ACUTE EFFECTS OF MOVEMENT VELOCITY ON BLOOD LACTATE AND GROWTH HORMONE RESPONSES AFTER ECCENTRIC BENCH PRESS EXERCISE IN RESISTANCE-TRAINED MEN

This study aimed to compare the effects of different velocities of eccentric muscle actions on acute blood lactate and serum growth hormone (GH) concentrations following free weight bench press exercises performed by resistance-trained men. Sixteen healthy men were divided into two groups: slow eccentric velocity (SEV; n = 8) and fast eccentric velocity (FEV; n = 8). Both groups performed four sets of eight eccentric repetitions at an intensity of 70% of their one repetition maximum eccentric (1RMecc) test, with 2-minute rest intervals between sets. The eccentric velocity was controlled to 3 seconds per range of motion for SEV and 0.5 seconds for the FEV group. There was a significant difference (P < 0.001) in the kinetics of blood lactate removal (at 3, 6, 9, 15, and 20 min) and higher mean values for peak blood lactate (P = 0.001) for the SEV group (9.1 ± 0.5 mM) compared to the FEV group (6.1 ± 0.4 mM). Additionally, serum GH concentrations were significantly higher (P < 0.001) at 15 minutes after bench press exercise in the SEV group (1.7 ± 0.6 ng · mL⁻¹) relative to the FEV group (0.1 ± 0.0 ng · mL⁻¹). In conclusion, the velocity of eccentric muscle action influences acute responses following bench press exercises performed by resistance-trained men using a slow velocity resulting in a greater metabolic stress and hormone response.     

Thermodynamic and Kinetic Insights into Stop Codon Recognition by Release Factor 1

Stop codon recognition is a crucial event during translation termination and is performed by class I release factors (RF1 and RF2 in bacterial cells). Recent crystal structures showed that stop codon recognition is achieved mainly through a network of hydrogen bonds and stacking interactions between the stop codon and conserved residues in domain II of RF1/RF2. Additionally, previous studies suggested that recognition of stop codons is coupled to proper positioning of RF1 on the ribosome, which is essential for triggering peptide release. In this study we mutated four conserved residues in Escherichia coli RF1 (Gln185, Arg186, Thr190, and Thr198) that are proposed to be critical for discriminating stop codons from sense codons. Our thermodynamic and kinetic analysis of these RF1 mutants showed that the mutations inhibited the binding of RF1 to the ribosome. However, the mutations in RF1 did not affect the rate of peptide release, showing that imperfect recognition of the stop codon does not affect the proper positioning of RF1 on the ribosome.     

Changes in membrane biophysical properties induced by sphingomyelinase depend on the sphingolipid N-acyl chain

Ceramide (Cer) is involved in the regulation of several cellular processes by mechanisms that depend on Cer-induced changes on membrane biophysical properties. Accumulating evidence shows that Cers with different N-acyl chain composition differentially impact cell physiology, which may in part be due to specific alterations in membrane biophysical properties. We now address how the sphingolipid (SL) N-acyl chain affects membrane properties in cultured human embryonic kidney cells by overexpressing different Cer synthases (CerSs). Our results show an increase in the order of cellular membranes in CerS2-transfected cells caused by the enrichment in very long acyl chain SLs. Formation of Cer upon treatment of cells with bacterial sphingomyelinase promoted sequential changes in the properties of the membranes: after an initial increase in the order of the fluid plasma membrane, reorganization into domains with gel-like properties whose characteristics are dependent on the acyl chain structure of the Cer was observed. Moreover, the extent of alterations of membrane properties correlates with the amount of Cer formed. These data reinforce the significance of Cer-induced changes on membrane biophysical properties as a likely molecular mechanism by which different acyl chain Cers exert their specific biological actions.     

TAM receptor ligands in lupus: Protein S but not Gas6 levels reflect disease activity in systemic lupus erythematosus

Introduction  

The TAM (tyro 3, axl, mer) kinases are key regulators of innate immunity and are important in the phagocytosis of apoptotic cells. Gas6 and protein S are ligands for these TAM kinases and bind to phosphatidyl serine residues exposed during apoptosis. In animal models, absence of TAM kinases is associated with lupus-like disease. To test whether human systemic lupus erythematosus (SLE) patients might have deficient levels of TAM ligands, we measured Gas 6 and protein S levels in SLE.     

