Malondialdehyde production from spermine by homogenates of normal and transformed cells

The oxidation of spermine in vitro by a mixture of polyamine oxidase and diamine oxidase from pig kidney gives rise to malondialdehyde via 3-aminopropanol as the intermediate. Conversely, with spermidine, under similar experimental conditions, no evidence could be obtained for malondialdehyde formation within the limits of sensitivity of the assay (2.0 nmol). The activities of both these enzymes show about a 2-fold increase in normal rat kidney cells (LA31 NRK) transformed by the temperature sensitive mutant of Rous sarcoma virus (LA31) and incubated at the non permissive temperature (39 degrees C) compared to the activities in LA31 NRK at the permissive temperature (33 degrees C). These same enzymatic activities show no temperature dependent changes in normal rat kidney cells (NRK) or in these same cells infected by the wild type virus (NRK B77). In extracts derived from Friend erythroleukemic cells induced to differentiate by dimethyl sulfoxide or hexamethylene bis acetamide, spermine oxidation takes place more efficiently than in non induced cells. A rise in diamine oxidase activity is seen in LA31 NRK (39 degrees C) 12 h after the temperature shift, whereas morphological manifestations of normalcy are seen only at 48 h. The Km of diamine oxidase is 10(-6) M for putrescine and 10(-3) M for 3-aminopropanol. A possible mechanism involving the well documented acetylation of putrescine [23,26] is proposed for diverting intracellular putrescine away from cytosolic diamine oxidase and towards intramitochondrial monoamine oxidase.     

Motilin and human pancreatic polypeptide measured in CSF from alcoholic and non-alcoholic neurological patients

Thirty-one CSF samples from alcoholics and non-alcoholic neurological patients were assayed for immunoreactive motilin and human pancreatic polypeptide (HPP). Both peptides were detected in all samples. Alcoholics without liver disease had significantly higher levels of motilin and lower levels of HPP than neurological controls.     

Active efflux of lithium from erythrocytes of manic-depressive subjects

Lithium transport kinetics, studied under physiological conditions in erythrocytes obtained from manic-depressive patients, are characterized by asymmetric rate constants of efflux and influx. The efflux constants, which are more than twice as large as the influx constants, correlate well with the $\text{in vivo}$ distribution of lithium between erythrocytes and plasma. The efflux process, which is not inhibited by ouabain and is therefore distinct from the sodium-potassium pump, is characterized by Michaelis - Menten kinetics and a large energy of activation. There appears to be a normal endogenous inhibitor which regulates the activity of the postulated lithium pump.     

Platelet serotonin concentration in alcoholic subjects

Serotonin (5-hydroxytryptamine, 5-HT) is assumed to play a role in the pathophysiology of different psychiatric disorders including alcoholism. Since platelets and central serotonergic synaptosomes share similar pharmacodynamics of 5-HT, this study determined platelet 5-HT concentration in 148 male and 42 female drug-free subjects with alcohol dependency, according to the DSM-IV criteria, and in sex-and age-matched controls. Male and female alcoholics had significantly lower platelet 5-HT concentration than 110 male and 123 female healthy controls. Sex differences, i.e. higher platelet 5-HT concentration in men than in women, were found both in healthy and alcoholic subjects. Platelet 5-HT concentration differed significantly in male and female alcoholic subjects with or without different psychiatric comorbidities. Platelet 5-HT concentration was higher in male alcoholics with comorbid posttraumatic stress disorder (PTSD) than in male alcoholics with comorbid anxious-depressive disorder, or depression, or male alcoholics without any psychiatric comorbidities. Comorbid depression in female alcoholics slightly elevated platelet 5-HT levels but these values were still reduced compared to values in healthy women. Smoking status did not affect platelet 5-HT concentration either in healthy or in alcoholic subjects. The data from our study show sex differences, and reduced platelet 5-HT values, regardless of the nicotine dependence, in the large groups of male and female alcoholic subjects. Among male alcoholics the presence of comorbid PTSD partly normalized the decreased platelet 5-HT values. The results of the present study support the hypothesis that alterations in 5-HT system might be related to alcoholism.     

