Enzyme kinetics of a highly purified mitochondrial creatine kinase in comparison with cytosolic forms

Summary Mitochondrial creatine kinase (CK) purified from canine myocardium showed a single protein band on SDS-PAGE and was free of MMCK. Its amino acid composition was different than MMCK or BBCK and did not react to antiserum to MMCK or BBCK. Using purified mitochondrial, MM and BBCK, the velocity of reaction (V) was estimated for creatine phosphate (CP), creatine (C), adenosine triphosphate (ATP) and adenosine diphosphate (ADP) over a wide range of concentrations including those at V_max. The values for K_m (mM/L) derived from Lineweaver-Burke plots are shown: The affinity of mitochondrial CK for C is much greater than MMCK which is compatible with the energy shuttle hypothesis, namely ATP is converted by mitochondrial CK to CP, and then diffuses to the myofibril for conversion to ATP for utilization.     

Growth and mortality of zebra fish, Brachydanio rerio (Hamilton Buchanan), maintained at two temperatures and on two diets

One hundred zebra fish per tank were maintained for 112 days at 24°C or 28°C in glass aquaria and fed a diet of flaked food made without cellulose (13.45 kJ g^?1, metabolizable energy, Type A) or with cellulose (8.71 kJ g^?1, metabolizable energy, Type B). Each experimental condition was repeated in triplicate (12 tanks). The weight of food given daily to the fish was based on daily records of survivors (from which mortality rates were calculated) and wet wt of fish (measured every 14 days) in each tank. All fish were fed with the same weight of food per day and the quantity of energy in the food in excess of standard metabolism (as a proportion of SM) was approximately 0–5 for fish maintained at 28°C and fed food B, 1–0 for fish maintained at 24°C and fed food B, 1–5 for fish maintained at 28°C and fed food A, and 2.2 for fish maintained at 24°C and fed food A. Non-ionized ammonia, nitrite and nitrate nitrogen in the tanks did not reach toxic levels although there was an increase in total ammonium nitrogen in one tank and a subsequent heavy mortality. It was assumed that this was caused by the build up of pathogenic bacteria. Apart from this tank, mortality was highest in tanks at 28°C with fish fed food A and second highest in tanks at 24°C with fish fed on the same diet. Growth was measured in units of length, wet and dry weights, carbon and energy. There was a good correlation (P < 0.001) between carbon (mgC mg^?1) and calorific (J mg^?1) values and a conversion factor of 46.2 J (mgC)^?1 was derived. Fish maintained at 24°C and fed food A had the highest rates of growth both in weight and in energy value per unit weight. Fish fed the same diet but kept at 28°C had the lowest growth rates. Both these groups of fish had the highest coefficients of variation in wet weights which increased during the experiment, indicating an increase in interaction within the tanks. There was agreement between the energy value of fish sampled for growth and a condition factor based on the length-weight relationships of fish remaining in the tanks. A correlation (P < 0.05) was found between instantaneous mortality and growth rates for fish between tanks when those maintained at 28°C and fed on food A were ignored.     

Human alpha- and beta-CGRP and rat alpha-CGRP are coronary vasodilators in the rat

The effects of the recently described human alpha-calcitonin gene-related peptide (CGRP), human beta-CGRP and rat alpha-CGRP have been compared with those of the vasodilator sodium nitroprusside, on the rat and rabbit isolated heart. Hearts were perfused at constant flow and [Arg8]-vasopressin was used to increase coronary perfusion pressure. In the rat heart, the order of potency for evoking cumulative dose-dependent falls in perfusion pressure was human beta-CGRP greater than rat alpha-CGRP greater than human alpha-CGRP greater than sodium nitroprusside. In the same preparations the three CGRPs (but not sodium nitroprusside) elicited cumulative dose-related increases in heart rate. In the rabbit heart the order of potency for vasodilatation was rat alpha-CGRP greater than human alpha-CGRP greater than sodium nitroprusside. In marked contrast to results from the rat, neither rat alpha-CGRP nor human alpha-CGRP altered heart rate in the rabbit isolated heart. These results show that human alpha- and beta-CGRP and rat alpha-CGRP are vasodilators in the coronary vasculature, but that there is species variation as CGRP had a positive chronotropic effect in the rat heart but not in the rabbit heart.     

MORPHOLOGICAL VARIABILITY IN POPULATIONS OF CHRYSOPHYCEAN RESTING CYSTS. I. GENETIC (INTERCLONAL) AND ENCYSTMENT TEMPERATURE EFFECTS ON MORPHOLOGY1

Many chrysophycean species produce resting cysts (statospores) with purportedly species-specific morphology. I investigated variation in the cyst morphology of a single species that may result from genetic differences among the vegetative clones involved and from variation in the temperature of the environment during cyst development. Populations of Dinobryon cylindricum Imhof cysts were produced under defined conditions in vitro and then sampled for morphological analysis based on SEM micrographs. Morphological data is presented and then used in a multivariate discriminant analysis to determine the utility of each morphological character in distinguishing the six populations studied. Results suggest that some features of cyst morphology (i.e. cyst diameter) are invariant among the populations, while other features show distinctive variation. The density of spines covering the cyst body as well as the morphology of those spines appear correlated to the specific clones involved, and thus may represent useful phenotypic genetic markers. The length and definition of both the spines and the cyst collar, on the other hand, are markedly influenced by encystment temperature. The implications of these findings for paleoecological studies is discussed.     

Action of two juvenile hormone antagonists on lepidopteran epidermis

The juvenile hormone antagonist ETB (ethyl-4-2(t-butylcarbonyloxy)-butoxybenzoate) caused formation of precocious larval-pupal intermediates after the 4th (penultimate)-larval instar of the tobacco hornworm, Manduca sexta, when 50 μg were applied to any 3rd stage larvae or to 4th stage larvae within 12 hr after ecdysis. This dose was most effective within 12 hr after ecdysis to the 3rd stage. In the black mutant larval assay for juvenile hormone, ETB had activity, 0.75 μg per larva giving half-maximal score. In vitro ETB acted as a juvenile hormone to prevent the ecdysteroid-induced change in commitment at concentrations above 0.1 μg/ml with an ED₅₀ at 2.8 μg/ml and as a partial juvenile hormone antagonist to 0.1 μg/ml juvenile hormone I at concentrations between 10^?3 and 10^?2 μg/ml. By contrast, EMD (ethyl-E-3-methyl-2-dodecenoate) had little juvenile hormone-like activity in vitro up to its limits of solubility (100 μg/ml) and exhibited sporadic partial juvenile hormone antagonistic activity in vitro at concentrations between 1 and 100 μg/ml. Since these concentrations were 10–1000 times that of juvenile hormone I in the medium, EMD apparently is not an efficient competitor.     

Substitution of lysine for threonine at position 100 in human carbonic anhydrase Id Michigan

The position of the amino acid substitution in the human red cell carbonic anhydrase I variant, CA Id Michigan, has been determined by sequence analysis of the altered tryptic peptide. The threonine to lysine substitution was found to be located at position 100, and is expressed as CA I^{100 Lys}.