Biomedical subjects

A Smolen

Publications and source records attributed to A Smolen.

45 records · Page 3Linked to original sources

PubMedDate not supplied

Pyridoxal 5'-phosphate and pyridoxamine 5'-phosphate concentrations in blood and tissues of mice fed ethanol-containing liquid diets.

The effects of chronic ethanol administration on vitamin B-6 metabolism were studied in female Long-Sleep (LS) and Short-Sleep (SS) mice. Animals were fed an ethanol containing liquid diet (AIN-76) for four weeks. Concentration of ethanol in the diet increased from 10 to 25% ethanol-derived calories (EDC) during weeks 1-3 and was maintained at 30% EDC for 1 additional week. We measured concentrations of pyridoxal 5'-phosphate (PLP) in plasma, erythrocytes and whole blood, and liver and brain PLP and pyridoxamine 5'-phosphate (PMP) in ethanol-fed and pair-fed control mice. Chronic ethanol administration significantly increased PMP and total (PLP + PMP) levels in the liver of SS mice. In LS mice ethanol feeding significantly decreased PMP and total (PLP + PMP) levels in the brain, but these values were still within normal limits. These results suggest that both control and ethanol-containing liquid diets are nutritionally adequate with respect to vitamin B-6, and that chronic ethanol administration does not adversely affect vitamin B-6 metabolism in adult mice.

Animals↗

PubMedDate not supplied

Developmental profile of hepatic alcohol and aldehyde dehydrogenase activities in long-sleep and short-sleep mice.

Ethanol is metabolized primarily in the liver by a cytosolic alcohol dehydrogenase (ADH). The product, acetaldehyde, is metabolized to acetate by nonspecific aldehyde dehydrogenases (AHD). Mouse liver contains five major constitutive AHD isoenzymes: mitochondrial high Km (AHD-1), mitochondrial low Km (AHD-5), cytosolic high Km (AHD-7), cytosolic low Km (AHD-2) and microsomal high Km (AHD-3). The Long-Sleep (LS) and Short-Sleep (SS) mice differ in their sleep time response to ethanol as early as 10 days of age, and this difference increases with increasing age. Age- and genotype-related differences in metabolism could account for the pattern of responses seen in these mice. We measured the activity of hepatic ADH and the five AHD isoenzymes in LS and SS mice from 3 days of age to adulthood to determine if there were differences in the developmental profiles of these enzyme activities. We found no sex differences in the developmental profile of either ADH or AHD, and the LS and SS mice have nearly identical ADH and AHD activities with the possible exception of the high Km mitochondrial enzyme activity between days 3 and 6, and the low Km mitochondrial enzyme between days 28 and 32. Thus, it appears that differences in ethanol or acetaldehyde metabolism do not contribute significantly to the differential sensitivity to ethanol between young LS and SS mice or to the differential sensitivity between young and adult mice.

Aging↗

PubMedDate not supplied

Purinergic modulation of ethanol-induced sleep time in long-sleep and short-sleep mice.

The long-sleep (LS) and short-sleep (SS) mice were selectively bred for differences in sensitivity to the depressant effects of ethanol. In addition to their differential sensitivity to ethanol, they are also differentially sensitive to purinergic agonists and antagonists. This suggests that there may be differences in the purinergic systems of these lines of mice which may aid in understanding how they differ in ethanol sensitivity. We have investigated whether these drugs are capable of modifying acute ethanol sensitivity as measured by ethanol-induced loss of the righting response (ethanol sleep time), waking blood and brain ethanol concentrations, and blood ethanol elimination rate. The purinergic agonists cyclohexyladenosine (CHA), L-phenylisopropyladenosine (PIA), 2-chloroadenosine (CAD), and N-ethylcarboxamidoadenosine (NEC) increased sleep time in both LS and SS mice, however, LS mice were generally more affected than SS. The LS and SS mice were also differentially sensitive to the purinergic antagonists, theophylline and caffeine. Blood and brain ethanol concentration on awakening suggested that CNS sensitivity to acute ethanol administration was altered by pretreatment with agonists but not antagonists. Two agonists, CHA and NEC, significantly lowered ethanol elimination in both lines of mice while PIA, CAD, and the antagonists theophylline, and caffeine were without affect on elimination rate. These data support previous observations that adenosine-mediated systems may be involved in the modulation of ethanol sensitivity.

