatic hyperplasia and prostate cancer.

OBJECTIVE: The aim was to verify whether oxidative stress could represent a common key factor of benign prostatic hyperplasia (BPH) and prostate cancer (PCa). SUBJECTS AND METHODS: 15 patients affected by BPH, 15 with PCa and 15 controls were enrolled. Blood samples were withdrawn systemically and locally during radical retropubic prostatectomy in patients with PCa and during transvesical retropubic adenomectomy in patients diagnosed with BPH. Plasma oxidized low-density lipoprotein, peroxides, and total equivalent antioxidant capacity (TEAC) including plasma superoxide dismutase (SOD) determination were analyzed as oxidative markers. RESULTS: With respect to the control group, high plasma peroxides and decreased TEAC levels were measured in patients affected by both PCa and BPH. Plasma peroxides were significantly higher in patients with PCa with respect to BPH. A positive correlation was found between peroxides and TEAC values in samples withdrawn locally in patients affected by PCa. An inverse correlation between peroxides and TEAC was observed in patients with BPH. No statistically significant modifications were observed as concerns SOD activity and LDL oxidability. CONCLUSIONS: Our findings confirm a significant unbalance of redox status in patients affected by BPH and PCa, and suggest a potential involvement of oxidative stress as a determinant in the pathogenesis of these diseases.

Detrimental effects of anabolic steroids on human endothelial cells.

The aim of this study is to investigate the effects in vitro induced by androgenic anabolic steroids (AAS) (testosterone, nandrolone, androstenedione, norandrostenedione, and norandrostenediol) used illicitly in sport competitions, on the proliferation ability, apoptosis and the intracellular calcium concentration ([Ca2+]i) in human umbilical vein endothelial cells (HUVECs), selected as a prototype of a biological target system whose structure and function can be affected by steroids. For this purpose, we evaluated the proliferation inhibition by cytotoxic assay expressed as the concentration of drug inducing a 50% decrease in growth (IC50). The IC50 was reached for testosterone at 100 microM, androstenedione at 375 microM, nandrolone at 9 microM, norandrostenedione at 500 microM. The IC50 value for norandrostenediol was not reached until a concentration of 6000 microM. The apoptotic effect was evaluated by flow cytometry at IC50 for each drug. We observed that testosterone induced 31% of apoptotic cells, norandrostenedione 25%, androstenedione 15% and nandrolone 18%. We have analyzed the effects of these drugs on [Ca2+]i both in the immediate and long-term continuous presence of each compound. Our data show a statistically significant increase of [Ca2+]i in the acute condition and in long-term treated cultures, suggesting that androgen steroids modulate intracellular levels of calcium independent of incubation time or compound identity. As a whole, this study demonstrates that AAS might alter endothelial homeostasis, predisposing to the early endothelial cell activation that is responsible for vascular complications observed frequently in AAS users.

Impaired plasma nitric oxide availability and extracellular superoxide dismutase activity in healthy humans with advancing age.

This study is aimed to verify the modifications of extracellular superoxide dismutase (EC-SOD) activity and its potential involvement on the mechanism responsible for the impairment of plasma nitric oxide (NO) availability occurring with advancing age in healthy humans. For this purpose, plasma samples were drawn from 40 healthy men, aged 20-92 years, in fasting state and used for measurements of stable end-product nitrite/nitrate (NOx), as expression of NO availability, EC-SOD activity, thiobarbituric acid reactive substances (TBARS) as marker of lipid peroxidation, Trolox equivalent antioxidant capacity (TEAC) as a measure of plasma total antioxidant capacity, and in vitro susceptibility of low density lipoprotein (LDL) to copper-mediated oxidation, evaluated as lag time. As indicated by our results, advancing age was significantly related to decreased plasma values of NOx (r = -0.877, P < 0.001), EC-SOD activity (r = -0.888, P < 0.001), TEAC (r = -0.647, P < 0.001) and lag time (r = -0.621, P < 0.001) as well as to an increased plasma amount of TBARS (r = 0.858, P < 0.001). NOx plasma level resulted independently predicted by EC-SOD activity and age. EC-SOD activity, in turn, was determined by age and TEAC. Taken together, findings of the present study give further insight into the mechanism related to age-associated endothelial dysfunction, indicating that the decreased EC-SOD activity may be involved in the progressive reduction of plasma NO availability with advancing age through the age-related impairment of oxidant/antioxidant balance.

Mean Platelet Volume During Ischemic Stroke is a Potential Pro-inflammatory Biomarker in the Acute Phase and During Neurorehabilitation Not Directly Linked to Clinical Outcome.

