Changes in Phospholipid Composition of a Winter Wheat Cultivar during Germination at 2 C and 24 C

Evaluation of various solvent systems for lipid extraction of wheat Triticum aestivum L. cv. Rideau seeds showed that boiling 2-propanol followed by the Bligh-Dyer procedure was the most efficient method, with respect to lipid yield and ability to inactivate lipolytic enzymes. Ten phospholipids were identified in dry seeds; the major components being phosphatidylcholine, lysophosphatidylcholine, N-acyl lysophosphatidyl-ethanolamine, N-acylphosphatidylethanolamine, and phosphatidylethanolamine. After growth for 1 week (2 C) or 31 hours (24 C), the proportions of phosphatidylethanolamine + lysophosphatidic acid and phosphatidic acid increased, lysophosphatidylcholine decreased, and the remaining phospholipids showed little change. At 5 weeks (2 C) or 72 hours (24 C), the seedlings showed 5-fold increases in the proportion of phosphatidic acid largely at the expense of phosphatidylcholine, small decreases in N-acyl lysophosphatidylethanolamine and N-acylphosphatidylethanolamine, and significant increases in lysophosphatidylcholine. The changes in phosphatidic acid and phosphatidylcholine are interpreted as being partially due to increasing phospholipase D activity during germination. In general, the phospholipid composition was similar in morphologically equivalent seedlings grown at 2 C or 24 C. The increased membrane content in seedlings grown at 2 C does not reflect any preferential synthesis of individual phospholipids.     

Relative insensitivity of mitochondria in hardened and nonhardened rye coleoptile cells to freezing in situ

Mitochondria were isolated from excised coleoptiles of hardened and nonhardened winter rye (Secale cereale L. cv. Puma) seedlings which had been frozen extracellularly to different temperatures. No significant differences in the respiratory functions (ADP/O and respiratory control) were observed between mitochondria isolated from nonlethally and lethally frozen cells of both the hardened and nonhardened rye. These results suggest that mitochondria in situ can retain their normal function even after the cell was killed by the dehydrative stresses of extracellular freezing. Presumably, a different level of sensitivity to freezing stresses exists between mitochondrial and other membranous elements in the cell.     

Differential scanning calorimeter analyses of membrane lipids isolated from hardened and unhardened black locust bark and from winter rye seedlings

Phospholipids isolated from hardened and unhardened cells of the living bark of the black locust tree and from the cells of winter rye seedlings were analyzed by differential scanning calorimetry. In accord with previously reported data which showed little differences in the fatty acid unsaturation of phospholipids obtained from unhardy summer and very hardy winter bark of the black locust, only small differences were found in the temperature and enthalpies of the liquid-crystalline gel phase transitions of the isolated phospholipids of these respective tissues. In the case of winter rye, where fatty acid unsaturation was observed to increase with hardening, the differences in phase transition properties of the phospholipids isolated from hardened and unhardy rye also were found to be minor. In any event, the transition temperatures of the rye phospholipids were well above and that of the locust phospholipids far above the freeze-killing temperatures for these tissues. These results would suggest that both unsaturation and bulk fluidity of the isolated lipids from these plants bear little relationship to their degree of freezing tolerance.     

The determination of the binding constant of metalloenzymes for their active site metal ion from ligand inhibition data: Theoretical analysis and application to the inhibition of thermolysin by 1,10-phenanthroline

An equation is found relating the fractional activity, (v/v₀), of an enzyme assay mixture to the total concentrations of metalloenzyme, active site metal ion, metal-binding ligand and substrate and the stability constants of the complexes present. When (v/v₀) is measured as a function of the total ligand concentration, this equation offers a way of data-plotting which yields straight lines and permits the calculation of the metal-binding constant K_ME from either the slope or the intercept, provided that mixed complexes (enzyme-metal ion-ligand) do not contribute significantly to the change in (v/v₀). Since deviations from linearity occur in the latter case, the proposed inhibition plot serves as a diagnostic tool for the recognition of such complexes. Application to the inhibition of thermolysin by 1,10-phenanthroline gives a value of 2.1 × 10¹¹m⁻¹ for K_ZnE, the binding constant of the active site zinc ion, at pH 7.50, 25°C and ionic strength 0.1. The equation also allows the rapid calculation of the ligand concentration necessary to attain a desired degree of inhibition when the total enzyme and active site metal ion concentrations of the solution are known.     

