Characterization of the ugp region containing the genes for the phoB dependent sn-glycerol-3-phosphate transport system of Escherichia coli

Summary The ugp structural genes, coding for the pho regulon dependent sn-glycerol-3-phosphate transport system, were cloned in pBR322 and characterized. The expression of the cloned ugp system was phoB dependent. Cells containing the ugp plasmid overproduced the G3P binding protein upon phosphate starvation. Tn5 mutagenesis of the cloned DNA revealed that the ugp genes are organized in two separate operons which comprise at least four genes: ugpB and ugpD constitute one operon, ugpA and ugpC constitute the other. The structural gene for the G3P binding protein (G3PBP) is ugpB.The ugpC gene product was also synthesized in minicells as a polypeptide, with an apparent molecular weight of 40,000. No gene products could be assigned to the ugpA and ugpD genes. Hybridization experiments allowed the physical characterization of 20 kb of DNA adjacent to the ugp genes on the E. coli chromosome including the liv genes.     

Trypanosome hybrids generated in tsetse flies by nuclear fusion.

Genetic exchange may occur between two particular Trypanosoma brucei clones simultaneously transmitted by the same tsetse fly. We report here that this exchange takes place in the fly, through nuclear fusion. The resulting hybrids appear to be sub-tetraploid, some particular DNA sequences from one of the parental stocks being lost before enough cloned hybrid trypanosomes could be harvested for DNA analysis. A further reduction of the DNA content of these hybrids occurs gradually upon growth and yields near diploid value in a major part of the population. This mode of hybrid generation is different from the fusion of haploid gametes, which is thought to occur normally upon inoculation of metacyclic trypanosomes in their mammalian host. In this respect, the sub-tetraploid hybrids appear to undergo meiosis in the fly, generating sub-diploid metacyclic forms, then fusion in the mammalian blood.     

Control of echolocation pulses by neurons of the nucleus ambiguus in the rufous horseshoe bat,Rhinolophus rouxi

Horseradish peroxidase was applied by inotophoretic injections to physiologically identified regions of the laryngeal motor nucleus, the nucleus ambiguus in the CF/FM bat Rhinolophus rouxi. The connections of the nucleus ambiguus were analysed with regards to their possible functional significance in the vocal control system, in the respiration control system, and in mediating information from the central auditory system. The nucleus ambiguus is reciprocally interconnected with nuclei involved in the generation of the vocal motor pattern, i.e., the homonomous contralateral nucleus and the area of the lateral reticular formation. Similarly, reciprocal connections are found with the nuclei controlling the rhythm of respiration, i.e., medial parts of the medulla oblongata and the parabrachial nuclei. Afferents to the nucleus ambiguus derive from nuclei of the 'descending vocalization system' (periaqueductal gray and cuneiform nuclei) and from motor control centers (red nucleus and frontal cortex). Afferents to the nucleus ambiguus, possibly mediating auditory influence to the motor control of vocalization, come from the superior colliculus and from the pontine nuclei. The efferents from the pontine nuclei are restricted to rostral parts of the nucleus ambiguus, which hosts the motoneurons of the cricothyroid muscle controlling the call frequency.     

Diadenosine 5′,5‴-P1,P3-triphosphate in eukaryotic cells: Identification and quantitation

A highly sensitive enzymatic assay for diadenosine 5′,5?-P¹,P³-triphosphate (Ap₃A) has been established on the basis of the coupled luminescence assay for diadenosine 5′,5?-P¹,P³-tetraphosphate (A. Ogilvie (1981)Anal. Biochem.115, 302–307). Snake venom phosphodiesterase splits Ap₃A into AMP plus ADP which can be measured in a luminescence reaction containing pyruvate kinase, phosphoenolpyruvate and luciferin-luciferase. The procedure is linear with Ap₃A levels ranging from 0.1 to 2 pmol. The assay has been used to measure Ap₃A in various eukaryotic cells after ion-exchange chromatography and high-performance liquid chromatography of acidic extracts of the cells. The level of diadenosine triphosphate was higher in all instances than the level of diadenosine tetraphosphate. When growing in the abdominal cavity of mice, Ehrlich ascites tumor cells contained high amounts of Ap₃A ($\text{0.1}\text{nmol}\text{10}{}^6\text{cells}$), allowing direct optical determination in the HPLC chromatography. The quantitative measurement of Ap₃A with the luminescence assay gave identical results. Ap₃A extracted from Ehrlich cells was also chromatographed with authentic nucleotide in two thin-layer systems providing additional proof for the existence of Ap₃A in biological material.     

