Changes in metabolic and muscle damage indicators following a single bout of jump training on stair versus at level.

UNLABELLED: We hypothesized that stair-jump exercise would induce less muscle damage and greater acute metabolic responses than level-jumps. METHODS: Trained males executed 100 unilateral jumps on stairs with one leg, and at level with the other leg, with two weeks hiatus. Maximal isometric voluntary torque (MVC) and rate of torque development (RTD)in the quadriceps, and unilateral vertical jump height (VJ) were determined in the trained leg at pre-exercise,immediately at post- (IP), 24 h and 48 h after exercise. Serum creatine kinase (CK) level and delayed onset muscle soreness (DOMS) were evaluated at pre-exercise, 24 h and 48 h. Acute lactate and heart rate responses were also measured. RESULTS: Lactate and heart rate at IP increased similarly under the two conditions. CK was elevated and MVC was depressed while RTD and VJ remained unchanged at 24 h in both types of training. MVC recovered at 48 h only after stair-jump exercise. DOMS developed only after level-jumps. Except DOMS, no effects of condition were found in any other variables. CONCLUSIONS: We conclude that vigorous stair-jump exercise highly stresses the aerobic and the anaerobic energy system, and it preserves power and rapid torque generating ability 24 h after exercise. Stair-jump could be one alternative exercise to prevent muscle soreness.

Direct effects of chemoradiotherapy following esophagectomy.

BACKGROUND: Esophageal cancer is a major cause of morbidity and mortality, but despite continuing research, few effective therapies have been identified. In recent years, surgical resection following chemoradiotherapy has been associated with improved survival in several clinical models. AIM: In a prospective, observational study, we evaluated the direct effects of chemoradiotherapy on postoperative mortality, morbidity, and inflammatory response in patients following esophagectomy. METHODS: The study cohort was divided into two groups: the first group received preoperative chemoradiotherapy, while the second group had surgical intervention without prior treatment. Nutritional status was evaluated for the members of both patient groups at various time points. RESULTS: Preoperative chemoradiotherapy did not influence morbidity or organ function, and the postoperative inflammatory response did not show immunosuppressive side effects directly after surgery. CONCLUSION: Preoperative chemoradiotherapy does not improve postoperative organ function, inflammatory response or nutritional status in the patients. These findings may help to improve outcome in patients with esophageal cancer in the future.

Selective association of endogenous ouabain with subclinical organ damage in treated hypertensive patients.

According to previous studies endogenous ouabain (EO) closely correlates with high blood pressure, congestive heart failure and kidney disease in humans. Our aims were to analyse associations between plasma, urinary EO level and various markers of cardiovascular damage in treated hypertensive patients. Forty-one adult patients with hypertension and/or diabetes mellitus (DM) and/or chronic kidney disease (CKD) were studied. We assessed plasma and urinary EO, pro-brain natriuretic peptide and catecholamines, profile of ambulatory blood pressure monitor and cardiovascular status by echocardiography and echo-tracking. The highest level of plasma EO (19.7±9.5 pmol l⁻¹) was measured in hypertensive patients with DM and CKD. The nighttime mean arterial blood pressure independently correlated with the level of plasma EO (P=0.004), while independent predictor of the ß-stiffness of carotid artery was the urinary EO (P=0.011). Elevated level of EO was associated with nighttime blood pressure and subclinical organ damage in treated hypertensive patients, suggesting possible role of EO in the pathogenesis of impaired diurnal blood pressure rhythm and arterial stiffness.

Effect of glutamine in patients with esophagus resection.

