Biological activity and receptor binding of human prointerleukin-1 beta and subpeptides

We report here that the human interleukin-1 beta precursor (proIL-1 beta) protein as well as several interleukin-1 beta (IL-1 beta) subpeptides bind cellular receptors specifically and exhibit biological activity by stimulating proliferation of helper T-cells. IL-1 beta polypeptides have been synthesized by in vitro translation of mRNAs transcribed from plasmid vectors containing the bacteriophage SP6 promoter joined to the complete IL-1 beta cDNA or to deletion constructs. The quantity of IL-1 beta in vitro translation products was increased significantly by replacing the cognate IL-1 beta untranslated leader sequence with a 37-nucleotide plant viral untranslated leader. Translation of chimeric mRNAs followed by direct bioactivity assay demonstrated that mature IL-1 beta-(117-269), proIL-1 beta-(1-269), and peptide IL-1-(71-269) were all biologically active. Specific binding to cellular receptors was observed with these three IL-1 beta molecules; moreover, several peptides with minimal biological activity also bound receptor specifically. The biological activity and receptor binding properties of the IL-1 beta proteins reported here contrast with those described by Mosley et al. (Mosley, B., Urdal, D. L., Prickett, K. S., Larsen, A., Cosman, D., Conlon, P. J., Gillis, S., and Dower, S. K. (1987) J. Biol. Chem. 262, 2941-2944; Mosley, B., Dower, S. K., Gillis, S., and Cosman, D. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 4572-4576), who reported that proIL-1 beta-(1-269) had no biological activity and does not bind receptor. Our results indicate that proIL-1 beta is active at a relatively high concentration, and analysis of the proIL-1 beta-(1-269) and IL-1-(71-269) bioactivity data suggests a possible relationship with membrane-bound IL-1.     

Seasonal fluctuations of selected physiological characteristics of elite alpine skiers

The effects of heavy resistance training and jumping exercise were examined during the 1989–1990 season in 12 international level alpine skiers. The athletes were tested before, during, immediately after training and during the period off training (June, July, October 1989, April 1990). Their mechanical behaviour was investigated using firstly squat jumps performed without (SJ) or with low extra loads (20 kg, SJ_20kg) and high extra loads (equivalent to body mass on the shoulders, SJ_bm) and secondly 15–30 s continuous jumping. These tests allowed the assessment of explosive dynamic strength production (SJ and SJ_20kg), slow dynamic strength (SJ_bm) and maximal mechanical power (continuous jumping). The training adopted resulted in specific changes in neuromuscular performance; in fact all the variables studied showed a significant improvement (P<0.01) from the beginning compared to the end of training. The range of improvement was between 55.4% (SJ_bm) and 12.5% (average power during 15-s continuous jumping). The enhancement of SJ had become significant by July. Surprisingly, even when no strength or jumping training was performed during the competition period (November-April), no deterioration in the neuromuscular performance was observed, there being no significant difference between the test values obtained in October 1989 and April 1990. It was concluded that the demanding competition programme of alpine skiers may provide a training stimulus adequate to maintain the neuromuscular improvement induced by training throughout the competition season.     

Low Density Lipoproteins Promote Unstable Calcium Handling Accompanied by Reduced SERCA2 and Connexin-40 Expression in Cardiomyocytes

The damaging effects of high plasma levels of cholesterol in the cardiovascular system are widely known, but little attention has been paid to direct effects on cardiomyocyte function. We therefore aimed at testing the hypothesis that Low Density Lipoprotein (LDL) cholesterol affects calcium dynamics and signal propagation in cultured atrial myocytes. For this purpose, mRNA and protein expression levels were determined by real time PCR and western blot analysis, respectively, and intracellular calcium was visualized in fluo-4 loaded atrial HL-1 myocyte cultures subjected to field stimulation. At low stimulation frequencies all cultures had uniform calcium transients at all tested LDL concentrations. However, 500 µg LDL/mL maximally reduced the calcium transient amplitude by 43% from 0.30±0.04 to 0.17±0.02 (p<0.05). Moreover, LDL-cholesterol dose-dependently increased the fraction of alternating and irregular beat-to-beat responses observed when the stimulation interval was shortened. This effect was linked to a concurrent reduction in SERCA2, RyR2, IP3RI and IP3RII mRNA levels. SERCA2 protein levels were also reduced by 43% at 200 µg LDL/mL (p<0.05) and SR calcium loading was reduced by 38±6% (p<0.001). By contrast, HDL-cholesterol had no significant effect on SERCA expression or SR calcium loading. LDL-cholesterol also slowed the conduction velocity of the calcium signal from 3.2+0.2 mm/s without LDL to 1.7±0.1 mm/s with 500 µg LDL/mL (p<0.05). This coincided with a reduction in Cx40 expression (by 44±3%; p<0.05 for mRNA and by 79±2%; p<0.05 for Cx40 protein at 200 µg/ml LDL) whereas the Cx-43 expression did not significantly change. In conclusion, LDL-cholesterol destabilizes calcium handling in cultured atrial myocytes subjected to rapid pacing by reducing SERCA2 and Cx40 expression and by slowing the conduction velocity of the calcium signal.     

Estimation of Unsteady Aerodynamics in the Wake of a Freely Flying European Starling (Sturnus vulgaris)

Wing flapping is one of the most widespread propulsion methods found in nature; however, the current understanding of the aerodynamics in bird wakes is incomplete. The role of the unsteady motion in the flow and its contribution to the aerodynamics is still an open question. In the current study, the wake of a freely flying European starling has been investigated using long-duration high-speed Particle Image Velocimetry (PIV) in the near wake. Kinematic analysis of the wings and body of the bird has been performed using additional high-speed cameras that recorded the bird movement simultaneously with the PIV measurements. The wake evolution of four complete wingbeats has been characterized through reconstruction of the time-resolved data, and the aerodynamics in the wake have been analyzed in terms of the streamwise forces acting on the bird. The profile drag from classical aerodynamics was found to be positive during most of the wingbeat cycle, yet kinematic images show that the bird does not decelerate. It is shown that unsteady aerodynamics are necessary to satisfy the drag/thrust balance by approximating the unsteady drag term. These findings may shed light on the flight efficiency of birds by providing a partial answer to how they minimize drag during flapping flight.     

Expansion of Islet-Resident Macrophages Leads to Inflammation Affecting β Cell Proliferation and Function in Obesity

1. Download high-res image (213KB)
2. Download full-size image

     

Investigation of the possibility of exceeding the Greenwald density limit during ECRH in T-10

In T-10 experiments, attempts were made to significantly exceed the Greenwald limit $\bar n_{Gr} $ during high-power (P _ab=750 kW) electron-cyclotron resonance heating (ECRH) and gas puffing. Formally, the density limit $(\bar n_e )_{\lim } $ exceeding the Greenwald limit $({{(\bar n_e )_{\lim } } \mathord{\left/ {\vphantom {{(\bar n_e )_{\lim } } {\bar n_{Gr} }}} \right. \kern-0em} {\bar n_{Gr} }} = 1.8)$ was achieved for q _L=8.2. However, as q _L decreased, the ratio ${{(\bar n_e )_{\lim } } \mathord{\left/ {\vphantom {{(\bar n_e )_{\lim } } {\bar n_{Gr} }}} \right. \kern-0em} {\bar n_{Gr} }}$ also decreased, approaching unity at q _L≈3. It was suggested that the “current radius” (i.e., the radius of the magnetic surface enclosing the bulk of the plasma current I _p), rather than the limiter radius, was the parameter governing the value of $(\bar n_e )_{\lim } $ . In the ECRH experiments, no substantial degradation of plasma confinement was observed up to $\bar n_e \sim 0.9(\bar n_e )_{\lim } $ regardless of the ratio ${{(\bar n_e )_{\lim } } \mathord{\left/ {\vphantom {{(\bar n_e )_{\lim } } {\bar n_{Gr} }}} \right. \kern-0em} {\bar n_{Gr} }}$ . In different scenarios of the density growth up to $(\bar n_e )_{\lim } $ , two types of disruptions related to the density limit were observed.     

