Study of the carbohydrate part of yeast acid phosphatase

It has been found that the carbohydrate part of acid phosphatase from yeast Saccharomyces cerevisiae consists of 16 N-glycosidically linked carbohydrate chains containing from 14 to about 150 mannose units. The presence of very small amounts of O-glycosidically linked chains was indicated. Acetolysis studies pointed to a high similarity in the structure of acid phosphatase and mannan carbohydrate chains. A new method is described for cross-linking of acid phosphatase specifically via carbohydrate chains. The possibility to cross-link the enzyme subunits intramolecularly is in accordance with the suggestion that carbohydrate chains play a role in subunit associations.     

Corrosion behaviour of the Co-Cr-Mo dental alloy in solutions of different composition and different pH values

The purpose of this study was to examine how Co-Cr-Mo dental alloy behaves in the solutions of different pH value and different composition over a relatively long period of time. Co-Cr-Mo dental alloy was exposed in vitro to either simulated saliva (phosphate buffer pH 6.0), a highly acidic medium resembling the extreme conditions in the oral cavity (phosphate buffer pH 3.5), and in lactic acid at pH which occurs under the dental plaque (lactic acid pH 3.5). The alloy samples were immersed in these three solution for 1, 2, 3, 4, 5, 6, 7, 14, 21, and 30 days. Solutions were analysed with the ICP-AES. The analysis showed that during one month cobalt (Co), chromium (Cr), iron (Fe), zinc (Zn) and nickel (Ni) ions had been released from tested samples in all three solutions. The results of this study indicate that the leaching of the Co, Cr, Fe, Zn and Ni ions in the solution was dependent both upon the nature of the solution in which the alloy was immersed and the duration of the immersion (p < 0.001).     

Cation-pi interaction in a folded polypeptide.

Cationic and aromatic side chains from protein residues interact to stabilize tertiary structure. The stabilization energy originates in part from electrostatic attraction between the cation, and regions of high electron density in pi-orbitals of the aromatic group, leading to the name cation-pi interaction. The lysine and tyrosine containing peptide, N-acetyl-Pro-Pro-Lys-Tyr-Asp-Lys-NH(2), has near uv CD characteristic of tyrosine in a structured environment. Nuclear Overhauser effect (NOE), coupling constant, and ring current chemical shift constraints obtained with (1)H NMR confirm that the peptide (t6p) folds. Simulated annealing consistent with all NMR constraints produces a 40-structure ensemble for t6p with potential energies within one standard deviation of the lowest value observed. Calculated binding energies indicate that cation-pi and cation-phenolic OH interactions exists between the Lys3 and Tyr4 side chains in most of the structures. The t6p peptide in solution is a model for these interactions in a protein. A perturbing electric field from the cationic ground state charge intermingles the excited states of the aromatic group. This intermingling effect may provide a cation-pi signature effect in the tyrosine spectroscopy. The absorption and CD for the lowest energy electronic transitions of the tyrosine phenol were computed for the ensemble. Red-shifted peak energy and hypochromicity in the absorbance band, and decreasing rotational strength, correlates with increasing binding energy of the complex indicating the cation-pi spectroscopic signature. The ensemble average spectroscopic signature effects in t6p are small and in agreement with observation.     

Regulation of T-type calcium channels in the peripheral pain pathway

Recent evidence strongly suggests that both central and peripheral T-type Ca(2+) channels enhance somatic and visceral nociceptive inputs, as well as that regulation of T-type Ca(2+) channel function can result in significant changes of pain threshold in a variety of animal models. Therefore, T-type Ca(2+) channels in peripheral and central pain pathways, although previously unrecognized, may have great importance as targets for developing new therapies against pain. This is particularly critical in cases in which currently available treatments are limited due to serious side effects or are not consistently effective (e.g., chronic neuropathic pain). In this review, we summarize recent studies of the regulation of T-type channels in peripheral sensory neurons by means of redox agents and neuroactive steroids, as well as studies of the function of these channels in the pathophysiology of neuropathic pain.     

