Glutathione, vitamin E and oxidative stress in coronary artery disease: relevance of age and gender

Background  Observational studies suggest that low levels of antioxidants are associated with high risk for coronary artery disease (CAD). We investigated whether the biomarkers of oxidative balance undergo the same modifications in all CAD patient groups, regardless of gender and age.
Materials and methods  One hundred sixty-eight CAD patients and 107 healthy controls were assayed for plasma levels of reduced glutathione (GSH), α- and γ-tocopherol (α- and γ-T) as endogenous antioxidants. A damage score (DS), representative of oxidative stress status, was calculated. ancova models were used to test the association between antioxidants, DS and CAD and its modulation by age and gender.
Results  The DS was higher in CAD than in controls. GSH levels, were lower in CAD patients (mean ± SEM: 57·61 ± 1·87 μmol 10 g⁻¹ haemoglobin vs. 68·55 ± 2·23 in controls, P  < 0·0006) in males and in older subjects. Levels of other antioxidants exhibited a complex pattern. Overall, no difference was found in α- and γ-T contents between CAD and controls, but lower α-T values were observed in CAD females. A significant interaction between CAD status and gender was observed ( P  = 0·003).
Conclusions  Our study shows that the involvement of antioxidants in CAD is related to patients' characteristics. These findings may be relevant in planning antioxidant therapies.     

Similar increase in oxidative stress after fat overload in persons with baseline hypertriglyceridemia with or without the metabolic syndrome

Objective

We compared the levels of biomarkers of oxidative stress before and after a fat overload in three groups.

Materials and methods

17 controls and two groups with hypertriglyceridemia: 43 without the metabolic syndrome (TG-non-MS) and 29 with the metabolic syndrome (TG-MS). All subjects underwent a 60 g fat overload. Baseline measurements included glucose, BMI (body mass index), waist circumference and HOMA IR (homeostasis model assessment insulin resistance). Cholesterol, triglycerides, HDL (high density lipoprotein) cholesterol, TNF-alpha (tumor necrosis factor) and IL-6 (interleukin-6), lipoperoxide (LPO), carbonylated proteins, reduced glutathione (GSH), oxidized glutathione (GSSG), glutathione peroxidase (GSH-PX), glutathione reductase (GSH-Rd), catalase and glutathione transferase (GST) were measured at baseline and 3 h after fat overload.

Results

Compared to the controls, the two patient groups had higher plasma levels at baseline and after overload of cholesterol, triglycerides and apolipoprotein B, LPO, carbonylated proteins and GSSG, and lower levels of antioxidants at baseline and after the fat overload.

Conclusion

The two patient groups had the same degree of oxidative stress.     

Inter-individual variability of plasma PAF-acetylhydrolase activity in ARDS patients and PAFAH genotype

Background:  Platelet activating factor (PAF), a pro-inflammatory phospholipid, stimulates cytokine secretion from polymorphonuclear leukocytes expressing the transmembrane G-protein coupled PAF receptor. Elevated PAF levels are associated with acute respiratory distress syndrome (ARDS) and sepsis severity. The pro-inflammatory effects of PAF are terminated by PAF acetylhydrolase (PAF-AH).
Objective:  We sought to determine whether allelic variants in the human PAFAH gene (Arg92His, Ile198Thr, and Ala379Val) contribute to variability in PAF-AH activity in patient plasma obtained within 72 h of ARDS diagnosis.
Results:  Plasma PAF-AH activity (mean ± SD) was higher in patients homozygous for the Arg92 allele compared to His92 allele carriers (2·21 ± 0·77 vs. 1·64 ± 0·68 U/min; P  < 0·01; n  = 31 and 21 respectively). Baseline plasma PAF-AH activity was higher among day 7 survivors vs. day 7 non-survivors (2·05 ± 0·75 vs. 1·27 ± 0·63, P  = 0·05).
Conclusion:  These data demonstrate an association between PAF-AH allelic variation, plasma activity, and outcome in ARDS.     

Evaluation of oxidative stress and inflammation in obese adults with metabolic syndrome.

