Analysis in vivo of translational mutants of the rIIB cistron of bacteriophage T4

We have mapped the mutants isolated by Nelson et al. (1981) that reduce the amount of rIIB protein synthesized during bacteriophage T4 infection of Escherichia coli B and characterized their rIIB expression in vivo. These mutants fall into four distinct groups in terms of mapping and phenotype. We have located the probable site of each mutation on the DNA sequence. We have also analyzed a number of other mutations near the initiating AUG of rIIB with respect to their rIIB expression. In some of these mutants, ribosomal recognition of the wild-type initiating AUG is precluded and so initiation occurs at a different AUG, which, in some instances, we have identified.     

An exceptional amino acid replacement on the distal side of the iron atom in proboscidean myoglobin

Summary Amino acid sequence determination of elephant myoglobin revealed the presence of the unusual substitution E7 His Gln. Stereochemical analyses suggest that the most suitable residue which can functionally substitute for His at this position in vertebrate globins is Gln. Physiochemical studies imply that the slower rate of autooxidation of elephant myoglobin is the result of this substitution which may confer some selective advantage on the species. Comparative sequence data of paenungulate myoglobins suggest that the His Gln mutation probably occurred in an ancestor of Elephantinae.     

Characterization of Citrus Rootstock Responses to Tylenchulus semipenetrans (Cobb)

Citrus rootstocks which significantly limited the reproduction of Tylenchulus semipenetrans (Cobb) "Citrus" and "Poncirus" biotypes responded to infection by producing a hypersensitive-type response in the root hypodermis, wound periderm and/or cavities in the root cortex, and/or abnormal vacuoles in nurse cell cytoplasm. Rootstocks which limited nematode reproduction also had significantly fewer nematodes in the rhizoplane within 8 d of inoculation than did rootstocks which did not limit reproduction. Germplasm sources of the cellular responses which limited citrus nematode reproduction were identified.     

Identification and Distribution of m- and p-Hydroxyphenylacetic Acids in the Brain of the Rat

The m and p isomers of hydroxyphenylacetic acid have been identified and quantitated in whole rat brain and in several regions using a capillary column high resolution gas chromatography–mass spectrometry procedure. Their concentrations were: for m-hydroxyphenylacetic acid (mean ± S.E., number of determinations in parentheses)—whole brain, 2.3 ± 0.3 ng/g (7); hypothalamus, 1.2 ± 0.3 ng/g (5); caudate nucleus, 5.5 ± 0.6 ng/g (5); brain stem, 1.8 ± 0.1 ngig (5); cerebellum, 1.2 ± 0.1 ng/g (5) and the “rest,” 1.7 ± 0.1 ng/g (5); and for p-hydroxyphenylacetic acid–whole brain, 10.6 ± 0.7 ng/g (7); hypothalamus, 4.5 ± 0.1 ng/g (4); caudate nucleus, 28.3 ±1.6 ng/g (5); brain stem, 8.6 ± 0.6 ng/g (5); cerebellum, 8.1 ± 0.4 ng/g (9, and the “rest,” 5.3 ± 0.5 ng/g (5). This heterogeneous distribution parallels closely that exhibited by their respective precursor amines, m- and p-tyramine.     

Narcotic antagonists attenuate drinking induced by water deprivation in a primate

Pure narcotic antagonists such as naloxone and naltrexone have consistently been shown to attenuate drinking in the rat after periods of water deprivation. One objective of this study was to extend observations to a primate species, the squirrel monkey. Whereas naloxone and naltrexone have a greater relative affinity for opiate receptors preferentially binding morphine and other opiate alkaloids than for those with high affinity for the endogenous opioid peptides, diprenorphine, another pure opiate antagonist, binds with equally high affinity to both receptor subtypes. Therefore, a second objective was to determine the actions of diprenorphine on drinking in water-deprived rats and squirrel monkeys and to compare the effects of this drug to those of naloxone and naltrexone. All three narcotic antagonists suppressed water consumption of monkeys and rats deprived of water for 18 and 24 hr, respectively. Diprenorphine was the most potent compound tested in both species, producing significant reductions in water consumption of monkeys and rats at systemic doses as low as 0.01 and 0.1 mg/kg respectively. Moreover, diprenorphine was the longest acting of the three drugs in the monkey. These results demonstrate that the narcotic antagonists attenuate drinking in primates as well as in rodents and support the hypothesis that these drugs reduce water intake by interrupting the activity of endogenous opioid pathways mediating drinking behavior.     

The effect of exhaustive exercise on expired pentane as a measure of in vivo lipid peroxidation in the rat

The concentrations of pentane and ethane in the expired breath of swimming rats were used to determine the possible occurrence of lipid peroxidation caused by strenuous exercise. Rats swum to exhaustion produced significantly more pentane but not ethane than did rats at rest. Rats fed a vitamin E-deficient diet produced slightly more pentane following exhaustive exercise than they did while at rest, but this increase was not statistically significant. Rats were also swum for prescribed lengths of time. Only rats that had swum for 20 or 40 min had significantly elevated concentrations of pentane in the breath. Rats swum for 10 or 30 min had elevated concentrations of pentane in the breath, but these increases were not statistically significant. These results suggest that lipid peroxidation is moderately increased following exhaustive exercise.     