UNLABELLED: Glutamine is the most abundant amino-acid in the extra- and intracellular compartments of the human body, which accounts for over 50% of its free amino-acid content. Utilization of glutamine peptides is explicitly useful, resulting in a decrease in the number of postoperative infectious complications, period of hospitalization, and therapeutic costs. This article aims to study the effects of glutamine on systemic inflammatory response, morbidity, and mortality after esophagectomy. A prospective, randomized, double-blind, and controlled trial was used. Following sealed-envelope block randomization, the patients were divided into two groups. Members of the glutamine group (group G) received glutamine (Dipeptiven, Fresenius) as continuous infusion for 6 hours at 0.5 g/kg for 3 days prior to, and 7 days following surgery; while patients of the control group were given placebo. We examined 30 patients in group G, and 25 patients as controls. In both patient groups, the levels of total protein, albumin, pre-albumin, retinol binding protein, transferrin, transferring-saturation, C-reactive protein, procalcitonin, lymphocte, Interleukin-6, Interleukin-8, tumor necrosis factor alpha, and serum lactate were determined prior to surgery (t(0)), directly after surgery (t(u)), following surgery on day 1 (t(1)), day 2 (t(2)), and day 7 (t(7)). For statistical analysis Mann-Whitney U test and chi-square test were used. There was no significant difference between the two groups regarding age, male/female ratio, and SAPS II scores. Intensive care unit morbidity and mortality was similar in both groups (group G: 24 survivors/6 nonsurvivors; CONTROL: 17 survivors/8 nonsurvivors; P= 0.607). Daily Multiple Organ Dysfunction Score did not differ significantly between the two groups. The observed inflammatory markers followed the pattern we described without significant difference. Based on our study, the glutamine supplementation that we used had no influence on morbidity, mortality, or postoperative inflammatory response after esophagectomy.

Microalbuminuria in inflammatory bowel diseases using immunoturbidimetry and high-performance liquid chromatography.

BACKGROUND AND STUDY AIMS: To measure urinary albumin excretion using immunoturbidimetry (IT) and high-performance liquid chromatography (HPLC) in inflammatory bowel diseases. PATIENTS AND METHODS: A cross-sectional study was carried out on 60 selected patients with Crohn's disease (CD), 57 with ulcerative colitis (UC) and 22 healthy volunteers, as controls. Urinary albumin excretion was measured by IT and HPLC, and albumin-creatinine ratio was calculated. This ratio was compared in patients with active and inactive CD and UC and in healthy volunteers. RESULTS: Patients with CD and UC had higher albumin-creatinine ratio compared to controls using both IT and HPLC (p < 0.05). We measured higher albumin-creatinine ratio in patients with active compared to inactive CD (p < 0.05). Albuminuria did not correlate with disease duration of CD or UC, but patients with more extended CD according to the Montreal classification had higher HPLC-albumin-creatinine ratio. In CD, we found a significant correlation between HPLC-albumin-creatinine ratio and some inflammatory markers i.e. white blood cells (p < 0.05) and erythrocyte sedimentation rate (p < 0.05). In UC, there was no significant correlation between HPLC-albumin-creatinine ratio and the above markers of systemic inflammation or activity of UC. Albumin-creatinine ratio measured by HPLC was higher in both active and inactive CD and UC groups than albumin-creatinine ratio measured by IT. Using a receiver operating characteristics curve analysis, in case of HPLC-albumin-creatinine ratio cut-off values of the activity of CD were 2.46 mg/mmol for men, 5.30 mg/mmol for women. CONCLUSIONS: HPLC-urinary albumin-creatinine ratio is associated with the clinical and laboratory disease activity indices in CD, but not in UC. Using HPLC we found a more sensitive method compared to IT to measure albuminuria that would be a sensitive activity marker in CD.

Investigation of abiogenic stress-induced alterations in the level of secondary metabolites in poppy plants (Papaver somniferum L.).

We aimed to understand the effects of water stress on the alkaloid production in various developmental stages of poppy plants and the effect of stress on the alkaloids content in the capsules. Three stages of the life cycle of Papaver somniferum L. were selected in our studies: Rosette, Flowering and Lancing developmental stages. Four types of water conditions were examined: Control, Withdrawal of Water, 50% Water Supply and Inundation. The morphological monitoring, results of Relative Water Content and proline content were used as indicators of stress. The result of the measurements in poppy leaves show that the secondary metabolites dramatically respond to these stress conditions. The constant water supply was beneficial for the accumulation of alkaloids in the capsules.