BACKGROUND: Obesity and metabolic syndrome increase the risk of cardiovascular morbidity and mortality. Oxidative stress seems to be involved in the pathophysiology of diabetes and cardiovascular complications of metabolic syndrome. The aim of our study was to evaluate the level of oxidative stress and inflammation in obese adults with and without metabolic syndrome. METHODS: Oxidative stress and inflammation markers (total amount of free radicals, malondialdehyde, allantoin, alpha1-antiproteinase, oxidized/reduced glutathione ratio, high-sensitive C-reactive protein, fibrinogen), total antioxidant capacity and lipid standardized alpha-tocopherol were determined in obese subjects fulfilling at least three criteria of metabolic syndrome according to the National Cholesterol Education Program-Adult Treatment Panel III guidelines (n=20 patients), in obese subjects without metabolic syndrome (n=20 patients) and in 48 healthy controls. RESULTS: Oxidative stress and inflammation markers were significantly elevated in the obese subjects, especially in those exhibiting metabolic syndrome. According to multidimensional statistical analysis, oxidative stress was independently related to triacylglyceride concentration, abdominal fat, low high-density lipoprotein cholesterol and low lipid standardized alpha-tocopherol in the patients with metabolic syndrome. CONCLUSIONS: High levels of free radicals together with low antioxidant capacity detected in obese adults indicate elevated oxidative stress, which is--together with systemic inflammation--further potentiated in the case of obese patients with metabolic syndrome. This imbalance in oxidative/antioxidative status and subclinical inflammatory state leads to higher risk of atherosclerotic and diabetic complications.     

Estimated liver weight is directly related to hepatic very low-density lipoprotein-triglyceride secretion rate in men

Background  Animal studies suggest that liver weight is directly related to hepatic very low-density lipoprotein–triglyceride (VLDL-TG) secretion, independently of body size. This relationship has never been examined in humans.
Materials and methods  We measured VLDL-TG secretion rate by using stable isotope-labelled tracers in 21 healthy, non-obese men (age: 25 ± 3 years; body mass index: 24·8 ± 1·6 kg m⁻²), and evaluated the relationship between VLDL-TG secretion and indices of total and regional adiposity (body mass index, total body fat, trunk fat), metabolic parameters (free fatty acid, glucose, and insulin concentrations, homeostasis model assessment index of insulin resistance, resting energy expenditure), and estimated liver weight.
Results  Correlation analysis showed that estimated liver weight was positively associated with total VLDL-TG secretion rate ( r  = 0·722, P  < 0·001), VLDL-TG secretion rate per liter of plasma ( r  = 0·562, P  = 0·008), VLDL-TG secretion rate per kilogram of body weight ( r  = 0·555, P  = 0·009), and VLDL-TG secretion rate per kilogram of liver weight ( r  = 0·620, P  = 0·003). In multiple regression analysis, estimated liver weight was the only significant predictor of VLDL-TG secretion rate regardless of units of expression, explaining 31–52% of total variance; none of the metabolic parameters and indices of body fatness entered the regression models.
Conclusions  We conclude that estimated liver weight is directly related to hepatic VLDL-TG secretion rate in healthy non-obese men; this relationship is likely not mediated by interindividual variation in body size.     

Clinical relevance of measuring colonic permeability

Background  Gastroduodenal and small intestinal permeability are increased in patients with Crohn's disease (CD) and intensive care patients. The relevance of colonic permeability has not yet been adequately investigated. The aim of this study was to investigate the clinical value of sucralose excretion as indicator for colonic permeability in these patient groups.
Design  After oral administration of four sugars and subsequent analysis of urinary excretion, gastroduodenal and intestinal permeability were calculated from saccharose excretion and lactulose/mannitol (L/M) ratio over 5 h, and sucralose excretion from 5 to 26 h in 100 healthy controls, 29 CD and 35 patients after coronary surgery (CABG).
Results  In controls, sucralose excretion was highly variable (0·67 ± 0·92%) and not related to small intestinal permeability. In CD and CABG, L/M ratio was increased (0·054 ± 0·060; 0·323 ± 0·253 vs. 0·018 ± 0·001 in controls). Sucralose excretion was increased in 77% of CABG but only in 7% of CD. There was an association between gastroduodenal and intestinal permeability in CD and CABG ( r  = 0·72, and r  = 0·51), but sucralose excretion was not related to either one of these two parameters. Other than a weak association between sucralose and length of stay in intensive care in CABG patients ( P  = 0·099), sucralose excretion was not related to clinical outcome.
Conclusions  The proposed cut-off for normal sucralose excretion is 2·11%, but its high variability and lack of association to gastrointestinal permeability or clinical outcome leave it open, if it can provide information beyond established permeability tests.     

