Enhancement of carbohydrates in a methylotrophic yeast

The use of methylotrophic yeasts has been suggested for recycling CO₂ to human food in extended space missions. Since the human diet requires higher carbohydrate levels than those normally found in microbes, attempts were made to increase the levels of storage carbohydrates, principally glycogen, through cultural and genetic methods. The effect of defining cultural conditions for the methylotrophic yeast Hansenula polymorpha resulted in increasing the storage carbohydrate content of the dry weight of the cells from 30 to 46%. During this analysis, a growth requirement for potassium was discovered. Several mutant strains were selected for high glycogen storage on plates and analysed for storage carbohydrate levels in submerged culture. These strains exhibited an additional 4–16% increase in the levels of storage carbohydrates over the parent strain in stationary phase. One strain was also able to store excess carbohydrate during exponential growth at levels 10% above the parent strain. Through a combination of cultural control and genetic modification, carbohydrate levels in this yeast were raised to 60% of the cell dry weight. Through additional genetic selection these levels are likely to be increased even further.     

Structure of the Ca2+ pump of sarcoplasmic reticulum: a view along the lipid bilayer at 9-A resolution.

We have used multilamellar crystals of the ATP-driven calcium pump from sarcoplasmic reticulum to address the structural effects of calcium binding to the enzyme. They are stacks of disk-shaped two-dimensional crystals. A density map projected along the lipid bilayer was obtained at 9-A resolution by frozen-hydrated electron microscopy. Although only in projection, much more details of the structure were revealed than previously available, especially in the transmembrane region. Quantitative comparison was made with the model obtained from the tubular crystals of this enzyme formed in the absence of calcium. Unexpectedly large differences in conformation were found, particularly in the cytoplasmic domain.     

Acoustic myography reflects force changes during dynamic concentric and eccentric contractions of the human biceps brachii muscle.

The relationship between acoustic myography (AMG), electromyography (EMG) and force during submaximal dynamic contractions was examined in the biceps brachii muscles of eight healthy males (aged 17-26 years). Different weights were lifted and lowered at a constant speed, using a wall pulley system, to perform concentric and eccentric contractions, respectively. Integrated AMG (iAMG) and integrated EMG (iEMG) activity both increased linearly with force during concentric (iAMG r = 0.94; iEMG r = 0.99) and eccentric (iAMG r = 0.90; iEMG r = 0.94) contractions. The slopes of the concentric regression lines were significantly different from the eccentric slopes (P less than 0.01) for both iAMG and iEMG with concentric contractions showing greater levels of activity. The results indicated that AMG can be used to detect changes in force during dynamic contractions which has important implications for the use of AMG in rehabilitation. The differences in iAMG activity between concentric and eccentric contractions are discussed in relationship to the origin of the AMG signal.     

The ATP-binding site of Ca(2+)-ATPase revealed by electron image analysis.

The location of the ATP-binding site of a P-type ion pump, Ca(2+)-ATPase from rabbit sarcoplasmic reticulum, was examined by cryoelectron microscopy. A nonhydrolyzable analog of ATP, beta, gamma-bidentate chromium (III) complex of ATP (CrATP), was used to stabilize the enzyme in the Ca(2+)-occluded state. Tubular crystals were then induced by vanadate in the presence of EGTA, keeping CrATP bound to the enzyme. The three-dimensional structures of the crystals were determined at 14 A resolution by cryoelectron microscopy and helical image analysis. Statistical comparison of the structures with and without CrATP showed clear and significant differences at the groove proposed previously as the ATP-binding pocket.     

Cemental annulation enhancement: a technique for age determination in man

Cemental annulations are easily countable in teeth from animals that have an exaggerated regular change of food intake from season to season. Although present in human teeth, cemental annulations are not always easy to count. A method for preparing human teeth for evaluation involving collection, identification, measuring, sectioning, cleaning, acid etching, staining, and mounting is reported. Sections 100-microns thick were stained with cresyl fast violet as a stain of choice and were photographed using standard light microscopic techniques as well as Nomarsky interference microscopy. Countability of annulations from photographic enlargements was evaluated.     

Measurement of anterior tibial muscle size using real-time ultrasound imaging.

Cross-sectional images of the anterior tibial muscle group were obtained using real-time ultrasound scanning in 17 normal women. From photographs taken of the images, the cross-sectional area (CSA) and two linear measurements of muscle cross-section were determined. A measurement of the shortest distance of the muscle depth was termed DS, and a measurement of the longest distance through the muscle group was termed DL. Both linear dimensions showed a positive correlation with CSA and the best correlations were obtained when the dimensions were squared or combined (DS x DL). The correlation values were: CSA vs DS2, r = 0.9; CSA vs DL2, r = 0.75 and CSA vs DS x DL, r = 0.88. An approximate value for CSA could be calculated from DS2 by the equation 2 x DS2 + 1. A shape ratio, obtained by dividing DL by DS, was consistent within the group [mean 2.1 (SD 0.2)] and characterised the muscle geometrically. The CSA of repeated scans was assessed for repeatability between-days and between-scans by analysis of variance and the coefficient of variation (CV) calculated. Areas were repeatable between-days (CV 6.5%) and between-scans (CV 3.6%). Linear dimensions of the anterior tibial muscle group reflected CSA and their potential for assessing changes in muscle size with atrophy and hypertrophy have yet to be established.