The differences between aromatizable and non-aromatizable androgens in relation to body composition and metabolic syndrome risk factors in men.

The relationships between the parameters of metabolic syndrome and non-aromatizable metabolites of testosterone have been discussed in literature. Some papers describe these metabolites as one of the possible causes of male-type obesity. On the contrary, other studies show a protective influence of dihydrotestosterone on visceral obesity. The aim of this study to analyse the relationship between anthropometric parameters, lipid spectrum, glycemia and the level of endogenous testosterone and dihydrotestosterone, and to compare the effects of these androgens. Our population-based study involved 232 healthy men ranging from 20 to 78 years with BMI 18 to 39 kg/m(2). Serum testosterone, dihydrotestosterone and sex hormone binding globulin SHBG levels, lipid spectrum, glucose metabolism parameters were measured and the oral glucose tolerance test was carried out in all subjects. Their anthropometric parameters (weight, height, waist, hips, waist-to-hip ratio, 14 skin folds) and body composition parameters were determined and calculated by the Antropo program. Multiple regression analysis showed a correlation between hormonal levels, esp. of testosterone and dihydrotestosterone, and the anthropometric data, lipid spectrum and parameters of glucose regulation. Low testosterone and/or dihydrotestosterone was correlated to a higher body-mass index, fat content, waist diameter, total-, HDL-, LDL-cholesterol and triglycerides, fasting glucose, insulin resistance and lower muscle and bone mass. In addition, statistical analysis using multivariate regression with reduction in dimensionality did not discover any striking difference between aromatizable and non-aromatizable androgens in their association to lipid and glucose metabolism parameters in healthy, normosthenic men. In conclusion, the association of endogenous testosterone and dihydrotestosterone to anthropometric data, lipid spectrum and insulin sensitivity are of the same quality; however, the effect of the circulating levels of dihydrotestosterone is quantitatively smaller.

Oxidative stress and metabolic syndrome in obese adults with and without controlled diet restriction.

BACKGROUND: Metabolic syndrome (MetS) represents a collection of markers associated with cardiovascular morbidity and mortality. Due to its high prevalence and steady increase of the occurrence, prevention or management of MetS is of paramount importance. The aim of our study was to evaluate MetS occurrence and extent of oxidative stress by comparing obese adults after diet optimization with untreated controls. MATERIAL AND METHODS: Oxidative stress markers (total amount of free radicals, malondialdehyde, allantoin, alpha1-antiproteinase, GSSG/GSH ratio), total antioxidant capacity and lipid standardized alpha-tocopherol were determined in 40 obese people and 48 healthy controls. The obese people were divided into two group A: obese with restricted energy intake with lowered dietary carbohydrates (n=20) and group B: with the same grade of obesity but without following dietary recommendations (n=20). RESULTS: Group A exhibited lower oxidative stress markers than group B; free radicals (5.18+/-1.68 vs 8.43+/-3.66 mmol/l, p<0.01), GSSG/GSH ratio (11.74+/-5.01 vs 15.38+/-5.93%, p<0.05) and higher antioxidants: lipid standardized alpha-tocopherol (3.70+/-0.51 vs 3.35+/-0.60, p<0.05) and ceruloplasmin (0.24+/-0.08 vs 0.21+/-0.03 g/l, p<0.05), in the course of same grade of obesity. Furthermore MetS occurrence was found significantly lower was in group A. CONCLUSION: The energy intake restriction by 2000 kJ, mainly due to carbohydrate limitations, was associated with decreased oxidative stress and simultaneously increased lipid-standardized alpha-tocopherol and ceruloplasmin in obese people. These changes correlated with diminished MetS occurrence by about 50% (Tab. 3, Ref. 32). Full Text (Free, PDF) www.bmj.sk.

The influence of galvanic currents and voltage on the proliferation activity of lymphocytes and expression of cell surface molecules.