A quantitative electrochemiluminescence assay for Clostridium perfringens alpha toxin

Described is a rapid direct sandwich format electrochemiluminescence assay for identifying and assaying Clostridium perfringens alpha toxin. Biotinylated antibodies to C. perfringens alpha toxin bound to streptavidin paramagnetic beads specifically immunoadsorbed soluble sample alpha toxin which subsequently selectively immunoadsorbed ruthenium (Ru)-labeled detection antibodies. The ruthenium chelate of detection antibodies chemically reacted in the presence of tripropylamine and upon electronic stimulation emitted photons (electrochemiluminescence) that were detected by the photodiode of the detector. Elevated toxin concentrations increased toxin immunoadsorption and the specific immunoadsorption of Ru-labeled antibodies to alpha toxin, which resulted in increased dose-dependent electrochemiluminescent signals. The standardized assay was rapid (single 2.5-h coincubation of all reagents), required no wash steps, and had a sensitivity of about 1 ng/ml of toxin. The assay had excellent accuracy and precision and was validated in buffer, serum, and urine with no apparent matrix effects.     

Deciduous woodland conservation under heavy deer browsing on Devils Tower National Monument, USA

Native deciduous woodlands comprise <5% of the vegetation in the Black Hills and northern plains of the USA and appear to be in decline. Because deciduous woodlands comprise 18% of the land cover on Devils Tower National Monument (DTNM), this natural area is potentially important for the conservation of these plant communities. Lack of tree regeneration and accumulation of dead wood suggest that the native deciduous woodlands on DTNM may die out under persistent, heavy deer (Odocoileus virginianus, O. hemionus) browsing. We monitored vegetation changes for seven years (1989–1996) inside and outside 6 deer exclosures on DTNM to determine the resilience of native woodland communities when protected from browsing, and to parameterize models that evaluate specific fencing scenarios for managing woodland communities. The herbaceous woodland understory was not directly, but indirectly affected by deer browsing because graminoid biomass declined as tall shrubs recovered. Tall shrubs, dominated by Prunus spp. showed the most rapid and consistent recovery across sites. Models of shrub growth indicated that rotational fencing for short durations (10 years) produced the highest long-term increase in tall-shrub densities and the greatest evenness among size classes. Because deciduous woodlands currently are resilient and occur in small patches, fencing may provide a practical management alternative to deer culling programs.     

Territory surveillance and prey management: Wolves keep track of space and time

Identifying behavioral mechanisms that underlie observed movement patterns is difficult when animals employ sophisticated cognitive‐based strategies. Such strategies may arise when timing of return visits is important, for instance to allow for resource renewal or territorial patrolling. We fitted spatially explicit random‐walk models to GPS movement data of six wolves (Canis lupus; Linnaeus, 1758) from Alberta, Canada to investigate the importance of the following: (1) territorial surveillance likely related to renewal of scent marks along territorial edges, to reduce intraspecific risk among packs, and (2) delay in return to recently hunted areas, which may be related to anti‐predator responses of prey under varying prey densities. The movement models incorporated the spatiotemporal variable “time since last visit,” which acts as a wolf's memory index of its travel history and is integrated into the movement decision along with its position in relation to territory boundaries and information on local prey densities. We used a model selection framework to test hypotheses about the combined importance of these variables in wolf movement strategies. Time‐dependent movement for territory surveillance was supported by all wolf movement tracks. Wolves generally avoided territory edges, but this avoidance was reduced as time since last visit increased. Time‐dependent prey management was weak except in one wolf. This wolf selected locations with longer time since last visit and lower prey density, which led to a longer delay in revisiting high prey density sites. Our study shows that we can use spatially explicit random walks to identify behavioral strategies that merge environmental information and explicit spatiotemporal information on past movements (i.e., “when” and “where”) to make movement decisions. The approach allows us to better understand cognition‐based movement in relation to dynamic environments and resources.     

Hemodynamic effects of different preparations of stroma free hemolysates in the isolated perfused rat kidney

We have examined the effects of Stroma Free Hemolysate (SFH) solutions in the isolated perfused rat kidney. Three types of SFH, stored for 6 to 8 months at 4 degrees C, were tested: 1) unmodified, 2) glyoxalated and lightly cross linked and 3) pyridoxalated and polymerized. All three SFH solutions, added to the perfusate at a concentration of approximately 420 mg/100ml, increased renal vascular resistance (RVR) and reduced glomerular filtration rate (GFR). Unmodified, glyoxalated and lightly cross linked and pyridoxalated polymerized SFH resulted in a rise in RVR of 55%, 38% and 33% respectively and a fall in GFR of 42%, 57% and 83% respectively. In order to determine whether storage had altered the effect of SFH on renal function, one of the forms of SFH (glyoxalated and lightly cross linked) was studied only 4-6 weeks after preparation. While this preparation caused an increase in RVR of 41% it did not alter GFR; filtration fraction (FF) rose. However, after further storage of this preparation for 6-7 months, the solution resulted in a marked decrease in GFR of 47% as well as a rise in RVR of 23%. We conclude that three different SFH preparations resulted in marked vasoconstriction and reductions in GFR. These deleterious effects on renal hemodynamics were noted at a concentration of hemoglobin well below that necessary to effectively improve oxygen content. Storage of the SFH solutions may cause or contribute to their effects on renal function. SFH solutions intended for use as blood substitutes should be tested for vasoconstrictor activity.