Evaluation of the effectiveness of three physiotherapeutic treatments for subacromial impingement syndrome: a randomised clinical trial.

OBJECTIVE: To determine whether dexketoprofen administered by phonophoresis or iontophoresis is more effective for the treatment of subacromial impingement syndrome (SIS) than conventional ultrasound therapy. DESIGN: Randomised clinical trial. SETTING: University hospital. PARTICIPANTS: Ninety-nine participants with SIS without a complete tear of the rotator cuff were assigned at random to three intervention groups. INTERVENTION GROUPS: Participants received ultrasound (n=32), phonophoresis with dexketoprofen (50mg/session) (n=33) or iontophoresis with dexketoprofen (50mg/session) (n=34). All participants completed 20 treatment sessions plus exercise therapy and cryotherapy. OUTCOME MEASURES: A visual analogue scale (VAS), the Constant-Murley Scale (CMS) and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire were administered pre-treatment (baseline), post-treatment and 1 month post-treatment. RESULTS: At baseline, there were no differences between the groups. Post-treatment, VAS score improved by -1.2 points and CMS score improved by 8.9 points in the ultrasound group compared with the iontophoresis group [95% confidence interval (CI) -0.2 to -2.2 and 95% CI 17.0 to 0.7, respectively]. CMS score improved by 7.1 points in the phonophoresis group compared with the iontophoresis group (95% CI 14.8 to -0.7). At 1 month post-treatment, no significant differences were detected between the groups. VAS, CMS and DASH scores of all groups improved post-treatment and at 1 month post-treatment. CONCLUSION: Ultrasound, iontophoresis with dexketoprofen and phonophoresis with dexketoprofen can improve pain, shoulder function, and physical functioning and symptoms in the upper limb in patients with SIS without a complete tear of the rotator cuff. CLINICAL TRIALS. GOV REGISTRATION NUMBER: NCT01748188.

Parameters related to oxygen free radicals in human skin: a study comparing healthy epidermis and skin cancer tissue.

In vitro studies with tumor cells have demonstrated that oxygen free radicals are involved in the development of skin cancers and that variations in the body's defense mechanisms can modify the course of the disease. To assess the validity of this hypothesis in spontaneous tumors, we determined glutathione S-transferase, superoxide dismutase, reduced and oxidized glutathione, and thiobarbituric acid reactive substances in healthy whole skin (n = 95), dermis (n = 73), and epidermis (n = 69). The values were compared with those obtained in three types of skin cancer: basal cell carcinoma (n = 16), squamous cell carcinoma (n = 6), and melanoma (n = 33). In healthy skin, glutathione S-transferase, superoxide dismutase, reduced glutathione, and oxidized glutathione were higher in epidermis than in dermis, whereas thiobarbituric acid reactive substances were higher in dermis than in epidermis; whole skin had intermediate values. These results suggest that there is an induction of some anti-oxygen free radicals mechanisms in epidermis as a result of increased oxygen free radicals production. Glutathione S-transferase and thiobarbituric acid reactive substances were higher in all types of tumor than in healthy epidermis but oxidized glutathione was lower. Reduced glutathione and superoxide dismutase activity were lower in basal cell carcinoma and squamous cell carcinoma samples. Glutathione S-transferase increased, whereas superoxide dismutase and thiobarbituric acid reactive substances decreased in melanoma samples in direct relation to the Clark levels. Higher glutathione S-transferase activity, particularly in the most invasive forms of melanoma, indicates that this type of cancer is more malignant. Similarly, a decrease in superoxide dismutase activity can also encourage progression of the tumor. These results are in accord with those from tumor cell cultures and could suggest new strategies (gene therapy) for managing skin cancer.

Parameters related to oxygen free radicals in erythrocytes, plasma and epidermis of the hairless rat.

