The effect of investigators on the breeding success of royal,Eudyptes schlegeli,and rockhopper penguins,E. chrysocome,at Macquarie Island

The impact on reproductive success of investigators studying the breeding biology of royal and rockhopper penguins was assessed. Control and experimental transects were established in a colony of each species and the number of active nests, from egg laying to creche stage, were compared. Experimental nests were those used in breeding biology work, where birds were measured and banded, and nest checks were carried out at least once per week. Control nests were in equivalent locations but birds were not handled, and no contact was made with the nests once breeding had begun. There were no significant differences in the number of active nests between the control and experimental transects (and, therefore, breeding success) in either species. It is concluded that, provided care is taken when working with these species, no impacts on the short-term (up to creche stage, in one breeding season) breeding success of these populations will occur.     

Cucumber mosaic virus genome is encapsidated in alfalfa mosaic virus coat protein expressed in transgenic tobacco plants

The expression of viral coat protein (CP) in transgenic plants has been shown to be very effective in virus plant protection. However, the introduction of CP genes into plants presents the potential risk of the encapsidation of a superinfecting viral genome in the transgenic protein, an event which could change the epidemiology of the disease. To detect the potential heterologous encapsidation of the cucumber mosaic virus (CMV) genome by alfalfa mosaic virus (AIMV) CP expressed in transgenic tobacco plants, a system of immunocapture (IC) and amplification by polymerase chain reaction (PCR) was optimized. This provided high sensitivity and reliable selection of the heterologously encapsidated CMV genome in the presence of natural CMV particles. As little as 2 pg of virus could be detected by immunocapture/polymerase chain reaction (IC/PCR) technique. Evidence for heterologous encapsidation of the CMV genome was found in 11 of the 33 transgenic plants tested two weeks after CMV inoculation. This demonstrates a significant rate of heterologous encapsidation events between two unrelated viruses in transgenic plants. Since CP is involved in the interactions of the virus particle with its vector, the release in the field of such transgenic plants could alter the transmission properties of some important viruses.     

Molecular evolution of voltage-sensitive ion channel genes: on the origins of electrical excitability

We have analyzed nucleic acid and amino acid sequence alignments of a variety of voltage-sensitive ion channels, using several methods for phylogenetic tree reconstruction. Ancient duplications within this family gave rise to three distantly related groups, one consisting of the Na+ and Ca++ channels, another the K+ channels, and a third including the cyclic nucleotide-binding channels. A series of gene duplications produced at least seven mammalian homologues of the Drosophila Shaker K+ channel; clones of only three of these genes are available from all three mammalian species examined (mouse, rat, and human), pointing to specific genes that have yet to be recovered in one or another of these species. The Shaw-related K+ channels and the Na+ channel family have also undergone considerable expansion in mammals, relative to flies. These expansions presumably reflect the needs of the high degree of physiological and neuronal complexity of mammals. Analysis of the separate domains of the four-domain channels (Ca++ and Na+) supports their having evolved by two sequential gene duplications and implies the historical existence of a functional two-domain channel.      

Fat content of expressed breast milk: a case for quality control.

Expressed breast milk used to feed preterm infants is precious and so, despite heterogeneity of composition, all available milk is used. A study of 274 samples of expressed breast milk supplied by preterm mothers and National Childbirth Trust donors showed pronounced variation in fat content as measured by the "creamatocrit" method. This was not due to differences between term and preterm mothers or between transitional and mature milk. The composition was affected by diurnal variation and method of collection. Substantial amounts of fat were also wasted as a result of continuous nasogastric feeding. Several milk samples did not contain enough fat to supply even a fraction of the recommended energy requirements of these infants. Some type of quality control over samples of expressed breast milk is clearly essential. The creamatocrit method is simple and feasible.     

15N nmr spectroscopy. I. Polysarcosine and related sequence polypeptides

The natural abundance ¹⁵N nmr spectra of linear polysarcosine (DP = 35) has been recorded in Me₂SO and H₂O solution. Because of cis/trans isomerization at the peptide bond, a broad signal with several splittings was observed. These splittings appear to reflect the influence of three peptide bonds on a single N atom. The ¹⁵N signals from the sequence polypeptides (β-Ala-Sar-Gly)_n and (β-Ala-Sar-D ,L -Ala)_n also show a cis/trans splitting, as well as chemical shifts which are dependent on the peptide sequence. The tertiary nitrogen of the sarcosyl residue has a T₁ relaxation time which is longer than the T₁ for secondary nitrogens of the other amino acids. The nuclear Overhauser effect is also discussed.     