Comparison of energy budgets for spruce budworm Choristoneura fumiferana (Clemens) on balsam fir and white spruce

Summary A determination was made of the differences in the utilization of energy by laboratory reared larval Choristoneura fumiferana fed either balsam fir or white spruce foliage. This enabled us to quantitatively measure the quality of these foliages vis a vis the spruce budworm.The larval strategy was to feed rapidly and develop quickly. Development time was longer on spruce than on fir. Total consumption was virtually identical on both foliage types though production was ca 20% greater on a white spruce diet. Calories/gram increased with insect development and with the development of balsam fir foliage but declined over time in white spruce. Assimilation efficiencies (A/C) were 33.9% to 40.2% while gross production efficiencies (P/C) were 9.5% to 13.3% and net production efficiencies (P/A) were 26.1% to 38.8%. The spruce fed insects gave higher values than those on fir, but all values were at the low end of the range for lepidopterous defoliators (possibly due to characteristics of conifer foliage).The enhanced growth found in spruce fed larvae contrasts the relationship between balsam fir and high insect densities. This is discussed together with the influence of spruce budworm on forest composition and the possible competitive benefits of spruce budworm induced tree mortality.Parts of this paper appeared in a Master's thesis (Koller 1978) submitted to the University of Maine     

Incorporation of label from root-applied N6[8-14C]furfiuryIadenine into the guanine nucleotide fraction of pea bud ribonucleic acid

The pattern of incorporation of label into the nucleotides of axillary bud ribonucleic acid was investigated in Pisum sativum L. cv. Meteor following the application of N ⁶[8-^I4C]furfuryladenine or of [8-¹⁴C]adenine to the root system of decapitated plants and to cultured excised buds. When N ⁶[8-¹⁴C]furifaryladenine was applied to the root system label was confined to the guanine nucleotide moiety of the axillary bud ribonucleic acid; label from [8-¹⁴C]adenine was incorporated preferentially into adenine nucleotide in the molar ratio adenine nucleotide/guanine nucleotide = 3.23. When isolated buds were incubated in media containing [8-¹⁴C]adenine or N ⁶[8-¹⁴C]furfuryladenine, label was incorporated into both purine moieties of the ribonucleic acid. However, the relative incorporation into the guanine nucleotide fraction was considerably greater for N ⁶[8-^I4C]furfuryladenine (adenine nucleotide/guanine nucleotide = 2.23) than for [8-¹⁴C]adenine (ratio = 4.67).
It was concluded that the pattern of metabolism of adenine to guanine and its incorporation into the guanine nucleotide moiety of pea axillary bud ribonucleic acid, is influenced by the presence of a substitution in the N ⁶ position of the adenine base.     

Synthesis and biological activity of brassinolide and its 22β,23β-isomer: Novel plant growth-promoting steroids

Brassinolide (2α,3α,22α, 23α-tetrahydroxy-24α-methyl-B-homo-7-oxa-5α-cholestan-6-one), a novel plant growth-promoting steroid isolated from rape pollen, and its hitherto unknown 22β, 23β-isomer were synthesized from a C-24 epimeric 60:40 mixture of 22-dehydrocampesterol (24α-methyl) and brassicasterol (24β-methyl) from oysters. The method of synthesis favored the formation of the 22β, 23β-isomer by better than 4:1. Comparative plant growth-promoting capabilities of brassinolide, both natural and synthetic, and its three side chain $\text{cis}$-glycolic isomers in the bean second internode bioassay showed that the natural and synthetic brassinolides were equally active and caused splitting of the internode at the 0.1 μg level. The least active was the 22β,23β-isomer of brassinolide. The isomers with the 22α, 23α and 24β, and the 22β, 23β and 24β configurations were highly active and were required at about 10 times the concentration of brassinolide to cause the same physiological response. In the bean first internode bioassay, an auxin-induced growth test system which employs isolated bean plant segments, the isomer with 22β, 23β and 24β configuration caused a greater response than brassinolide. Two of the four tetrahydroxy ketones obtained in the synthesis of the isomers were also active in both assays.     

Fluorescence photobleaching recovery measurements reveal differences in envelopment of sindbis and vesicular stomatitis viruses

Fluorescence photobleaching recovery (FPR) measurements of virus glycoproteins on the surfaces of cells infected with vesicular stomatitis virus (VSV) and Sindbis virus showed that the VSV glycoprotein (G) remained mobile throughout the infectious cycle, whereas Sindbis virus glycoproteins (E1, E2) were partially mobile early after infection and immobile at later times when greater amounts of these proteins were on the cell surface. A highly mobile fraction of Sindbis virus glycoproteins was detected throughout the replication cycle of a temperature-sensitive mutant unable to form virus particles. Thus immobilization of E1 and E2 was the result of increasing surface glycoprotein concentrations and virus budding. Together with other data, which included the detection of E1 and E2 in particles as soon as these proteins were transported to the cell surface, the FPR results suggest that Sindbis virus assembly initiates on intracellular vesicles, where glycoproteins aggregate and bind nucleocapsids. In contrast, our FPR data on VSV support a model previously suggested by others, in which a small fraction of cell-surface G is immobilized into budding sites formed by interactions with virus matrix and nucleoproteins. FPR measurements also provide direct evidence for strong interactions between E1 and E2, as well as between E1 and PE2, the precursor form of E2.