Desmethylimipramine Enhances the Release of Endogenous GABA and Other Neuro trans mitter Amino Acids from the Rat Thalamus

The influence of desmethylimipramine (DMI) on the release of endogenous gamma-aminobutyric acid (GABA) and some other amino acids from the rat thalamus was studied with a push-pull perfusion technique. Following HPLC the amino acids were fluorimetrically estimated. Added to the perfusion medium at a concentration of 10 mumol L-1, DMI caused a 5- to 10-fold increase in the release of GABA. Similar effects were found with imipramine, trimeprimine, haloperidol, and propranolol. The elevation of GABA release induced by DMI was Ca dependent. The release of aspartate and glutamate was also increased by DMI, but in contrast to K ions, DMI did not reduce the thalamic output of glutamine.     

Saturated Fatty Acid Mutant of Saccharomyces cerevisiae with an Intact Fatty Acid Synthetase

To determine directly the effects of streptomycin on translational fidelity in intact cells, we studied the synthesis of beta-galactosidase and of the coat protein of bacteriophage R17 in an Escherichia coli mutant in which the bactericidal effects of streptomycin are delayed. After the addition of streptomycin to exponentially growing mutant cells, protein synthesis continues at an undiminished rate for approximately an hour; however, as measured by enzyme assays, little functional protein is produced. Serological assays designed to detect beta-galactosidase and bacteriophage R17 coat protein show that substantial amounts of the protein synthesized can react with antisera prepared against active beta-galactosidase and phage R17, indicating the aberrance of the protein produced in the presence of the antibiotic. The polypeptides synthesized in the presence of streptomycin are degraded in the cell to a much greater extent than protein synthesized in the absence of the antibiotic. The proteolytic attack on this protein is not affected by inhibitors of serine proteases, suggesting that enzymes other than those involved in "normal turnover" of cellular protein are responsible. In this strain, certain of the multiple effects of streptomycin are separated in time and the production of abnormal protein (enzymatically inactive and susceptible to proteolytic attack) could be studied in the absence of the lethal effect of the drug.     

Differential staining of plant chromosomes with Giemsa

Simple Giemsa staining techniques for revealing banding patterns in somatic chromosomes of plants are described. The value of the methods in the recognition of heterochromatin was demonstrated using five monocotyledonous and two dicotyledonous species. In Trillium grandiflorum the stronger Giemsa stained chromosome segments were shown to be identical with the heterochromatic regions (H-segments) revealed by cold treatment. Preferential staining of H-segments was also observed in chromosomes from three species of Fritillaria and in Scilla sibirica. Under suitable conditions the chromosomes of Vicia faba displayed a characteristic banding pattern and the bands were identified as heterochromatin. The Giemsa techniques proved to be more sensitive than Quinacrine fluorescence in revealing a longitudinal differentiation of the chromosomes of Crepis capillaris, where plants with and without B-chromosomes were examined. Again all chromosome types had their characteristic bands but there was no difference in Giemsa staining properties between the B-chromosomes and those of the standard complement.     

Patterns in the release of gaseous ammonia by terrestrial isopods

Summary In the fall and in early spring P. scaber and O. asellus released gaseous ammonia in the form of more or less regularly spaced bursts. In the spring about twice as much ammonia was released by O. asellus than in the fall. In late spring and summer, however, both species released ammonia in a rhythmic fashion, with a maximum at noon and early in the afternoon, and a minimum early at night. Sometimes a second maximum occurred late at night.In O. asellus the addition of a moist substrate to the reaction chamber shifted the maximum of the release of ammonia from noon to late night and early morning.Fed specimens of P. scaber released only about one-third as much NH₃ as fasting animals and—at least in constant darkness—with a period of much reduced amplitude.It is concluded that the rhythmical release of ammonia is inversely related to the pattern of locomotory activity of these animals. This would implicate mechanisms that regulate either the production or the release of ammonia in such a way that the maximum occurs at a time when the animals' production of energy is at a minimum and when they are protected against loss of water by sitting in their moist retreats.The work at Innsbruck was supported by the Fonds zur Förderung der wissenschaftlichen Forschung of Austria.     

The complete heart-lead relationship in the einthoven triangle

The relationship between the source strength and the “manifest vector” in the Einthoven Triangle is derived for a line and a point dipole source and confirmed experimentally. The result permits the interpretation of the standard ECG leads in absolute terms and corrected for body size. The manifest vector is shown to be approximately times what it would be in an otherwise similar circular slab which circumscribes the triangle.