Visual classification of erythrocytes in blood smears from normal subjects and patients

Erythrocytes on specially prepared and coloured blood smears can be classified not only by idcture analysis but also by phase contrast microscopy. Parameters of change for the peripheral zone area and pallor area (phase contrast value PW and index of erythrocyte changes Ev-I) serve as classificators of visual technique. 449 blood smears of the same number of male and female grown-up persons, whose diagnosis was unknown at the time of evaluation, were used for evaluation. In 145 cases these were healthy test persons in clinical and laboratory respect (group I). 163 test persons were ill, but without any malignous neoplasias (group II), and in 141 patients an ensured malignous neoplasia could be found (group III). From those 308 test persons or patients respectively in group I + II 11 blood smears were falsely classified as positive (= 3.6%). In 5 from 141 cases in group III blood smears were falsely evaluated as negative (= 3.6%). The findings were statistically ensured by the t-test. Clinical parameters of evaluation gained with visual techniques of classification are in accordance with those gained with automatic picture analysis. The reasons for the good separating ability of PW and Ev-I are discussed. Visual techniques of classification are suitable for institutions of all grades of equipment.     

Alterations of hemorheological parameters and tubulin content in erythrocytes from diabetic subjects

Treatment of human erythrocytes with high glucose concentrations altered the content and distributions of three tubulin isotypes, with consequent reduction of erythrocyte deformability and osmotic resistance. In erythrocytes from diabetic subjects (D erythrocytes), (i) tubulin in the membrane-associated fraction (Mem-Tub) was increased and tubulin in the sedimentable fraction (Sed-Tub) was decreased, (ii) deformability was lower than in erythrocytes from normal subjects (N erythrocytes), and (iii) detyrosinated/acetylated tubulin content was higher in the Mem-Tub fraction and tyrosinated/acetylated tubulin content was higher in the Sed-Tub fraction, in comparison with N erythrocytes. Similar properties were observed for human N erythrocytes treated with high glucose concentrations, and for erythrocytes from rats with streptozotocin-induced diabetes. In N erythrocytes, high-glucose treatment caused translocation of tubulin from the Sed-Tub to Mem-Tub fraction, thereby reducing deformability and inducing acetylation/tyrosination in the Sed-Tub fraction. The increased tubulin acetylation in these cells resulted from inhibition of deacetylase enzymes. Increased tubulin acetylation and translocation of this acetylated tubulin to the Mem-Tub fraction were both correlated with reduced osmotic resistance. Our findings suggest that (i) high glucose concentrations promote tubulin acetylation and translocation of this tubulin to the membrane, and (ii) this tubulin is involved in regulation of erythrocyte deformability and osmotic fragility.     

Calmodulin and cyclic nucleotide phosphodiesterase activities in erythrocytes from normal and schizophrenic subjects

Calmodulin and cyclic nucleotide phosphodiesterase activities were measured in hemolysates prepared from 18 normal and 17 schizophrenic subjects. No significant difference between groups was found for either activity. The results suggest that calmodulin is present in normal amounts in patients with schizophrenia. This is compatible with the idea that the interaction of calmodulin with antipsychotic agents is structurally non-specific.     

Testosterone production by XYY subjects.

A study of plasma concentration (P1T, ng/ml), metabolic clearance rate (MCRT, L/day) and blood production rate (PBT, mg/day) was done on seven XYY subjects of various ages and four pair-matched control XY subjects by a radioinfusion technique of 1,2-3H-testosterone. Although MCRT showed no significant difference between the groups, P1T and PBT were significantly lower (P less than 0.05) in XYY subjects. Therefore, increased aggressive behavior of the XYY subjects can not be attributed to increased levels or production rates of testosterone.     

Removal of heavy metals from stainless steel effluents by waste biomass from Brazilian alcoholic beverage production

The capacity of waste biomasses from sugar-cane aguardente, a traditional Brazilian spirit, for metal biosorption was assessed. Free biomass and biomass immobilized onto chitin and Dowex (ion-exchange resin) were utilized to remove chromium, iron and nickel from both synthetic solutions and stainless steel effluents. The best performance in terms of metal sorbed was observed in with free biomass, with the following adsorption capacity: 70% chromium, 50% iron and 20% nickel at pH 4.0.     

