Extração, caracterização e atividades biológicas de proteínas da espécie cnidoscolus urens (L.) Arthur

The extraction, chemical and structural characterization of a wide variety of compounds derived from plants has been a major source of bioactive molecules. Several proteases have been isolated in the plant kingdom, with numerous pharmacological and biotechnological applications. Among the proteases isolated from plants, are the fibrinogenolytic, with relevant application in the treatment of disorders in the coagulation cascade, in addition to potential use as a tool in clinical laboratories. In this study, in addition to evaluating the effects of the protein extract of Cnidoscolus urens (L.) Arthur (Euphorbiaceae) in the coagulation cascade also investigates the presence of antimicrobial activity and characterizes the proteolytic activity detected in this extract, aiming to determine their potential pharmacological and biotechnological application. In this way, crude protein extracts obtained from the leaves of C. urens in Tris-HCl 0.05M, NaCl 0.15M, pH 7.5, were precipitated in different concentrations of acetone, and assessed for the presence of proteolytic activity in azocaseína and fibrinogen. The most active fraction (F1.0) in these tests was chosen for assessment of biological activity and biochemical characterization. The Aα chain and Bβ of fibrinogen were completely cleaved at a concentration of 0.18 μg/μL of protein fraction in 4 minutes. Fibrinogenolytic activity presented total inhibition in the presence of E-64 and partial in the presence of EDTA. The fraction demonstrated coagulant activity in plasm and reduced the APTT, demonstrating acting on the factors coagulation of the intrinsic pathway and common, not exerting effects on the PT. Fibrinolytic activity on plasma clot was detected only in SDS-PAGE in high concentrations of fraction, and there were no defibrinating. Although several proteases isolated from plants and venomous animals are classically toxic, the fraction F1.0 of C. urens not expressed hemorrhagic nor hemolytic activities. Fraction F1.0 also showed no antimicrobial activity. In proteolytic activity on the azocasein, the optimal pH was 5.0 and optimum temperature of 60ºC. The enzyme activity has been shown to be sensitive to the presence of salts tested, with inhibition for all compounds. The surfactant triton did not influence the enzyme activity, but the tween-20 and SDS inhibited the activity. In the presence of reducing agents increase in enzyme activity occurred, a typical feature of enzymes belonging to the class of cysteine proteases. Several bands with proteolytic activity were detected in zymogram, in the region of high-molecular-weight, which were inhibited by E-64. In this study, we found that C. urens presents in its constitution cysteine proteases with fibrinogenolytic and procoagulant activity, which may be isolated, with potential application in treatment of bleeding disorders, thrombolytic and clinical laboratory

Efetividade da terapia fisica descongestiva na cicatrização de úlceras venosas

A venous ulcer is the most serious clinical manifestation of chronic venous failure, and the most frequent in lower limbs, accounting for 70% of all ulcers. Patients may suffer from this infirmity for several years without healing of the lesion if treatment is inadequate. The aim of this investigation was to verify the effectiveness of decongestive physical therapy in the healing of venous ulcers. This is a quasi-experimental, interventionist study, with paired, non-probabilistic sampling, composed of 50 patients divided into two groups: control and intervention, each composed of 25 patients. Both groups were identically treated for six months with daily dressings and the latter also underwent complex physical therapy consisting of a combination of the following techniques: manual lymphatic drainage, compression bandaging, lower limb elevation, myolymphokinetic exercises and skin care. The study was approved (Protocol no. 59/2007) by the Ethics Committee of the State University of Southeast Bahia. The Mann-Whitney and Chi-square tests were applied for data analysis. After statistical analysis the patients who underwent therapy showed a statistically significant difference with respect to wound contraction starting in the second month of treatment. Compared to the control, the intervention group showed a greater reduction in both pain and edema starting in the third and fourth month of therapy, respectively. To reinforce these findings, the mean percentage of tissue present at the base of the ulcer (granulation/fibrin ratio) was calculated. The intervention group showed greater granulation at the base of the ulcer compared to the control, significant from the second month of treatment on. It was observed in this study that lymphotherapy, when compared between the intervention and control groups, accelerated the healing process, and reduced pain and edema in the affected limbs. It is expected, therefore, that these results widen scientific knowledge and we suggest that this therapy be used 78 not only to reduce lymphedema, but also as a treatment option for venous ulcers, given its easy application and low cost. The data, therefore, demonstrated the importance of basic care on the part of a multiprofessional and transdisciplinary health team involved in the healing process of these infirmities, thereby contributing to better quality of life in these individuals