Release of metal ions from dental metal fillings supported by galvanism can cause local or general pathological problems in sensitive and genetically susceptible individuals. We aimed to investigate in vitro lymphocyte responses and expression of surface molecules influenced by galvanic currents and voltage. Human peripheral blood lymphocytes were influenced by galvanic currents and voltages and lymphocyte proliferation was measured. Control samples were not exposed to the influence of galvanism. We also studied the expression of surface molecules by the FACS analysis. A 15-h and shorter influence of almost all tested currents and voltages caused a significant decrease in lymphocyte proliferation and the 15-h influence of 20 microA currents significantly increased expression of surface molecules CD 19, 11a/18, 19/69 and 19/95. An influence of 10 and 3 microA currents led to a significant decrease in the expression of surface molecules CD 3, 11a/18, 3/69 and 3/95 and to a significant increase in CD 19 expression. An 80 mV voltage influence led to a significant decrease in the expression of surface molecules CD 3, 11a/18, 3/69, 3/95, 19/69 and 19/95, and 200 and 300 mV voltages significantly decreased the expression of surface molecules CD 3, 19, 11a/18, 3/95 and 19/95 and significantly increased CD 19/69 expression. A long-lasting influence of galvanism can, in sensitive and genetically susceptible individuals, influence lymphocyte proliferation and surface molecule expression. The threshold for pathological values of 5 microA for galvanic currents and 100 mV for galvanic voltage was confirmed.

Antioxidant vitamin levels do not exhibit negative correlation with the extent of acute myocardial infarction.

Serum levels of vitamin E (VE), beta-carotene (BC) and vitamin C (VC) were determined in 50 patients with the first acute myocardial infarction (AMI) before starting thrombolytical treatment. VE and BC were determined by HPLC, VC spectrophotometrically. The reperfused patients were divided according to vitamin concentrations into four groups. The lowest quartile was compared with the rest of the studied population (VE: group with high (H)>15.6 microM>group with low (L), BC: H>0.07 microM>L, VC: H>25 microM>L) in the following parameters: extent of myocardial damage (area under the curves of troponin I, CK-MB during 48 h), arrhythmia and congestive heart failure occurrence, size of ejection fraction, positivity of ventricular late potentials. No significant differences between groups H and L for either VE, BC or VC were found (P 0.05). As no correlation between serum concentrations of vitamins E, C and beta-carotene and the extent and clinical course of AMI was found, the actual vitamin concentrations may be important for prevention of ischemic heart a disease, but they do not play a decisive role in the acute phase of myocardial infarction in humans.

Neuroactive steroids, their precursors and polar conjugates during parturition and postpartum in maternal and umbilical blood: 3.3beta-hydroxy-5-ene steroids.

Five 3beta-hydroxy-5-ene steroids involved in the metabolic route from pregnenolone sulfate to dehydroepiandrosterone and its sulfate, of which three are known allosteric modulators of neurotransmitter receptors, were monitored in the serum of 20 women around parturition. In addition, their levels in maternal and umbilical serum were compared at delivery. On the basis of these data, a scheme of steroid biosynthesis in maternal organism during the critical stages around parturition is proposed. In maternal serum, all the steroids except dehydroepiandrosterone sulfate decreased during labor and even first day after delivery, although their changes were less distinct the more distant from pregnenolone sulfate (PregS) in the metabolic pathway. Calculation of product/immediate precursor ratios in maternal serum over all stages around parturition enabled identification of the respective changes in the activities of the relevant enzymes. The ratio of 17-hydroxypregnenolone/pregnenolone did not change significantly, while that of dehydroepiandrosterone/17-hydroxypregnenolone grew, indicating increased C17,20 side chain cleavage on the account of C17-hydroxylation both catalyzed by C17-hydroxylase-C17,20-lyase. As was shown by factor analysis, the changes in the maternal steroids were associated with a single common factor, which strongly correlated with all the steroids except dehydroepiandrosterone sulfate. The lack of change in the pregnenolone sulfate/pregnenolone ratio and a marked increase of the ratio dehydroepiandrosterone sulfate to unconjugated dehydroepiandrosterone indicate a different means of formation of both steroid sulfates. On the basis of these data, a scheme of steroid biosynthesis in maternal organism during the critical stages around parturition is proposed.

Neuroactive steroids, their precursors and polar conjugates during parturition and postpartum in maternal blood: 2. Time profiles of pregnanolone isomers.

Time profiles of the pregnanolone isomers epipregnanolone (3 beta-hydroxy-5 beta-pregnan-20-one), allopregnanolone (3 alpha-hydroxy-5 alpha-pregnan-20-one), pregnanolone (3 alpha-hydroxy-5 beta-pregnan-20-one), and isopregnanolone (3 beta-hydroxy-5 alpha-pregnan-20-one) were measured around parturition and in the postpartum period in the serum of 13 and three women with subarachnoidal and epidural analgesia, respectively. In addition, the levels of polar conjugates of all pregnanolone isomers were followed during parturition. GC/MS analysis was used for the measurement of steroid levels. Changes in concentrations of free steroids exhibited a similar pattern, with a fall primarily within the first hour after delivery. The decrease in conjugated steroids was shifted to the interval within the first hour and first day after delivery, and the changes were more pronounced. The time profile of the conjugated/free steroid ratio exhibited a significant decrease within the first hour and the first day after delivery in all of the isomers investigated. A decrease was also observed in the ratio of 3 alpha/3 beta-isomers and 5 alpha/5 beta-isomers around parturition. The possible physiological consequences of the findings are indicated.

