Chromosome analysis of BALB/c mouse spermatozoa with normal and abnormal head morphology.

To assess the relationship between mouse sperm head morphology and karyotype, sperm heads with either a normal or an abnormal morphology were injected individually into enucleated mouse oocytes that were karyotyped at the metaphase of the first cleavage. BALB/c male mice that produce an unusually high proportion of morphologically abnormal spermatozoa were used as sperm donors. Abnormal karyotypes were found in a significantly higher proportion of eggs injected with severely misshapen sperm heads (36-38%) as compared to those injected with normal and quasi-normal heads (15-21%) (p < 0.01). Most karyotype abnormalities were structural rather than numerical, the most common being breaks and exchanges of chromosome type in both normal and abnormal spermatozoa.     

The osmotic pressure of chondroitin sulphate solutions: experimental measurements and theoretical analysis

We used equilibrium dialysis to measure the osmotic pressure of chondroitin sulphate (CS) solutions as a function of their concentration and fixed charge density (FCD) and the ionic strength and composition of the solution. Osmotic pressure varied nonlinearly with the concentration of chondroitin sulphate and in 0.15 M NaCl at FCDs typical of uncompressed cartilage (approximately 0.4 mmol/g extrafibrillar H2O) was approximately 3 atmospheres. Osmotic pressure fell by 60% as solution ionic strength increased up to about 1 M, but remained relatively constant at higher ionic strengths. The ratio of Ca2+ to Na+ in the medium was a minor determinant of osmotic pressure. The data are compared with a theoretical model of the electrostatic contribution to osmotic pressure calculated from the Poisson-Boltzmann equation using a rod-in-cell model for CS. The effective radius of the polyelectrolyte rod is taken as a free parameter. The model qualitatively reproduces the non-linear concentration dependence, but underestimates the osmotic pressure by an amount that is independent of ionic strength. This difference, presumably arising from oncotic and entropic effects, is approximately 1/3 of the total osmotic pressure at physiological polymer concentrations and ionic strength.     

Proliferation of anchorage-dependent contact-inhibited cells. II: experimental results and validation of the theoretical models

This study establishes that the cellular automata models developed in an earlier article capture the essential features of the proliferation process for anchorage-dependent contact-inhibited cells. Model predictions are in excellent agreement with experimental data obtained with bovine pulmonary artery endothelial cells. The models are particularly suitable for predictive purposes since they have no adjustable parameters. All model parameters can be easily obtained from a priori measurements. Our studies also show that proliferation rates are very sensitive to the spatial distributions of seed cells. The adverse effects of seeding heterogeneities become more pronounced as a cell population approaches confluency and they should be accounted for in experimental studies attempting to assess the response of cells to external stimuli.     

Rotatory properties of molecules containing two peptide groups: experimental and theoretical results

Optical rotation studies were performed on five compounds containing two peptide groups. Four of the molecules were confined to restricted regions of conformational space by the presence of closed rings. Solvent and temperature were varied, and theoretical calculations were done for each compound covering the appropriate conformational space. The interpretation which results is qualitatively successful over the conformational regions covered by the compounds. Quantitative correlation between theory and experiment will require compounds of high rigidity.     

Direct chromosomal analysis of human spermatozoa: preliminary results from 18 normal men.

Chromosomal analysis of 240 spermatozoa from 18 normal men was performed using in vitro fertilization of zona-free golden hamster eggs. The frequency of chromosome abnormalities in this population was 9.2% (22/240). Of the abnormal complements, 18 were aneuploid (13 hyperploid and five hypoploid) and four had a chromosome break. The sex ratio of Y-bearing to X-bearing sperm was .68. The frequency and type of sperm chromosome abnormalities is compared with those seen in spontaneous abortions.     

Relationships between population size and loss of genetic diversity: comparisons of experimental results with theoretical predictions

Preservation of genetic diversity is of fundamental concern toconservation biology, as genetic diversity is required for evolutionarychange. Predictions of neutral theory are used to guide conservationactions, especially genetic management of captive populations ofendangered species. Loss of heterozygosity is predicted to be inverselyrelated to effective population size. However, there is controversy asto whether allozymes behave as predicted by this theory. Loss of geneticdiversity for seven allozyme loci, chromosome II inversions andmorphological mutations was investigated in 23 Drosophilamelanogaster populations, maintained at effective population sizesof 25 (8 replicates), 50 (6), 100 (4), 250 (3) and 500 (2) for 50generations. Allozyme genetic diversity (heterozygosity, percentpolymorphism and allelic diversity), inversions and morphologicalmutations were all lost at greater rates in smaller than largerpopulations. Conservation concerns about loss of genetic diversity insmall populations are clearly warranted. Across our populations, loss ofallozyme heterozygosity over generations 0–24, 0–49 and25–49 did not differ from the predictions of neutral theory. Thetrend in deviations was always in the direction expected withassociative overdominance. Our results support the use of neutral theoryto guide conservation actions, such as the genetic management ofendangered species in captivity.     

Graph-based analysis and visualization of experimental results with ONDEX

MOTIVATION: Assembling the relevant information needed to interpret the output from high-throughput, genome scale, experiments such as gene expression microarrays is challenging. Analysis reveals genes that show statistically significant changes in expression levels, but more information is needed to determine their biological relevance. The challenge is to bring these genes together with biological information distributed across hundreds of databases or buried in the scientific literature (millions of articles). Software tools are needed to automate this task which at present is labor-intensive and requires considerable informatics and biological expertise. RESULTS: This article describes ONDEX and how it can be applied to the task of interpreting gene expression results. ONDEX is a database system that combines the features of semantic database integration and text mining with methods for graph-based analysis. An overview of the ONDEX system is presented, concentrating on recently developed features for graph-based analysis and visualization. A case study is used to show how ONDEX can help to identify causal relationships between stress response genes and metabolic pathways from gene expression data. ONDEX also discovered functional annotations for most of the genes that emerged as significant in the microarray experiment, but were previously of unknown function.     

Binding of small basic peptides to membranes containing acidic lipids: theoretical models and experimental results.

We measured directly the binding of Lys3, Lys5, and Lys7 to vesicles containing acidic phospholipids. When the vesicles contain 33% acidic lipids and the aqueous solution contains 100 mM monovalent salt, the standard Gibbs free energy for the binding of these peptides is 3, 5, and 7 kcal/mol, respectively. The binding energies decrease as the mol% of acidic lipids in the membrane decreases and/or as the salt concentration increases. Several lines of evidence suggest that these hydrophilic peptides do not penetrate the polar headgroup region of the membrane and that the binding is mainly due to electrostatic interactions. To calculate the binding energies from classical electrostatics, we applied the nonlinear Poisson-Boltzmann equation to atomic models of the phospholipid bilayers and the basic peptides in aqueous solution. The electrostatic free energy of interaction, which arises from both a long-range coulombic attraction between the positively charged peptide and the negatively charged lipid bilayer, and a short-range Born or image charge repulsion, is a minimum when approximately 2.5 A (i.e., one layer of water) exists between the van der Waals surfaces of the peptide and the lipid bilayer. The calculated molar association constants, K, agree well with the measured values: K is typically about 10-fold smaller than the experimental value (i.e., a difference of about 1.5 kcal/mol in the free energy of binding). The predicted dependence of K (or the binding free energies) on the ionic strength of the solution, the mol% of acidic lipids in the membrane, and the number of basic residues in the peptide agree very well with the experimental measurements. These calculations are relevant to the membrane binding of a number of important proteins that contain clusters of basic residues.     

Heterosis and gene effects of multiplicative characters: theoretical relationships and experimental results from Vicia faba L.

Theoretical results were derived to relate the heterosis and the hybrid factor (ratio of hybrid performance to parental mean) of a complex character (seed yield) with the respective parameters of component subcharacters in a multiplicative model. A multiplication factor, which is a function of differences in the parents for subcharecters, was introduced to arrive at multiplicative relationships between the parameters in the model. Under certain assumptions, gene effects of a complex multiplicative trait can be expressed in terms of gene effects for the subcharacters. Data on seed yield and its components in two crosses between Vicia faba minor and major cultivars were used as a numerical example. Theoretical and experimental results indicate that with large complementary differences for subcharacters in the parents, it is possible to find substantial heterosis in the complex character without significant heterosis in its component traits. However, a review of results from the literature shows that multiplication effects are only of minor importance in most crops. Implications for the use of multiplication effects in the breeding of hybrid, synthetic, and line cultivars are discussed.     

Difference in sperm head volume as a theoretical basis for sorting X- and Y-bearing spermatozoa: potentials and limitations

Volume-based sorting of X- and Y-chromosome-bearing sperm cells could be an interesting alternative to the existing technique based on DNA content. Advantages would be that DNA staining and ultraviolet excitation, used in the existing technique, could be avoided. To assess the possibilities and limitations of sperm-head volume as sorting criterion, achievable purity and yield are determined for bull sperm. Two important parameters in this respect are the magnitude of the volume difference and the biological variation within each (X or Y) population. Earlier, we established a difference in volume matching the difference in DNA content (3.8%) between X- and Y-bearing bull sperm heads by comparing thicknesses and areas of high numbers of pre-sorted X- and Y-bearing bull sperm heads by interference microscopy and subsequent image analysis. Unfortunately, despite the high number of measurements, a direct determination of biological variations was not possible due to an unknown contribution of instrumental variations. In this paper, we determine the contribution of instrumental errors by measuring a single sperm head, varying parameters such as location in the image, orientation angle, focusing etc., simulating the behavior of the measuring system. After correction, both for the instrumental variation, and for the fact that the original samples were not pure, biological variations in volume of 5.9 +/- 0.8% were found. Our results indicate that when 10% of the bull sperm are sorted on basis of their head volume, a theoretical enrichment of 80% could be achieved. Expected purity and yield are lower than what is standard for the existing technique. At the moment, a technique to physically separate X- and Y-bearing sperm cells based on volume is not available. However, for applications for which the potential hazards of DNA staining and UV excitation are problematic, the development of such technique should be considered.     

Sensitivity analysis and optimization for a head movement model

A sixth order nonlinear model for horizontal head rotations in humans is analyzed using an extended parameter sensitivity analysis and a global optimization algorithm. The sensitivity analysis is used in both the direct sense, as a model fitting tool, and in the indirect sense, as a guide to experimental design. Resolution is defined in terms of the sensitivity table, and is used to interpret the sensitivity results. Using sensitivity analyses, the head and eye movement systems are compared and contrasted. Controller signal parameters are the most influential. Their variations and effects on head movement trajectories and accelerations are investigated, and the conclusions are compared with clinical neurological findings. The global optimization algorithm, in addition to automating the fitting of various types of data, is combined with time optimality theory to give theoretical time-optimal inputs to the model.On leave from Department of Neurology, University of Hamburg, FRG; supported by Deutsche Forschungsgemeinschaft Bonn, FRG     

Effects of distance constraints on macromolecular conformation. II. Simulation of experimental results and theoretical predictions

By generating classes of random structures for trypsin inhibitor and carp myogen, each consistent with a given set of experimental or theoretical information, we have assessed the relative utility of various experiments and theories in deducing the conformation of macromolecules. We compare the calculated structures with known x-ray coordinates and compute for each class an average error. Small errors mean that the experimental or theoretical constraints limit the structures to the vicinity of the crystal structure, whereas large errors show that the constraints permit a wide variety of tertiary conformations. We find the following points to hold true: (1) Qualitative information on all the distances, as might be obtained from the correct prediction of interresidue contacts, effectively determines the structure (error approximately 1 Å). (2) Quantitative information on a limited number of distances, as might be obtained from nmr or crosslinking experiments, significantly restricts the range of possible structures only when the number of distances given is comparable to the number of residues (error approximately 3 Å). (3) Quantitative information on the distances of each residue to the center of mass of the molecule, as might in part be obtained from solvent accessibility and solution x-ray studies, is not particularly restrictive by itself (error approximately 5 Å). (4) Complete qualitative local distance information, as might be obtained from secondary prediction and CD/ORD studies, is clearly consistent with a wide variety of tertiary structures (error approximately 7 Å).     

Chemiluminescent detection of bacteria: experimental and theoretical limits.

The limit of sensitivity of the chemiluminescent assay for detection of bacteria by hemeprotein catalysis of luminol oxidation was determined, both experimentally and theoretically, to be no lower than 10(5) to 10(6) viable bacterial per ml.     

Comparative analysis of movement characteristics of lancelet and fish spermatozoa having different morphologies

The movement characteristics of the sperm and their flagella obtained from a lancelet and 35 species from almost all orders of fishes were examined using high-speed video microscopy. The aim was to clarify the relationship between the motility parameters of the spermatozoa having different morphologies and how these motility parameters affect the swimming speed of the spermatozoa. The motility parameters representing the flagellar waveform, the wavelength, and the amplitude were neither very different between the spermatozoa of the different species nor related to the swimming speed. In contrast, the beat frequency was remarkably changed in the different spermatozoa and was proportional to the swimming speed. The maximum shear angle of the flagellar wave, which is directly related to the maximum sliding displacement between the doublet microtubules, remained nearly constant while the beat frequency varied widely; therefore, the spermatozoa beat in the constant sliding displacement mode. An analysis of the relationship between swimming speed and flagellar length revealed that short flagella were at a disadvantage in developing swimming speed; however, so were extra-long flagella. The ratio of the swimming speed to the wave velocity calculated from the wavelength and the beat frequency depended on the distance from the glass surface. The swimming speeds calculated using the original resistive-force theory were greater than the measured values. To rationalize the measured values, the ratio between the normal and tangential drag coefficient in the resistive-force theory was corrected; namely, 1.99 at 1 μm and 1.63 at 3 μm from the glass surface.     

Autocorrelation and spectrum of the modulated retinal discharge: A comparison of theoretical and experimental results

The theoretical autocorrelation of the cat ganglion cell discharge under stationary and dynamic conditions of light stimulus is compared with the autocorrelation determined experimentally. To obtain the theoretical autocorrelation, the stationary discharge is described by a stationary random point process of independent intervals equally distributed according to a gamma function, and the dynamic behaviour is described by a model defined in a previous paper. Comparison shows that the model predicts the experimental results. Finally, the power density spectrum is analysed and the relevance of the results to signal transmission by the retinal system is discussed.