Evaluation of the Antioxidant Capacities and Cytotoxic Effects of Ten Parmeliaceae Lichen Species.

Parmeliaceae represents the largest and widespread family of lichens and includes species that attract much interest regarding pharmacological activities, due to their production of unique secondary metabolites. The current work aimed to investigate the in vitro antioxidant and cytotoxic activities of the methanol extracts of ten Parmeliaceae species, collected in different continents. Methanol extraction afforded high phenolic content in the extracts. The antioxidant activity displayed by lichens was evaluated through chemical assays, such as the ORAC (Oxygen Radical Absorbance Capacity) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities and the ferric reducing antioxidant power (FRAP). A moderately positive correlation was found between the phenolic content and the antioxidant properties for all the species: R: 0.7430 versus ORAC values, R: 0.7457 versus DPPH scavenging capacity, and R: 0.7056 versus FRAP reducing power. The methanol extract of Flavoparmelia euplecta exhibited the highest ORAC value, the extract of Myelochroa irrugans showed the maximum DPPH scavenging capacity, and Hypotrachyna cirrhata methanol extract demonstrated the highest reducing power. Further, the cytotoxic activity of the ten species was investigated on the human cancer cell lines HepG2 and MCF-7; Myelochroa irrugans exhibited the highest anticancer potential. The pharmacological activities shown here could be attributed to their phytochemical constituents.

Tratamiento rehabilitador en la incontinencia fecal / Rehabilitation treatment in fecal incontinence

Introducción. Podríamos definir la incontinencia fecal como la pérdida involuntaria de heces sólidas y líquidas, siempre que esta pérdida supone un problema higiénico o social en la persona que lo padece. La prevalencia de esta patología no está clara puesto que supone una importante afectación de la calidad de vida de los pacientes que la padecen, y muchas veces origina un encubrimiento de este problema. Material y método. Realizamos un estudio prospectivo, en el mismo han participado 24 pacientes diagnosticados de incontinencia fecal, todos ellos realizaron un programa de tratamiento que incluyó: normas educativas, ejercicios de fortalecimiento del suelo pélvico, y técnicas de biorretroalimentación con electroestimulación. Resultados. El 79,2% eran mujeres, la edad media fue de 60,8 años, el 20,8% de los pacientes habían tenido una neoplasia intestinal. En la pruebas complementarias encontramos en la ecografía que en un 25% de los casos había una rotura de alguno de los esfínteres anales, en un 70,8% un adelgazamiento de los esfínteres, en la manometría inicial encontramos, que la media de la presión máxima basal en mmHg fue de 36,37 mmHg (D.E. 13,13), y la media en la presión máxima en la contracción voluntaria fue de 82,25mmHg (D.E. 21,45) puntuación media inicial obtenida en la escala de Wexner fue de 15,79, y tras el tratamiento 8,16. Obtenemos diferencias estadísticamente significativas en todos los ítems de la escala de Wexner. Conclusiones. El tratamiento conservador combinado mejora la puntuación obtenida en la escala de Wexner en la incontinencia fecal moderada-severa, además de presentar mínimos efectos secundarios. Las pruebas complementarias son de utilidad para evaluar la incontinencia fecal, pero la evaluación clínica es fundamental para determinar la gravedad de esta patología, y la afectación que produce en la vida del paciente (AU)

Introduction. We can define fecal incontinence as the involuntary loss of solid and liquid stools, whenever this loss poses a hygiene or social problem for the person suffering it. Prevalence of this condition is not clear since it means an important affection in the quality of life of the patient suffering it and therefore, they often hide this problem. Material and methods. We have performed a prospective study in which 24 patients diagnosed of fecal incontinence participated. All of them underwent a treatment program that included: education guidelines, pelvic floor strengthening exercises and biofeedback with electrical stimulation. Results. A total of 79.2% were women, with mean age of 60.8 years, and 20.8% of the patients had suffered an intestinal neoplasm. In the complementary tests, the ultrasonography showed that 25% of the cases had a rupture of the anal sphincters. In 70.8%, there was thinning of the sphincters. In the initial manometry, we found that the measurement of maximum baseline pressure in mmHg was 36.37 mmHg (SD 13.13), and the mean maximum pressure involuntary contraction was 82.25mmHg (SD 21.45), initial mean score obtained on the Wexner scale was 15.79 and after treatment, 8.16. We obtained statistically significant differences in all of the items on the Wexner scale. Conclusion. Combined conservative treatment improves the score obtained on the Wexner scale in moderate to severe fecal incontinence and also shows minimum side effects. Complementary tests are useful to evaluate fecal incontinence, however clinical evaluation is fundamental to determine the severity of this condition and how it affects the patient's life (AU)

HPLC isolation of antioxidant constituents from Xanthoparmelia spp.

A chromatographic method is described for the purification and characterization of secondary lichen substances with biological activity. A simple reversed-phase high-performance liquid chromatography method with gradient elution has been developed that allows the determination and isolation of salazinic, usnic and stictic acids from lichen samples in a single run and the quantification of every acid in the tested extracts. The antioxidant activity of both the isolated compounds and the respective lichen belonging to Xanthoparmelia genus was determined by the Oxygen Radical Absorbance Capacity (ORAC) assay; their effect as free radical scavengers, effect on cell survival by the 3(4,5-dimethyltiazol-2-yl)-2,5-diphenyltetrazolium reduction assay and 2',7'-dichlorofluorescin diacetate method were tested on U373 MG human astrocytome cell line. Both lichens extracts and all isolated compounds protected U373 MG cells from hydrogen peroxide-induced damage, suggesting that they could act as antioxidant agents in those neurodegenerative disorders associated with oxidative damage, such as Alzheimer's disease and Parkinson's disease.

Whole blood glutathione peroxidase and erythrocyte superoxide dismutase activities, serum trace elements (Se, Cu, Zn) and cardiovascular risk factors in subjects from the city of Ponta Delgada, Island of San Miguel, The Azores Archipelago, Portugal.

Activities of whole blood glutathione peroxidase (GSH-Px) and erythrocyte superoxide dismutase (SOD) and serum levels of selenium (Se), copper (Cu) and zinc (Zn) were measured in 118 apparently healthy subjects aged 20-60 years from the city of Ponta Delgada, Island of San Miguel, The Azores Archipelago, Portugal. Data were analysed by age/gender, lipid profile and blood pressure as cardiovascular risk factors searching for their relevance when assessing reference values for antioxidant biomarkers. GSH-Px was in the same range, but SOD was significantly lower than in other Portuguese populations. Neither activity differed with gender. GSH-Px activity increased with age, namely in normolipidemic men versus the hyperlipidemic group in which a decrease was observed. This suggests a progressive impairment of GSH-Px with age caused by an enhanced production of oxidant species in hyperlipidemia. GSH-Px was 30% lower in male hypertensives versus normotensives. SOD activity did not relate to age or blood pressure but was 17% higher in the hyperlipidemic men versus the normolipidemic group, suggesting a better antioxidant protection by SOD than by GSH-Px in hyperlipidemia and hypertension. Se was higher in men versus women, particularly in the older subjects, and partly related to hyperlipidemia. Zn levels showed a similar dependency on gender, not related to age or lipid profile. Cu levels were much higher in women than in men in all age or lipid profile classes and decreased in hyperlipidemia. They were lowered with age in both genders, particularly in normolipidemic women. The present research therefore suggests that hyperlipidemia and hypertension do affect antioxidant status and should be considered when assessing antioxidant biomarkers in blood.

Trace element status (Se, Cu, Zn) in healthy Portuguese subjects of Lisbon population: a reference study.

Serum levels of selenium, copper, and zinc were systematically determined in healthy subjects of the Lisbon population. The sample consisted of 183 blood donors of both genders who were divided into two age groups: 20-44 and 45-70 yr of age; relationships with gender, age, the lipid profile, and tobacco consumption were investigated. In the female group, the intake of oral contraceptives and pregnancy were considered for the youngest women, and hormonal replacement therapy (HRT) was taken into account for the oldest ones. Serum concentrations of these elements were in the same range as those found for populations of other European countries. Differences between genders were observed for the three elements studied, with serum selenium and zinc concentrations higher in men and copper levels higher in women. Age-dependent differences were found for selenium: The oldest subjects (regardless gender) presented the highest concentrations of selenium in serum as further demonstrated by the positive correlation with age. In both pregnant and contraceptive-using women, copper was greatly increased, confirming the influence of estrogen status and/or oral contraceptive intake on increased serum copper levels. However, in postmenopausal women, HRT did not significantly affect serum copper levels. Selenium, copper, and zinc status were not different between normolipidemic and hyperlipidemic subjects for the same gender and age range, but selenium levels tended to increase with hyperlipidemia when considering the whole group of subjects. With respect to the lifestyle, higher serum zinc levels were found in tobacco-consuming men. Albumin serum levels were similar for all considered subgroups, except for the pregnant women, for whom a decrease in this parameter was observed. The present study allowed one to obtain reference values for this healthy group of population, which will serve for a comparative study with groups having pathological conditions, such as cardiovascular disease.

Systemic markers of the redox balance in chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is highly prevalent and its pathogenesis is still not completely clarified. Clinically stable patients (n=21) and healthy subjects (n=24) were studied for blood markers of oxidative injury and antioxidant status. The plasma concentration of protein carbonyls was significantly increased in COPD patients, both ex-smokers (0.76 +/- 0.28 nmol mg(-1)) and smokers (0.99 +/- 020 nmol mg(-1)) versus controls (0.49 +/- 0.14 nmol mg(-1)) . The concentration of total thiols was slightly enhanced in plasma of the COPD patients (ex-smokers 492 +/- 23 micromol 1(-1) and smokers 505 +/- 36 micromol 1(-1) versus controls 450 +/- 67 micromol 1(-1); p < 0.05). The activity of the antioxidant enzyme superoxide dismutase was increased in erythrocytes (activity in U g(-1) haemoglobin; ex-smokers 4460 +/- 763 and smokers 4114+/- 1060 versus 3015 +/- 851 in controls; p > 0.01), while glutathione peroxidase activity was decreased in total blood (activity in U g(-1) haemoglobin: ex-smokers 27 +/- 9 and smokers 23 +/- 9 versus 47 +/- 25; p < 0.01). Lower levels of selenium in plasma were also found for COPD patients (concentration in mg 1(-1): ex-smokers 0.030 +/- 0.019 and smokers 0.032 +/- 0.024 versus 0.058 +/- 0.023 in controls; p < 0.01), being more evident in those with very low levels of arterial oxygen pressure. In addition, the levels of potassium and rubidium were increased in blood cells of the patient group. All these changes might reflect oxidant damage and an altered electrolytic homeostasis, and can be interpreted as markers of COPD rather than as indicators of smoking habits.

Hepatic elemental contents and antioxidant enzyme activities in Algerian mice (Mus spretus) inhabiting a mine area in central Portugal.

In this study the effects of heavy metals (manganese, iron, copper, zinc) and selenium exposure on the hepatic activity of antioxidant enzymes, superoxide dismutase (SOD) and glutathione S-transferases (GST), were appraised on a seasonal basis in Algerian mice (Mus spretus) inhabiting an active copper mine area. A reference population of the same species was considered for comparative purposes. Different patterns of seasonal variation were found in both populations for the manganese, iron and selenium hepatic concentrations and SOD activity. When the two populations were compared, iron and selenium concentrations were enhanced in mice from the polluted area. In addition, SOD activity was significantly decreased in summer in exposed mice, but no other significant changes in SOD and GST activities between sites throughout the year were recorded. However, when seasonal data within each group of mice were pooled, significant differences were found between sites for the average concentrations of manganese, iron and selenium, which are higher in the polluted site. In addition, significant differences were obtained for the average values of SOD and of GST activities, due to simultaneously higher GST values and slightly lower SOD values in the polluted site. The population from the reference site was more homogeneous for all parameters measured than the population from the polluted area. These results, in particular the higher variability in data collected from mice exposed to heavy metals and selenium, combined with the negative associations between biochemical markers and heavy metals, may suggest, despite the good adaptability of the mice to their habitat, biochemical stress due to diminished environmental quality.