High extracellular induced sputum haem oxygenase-1 in sarcoidosis and chronic beryllium disease

Background  Sarcoidosis and chronic beryllium disease (CBD) are inflammatory conditions in which oxidative stress state may be crucial for disease outcome. This study compares haem oxygenase-1 (HO-1) extracellular activity for the first time in patients with sarcoidosis or CBD and in healthy controls.
Materials and methods  Induced sputum was recovered using a standard protocol. Pulmonary function tests (PFT) were performed by conventional methods. T lymphocyte subsets (CD4 and CD8) were measured by flow activated cell shorter (FACS). The HO-1 and nitrite levels were measured by a bilirubin-biliverdin reductase-dependant reaction and Greiss reaction respectively. Ferritin and iron levels were measured by enzymatic reaction and chemiluminometric immunoassay respectively.
Results  The mean percentage of lymphocytes was significantly higher in the 36 sarcoid patients compared with that in the 17 controls ( P  =   0·001). The mean CD4/CD8 ratio was significantly higher in the sarcoid and the 10 CBD patients compared to that in controls ( P  =   0·000 and 0·002 respectively), as was the mean HO-1 activity ( P  =   0·045 and 0·041 respectively). The HO-1 activity did not differ with the sarcoidosis stage. The HO-1 level and PFT parameters were negatively correlated. The differences in mean nitrite, ferritin and iron levels were non-significant between the three groups. The HO-1 and ferritin levels were correlated ( P  =   0·008).
Conclusions  We succeeded in non-invasively measuring the activity of HO-1 from cells of airways in spite of its being an intracellular enzyme. The HO-1 levels in sarcoidosis and CBD were abnormally elevated.     

Selective effects of CPAP on sleep apnoea-associated manifestations

Background   Visceral adiposity and obstructive sleep apnoea (OSA) may be independently associated with daytime sleepiness/low performance, insulin resistance, hypercytokinaemia, and/or hypertension. The objectives of this study are to simultaneously test these associations at baseline and after 3 months of continuous positive airway pressure (CPAP) therapy.
Materials and methods   Sixteen obese men with OSA; 13 non-apnoeic, obese controls, and 15 non-obese controls were monitored in the sleep laboratory for four consecutive nights. Objective measures of daytime sleepiness and performance, serial 24 h plasma measures of interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), TNF receptor 1 (TNF-r1) and adiponectin, fasting blood glucose and insulin, visceral adiposity and blood pressure were obtained. Sleep apnoeics were re-assessed using the same protocol after 3 months of CPAP.
Results   At baseline, IL-6, TNF-r1, and insulin resistance were highest in OSA patients, intermediate in obese controls, and lowest in non-obese controls ( P  < 0·05). Visceral fat was significantly greater in sleep apnoeics than obese controls and predicted insulin resistance and IL-6 levels, whereas OSA predicted TNF-r1 levels ( P  < 0·05). CPAP decreased daytime sleepiness and blood pressure ( P  < 0·05), but did not affect fasting glucose or insulin or around the clock adiponectin, IL-6, TNF-α, or TNF-r1 levels.
Conclusions   In obese sleep apnoeics, visceral fat is strongly associated with insulin resistance and inflammation. CPAP decreases sleepiness and moderates hypertension but does not affect visceral adiposity, insulin resistance, hypoadiponectinaemia or hypercytokinaemia, all of which are independent risk factors for cardiovascular disease and diabetes.     

The hormone sensitive lipase gene in familial combined hyperlipidemia and insulin resistance

Backround Insulin resistance in the most common familial dyslipidemia, familial combined hyperlipidemia (FCHL), could be due to variations in the hormone sensitive lipase (HSL) gene.
Materials and methods The coding region of the HSL gene was screened with the single strand conformation polymorphism analysis in probands of 27 FCHL families with 228 members. In addition, the C-60G promoter substitution of the HSL gene was determined by the restriction fragment length polymorphism analysis in these subjects.
Results No variants in the coding region of the HSL gene were found and the allele frequencies of the C-60G promoter substitution and the silent variant (G3138A) in the 3' untranslated region did not differ between 110 control subjects and 27 probands with FCHL. However, in control women the C-60G substitution was associated with high body mass index [30·6 ± 0·9 kg m⁻² (mean ± SD) in subjects with the C/G genotype and 24·8 ± 4·6 in subjects with the C/C genotype, P  = 0·012], and in control men with high rates of insulin-stimulated whole body glucose uptake (70·1 ± 14·7 vs. 56·7 ± 14·2 µmol kg⁻¹ min⁻¹, P  = 0·014). In 228 FCHL family members this substitution was associated with high low-density lipoprotein cholesterol levels in men (4·51 ± 1·12 vs. 5·17 ± 1·28 mmol L⁻¹, P  = 0·049), but not in women.
Conclusions The HSL gene is not a major gene for FCHL. However, the − 60G allele of this gene may affect body weight, insulin sensitivity and serum cholesterol levels.     