The prognostic role of increased mean platelet volume (MPV), as an indicator of platelet activation and large, more reactive platelets, in clinical and functional outcome of ischemic stroke is still conflicting. Studies are not currently available on the association between MPV and stroke recovery after neurorehabilitation. The relationship between MPV and clinical and functional outcome measures was assessed in twenty-four patients in the acute phase of first-ever ischemic stroke, and before and after 8-week intensive multifunctional neurorehabilitation. Neurorehabilitation was associated with improved scores of the National Institutes of Health Stroke Scale (NIHSS), the modified Rankin Scale (mRS), and the modified PULSES profile (mPULSES). When compared with apparently healthy subjects, higher MPV values were observed in stroke patients 24 hours after stroke and before neurorehabilitative treatment started not later than 14 days after stroke. Decreased MPV values were found after neurorehabilitation, even if the absolute values were still higher than those detected in control subjects. No correlation was observed between MPV values and scores of the NIHSS and mRS scales evaluated in stroke acute phase. No correlation was also observed before and after neurorehabilitative treatment between MPV and NIHSS, mRS and mPULSES scores. Our data provide evidence of the effectiveness of neurorehabilitation on modulating MPV values and support the hypothesis that high MPV could represent an expression of proinflammatory condition of the stroke patients, realistically pre-existent to acute ischemic event, than a marker of neurologic deficit and disability or of stroke recovery including motor performance and functional independence.

Uric acid and Cu/Zn superoxide dismutase: potential strategies and biomarkers in functional recovery of post-acute ischemic stroke patients after intensive neurorehabilitation.

No evidence is currently provided on the involvement of uric acid (UA) and Cu/Zn Superoxide Dismutase (Cu/Zn SOD) in functional recovery of stroke patients after neurorehabilitation. For this purpose, the relationship between UA and Cu/Zn SOD plasma levels and clinical and functional outcome measures were analysed in twenty-five post-acute stroke patients undergoing intensive neurorehabilitation. UA and Cu/Zn SOD plasma levels were evaluated in fifteen healthy subjects as control values. Neurorehabilitation was associated with improved scores (P<0.05) of the National Institutes of Health Stroke Scale (NIHSS), the modified Rankin Scale (mRS), and the mPULSES profile. UA plasma levels were higher before neurorehabilitation, decreased after, but were still higher than control values. Conversely, Cu/Zn SOD plasma levels were lower than control values before neurorehabilitation and increased after, even though the absolute values were still lower than controls. An inverse correlation was found between variations of UA plasma levels observed before and after neurorehabilitation (Δ UA) and those of Cu/Zn SOD (Δ Cu/Zn SOD) (r= -0.386; P<0.001). No significant correlations were observed between ΔUA and the variations of the scores observed in all clinical outcome measures before and after neurorehabilitation (Δ scores of clinical outcome measures). Δ Cu/Zn SOD correlated positively with Δ NIHSS, Δ mRS and Δ mPULSES scores. Our data provide evidence of neurorehabilitation effectiveness on modulating UA and Cu/Zn SOD plasma levels and suggest that Cu/Zn SOD could assume the significance of biomarker of functional recovery, rather than UA that could be a marker of the magnitude of oxidative injury.

Influence of intensive multifunctional neurorehabilitation on neuronal oxidative damage in patients with Huntington's disease.

The influence of intensive multifunctional neurorehabilitation on serum levels of Cu/Zn-superoxide dismutase (Cu/Zn-SOD), neuron-specific enolase (NSE), and 8-hydroxy-2-deoxyguanosine (8-OHdG), as markers of oxidative damage, was evaluated in symptomatic patients with Huntington's disease (HD). Improved clinical outcome measures were observed after neurorehabilitation. Baseline levels of Cu/Zn-SOD, NSE and 8-OHdG were higher than those observed in controls. Cu/Zn-SOD and NSE values decreased after neurorehabilitation, but were still higher than those measured in controls. Cu/Zn-SOD and NSE correlated positively before (r=0.659; p=0.003) and after rehabilitation (r=0.553, p=0.017). 8-OHdG values decreased after neurorehabilitation without reaching significance when compared with baseline values (p=0.145). No correlation was observed between the measured oxidative markers and the assessed clinical outcome measures, either before or after neurorehabilitation. The findings reported in the present paper provide evidence of the effectiveness of neurorehabilitation in reducing oxidative damage in HD patients and underline the limit of serum oxidative markers for the evaluation of clinical features of HD.

Oxidative stress in post-acute ischemic stroke patients after intensive neurorehabilitation.