High-concentration alcoholic production from hydrolysate of raw ground corn by a tetraploid yeast strain

Summary The suitable conditions for high-concentration ethanol production from raw ground corn by a tetraploid yeast strain were examined. We found that the glucoamylase preparation which ia usually employed for alcoholic fermentation of cooked starch could effectively saccharify raw ground corn starch.     

Studies on production of alcoholic beverages from some tropical fruits

The varieties of alcoholic beverages from watermelon; watermelon-banana and watermelon-pineapple mixtures were produced by using monoculture and mixed culture fermentation techniques. Three yeast species, namely, Saccharomyces cerevisiae, Kleochera apicalata, Torulospora delbruckii and four bacterial species Leuconostoc oenos, Lactobacillus Sp, Micrococcus luteus and Streptococcus lactis were identified during the study. The daily succession of these organisms in the various fermenting samples differed in cell mass and occurrence due to their different growth conditions and factors present. A higher bacterial load (3.9 ± 0.2-4.4 ± 0.3) log (cfu) ml^?1 than yeast (2.8 ± 00-4.6 ± 0.4) log (cfu) ml^?1 counts was observed in the mixed culture fermentation, while in the monoculture fermentation, a higher yeast load (4.3 ± 0.3-4.7 ± 0.2) log (cfu) ml^?1 than bacterial loads (2.7 ± 0.1-4.1 ± 0.3) log (cfu) ml^?1 counts was recovered. The results obtained from the present study indicated that monoculture-fermented beverages were of better quality as compared to the mixed culture fermented ones. The monoculture-fermented beverage from watermelon-pineapple mixture was ranked as the best alcoholic beverage based on sensory evaluation score.     

Clinical utility of red cell distribution width in alcoholic and non-alcoholic liver cirrhosis

Red blood cell distribution width (RDW) is a measure of the variation of red blood cell width that is reported as apart of standard complete blood count. Red blood cell distribution width results are often used together with mean corpuscular volume (MCV) results to figure out mixed anemia. The aim of our study was to compare the values of RDW in alcoholic and non-alcoholic liver cirrhosis and to determine if RDW follows the severity of disease according to Child-Pugh score. We retrospectively analyzed 241 patients (176 men and 65 women) with liver cirrhosis and anemia, defined as a hemoglobin value < 130 g/L in men and < 120 g/L in women, which were hospitalized in our Division in a period between 2006 and 2008. Patients were divided in two groups; in first were patients with alcoholic liver cirrhosis, and in second with non-alcoholic cirrhosis. Severity of disease was determined according to Child-Pugh score. Red blood cells distribution width Normal reference range is 11-15%. Alcoholic liver cirrhosis had 204 patients (85%) while non-alcoholic cirrhosis had 37 patients (15%). In group of alcoholic cirrhosis the average RDW was 16.8%. In relation to severity of disease the average RDW for Child-Pugh A was 16.80%, for Child-Pugh B was 16.92%, for Child-Pugh C was 17.10%. In the group of non-alcoholic cirrhosis the average RDW was 16.73% and in relation to severity of disease for Child-Pugh A was 16.25%, for Child-Pugh B 17.01% and for Child-Pugh C was 16.87%. We didn't find statistically significant difference of RDW between alcoholic and non alcoholic cirrhosis (p > 0.05) and we didn't proved any statistically significant increase of RDW in relation to severity of disease in group of alcoholic cirrhosis (p = 0.915) nor in group of patients with non-alcoholic cirrhosis (p = 0.697). Our study showed that RDW had not any clinical value in differentiation of anemia neither in alcoholic and non-alcoholic liver cirrhosis nor in severity of liver disease.     

Increase in glutathione peroxidase activity in erythrocytes from trisomy 21 subjects.

Glutathione peroxidase activity has been measured in erythrocytes from normal subjects and from trisomy 21 patients. The latter cases show about 50 % increase of this enzyme similar to the increase observed for superoxide dismutase (erythrocuprein) suggesting either localisation of the gene for glutathione peroxidase on chromosome 21 (as is the case for erythrocuprein) or regulation of this enzyme by intracellular levels of O₂^$\text{?}$, H₂O₂ or superoxide dismutase.     