Low leptin levels in migraine: a case control study

Background.— Obesity has been shown to be a risk factor for transformation of episodic migraine to chronic form, and adipocytokines have been implicated to modulate some of the cytokins such as interleukin-6 and tumor necrosis factor, which also act in the neurogenic inflammation in migraine. The aim of the study was to assess leptin levels, one of the adipocytokines, in headache-free period of migraine patients and investigate its relation to vascular risk factors.
Material and Methods.— Sixty-one patients with episodic migraine headaches and 64 control subjects were enrolled in the study. Demographic data and anthropometric measurements were obtained from all participants; body mass index and fat mass values were calculated. Glucose and lipid parameters were measured by oxidase technique and cholesterol esterase enzymatic assays, and leptin levels were measured by ELISA in serum samples obtained after an overnight fasting.
Results.— Leptin levels were found significantly lower in migraineurs than controls (40.1 ± 21.2 ng/mL, 48.5 ± 24.5 ng/mL; P  < .05). Although body mass index did not differ between 2 groups, fat mass, and fat percentages were significantly lower in migraine patients (19.4 ± 8.8 kg, 26.0 ± 8.7 kg; P  < .001 and 28 ± 9%, 34 ± 5%; P  < .001, respectively).
Conclusion.— Migraine patients have low leptin levels and fat mass which may be related to the pathogenesis of migraine. The importance and impact of our findings on the prevalence, characteristics, and treatment of migraine needs to be investigated in further detailed studies.     

Improvement of fractional flow reserve and collateral flow by treatment with external counterpulsation (Art.Net.-2 Trial)

Background  Arteriogenesis (collateral artery growth) is nature's most efficient rescue mechanism to overcome the fatal consequences of arterial occlusion or stenosis. The goal of this trial was to investigate the effect of external counterpulsation (ECP) on coronary collateral artery growth.
Materials and methods  A total of 23 patients (age 61 ± 2·5 years) with stable coronary artery disease and at least one haemodynamic significant stenosis eligible for percutaneous coronary intervention were prospectively recruited into the two study groups in a 2 : 1 manner (ECP : control). One group (ECP group, n  = 16) underwent 35 1-h sessions of ECP in 7 weeks. In the control group ( n  = 7), the natural course of collateral circulation over 7 weeks was evaluated. All patients underwent a cardiac catheterization at baseline and after 7 weeks, with invasive measurements of the pressure-derived collateral flow index (CFIp, primary endpoint) and fractional flow reserve (FFR).
Results  In the ECP group, the CFIp (from 0·08 ± 0·01 to 0·15 ± 0·02; P  < 0·001) and FFR (from 0·68 ± 0·03 to 0·79 ± 0·03; P  = 0·001) improved significantly, while in the control group no change was observed. Only the ECP group showed a reduction of the Canadian Cardiovascular Society (CCS, P  = 0·008) and New York Heart Association (NYHA, P  < 0·001) classification.
Conclusion  In this study, we provide direct functional evidence for the stimulation of coronary arteriogenesis via ECP in patients with stable coronary artery disease. These data might open a novel noninvasive and preventive treatment avenue for patients with non-acute vascular stenotic disease.     

