An improved technique for sequential R-, Q- and C-banding of bone marrow chromosomes

A procedure for the sequential staining of metaphase cells to provide R-banding, Q-banding and C-banding of chromosomes is described. The combined techniques allow more precise identification of marker chromosomes and more accurate definition of breakpoints in the consistent chromosome translocations that are associated with certain human malignancies.     

Purification and characterization of the androgen receptor from rat ventral prostate

The androgen receptor has been purified from rat ventral prostate cytosol by a combination of differential DNA-Sepharose 4B chromatography and testosterone 17 beta-hemisuccinyl-3,3'-diaminodipropylamine-Sepharose 4B affinity chromatography. Approximately 8 micrograms of protein was obtained from 38 g of rat ventral prostate, with a yield of 24%. The receptor was purified approximately 120 000-fold. Silver nitrate staining of a sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel revealed a major polypeptide band migrating at 86 000 daltons. Affinity labeling of a partially purified receptor preparation with either 17-hydroxy-17 alpha-[3H]methyl-4,9,11-estratrien-3-one or 17 beta-hydroxy-[1,2,4,5,6,7,16,17-3H8]-5 alpha-androstan-3-one 17-(2-bromoacetate) produced a major band of radioactivity migrating at 86 000 daltons on a NaDodSO4 gel. Under nondenaturing conditions, a Mr of 85 000 was determined by gel filtration (42 A) and sucrose gradient sedimentation analysis (4.5 S). The purified receptor had an isoelectric point of 6.3 [3H]-4,5 alpha-Dihydrotestosterone, bound to the purified receptor, was displaced with 4,5 alpha-dihydrotestosterone greater than testosterone much greater than progesterone greater than 5 alpha-androstane-3 alpha, 17 beta-diol greater than 17 beta-estradiol greater than cortisol. A number of physicochemical properties of the purified receptor were similar to those of the receptor in crude cytosol.     

Reactions of ferrioxamine and desferrioxamine with the hydroxyl radical

Both ferrioxamine and desferrioxamine react with the hydroxyl radical with a second order rate constant equal to 1.3 × 10¹⁰ M^?1 s^?1. Conditions for the use of desferrioxamine as a probe for the role of iron salts in the formation of hydroxyl radicals in biochemical systems are discussed.     

Observations on effects of feeding level on growth and reproduction in haddock, Melanogrammus aeglefinus (L.) in captivity

Individual haddock, Melanogrammus aeglefinus (L.) were maintained at different feeding levels in an aquarium from November until the completion of spawning. The mean duration of spawning was 33.2 days (range 19–59) during which an average of 16.6 (range 10–25) batches of eggs were produced. The size and dry weight of the eggs declined during the spawning period. Egg production and feeding level were correlated positively. There was some suggestion that when female haddock received low rations (< 5 kcal day^-1) a lower proportion spawned, and the dry weights of the eggs were lower compared with females on high rations (> 13 kcal day ^-1). The relation between daily growth in wet weight, g, and daily surplus energy intake, kcal, was: G =(0.295 E )— 0.328. When food energy is restricted, haddock appear to achieve a balance between somatic growth and reproduction.     

Isolation of a yeast-lyzing Arthrobacter species and the production of the lytic enzyme complex in batch and continuous-flow fermentors

A gram-negative bacterium strongly lytic toward living cells of the food yeast Saccharomyces fragilis was isolated by continuous-flow enrichment from compost. The organism was identified as a species of Arthrobacter. The extracellular lytic enzyme complex produced by this bacterium contained β-1,3-glucanase, mannan mannohydrolase, and proteolytic activities. The polysaccharases were inducible by whole yeast cells. In chemostat cultures on chemically defined media, synthesis of the polysaccharases was very slight and only detectable at dilution rates below 0.02 hr^?1. Enzyme production in defined media was not solely dependent on growth rate but also was influenced by the growth limiting substrate and the culture history. The production of individual depolymerases and of the lytic activity was studied in batch and chemostat cultures containing yeast as the limiting substrate. The maximum specific growth rate of the Arthrobacter under these conditions was 0.22 hr^?1. β-1,3-Glucanase and proteolytic activities were synthesized by exponentially growing bacteria but maximum lytic titers did not develop until the specific growth rate was declining, at which time mannan mannohydrolase syntheses was induced. In yeast limited chemostats polysaccharase syntheses were greatest at the lowest dilution rates examined, namely 0.02 hr^?1. Further optimization of enzyme production was achieved by feeding the Arthrobacter culture to a second-stage chemostat. A comparison of lytic enzyme productivities in batch and chemostat cultures has been made.     

