Interaction patterns of methoprene-tolerant and germ cell-expressed Drosophila JH receptors suggest significant differences in their functioning.

Methoprene-tolerant (Met) and germ cell-expressed (Gce) proteins were shown to be juvenile hormone (JH) receptors of Drosophila melanogaster with partially redundant functions. We raised the question of where the functional differentiation of paralogs comes from. Therefore, we tested Met and Gce interaction patterns with selected partners. In this study, we showed the ability of Gce and its C-terminus (GceC) to interact with 14-3-3 in the absence of JH. In contrast, Met or Met C-terminus (MetC) interactions with 14-3-3 were not observed. We also performed a detailed structural analysis of Met/Gce interactions with the nuclear receptor fushi tarazu factor-1 (Ftz-F1) ligand-binding domain. We showed that GceC comprising an Ftz-F1-binding site and full-length protein interacts with Ftz-F1. In contrast to Gce, only MetC (not full-length Met) can interact with Ftz-F1 in the absence of JH. We propose that the described differences result from the distinct tertiary structure and accessibility of binding sites in the full-length Met/Gce. Moreover, we hypothesize that each interacting partner can force disordered MetC and GceC to change the structure in a partner-specific manner. The observed interactions seem to determine the subcellular localization of Met/Gce by forcing their translocation between the nucleus and the cytoplasm, which may affect the activity of the proteins. The presented differences between Met and Gce can be crucial for their functional differentiation during D. melanogaster development and indicate Gce as a more universal and more active paralog. It is consistent with the theory indicating gce as an ancestor gene.

Multielemental analysis of 18 essential and toxic elements in amniotic fluid samples by ICP-MS: Full procedure validation and estimation of measurement uncertainty.

Amniotic fluid is the substantial factor in the development of an embryo and fetus due to the fact that water and solutes contained in it penetrate the fetal membranes in an hydrostatic and osmotic way as well as being swallowed by the fetus. Elemental composition of amniotic fluid influences the growth and health of the fetus, therefore, an analysis of amniotic fluid is important because the results would indicate abnormal levels of minerals or toxic elements. Inductively coupled plasma mass spectroscopy (ICP-MS) is often used for determination of trace and ultra-trace level elements in a wide range of matrices including biological samples because of its unique analytical capabilities. In the case of trace and ultra-trace level analysis detailed characteristics of analytical procedure as well as properties of the analytical result are particularly important. The purpose of this study was to develop a new analytical procedure for multielemental analysis of 18 elements (Al, As, Ba, Ca, Cd, Co, Cr, Cu, Mg, Mn, Ni, Pb, Sb, Se, Sr, U, V and Zn) in amniotic fluid samples using ICP-MS. Dynamic reaction cell (DRC) with two reaction gases, ammonia and oxygen, was involved in the experiment to eliminate spectral interferences. Detailed validation was conducted using 3 certified reference mterials (CRMs) and real amniotic fluid samples collected from patients. Repeatability for all analyzed analytes was found to range from 0.70% to 8.0% and for intermediate precision results varied from 1.3% to 15%. Trueness expressed as recovery ranged from 80% to 125%. Traceability was assured through the analyses of CRMs. Uncertainty of the results was also evaluated using single-laboratory validation approach. The obtained expanded uncertainty (U) results for CRMs, expressed as a percentage of the concentration of an analyte, were found to be between 8.3% for V and 45% for Cd. Standard uncertainty of the precision was found to have a greater influence on the combined standard uncertainty than on trueness factor.

Mutagenesis of mitochondrial DNA in Fuchs endothelial corneal dystrophy.

Fuchs endothelial corneal dystrophy (FECD) is an age-related, slowly progressive disease, which may lead to loss of vision resulting from apoptosis of corneal endothelial (CE) cells, dysfunction of Descemet membrane (DM) and corneal edema. A growing body of evidence suggests that oxidative stress may play a major role in the pathogenesis of FECD and that mitochondria of CE cells are its main target. Mitochondrial DNA (mtDNA) is particularly prone to oxidative stress and changes in mtDNA were reported in FECD patients. In the present work we studied mtDNA damage and repair, mtDNA copy number, and the 4977bp common deletion in mtDNA in DM cells and peripheral blood lymphocytes (PBLs) isolated from FECD patients. PBLs from 35 FECD patients and 32 controls were challenged for 10min with hydrogen peroxide at 20µM and then left in a fresh medium for 3h, resulting in a decrease in mtDNA copy number in both groups. Damage to mtDNA was not fully repaired after 3h and the extent of remaining lesions was significantly higher in the patients than the controls. We observed a higher copy number and an increased extent of mtDNA damage as well as a higher ratio of the common 4977bp deletion in DM cells of FECD patients than the controls. Our results confirm that mutagenesis of mtDNA may be involved in FECD pathogenesis and disturbance in mtDNA sensitivity to damaging agent as well as changes in mtDNA damage repair along with alternations in mtDNA copy number may underline this involvement.

Resting energy expenditure and gut microbiota in obese and normal weight subjects.

OBJECTIVES: It is suggested that gut microbiota play a role in the pathogenesis of obesity enhancing energy utilization from digested food. The influence of gut microbiota on resting energy expenditure (REE) has not been evaluated yet. AIM: The aim of the study is to assess the composition on gut microbiota and its association with REE in obese and normal weight subjects. SUBJECTS AND METHODS: REE measurement and semi-quantitative analysis of gut microbiota composition in aerobic and anaerobic conditions were performed in 50 obese and 30 normal weight subjects without concomitant diseases. RESULTS: A count of bacterial colony was greater in obese than in normal weight subjects. However, the proportion of Bacteroides spp. and Firmicutes was similar in both study groups. A positive correlation between REE (kcal/d) and total bacterial count (r = 0.26, p < 0.05), as well as between REE and the percentage of Firmicutes (r = -0.24, p < 0.05) was found. The multiple regression analysis did not prove an independent impact of total bacterial as well as Bacteroides spp. and Firmicutes counts on REE. CONCLUSIONS: The composition of gut microbiota is not associated with the level of resting energy expenditure. The proportion of Bacteroides and Firmicutes in gut microbiota is not related to body mass.

The influence of a 3-month weight reduction therapy with Orlistat on serum vitamin B12 and folic acid concentration in obese women.

OBJECTIVE: Serum folic acid, but not the vitamin B(12) concentration, was found to be significantly lower in obese subjects than in the control ones. DESIGN: The aim of this study was to examine the levels of serum vitamin B(12) and folic acid in obese women before and after weight reduction therapy with Orlistat in comparison to healthy controls with normal body weight. SUBJECTS: Twenty obese women participated in a 3-month weight reduction therapy. The control group consisted of 20 healthy women. MEASUREMENTS: Body weight and height were measured and BMI was calculated. Body composition was analyzed with the impedance method using a Bodystat analyzer. In all patients before and after 3-month weight reduction therapy, serum concentrations of folic acid and vitamin B(12) were assessed. RESULTS: In obese women, serum concentrations of folic acid and vitamin B(12) did not change significantly after 3-month weight reduction therapy with Orlistat.

Metabolic effects associated with adipose tissue distribution.

Cardiovascular and metabolic risk depends not only on the overall obesity but also fat distribution is more powerfull predictor for risk factors. Adipose tissue produces and secretes a variety of bioactive peptides - adipokines The most recently described adipocyte secretory proteins contribute to the pathogenesis of impaired insulin secretion and insulin resistance, endothelial dysfunction, a proinflammatory state and promote progression of atherosclerosis. This review presents an overview of the adipose tissue secreted proteins (leptin, TNF-alpha, IL-6, adiponectin, resistin, visfatin, ASP, FIAF, MT) role and their regulation in the context of abdominal obesity and the adverse metabolic consequences.

Ligand binding is without effect on complex formation of the ligand binding domain of the ecdysone receptor (EcR).

The ligand-binding domain (LBD) encompassing the C-terminal parts of the D- and the complete E-domains of the ecdysteroid receptor (EcR) fused to Gal4(AD) is present in two high molecular weight complexes (600 and 150 kDa) in yeast extracts according to size exclusion chromatography (Superdex 200 HR 10/30). Hormone binding is mainly associated with 150-kDa complexes. Complex formation is not influenced by hormone, but the ligand stabilizes the complexes at elevated salt concentrations. Mutational analysis of Gal4(AD)-EcR(LBD) revealed that formation of 600-kDa, but not 150-kDa, complexes depends on dimerization mediated by the EcR(LBD). Deletion of helix 12 is without effect. Mutation of K497 in helix 4, known to be essential for comodulator binding, abolishes 600-KDa complexes, but does not interfere with the formation of 150-kDa complexes. In contrast, the DE-domains of USP fused to Gal4(DBD) elute as monomer after elimination of the dimerization capacity of the ligand-binding domains by mutation of P463 in helix 10. The data presented here reveal that the complex formation of ligand-binding domains EcR and USP ligand is different.

Prevalence of obesity in Poland.

Obesity is an important health problem in Poland affecting adolescents and adults. A study of a Lower Silesian population aged 20-40 years (25 400 participants), between 1993 and 2003, showed an increase in obese women from 8.9% to 15.0%, but no change in men (6.3% vs. 6.5%, respectively). Obesity occurs more frequently in girls and women (6% and 15%) compared to boys and men (4.0% and 6.5%, respectively). Estimation of obesity prevalence in various parts of Poland seems to be regionally dependent; the highest prevalence of obesity was observed in eastern regions of Poland.

[Tumor necrosis factor alpha and its soluble receptors in serum of patients with coronary artery disease]. / Czynnik martwicy nowotworów alpha i rozpuszczalne formy jego receptorów w surowicy u chorych na chorobe wiencowa.

TNF-alpha as a pleiotropic, proinflammatory cytokine seems to play a role in the pathogenesis of atherosclerosis and coronary artery disease (CAD). TNF-alpha is binding to two cell surface receptors and its serum activity is modified by soluble forms of these receptors: sTNF-R I and sTNF-R II. The aim of this study was to assess serum concentrations of TNF-alpha, sTNF-R I and sTNF-R II in patients (pts) with CAD. We examined serum concentrations of TNF-alpha, sTNF-R I and sTNF-R II by ELISA in: 45 pts with stable exertional angina (group I); 32 pts with unstable angina (group II) within 6, 24, and 48 h after the chest pain; and 23 pts before and 6, 24, and 48 h after PTCA (group III). The control group (group C) consisted of 20 healthy subjects. We evaluated: clinical state of patients and results of some diagnostic examinations (lipids, ECG, echocardiography, coronary angiography). Mean serum concentrations of TNF-alpha were significantly higher in pts ith CAD (group I: 18.25 +/- 5.5 pg/ml; group II: 17.24 +/- 4.0 pg/ml; group III: 18.70 +/- 0.6 pg/ml; p < 0.001) than in healthy pts (8.31 +/- 1.4 pg/ml). In turn mean serum concentrations of sTNF-R I were significantly higher both in group I (1399.6 +/- 536.3 pg/ml; p < 0.05) and III (1544.0 +/- 391.4 pg/ml; p < 0.01) than in control group (1093.9 +/- 456.9 pg/ml). There were not differences in mean serum concentrations of sTNF-R II. We found no differences between mean serum concentrations of TNF-alpha, sTNF-R I and sTNF-R II either after the chest pain (group II); or before and after PTCA (group III). In group I mean TNF-alpha correlated with serum triglycerides and HDL-cholesterol (r = 0.412 and r = -0.424; p < 0.01); sTNF-R I correlated with LDL-cholesterol (r = -0.309; p < 0.05); and sTNF-R II correlated with total cholesterol and LDL-cholesterol (r = 0.311 and r = 0.316; p < 0.05). The serum concentrations of TNF-alpha are increased in patients with CAD, but this does not reflect the clinical state of patients. In pts with stable angina these increased levels of TNF-alpha may be accompanied with higher concentrations of sTNF-R I--it seems to be the compensatory mechanism in long-term atherosclerosis. Lipid disturbances may influence the cytokines metabolism in pts with CAD.

Neuropeptide Y in obese women during treatment with adrenergic modulation drugs.

BACKGROUND: The aim of the study is the assessment whether weight loss treatment with adrenergic modulation drugs modifies neuropeptide Y (NPY) plasma concentration in obese women. MATERIAL AND METHODS: 13 obese women (BMI 38.3 +/- 4.4) were tested before and subsequently 10 and 20 days after weight loss treatment. The treatment consisted of a very low caloric diet of 400 kcal (1670 kJ) daily combined with ephedrine with caffeine (E + C) or ephedrine with caffeine and yohimbine (E + C + Y) administered for 10 days using the cross-over method. The patients underwent physical examination, including heart rate and blood pressure measurements, spectral heart rate variability (HRV) at rest and after 3 minute handgrip and a 15 minute cycloergometer exercise at 75 W. All the above mentioned tests were carried out thrice in each patient. In 13 obese patients and in 6 control women plasma NPY concentrations were determined by a specific radioimmunoassay using rabbit anti-NPY antiserum and a standard synthetic porcine NPY (Peninsula Lab.). RESULTS: Plasma NPY concentrations were significantly lower in the obese persons compared with the control group. During weight loss treatment with adrenergic modulation drugs no changes in plasma NPY were found at rest and after physical exercise. Also no differences in HRV indices were observed. CONCLUSIONS: 1. Low plasma NPY concentration observed in obesity may be a contraregulatory factor that could prevent further weight increase. 2. Weight reduction treatment did not affect plasma NPY concentration and cardiovascular response to physical exercise. 3. The doses of adrenergic modulation drugs used in our study did not induce any serious side effects, and were so low that no change of plasma NPY concentration and cardiovascular responses were observed at rest.

[P-selectin and E-selectin in serum of patients with coronary artery disease]. / Selektyna P i selektyna E w surowicy krwi chorych na chorobe wiencowa.

UNLABELLED: Evidence for the role of chronic inflammation in atherogenesis has been well documented. Selectins mediate the first step in leukocyte adhesion and may contribute to the pathogenesis of stable and unstable angina. METHODS: The study group consisted of 59 patients (pts) with coronary artery disease (CAD) documented coronarographically: 27 pts with stable exertional angina (group A), 32 pts with unstable angina (group B). 20 healthy persons were the control group (group C). Serum levels of E-selectin and P-selectin were measured by ELISA method both before and after the treadmill ECG stress test (ST) in groups A and C. In group B the measurements were carried out at 6, 24, and 48 hours following an episode of chest pain. RESULTS: There were no differences between the baseline serum levels of selectins as determined in groups A and C. In patients with stable angina, the post-ST concentrations of E-selectin were significantly higher (68.8 +/- 29 ng/ml) in comparison to both baseline (38.7 +/- 15 ng/ml), and group C-values (pre-ST: 35.1 +/- 16; post-ST: 49.9 +/- 15 ng/ml). In unstable patients, serum P-selectin levels were higher when compared to those found in groups A and C (group A: 142.3 +/- 24; group B: 190.1 +/- 99; group K: 136.4 +/- 33 ng/ml). No differences between selectins concentrations were observed at fixed times after an episode of chest pain. CONCLUSIONS: Soluble selectins levels in pts with stable angina are comparable to those of healthy persons. Significant increase of E-selectin concentration as induced by ST may reflect endothelial response to exercise. Patients with unstable angina had elevated levels of P-selectin, which seems to be associated with enhanced platelet and leukocyte activation. The serum levels of selectins may indirectly reflect clinical condition of pts with CAD.

[The QT interval dispersion and ventricular late potential in obese women]. / Ocena dyspersji odstepu QT i wystepowania póznych potencjalów komorowych u kobiet z otyloscia prosta.

UNLABELLED: Obesity is known to be a risk factor of cardiac death, that is associated first of all with cardiac arrhythmias. Increased QT dispersion (QTd) and ventricular late potentials (LP) are measurable indices of ventricular arrhythmias risk. The aim of this study is to asses QTd and LP in women with obesity. 62 obese women (mean BMI 36.7 kg/m2) and 15 apparently healthy subjects (mean BMI 24.5 kg/m2) were included in our study. QTd and correlated QT interval dispersion (QTdc) were calculated from 12-lead ECG. LVM were assessed from echocardiograms. LP were obtained by signal averaging of surface electrocardiograms. RESULTS: QTd, QTdc, LVM were significantly higher in patients. We found LP in six cases and higher QTdc in this subgroup. According to our research, increased QTd in obese women seems to be associated with left ventricular hypertrophy and increased QTd is significantly higher in subjects with LP.

Serum concentrations of TNF-alpha and soluble TNF-alpha receptors in obesity.

OBJECTIVE: To study whether weight reducing treatment modulates serum concentration of TNF-alpha and two soluble TNF-alpha receptors in obese subjects. SUBJECTS AND MEASUREMENTS: Serum concentrations of TNF-alpha and two soluble receptors (sTNF-R1, sTNF-R2), plasma glucose, insulin, total cholesterol, HDL-cholesterol and triglicerides were measured in 27 obese subjects (age 48 +/- 12y, body mass index (BMI): 36 +/- 6 kg/m2) before and after 3 months weight reducing treatment consisted of a diet & 1,000 kcal/day and physical exercises. RESULTS: The mean loss of weight during 3 months' treatment was 9.3 +/- 3.3 kg. The serum concentration of TNF-alpha decreased after weight loss and at the same time both of the receptors (sTNF-R1,sTNF-R2) increased significantly. CONCLUSION: The observed decrease of the serum concentration of TNF-alpha and the increase in both TNF soluble receptors after weight reducing treatment in obese subjects, may be a counter-regulation preventing further weight loss.

[The selected pro- and anti-inflammatory cytokines in the patients with coronary heart disease: preliminary communication]. / Wybrane cytokiny pro- i antyzapalne w surowicy krwi chorych z choroba wiencowa--doniesienie wstepne.

Recent findings suggest that inflammation and cytokines regulation may play a role in the pathogenesis of atherosclerosis and coronary heart disease. The aim of this study was to assess serum concentrations of selected pro- (TNF alpha) and antiinflammatory (IL-10) cytokines in patients with coronary heart disease. We studied 29 patients with coronary heart disease: 14 with stable angina (group I) and 15 with unstable angina (group II). The control group (group K) consisted of 10 healthy subjects. Patients with inflammatory diseases, previous myocardial infarction (last 6 months) and with ECG abnormalities, that would invalidate ST-segment analysis, were excluded from examined groups. We evaluated: clinical state of patients and results of some diagnostic examinations (lipids, ECG, echocardiography, coronary angiography, concomitant diseases). In each patients serum levels of TNF alpha and IL-10 were measured according to the special protocol by ELISA. The mean serum concentrations of TNF alpha and IL-10 were significantly higher in group I (respectively: 18.75 +/- 11.7 pg/ml, 89.0 +/- 114.9 pg/ml) and II (14.21 +/- 5.9 pg/ml, 49.38 +/- 72.9 pg/ml) in comparison to the healthy subjects (9.41 +/- 1.7 pg/ml, 9.69 +/- 4.5 pg/ml). We found positive correlations between mean TNF alpha and IL-10 concentrations in group II (48 hours after last symptom) and between mean TNF alpha concentration and LVM (left ventricular mass), LVMI (left ventricular mass index) in group I. The concentrations of TNF alpha and IL-10 did not correlate with other clinical parameters. The results of our study suggest that serum concentrations of pro- (TNF alpha) and antiinflammatory (IL-10) cytokines may be increased in patients with stable and unstable angina. These increased concentrations do not reflect the clinical state of patients.

[QT interval dispersion and the type of obesity in women]. / Dyspersja odstepu QT a typ otylosci u kobiet.

UNLABELLED: The risk of sudden cardiac death is higher in patients with obesity, particularly in the upper body obesity. The most common cause of cardiac events are serious ventricular arrhythmias. Delayed cardiac repolarization leading to the prolongation of the QT interval is a well characterised precursor of arrhythmias. The QT interval dispersion reflects inhomogeneity of repolarization. The aim of this study was to assess QT interval dispersion (QTd) in obese women and to establish the relationship between obesity and QTd. 62 patients with obesity (group 1) and 15 apparently healthy women (group 2) were included in our study. Obese subjects were distinguished in accordance to the waist to hip ratio (WHR) into three subgroups: 1a--obese women with upper body obesity; 1b--obese women with WHR: 0.75-0.85; 1c--obese women with lower body obesity. A standard 12-lead ECG was performed in each subjects and QTd, QTdc (QT correlated interval dispersion), QTdR (QTd ratio) were calculated. The left ventricular mass (LVM) and left ventricular mass index (LVMI) were obtained from echocardiograms. We found QTd, QTdc, QTdR and LVM, LVMI to be significantly higher in obese women as well as positive correlation between BMI and both LVM, LVMI. QTdR was significantly higher in subgroup 1a compared with subgroup 1c. CONCLUSIONS: 1) QT interval dispersion was increased in obese women compared with healthy subjects, 2) supposedly increased QTd was associated with the type of obesity.

Pharmacokinetics of theophylline in obesity.

Pharmacokinetic analysis of theophylline was performed in 16 obese women before and after 3-week weight-reducing treatment. Decrease of clearance, increase of t1/2, AUC, and MRT were observed. There were no differences between the volume of distribution before and after weight-reducing treatment. Our results suggest that ideal body weight should be used to calculate a loading dose of theophylline for obese patients; weight-reducing treatment may be connected with changes in biotransformation and elimination of theophylline more than with its distribution.

Resting energy expenditure in patients with cirrhosis of the liver measured by indirect calorimetry, anthropometry and bioelectrical impedance analysis.

Energy expenditure was investigated in 15 patients with liver cirrhosis and 20 healthy controls by three methods: indirect calorimetry, anthropometry using the Harris-Benedict equation and bioelectrical impedance analysis. The energy expenditure was expressed in kcal/day, kcal/kg BW/day (BW - body weight), kcal/kg LBM/day (LBM - lean body mass, derived by bioelectrical impedance analysis) or in kcal/m2/day. We did not find statistical differences between values of resting energy expenditure obtained in patients with cirrhosis of the liver and healthy controls whichever method we used. We also did not find statistical differences between values obtained by indirect calorimetry, anthropometry and bioelectrical impedance analysis. There was a significant correlation between indirect calorimetry and anthropometry in both groups. We found significant correlations between indirect calorimetry and anthropometry, and between indirect calorimetry and bioelectrical impedance analysis, in the control group only. We can conclude that (1) resting energy expenditure of patients with cirrhosis of the liver is not changed when compared with healthy controls, and (2) bioelectrical impedance is a useful method to calculate body composition from which energy expenditure is derived; however, it gives an appropriate result only in healthy people, and only approximate values in patients with cirrhosis.

Microbial transformation of isoprenoid systems by means of fungi of Zoophthora genus--microbial transformation of compounds with alpha-campholenic system by means of fungus Zoophthora (Neopandora) sp.

Unsaturated ketones (4 and 10) and epoxides (2-3 and 8-9) were the main products of biotransformation performed by means of Zoophthora (Neopandora) phyllobii. Enantiospecificity of both reactions leading to these compounds strongly depends on the distance of reacting fragment of molecule from the chiral center at C-1'.