2-Chloroadenosine↗

PubMedDate not supplied

Using logistic regression analysis in preliminary differential diagnosis of adnexal masses.

The aim of our study was to generate predictive model, which would allow to estimate the influence of analyzed parameters on predictive accuracy of differential diagnosis of adnexal masses and to evaluate prospectively diagnostic efficacy of the statistic model in the new set of patients. A total of 686 women diagnosed and surgically treated in the Gynecological and Obstetrical Teaching Hospital of University of Medical Sciences in Poznan, Poland, were recruited into the study. Preoperative diagnostics included gynecological examination, ultrasonographic evaluation, tumor Doppler blood flow analysis, and serum levels of CA125 and TPS. In order to find the best combination of features and to calculate the individual probability of the malignancy, stepwise logistic regression analysis with quasi-Newton estimation was applied. The essential part of the best prognostic model, described by foregoing variables, is as follows: [z = -6.005 + 0.058 x age + 1.174 x septa + 1.317 x tumor localization + 1.185 x ascites + 2.28 x solid element + 2.429 x vessels localization -2.386 x PI + 0.084 x MEDV]. The highest sensitivity and specificity for the obtained model were 87.84% and 93.74%, respectively. Prognostic model, constructed with the use of logistic regression analysis, is characterized by higher sensitivity and specificity than individually applied diagnostic tests. Prospective evaluation of this model application in a larger group of patients with adnexal masses will enable precise assessment of its objective clinical usefulness.

Adnexal Diseases↗

PubMedDate not supplied

Comparison of diagnostic usefulness of predictive models in preliminary differentiation of adnexal masses.

The purpose of this study was to compare prognostic models evaluating the probability of an ovarian cancer occurrence based on a number of clinical and ultrasonographic data in women with adnexal masses. A total of 686 women with adnexal masses underwent the examinations between 1994 and 2002. The recorded parameters included: age, menopausal status, body mass index, the grayscale and Doppler ultrasonographic examination, and selected markers concentration levels. In order to find the best combination of features, which significantly influences the probability of malignancy, stepwise logistic regression analysis, as well as artificial neural network, was used. The diagnostic efficiency of received models was estimated and compared using receiver-operating characteristics (ROC) curve. The results indicate that 431 and 255 patients had a benign and malignant ovarian tumor, respectively. Application of stepwise logistic regression analysis revealed statistically significant importance of eight features. The sensitivity and specificity for the received model were 65.71% and 77.59%, respectively. Three-layer perceptron network shows 13 features as significant predictors of malignancy. The network gave a sensitivity of 85.7% and specificity of 93.1%. Comparison of area under ROC curve for received models was 0.9679 vs 0.9716. Prognostic values of the analyzed neural model are not optimal but seem to surpass logistic regression model in terms of the predictive possibilities.

Adult↗

PubMedDate not supplied

Brain NaK-ATPases in mice differentially sensitive to alcohols.

The isozymes of NaK-ATPase were studied in the particulate fraction of homogenates prepared from cortex of LS and SS mice, two lines of mice that have been selectively bred for differential response to ethanol. In addition to a ouabain-insensitive Mg-ATPase, ouabain dose-response curves have suggested the presence, in brain, of two NaK-ATPase activities with different sensitivities to ouabain. These are designated low Ki (4 X 10(-7) M) and high Ki (2 X 10(-4) M). Ethanol differentially inhibited the ATPases: The ouabain-insensitive activity was less sensitive to ethanol inhibition than were the two ouabain-inhibitable activities. The low Ki (brain specific) NaK-ATPase was more sensitive than was the high Ki activity. However, ethanol inhibited all three components of the ATPase activity in an identical fashion in the two mouse lines. The low Ki activity was also more labile to thermal and p-hydroxymercurobenzoate denaturation than was the high Ki activity. These measures did not differ between the LS and SS lines. SDS-polyacrylamide electrophoresis of particulate fraction of cortical homogenates obtained from LS and SS mice revealed the presence of two protein bands with similar 32P labeling in both lines. Given that electrophoretic pattern and heat or p-hydroxymercurobenzoate inhibition were identical, it seems unlikely that differences in ATPase activity, or inhibition by ethanol, are responsible for the different response of LS and SS mice to ethanol.