The histone chaperone Asf1 increases the rate of histone eviction at the yeast PHO5 and PHO8 promoters

Eukaryotic gene expression starts off from a largely obstructive chromatin substrate that has to be rendered accessible by regulated mechanisms of chromatin remodeling. The yeast PHO5 promoter is a well known example for the contribution of positioned nucleosomes to gene repression and for extensive chromatin remodeling in the course of gene induction. Recently, the mechanism of this remodeling process was shown to lead to the disassembly of promoter nucleosomes and the eviction of the constituent histones in trans. This finding called for a histone acceptor in trans and thus made histone chaperones likely to be involved in this process. In this study we have shown that the histone chaperone Asf1 increases the rate of histone eviction at the PHO5 promoter. In the absence of Asf1 histone eviction is delayed, but the final outcome of the chromatin transition is not affected. The same is true for the coregulated PHO8 promoter where induction also leads to histone eviction and where the rate of histone loss is reduced in asf1 strains as well, although less severely. Importantly, the final extent of chromatin remodeling is not affected. We have also presented evidence that Asf1 and the SWI/SNF chromatin remodeling complex work in distinct parallel but functionally overlapping pathways, i.e. they both contribute toward the same outcome without being mutually strictly dependent.     

Amino-acid Type Identification in 15N-HSQC Spectra by Combinatorial Selective 15N-labelling

The efficiency of cell-free protein synthesis combined with combinatorial selective ¹⁵N-labelling provides a method for the rapid assignment of ¹⁵N-HSQC cross-peaks to the 19 different non-proline amino-acid types from five ¹⁵N-HSQC spectra. This strategy was explored with two different constructs of the C-terminal domain V of the τ subunit of the Escherichia coli DNA polymerase III holoenzyme, τ_C16 and τ_C14. Since each of the five ¹⁵N-HSQC spectra contained only about one third of the cross-peaks present in uniformly labelled samples, spectral overlap was much reduced. All ¹⁵N-HSQC cross-peaks of the backbone amides could be assigned to the correct amino-acid type. Availability of the residue-type information greatly assisted the evaluation of the changes in chemical shifts observed for corresponding residues in τ_C16 vs. those in τ_C14, and the analysis of the structure and mobility of the C-terminal residues present in τ_C16 but not in τ_C14.     

Kinematics and kinetics of unanticipated misstep conditions: Femoral fracture implications in the elderly

Most hip fractures are thought to occur after falling during everyday activities. We speculated that hip fractures might also occur because of excessive loading of the hip joint during an unexpected misstep consequently leading to a fall. The aims of this study were to explore the kinematics and kinetics of the lower extremity joints during missteps as compared with regular stepping, as well as to compare the magnitude of forces acting upon the hip joint with the threshold forces expected to fracture the hip. Fourteen healthy adults performed two forward steps on a 17.8 cm high platform under the following four conditions: forward with and without vision, as well as a misstep (the box for the final step unexpectedly removed without participant awareness), and regular stepping down with eyes open. The results revealed no differences between stepping forward with and without vision. When compared with both stepping forward and regular stepping down, the misstep revealed altered joint positions accompanied by increased forces and moments acting upon the hip joint. For example, the peak vertical proximal thigh segment force was 3.05±0.55 BW vs. 1.23±0.14 BW and 0.91±0.09 BW (p<.001; misstep vs. regular stepping down and stepping forward, respectively), while the proximal thigh segment moment in frontal plane was 1.39±0.70 Nm/kg vs. 0.18±0.32 Nm/kg of adduction and 0.16±0.19 Nm/kg of abduction (p<.001). When compared with the literature data, the forces during misstep were within the range of those forces that could result in hip fractures in the elderly. Therefore, it may be possible for the elderly to experience hip/proximal femur fractures during missteps prior to falling.     

Development of anthropometric and physical performance profiles of young elite male soccer players: a longitudinal study

The purpose of the present longitudinal study was to explore distinctive anthropometric and physical performance characteristics of young soccer players between the age of 11 and 14 and to reveal the performance at the age of 11, which contributes to the later success. Male players of the best national male squads of the 'cadet league' (14 years of age; n = 26) were annually tested starting from the age of 11 for body size and composition, flexibility, power, coordination, and agility. Randomly selected untrained but physically active age-matched boys (n = 63) were also tested over 4 consecutive years. The results revealed no difference between 2 groups regarding the body size and composition (p > 0.05). The differences in flexibility emerged only at the later age, whereas the differences regarding the explosive power (as assessed by various jumps) were moderate and partly inconsistent. The most prominent advantage of the soccer players over the control subjects during the entire tested age period appeared to be movement agility and coordination (p < 0.01). Therefore, the explosive muscle power and, in particular, the agility and coordination characterize elite soccer players of 11-14 years of age but not the body size and body composition. In addition, the agility and coordination could be among the crucial factors of future success in 11-year-old players and, therefore, should be used for early selection.