Analysis of haplotype preference in the cytotoxic T-cell response to H-Y

The mechanism determining which parental haplotype is selected in (CBA × 1310) (k × b)F₁ female mice for major histocompatibility complex (H-2) restricted, male-specific (H-Y), immune, cytotoxic T-cell (Tc-cell) responses, was investigated. The data show that haplotype preference is variable, and may be directed towards one, both, or neither of the parental haplotypes. This preference is reflected in the precursor frequency of memory Tc cells as measured by limiting dilution assays. It was further shown that maternal influence, antigen dose, route of immunization, and a feedback mechanism on the stimulator cells in vivo could not influence haplotype preference or its observed variability. Evidence for cross-reactive killing by H-2^k and H-2^b H-Y immune Tc cells on H-2^b and H-2^k allogeneic targets, respectively, (i. e., the independent haplotype of the other parent of the F₁ mice), provide evidence for natural tolerance as a possible mechanism to explain haplotype preference.     

A radioimmunoassay for 11-oxotestosterone: Its application in the measurement of levels in blood sebum of rainbow trout (S. Gairdneri).

17β-Hyd.roxyandrost-4-ene-3,11-dione was linked via its 3-(O-carboxymethyl) oxime to bovine serum albumin to give a conjugate which was used to generate antiserum in rabbits. The antiserum, at an overall dilution of 1 in 16,000, together with [1,2-³H] 17β-hydroxyandrost-4-ene-3,11-dione synthesized from [1,2-³H] cortisone have been used to develop a radioimmunoassay for the parent steroid. The assay incorporates a purification step in which serum or plasma extracts are chromatographed on silica gel layers bound to plastic or aluminium sheets and the steroid, containing zones cut out and eluted directly with assay buffer. The cross-reactivities of several steroids with the antiserum and the specificity, sensitivity, accuracy and precision of the assay are described. Blood sera from Immature male rainbow trout contain $\text{ca}$ 0.2–0.4 μg/100 ml of 17β-hydroxyandrost-4-ene-3,11-dione. As male fish mature, serum levels rise sharply to reach values of 2 to >9 μg/100 ml. Levels in immature females rarely exceeded the assay sensitivity but serum from three ripe females showed low but detectable levels ($\text{ca}$ 0.2 μg/100 ml) of steroid. The assay has found application in sexing live fish for experimental purposes.     

An animal model for the measurement of acute tolerance to ethanol

An apparatus is described for the measurement of acute tolerance to ethanol in small animals. Male, Sprague-Dawley rats were trained on the apparatus to leap to a descending platform to avoid being shocked. After an i.p. injection of 2 g/kg ethanol, the rats were tested repeatedly on the apparatus, and the plasma ethanol concentration was measured after each trial. The results demonstrated that the jumping ability of the rats was significantly more impaired during the ascending portion of the plasma ethanol curve than during the descending portion of the curve. Furthermore, it was demonstrated that the improvement in jumping ability during the descending portion of the curve was not dependent on a lowered plasma ethanol concentration. In a second experiment, the possibility of practice effects was eliminated by measuring the jumping ability and plasma ethanol concentration in one group of rats on the ascending portion of the plasma ethanol curve and in another group on the descending portion of the curve. A significant improvement in jumping ability was again observed during the descending portion of the curve, even though the plasma ethanol concentrations of the two groups were comparable. The development of acute tolerance to ethanol was thus demonstrated in both experiments.     

Hormonal control of deoxyribonucleic Acid and protein syntheses in pea root cortical explants