The following parameters related to oxygen free radicals (OFR) were determined in erythrocytes and the epidermis of hairless rats: catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), reduced (GSH) and oxidized (GSSG) glutathione, glutathione S-transferase (GST), superoxide dismutase (SOD) and thiobarbituric acid reactive substances (TBARS). GSH, GSSG and TBARS were also analyzed in plasma. In erythrocytes, the Pearson correlation coefficients (r) were significant (p < 0.001) between glutathione and other parameters as follows: GSH correlated negatively with GSSG (r = -0.665) and TBARS (r = -0.669); GSSG correlated positively with SOD (r = 0.709) and TBARS (r = 0.752). Plasma GSSG correlated negatively with erythrocytic thermostable GST activity (r = -0.608; p=0.001) and with erythrocytic total GST activity (r = -0.677; p < 0.001). In epidermis (p < 0.001 in all cases), GSH content correlated with GSSG (r = 0.682) and with GPx (r = 0.663); GSSG correlated with GPx (r = 0.731) and with GR (r = 0.794). By multiple linear regression analysis some predictor variables (R(2)) were found: in erythrocytes, thermostable GST was predicted by total GST activity and GSSG, GSSG content was predicted by GSH and by the GSH/GSSG ratio and GPx activity was predicted by GST, CAT and SOD activities; in epidermis, GSSG was predicted by GR and SOD activities and GR was predicted by GSSG, TBARS and GPx. It is concluded that the hairless rat is a good model for studying OFR-related parameters simultaneously in blood and skin, and that it may provide valuable information about other animals under oxidative stress.

Glutathione, glutathione S-transferase and reactive oxygen species of human scalp sebaceous glands in male pattern baldness.

We investigated the contribution of reactive oxygen species to the development of sebaceous gland hyperplasia and the characteristics of the glutathione S-transferase/glutathione system in male pattern baldness. Glutathione S-transferase, glutathione, and thiobarbituric acid-reactive substances were determined in sebaceous gland-enriched scalp skin of men affected by male pattern baldness and were subjected to hair autotransplantation. In comparison with the hairy occipital-donor areas, the following results were obtained in alopecic frontoparietal samples: glutathione S-transferase-specific activity increased 7-fold (p < 0.001); enzyme affinity towards 1-chloro-2,4-dinitrobenzene decreased 2-fold (p = 0.009); glutathione content decreased 2.5-fold (p = 0.017); and thiobarbituric acid reactive substances increased 2-fold (p = 0.006). Chromatofocusing analysis, bromosulfophthalein IC50 values, enzyme-linked immunosorbent assay, and immunohistochemistry with polyclonal antibodies raised against glutathione S-transferases alpha, mu, and pi demonstrated the presence of alpha, pi, and probably the 5.8 alpha isoenzymes in the sebaceous gland. These results support the hypothesis that reactive oxygen species are involved in the pathogenesis of sebaceous gland hyperplasia in male pattern baldness.

[The kinetic characteristics of the erythrocyte glutathione S-transferase system as a function of sex and the tobacco habit]. / Características cinéticas del sistema glutatión S-transferasa eritrocitario en función del sexo y del hábito tabáquico.

BACKGROUND: Erythrocytic glutathion S-transferase (GST) plays an important role as a protective mechanism against oxidative stress. The present study was conducted to evaluate the influence of both smoking habit and sex upon the kinetic characteristics of the enzyme. SUBJECTS AND METHODS: 176 healthy subjects (100 men and 76 women), smokers and nonsmokers, were included. Enzyme parameters were calculated in erythrocytic haemolysates using 1-chloro-2,4-dinitrobenzene (CDNB) and glutathion (GSH) as substrates. Haemoglobin (Hb) was removed by affinity chromatography. In samples coming from 51 men and 42 women the native haemolysate was subjected to thermal shock (52 degrees C) and the enzyme parameters were compared with those obtained in the non-denatured samples. RESULTS: In non-denatured samples, Km (mM) and Vmax (mumol/min/g Hb) values for CDNB were significantly higher (p < 0.001) in smokers than in non smokers, especially in women. Thus, respectively for Km and Vmax (mean +/- standard deviation): for men non smokers, 1.43 +/- 0.54, 1.63 +/- 0.42 and smokers, 1.74 +/- 0.5, 1.8 +/- 0.69; for women, non smokers 1.42 +/- 0.56, 1.57 +/- 0.46 and smokers, 2.05 +/- 0.59, 2.51 +/- 0.6. Thermal denaturation diminished the enzyme activity in all cases and modified the Km values, these results were opposite to those obtained in the non-denatured samples. Thus, for Km and Vmax respectively: for men, smokers, 1.6 +/- 0.71 and 0.9 +/- 0.32 and non smokers, 1.4 +/- 0.66 and 0.53 +/- 0.29; for women, non smokers, 2.00 +/- 0.58, 1.13 +/- 0.29 and smokers 1.22 +/- 0.77, 0.52 +/- 0.23. The GSt content was similar in the four groups studied (3.75 +/- 1.15 mumol SH/g Hb). CONCLUSIONS: The greater thermolability of GST activity and the increase in the Km values observed in smokers, especially in women, should be considered as indicative of an increased risk for the erythrocytes against oxidative stress.