Rotational flexibility of the human knee due to varus/valgus and axial moments in vivo

Knee ligamentous injuries persist in the sport of Alpine skiing. To better understand the load mechanisms which lead to injury, pure varus/valgus and pure axial moments were applied both singly and in combination to the right knees of six human test subjects. The corresponding relative knee rotations in three degrees of freedom were measured. Knee flexion angles for each test subject were 15 and 60 degrees for the individual moments and 60 degrees for the combination moments. For both knee flexion angles the hip flexion angle was 0 degrees. Leg muscles were quiescent and axial force was minimal during all tests. Tables of data include sample statistics for each of four flexibility parameters in each loading direction. Data were analyzed statistically to test for significant differences in flexibility parameters between the test conditions. In flexing the knee from 15 to 60 degrees, the resulting knee rotations under single moments depended upon flexion angle with varus, valgus, and internal rotations increasing significantly. Also, rotations were different depending on load direction; varus rotation was significantly different and greater than valgus rotation at both flexion angles. Also external rotation was significantly different and greater than internal at 15 degrees flexion, but not at 60 degrees flexion. Coupled rotations under single moments were also observed. Applying pure varus/valgus moments resulted in coupled external/internal rotations which were inconsistent and hence not significant. Applying pure axial moments resulted in consistent and hence significant varus/valgus rotations; an external axial moment induced varus rotation and an internal axial moment induced valgus rotation. For combination moments, varus/valgus rotations decreased significantly from those rotations at similar load levels in the single moment studies. Also, a varus moment significantly increased external rotation and a valgus moment significantly decreased internal rotation. These differences indicate significant interaction between corresponding load combinations. These results suggest that load interaction is a potentially important phenomenon in knee injury mechanics.     

Structural and functional analysis of the human neutrophil 1-15 antigen, an Mr 65,000 to 70,000 activation-associated membrane proteinase

Previous studies with the anti-neutrophil/antichymotrypsin mAb 1-15 have identified an activation-associated, chymotrypsin-like activity within the membrane fraction of isolated human neutrophils (PMN). In the present study, the molecular and biochemical characteristics of mAb 1-15 Ag/proteinase were determined. On casein/acrylamide sizing gels, PMN membrane preparations were found to contain an Mr 58,000 to 84,000 band of Ca2(+)-dependent proteinase activity. Reducing and nonreducing SDS-PAGE of mAb 1-15-affinity-purified membrane proteins demonstrated specific recovery of an enzymatically active Mr 65,000 to 70,000 chymotrypsin-like Ag. The presence of a distinct membrane serine esterase of isoelectric point 6.3/Mr 65,000 to 70,000 was confirmed in active site-labeling experiments with the serine proteinase inhibitor [3H]diisopropylfluorophosphate (DFP). Substrate-affinity chromatography with phe-Sepharose or FMLP-Sepharose provided partial purification of enzyme activity among Mr 65,000 to 70,000 FMLP- or phe-binding proteins. Enzyme inhibition was obtained by incubation with mAb 1-15, DFP, N-carbobenzoxyl-phe-chlormethyl ketone, or PMSF, but not tosyl-amide-phenylethylchlormethyl ketone, bestatin, aprotinin, or phosphoramidon. In HPLC analysis, [3H]DFP labeled proteinase was found to comigrate with one of three FMLP-affinity-labeled membrane peaks, but unlike the FMLP surface receptor the DFP-labeling membrane proteinase was not modified by endoglycosidase F. We conclude that the mAb 1-15 Ag, which appears to play a role in PMN activation, is a distinct, active, Mr 65,000 to 70,000 serine proteinase with affinity for substrate sites containing aromatic amino acids.     

A parameter optimization approach for the optimal control of large-scale musculoskeletal systems.

This paper describes a computational method for solving optimal control problems involving large-scale, nonlinear, dynamical systems. Central to the approach is the idea that any optimal control problem can be converted into a standard nonlinear programming problem by parameterizing each control history using a set of nodal points, which then become the variables in the resulting parameter optimization problem. A key feature of the method is that it dispenses with the need to solve the two-point, boundary-value problem derived from the necessary conditions of optimal control theory. Gradient-based methods for solving such problems do not always converge due to computational errors introduced by the highly nonlinear characteristics of the costate variables. Instead, by converting the optimal control problem into a parameter optimization problem, any number of well-developed and proven nonlinear programming algorithms can be used to compute the near-optimal control trajectories. The utility of the parameter optimization approach for solving general optimal control problems for human movement is demonstrated by applying it to a detailed optimal control model for maximum-height human jumping. The validity of the near-optimal control solution is established by comparing it to a solution of the two-point, boundary-value problem derived on the basis of a bang-bang optimal control algorithm. Quantitative comparisons between model and experiment further show that the parameter optimization solution reproduces the major features of a maximum-height, countermovement jump (i.e., trajectories of body-segmental displacements, vertical and fore-aft ground reaction forces, displacement, velocity, and acceleration of the whole-body center of mass, pattern of lower-extremity muscular activity, jump height, and total ground contact time).