Biosurfactant yields and nutrient consumption of Pseudomonas fluorescens 378 studied in a microcomputer controlled multifermentation system

Production of biosurfactant AP-6 and consumption of carbon (succinic acid) and nitrogen (ammonium ions) by Pseudomonas fluorescens 378 were studied under different growth conditions. The study was performed in a microcomputer controlled multibatch fermentation system which enabled simultaneous running of 10 fermentors. The fermentors were mantled glass vessels, temperature controlled by circulated water, and mixing was arranged by magnetic stirrers. They were connected to the computer system (pH measurement and control) via signal conditioning cards. The microcomputer had a 128 kbytes RAM, two 800-kbyte floppy disc drives, a graphic terminal, and expansion cards. Biosurfactant production was independent of the carbon-to-nitrogen ratio and the phosphorus content in the medium. Omitting the Fe(III) supplement to the medium increased the product yield by 120%. Changes in oxygen transfer rate and pH in the iron deficient cultures did not have any effect on the product yield. Iron deficiency increased the cell consumption of carbon source. Consumption of carbon source in relation to nitrogen uptake (carbon/nitrogen quotient) increased with increasing quotient in the growth medium. The uptake of carbon and nitrogen changed in the intervals of 1.2-1.5 g/g biomass and 0.09-0.16 g/g biomass, respectively. The consumption of carbon increased from 1.5 g/g biomass to 2.0 g/g biomass when the medium concentration of phosphorus was decreased from 0.18 to 0.027 g/L.     

Variability of measurements of cranial growth in the rabbit

This study concerns techniques used in experimental cranial growth research: roentgen cephalometry, roentgen stereophotogrammetry, and gross measurements (osteometry). A comparison of the precision of these methods has not been found in the literature. Computation of technical errors is fundamental to the sound evaluation of registered findings, and such a presentation must be obligatory in all biometric reports. We compared the measurement error of roentgen cephalometric and osteometric data with that obtained by roentgen stereophotogrammetry (RSA). RSA demonstrates a superior replicability, and this technique gives possibilities for kinematic and volumetric determinations simultaneously with distance evaluation. Roentgen cephalometry has the advantage of enabling distance and angular measurements between any well-defined skeletal points or lines. This technique, preferably after implantation of bone markers, is a reliable alternative, but optimal results necessitates calculations of the magnification factor for each bone segment involved. Direct osteometry does not contribute additional information, but problems of image magnification are omitted. Preferably, one individual should perform all measurements regardless of the method used. Growth rates and values calculated by one technique cannot be directly transformed to some other approach. In all probability, assessments of distance changes would gain substantially by using one technical approach consistently throughout actual age intervals. The least variable measurements of sutural growth are made for sutures growing primarily in one plane and with substantial growth rates. One must realize that differences among studies may be due to the limitations of, in particular, the cephalometric and osteometric techniques.     

Gene and mRNA for precursor polypeptide VI from adenovirus type 2.

We present a 1,040-base-pair-long sequences of adenoviruses type 2 DNA which encodes the complete gene for precursor polypeptide VI (pVI). pVI consists of 250 amino acids amounting to a molecular weight of 26,990. The proteolytic cleavage maturing pVI to virion polypeptide VI removes 33 amino acids from the amino-terminal end of the polypeptide, thus giving the mature polypeptide VI a molecular weight of 23,400. The UAA stop codon terminating pVI translation is separated by 84 nucleotides from the initiator triplet for the hexon gene. Both polypeptides are encoded by the same translational reading frame, suggesting the evolution of pVI and hexon as separate proteins by the introduction of a termination codon and selection of a new splice acceptor site in an ancestral fused polypeptide chain. The splice site where the common tripartite leader is attached to the pVI mRNA precedes the initiator codon for pVI translation by one nucleotide and forms, together with other late splice acceptor sites, a late adenovirus consensus acceptor site. We also demonstrate that the 3' end of the mRNA's belonging to the L2 3'-cotermination family is located only 31 nucleotides upstream from the splice junction of the pVI mRNA. Furthermore, we show that four novel polypeptides of molecular weights 80,000, 39,000, 36,000, and 10,500 are encoded by region L2.     

Virus-receptor interaction in the adenovirus system I. Identification of virion attachment proteins of the HeLa cell plasma membrane.