The effect of diet on ouabain binding to erythrocytes from obese subjects

Ouabain binding to erythrocyte membranes is increased in obese subjects. Three study groups are compared: 14 reference subjects, 102 +/- 16% of ideal weight; 9 obese on unrestricted diets, 207 +/- 16% of ideal weight; 11 obese on restricted diets, 202 +/- 35% of ideal weight. A reproducible (CV = 11.3%) ouabain-binding assay is used to measure Na+-K+ ATPase sites in erythrocyte membranes. The number of binding sites per red blood cell for obese subjects on unrestricted diets, 431 +/- 30, is greater than for the reference group, 346 +/- 66 (p less than 0.01), or for obese subjects on restricted diets, 371 +/- 68 (p less than 0.05). These data suggest that caloric intake influences the number of Na+-K+ ATPase sites. Scatchard plots indicate only one type of binding site for ouabain with an affinity constant of about 3 X 10(8) M-1.     

Blood thiamine and thiamine phosphate ester concentrations in alcoholic and non-alcoholic liver diseases.

Thiamine state was investigated in patients with alcoholic liver disease, patients with various non-alcoholic liver diseases, and controls using a direct technique (thiochrome assay) to measure thiamine, thiamine monophospate, and the active coenzyme thiamine pyrophosphate in whole blood after isolating the fractions by ion exchange chromatography. Overall nutrition was similar in all groups as assessed by anthropometry, and no patient had clinical evidence of thiamine deficiency. There was no significant difference among the groups in mean concentration of any form of thiamine. The scatter was much greater in patients with alcoholic liver disease but only 8.7% had biochemical thiamine deficiency (defined as a blood concentration of the active coenzyme greater than 2 SD below the mean control value). An unexpected finding was of abnormally high total thiamine concentrations (greater than 2 SD above the mean control value) in 17.4% of patients with alcoholic liver disease, the highest concentrations being found in two patients with severe alcoholic hepatitis and cirrhosis. The ratio of phosphorylated to unphosphorylated thiamine was calculated as an index of phosphorylation and, although the mean did not differ significantly among the groups, the range was greatest in alcoholic liver disease. The lowest ratios occurred in the two patients with severe alcoholic hepatitis, but neither had evidence of thiamine pyrophosphate deficiency. Contrary to studies using indirect assay techniques, these results suggest that thiamine deficiency is unusual in well nourished patients with alcoholic liver disease. The new finding of unexpectedly high thiamine concentrations in some patients may be due to abnormalities of hepatic storage or release in liver disease, particularly in severe alcoholic hepatitis. There was no convincing evidence of impaired thiamine phosphorylation in any patients with liver disease. Conclusions from studies using indirect assays on the prevalence and mechanisms of thiamine deficiency in liver diseases may not be valid.     

Correlation between airway responsiveness and proteoglycan production by bronchial fibroblasts from normal and asthmatic subjects

Asthma is characterized by an airway remodeling process involving altered extracellular matrix deposition such as collagen, fibronectin and proteoglycans. Proteoglycans determine tissue mechanical properties and are involved in many important biological aspects. Not surprisingly, it has been suggested that proteoglycan deposition may alter airway properties in asthma including airway hyperresponsiveness. In chronically inflamed airway tissues, fibroblasts likely represent an activated fibrotic phenotype that contributes to the excessive deposition of different extracellular matrix components. To investigate whether this was the case for proteoglycans, the production of hyaluronan, perlecan, versican, small heparan sulphate proteoglycans (HSPGs), decorin and biglycan was quantified in the culture medium of primary bronchial fibroblast cultures, established from four normal and six asthmatic subjects. Values were further correlated to the airway responsiveness (PC(20) methacholine) of donor subjects. Fibroblasts from subjects with the most hyperresponsive airways produced up to four times more total proteoglycans than cells from subjects with less hyperresponsive or normoresponsive airways. We observed a significant negative correlation between the PC(20) and perlecan, small HSPGs and biglycan, while such correlation was absent for decorin and close to significant for hyaluronan and versican. Altered proteoglycan metabolism by bronchial fibroblasts may contribute to the increased proteoglycan deposition in the bronchial mucosa and to airway hyperresponsiveness characterizing asthma.