Glutathione S-transferase in normal human anagen hair follicles.

Glutathione S-transferase (GST) has been quantified and characterized in healthy human anagen hair follicles obtained from 36 men and 36 women (26 +/- 7 years of age). GST activity was determined using 1-chloro-2,4-dinitrobenzene as a substrate, and the values in men were: 0.5 +/- 0.2 mU/follicle, significantly different from women (0.36 +/- 0.2 mU/follicle); 196 +/- 98 mU/mg protein and 309 +/- 158 mU/mg DNA without significant differences from women. Enzyme activity showed a high degree of inter-individual variability (23.5-fold when expressed per follicle, 18.29-fold expressed per mg of protein and 22.75-fold per mg of DNA) in the whole population and this variability was higher in women. Ion-exchange chromatography by KCl and enzyme immunoassay suggest that the GST present in hair follicles corresponds with the acidic form. The percentage of anagen hairs in each subject showed a positive correlation with the following parameters: GST/hair, GST/DNA and DNA/hair. It is concluded that GST may contribute to the maintenance of the hair growth cycle.

Modifications of glutathione S-transferase (GST) activity in the last period of pregnancy in rats treated with benzo(a)pyrene (BP).

Pregnant rats were treated with benzo(a)pyrene (BP) (50 mg/kg every 2 days) from day 7 of pregnancy and killed at day 16 or day 19. Km of erythrocyte glutathione S-transferase (GST) decreased during pregnancy in control rats (1.29 x 10(-3) M at day 16; 1.02 x 10(-3) M at day 19) and even more in treated rats at day 19 (0.71 x 10(-3) M). Vmax was lower in treated rats at day 19 (0.56 mumol/min/g haemoglobin) than in control rats (0.88 mumol/min/g haemoglobin) suggesting inhibition of the enzyme. Placental weight diminished in treated rats at day 19 but was not affected at day 16. Chromatofocusing of placental GST showed a single peak (pH 8.3-8.6) in control and treated rats on day 16 and an additional peak (pH 7.0-7.4) in treated rats on day 19. An increase in Km (2.84 x 10(-3) M) and Vmax (69 nmol/min/mg protein) in placental GST was observed in treated rats at day 16 (Km = 1.61 x 10(-3) M; Vmax = 43.3 nmol/min/mg protein, in control rats) followed by a decrease in these parameters in rats treated until day 19 (Km = 1.63 x 10(-3) M; Vmax = 48.7 nmol/min/mg protein). These results suggest that BP, initially, stimulates GST synthesis in placenta, followed by an inhibition of the enzyme at day 19. Fetal weight was also affected by BP treatment, especially at day 16. Km and Vmax values of fetal GST were higher in treated rats at day 16 than in control rats but these differences were not detectable at day 19. This may be explained by the more protective role of the placenta at day 19 than at day 16. Glutathione content in erythrocytes, placenta and fetus was not affected by BP.

Air pollution and urinary thioether excretion in children of Barcelona.

The polluted environment found in highly industrialized areas and in big cities contains a great quantity of electrophilic (EC) and proelectrophilic (PEC) compounds, which largely contribute to the development of several pathological processes in humans. EC and PEC can be coupled to the cysteine moiety of glutathione spontaneously or by the glutathione S-transferase system (GST), giving nontoxic metabolites that can be eliminated as urinary thioethers (UT). Therefore one approach to establishing the degree of impact of EC and PEC on the human body is the analysis of UT in the population living in polluted environments. The work presented here has been carried out in the city of Barcelona with a group of 50 children living in a polluted area, over a 12-mo period. Our results demonstrate that UT are significantly higher when the amounts of air pollutants (AP) increase; although the level of contamination never exceeded the established "safe limits," UT reached values similar to those found in people smoking more than 10 cigarettes/d. These results may contribute to establishing the maximal levels of contamination compatible with a healthy life, on the basis of patterns of true salubrity rather than on political and economic criteria.