Single dose of acetylsalicylic acid in patients with Type 2 diabetes mellitus and/or chronic renal failure ameliorates anaemia by decreasing the rate of neocytolysis.

BACKGROUND: Anaemia in diabetes mellitus (DM) and/or chronic renal failure (CRF) may be caused by a decreased production of erythropoietin (EPO), EPO resistance, and by the lysis of the young circulating red blood cells (neocytolysis) induced by subclinical inflammation and low EPO level. Aims of this study were to detect EPO resistance in patients with DM and/or CRF and to prove, that acetylsalicylic acid (ASA) is able to improve the haemopoietic status by decreasing neocytolysis. METHODS: In a cross-sectional study, three groups of selected patients (patients with DM; patients with DM+CRF; patients with CRF without DM, n=15 each) and a group of controls (non-diabetic, nonazotemic subjects, n = 10) were compared. In the intervention part of the study, the effect of a single dose of 1 gram ASA on neocytolysis was investigated in a subgroup of these patients. RESULTS: Despite the similar EPO level (p = 1.000), all three patient groups had lower haemoglobin and haematocrit than control persons (p < 0.05 in all cases). Patients with DM+CRF had lower haemoglobin than patients with DM or CRF alone (p < 0.05). Single dose of ASA induced a fast increase in serum EPO level, a concomitant rise of the Rtc number and rate, red blood cell count, haematocrit and haemoglobin p < 0.01 for each). These changes were accompanied by a marked decrease in serum lactate dehydrogenase activity (p < 0.01). CONCLUSIONS: DM and CRF may induce erythropoietin resistance. In these patients, ASA treatment increases serum EPO level. The higher EPO level and the anti-inflammatory effect of ASA may decrease neocytolysis.

Insulin-induced peroxynitrite production in human platelet-rich plasma.

Recent data support the possible role of nitric oxide (NO*) in the development of insulin signalling. The aim of this study was to examine the effect of insulin on NO* production by platelets. The chemiluminescence of platelet-rich plasma prepared from the blood of healthy volunteers was measured in the presence of luminol. Indirect detection of NO* by luminol is possible in the form of peroxynitrite produced in the reaction of NO* with a superoxide free radical. Luminol oxidation induced by hydroxyl free radical and lipid peroxidation was prevented by 150 micromol/l of desferrioxamine mesylate. Insulin, in the range of 0.084-840 nmol/l, induced a concentration-dependent increase in chemiluminescence, which was inhibited both by the competitive antagonist of the NO* synthase enzyme. N(omega)-nitro-L-arginine methyl ester (at concentrations of 2.0-4.0 mmol/l, P<0.001), and by the elimination of superoxide free radicals using superoxide dismutase (72-144 IU/ml, P<0.001). In conclusion, we assume that the insulin-induced increase in chemiluminescence of platelet-rich plasma was due to increased production of NO* and superoxide free radicals forming peroxynitrite. The data are consistent with production of peroxynitrite from human platelets under insulin stimulation.

[Inflammatory markers after surgical treatment of esophageal tumors]. / Gyulladásos markerek szintjének vizsgálata tumor miatti nyelócsómútétek után.

