A polarized light and electron microscopic study of the birefringent fibrils inPhysarum plasmodia

Summary The birefringent fibrils in thin-spread plasmodium ofPhysarum polycephalum have been investigated with both polarizing and electron microscopes. The birefringent fibrils were classified into three groups by polarized light microscopy. The first type of fibril is observed in the advancing frontal region as a mutual orthogonal array. The birefringence changes rhythmically in accordance with the shuttle streaming. The second type of birefringent fibril is located in the strand region and runs parallel or somewhat oblique to the strand axis. The third type is observed in the strand region always perpendicular to the streaming axis. Electron microscopy confirmed that all these fibrils are composed of microfilaments, which range in densities in the cross view of the fibril from 1.2 to 1.7 × 10³/m² (1.5 × 10³/(xm² on the average).     

Modulation of glycosaminoglycan synthesis during cell growth as observed in an embryonic chick tendon cell culture

Glycosaminoglycan synthesis during cell growth has been studied in terms of unit cell numbers, using 16-day-old embryonic chick tendon cell cultures. Hyaluronic acid production was found to be inversely proportional to the cell density, while the levels of sulfated-glycosaminoglycan synthesis remained constant. On the other hand, hyaluronic acid production remained constant during cell proliferation, though chondroitin sulfate synthesis increased rapidly during an actively growing phase of the cultured cells, and dermatan sulfate and heparan sulfate syntheses increased gradually.     

Hydrolase production by Aspergillus niger in solid-state cultivation

Summary A production of macerating enzymes which liquefy and hydrolyze the mandarin orange peel was studied in a solid state cultivation of Aspergillus niger on wheat bran substrate. Solid state cultivation in a 2 drum fermenter capable of interchangeable operation under dynamic or static conditions were carried out maintaining the moisture content of the substrate at 32, 39, 46, 56, 67, and 74%. Biomass grown on the solid substrate was estimated on the basis of a constant value of glucosamine content of A. niger, 50 mg glucosamine/g cell. A linear relationship between oxygen uptake rate and growth rate observed in all the experiments gave an oxygen growth yield, Y_X/O, of 28.5 g cell/mol O₂. The rate of macerating enzyme formation was also in proportion to the growth rate irrespective of the difference of the moisture content of the substrate.The enzyme accumulation on the solid substrate, the growth rate and oxygen uptake rate were maximum when the moisture content of the substrate was maintained at ca. 56% ascending from 32 to 56 and descending from 56 to 74.     

Structural specificity of synthetic peptide adjuvant for induction of experimental allergic encephalomyelitis

The peptide N-acetylmuramyl-l-alanyl-d-isoglutamine (MDP), which has adjuvant activities, and 17 of its derivatives and analogs were synthesized and assayed to elucidate the structure necessary for adjuvant activity in induction of experimental allergic encephalomyelitis (EAE) in guinea pigs. The results revealed the importance of the d configuration and the α-carboxamide group of the isoglutaminyl residue of MDP for adjuvant activity. Replacement of the l-alanyl residue of MDP by d-alanine, but not by l-serine or glycine, resulted in a marked decrease in the activity. The β-methyl glycoside of MDP was found to be more active than the α-methyl derivative. 6-O-Stearoyl-N-acetylmuramyl-l-alanyl-d-isoglutamme showed activity.     

Counteracting effect of eosin and related dyestuffs on the production of respiration-deficient mutants in yeast by 4-nitroquinoline 1-oxide

Production of respiration-deficient (rho^?) mutants under growing conditions in a strain of Saccharomyces chevalieri by 4-nitroquinoline 1-oxide (4NQO), a potent carcinogen, reached 100%. the mutation frequency was considerably reduced when eosin Y was applied in various combinations with 4NQO. The counteracting effect was slight when eosin Y was applied concurrently with 4NQO, but was very strong adn persistent when eosin Y was impregnated into the yeast cells before their exposure to 4NQO. Eosin B, erythrosin B and uranin also showed more or less counteracting effects against 4 NQO in producing the rho^? mutants. Possible mechanisms for the counteracting effects of these dyestuffs against 4NQO are discussed in relation to antimutagenesis and chemotherapeutic interference.     