Piscine predation on juvenile fishes on a Scottish sandy beach

Predation by larger fishes is a major cause of mortality for the populations of juvenile fishes on a sandy beach on the west coast of Scotland. Predation was concentrated on the most numerous species (0-group Pleuronectes platessa ) in June but with the decline in numbers and growth in size of this species, the fish predators had changed their diet in August to feed principally on small sandeels (Ammodytidae). Six major predatory species had taken 95 per cent of all fish found in predators' stomachs and could be roughly divided into three categories according to their piscivorous tendencies and their abundance. (1) Common species in which fishes formed the main item of their diet ( Gadus morhua ). (2) Species in which fishes formed only a relatively minor constituent of the diet but which, by virtue of their abundance, potentially had a significant predatory impact (I-group P. platessa ). (3) less abundant species whose diet consisted principally of fishes. The intensity of predation was generally greatest at night for G. morhua but during the day for I-group P. platessa. The distributions and movements of the dominant prey species can be interpreted in the light of their predator-prey relationships. The most common benthic species, 0-group P. platessa , move off -and onshore with the ebb and flow of the tides and concentrate in shallow water at night. Such movements would have the effect of maintaining the majority of the population in depths where predation pressure from nocturnally onshore-migrating gadoids and other predators is likely to be minimized.     

Partition coefficients of substrates and products and solvent selection for biocatalysis under nearly anhydrous conditions

The effect of solvent on the activity of mushroom tyrosinase toward three substrates was studied at a constant water activity of either 0.74 or 0.86. No simple correlation was observed between enzyme activity and log P, but partition coefficients of substrate (P(s)) and product (P(p)) gave systematic relations with enzyme activity. When initial reaction rates were considered, there was a bellshaped relationship between enzyme activity and P(s) with an optimal P(s) for each substrate. This can be explained by assuming that the solvent affected the enzyme activity primarily by affecting the substrate concentration in the aqueous layer around the catalyst where the enzymic reaction occurs. When long-term reaction rates were considered, a high P(p)/P(s) ratio was consistent with preservation of enzyme activity. (c) 1994 John Wiley & Sons, Inc.     

Purification and characterization of NADP-specific alcohol dehydrogenase and glutamate dehydrogenase from the hyperthermophilic archaeon Thermococcus litoralis.

Thermococcus litoralis is a strictly anaerobic archaeon that grows at temperatures up to 98 degrees C by fermenting peptides. Little is known about the primary metabolic pathways of this organism and, in particular, the role of enzymes that are dependent on thermolabile nicotinamide nucleotides. In this paper we show that the cytoplasmic fraction of cell extracts contained NADP-specific glutamate dehydrogenase (GDH) and NADP-specific alcohol dehydrogenase (ADH) activities, neither of which utilized NAD as a cofactor. The GDH is composed of identical subunits having an M(r) of 45,000 and had an optimal pH and optimal temperature for glutamate oxidation of 8.0 and > 95 degrees C, respectively. Potassium phosphate (60 mM), KCl (300 mM), and NaCl (300 mM) each stimulated the rate of glutamate oxidation activity between two- and threefold. For glutamate oxidation the apparent Km values at 80 degrees C for glutamate and NADP were 0.22 and 0.029 mM, respectively, and for 2-ketoglutarate reduction the apparent Km values for 2-ketoglutarate, NADPH, and NH4+ were 0.16, 0.14, and 0.63 mM, respectively. This enzyme is the first NADP-specific GDH purified form a hyperthermophilic organism. T. litoralis ADH is a tetrameric protein composed of identical subunits having an M(r) of 48,000; the optimal pH and optimal temperature for ethanol oxidation were 8.8 and 80 degrees C, respectively. In contrast to GDH activity, potassium phosphate (60 mM), KCl (0.1 M), and NaCl (0.3 M) inhibited ADH activity, whereas (NH4)2SO4 (0.1 M) had a slight stimulating effect. This enzyme exhibited broad substrate specificity for primary alcohols, but secondary alcohols were not oxidized.(ABSTRACT TRUNCATED AT 250 WORDS)     

Skeletal signs of activity in the Italian metal ages: Methodological and interpretative notes

In recent years anthropologists have made much progress in understanding ancient activities from skeletal remains. In this paper, material from the Iron Age cemetery at Pontecagnano (VII-IV century BC) is used to illustrate activity-related traits of eight basic categories: