Nilson Penha-Silva

Influence of age on the stability of human erythrocyte membranes

This work evaluated the dependence of erythrocyte membrane stability on age, temperature (26, 32, 37, 42 and 47 degrees C), nutritional status, red cell count, red cell distribution width (RDW) and mean corpuscular volume (MCV) in human females (n = 67, 20-94 years). Erythrocyte membrane resistance to hypotonic lysis was expressed as the NaCl concentration (H(50)) capable of promoting 50% haemolysis. A nutritional evaluation was performed using the Mini Nutritional Assessment (MNA) instrument, which showed that 76.1% of the study group were well nourished and 23.9% were at risk of malnutrition. H(50) demonstrated a negative correlation with age at all temperatures. H(50) showed no correlation with either MNA scores or with any haematological indices. The power and significance of the correlations improved when we censored from the analyses those individuals at risk of malnutrition. The thermal dependencies lines for H(50) demonstrated higher values for females 20-39 years of age compared with those more than 60 years of age. These results suggest that erythrocyte resistance to hypotonicity (erythrocyte membrane stability), increased with age of study volunteers. This increased stability was more pronounced among well-nourished individuals.

Purification and biochemical characterization of Eumiliin from Euphorbia milii var. hislopii latex.

A protease, which we designate Eumiliin, was isolated from the latex of Euphorbia milii var. hislopii by a combination of ion-exchange chromatographic steps using DEAE-Sephacel and gel-filtration with Sephadex G-75. Eumiliin is a monomeric protein with an apparent molecular mass of 30 kDa by SDS-PAGE under reducing conditions and gave one main peak at 29,814 KDa in MALDI-TOF/TOF mass spectrometry. Eumiliin has caseinolytic and fibrinogenolytic activities, but no hemorrhagic or defibrinating activities. The enzyme readily hydrolyzes the Aalpha-chain of fibrinogen and, more slowly, the Bbeta-chain. Its fibrinogenolytic activity is inhibited by beta-mercaptoethanol and leupeptin. In contrast, EDTA and benzamidine did not affect the activity of Eumiliin. The caseinolytic activity of Eumiliin had a pH optimum of 8.0 and was stable in solution at up to 40 degrees C; activity was completely lost at >or=80 degrees C. Intraplantar injection of Eumiliin (1-25 microg/paw) caused a dose- and time-dependent hyperalgesia, which peaked 1-5h after enzyme injection. Intraplantar injection of Eumiliin (1-25 microg/paw) also caused an oedematogenic response that was maximal after 1h. Morphological analyses indicated that Eumiliin induced an intense myonecrosis, with visible leukocyte infiltrate and damaged muscle cells 24h after injection.

Bhalternin: Functional and structural characterization of a new thrombin-like enzyme from Bothrops alternatus snake venom

A serine protease from Bothrops alternatus snake venom was isolated using DEAE-Sephacel, Sephadex G-75 and Benzamidine-Sepharose column chromatography. The purified enzyme, named Bhalternin, ran as a single protein band on analytical polyacrylamide gel electrophoresis (SDS-PAGE) and showed molecular weights of 31,500 and 27,000 under reducing and non-reducing conditions, respectively. Its complete cDNA was obtained by RT-PCR and the 708bp codified for a mature protein of 236 amino acid residues. The multiple alignment of its deduced amino acid sequence showed a structural similarly with other serine proteases from snake venoms. Bhalternin was proteolytically active against bovine fibrinogen and albumin as substrates. When Bhalternin and bovine fibrinogen were incubated at 37 degrees C, at a ratio of 1:100 (w/w), the enzyme cleaved preferentially the Aalpha-chain, apparently not degrading the Bbeta and gamma-chains. Stability tests showed that the intervals of optimum temperature and pH for the fibrinogenolytic activity were 30-40 degrees C and 7.0-8.0, respectively. Also, the inhibitory effects of benzamidine on the fibrinogenolytic activity of Bhalternin indicate that it is a serine protease. This enzyme caused morphological alterations in heart, liver, lung and muscle of mice and it was found to cause blood clotting in vitro and defibrinogenation when intraperitoneally administered to mice, suggesting it to be a thrombin-like enzyme. Therefore, Bhaltenin may be of interest as a therapeutic agent in the treatment and prevention of thrombotic disorders.