Anti-oxidized LDL antibody levels are reduced in women with hypertension

Background  Previous studies have suggested that hypertension may be associated with increased oxidized low-density lipoprotein (LDL). Increased in vitro oxidizability of LDL or elevated titers of anti-oxidized LDL antibodies have been shown in subjects with essential hypertension. However, the relationship between oxidized LDL and hypertension is equivocal. We examined the association between hypertension and levels of IgG anti-oxidized LDL antibodies in a group of women from the general population.
Materials and methods  The study included 619 women classified according to their blood pressure values. IgG anti-oxidized LDL antibodies were measured by enzyme-linked immunosorbent assay and the women were classified as being above or below the 50th percentile.
Results  Hypertension was present in 54·3% of the women. These women had significantly lower levels of IgG anti-oxidized LDL antibodies than the normotensive women (0·280 ± 0·117 vs. 0·336 ± 0·125, P  <   0·001). Both systolic and the diastolic blood pressures showed a significant negative correlation with the levels of IgG anti-oxidized LDL antibodies ( r  = −0·204, P  <   0·001; r  = −0·225, P  <   0·001, respectively). Women with IgG anti-oxidized LDL antibody levels above the 50th percentile had a lower prevalence of hypertension than those with IgG anti-oxidized LDL antibody levels below the 50th percentile (40·2% vs. 59·8%) ( P  <   0·001).
Conclusions  Women with hypertension had lower levels of IgG anti-oxidized LDL antibodies than normotensive women.     

Continuous monitoring of interface pressure distribution in intensive care patients for pressure ulcer prevention

Title.  Continuous monitoring of interface pressure distribution in intensive care patients for pressure ulcer prevention.
Aim.  This paper is a report of a study conducted to examine whether continuous interface pressure monitoring of postoperative patients in an intensive care unit is feasible in clinical practice.
Background.  The interface pressure between skin and surfaces is generally evaluated for pressure ulcer prevention. However, the intensity and duration of interface pressure necessary for pressure ulcer development remains unclear because the conventional interface pressure sensors are unsuitable for continuous monitoring in clinical settings.
Methods.  A total of 30 postoperative patients in an intensive care unit participated in this study in 2006–2007. A sensor was built into a thermoelastic polymer mattress. The whole-body interface pressure was recorded for up to 48 hours. Pressure ulcer development was observed during the morning bed-bath. For analysis, the intensity and duration of the maximal interface pressure was evaluated.
Findings.  The mean age of the study group was 62·0 ± 15·4 years. Two participants developed stage I pressure ulcer and blanchable redness at the sacrum. The longest duration of pressures greater than 100 mmHg were 487·0, 273·5 and 275·7 minutes in the pressure ulcer, blanchable redness and no redness groups respectively.
Conclusion.  Continuous monitoring of the intensity and duration of whole-body interface pressure using the KINOTEX sensor is feasible in intensive care patients.     

Evidence for intestinal oxidative stress in patients with obstructive jaundice

Background Obstructive jaundice results in failure of the intestinal barrier with consequent systemic endotoxemia associated with septic complications. We have recently shown that gut barrier failure in experimental obstructive jaundice is associated with high intestinal oxidative stress. This study was undertaken to investigate whether oxidative alterations occur in the intestinal mucosa of patients with obstructive jaundice. Patients and methods Fifteen patients with malignant biliary obstruction and no signs of cholangitis and 15 control patients were subjected to duodenal biopsy to assess intestinal oxidative stress, estimated by lipid peroxidation (malondialdehyde – MDA) and glutathione redox state [reduced glutathione (GSH), glutathione disulphide (GSSG) and GSH/GSSG ratio]. In addition, mucosal biopsies were examined histologically and intestinal mucosal protein content was determined biochemically as an index of intestinal trophic state. Results Patients with obstructive jaundice presented high levels of intestinal oxidative stress, with significantly increased lipid peroxidation (P < 0·001). Glutathione redox state was also suggestive of high intestinal oxidative stress in jaundiced patients, indicated by significantly decreased GSH (P = 0·001) and GSH/GSSG ratio (P = 0·006) and increased GSSG (P = 0·026). Histological examination showed a mild infiltration of the lamina propria by chronic inflammatory cells in obstructive jaundice, whereas duodenal architecture remained intact and epithelial continuity was retained. Duodenal mucosa was atrophic in jaundiced patients as indicated by a significant reduction of mucosal protein content compared with controls (P = 0·001). Among oxidative stress parameters, intestinal GSH exhibited a significant positive correlation with mucosal protein content (r = 0·588, P = 0·021). Conclusions Obstructive jaundice in humans induces intestinal oxidative stress, which may be a key factor contributing to intestinal barrier failure and the development of septic complications in this patient population.     