Plasma membranes from HeLa cells were isolated in a two-phase polymer system. To compare the efficiency of attachment protein extraction, a normalized assay for the assessment of adenovirus type 2 (Ad2) receptor-active components interfering with the attachment of Ad2 to HeLa cells was developed. An optimized detergent extraction procedure, 0.5% Triton X-100, was used, and solubilized membrane proteins were radioisotope labeled in vitro. Proteins with affinity for Ad2 virions were quantified and identified in a sucrose gradient sedimentation assay and by affinity chromatography with cross-linked Ad2 virions immobilized to AH-Sepharose 4B. From virions recovered in the sucrose gradient system, one major membrane component of high affinity was identified with a polypeptide molecular weight of around 40,000. Glycosylated proteins isolated by wheat germ lectin chromatography with high affinity for immobilized virus particles were isolated, and two major components with apparent molecular weights of 40,000 and 42,000 were identified. We suggest that a glycosylated protein with high affinity for Ad2 virions and a polypeptide molecular weight of 40,000 to 42,000 is one component of the Ad2 attachment site on HeLa cells.     

Control of Plasmid R1 Replication: Kinetics of Replication in Shifts Between Different Copy Number Levels

Plasmid R1 replication was studied in shifts between two steady states of copy number. The copy number was varied in two ways. First, we utilized the fact that it decreases with increasing growth rate. To minimize the metabolic effects of changes in the growth rate, the downshifts were obtained by adding α-methylglucoside to cultures growing in glucose-minimal medium, and the upshifts were obtained by adding glucose to cultures growing in the presence of glucose plus α-methylglucoside. Second, we used a temperature-dependent copy mutant of plasmid R1 (pKN301). Plasmid pPK301 shows a threefold higher copy number at 40 than at 30°C. In both types of shift, plasmid replication immediately adjusted to the postshift differential rate. The copy number asymptotically adjusted to the new steady state. Hence, the system that controls plasmid R1 replication sets the frequency of replication without measuring the actual copy number. It has been suggested that plasmid R1 replication is under negative control by an R1-mediated repressor protein. Among the replication control models that involve negative control, the Pritchard inhibitor dilution model, the Sompayrac-Maaløe autorepressor model, and the plasmid λdv system all predict gene dose-independent copy number control.     

Plasma Cortisol in the Horse,Diurnal Rhythm and Effects of Exogenous Acth

Peripheral blood plasma Cortisol concentration and its diurnal variation was measured in 4 horses. Mean concentration of Cortisol during 24 hrs. was 42 ng/ml (s ± 20 ng/ml). Peak values occurred at 6 a.m. and the lowest values were observed at about 6 p.m. (mean 65 ng/ml and 20 ng/ml, respectively). Long-acting ACTH at a dose of 150 i.u. was given by intramuscular injection to the 4 horses. Peak Cortisol concentrations markedly exceeding the prestimulation level were obtained between 2 and 4 hrs. after injection. During the immediate 24 hrs. after these peaks, the mean Cortisol level was markedly lower and the cyclic variation out of phase with the basal diurnal pattern. After a gradual adjustment during the second postinjection day, no differences could be seen between the 2 patterns on day 3 after injection.     

Allelic Negative Complementation at the Abruptex Locus of DROSOPHILA MELANOGASTER

The mutations of the Abruptex locus in Drosophila melanogaster fall into three categories. There are recessive lethal alleles and viable alleles. The latter can be divided into suppressors and nonsuppressors of Notch mutations. The recessive lethals are lethal in heterozygous combination with Notch. As a rule the recessive lethals are lethal also in heterozygous combination with the viable alleles. Heterozygous combinations of certain viable alleles are also lethal. In such heterozygotes, one heteroallele is a suppressor of Notch and the other is a nonsuppressor. Other heterozygous combinations of viable alleles are viable and have an Abruptex phenotype. The insertion of the wild allele of the Abruptex locus as an extra dose (carried by a duplication) into the chromosomal complement of the fly fully restores the viability of the otherwise lethal heterozygotes if two viable alleles are involved. The extra wild allele also restores the viability of heterozygotes in which a lethal and a suppressor allele are present. If, however, a lethal and a nonsuppressor are involved, the wild allele only partly restores the viability, and the effect of the wild allele is weakest if two lethal alleles are involved. It seems likely that of the viable alleles the suppressors of Notch are hypermorphic and the nonsuppressors are hypomorphic. The lethal alleles share properties of both types, and are possibly antimorphic mutations. It is suggested that the locus is responsible for a single function which, however, consists of two components. The hypermorphic mutations are defects of the one component and the hypomorphic mutations of the other. In heterozygotes their cumulative action leads to decreased viability. The lethal alleles are supposed to be defects of the function as a whole. The function controlled by the locus might be a regulative function.