Prediction of microbial growth rate versus biomass yield by a metabolic network with kinetic parameters

Identifying the factors that determine microbial growth rate under various environmental and genetic conditions is a major challenge of systems biology. While current genome-scale metabolic modeling approaches enable us to successfully predict a variety of metabolic phenotypes, including maximal biomass yield, the prediction of actual growth rate is a long standing goal. This gap stems from strictly relying on data regarding reaction stoichiometry and directionality, without accounting for enzyme kinetic considerations. Here we present a novel metabolic network-based approach, MetabOlic Modeling with ENzyme kineTics (MOMENT), which predicts metabolic flux rate and growth rate by utilizing prior data on enzyme turnover rates and enzyme molecular weights, without requiring measurements of nutrient uptake rates. The method is based on an identified design principle of metabolism in which enzymes catalyzing high flux reactions across different media tend to be more efficient in terms of having higher turnover numbers. Extending upon previous attempts to utilize kinetic data in genome-scale metabolic modeling, our approach takes into account the requirement for specific enzyme concentrations for catalyzing predicted metabolic flux rates, considering isozymes, protein complexes, and multi-functional enzymes. MOMENT is shown to significantly improve the prediction accuracy of various metabolic phenotypes in E. coli, including intracellular flux rates and changes in gene expression levels under different growth rates. Most importantly, MOMENT is shown to predict growth rates of E. coli under a diverse set of media that are correlated with experimental measurements, markedly improving upon existing state-of-the art stoichiometric modeling approaches. These results support the view that a physiological bound on cellular enzyme concentrations is a key factor that determines microbial growth rate.     

Efficacy and safety of propofol sedation during urgent upper gastrointestinal endoscopy--a prospective study

The aim of this study was to investigate both the efficacy and safety of sedation with propofol during urgent therapeutic gastroscopy in patients with upper gastrointestinal bleeding. This prospective study included a total of 110 patients. Propofol was administered intravenously at the starting dose of 1 mg/kg body weight and was followed by repeated doses. Oxygen saturation and heart rate were monitored by pulse oxymetry. The mean dose of propofol administered was 161 +/- 49 mg. Urgent upper GI endoscopy under propofol sedation was successful in 98% of cases. Endoscopists rated the sedation as good in 83.6%, satisfactory in 14.5%, and poor in 1.8% of patients. Potentially harmful drop in oxygen saturation below 85% was observed in 5.5% of patients, whereas a temporary drop in heart rate below 50 beats/min was observed in 11.8%, not requiring any intervention. Almost 93% of patients could not remember the beginning or the end of the intervention. This data demonstrates that sedation with propofol is suitable for use in patients with upper gastrointestinal bleeding undergoing urgent endoscopy.     

Benign epithelial gastric polyps--frequency,location, and age and sex distribution

Prospective investigation has been undertaken with the aim to study the frequency, location and age and sex distribution of various histological types of benign gastric epithelial polyps. Histological type--adenomatous, hyperplastic and fundic gland polyps--was diagnosed on the basis of at least three histological samples taken from the polyp. Biopsy samples were also taken from the antrum and the body of the stomach so that gastritis could be graded and classified, and the presence of H. pylori could be determined by histology. All 6,700 patients, who had undergone upper gastrointestinal endoscopy in a one-year period, participated in this study. Among them 42 benign gastric epithelial polyp were found in 31 patients: adenomatous gastric polyps in 7 patients, hyperplastic gastric polyp in 21 and fundic gland polyp in 3 patients. All patients with hyperplastic polyps had chronic active superficial gastritis, whereas most of the patients with adenomatous polyps had a chronic atrophic gastritis with high prevalence of intestinal metaplasia. Among 21 patients with hyperplastic gastric polyps, 16 (76%) patients were positive for H. pylori infection in contrast to only 2 patients (29%) with adenomatous gastric polyps and 1 patient (33%) with fundic gland polyp. Presented data indicates that hyperplastic gastric polyps are the most common and they are associated with the presence of chronic active superficial gastritis and concomitant H. pylori infection. Adenomatous polyps are rarer and they tend to be associated with chronic atrophic gastritis and intestinal metaplasia. Fundic gland polyp is the rarest type of gastric polyps.