Polyproline binding is an essential function of human profilin in yeast.

Structural analysis of human profilin has revealed two tryptophan residues, W3 and W31, which interact with polyproline. The codons for these residues were mutated to encode phenylalanine and the mutant proteins overexpressed in Eschericia coli. The isolated proteins were diminished in their ability to bind polyproline, whereas phosphatidylinositol 4,5-bisphosphate (PIP2) binding remained unchanged. In many strains of Saccharomyces cerevisiae, disruption of the gene encoding profilin, PFY1, is lethal. It was found that expression of the gene for human profilin is capable of suppressing this lethality. The polyproline-binding mutant alleles of the human gene were cloned into various yeast expression vectors. Each of the mutant genes resulted in suppression of the lethality of pfy1Delta. It was observed that the mutant protein expression levels paralleled the growth rates of the strains. The severity of various morphological abnormalities of the strains was also attenuated with increased protein levels, suggesting that profilin polyproline-binding mutations are deleterious to cell growth unless overexpressed. Both tryptophan mutations were combined to give a third mutant allele that was found both unable to bind polyproline and to suppress the lethality of a pfy1 deletion. Immunoprecipitation experiments suggested that the mutants were unaltered in their affinity for actin and PIP2. These data strongly suggest that polyproline binding is an essential function of profilin.     

A stable isotope trace method to measure silicic acid uptake by marine phytoplankton.

A tracer method is described that uses the stable isotope ³⁰Si to measure rates of silicic acid uptake by diatom cultures and natural populations of marine phytoplankton. The method involves (i) incubation of organisms requiring silicic acid for growth in the presence of ³⁰Si-labeled silicic acid, (ii) collection of the resulting particulate silicon, (iii) conversion of the particulate silicon to BaSiF₆, (iv) determination of the ³⁰Si content of BaSiF₆ by solid sample mass spectrometry, and (v) calculation of the uptake rate from the ³⁰Si enrichment of the particulate matter during the incubation. The maximum overall error in the uptake rate measurement is ±10%.     

The acidic amino acid transport system of the baby hamster kidney cell line BHK21-C13

The uptake of L-glutamate into BHK21-C13 cells in culture has been studied. This amino acid appears to be transported via a relatively high affinity, low capacity, Na+-dependent transport system capable of the rapid accumulation of substrate amino acids. Kinetic studies of the inhibition of L-glutamate uptake has provided information as to the substrate and the molecular configuration required for transport via the glutamate transport system. This system exhibited marked substrate specificity and was only capable of transporting L-glutamate and aspartate and certain closely related acidic amino acid analogues.     

Howardula dominicki n. sp. Infesting the Tobacco Flea Bettle in North Carolina

Howardula dominicki n. sp. is described from specimens collected from the tobacco flea beetle, Epitrix hirtipennis (Melsheimer), at Oxford, North Carolina , and is distinguished from other members of the genus . Parasitism by H. dominicki sterilized female flea beetles and often led to the death of larvae.     

Purified viral neuraminidase vaccine to control influenza.

The control of influenza by immunoprophylaxis is difficult because of the antigenic mutability of the influenza virus and the unpredictability of its epidemiologic behaviour. The inactivated whole-virus vaccine currently used is not ideal. Vaccination with pure neuraminidase is suggested. The induced antineuraminidase antibody will restrict viral invasion. Mild illness may or may not occur. On subsequent exposure to influenza virus the individual will produce antihemagglutinin and antineuraminidase antibodies and will be resistant to both infection and illness. Since antigenic changes are less frequent in the viral neuraminidase than in the viral hemagglutinin, the vaccine would be usable for longer periods than the presently used inactivated whole-virus vaccine.     

Effect of change in particle number on pulmonary clearance of aerosolized 99mTc-DTPA

Pulmonary clearance (PCl) of inhaled aerosolized 99mTc-diethylenetriamine pentaacetic acid (DTPA) across the alveolocapillary membrane is diffusion limited. Therefore, if the mixing of the 99mTc-DTPA in the aqueous hypophase underlying surfactant is slow or incomplete or if there were no hypophase, an increase in the alveolar surface area occupied by 99mTc-DTPA particles would increase the absorption rate. The aim of this study was to examine whether there is an effect on PCl of changing the number of inhaled particles. The change in particle number was accomplished by a setup of four parallel jet nebulizers feeding a central delivery chamber of 400 cm3. We performed two kinds of experiments in eight healthy nonsmokers between 28 and 52 yr of age. In the first experiment, 99mTc-DTPA in saline was nebulized in one nebulizer, while saline was nebulized in the other three. In the second experiment the number of inhaled particles containing 99mTc-DTPA was increased by a factor of four by nebulizing 99mTc-DTPA in saline in all four nebulizers simultaneously. Increasing the number of inhaled 99mTc-DTPA particles caused an increase in PCl of 24.2% (P less than 0.01). We conclude that there is a slight but significant effect of changing the number of DTPA particles on PCl and that this is probably due to an uneven mixing of the 99mTc-DTPA in the aqueous hypophase underlying the surfactant lining and the alveoli.