The hormonal control of DNA and protein syntheses in cortical explants taken at 10 to 11 mm from the tip of 3-day-old seedling roots (Pisum sativum cv. Little Marvel) was examined. On the auxin medium, S2M, the cortical cells began to enlarge at day 4 in culture, with no DNA synthesis or cell division throughout the 7-day culture period. With the addition of kinetin to this medium, S2M + K, the DNA content of the explants increased about three times by day 3, with further increases thereafter. This DNA increase was followed by cell division activity and subsequent tracheary element differentiation initiated at day 5. At least two divisions per parent cortical cell were required prior to this cytodifferentiation. The absolute hormonal requirements for the DNA synthesis and cell division responses were substantiated by the lack of either response in explants cultured on basal (S2M medium minus auxins) or basal + K medium for 7 days. On the auxin medium, there was no protein accumulation in the cortical explants over the 7-day period. On S2M + K medium, protein accumulation began after day 2 with a steady rate of increase until day 4, and some fluctuation thereafter. The pattern of increasing uptake of ¹⁴C-leucine was similar for days 0 to 4 in explants on either medium. After day 4 on S2M, the uptake continued to increase coincident with cell enlargement initiation, whereas on S2M + K there was a decline. Incorporation of ¹⁴C-leucine into trichloroacetic acid-precipitates of the total buffered homogenate from explants on both media exhibited a similar pattern, i.e. an increase during days 0 to 3 and then a decline to a level about three times higher than day 0. Incorporation into the homogenate soluble fraction also showed a similar pattern in explants cultured with or without kinetin. From the differences in net protein accumulation and the incorporation data, speculation on a cytokinin effect on protein synthesis and degradation rates is presented.     

The biology of the marine sponge Microciona prolifera (Ellis and Solander). III. Spicule secretion and the effect of temperature on spicule size

The secretion of siliceous spicules in the marine demosponge Microciona prolifera (Ellis and Solander) is by three different means. Styles are secreted by sclerocytes with archeocyte characteristics (nucleolate nucleus, phagosomes). chelas are formed by small sclerocytes with anucleolate nuclei, and toxas are apparently formed extracellularly within membranous material. Genetically and physiologically equivalent explants of this sponge were grown at 15, 20, and 25 C for four weeks. Analyses of spicule dimensions show little correlation of temperature with spicule length, except in the case of toxas. but a clear inverse relationship of spicule width with temperature. It is suggested that thicker spicules are formed at lower temperatures due to the more efficient entrapment of silicon rather than to effects upon silicon transport. Chela dimensions are very uniform implying an all or none process in their secretion. Differences in spicule dimensions between individual sponges grown at these temperatures may be due to the highly complex pathways of silicon transport and/or to genetic differences.     

Isolation and characterization of kinetoplast DNA and RNA of Phytomonas davidi

Kinetoplast DNA networks were isolated from stationary-phase culture forms of Phytomonas davidi. The networks banded in CsCl at a density of 1.699 g/ml and consisted of covalently closed circular molecules. The networks were sensitive to shear forces and exhibited several discrete sedimenting components in neutral and alkaline sucrose. Closed monomeric minicircles were isolated from sonicated networks by alkaline band sedimentation. Closed monomers showed a heterogeneous banding pattern on electrophoresis in acrylamide-agarose gels and had sedimentation coefficients of 20.5 S in alkaline sucrose and 11 S in neutral sucrose. The mean minicircle molecular weight as measured by cospreading with φXRF II was 0.70 × 10⁶ or 1064 nucleotide pairs. Minicircles exhibited a sequence microheterogeneity as evidenced by restriction enzyme analysis, melting analysis, and renaturation kinetics. Network maxicircles were evidenced by the appearance of high molecular weight fragments after restriction with several enzymes and by the existence of supertwisted “edge loops” extending out from the periphery of networks. The maxicircle molecular weight was estimated to be approximately 24 × 10⁶. A purified kinetoplast-mitochondrion fraction was found to contain 9 and 12 S RNA species that comigrated with L. tarentolae 9 and 12 S kinetoplast RNAs.     

The flight mechanism of the pigeon Columbia livia during take-off

High-speed film analysis of the mechanism of take-off of a pigeon ascending nearly vertically reveals the pattern of movements of the wing segments and the bones within them during each of the five phases of the wingbeat cycle. Differences in the type and extent of wing movements between the upstroke and downstroke portions of the first and successive wingbeat cycles are explained with reference to the upward vertical jump made during the first wingbeat cycle. The presence during pigeon take-off of a non-steady state pattern of airfoil action similar to that seen in some insects at the beginning of the downstroke was verified.