Medication information for Flemish inpatients with major depression: evaluation and construct validity of the Consumer Information Rating Form

Background and objectives:  Evaluating the quality of written medication information is a major topic of concern when patient pamphlets are developed. The objective of this study is to evaluate a patient pamphlet on selective serotonin reuptake inhibitors (SSRIs) by calculating Flesch-Douma readability scores and by applying the Consumer Information Rating Form (CIRF) to Flemish inpatients with major depression taking SSRIs.
Methods:  The pamphlet was evaluated by calculating Flesch-Douma readability scores. The study enrolled patients with major depression taking SSRIs. Patient received a SSRI pamphlet and completed a self-administered structured questionnaire consisting of the adapted CIRF and the Hospital Anxiety and Depression Scale. Construct validity was explored by means of factor analysis and Cronbach's alpha.
Results and discussion:  The Flesch-Douma readability scores showed that the pamphlet was easy to read, had much interest in the reader and was very popular. The sample of 96 patients consisted of doubtful/definite cases in terms of anxiety (10·65 ± 4·90) and doubtful cases in terms of depression (8·91 ± 5·23). Using the CIRF, patients assigned positive scores to comprehensibility, utility and design quality of the pamphlet. Factor analysis on the original CIRF confirmed the three original factors ( P  < 0·001). Cronbach's alpha of factors ranged from 0·69 to 0·83.
Conclusion:  The SSRI pamphlet is of good quality. Our study supported the construct validity of the CIRF to Flemish inpatients with major depression.     

High plasma levels of the soluble receptor for advanced glycation endproducts in patients with symptomatic carotid atherosclerosis

Background  Advanced glycation endproducts (AGEs), particularly carboxymethyl(lysine)-adducts (CML), exert part of their cellular effects by binding to a receptor, named receptor for AGEs (RAGE). The soluble form of this receptor (sRAGE) has been shown to have an athero-protective role. We hypothesized the existence of a relationship between the AGE–RAGE axis and the occurrence of symptoms related to carotid atherosclerosis in nondiabetic conditions.
Materials and methods  We evaluated plasma levels of CML and sRAGE (by ELISA), and tissue levels (tAGEs and tRAGE, semiquantitatively, by immunohistochemistry) in endarterectomy carotid plaque tissue in 29 nondiabetic patients. At the time of surgery, 10 patients were asymptomatic and 19 were symptomatic.
Results  Plasma levels of sRAGE were higher in symptomatic patients than in asymptomatic patients [median (interquartile range): 676 (394–858) pg mL⁻¹ vs. 347 (284–479) pg mL⁻¹, P  = 0·009]. In symptomatic patients, plasma levels of sRAGE correlated positively with CML ( r  = 0·60, P  < 0·01), C-reactive protein (CRP) ( r  = 0·618, P  < 0·01) and fibrinogen ( r  = 0·522, P <0·005), while in asymptomatic patients, no correlation was observed. Although tissue and plasma levels of AGEs and RAGE did not correlate between each other, tAGEs and tRAGE were also positively correlated only in symptomatic patients (χ² = 8·93, P  = 0·003).
Conclusions  Plasma levels of sRAGE are higher in symptomatic than asymptomatic carotid atherosclerosis. Higher levels of sRAGE in symptomatic patients may be markers of a higher degree of vascular inflammation in such patients.     

Outcome after device implantation in chronic heart failure is dependent on concomitant medical treatment

Background  Device implantation in chronic heart failure (CHF) for cardiac resynchronization therapy (CRT) with or without implantable cardioverter/defibrillator (ICD) is an established treatment option for symptomatic patients under medical baseline therapy. Although recommended, the need for optimization of medical therapy was never proven. As in 'the real world', medical therapy is not always up-titrated to the desirable dosages; this provides the opportunity to evaluate the impact of optimizing medical therapy in patients who had received a device therapy with proven effectiveness.
Materials and methods  This observational cohort study retrospectively assessed the 'real life'-effect of CRT compared with that of CRT/ICD therapy and the impact of concomitant pharmacotherapy on outcome. Outcome of patients with guideline recommended renin–angiotensin system inhibitor and ß-blocker dosages was compared with that of patients who failed to reach the desired dosages. Mean follow-up for the 205 CHF (95 CRT and 110 CRT/ICD) patients was 16·8 ± 12·4 months.
Results  In the total study cohort, 83 (41%) reached the combined primary endpoint of all-cause death or cardiac hospitalization [CRT group: 25 (26%), CRT/ICD group: 58 (52·7%), P  < 0·001]. Multiple cox regression analysis revealed non-optimized medical therapy at follow-up [HR = 2·080 (1·166–3·710), P  = 0·013] and CRT/ICD vs. CRT [HR = 2·504 (1·550–4·045), P  < 0·001] as significant predictors of the primary endpoint.
Conclusion  Our data stress the importance of professional monitoring and titration of pharmacotherapy not only in medically treated CHF patients but also in patients under device therapy by a heart failure unit or a specialized cardiologist.     

Effect of CYP3A5*3 genotype on serum carbamazepine concentrations at steady-state in Korean epileptic patients

Background and Objective:  Carbamazepine (CBZ) is metabolized mainly by the CYP3A family of enzymes, which includes CYP3A4 and CYP3A5. Several studies have suggested that the CYP3A5*3 genotype influences the pharmacokinetics of CYP3A substrates. The present study aimed to assess the effect of the CYP3A5*3 genotype on serum concentration of CBZ at the steady-state in Korean epileptic patients.
Method:  The serum concentrations of CBZ in 35 Korean epileptic patients were measured and their CYP3A5 genotype was determined. Fourteen patients were CYP3A5 expressors (two for CYP3A5*1/*1 and 12 for CYP3A5*1/*3 ) and 21 patients were CYP3A5 non-expressors ( CYP3A5*3/*3 ). Dose-normalized concentrations (mean ± SD) of CBZ were 9·9 ± 3·4 ng/mL/mg for CYP3A5 expressors and 13·1 ± 4·5 ng/mL/mg for CYP3A5 non-expressors ( P  = 0·032). The oral clearance of CBZ was significantly higher in CYP3A5 non-expressors than that of CYP3A5 expressors (0·056 ±0·017 L/h/kg vs. 0·040 ± 0·014 L/h/kg, P  = 0·004). The CYP3A5 genotype affected the CBZ concentrations in Korean epileptic patients and is a factor that may contribute to inter-individual variability in CBZ disposition in epileptic patients.     

Rapid and sensitive quantitation of morphine using HPLC with electrochemical detection

Background:  Morphine is one of the most widely used opioid analgesics for controlling pain in cancer and post-operative patients.
Objective:  This study aimed at finding a sensitive method for measuring morphine.
Method:  A one-step solid phase extraction was developed for extracting morphine from various samples, and morphine concentration was measured using a high-performance liquid chromatographic system with electrochemical detection. The sensitivity of the assay was 1·53 ng/mL with a recovery of 93·4% ± 0·01. The mean intra-assay and inter-assay test for three concentrations was 10·54 and 7·47, respectively. The assay showed no cross-reaction with a wide range of compounds.
Conclusion:  This method for morphine in small biological samples is easy, sensitive and reproducible with low cross-reactivity.     

Sirtuin gene expression in human mononuclear cells is modulated by caloric restriction

Background  Sirtuins may provide novel targets for treating some diseases associated with oxidative stress, such as obesity and its comorbidities. However, there are a few in vivo studies in humans about the potential role of sirtuins as therapeutic targets among obese patients undergoing caloric restriction. Therefore, the aim of this study was to assess if the gene expression of sirtuins is modulated in peripheral blood mononuclear cells (PBMC) by a hypocaloric diet devised to lose weight in humans.
Materials and methods  Gene expression of two sirtuins (SIRT1 and SIRT2) in the PBMC of obese subjects (32·3 ± 5·5 kg m⁻²) before and following an 8-week hypocaloric diet was investigated. NADH-coenzyme Q reductase (NDUFS2) and cytochrome  c  oxidase assembly protein (COX15) gene expression was selected together with plasma antioxidant power and nitric oxide as markers of antioxidant status. A quantitative real-time polymerase chain reaction approach was performed to assess the nutrigenomics outcome. Moreover, 2-keto[1-¹³C]isocaproate breath test (KICA-BT) parameters were evaluated to study mitochondrial oxidation in vivo .
Results  The intervention up-regulated the expression of both sirtuins, being inversely associated with total antioxidant capacity and directly related to nitric oxide, mitochondrial oxidation assessed by the KICA-BT and the expression of the mitochondrial proteins COX15 and NDUFS2.
Conclusion  SIRT1 and SIRT2 may serve as key regulators for some obesity comorbidities related to antioxidant status, while PBMC could be a model to study the effect of the sirtuin response in obesity therapy.