Oesophagectomies carry the risk of postoperative sepsis and mortality. The aim of this study was to evaluate the course of microalbuminuria, serum procalcitonin and C-reactive protein levels following oesophagectomies. Twenty one patients undergoing elective oesophagectomy were studied. Serum procalcitonin and C-reactive protein levels were determined on arrival on the intensive care unit (t0) and then daily (t24, t48, t72). Microalbuminuria (expressed as urine albumin:creatinine ratio, mg/mmol) was measured before (tpre), and after surgery (t0, t6, t24, t48, t72). For statistical analysis Wilcoxon test was used. The clinical course of the patients studied was uneventful during the first 72 hours as monitored by daily Multiple Organ Dysfunction Scores. Preoperative microalbuminuria levels were normal (< 10 mg/mmol). Levels at t0 increased significantly but then (t6-24) they returned to normal. Serum procalcitonin (normal: < 0.5 ng/ml) at t0 was slightly elevated and by t24 it increased significantly (median: 2.7 ng/ml, p < 0.05) and remained high for the rest of the study: t48-72. C-reactive protein was normal at t0 (< 10 mg/l) and by t24 it increased dramatically (up to 10-20 times to the normal value) until t48. At t72 it decreased, but still remained in the abnormal range. This study found, that the surgical insult resulted a significant increase in microalbuminuria, serum procalcitonin and C-reactive protein levels. However, the changes were not accompanied by the clinical signs of sepsis or multiple organ dysfunction in the early postoperative period following oesophagectomies.

Replacement of ATP with ADP affects the dynamic and conformational properties of actin monomer.

The effect of the replacement of ATP with ADP on the conformational and dynamic properties of the actin monomer was investigated, by means of electron paramagnetic resonance (EPR) and fluorescence spectroscopic methods. The measurement of the ATP concentration during these experiments provided the opportunity to estimate the time dependence of ADP-Mg-G-actin concentration in the samples. According to the results of the fluorescence resonance energy transfer experiments, the Gln-41 and Cys-374 residues are closer to each other in the ADP-Mg-G-actin than in the ATP-Mg-G-actin. The fluorescence resonance energy transfer efficiency increased simultaneously with the ADP-G-actin concentration and reached its maximum value within 30 min at 20 degrees C. The EPR data indicate the presence of an ADP-Mg-G-actin population that can be characterized by an increased rotational correlation time, which is similar to the one observed in actin filaments, and exists only transiently. We suggest that the conformational transitions, which were reflected by our EPR data, were coupled with the transient appearance of short actin oligomers during the nucleotide exchange. Besides these relatively fast conformational changes, there is a slower conformational transition that could be detected several hours after the initiation of the nucleotide exchange.

ATP and integrity of human red blood cells.

In spite of the well known significance of ATP in the energy dependent life processes, the role of ATP in maintaining cellular integrity is poorly understood. A possible model for studying ATP dependent life processes is to monitor the kinetics of changes seen intra/extracellularly during ATP depletion. In our model system anticoagulated human whole blood was incubated at different temperatures to reduce intracellular ATP without addition of any chemicals. The red blood cells in their own plasma were incubated for several days at 4 degrees C or at 37 degrees C, and ATP, glucose, K+, Na+, hemoglobin, water content, mean corpuscular volume (MCV), pH and Ca2+ were analyzed in time-sequences. All the examined parameters remained practically unchanged at 4 degrees C, while at 37 degrees C total ATP and glucose decreased parallel and after a transient increase of MCV, the water content of red blood cells decreased. As the actual ATP fell below 10% of the initial ATP content (at 48 h), the release of potassium sharply increased. Release of hemoglobin started only after 96 hours of incubation. Maximums of changes of the examined parameters were found at different time intervals. The maximal speed of concentration changes for glucose was found at 12-24 hours of incubation and at 24-36 hours for ATP, at 48-60 hours for K+(-)Na+ and after 96 hours for hemoglobin.

Maintenance of ions, proteins and water in lens fiber cells before and after treatment with non-ionic detergents.