Animals↗

PubMedDate not supplied

Dose and route of administration alter the relative elimination of ethanol by long-sleep and short-sleep mice.

The elimination of ethanol following the intravenous and intraperitoneal administration of five ethanol doses was measured in two mouse lines, the long-sleep (LS) and short-sleep (SS), that differ markedly in central nervous system sensitivity to ethanol. Both dose and route of administration affected the relative ethanol elimination in these two mouse lines. When the ethanol was given i.v., a steady increase in elimination rate was seen in the SS mice as the dose was increased. In the LS mice, consistent changes in rate with dose were not seen. At the lower doses tested, the LS eliminated ethanol more rapidly than did the SS but at higher doses the mouse lines did not differ. When the ethanol was given i.p., increases in rate with dose were not seen. Rather, the SS elimination rate remained constant as the dose was increased, whereas in the LS a decrease in rate was observed. As a consequence, the SS eliminated ethanol more rapidly than did the LS at higher doses. A potential explanation for these results is that ethanol elimination, following the administration of large doses, is affected by both enzymological factors and central nervous system sensitivity to ethanol. The central nervous system factors may be mediated via ethanol-induced hypothermia.

Alcohol Oxidoreductases↗

PubMedDate not supplied

Purification of human serum paraoxonase/arylesterase. Evidence for one esterase catalyzing both activities.

Evidence is presented that human serum contains a single enzyme with both paraoxonase and arylesterase activities. Throughout the steps of purification and after obtaining over 600-fold purification of the enzyme, the arylesterase activity (measured with phenylacetate as the substrate) co-eluted and retained the same ratio of activity to paraoxonase activity as it had in the initial plasma sample. Paraoxon and DFP (diisopropylfluorophosphate) both complete with phenylacetate as substrates; the inhibition is of mixed type with paraoxon and competitive with DFP. Paraoxonase and arylesterase activities require calcium, and both are inhibited to the same degree by EDTA. Purified arylesterase/paraoxonase is a glycoprotein with a minimal molecular weight of about 43,000. It has up to three sugar chains per molecule, and carbohydrate represents about 15.8% of the total weight. The enzyme has an isoelectric point of 5.1. Its amino acid composition shows nothing unusual, except for a relatively high content of leucine. We conclude that human serum arylesterase and paraoxonase activities are catalyzed by a single enzyme, capable of hydrolyzing a broad spectrum of organophosphate substrates and a number of aromatic carboxylic acid esters. Studies on the genetically determined polymorphism responsible for two allozymic forms (A and B) of the esterase are described in the following paper.

Carboxylic Ester Hydrolases↗

PubMedDate not supplied

Characteristics of the genetically determined allozymic forms of human serum paraoxonase/arylesterase.

Human serum paraoxonase/arylesterase is an esterase with broad substrate specificity. It occurs in two genetically determined allozymic forms, which we have designated types A and B. These allozymes are presumed to be the products of two allelic genes located at the paraoxonase locus on chromosome 7, which is closely linked to the gene for cystic fibrosis. Paraoxonase activity of the B-type isozyme is considerably higher and stimulated more by 1 M NaCl than A-type paraoxonase. The ratio of paraoxonase activity/arylesterase activity of the B-isozyme is about 8, and that of the A-isozyme about 1. Purified isozymes A or B are free of nearly all other serum proteins, and the broad substrate specificity of the serum esterase is preserved after purification. A variety of substrates are hydrolyzed; these include: diisopropylfluorophosphate, soman, sarin, 4-nitro-phenylacetate, 2-nitro-phenylacetate, 2-naphthylacetate, and phenylthioacetate. The isozymic distinctions in kinetic properties and substrate specificity are preserved during purification. It is likely that the allozymes have very similar turnover numbers with phenylacetate (arylesterase activity), but differ considerably in their turnover numbers with paraoxon. Isozymes A and B have about the same minimal molecular weight of 43,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Further detailed studies on the individual isozymic proteins (or the DNA coding for their amino acid sequence) will be required to detect the exact structural differences in the isozymes.

Carboxylic Ester Hydrolases↗