Complex CIS-Acting Regulators and Locus Structure of Drosophila Tissue-Specific Adh Variants

Diverse patterns of tissue-specific expression of alcohol dehydrogenase (ADH) among species of the grimshawi subgroup of Hawaiian picture-winged Drosophila suggests control by complex or multiple, independently acting regulatory elements. These elements act by controlling Adh mRNA accumulation in individual tissue types. Restriction mapping of the Adh loci from these species reveals several insertion/deletion differences, one of which lies just outside the 5' end of the structural sequences and correlates with differences in larval patterns of ADH expression. No tissue-specific rearrangement of Adh sequences was observed.     

The role of the metal ion in the mechanism of the K+-activated aldehyde dehydrogenase of Saccharomyces cerevisiae.

The effect of K+ on assays of the enzyme was studied and it appears that the activation occurs slowly by a two-step process. Kinetic measurements suggest that the enzyme-catalysed reaction can proceed slowly (0.4%) in the complete absence of K+. The enzyme exhibits a K+-activated esterase activity, which is further activated by NAD+ or NADH. Stopped-flow studies indicated that the principal effect of K+ on the dehydrogenase reaction is to accelerate a step (possibly acyl-enzyme hydrolysis) associated with a fluorescence and small absorbance transient that occurs after hydride transfer and before NADH dissociation from the terminal E-NADH complex. The variation of activity of the enzyme with pH was studied. An enzyme group with pKa approx. 7.1 apparently promotes enzyme activity when in its alkaline form.     

Freezing damage to isolated tomato fruit mitochondria as modified by cryoprotective agents and storage temperature

Isolated tomato (Lycopersicon esculentum var. Kc 146) fruit mitochondria could be stored successfully in the frozen state without a cryoprotective agent if the mitochondria were frozen quickly by immersion in liquid nitrogen and later thawed quickly at 30 C. Criteria of freezing damage were rate of respiration, adenosine diphosphate to oxygen ratio, and respiratory control ratio. Marked reduction in respiration and loss of respiratory control occurred when mitochondria were transferred from liquid nitrogen to −5, −10, or −18 C for 15 minutes prior to thawing at 30 C. Dimethylsulfoxide (5%) prevented freezing damage when mitochondria were incubated at −5 C but did not prevent freezing damage at −10 or −18 C. Isolated tomato mitochondria show promise as a model system for studying the nature of freezing damage and the mode of action of cryo-protective agents.     

Use of Dexamethasone in Cerebral Malaria

In a man who had been in a coma for 24 hours with cerebral malaria dexamethasone had a dramatic, and probably life-saving, effect. We believe dexamethasone should be given routinely, together with antimalarial therapy, to patients with cerebral malaria.     

The ribosome cycle,nucleoli, and cytoplasmic nucleoloids in the meiocytes ofLilium

Summary During the meiotic prophase in pollen mother cells ofLilium species the ribosome population of the cytoplasm diminishes to a minimum reached during the diplotene-diakinesis interval. This fall is associated with a drop in the ribonuclease-extractable RNA present in whole, fixed cells. A rise in RNA and in number of observable ribosomes follows during the meiotic mitoses and in the very early life of the spores. Bodies with the characteristics of nucleoli—here termed nucleoloids—are released into the cytoplasm at anaphase I and anaphase II, and it is suggested that these may represent the main mechanism through which the ribosome population of the cytoplasm is restored after the prophase elimination.     

Agar Medium for the Selective Enumeration of Coagulase-positive Staphylococcus from the Rat Alimentary Tract

A selective agar medium (medium J(1)) is proposed for the quantitative enumeration of egg yolk-positive (EYP) and egg yolk-negative (EYN) Staphylococcus pyogenes from the digestive tract and feces of the rat. This medium, buffered at pH 5.0, is composed of acid casein hydrolysate and yeast extract with 7.5% sodium chloride, 1.6% sodium pyruvate, 0.0008% 2,3,5-triphenyltetrazolium chloride (TTC), and 6% egg yolk emulsion. Inoculation is by the pour-plate method and incubation is at 38 C in a water-jacketed incubator for 36 hr. Colonies of S. pyogenes reduce TTC; EYP strains are surrounded by a halo of opacity; and EYN strains may be surrounded by a red halo, but no opacity. Small, white colonies of S. epidermidis may develop, but Micrococcus, and all other groups of Staphylococcus recognized in the rat intestinal flora, are inhibited. Other bacterial genera, notably Bacillus, Corynebacterium, Proteus, and Streptococcus, are also inhibited.     

Thyroid function after subtotal thyroidectomy for hyperthyroidism.

Among 76 patients who had had a subtotal thyroidectomy for hyperthyroidism from one to seven years previously recurrent hyperthyroidism was found in three and hypothyroidism in 13. The remaining 60 subjects were clinically euthyroid but a raised level of serum thyroid-stimulating hormone (TSH; greater than 5-0 mu U/ml) was found in 39. Analysis of the data showed that their serum thyroxine was significantly lower than in the subjects with a normal TSH. The serum triiodothyronine (T-3) was similar in both groups. It is concluded that subjects with a raised TSH remain clinically euthyroid by maintaining a normal serum T-3 concentration. There was no evidence of any long-term progressive deterioration of thyroid function after subtotal thyroidectomy.     

Kinetic properties of aldehyde dehydrogenase from sheep liver mitochondria.

The kinetics of the NAD+-dependent oxidation of aldehydes, catalysed by aldehyde dehydrogenase purified from sheep liver mitochondria, were studied in detail. Lag phases were observed in the assays, the length of which were dependent on the enzyme concentration. The measured rates after the lag phase was over were directly proportional to the enzyme concentration. If enzyme was preincubated with NAD+, the lag phase was eliminated. Double-reciprocal plots with aldehyde as the variable substrate were non-linear, showing marked substrate activation. With NAD+ as the variable substrate, double-reciprocal plots were linear, and apparently parallel. Double-reciprocal plots with enzyme modified with disulfiram (tetraethylthiuram disulphide) or iodoacetamide, such that at pH 8.0 the activity was decreased to 50% of the control value, showed no substrate activation, and the plots were linear. At pH 7.0, the kinetic parameters Vmax. and Km NAD+- for the oxidation of acetaldehyde and butyraldehyde by the native enzyme are almost identical. Formaldehyde and propionaldehyde show the same apparent maximum rate. Aldehyde dehydrogenase is able to catalyse the hydrolysis of p-nitrophenyl esters. This esterase activity was stimulated by both NAD+ and NADH, the maximum rate for the NAD+ stimulated esterase reaction being roughly equal to the maximum rate for the oxidation of aldehydes. The mechanistic implications of the above behaviour are discussed.