Comparison of whole-blood glutathione peroxidase activity, levels of serum selenium, and lipid peroxidation in subjects from the fishing and rural communities of "Rabo de Peixe" village, San Miguel Island, the Azores' Archipelago, Portugal.

The activity of glutathione peroxidase (GSH-Px), serum selenium (Se), and thiobarbituric acid reactive substances (TBARS) were measured in the whole blood of 148 healthy adults aged 20-60 yr from the fishing and rural communities of "Rabo de Peixe," The Azores, Portugal. The subjects did not live in the same household and had different socioeconomic profiles and dietary habits. The serum lipid profile and selected life habits were also considered in this study. No significant differences in the activity of GSH-Px were found in the interpopulation or intrapopulation analyses, classified by age or lipid profile. An age-dependent GSH-Px increase was noted in the younger male (M) subgroups (20-39 yr). The Se levels were higher in fishers (f) of both genders (M, F) than in subjects living in the rural (r) environment: 110+/-25 microg/L (f, M), 89+/-20 microg/L (f, F), 88+/-22 microg/L (r, M) and 80+/-17 microg/L (r, F). In the fishers, but not in the rural population, Se was higher in the males, but it did not show significant variation with age. The levels of TBARS were lower in the f than in the r male group. The Se level was lower and TBARS higher in the hyperlipemic women in the f group, compared to the corresponding controls. Our results suggest that the fishers (mainly men) show a better antioxidant status than that of their rural counterparts, due to differences in dietary habits between the study populations and between genders.

Response of antioxidant enzymes in freshwater fish populations (Leuciscus alburnoides complex) to inorganic pollutants exposure.

Evidence is accumulating indicating the importance of antioxidant enzyme activity measurements in eco-toxicological studies, as they may constitute markers for exposure to a large variety of pollutants. Variation of antioxidant enzymes, superoxide dismutase (SOD) and glutathione S-transferases (GST) and the effect of heavy metals and selenium exposure on these enzymes were investigated in the livers of Iberian endemic minnows (Leuciscus alburnoides complex) captured in a copper (Cu) mining area. Higher hepatic levels of copper and selenium were always observed in fish captured at the polluted site relative to the reference area population, reflecting the environmental monitoring results. A seasonal fluctuation in zinc and selenium levels for both populations was also observed which could be related to gonad maturation. The activity of SOD did not show significant regional alterations, but a seasonal variation occurred presumably associated with the Leuciscus life cycle. The GST activity was higher in the fish population from the polluted area (except in spring) and GST variability was associated with selenium and copper levels when both regions were compared. The increased GST activity was probably a metabolic adaptation to the continuous exposure to higher levels of those elements.

The effect of age and selenium on some biochemical parameters in rat liver.

Two age groups, 3 and 15 mo, were used to investigate whether age-associated changes in some parameters related to lipid peroxidation occur in the liver of male Wistar rats and to observe possible effects of dietary selenium supplementation (0.25 and 0.50 ppm) for 12 mo on the same parameters. At these experimental conditions, the most important observation was that peroxidation did not change by aging, at least until 15 mo of age. In addition, the activity of Se-dependent glutathione peroxidase (GSH-Px, EC 1.11.1.9) was higher in the liver of the older animals. It is suggested that the enzyme could have a role in the unchanged hepatic peroxidation observed in aged male rats. On the other hand, an effect of dietary selenium supplementation on those parameters was not observed, probably because the selenium levels were still at an adequate plateau.

Effect of selenium supplementation on polyunsaturated fatty acids in rats.