Hereditary respiration deficiency in Saccharomycodes ludwigii

Saccharomycodes ludwigii, supposed to be petite-negative, gave rise to respiration-deficient mutants when acriflavine and ultraviolet irradiation, respectively, were applied to this yeast, strain IFO 1194. The frequency of such mutants was very low as compared with that in Saccharomyces cerevisiae and other petite-positive yeasts. Cytochrome composition was characterized by spectrophotometry at the temperature of liquid nitrogen. The respiratory mutants examined contained cytochrome c unaltered in quality and quantity. Cytochrome b was often present only in small amounts though never absent, while cytochrome a+a₃ was either present or absent. The respiratory mutants could form zygotes after conjugation with a wild-type culture of opposite mating type ( vs. a). The hybridization and segregation analysis of spore tetrads showed the inheritance of respiratory mutant character to be either Mendelian or non-Mendelian and similar to that of pet (nuclear) and rho^- (cytoplasmic) mutants, respectively, in Saccharomyces cerevisiae.     

Genetic analysis of nurse dams selected for six-week body weight or postweaning gain in mice.

A modified crossfostering technique was developed to compare the performance of nurse dams in selected and control populations of mice. The H6 and M16 populations were selected for increased 6-week body weight and 3- to 6-week postweaning gain, respectively, while the C2 and ICR populations were the respective controls. Crossfostering was performed using H6, M16 and their reciprocal F1 crosses as nurse dams in the selected crossfostering group and C2 ICR and their reciprocals in the control group. Measurements recorded for nurse dams included mean body weight of 8 young within a nursed litter at birth (MWB) and 12 days of age (MW12). The latter was used as a measure of postnatal maternal performance. Other traits recorded for nurse dams were number born (NB), body weight at parturition (DWP) and 12 days postpartum (DW12), and weight gain (DWG), feed intake (FED) and efficiency (EFF = DWG/FED) for the first 12 days of lactation. The correlated response in MW12 was negative (P less than .01) for M16 and essentially zero for H6. Both lines exhibited positive (P less than .01) correlated responses in DWP and DW12 and no change in EFF. Only the H6 line increases significantly in DWG and FED as a result of selection. NB increased in M16 and H6, but was significant for the latter population only. Population differences in selection response [(M16-ICR)-(H6-C2)] were significant for FED only, primarily due to average direct genetic effects. Direct comparisons of M16 and H6 indicated that M16 was larger in DWP and DW12 but smaller in DWG and EFF. Average direct genetic effects favored M16 for NB, DWP, and DW12, whereas average maternal genetic effects favored H6 for NB, DWP, DW12 and FED. Percent direct heterosis, in F1 crosses of selected populations was significant for MW12 (13.7%) ,FED (10.8%) and NB (11.4%). Direct heterosis in F1 crosses of the controls was significant for MW12 (9.4%), NB (6.6%), DWP (3.5%), DW12 (3.3%) and FED (4.4%). The effects of MW12, DWG and metabolic body size (MBS) accounted for 47% of the variation in FED, pooled within populations. Of these variables, MW12 accounted for the highest proportion (32%) of variation in total feed intake.     

Studies on the circadian rhythm of phosphoenolpyruvate carboxykinase. III. Circadian rhythm in the kidney.

Phosphoenolypyruvate carboxykinase [EC 4.1.1.21] activity in rat kidney shows a circadian rhythm with the highest activity between 0200 h and 0800 h and the lowest activity between 1400 h and 2000 h. The rhythm was observed in both sexes and throughout the year. Actinomycin D and cycloheximide effectively blocked the circadian increase in enzyme activity. These findings suggest that the circadian increase in phosphoenolypyruvate carboxykinase activity is due to net synthesis of enzyme protein through newly synthesized mRNA. In experiments with kidney cortex slices, gluconeogenesis from the radioactive precursor, [14C]malic acid, was considerably higher at 0200 h than at 1400 h, varying in parallel with the change in the enzyme activity.     

Phosphorylation of mononucleotides and formation of cytidine 5''-diphosphate-choline and sugar nucleotides by respiration-deficient mutants of yeasts.

Respiration-deficient mutants (Rho-, petite) of Saccharomyces carlsbergensis were obtained by treatment with trypaflavin (euflavine). Dried cells of these mutants phosphorylated mononucleotides to their triphosphates and further formed not only cytidine 5'-diphosphate-choline, but also sugar nucleotides, such as uridine 5'-diphosphate-glucose, guanosine 5'-diphosphate-mannose, etc. The activities were the same or slightly greater than those of the wild strain. These results showed that energy (adenosine 5'-triphosphate) necessary for phosphorylation of mononucleotides was sufficiently supplied by the glycolysis system.