If the plasma membrane and its associated transport proteins are solely responsible for maintenance of the asymmetric solute distribution then disruption of the plasma membrane would quickly lead to the symmetric distribution of all unattached inorganic ions between the cell and the extracellular environment. To test this hypothesis fresh pig lenses were incubated in Hanks' balanced salt solution in either absence or presence of non-ionic detergents (0.2% Triton X-100 or 0.2% Brij 58). Both detergents caused permeabilization of every lens fiber cell as shown by electron microscopy. The flux kinetics of K+, Mg2+, Na+, Ca2+, water and protein out of and into the permeabilized lens fiber cells was measured. Triton X-100 caused a faster flux rate of all solutes than did Brij 58. The Triton X-100 induced flux of solutes and water was associated with a decrease in lens ATP. Incubation of untreated lenses in solutions of different osmotic pressures at 0 degree C demonstrated that the major fraction of lens water was osmotically unresponsive. Thus the asymmetric distribution of solutes in lens fiber cells is dependent on an intact plasma membrane and on a co-operative ATP-dependent association between K+, Mg2+, water and cytomatrix proteins.

Release of potassium, lipids, and proteins from nonionic detergent treated chicken red blood cells.

The plasma membrane of erythrocytes, as of other cells, is thought to act as the barrier responsible for maintaining intracellular gradients of most ions and small molecular species between the cell and its environment. Controlled application of the nonionic detergent Brij 58 effectively opened the erythrocyte plasma membrane, as judged by electron microscopy and lipid mobilization, but the cytoplasm maintained much of its integrity for about 30 min. Release of K+ correlated well with release of protein into the surrounding medium. The results demonstrate that permeabilization of the erythrocyte plasma membrane does not result in an instantaneous equilibration of small ions, such as K+, between the cell and its environment. A comparison was made between erythrocytes treated with Brij 58 and Triton X-100. The lipid and protein solubilizing actions of Triton X-100 were not as easily separable in time as those of Brij 58. The results of treatment of the erythrocytes with different types of nonionic detergents suggest that the membranolytic and cytoplasmic protein destabilizing actions of nonionic detergents correspond with their hydrophilic-lipophilic balance numbers (HLB values).

Transient cytoplasmic vacuolization in cultured normal and neoplastic cells treated with low molecular weight human serum ultrafiltrate: is out inside?

Human embryonal fibroblasts (HEF) and epithelial carcinoma cells (HEp-2) were exposed to low molecular weight (LMW) fractions (M(R) < 50-kDa) of human serum, generated by ultrafiltration. A prominent change in cytoplasmic morphology was detected by microscopic analysis of both cell types. Within 1-3 h of incubation an extensive vacuolization was induced, which phenotype was maintained for 6-12 h in the presence of the LMW filtrate in a cell type dependent manner, followed by a spontaneous reconstitution of the original cytoplasmic morphology by 24 h. Reversibility occurred by incubation in medium without filtered serum fractions. Transmission electron microscopy revealed vesicles without continuous bordering membrane or electron dense interna and demonstrated vacuoles in close contact with filamentous structures. Taken together the light- and electron microscopic characteristics of the observed vacuolization, potential interpretations of our results are discussed in the context of general cytoplasmic organization. Rearrangement of the cytoplasmic protein-water system as a consequence of an inductive effect of serum filtrates is proposed.

Release kinetics of ATP in cells exposed to nonionic detergents.

We have previously shown that the protein binding of intracellular ATP could be examined by monitoring the ATP release kinetics from Triton X-100 and Brij 58 nonionic detergent permeabilized cells. We have now analysed the protein binding of ATP in an isotonic medium using intact and partially ATP depleted Brij 58 treated human erythrocytes. The effects of Triton X-100 below the critical micelle concentration (CMC) was studied in normal and tumorous tissue culture cells and human red blood cells. Our results showed that the protein association of ATP was altered in the partially ATP depleted erythrocytes. Below the CMC value, but above a critical level Triton X-100 treatment was effective in mobilizing the intracellular ATP in both cell types. The ATP release curves were sigmoidal and an 'all or none' type of response was observed, especially in erythrocytes. The use of Triton X-100 (less than CMC) delays the detergent-induced cell decomposition time thus providing a new approach to investigating the physical state of intracellular ATP.

