Muramic Acid Assay in Sediments

An improved chromatographic assay for muramic acid which is sufficiently sensitive for marine sandy sediments is described; it involves acid hydrolysis, thin-layer chromatography, and gas-liquid chromatography.     

Reduced color polymorphism in a population of Bunodosoma granulifera

The proportions of carotenoids responsible for column coloration were determined for a population of the sea anemone Bunodosoma granulifera from Grand Cayman using TLC. This population exhibited a very low degree of polymorphism for these proportions, compared to previously sampled populations from Puerto Rico and Curaçao. In addition to the carotenoid pigment system a second system possessing pigments similar to bilins was found for the first time in this genus. This population was also assayed electrophoretically for enzymes encoded by 12 loci. Analysis of these data suggested that the reduced polymorphism for column color may be the result of selection. Although the proportions of carotenoids varied little, the amount of carotenoid per unit of epidermal tissue varied almost four-fold from 0.05 to 0.19 μg/mg dry wt.     

The effect of propylthiouracil (PTU) on heat production in chickens maintained on different levels of nutrition

Using the method of forced feeding on higher and lower levels, two groups of chickens differring in growth rate were obtained. In the chickens of the two groups 72 measurements of heat production were made. Chickens of these groups were compared considering either the same age or similar body weight. It has been found that propythiouracil treatment (120 mg/kg duging 10 consecutive days) decreases heat production in the chickens of both groups. However, this effect was much more pronounced in the birds maintained on the higher level of nutrition and growing more rapidly than in those maintained on the lower level of nutrition and growing slowly.     

Photochemical oxidation of tyrosinase

Dye sensitized photo-oxidation inactivates tyrosinases isolated from Neurospora and Agaricus. The rate of inactivation is enhanced by cyanide and is dependent on pH.     

Studies of the biosynthesis of actinomycin in protoplasts from Streptomyces antibioticus.

Protoplasts actively synthesizing actinomycins have been prepared from Streptomyces, antibioticus. They showed an absolute requirement for the presence of oxygen, galactose, and alkaline earth ions. Sucrose was most efficient as an osmotic stabilizer. However, in air-saturated buffer the protoplasts seemed to be slightly inhibited in their metabolism. This is expressed by the appearance of 4-methyl-3-hydroxyanthranilic acid and the inability to utilize [1?¹⁴C]sarcosine for actinomycin synthesis. Evidence has been obtained that sarcosine and N-methyl-l-valine are not free precursors of the peptide-bound N-methyl-amino acids. It is further demonstrated that synthesis of actinomycin IV and actinomycin V differ from each other with respect to their different susceptibilities against the changings in the physiological environment of the protoplasts. Actinomycin synthesis is severely reduced when protoplasts are incubated in the presence of 10^?3, m methionine.     

Neural crest cell behavior in white and dark embryos of Ambystoma mexicanum: Epidermal inhibition of pigment cell migration in the white axolotl

Melanophores in larvae of the white (dd) strain of the Mexican axolotl (Ambystoma mexicanum) are confined to the dorsal midline of the trunk and dorsal posterior part of the head, whereas those in dark larvae (D_-) are distributed over the flank as well. Our results show that this phenotype of white larvae is the result of the failure of the melanophores or their neural crest precursor cells to migrate laterally due to an inhibition of or a failure in the support of their migration in the subepidermal space by the overlying epidermis. Correlated light and scanning electron microscopy of dissected larvae showed melanophores occupying the subepidermal space on the flank of dark larvae, whereas these cells were restricted to the dorsal midline of white larvae. Grafting experiments in which patches of epidermis, the underlying mesoderm, or both, were exchanged between dark and white embryos suggested that white epidermis alone can prevent the integration of pigment cells on the flank of dark larvae and, conversely, that grafts of dark epidermis alone can support their migration on the flank of white larvae. Mesoderm, when grafted alone, could not be shown to have similar effects.     

Characterisation and Regional Localisation of Pre- and Postsynaptic Glycoproteins of the Chick Forebrain Showing Changed Fucose Incorporation Following Passive Avoidance Training

To identify those glycoproteins whose synthesis or modification is necessary for memory formation, we have studied the uptake of radiolabelled fucose into synaptic plasma membranes (SPMs) and postsynaptic densities (PSDs) derived from two specific left and right forebrain loci, at two different times after training of 1-day-old chicks on a one-trial passive avoidance learning task. To increase the reliability of the comparison, a double-labelling method was used. Tissue samples from intermediate medial hyperstriatum ventrale (IMHV) and lobus parolfactorius (LPO) were isolated at 6 and 24 h after training. At both times, training resulted in region-specific changes, both increases and decreases, in incorporated radioactivity into pre- and postsynaptic glycoproteins. After 6 h, there was a relative decline in incorporation into both SPMs and PSDs of the right IMHV of trained chicks, a decline that persisted in the PSDs until 24 h. A small decline in incorporation in SPMs from the right LPO of trained chicks at 6 h was reversed by 24 h, by which time there was a 64% increase in incorporation into SPMs and a 24% increase into PSDs of the left LPO. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis of left and right hemisphere samples containing LPO revealed that 6 h after training the main effect was presynaptic, including a reduction of incorporation into high molecular mass glycoproteins, of 150-180 kDa, and an increase in a lower molecular mass (41 kDa) fraction. By 24 h after training, a left hemisphere presynaptic glycoprotein of molecular mass approximately 50 kDa showed the biggest increase in fucosylation. In addition, a wide group of postsynaptic glycoproteins of both hemispheres, in the ranges 150-180, 100-120, and 33 kDa now showed increases in incorporation. Some other fractions showed decreases. These results are in accord with previous data on incorporation obtained using the amnesic agent 2-deoxygalactose. They also support the hypothesis that memory formation involves the strengthening of connections between pre- and postsynaptic neurons of the LPO by growth or modulation of pre- and postsynaptic structures.     

Amino acid substitutions at tryptophan 388 and tryptophan 412 of the HepG2 (Glut1) glucose transporter inhibit transport activity and targeting to the plasma membrane in Xenopus oocytes.

All 6 tryptophan residues in the human HepG2-type glucose transporter (Glut1) were individually altered by site-directed mutagenesis to investigate the role of these residues in transport function. Tryptophan residues in positions 48, 65, 186, 363, 388, and 412 of Glut1 were changed to either a glycine or leucine residue. Mutant mRNAs were synthesized and injected into Xenopus laevis oocytes. Transporter function as assessed by uptake of 2-deoxy-D-[3H]glucose or transport of 3-O-[3H]methylglucose was decreased in the 388 and 412 mutants but was unaltered in all other mutants. The amount of the mutant transporters expressed in total membrane and plasma membrane fractions was measured using Glut1-specific antibodies. Calculation of the intrinsic transport activity of each of the mutants using these data demonstrated that the reduced transport activity of the 412 mutants was caused entirely by a dramatic decrease in the intrinsic activity of the mutant proteins whereas the reduced activity of the 388 mutants was a result of a decreased level of the protein in oocytes, decreased targeting to the plasma membrane, and a modest decrease in the intrinsic activity. Protease/glycosidase mapping of in vitro translation products indicated that the effects of the 388 and 412 point mutations could not be attributed to a disruption in the ability of the mutant proteins to insert properly into the membrane. The ID50 for cytochalasin B inhibition of 2-deoxyglucose uptake was increased from 5 x 10(-7) M for the wild-type Glut1 to 4 x 10(-6) M in the 388 mutants but was unaltered in the 412 mutants. These observations suggest that 1) Trp-412 may comprise part of a hexose binding site or is involved in maintaining a local tertiary structure critical for transport function; 2) Trp-388 is involved in stabilizing the equilibrium binding of cytochalasin B to the transporter. Trp-388 may therefore lie near a substrate binding site and also appears to participate in stabilization of local tertiary structure important for full catalytic activity and efficient targeting to the Xenopus plasma membrane.