Qualidade de vida de pessoas com úlcera venosa: associação dos aspectos sociodemográficos, de saúde, assistência e clínicos da lesão

Venous ulcer (VU) is a lower limbs injury resulting from inadequate return of venous blood in feet or legs. Although it is not a deadly disease, it causes chronic wounds, which seriously undermine patients´ quality of life (QOL) and sometimes leads to drastic family, social, economic and psychological changes. In this sense, there are several aspects that may influence the venous ulcers patients´ QOL. The study´s objective aimed on the association of socio-demographic and health, health care and clinical injury on UV patients‟ QOL. Analytical studies, which consider the complexity of factors involved in changes in UV patients‟ QOL has a cross-sectional and quantitative approach. The HUOL Ethics Committee approved this project (n.279/09). The collection of data lasted a period of 3 months in 2010 and it took place at the clinic of Angiology at Hospital Universitário Onofre Lopes (HUOL). The data sample consisted of 60 patients treated by UV angiologists in the HUOL Surgical Clinic. The results were analyzed with SPSS 15.0 by descriptive and inferential statistics. The study was based on UV patients that were predominantly female, average age of 61.4 years, that had low education level and low family income, with occupations requiring long periods of standing or sitting, but mostly retired, unemployed or laid off due to the disease and/or due to chronic diseases associated with the UV. The study took also into consideration patients that used inappropriate products, that were improperly treated by a professional caregiver, that lacked of adequate guidance and compression therapy, that performed no lifting of the lower limbs and regular exercise, that the time of injury were greater than or equal to six months, that were missing specific laboratory tests. The study‟s reference were on recurrent lesions, medium to large lesions area, bed of the lesion (injuries) with fibrin and/or necrosis, with amount of exudate with medium to large, odorless and no signs of infection, with tissue loss between 1st and 2nd degree, without collecting swab or biopsy and with pain. In general, QOL of researched individuals were considered low, the maximum score was 69 points, which the areas that were mostly influenced were the total scores of QOL functional capacity (0.021), emotional (0.000) and social functioning (0.080). Of the 60 individuals, 53.3% had scores between 40 and 69 points in SF-36, and they had the best scores in sociodemographic and health variables (ρ = 0.049). In respect to the assistance and injury characteristics, patients who scored between 40 and 69 points in SF-36 had better scores on these characteristics. By combining the socio-demographic variables, health, and handling characteristics of the injury, we observed a significant difference (ρ = 0.032) when linking them with the QOL total scores. When analyzing separately the domains of the SF-36 scores on the quality of life, we find that the areas that showed statistical significance were functional ability (ρ = 0.035), appearance (ρ = 0.019), emotional (ρ = 0.000), and mental health (ρ = 0.050). Among the socio-demographic characteristics studied, gender and marital status contributed more to the reduction of QOL and among the variables of assistance and the injury, orientation, reference and area of UV contributed the most. By analyzing these five variables all together in accordance with the overall score obtained in the quality of life, we found a significant correlation (ρ = 0.002); with 6.23 times more chances of patients have better QOL in the presence of these five positive factors. By conducting the Mann Whitney U test between all the five demographic variables, health, and clinical care, we found that this combination also proved to be significant (ρ = 0.006). Therefore, patients with these five variables positive tend to have a better QOL. Based on these results, we reject the null hypothesis (H0) and accept the alternative hypothesis (H1) proposed in this study because we noted that the QOL of patients with UV is associated with sociodemographic and health, health care and clinical aspects of the injury