Dependence of Mini-Nutritional Assessment Scores with Age and Some Hematological Variables in Elderly Institutionalized Patients

Background: Nutritional imbalance is a serious problem in developing countries, especially for the older population, which makes simple and rapid instruments for nutritional evaluation very necessary in order to have a detailed picture of the undernutrition epidemiology in those places. Objective: This work aimed to evaluate the nutritional status of elderly institutionalized patients by use of the Mini-Nutritional Assessment (MNA) test and compare the MNA scores with some hematological variables. Methods: We performed a cross-sectional study in elderly institutionalized patients (153) of all nursing homes in the Brazilian city of Uberlândia, using the MNA questionnaire and quantitative analysis of erythrocytes, hematocrit, hemoglobin, serum iron and transferrin. The subjects were classified by gender, nutritional state and age range. The nutritional categories were compared by analysis of variance and the dependence between the considered variables was tested with the Spearman correlation analysis. Results: 60% of the subjects were female. The average age was 74.6 ± 9.5 and 78.5 ± 9.5 years for male and female individuals, respectively. 18.3% of the patients presented undernutrition, 45.7% presented risk of undernutrition, and 36.0% were well nourished. These three nutritional categories showed statistically significant differences among the MNA scores, age and all the hematological parameters, for the sum of both genders, but not for the age, iron and transferrin values of the male individuals and hematocrit and hemoglobin values of the female patients. Significant differences between genders were not observed only for the MNA and transferrin values. MNA values presented positive correlations with erythrocytes, hematocrit, hemoglobin, and serum iron levels, and also a negative correlation with age, which was attributed to a worsening of the nutritional state with age. Conclusion: Based on the MNA, 64% of the elderly institutionalized patients of Uberlândia presented risk of undernutrition and undernutrition, and their nutritional state is aggravating with the age increase, which is consistent with the age-dependent decrease observed in some hematological variables. Since the low MNA scores were not necessarily associated to subnormal mean values of the hematological variables, we can conclude that the low MNA values are predicting situations that still did not manifest pathologically in those variables.

Effects of glycerol and sorbitol on the thermal dependence of the lysis of human erythrocytes by ethanol.

In this work, the effects of 1 mol/L glycerol or sorbitol on the thermal dependence (27-47 degrees C) of the lysis of human erythrocytes by ethanol in saline solution (0.154 mol/L NaCl) have been evaluated. Lysis was monitored by measurement of the absorbance at 540 nm. Ethanol produced either lysis or protection against lysis depending on the conditions. These antagonistic effects are attributed to the existence of expanded (R) and compacted (T) erythrocytes, present under conditions of low and high osmolarity, respectively. The transitions of lysis of the R state and formation and lysis of the T state were all found to be sigmoidally defined. The ethanol concentration at the midpoint of the lysis transition of the R state (D(50R)) was found to decrease with increasing temperature and osmolarity. In the presence of glycerol or sorbitol, an increase in temperature led to smaller decreases in D(50R) and osmotic protection against lysis. The ethanol concentration at the midpoint of formation (S(50T)) and lysis (D(50T)) of the T state also decreased with increasing temperature and osmolarity. Lysis of R state erythrocytes is determined by the chaotropic action of ethanol, but the formation and lysis of T state erythrocytes are determined by osmotic pressure effects.

Efeito da composição do solvente sobre a estabilidade de proteínas em soluções aquosas

A protein presents a native (N) macro state, which is functionally active, in equilibrium with the denatured (D) macro state, which is devoid of biological activity. An ensemble of microstates forms each macrostate. The denatured state comprises a greater ensemble of microstates than the native macrostate. The N-D equilibrium can be affected by several factors, that comprise the purity of the water, temperature, pH and solute concentration. This work discusses the influence of osmolytes and chaotropics on the N-D equilibrium in aqueous solutions.

Abdominal Obesity and Association With Atherosclerosis Risk Factors: The Uberlândia Heart Study

AbstractEctopic visceral fat (VF) and subcutaneous fat (SCF) are associated with cardiovascular risk factors. Gender differences in the correlations of cardiovascular disease risk factors and ectopic fat in the Brazilian population still lacking.Cross-sectional study with 101 volunteers (50.49% men; mean age 56.5 ± 18, range 19–74 years) drawn from the Uberlândia Heart Study underwent ultrasonography assessment of abdominal visceral adipose tissue with convex transducer of 3.5 MHz of frequency. The thickness of VF was ultrasonographically measured by the distance between the inner face of the abdominal muscle and the posterior face of abdominal aorta, 1 cm above the umbilicus. The SCF thickness was measured with a 7.5 MHz linear transducer transversely positioned 1 cm above the umbilical scar. The exams were always performed by the same examiner. Ectopic fat volumes were examined in relation to waist circumference, blood pressure, and metabolic risk factors.The VF was significantly associated with the levels of triglycerides (P < 0.01, r = 0.10), HDL cholesterol (P < 0.005, r = 0.15), total cholesterol (P < 0.01, r = 0.10), waist circumference (P < 0.0001, r = 0.43), systolic blood pressure (P < 0.001, r = 0.41), and diastolic blood pressure (P < 0.001, r = 0.32) in women, and with the levels of triglycerides (P < 0.002, r = 0,14), HDL cholesterol (P < 0.032, r = 0.07), glucose (P < 0.001, r = 0.15), alanine aminotransferase (ALT) (P < 0.008, r = 0.12), gamma-GT (P < 0.001, r = 0.30), waist circumference (P < 0.001, r = 0.52), systolic blood pressure (P < 0.001, r = 0.32), and diastolic blood pressure (P < 0.001, r = 0.26) in men. SCF was significantly associated with the levels of triglycerides (P < 0.01, r = 0.34), LDL cholesterol (P < 0.001, r = 0.36), total cholesterol (P < 0.05, r = 0.36), waist circumference (P < 0.0001, r = 0.62), systolic and diastolic blood pressure (P < 0.05, r = 0.34) in women, and with the waist circumference (P < 0.001, r = 0.065)), and MetS (P < 0.05, r = 0.11) in men.The VF and SCF were correlated with most cardiovascular risk factors in both genders but our findings support the idea that there are gender differences in the correlations between ectopic fat deposition and the cardiovascular risk factors.

Influence of age on the correlations of hematological and biochemical variables with the stability of erythrocyte membrane in relation to sodium dodecyl sulfate

Abstract Objectives To evaluate the influence of age on the relationships between biochemical and hematological variables and stability of erythrocyte membrane in relation to the sodium dodecyl sulfate (SDS) in population of 105 female volunteers between 20 and 90 years. Methods The stability of RBC membrane was determined by non-linear regression of the dependency of the absorbance of hemoglobin released as a function of SDS concentration, represented by the half-transition point of the curve (D50) and the variation in the concentration of the detergent to promote lysis (dD). Results There was an age-dependent increase in the membrane stability in relation to SDS. Analyses by multiple linear regression showed that this stability increase is significantly related to the hematological variable red cell distribution width (RDW) and the biochemical variables blood albumin and cholesterol. Discussion The positive association between erythrocyte stability and RDW may reflect one possible mechanism involved in the clinical meaning of this hematological index.

Evolution of nutritional, hematologic and biochemical changes in obese women during 8 weeks after Roux-en-Y gastric bypasss

Obesity is a chronic disease of multifactorial origin and currently is a serious public health problem. The treatment of morbid obesity can be effectively done by bariatric surgery. The present study aimed to evaluate the influence of changes in food intake on body composition and some hematologic and biochemical variables in the period of eight weeks after Roux-en-Y gastric bypass (RYGB). The study included 22 women submitted to RYGB. We evaluated anthropometric, nutritional, hematologic and biochemical variables before and 14, 28, 42 and 56 days after surgery. The patients showed a decrease in caloric intake and hence macro- and micronutrients, with significant loss of weight and decrease in body mass index (BMI). Decreases in body weight and BMI were associated with reduced blood levels of total cholesterol, VLDL-C, LDL-C, triglycerides and glucose with time after surgery. The decrease in caloric intake was also associated with decreased intake of protein, iron and calcium, with a decline in hemoglobin, hematocrit and red blood count, and RDW increased after surgery.

Bivariate and multivariate analyses of the correlations between stability of the erythrocyte membrane, serum lipids and hematological variables

The observation that the fluidity must remain within a critical interval, outside which the stability and functionality of the cell tends to decrease, shows that stability, fluidity and function are related and that the measure of erythrocyte stability allows inferences about the fluidity or functionality of these cells. This study determined the biochemical and hematological variables that are directly or indirectly related to erythrocyte stability in a population of 71 volunteers. Data were evaluated by bivariate and multivariate analysis. The erythrocyte stability showed a greater association with hematological variables than the biochemical variables. The RDW stands out for its strong correlation with the stability of erythrocyte membrane, without being heavily influenced by other factors. Regarding the biochemical variables, the erythrocyte stability was more sensitive to LDL-C. Erythrocyte stability was significantly associated with RDW and LDL-C. Thus, the level of LDL-C is a consistent link between stability and functionality, suggesting that a measure of stability could be more one indirect parameter for assessing the risk of degenerative processes associated with high levels of LDL-C.