[Low-level chemoluminescence as a marker of ischemic myocardial damage]. / Az alacsonyszintü chemilumineszcencia mint az ischaemiás myocardialis károsodás kifejezöje.

The low level chemiluminescence of intact and ischemic tissue homogenate of rat and dog heart have been investigated with luminometry. The t-butyl hydroperoxide-induced photoemission is increased in ischemic state and strict differences may be determined between the intact and ischemic samples. The light burst is intensiver in the infarct vs. intact region. On the other hand, the subendocardial layers show higher chemiluminescence.

Cholesteryl hemisuccinate's inductive effect on membrane rigidization regarding both, its remodelling of the cells' surface receptor pattern and its decreasing the natural killer susceptibility of K-562 cells.

The relationship of changes in membrane fluidity to natural killer susceptibility of K-562 target cells was investigated. Membrane rigidization was performed by the chemical modulator cholesteryl hemisuccinate. Steady-state fluorescence polarization measurements of the diphenyl hexatriene labelled, modified K-562 cells revealed that cholesteryl hemisuccinate increased the structural order of the hydrophobic region of membranes in a dose dependent way. Investigation of natural killer susceptibility followed by 51Cr release assay indicated that modified cells are less sensitive to natural killer attack. To elucidate whether surface structures such as transferrin and lectin receptors are associated with the altered susceptibility, the surface density of these receptors was followed by (I-125)-transferrin binding assay and quantitative immunofluorescence. We found that the number of transferrin and concanavalin A receptors increased by a factor of 2.44 and 2.00, respectively, whereas that of the wheat germ agglutinin receptor failed to exhibit any changes upon rigidization. From the results we concluded that i the membrane structural order does influence the natural killer susceptibility, ii changes in membrane structural order result in alteration of the number of cell surface transferrin and lectin receptors, iii however, no direct relationship seems to exist between these two events.

Membrane fluidity of trophoblast cells and susceptibility to natural cytotoxicity.

This study examines the relationship between membrane lipid microviscosity and susceptibility of villous trophoblast to lysis by natural cytotoxic cells. Trophoblast-enriched cell suspensions prepared from term human placentae were treated with cholesteryl hemisuccinate (CHS)--a modulator of membrane lipid microviscosity. CHS-treated cells were more susceptible targets for natural lymphocyte cytotoxicity than were untreated controls. In binding experiments, increased binding of lymphocytes to CHS-treated target cells was found. Preincubation with progesterone prevented membrane rigidification by CHS. Progesterone, cortisol, and estriol restored the impaired resistance of CHS-treated trophoblast cells to lysis. We determined microviscosity and progesterone concentration in villous surface membranes, prepared from placentae from idiopathic spontaneous abortions and normal first-trimester pregnancies. An inverse relationship was found between progesterone content and microviscosity of the membranes. Microviscosity of the membranes from abortion placentae was significantly higher (P less than .01) and progesterone concentration was significantly lower (P less than .001) than those in the membranes of normal first trimester placentae.

The relationship of membrane fluidity to growth and nuclear cycle in mitogen stimulated lymphocytes.

Fluorescence polarization (P) and anisotropy (rs) of the diphenyl-hexatriene labeled, stimulated human lymphocytes were followed in relation to cell cycle progression. Fluorescence anisotropy is proportional to the structural order parameter of the apolar lipid region, thus, indicative of the membrane fluidity. Cell cycle progression was defined by monitoring the RNA, protein and DNA synthesis as well as the expression of interleukin 2 and transferrin receptors. Cellular growth in size was also measured. A decrease in P and rs values was seen when the RNA and protein synthesis as well as the density of growth factor receptors and cellular volume increased. At the same time no DNA synthesizing cell was detected. The low P and rs values remained unaltered throughout the cycle, thus proving to be independent from cell proliferation (DNA synthesis) and from further progressive increase in cellular volume. The data indicate that the structural change in the membranes is an early event of cell activation and occurs in the course of G0-G1 transition as part of the growth cycle.