Influence of body weight/age on the food intake,growth and conversion efficiency of Gambusia affinis

On a restricted food supply of Tubifex tubifex (2h/day) the fish consumed only 69.5 mg dry food/g live fish/day and grew only to 25.6 mg live body weight by the 30th day of it's age, while those fed ad libitum consumed 94.7 mg/g/day and attained a body weight of 125.9 mg live weight by the 30th day. Conversion efficiency (K₁) averaged to 8.3% in the former and 14.1% in the latter series, throughout the 30 day experimental period.Feeding rate of the test series fed individually and at ad libitum, decreased from 780 mg dry food/g dry fish/day for a fish weighing 4 mg dry weight to 180 mg/g/ day for one weighing 41 mg, exhibiting a log-log relationship to the body weight (Y = 2.20140–0.5639 X). Growth rate also decreased as a log-log function with increasing body weight (Y = 1.2309–0.4384 X).     

Machinery to support genome segment inversion exists in a herpesvirus which does not naturally contain invertible elements

In many herpesviruses, genome segments flanked by inverted repeats invert during DNA replication. It is not known whether this inversion is a consequence of an inherently recombinagenic replicative mechanism common to all herpesviruses or whether the replication enzymes of viruses with invertible segments have specifically evolved additional enzymatic activities to drive inversion. By artificially inserting a fusion of terminal sequences into the genome of a virus which normally lacks invertible elements (murine cytomegalovirus), we created a genome composed of long and short segments flanked by 1,359- and 543-bp inverted repeats. Analysis of genomic DNA from this virus revealed that inversion of both segments generates equimolar amounts of four isomers during the viral propagation necessary to produce DNA for analysis from a single viral particle. We conclude that a herpesvirus which naturally lacks invertible elements is able to support efficient segment inversion. Thus, the potential to invert is probably inherent in the replication machinery of all herpesviruses, irrespective of genome structure, and therefore genomes with invertible elements could have evolved simply by acquisition of inverted repeats and without concomitant evolution of enzymatic activities to mediate inversion. Furthermore, the recombinagenicity of herpesvirus DNA replication must have some importance independent of genome segment inversion.     

Biochemical Changes in Nodules of Vigna mungo (L.) During Vegetative and Reproductive Stages of Plant Growth in the Field

Nitrogenase activity in root nodules of Vigna mungo L. attaineda peak at the flowering stage and declined thereafter. Levelsof soluble proteins, particularly leghaemoglobin, and ratesof protein and RNA synthesis declined with nitrogenase. Activitiesof protease and RNase increased with the ageing or nodules.Carbohydrate utilization, sugar levels and ATP were maximumat the early pod stage and gradually declined with age. Theseinterrelated changes point to a loss of nitrogenase activityas the first indicator of nodule senescence that is linked withflowering. Later, losses of proteins, total sugar and ATP wererelated to increased RNase and protease activity and decreasedhexokinase and to a loss in capacity to incorporate amino acidinto protein.Copyright 1993, 1999 Academic Press Vigna mungo (L.), senescene, nitrogenase, leghaemoglobin, field experiments     

Crustacean egg size as an indicator of egg fat/protein reserves

The metabolic properties of crustacean eggs and the major organic reserve utilized for embryogenesis are dependent on the egg-size correlated deposition of fat/protein in the yolk.     

Expression of human chorionic gonadotropin uniformly labeled with NMR isotpes in Chinese hamster ovary cells: An advance toward rapid determination of glycoprotein structures

Summary Most secreted eukaryotic proteins are modified by glycosylation, and it has been difficult to solve their structures by crystallographic or NMR techniques because of problems posed by the presence of the carbohydrate. The structure of a chemically deglycosylated form of the human pregnancy hormone, human chorionic gonadotropin (hCG), has been solved by crystallographic methods. Since chemical deglycosylation may have induced changes in the structure, and since it is known that deglycosylated hCG is biologically inactive, the crystallographic structure requires confirmation by NMR techniques. Also, it has not been possible to determine the structures of the isolated subunits, nor the nature of interactions between the carbohydrate side chains and the protein back bone by crystallographic methods. Structural information via NMR techniques can be obtained from proteins in solution if they can be uniformly labeled with ¹³C and ¹⁵N isotopes. We report the first such uniform labeling of a glycoprotein using a universal ¹³C-and ¹⁵N-labeling medium to express ¹³C, ¹⁵N-labeled hCG, suitable for solving the structure in solution of the native, biologically active form of hCG as well as that of its free subunits. The ¹³C, ¹⁵N-labeled recombinant hCG and its separated subunits are shown to be nearly identical to urinary hCG reference preparations on the basis of protein chemical studies, immunochemistry, biological activity, and the capability of isolated hormone subunits to recombine to form biologically active hormone. Mass spectrometric analysis and preliminary NMR studies indicate that the isotopic labeling is uniform and greater than 90% after only two growth passages in the labeling media. One unexpected finding during subunit purification was that lyophilization of glycoproteins from trifluoroacetic acid HPLC buffers may result in the loss of a significant portion of sialic acid.To whom correspondence should be addressed.     

Increased enzyme secretion by 2-deoxy-d-glucose in presence of succinate by suppression of metabolic enzymes in Termitomyces clypeatus

Regulatory mode of secretion of proteins was detected for the industrial glycosidase, cellobiase, under secreting conditions (in presence of TCA cycle intermediates like succinate etc.) in the filamentous fungus Termitomyces clypeatus. The titers of key metabolic enzymes were investigated under secreting and non-secreting conditions of growth and compared to the corresponding production of intra and extracellular levels of cellobiase. Results were compared in presence of 2-deoxy-d-glucose, a potent glycosylation inhibitor in the secreting media. Inclusion of 2-deoxy-d-glucose in presence of succinate caused about 10 to 100 times decrease in titers of the metabolic enzymes hexokinase, fructose-1,6-bisphosphatase, isocitrate lyase and malate dehydrogenase leading to increased secretion of cellobiase by more than 100 times. The intracellular concentration of cAMP (86-fold decrease in presence of 2-deoxy-d-glucose under secreting conditions) and turnover rate of proteins also dropped significantly. In this suppressed metabolic state, a 10-fold increase in the titer of the secreted cellobiase was noticed. The results indicated elucidation of carbon catabolite repression like phenomenon in the fungus under secreting conditions which was more pronounced by 2-deoxy-d-glucose. The interdependence between secretion and regulation of metabolic enzymes will help in better understanding of the physiology of these highly adapted organisms for increasing their secretion potential of glycosidases like cellobiase with high industrial value.