Rapid,gregarious settlement of the larvae of the marine echiuran Urechis caupo Fisher & MacGinitie 1928

Larvae of Urechis caupo Fisher & MacGinitie, reared in the laboratory, were exposed to potential settlement stimuli, including natural sediment from adult burrows, and “scent” obtained from the skin of adult animals. Competent larvae settled rapidly and specifically in response to adult burrow sediment when compared with their responses to other natural and abiotic sediments. Larvae also responded specifically to chemical “scent” from adult animals when the “scent” of another echiuran worm, Listriolobus pelodes Fisher served as a control. Larval responses to chemical “scent” were as great as their responses to natural burrow sediment. Hence, it is likely that larvae settle gregariously in nature in response to “scent” on sediment grains of adult burrows. The chemical “scent” had a molecular weight between ≈3500 and 14000 daltons, as determined by dialysis. It quickly lost its effectiveness in promoting settlement after it was heated to 80 °C, but was relatively stable at ambient ocean temperatures, retaining its effectiveness for several days. It was soluble in sea water. However, larvae did not respond to the chemical “scent”, unless it was adsorbed onto a surface. Purely tactile stimuli, such as the shape, texture, and size-distribution of particles, were not important settlement cues during these experiments.     

Characterization of early B lymphocyte precursors present in long-term bone marrow cultures

Lymphoid precursor cells are present in long-term bone marrow cultures (LTBMC), but their differentiation into mature lymphocytes is blocked. A quantitative assay for B cell precursors in LTBMC, which gives a linear relationship between the number of grafted LTBMC cells and the frequency of B cell colony forming units (CFU-B) in the spleen and bone marrow of immunodeficient CBA/N mice 19 days after reconstitution, is described. Characterization of the B cell precursor indicates that this assay is detecting a very early precursor and not a B lymphocyte or a late pre-B cell. This conclusion is based on the observations that a) pre-B cells transformable by Abelson murine leukemia virus are not present in LTBMC by 3 days postrecharge and CFU-B are absent by 6 days postrecharge; b) late B cell progenitors capable of rapid repopulation of irradiated CBA/N mice are not present in LTBMC, since a lag in the kinetics of B cell reconstitution in animals grafted with LTBMC cells is observed compared with fresh bone marrow cells; c) the B cell precursors in LTBMC have high proliferative potential, since they can stably repopulate recipient mice for at least 8 wk postreconstitution and through two serial passages in irradiated CBA/N recipients; and d) the B cell precursors are large, rapidly sedimenting cells as determined by velocity sedimentation. The serial transplantation experiment further shows that a split is often observed between lymphoid and myeloid reconstituting ability of LTBMC cells. The LTBMC B cell precursor may be a pluripotent stem cell or a lymphoid stem cell, although its differentiative potential remains to be determined.     

Lymphocyte zinc metabolism in disease

Lymphocytes were obtained from two patients with paroxysmal nocturnal hemoglobinuria as well as from apparently healthy controls and from patients with acute lymphoblastic leukemia and chronic lymphocytic leukemia. Subsequently, several aspects of zinc metabolism were studied in vitro in short-term cultures of these lymphocytes in order to assess lymphocyte functional capacity. The results of mitogen stimulation and zinc uptake studies for lymphocytes from donors with paroxysmal nocturnal hemoglobinuria were similar to those obtained for leukemic lymphocytes. The results of studies to determine the inducibility of the low molecular weight zinc-binding protein, metallothionein, by zinc were complicated by the decrease in overall protein synthesis in lymphocytes from donors in the paroxysmal nocturnal hemoglobinuria. It is proposed that paroxysmal nocturnal hemoglobinuria is indeed a clonal disorder and the relationship between lymphocytes in this disorder and leukemic lymphocytes is discussed.     

Crystallisation and preliminary X-ray data of ribulose-1,5-bisphosphate carboxylase from spinach

Crystals of a tertiary complex of spinach ribulose-1,5-bisphosphate carboxylase/oxygenase with the activators Mg²⁺ and CO₂ have been grown. These crystals diffract strongly to 1.6 Å resolution. The spacegroup is C222₁ with unit cell dimensions a = 158.6 Å, b = 158.6 Å, c = 203.4 Å. Additional local symmetry is apparent in the pattern of absences and the intensity distribution of the X-ray precession photographs. The photographs have been interpreted in terms of a molecule (consisting of eight large and eight small subunits, L₈S₈) with 222 symmetry and a molecular centre shifted 2 Å in the x direction from the origin of the unit cell. The asymmetric unit contains half the L₈S₈ molecule. The intensity distribution suggests that the molecular symmetry does not deviate far from 422. These crystals are compared with other crystalline forms of the enzyme and the implications of these results are discussed.     

Sexual differentiation of rat liver carbonic anhydrase III

Using radioimmunoassay, the concentration of carbonic anhydrase III in the livers of adult male rats was found to be approx. 30-times greater than that observed in mature females. Castration of male rats led to a marked reduction in liver carbonic anhydrase III concentrations which could be partially restored to control levels by testosterone replacement. Administration of testosterone to ovariectomised female rats induced about a 5-fold increase in liver carbonic anhydrase III concentration. Immunoprecipitation analysis of the products of liver mRNA translation in vitro with antiserum specific for carbonic anhydrase III showed that hormonal control of the levels of carbonic anhydrase III in liver is mediated by changes in the amount of translatable carbonic anhydrase III mRNA. Marked changes in liver carbonic anhydrase III concentrations were also observed in developing and ageing male rats.     

Association of newly synthesized major fl coat protein with infected host cell inner membrane

The bacteriophage fl major coat protein becomes associated with the host cell inner membrane very shortly after it is synthesized. Pulse-chase experiments suggest that the virus is never stably associated with the host cell outer membrane; we propose that it passes directly from the inner membrane to the growth medium.     

Involvement of Threonine Dehydratase in Biosynthesis of the α-Ketobutyrate Prosthetic Group of Urocanase

Seventeen mutants of Pseudomonas putida that were unable to grow on threonine as nitrogen source owing to a lack of threonine dehydratase were isolated, and all were found to be unable to synthesize active urocanase. Spontaneous revertants selected for urocanase production concomitantly regained threonine dehydratase. Mutants that were unable to utilize urocanate as carbon source were also isolated, and these were defective in urocanase formation but were normal in threonine dehydratase levels. Since alpha-ketobutyrate is the prosthetic group for urocanase, these results are consistent with the proposal that threonine dehydratase is necessary for urocanase prosthetic group biosynthesis. However, the lack of urocanase activity in threonine dehydratase-negative mutants was shown not to be the result of reduced levels of endogenous free alpha-ketobutyrate, nor to the participation of threonine dehydratase in the initiation of urocanase biosynthesis through the conversion of threonyl-tRNA(Thr) to alpha-ketobutyryl-tRNA(Thr). Other alternatives for the participation of threonine dehydratase in urocanase biosynthesis are discussed.     

Metabolism of n-Butane and 2-Butanone by Mycobacterium vaccae

n-Butane was metabolized in Mycobacterium vaccae (JOB5) via terminal oxidation. This organism metabolized 2-butanone through propionate (or propionyl coenzyme A). Subterminal oxidation in M. vaccae was apparently limited to propane.     

Steady-state kinetics of catecholamine transport by chromaffin-granule `ghosts''

Resealed chromaffin-granule ;ghosts' were used to study the steady-state kinetics of catecholamine transport. The pump has a high affinity for (-)-noradrenaline, (-)-adrenaline, tyramine and 5-hydroxytryptamine (serotonin), but a lower affinity for (+)-noradrenaline. The measured rates of incorporation do not conform to Michaelis-Menten kinetics, but affinity constants for the former substrates are in the range 8-18mum. Reserpine is a potent inhibitor. Incorporation as a function of ATP concentration also fails to show simple kinetics; the affinity constant for ATP is deduced to be about 3mm at 1mm-MgCl(2). Adenylyl (betagamma-methylene)diphosphonate is a competitive inhibitor at low concentrations, but inhibits more strongly at high concentrations. The pump has a transition temperature at 29 degrees C and does not seem to be identical with the Mg(2+)-stimulated adenosine triphosphatase of chromaffin granules.     

Transport of catecholamines by resealed chromaffin-granule `ghosts''

A method is described for the preparation of resealed chromaffin-granule ;ghosts'. The lysis and rapid purification procedures provide ;ghosts' in approximately 70% yield from crude granules; the preparation contains 0.1mumol of catecholamine/mg of protein (as compared with 2.8mumol/mg in unlysed granules), of which about one third is inside the ;ghosts'. The ;ghosts' retain their ability to accumulate catecholamines, a process dependent on Mg-ATP and inhibited by reserpine, and a simple assay for this transport is described.