Chicken major histocompatibility complex congenic lines differ in the percentages of lymphocytes bearing CD4 and CD8 antigens

In the experiments to be described two congenic inbred lines CB and CC and two recombinant lines CB.R1 and CC.R1 were used. All four lines differ only in regard to the major histocompatibility complex (MHC). To determine the percentage distributions of the two cell subsets in peripheral blood lymphocytes (PBL) in these lines, monoclonal antibodies to these two antigens were used. By FACScan there were more CD4+PBL in CB and CB.R1 lines (share B-F/B-L region, controlling class I/class II antigens with line CB) than CC and CC.R1, while the reverse was true with CD8+ subsets. There were more CD8+ PBL in the CC and CC.R1 lines and less in CB and CB.R1 lines. The ratio of CD4+ to CD8+ in CB chickens was 3.4 +/- 0.2 and in CC chickens 1.6 +/- 0.1.     

Long-term biological effects of air ions and D.C. electric fields on Namru mice: First year report

This report describes for the first time the effects of long-term continuous exposures of animals to small air ions and D.C. electric fields. In this study we exposed 200 female NAMRU mice (25/cage) to the following conditions: ± high ions (2×10⁵/cm³), ± low ions (2×10³/cm³), ± field only and ground (ion depleted, no field). Specially designed cages provided a defined D.C. field of about 2 kV/meter in ionized environments, with somewhat lower values in the field only cages. Detailed mapping of ion flux originating from a tritium foil generating system (multiple sources in an overhead plate) indicated a well defined, but heterogenous pattern with eight peak areas. Using a 100 cm² probe, ion flux values ranged from 10^–12–10^–14 A/cm², with an average flux of 8.7±6.8×10^–13 A/cm² in high negative ion cages, with good reproducibility between cages.Measurements of serum glucose, cholesterol, and urea nitrogen (samples taken every three months) showed a number of small but consistent and statistically significant differences between animals maintained in different environments during the first year of exposure. Serum globulin and whole blood serotonin, however, did not show any significant environmental effects. Interestingly, pairwise comparisons between high negative and low negative ion conditions, or between high positive and low positive ion conditions, or between the two ground conditions, revealed no significant differences between cages. This argues for a similarity of environmental responses for the mice maintained in each of the compared conditions.The results of a multiple classification analysis for the entire first year showed a preponderence of effects for the ionized cages, although other conditions also had highly significant differences as compared to the grand mean value. While this study has shown effects of only small magnitude (compared to normal physiological variations) in the female NAMRU mice studied here, the significance of these results strongly validates the precept of long-term air ion effects in animals. Given the known variability of different species and strains, future studies will have to determine the actual range of responsiveness of animals exposed on a long-term basis to defined air ion and electromagnetic environments.Deceased December 8, 1982.     

Spatially regulated translation in embryos: asymmetric expression of maternal Wnt-11 along the dorsal-ventral axis in Xenopus.

Transition from symmetry to asymmetry is a central theme in cell and developmental biology. In Xenopus embryos, dorsal-ventral asymmetry is initiated by a microtubule-dependent cytoplasmic rotation during the first cell cycle after fertilization. Here we show that the cytoplasmic rotation initiates differential cytoplasmic polyadenylation of maternal Xwnt-11 RNA, encoding a member of the Wnt family of cell-cell signaling factors. Translational regulation of Xwnt-11 mRNA along the dorsal-ventral axis results in asymmetric accumulation of Xwnt-11 protein. These results demonstrate spatially regulated translation of a maternal cell-signaling factor along the vertebrate dorsal-ventral axis and represent a novel mechanism for Wnt gene regulation. Spatial regulation of maternal RNA translation, which has been established in invertebrates, appears to be an evolutionarily conserved mechanism in the generation of intracellular asymmetry and the consequential formation of the multicellular body pattern.     

Purification and characterization of an ethanol-induced UDP-glucuronosyltransferase

A UDP-glucuronosyltransferase (GT) enzyme was isolated from ethanol-induced male New Zealand white rabbit hepatic protein. The animals were pretreated for 2 weeks with 10% ethanol in their drinking water. The GT enzyme was purified by anion-exchange and affinity chromatography and was shown to be homogeneous by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. The molecular mass of the ethanol-induced UDP-glucuronosyltransferase was determined to be 57,000 Da. Tryptic digests of the ethanol-induced GT and a similarly purified GT from control rabbit liver appeared to be different by HPLC analysis, even though the molecular masses of the enzymes were indistinguishable. Amino acid compositions of the two proteins were different for six amino acids. The apparent Km values for the ethanol-induced GT enzyme for 1-naphthol and morphine as substrates were 43 and 109 microM, respectively. The apparent Vmax values for the ethanol-induced GT enzyme for these substrates were 83 and 4.6 nmol/min/mg protein. The increases in catalytic efficiencies, apparent Vmax/Km for 1-naphthol and for morphine, for the ethanol-induced isozyme compared to the control isozyme activities were 2.0- and 2.4-fold. A polyclonal antibody raised in sheep to the rabbit ethanol-induced GT demonstrated a 520-fold selectivity for precipitation of the ethanol-induced protein rather than the control protein. These results demonstrate the production of an unique isozyme of UDP-glucuronosyltransferase that is produced in rabbits as a result of chronic ethanol exposure.     

Diaphragmatic thickness-lung volume relationship in vivo

To characterize the relationship of changes in diaphragmatic thickness during contraction to changes in lung volume, we developed a technique to measure diaphragm thickness based on M-mode ultrasonography. First, diaphragmatic thickness was measured in situ at necropsy with ultrasound and verified by measuring the same resected segment of diaphragm by ruler (correlation coefficient = 0.93, slope = 0.97). The technique of imaging the diaphragm in living subjects was developed by using a 15-MHz transducer coupled to an M-mode echocardiograph. Ten normal male volunteers were studied while sitting. The ultrasound transducer was held between the ribs in the ninth lateral interspace, and tidal volume was measured by spirometry. The thickening fraction (TF) was calculated as TF = (thickness at peak inspiration - thickness at end expiration)/thickness at end expiration for each of a series of different sized breaths. The function, TF vs. lung volume, for a range of volumes was linear for each subject and had intrasubject reproducibility with intersubject variability. We conclude that diaphragmatic TF is related to function as determined by lung volume, and this may prove to be a useful technique for in vivo studies of diaphragmatic function.