The purpose of this work was to study plasma, adipose tissue, and liver fatty acids percentages of Wistar rats that drank water supplemented with several levels of sodium selenite for 1, 3, and 6 mo. In a general way, percentages of saturated, monounsaturated, and polyunsaturated fatty acids of supplemented groups were not different from those obtained with nontreated animals in the analyzed tissues. However, in rats supplemented with 0.5 ppm Se, mainly in adipose tissue, a polyunsaturated fatty acids increase (p < 0.005) was observed for all times of treatment. This could suggest that 0.5 ppm Se supplement probably exercises a protective role on polyunsaturated fatty acids in that tissue. Supplements of 6.0, 15.0, and 54.0 ppm Se did not change unsaturation levels of fatty acids in the analyzed tissues.

Effect of selenium supplementation on some blood biochemical parameters in male rats.

The long-term effect of selenium supplementation on blood glutathione peroxidase (GSH-Px) activity and plasma TBARS' production (as an index of peroxidation) was evaluated in 15-mo-old male rats fed a diet supplemented with 0.25 or 0.50 ppm selenium, for 12 mo. A group of nonsupplemented age-matched rats was the control. In addition, triglycerides, phospholipids, total and free cholesterol, HDL-cholesterol, and HDL-phospholipids levels were measured in plasma. Plasma testosterone levels were also determined in order to control the aging process in these animals. The GSH-Px activity and the peroxidation level were unchanged in all the groups. However, concerning the lipid parameters, a decrease in triglycerides concentration was observed in both treated groups (p < 0.05). Therefore, in these experimental conditions, despite no observed changes in parameters related to lipid peroxidation, selenium seems to be involved with triglycerides metabolism, eventually improving the triglycerides status of aged animals.

Selenium and lipid parameters in plasma of Portuguese subjects.

To obtain further insight into the role of selenium in the development of atherosclerosis, plasma selenium and lipid parameters were determined in 126 Portuguese subjects living in the region of Lisbon, 60 women and 66 men, 20-60 years old, divided up in to three groups according to their plasma lipid profile: Group I consisted of normolipidemic subjects having plasma triglycerides and total cholesterol < 200mg/dL; Group II was composed of hypercholesteremic subjects with total cholesterol > 200mg/dL and plasma triglycerides < 200mg/dL; Group III was composed of hypercholesteremic and hypertriglyceremic subjects having total cholesterol and plasma triglycerides > 200mg/dL. Respective values for mean plasma selenium (+/- SD) in the groups were: 93 +/- 18 micrograms/L, 90 +/- 17 micrograms/L and 96 +/- 18 micrograms/L. A weak significant difference in plasma selenium between women and men was, however, observed in Group I (87 +/- 20 micrograms/L vs. 100 +/- 11 micrograms/L, p < 0.05). Regression analysis showed no significant relationship between plasma selenium and lipid parameters, except in the most hyperlipidemic women of Group III, where a weak correlation between plasma selenium and the HDL-cholesterol/total cholesterol ratio (r = 0.80, p < 0.05) was observed. The present study suggests that the relationship between selenium and HDL-cholesterol has to be further examined, taking into account nutritional, metabolic and genetic factors.

Plasma and liver selenium levels in the rat during supplementation with 0.5, 2, 6, and 15 ppm selenium in drinking water.

Plasma and liver selenium of Wistar rats were determined after 1, 3, and 6 mo supplementation with 0.5, 2, 6, or 15 ppm selenium as sodium selenite in drinking water. Plasma selenium was not different from control values at additional intake of 0.5 ppm but increased above usual levels at higher intakes. A highly significant correlation was observed between the total quantity of selenium ingested and plasma selenium after 1 mo treatment (r = 0.99, p < 0.01), but was less pronounced after 3 and 6 mo (0.94, p < 0.05, and 0.78, p < 0.05, respectively). The decrease in plasma selenium with time of treatment was more pronounced at higher intakes. There was also a highly significant correlation between total selenium intake and liver selenium concentration (r = 0.99, p < 0.01) after 1 mo of treatment, but this time liver selenium did not change with time, and the correlation remained highly significant throughout the investigation. Liver selenium therefore appears as a more sensitive and more representative measure of selenium intake than plasma selenium. Most supplements did not affect body weight and survival of animals, except when the diet was supplemented with 15 ppm for 6 mo; however, alterations in biochemical parameters concerning lipid status and hepatic function were observed at levels above 2.0 ppm.