Associations between the rates of maturation of the bones of the hand-wrist

Skeletal maturation rates for the age interval 3 to 13 years were analyzed using bone-specific assessments (Greulich-Pyle) of serial radiographs of 40 children. The mean rates of skeletal maturation resembled those of the population from which the atlas standards had been derived. There was a linear trend of skeletal age against chronological age for most bones in each sex. Regression lines were fitted to these data and the b values of the regression lines were calculated. Communality indices were calculated from an intercorrelation matrix of these b values. There was a statistically significant rank order correlation between the sexes in the communality indices. They tended to be higher in the girls than in the boys and were relatively low for the radius, ulna and carpals. Communality indices within groups of bones were high in all rows, especially the metacarpals, but in each sex they were comparatively low in the first ray (metacarpal plus the phalanges of the corresponding digit) and in the fifth ray of the boys. Neighborhood effects on the levels of association of maturation rates were present, particularly in the carpus, but marginal effects were not noted.     

Correlations between rates of epiphyseal and diaphyseal elongation in the short bones of the hand

Serial annual radiographs of the hand have been used to analyze the rates of elongation of the epiphyses and diaphyses of the metacarpals and phalanges in children at ages from 3 to 13 years. The rates of elongation for many corresponding ephiphyses and diaphyses (i.e., of the same bone) are negatively correlated but to an extent that is not statistically significant for any particular bone. This tendency toward negative correlations is found for most of these bones although the correlation coefficients for most of the metacarpals are positive in each sex. Within rays, the correlation indices between the rates of elongation for corresponding epiphyses and diaphyses (i.e., of the same bone) have larger negative or smaller positive values than for those between either adjacent and non-corresponding or non-adjacent and non-corresponding epiphyses and diaphyses (i.e., not of the same bone but of either adjacent or non-adjacent bones). The communality indices for the ratio between the rates of epiphyseal and diaphyseal elongation in particular bones are more highly correlated in the girls than in the boys and within the rows than within the rays. Some implications of the tendency to negative correlations between the rates of elongation of corresponding epiphyses and diaphyses have been discussed.     

Isolation and study of a strain of Candida tropicalis growing on n-alkanes

Summary A strain of Candida tropicalis has been isolated from soil using a mineral medium that contained n-tetradecane as sole source of carbon. This strain has been studied and variants have been isolated. In contrast to the original strain in which hydrocarbon degradation is linked to enzymatic induction mechanisms, the variant 101 behaves like a constitutive strain for n-tetradecane.     

Effect of progesterone on basal LH and episodic LH and FSH secretion in heifers

Heifers between Days 6 and 10 of the cycle were allocated at random to groups of 8 and treated with (i) a 4% progesterone-releasing intravaginal device (PRID) + oestrogen capsule for 12 days; (ii) 4% PRID for 12 days; (iii) 20% PRID for 12 days; (iv) 4% for 14 days; or (v) 20% PRID for 14 days. Blood was obtained daily during treatment and at 2- or 4-h intervals for 72 h after removal of PRIDs. Some animals were sampled every 20 min for 4.676 h on the 3rd day after PRID insertion, and 1 day before and 36 h after removal of the PRID insertion, and 1 day before and 36 h after removal of the PRID. During progesterone treatment there was: (i) no correlation between concentrations of progesterone and LH within days; (ii) a significant negative correlation between progesterone and days (P less than 0.01) and also between progesterone and LH over days (P less than 0.01); (iii) the overall correlation co-efficient between LH and days was positive (P less than 0.05). The amplitude of LH or FSH episodes was not affected as progesterone concentrations declined during PRID treatment, but the number of LH (but not FSH) episodes was increased (p less than 0.01). After PRID removal, the amplitude of both LH and FSH episodes increased (P less than 0.01). We suggest that progesterone is part of a negative feedback complex on LH secretion in cattle and that this effect is apparently mediated through frequency of episodic LH release.     

Structural and functional responses of wheat mitochondrial membranes to growth at low temperatures

The responses in membrane lipid composition, structure, and function of four cultivars of wheat (Triticum aestivum L.) to growth at low temperature have been investigated. Marked growth temperature-dependent alterations in the fatty acid composition and unsaturation of the mitochondrial phospholipids correlate with changes in respiratory activity in all the varieties. Parameters such as the respiratory control ratio and the phosphorylative efficiency decrease in cold-adapted seedlings. Three temperature-dependent structural transitions were identified in the mitochondria by the spin-labeling method. The structural transitions occur at lower temperatures in the cold-grown material. The shift in one transition appears to be quantitatively greater in the winter hardy varieties. Cold-induced changes in all of the other measured parameters were indistinguishable in hardy and nonhardy varieties. The results indicate major involvement of the phospholipid matrix in cold acclimation. A link between cold acclimation and winter survival may exist involving the structural and functional modifications in membrane structure which occur during acclimation.