Exploratory biochemical data analysis: a comparison of two sample means and diagnostic displays.

The occurrence of acne in women with hyperandrogenemia is well known; a question remains, however, as to whether a further positive relationship can be detected between the intensity of acne and the levels of testosterone, androgen precursors and sex hormone binding globulin (SHBG). A procedure of interactive data analysis extracting relevant information from original data was applied. Exploratory data analysis (EDA) identifies basic statistical features and patterns of data using a variety of diagnostic displays. The need for this step is particularly acute in biochemical and clinical data, the distribution of which is mostly non-Gaussian and often corrupted by the outliers. The omission of EDA can lead to incorrect results and false conclusions. In the EDA (i) several graphical tools for summarizing data are applied, (ii) the peculiarities of a sample distribution are investigated, (iii) a construction of distribution is carried out, (iv) a graphical comparison of the sample distribution with selected theoretical distributions is employed. The proposed procedure is illustrated by typical case study in the evaluation of differences between mean values of serum levels of testosterone, androgen precursors and SHBG in a group of patients with mild and severe forms of acne. A knowledge of the interval estimate of the mean value in both groups enables their comparison at the chosen probability level. As will be apparent from the evaluation of inter-group SHBG differences, an incorrect approach to the determination of group mean values could result in a complete misinterpretation of the data. The results indicate that androgens are not significantly related to the intensity of acne, and that SHBG is higher in patients with more severe forms of acne.

Analysis of large and small samples of biochemical and clinical data.

Statistical software often offers a list of various descriptive statistics of location and scale, but rarely selects an efficient estimate that is statistically adequate for an actual univariate sample. The sample interval estimate for a specified degree of uncertainty seems to be more meaningful if it covers an unknown value of the population parameter. The concept of an interval estimate in medicine is then used for medical decision-making. The proposed methodology, which uses the S-Plus algorithm for biochemical, biological and clinical data analysis contains the following steps: (i) Exploratory data analysis identifies basic statistical features and patterns of the data, the distributions of which are mostly non-normal, non-homogeneous and often corrupted by outliers. (ii) Sample assumptions about data, independence of sample elements, normality and homogeneity are examined. (iii) Power transformation and the Box-Cox transformation to improve sample symmetry and stabilize the spread. (iv) Classical and robust statistics for both large (n>30) and medium-sized samples (15

Transformation in the PC-aided biochemical data analysis.

Data transformations enable expression of original data in a new scale, more suitable for data analysis. In computer-aided interactive analysis of biochemical and clinical data an exploratory data analysis often finds that the sample distribution is systematically skewed or does not accept a sample homogeneity. Under such circumstances the original data should be transformed. The power transformation and the Box-Cox transformation improve sample symmetry and also stabilize variance. Both the Hines-Hines selection graph and the plot of logarithm of a maximum likelihood function allow selection of an optimum transformation parameter. The proposed procedure of data transformation in univariate data analysis is illustrated on a determination of 17-hydroxypregnenolone in umbilical blood of a population of newborns. Lower levels of free 5-ene steroids in umbilical blood and elevated levels of 5-ene steroid sulfates indicate a congenital sex-specific placental sulfatase insufficiency. After examination of statistical assumptions by diagnostic plots of an exploratory data analysis the best estimate of a mean value of 17-hydroxypregnenolone is derived.

Thermodynamic dissociation constants of isocaine, physostigmine and pilocarpine by regression analysis of potentiometric data.

