[ALPHA-1-ANTITRYPSIN ACTIVITY IN PATIENTS HOSPITALIZED FOR CELLULITIS].

INTRODUCTION: Deficiency or impaired activity of alpha-1-antitrypsin (AAT), which neutralizes multiple proteolytic enzymes, such as collagenases and elastases may result in significant tissue autodigestion. Hence, AAT may have a role in the healing process in chronic and acute inflammation including skin infection, such as cellulitis. AIM: The aim of this study was to evaluate the role of AAT activity and inflammatory markers in patients with cellulitis. METHODS: The study included eleven consecutive patients (6 males and 5 females, mean age 68.5 ± 4.5 years) who were hospitalized for cellulitis between 09/2009-02/2010. We analyzed tests results for C reactive protein (CRP), AAT level and activity that were obtained on admission (T1), 2 days after admission (T2) and 2 weeks after admission (T3). RESULTS: AAT levels were found to be within the normal range. AAT activity values were found to be within or above the normal range. The highest activity values were measured after 2 days of treatment and the lowest values were measured after 2 weeks of treatment. CRP values were highest on admission and lowest, as expected, after the end of treatment 2 weeks later. AAT activity values were significantly lower statistically in patients with unresolved cellulitis 2 weeks after treatment began. SUMMARY: AAT activity was significantly lower statistically in patients who suffered from slow resolving cellulitis 14 days after hospitalization. This possibly suggests a role AAT activity may have in the inflammation cascade in patients with cellulitis. Further studies are needed to evaluate the role of AAT activity in the inflammatory process.

The Genetics of Glucose-6-Phosphate-Dehydrogenase (G6PD) and Uridine Diphosphate Glucuronosyl Transferase 1A1 (UGT1A1) Promoter Gene Polymorphism in Relation to Quantitative Biochemical G6PD Activity Measurement and Neonatal Hyperbilirubinemia.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency and polymorphism in uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1) were associated with significant neonatal hyperbilirubinemia (NHB) and increased risk for kernicterus. However, quantitative screening tests for G6PD enzyme activity proved unsatisfactory in estimating the risk for significant NHB, especially in heterozygous females that could present phenotype overlap between normal homozygotes, heterozygotes, and deficient homozygotes, resulting in a continuum of intermediate G6PD activity. OBJECTIVE: To examine the association of genotype and phenotype in newborns with decreased G6PD activity and its relation to NHB. STUDY DESIGN: Quantitative G6PD enzyme activities were measured on umbilical cord blood samples. After accepting parental consent, samples were analyzed for G6PD mutations and UGT1A1 gene polymorphisms (number of TA repeats in the UGT1A1 promoter). The associations to quantitative G6PD activity and bilirubin levels were assessed. RESULTS: 28 females and 27 males were studied. The Mediterranean mutation (NM_001360016.2(G6PD): c.563C>T (p.Ser188Phe)) was responsible for most cases of G6PD deficiency (20 hemizygous males, 3 homozygous and 16 heterozygous females). The association between this mutation, decreased G6PD activity and higher bilirubin levels was confirmed. Heterozygosity to 6/7 TA repeats in the UGT1A1 promoter was associated with increased NHB, especially in female newborns with G6PD deficiency. However, it seems that the interaction between G6PD deficiency, UGT1A1 promoter polymorphism, and NHB is more complex, possibly involving other genetic interactions, not yet described. Despite genotyping females with G6PD deficiency, the overlap between the upper range of borderline and the lower range of normal G6PD activity could not be resolved. CONCLUSIONS: The results of this study highlight the possibility for future implementation of molecular genetic screening to identify infants at risk for significant NHB, especially UGT1A1 polymorphism in heterozygous females with borderline G6PD deficiency. However, further studies are needed before such screening could be applicable to daily practice.

Umbilical Cord and Neonatal Transthyretin and Their Relationship to Growth and Nutrition in Preterm Infants.

BACKGROUND: Transthyretin (TTR), also known as prealbumin, has been suggested as an indicator of protein and nutritional status. OBJECTIVE: The aim of this study was to examine the maternal and umbilical cord (UC) TTR in relation to intrauterine growth, and the serum TTR of preterm infants in relation to nutritional status and growth. METHODS: After application of exclusion criteria, 49 preterm infants (mean gestational age and birth-weight 32.9±2.9 weeks and 1822±556 g) were included in the study. Transthyretin was sampled at birth and on days 14, 28, and at discharge with growth parameters and nutritional laboratory test results. RESULTS: Mean UC and maternal TTR were positively correlated (8.5±2.4 mg/dL and 20.4±7.0 mg/dL, r=0.31, P=0.07). Umbilical cord TTR was neither an index of maturity nor of intrauterine growth. Umbilical cord TTR was higher in females (9.4±2.6 versus 7.6±1.8 mg/dL, P=0.015). Maternal TTR was lower in twin pregnancies (16.8±4.9 versus 22.5±7.3 mg/dL, P=0.007). Although TTR levels gradually increased over time in correlation with post-menstrual and chronological ages (r=0.24, P=0.011 and r=0.40, P<0.001, respectively), there was no correlation to weight gain (r=0.10, P=0.41), nutritional status, protein intake, or nutritional laboratory test results. The only significant correlations were between TTR and glucose and triglycerides levels (r=0.51, P<0.001 for both). CONCLUSIONS: Although TTR levels increased over time, we could not demonstrate significant correlations between TTR and indices of the nutritional status in preterm infants at birth or during the neonatal course.

Exercise training modulates cytokines activity in coronary heart disease patients.

BACKGROUND: Physical activity may lower the risk for coronary artery disease (CAD) by mitigating inflammation, which plays a key role in the pathophysiology of atherosclerosis. The purpose of this study was to determine the effect of aerobic exercise training on levels of pro- and anti-inflammatory cytokines, IL-1, IL-6, IL-10, INF-gamma, and C-reactive protein (CRP), in CAD patients participating in a cardiac rehabilitation program. METHODS AND RESULTS: Twenty-eight patients, age 64+/-7.1 years, participated in a 12-week aerobic exercise training program at 70-80% of individual maximal heart rate. Training resulted in a significant reduction of all pro-inflammatory cytokines, CRP from 7.5+/-4.2 to 3.9+/-3.5 mg/l, p<0.001, IL-1, 0.33+/-0.23 to 0.51+/-0.12 pg/ml, p=0.014, IL-6, 2.50+/-1.50 to 1.44+/-0.57 pg/ml, p=0.002, INF-gamma, 18.63+/-3.31 to 16.77+/-2.49 pg/ml, p<0.001, as well as a significant increase in the anti-inflammatory, cytokine IL-10, from 1.61+/-1.40 to 2.29+/-2.01 pg/ml, p=0.008. Baseline CRP levels were 36% (p=0.006) higher among diabetes mellitus patients and training was associated with a 40.5% CRP reduction in these patients compared to 19% reduction in non-diabetics, p<0.01. At baseline 72% of patients were in a high risk category (CRP>3 mg/l), 28% in an intermediate (CRP=1-3 mg/l), with none in a low risk category (<1 mg/l). Following exercise training, 11% were in the low risk, 50% in the intermediate and 39% in the high risk category, indicating 46% reduction in the number of subjects in the high risk category. CONCLUSIONS: Aerobic exercise training in CAD patients is an effective mean in inducing reduction in CRP, IL-1, IL-6, INF-gamma levels, and increase in IL-10, thus, possibly improving coronary risk profile.

Adenosine deaminase activity in patients with Crohn's disease: distinction between active and nonactive disease.

BACKGROUND: Crohn's disease (CD) is a common chronic inflammatory bowel disease. During the disease a cascade of immunologic events occur including mucosal influx of inflammatory cells like neutrophils. Adenosine deaminase (ADA) is important in inflammatory responses and serves as a marker of activated leukocytes. MATERIALS AND METHODS: In this study, we investigated the activity of total ADA (tADA) and its isoenzymes, ADA1 and ADA2, in serum and neutrophils derived from 20 active patients with CD, 20 patients in remission, as well as in 15 healthy controls. RESULTS: Patients with active disease had significantly (P<0.001) higher levels of tADA in serum (22.9±4.9 U/l) than patients in remission or healthy controls (14.0±3.4 U/l and 13.2±2.4 U/l respectively). ADA2, the main isoenzyme in the serum was higher in active patients by 60% as compared with patients in remission and healthy controls (19.7±1.9 U/l, 12.3±1.2 U/l, and 12.2±0.9 U/l respectively). We did not find a significant difference in these parameters between healthy controls and stable patients. There was a positive correlation (R=0.516) between tADA activity and C-reactive protein levels in patients with CD. Enhanced activity in tADA was also detected in neutrophils that were obtained from all patients with CD as compared with healthy controls (15.3±2.9 U/g, 14.1±2.3 U/g, and 9.4±2.9 U/g protein, respectively). This is mainly due to a significant increment (up to 51%) in ADA1 activity, the main isoenzyme in the neutrophils (84% out of the tADA). The cause of this increment remains to be elucidated. CONCLUSION: The results obtained in this study demonstrated elevated levels of tADA and ADA2 in patients with active disease. As the patient improves and becomes clinically stable these levels decrease, approaching normal values. tADA and ADA2 can be used as markers of inflammation, and provide a supportive indicator of CD activity.

Oxidative stress, inflammation and neutrophil superoxide release in patients with Crohn's disease: distinction between active and non-active disease.

Increased oxidative stress has been previously demonstrated in patients with Crohn's disease (CD). However, to date, this parameter has not been assessed in a comparative study of patients in prolonged remission and those with the active disease. We report here our study of lipid peroxidation, antioxidant and inflammation status in serum derived from 16 active CD patients, 27 clinically stable patients, and 15 healthy controls. Results The extent of lipid peroxidation was higher in CD patients than in the healthy controls, while the levels of lipid peroxides (PD) and of thiobarbituric acid-reactive substances (TBARS) were significantly (P < 0.01) higher in serum obtained from patients with active CD (22 and 30%, respectively) than in that obtained from patients in remission. An analysis of the antioxidant status revealed that the beta-carotene levels in sera derived from all CD patients - patients with active or stable CD (49.4 +/- 15 and 95.6 +/- 25 mg% beta-carotene, respectively) - were higher than that in the controls (145 +/- 40 mg%). Serum activity of glutathione peroxidase (GSH-Px) was significantly (P < 0.001) higher (by 31%) in the patients with active CD than in the control group. There was no significant difference in GSH-Px activity between patients in remission and the controls. In terms of the inflammatory status, we found significantly (P < 0.01) higher levels of C-reactive proteins (CRP) and of tumor necrosis factor alpha (TNFalpha) in patients with active CD than in CD patients in remission. There was a significant correlation between those parameters and the extent of lipid oxidation. Neutrophils, which are a potential source of oxygen-free radicals, were activated by incubation with phorbol myristate acetate (PMA). Superoxide and lysozyme release were significantly reduced in neutrophils derived from patients with active CD (by 25 and 28%, respectively) in comparison to the control group. However, stimulated neutrophils from stable patients demonstrated only a minimally non-significant lower release of superoxide and lysozyme compared to the controls. Conclusion The results obtained in this study demonstrate an enhanced inflammatory and oxidative stress and a decreased antioxidant status in patients with active CD. As the patients improved and became clinically stable, the oxidative parameters decreased, approaching normal values. As neutrophil activation was also lower in patients with active disease, neutrophil activation may represent a possible defense mechanism of the body against tissue injury.

Anthropic arguments and the cosmological constant, with and without the assumption of typicality.

We reexamine claims that anthropic arguments provide an explanation for the observed smallness of the cosmological constant, and we argue that correlations between the cosmological constant value and the existence of life could at best be demonstrated only under restrictive assumptions. Causal effects are more subtle to uncover. The assumption of our typicality is crucial to such arguments.

Enhanced oxidative stress and leucocyte activation in neoplastic tissues of the colon.

Excess of intracellular reactive oxygen species results in an environment that may modulate gene expression, or damage cellular molecules. These events are assumed to contribute to the process of carcinogenesis. In the present study, we measured the extent of lipid peroxidation and antioxidative status in colonic tumors and normal colonic mucosa obtained from 25 patients with colorectal carcinoma. Levels of lipid peroxides (PD) and of thiobarbituric acid reactive substances (TBARS) were significantly increased, by 54 and 59%, respectively, in tissue specimens obtained from the colonic tumor as compared with normal colonic mucosa (PD, 2.78+/-0.31 versus 1.81+/-0.29 nmol/mg tissue, TBARS, 0.86+/-0.1 versus 0.54+/-0.08 nmol/mg tissue). Activities of the antioxidant enzymes catalase and glutathione peroxidase (GPx) were also higher (by 67 and 29%, respectively) than in normal mucosa, probably in response to the increased free radical stress occurring in cancerous tissues. Myeloperoxidase (MPO) and adenosine deaminase (ADA) are markers of activated leukocytes and are related to the production of oxygen free radicals by these cells. Their activities were significantly elevated in the neoplastic tissue as compared to the normal tissue (MPO, 7.4+/-1.5 versus 4.1+/-0.95 U/mg tissue, ADA, 4.17+/-0.65 versus 2.99+/-0.80 U/g tissue), suggesting a possible involvement of activated leukocytes in the enhanced oxidative stress in the cancerous tissue. Our results demonstrate an enhanced oxidative stress in the neoplastic tissue. Leukocyte activation was also higher in the carcinogenic tissue, indicating a possible contribution of these cells to a further oxidative stress-derived tissue injury. These observations add to previous studies and may encourage therapeutic trials with antioxidants as a means of preventing colorectal cancer and preventing further tissue injury in the neoplastic tissue and its surroundings.

The early anti-oxidant effect of carvedilol predicts the clinical course in congestive heart failure patients.

BACKGROUND: Carvedilol, a beta1 and beta2 as well as an alpha1 adrenoreceptor antagonist with multiple hemodynamic, anti-ischemic and anti-oxidant properties, is widely accepted for the treatment of hypertension and congestive heart failure (CHF). It has been shown to improve morbidity and mortality in CHF. OBJECTIVES: To assess whether the anti-oxidant effect of carvedilol has an impact on the clinical course in post-myocardial infarction (MI) CHF. METHODS: Thirty-nine recent MI patients, aged 60.5 +/- 7 years, New York Heart Association functional class (FC) II-III, and left ventricular ejection fraction (LVEF) 29 +/- 3.8%, underwent oxygen free radical (OFR) assessment using the thiobarbituric acid reactive substances, thermochemoluminescence and conjugated dienes methods. OFR was determined at baseline, 1, 3, 12, and 24 h after 3.125, 6.25 and 12.5 mg carvedilol, and after 6 months of treatment. Brain natriuretic peptide (BNP), LVEF, FC change, 6-min walk test (6MW) and quality of life scores were evaluated before and after 6 months. RESULTS: Two patterns of OFR activity were found. In 29 patients (group 1) a significant and consistent reduction in OFR following administration of each dose of carvedilol was found, significantly correlating with each of the outcome parameters. In ten patients (group 2), no change in OFR was found, nor in any of the other outcomes. At 6 months, FC improved in 23 patients from group 1 (79.3%) and only in one (10%) from group 2 (P<0.01). 6MW increased by more than 10% in group 1 with no change in group 2 (P<0.05). BNP decreased from 397 +/- 36 pg/ml to 171 +/- 15.9 pg/ml (P<0.01) in group 1 compared to 381 +/- 32.5 pg/ml and 405 +/- 36 pg/ml, respectively (P=not significant) in group 2. One year hospital admissions and death rate were significantly higher in group 2. CONCLUSIONS: The early anti-oxidative effect of carvedilol correlates well with the clinical course and probably predicts it.

Paraoxonase activity following exercise-based cardiac rehabilitation program.

PURPOSE: The effect of gender on paraoxonase activity was determined in 37 ischemic heart disease patients who underwent a 12-week aerobic exercise training program. METHODS: Paraoxonase activity was measured by its arylesterase activity (spectrophotometrically, at 250 degrees C, wavelength 270 nm). RESULTS: A 16.7% increase in paraoxonase activity was found following the 12-week exercise program. In addition, there was a significant gender effect with higher mean paraoxonase levels among women during both preexercise (20.8%) and postexercise (24.2%) testing. CONCLUSIONS: Aerobic exercise training was found to be an effective means in inducing plasma levels elevation of the antioxidative, antiatherogenic paraoxonase in patients with coronary artery disease, and particularly in female patients.

Modifications and oxidation of lipids and proteins in human serum detected by thermochemiluminescence.

Detection of electronically excited species (EES) in body fluids may constitute an important diagnostic tool in various pathologies. Examples of such products are triplet excited carbonyls (TEC), which can be a source for photon emission in the 400-550 nm range. The aim of the present study was to determine the actual contribution of lipid and protein components (protein carbonyls) to photon emission generated by thermochemiluminescence (TCL) during the heating of biological fluids. In this study, a new TCL Photometer device, designed by Lumitest Ltd, Israel, was used. Samples were heated to a constant temperature of 80 +/- 0.5 degrees C for 280 s and photon emission was measured at several time points. In order to compare the results of TCL measurements to conventional methods of detecting lipid and protein oxidation, each examined sample was also heated in a waterbath at 80 degrees C for 10-280 s. Lipid and protein oxidation were subsequently measured using conventional methods. The TCL of four polyunsaturated fatty acids (PUFA) with three to six double bonds was measured. The elevation of the PUFA TCL amplitude correlated with the increase in the number of double bonds of PUFA. A correlation between the increase in TCL intensity and protein carbonyl generation in bovine serum albumin (BSA) was also observed. In the venous blood serum, our study showed that an increase of TCL intensity during heating reflected the cleavage of TEC of lipid origin. Our study suggests that biological molecules such as proteins, lipids and other molecules, which may become unstable during heating, are capable of generating EES. We demonstrated that a TCL curve can be used as a kinetic model for measuring oxidative processes, which reflects modifications of different molecules involved in the oxidative stress phenomena.