We investigated in post-acute ischemic stroke patients the influence of intensive neurorehabilitation on oxidative stress balance during recovery of neurological deficits. For this purpose, fourteen patients were included in the study within 30 days of stroke onset. Outcome measures were the National Institutes of Health Stroke Scale (NIHSS), the modified Rankin Scale (mRS), the Barthel Index, and the Katz Index. Redox balance was assessed by measuring plasma peroxidative by-products, nitrite/nitrate metabolites (NOx), as an index of nitric oxide (NO), Cu/Zn Superoxide Dismutase (Cu/Zn SOD) activity, serum urate concentration, autoantibodies against ox-LDL (OLAB) serum level and plasma antioxidant capacity. Assessments were made before and after neurorehabilitation. Fifteen apparently healthy controls were investigated to compare redox markers. Intensive neurorehabilitation was associated with an improvement of all the outcome measures (P < 0.05). Decreased values of peroxidative by-products and of NOx (P < 0.05) were observed after neurorehabilitation in stroke patients even though their values were higher than in controls (P < 0.05). Changes observed before and after neurorehabilitation in NIHSS scores (Δ NIHSS scores) and in plasma NOx amount (Δ NOx) correlated positively (r=0.79; P < 0.005). No differences in EC-SOD activity, OLAB and serum urate concentrations were found between stroke patients and controls, before and after neurorehabilitation. Total plasma antioxidant capacity, lower in stroke patients than in controls before neurorehabilitation, was unchanged thereafter. Our data provide evidence of the effectiveness of neurorehabilitation on reducing redox unbalance in stroke patients and hints the role of NO as a messenger involved in post-ischemic neuronal plasticity influencing recovery of neurological deficits.

Evidence of redox unbalance in post-acute ischemic stroke patients.

Measurements of the redox balance after the ischemic stroke occurrence might be useful to monitor the outcome of patients who suffered an ischemic stroke in terms of stroke recurrence and other vascular events. For this purpose, fifteen patients (mean age 71.40±2.50 years) with a first-ever ischemic stroke were included in the study within 30 days of stroke onset. Stroke severity was evaluated according to the National Institutes of Health Stroke Scale (NIHSS). Redox balance was assessed by measuring plasma amount of total peroxidative by-products, nitrite/nitrate metabolites (NOx), as expression of nitric oxide (NO) plasma bioavailability, total plasma antioxidant capacity (TEAC), Cu/Zn Superoxide Dismutase (Cu/Zn SOD) activity, serum urate concentration and autoantibodies against ox-LDL (OLAB) serum level. Total cholesterol, triglycerides, high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were also measured. Fifteen apparently healthy controls (mean age 70.28±2.03 years) were investigated to compare redox markers. Stroke patients had higher plasma values of total peroxidative by-products, NOx stable metabolites and of total cholesterol, triglycerides, and LDL-C than controls (P < 0.05). No differences in OLAB levels, Cu/Zn-SOD activity, serum urate concentration, and plasma HDL-C amount were found in stroke patients when compared to controls. Total plasma antioxidant capacity was lower in stroke patients than in controls. NOx values correlated positively with the NIHSS score in stroke patients (r=0.668; P=0.0065). The observed presence of redox unbalance in stroke patients could represent an early indicator of diffuse endothelial activation during which patients may be at increased risk of stroke recurrence and other vascular events.

Moderate consumption of red wine and human platelet responsiveness.

Available studies showed an inverse association between red wine consumption and prevalence of vascular risk factors in coronary hearth disease and stroke. Effects were mainly associated to wine antioxidant and antiaggregant properties. Actually, in vitro studies indicate a favourable effect of wine and/or of its non-alcoholic components in decreasing platelet sensitivity and aggregability. In a 4-week supplementation in 15 healthy male volunteers, we evaluated whether moderate red wine consumption might improve antioxidant defence mechanisms and promote positive modulation of inflammatory cytokines and cell adhesion molecules in relation to platelet responsiveness. We did not find any change of ADP- and collagen-induced platelet aggregation ex vivo, any change of biomarkers of oxidative stress, and any change of plasma lipid profile and haemostatic parameters, with the only exception of decreased fibrinogen levels (P<0.05). We also found an increase of mean platelet volume (P<0.05) without any significant modification of CD40 Ligand and P-selectin levels. Increased expressions of intercellular adhesion molecule-1, soluble E-selectin and interleukin-6 (P<0.05) were also observed. According to our findings increased circulating levels of inflammatory and endothelial cell activation markers may indicate a low-grade systemic inflammation and vascular activation that could be responsible for the lack of inhibition or of decreased platelet responsiveness, possibly because the plasmatic increase of wine antioxidant compounds is insufficient to improve endothelial function and to counteract the influence of ethanol on endothelial activation.

Preventive non-pharmacological treatment and nitric oxide in chronic migraine.

In chronic migraine the central sensitisation and the changes of regional cerebral blood flow are mediated by nitric oxide (NO) and oxygen free radicals. Biofeedback is considered a preventive non-pharmacological treatment decreasing migraine attacks. We investigated whether biofeedback effectiveness is related to relaxation processes and its influence on oxidative stress. The Migraine Disability Assessment Score (MIDAS) and serum NO stable metabolites (NO(x)) were evaluated in 20 patients with chronic migraine before and at the end of biofeedback sessions. MIDAS score was lower after biofeedback than that reported before treatment. NO(x) serum levels were higher after biofeedback than those measured before starting treatment. Thus, the effectiveness of biofeedback is related to a muscular relaxation and to its influence on NO bioavailability in patients with chronic migraine.