Concentration and mixed dissociation constant(s) of three drug acids, H(J)L, isocaine, physostigmine and pilocarpine, at various ionic strengths, I, in the range 0.03-0.81 and 25 degrees C have been determined with the use of regression analysis of potentiometric titration data when common parameter, pK(a), and group parameters E'(0), L(0), and H(T) are simultaneously refined. Internal calibration of the glass electrode cell in the concentration scale [H(+)] performed during titration was used. The estimate of ill-conditioned group parameters has a great influence on a systematic error in estimated pK(a) and therefore it makes the computational strategy important. As more group parameters are refined and a better fit achieved, a more reliable estimate of dissociation constants results. The thermodynamic dissociation constant, pK(a)(T), an ill-conditioned ion-size parameter, å, and the salting-out coefficient, C, were estimated by non-linear regression of {pK(a), I} data and an extended Debye-Hückel equation. The goodness-of-fit test based on regression diagnostics is a measure of the reliability of parameters, and proves that reliable estimates for isocaine pK(a)(T)(=)8.96(1), å=8(3) A and C=0.50(3) at 25 degrees C, for physostigmine pK(a)(T)(=)8.07(3), å=19(26) A and C=0.64(3) at 25 degrees C, and for pilocarpine pK(a)(T)(=)7.00(1), å=7(1) A and C=0.53(2) at 25 degrees C were found.

Reliability of protonation constants of SNAZOXS oligomers.

Concentration protonation constants of variously protonated of oligomers of sulphoazoxine SNAZOXS were determined by regression analysis of potentiometric titration curves. The group and common parameters were estimated using different computational strategies of three regression programs, MINIQUAD, MIQUV and PSEQUAD. ANOVA proved that six various computational strategies of three regression programs have no significant influence on reliability of protonation constants estimated in comparison with the reproducibility of the titration. Chemical model of protonation equilibria L(2)H(5-), L(2)H(4-)(2), L(2)H(3-)(3), and L(2)H(2-)(4) and reaction scheme of oligomers protonation for SNAZOXS was found.

Multiparametric curve fitting XIV. Modus operandi of the least-squares algorithm MINOPT.

Hybrid least-squares algorithm MINOPT for a nonlinear regression is introduced. MINOPT from CHEMSTAT package combines fast convergence of the Gauss-Newton method in a vicinity of minimum with good convergence of gradient methods for location far from a minimum. Quality of minimization and an accuracy of parameter estimates for six selected models are examined and compared with different derivative least-squares methods of five commercial regression packages.

Multiparametric curve fitting XV statistical analysis and goodness-of-fit test by the least-squares algorithm MINOPT.

Estimation of nonlinear regression quality leads to examination of quality of parameter estimates, a degree of fit, a prediction ability of model proposed and quality of experimental data. Statistical analysis serves for computation of confidence intervals of parameters and confidence bands, the bias of parameters and bias of residuals. Goodness-of-fit test examines classical residuals using various diagnostics and identifies influential points. Mentioned topics of nonlinear model building and testing contained in MINOPT program from CHEMSTAT package are illustrated.

Multiparametric curve fitting-XIII Reliability of formation constants determined by analysis of potentiometric titration data.

The formation (protonation) constants log K(i), of the acid H(j)L are determined by regression analysis of potentiometric titration data when common parameters (log K(i), i = 1,..., j) and group parameters (E(0)', L(0), H(T)) are refined. The influence of three kinds of error on the protonation constants has been investigated: error from the strategy of minimization, random error, and error from uncertain estimates of group parameters. An analysis of variance of the log K(i), matrix was made for 7 identical titrations and 8 computational strategies, or of 7 identical titrations and 8 different options of group parameters to be refined. The influence of the standard potential E(0) of the glass-electrode cell on the systematic error in log K is greater than that of the acid concentration (L(0)) or the concentration of titrant used (H(T)). The ill-conditioned group parameters should be refined together with the common parameters (K(i)), otherwise the estimates of log K(i), are not accurate enough. Two ways of calibrating the glass electrode cell were compared. Internal calibration (performed during titration) was more accurate than external calibration done separately. Of the programs tested ESAB and ACBA are the most powerful because they permit refinement of group parameters and internal calibration. Citric acid was chosen as model substance.

Multiparametric curve fitting-XII Resolution capability of two programs for analysing multicomponent spectra, squad(84) and psequad(83).

The resolving power of multicomponent spectra analysis and the computation reliability of the stability constants and molar absorptivities determined for nine variously protonated anions of three sulphonephthaleins and an impurity, by analysis of data for a mixture by programs SQUAD(84) and PSEQUAD(83), has been examined by use of synthetic and experimentally measured spectra containing severely overlapping spectral bands. The model mixture of Bromocresol Green, Phenol Red, Thymol Blue and azoxine as impurity, with five yellow, three blue and one red species in the pH range from 2 to 10, was used to examine the influence of precision of spectral data and of use of the spectra of the individual components, on the precision and accuracy of the estimated parameters when the chemical model is known. An efficient computation strategy has been found and both programs were shown to lead to the same values and reliability of the parametric estimates. Of the various diagnostics considered, the goodness-of-fit achieved is used as the criterion of whether the parameters found adequately represent the data.

