Protein diffusion in crowded electrolyte solutions

We report on a combined cold neutron backscattering and spin-echo study of the short-range and long-range nanosecond diffusion of the model globular protein bovine serum albumin (BSA) in aqueous solution as a function of protein concentration and NaCl salt concentration. Complementary small angle X-ray scattering data are used to obtain information on the correlations of the proteins in solution. Particular emphasis is put on the effect of crowding, i.e. conditions under which the proteins cannot be considered as objects independent of each other. We thus address the question at which concentration this crowding starts to influence the static and in particular also the dynamical behaviour. We also briefly discuss qualitatively which charge effects, i.e. effects due to the interplay of charged molecules in an electrolyte solution, may be anticipated. Both the issue of crowding as well as that of charge effects are particularly relevant for proteins and their function under physiological conditions, where the protein volume fraction can be up to approximately 40% and salt ions are ubiquitous. The interpretation of the data is put in the context of existing studies on related systems and of existing theoretical models.     

Testicular compensatory hypertrophy in the hemicastrated calf: effects of exogenous estradiol

Unilaterally orchidectomized (hemicastrated) bull calves were studied to monitor possible changes in serum concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) during the phase of testicular compensatory growth, to examine the characteristics of LH and FSH binding to the testis of the post-pubertal animal, and to determine whether any of these responses were altered by exogenous estradiol. Twenty-four calves were assigned randomly at one week of age to a 2 X 2 factorial experiment involving intact control (I) and hemicastrated animals (H), as well as estradiol-implanted intact (I+E2) and hemicastrated animals (H+E2). Relative to I, testis growth was accelerated in H and suppressed in I+E2 and H+E2. Mean testis weights at 27 weeks of age were 42 +/- 4, 72 +/- 6, 12 +/- 1 and 14 +/- 1 g for the four respective treatment groups. Serum FSH, but not serum LH, was positively associated with the accelerated testis growth of H. LH and FSH binding per testis were both enhanced approximately twofold in the testis from hemicastrated animals relative to those from intact calves. In contrast, estradiol markedly suppressed the number of LH-binding sites per testis in both I and H calves, but only suppressed the number of FSH-binding sites per testis in H calves. LH-affinity constants were not affected by treatment, whereas those for FSH were significantly decreased by estradiol. In conclusion, neonatal hemicastration results in elevated serum FSH, testicular compensatory hypertrophy, and an increased number of gonadotropin receptors in the bovine testis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Phenotypic analysis of thymocytes that express homing receptors for peripheral lymph nodes

Thymocytes that express high levels of homing receptors for peripheral lymph nodes can be detected with the monoclonal antibody MEL-14. We have shown that in adult mice these rare MEL-14hi thymocytes a) are cortical in location and typically constitute 1 to 3% of the total thymocyte population, b) may be a major source of thymus emigrants, and c) contain a high frequency of precursors of alloreactive cytotoxic T lymphocytes. In this study we have analyzed the phenotype of the MEL-14hi thymocyte subset. Most normal adult MEL-14hi thymocytes are midsize and express the mature phenotype typical of thymus emigrants, medullary thymocytes, and peripheral T cells: they are predominantly PNAlo, H-2K+, Thy-1+, Ly-1hi, and either Lyt-2-/L3T4+ or Lyt-2+/L3T4-. These findings argue strongly for the presence of rare MEL-14hi immunocompetent cortical thymocytes that, aside from their homing receptor expression, are phenotypically indistinguishable from medullary thymocytes. However, a minority (20 to 30%) of MEL-14hi thymocytes are large and phenotypically nonmature: they express intermediate to high levels of PNA binding sites, and are H-2K- to H-2Klo, Thy-1hi, Ly-1+, and either Lyt-2+/L3T4+ or Lyt-2-/L3T4-. Through a technique that selectively labels outer cortical cells, phenotypically nonmature MEL-14hi thymocytes have been shown to be concentrated in the subcapsular blast region of the outer cortex. Although we have no direct evidence of a precursor-product relationship, we consider it likely that the phenotypically nonmature outer cortical MEL-14hi lymphoblasts give rise to phenotypically mature MEL-14hi cells located deeper in the cortex. These results are consistent with our previous proposal that MEL-14hi thymocytes are a major source of thymus emigrants, and indicate that expression of high levels of MEL-14-defined homing receptors may be closely linked to the intrathymic selection process.     

In vivo visualization of individual neurons in arthropod ganglia facilitates intracellular neuropil recording

Summary A simple method for the in vivo visualization of dye filled cells by laser illumination is used to characterize neurons in situ in the segmentai ganglia of the locust and the crayfish (Fig. 1). Neuron visualization provides the structural information necessary for identification of cells during an ongoing physiological experiment (Figs. 2, 3). Sequential penetrations of soma and neuropil as well as simultaneous double neuropil penetrations of spiking and nonspiking cells are facilitated by the visual control afforded by neuron visualization (Figs. 4, 5, 6). Furthermore, neuron visualization allows the sampling of cellular properties at multiple, predetermined sites in the dendritic and axonal arbors of identified neurons (Fig. 7) and aids in establishing synaptic connectivity through double neuropil recordings (Fig. 8).