Multiparametric curve fitting--X: a structural classification of programs for analysing multicomponent spectra and their use in equilibrium-model determination.

A functional structure-classification of programs for analysis of spectra elucidates their efficiency for determination of the stoichiometric indices, stability constants and molar absorptivities of complex species. SQUAD (84) introduces new functional units for (i) determination of the number of light-absorbing species, (ii) a rigorous fitness test, (iii) plotting three-dimensional graphs of a paraboloid minimum response-surface as a function of two selected parameters, and a graph of the fitted absorbance response-plane, (iv) simultaneous estimation of stoichiometric indices and stability constants, (v) simulation of an absorbance matrix data by loading with random errors related to the instrumental variance of the absorbance. A guide to experimental procedure and computational strategy for chemical model determination is given and nine diagnostic tools useful in finding the number of species present and their stoichiometry and stability constants by regression analysis of spectra are tested, by use of literature data.

Multiparametric curve fitting-XI: POLET computer program for estimation of formation constants and stoichiometric indices from normalized potentiometric data.

The FORTRAN computer program POLET(84) analyses a set of normalized potentiometric titration curves to find a chemical model, i.e., the number of species present and their stoichiometry, and to determine the corresponding stability constants log beta(pqrs) and unknown stoichiometric indices p, q, r, and s of up to quaternary M(p)L(q)Y(r)H(s) complexes. The program belongs to the ABLET family, based on the LETAG subroutine, and can use an algorithmic and/or heuristic minimization strategy, or a beneficial combination of both. The data, a set of potentiometric titration curves plotted as volume and potential, are converted into normalized variables (formation function, pH) and then a computer-assisted search for a chemical model by POLET(84) is applied. The procedure for efficient application of POLET(84) in an equilibrium analysis is described and the program is validated by use of literature and simulated data. The reliability of the chemical model and its parameters is established by the degree-of-fit achieved, and the closeness of the stoichiometric indices to integral values.

Multiparametric curve fitting-IX Simultaneous regression estimation of stoichiometry and stability constants of complexes.

A chemical model (i.e., the number of complexes, their stoichiometry and stability constants with molar absorptivities) in solution equilibria may be established by (i) the trial-and-error method in which stability constants are estimated for an assumed set of complexes in the mixture and a fitness test is used to resolve a choice of plausible models to find the true one; (ii) the simultaneous estimation of the stoichiometry and stability constants for species divided into "certain" species for which the parameters beta(pqr), (p, q, r) are known and held constant, and "uncertain" species with unknown parameters which are determined by regression analysis. The interdependence of stability constants and particular sets of stoichiometric indices requires that the computational strategy should be chosen carefully for each particular case. The benefits and limitations of both approaches are compared by means of examples of potentiometric titration data analysis by the POLET(84) program and of spectrophotometric data analysis by the SQUAD(84) program. A strategy for efficient computation is suggested.

Multiparametric curve fitting--VIII: The reliability of dissociation constants estimated by analysis of absorbance-pH curves.

The program SPOPT estimates stability constants beta(pqr) and molar absorptivities epsilon(pqr) of all light-absorbing species M(p)L(q)H(r) by analysis of the absorbance-concentration (or absorbance-pH) curve. The program DCMINUIT estimates dissociation constants and molar absorptivities of protonated species. Both programs have been tested and compared with DCLET and LETAGROP-SPEFO for analysis of the overlapping equilibria of a triprotic acid. Computer plots of the residual-square-sum function are used to test the conditioning of parameters. Two approaches are made to formulation of the mathematical model, and several optimization algorithms are tested to find a reliable minimization procedure. The accuracy of ill-conditioned parameters is shown to be dependent on the precision of the absorbance measurements. General rules for investigation of A -pH curves are recommended.

Multiparametric curve fitting-VII Determination of the number of complex species by factor analysis of potentiometric data.

The number of complex species in solution may be determined by a computer-assisted factor analysis of a set of potentiometric titration curves, by finding the rank of the normalized data matrix. An application of the program SPECIES is demonstrated for some examples of titration data. The method is limited in that it can discriminate only between species with differing degrees of polymerization.