Active transport of [32P]thiamine diphosphate inEscherichia coli

Summary Active uptake of [³²P]thiamine diphosphate byE. coli was analyzed using an improved method of gel filtration chromatography. The radioactive coenzyme was accumulated without dephosphorylation. From this result it was concluded that thiamine kinase is not involved in the membrane transport of thiamine inE. coli.We are indebted to Miss M. Abe for her technical assistance.     

Versuche zur Wirkungssteigerung der Kombination Sulfamethoxazol/Trimethoprim durch EDTA und Levallorphan anEscherichia coli

Summary EDTA, which is reported to potentiate the action of some antibiotics and chemotherapeutics in various bacterial strains, showed only additive effects inE. coli, when combined with trimethoprim or sulfamethoxazole, eithersingly or together. Neither was a potentiation seen with sulfanilamide, the permeation and effectiveness of which is less pronounced than that of sulfamethoxazole. The same negative results were obtained with levallorphan, which leads to alterations in the cell membrane. EDTA may therefore be of chemotherapeutic interest only in special cases, e.g. inPsuedomonas species which possess a highly EDTA-sensitive cell wall.     

Mg2+-ATPase defective mutant ofEscherichia coli and thiamine transport

Summary Mg²⁺-ATPase deficient mutant ofEscherichia coli showed an evident dependency of thiamine uptake on the oxidative metabolism of glucose, whereas the parent strain did not. In both cells, this uptake was completely inhibited by H⁺ conductors.Acknowledgment. We are indebted to Miss M. Abe for her excellent technical assistance.     

The effect of azetidine-2-carboxylic acid on the synthesis of proline inEscherichia coli

Zusammenfassung Die Beeinflussung der Prolinsynthese inEscherichia coli durch Azetidin-2-carbonsäure wird untersucht.

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This report is part of a thesis submitted byFaith I. Smith to the Graduate School at Oregon State University in partial fulfillment of the requirements for a M.S. degree.     

Inhibition of the infective activity of phage f2 and its infectious RNA by histone

Zusammenfassung Histon hemmt die Entwicklung der Phagen f₂ inE. coli-K 13-Zellen und in den Spheroplasten, die mit f₂ RNS infiziert wurden. Die inaktivierende Wirkung des Histons auf freie Phagenpartikel und infektiöse RNS wurde beschrieben.     

Histidine-induced injury to cultured liver cells,effects of histidine derivatives and of iron chelators

The amino acid histidine is an excellent buffer and is therefore included in several organ preservation solutions used in transplantation medicine. However, when used at concentrations as in these solutions, histidine has a marked injurious potential. Therefore, we here assessed the mechanism of histidine-induced cell injury and searched for ways to use the buffering power of histidine but avoid histidine toxicity. When cultured hepatocytes were incubated in HTK solution or in modified Krebs-Henseleit buffer containing 198 mM L-histidine at 37°C, most cells lost viability within 3 h (LDH release 86 ± 7% and 89 ± 5%, respectively). This injury was accompanied by marked lipid peroxidation, and was strongly inhibited by hypoxia, by the antioxidants trolox, butylated hydroxytoluene and N-acetylcysteine and by the membrane-permeable iron chelators 2,2′-dipyridyl, 1,10-phenanthroline, LK 614, LK 616 and deferoxamine. Thus, histidine-induced cell injury appears to be mediated by an iron-dependent formation of reactive oxygen species. D-Histidine, imidazol and L-histidine methyl ester also elicited marked injury, while the N-substituted derivatives Nα-acetyl-L-histidine and tert-butyl-oxycarbonylhistidine and histidine-containing dipeptides showed almost no toxicity. Histidine toxicity, its iron dependence and the superiority of Nα-acetyl-L-histidine were also evident during/after cold (4°C) incubations. Therefore, we suggest the addition of iron chelators to histidine-containing solutions, and/or replacing histidine with Nα-acetyl-L-histidine in organ preservation solutions. Received 23 October 2006; accepted 21 November 2006     

Induction of bacteriophage and colicin by means of acridine orange

Zusammenfassung Es wird über Ergebnisse bei der Induktion der Phage- und Colicinsynthese inE. coli (Stamm 18) durch photodynamische Wirkung des Acridinorange berichtet. Der Phagentiter wurde 11mal, der Colicintiter kaum 2mal erhöht. Acridinorange zeigte ohne Belichtung keinen deutlichen Effekt.     

Selective inhibition of the multiplication of phage T1 inE. coli K12

Riassunto Un nuovo antibiotico, la Distamicina, inibisce selettivamente lo sviluppo del batteriofago T1 inE. coli K12 pur non esercitando alcuna azione sullo sviluppo del microorganismo ospite. L'antibiotico può inibire sia la penetrazione del batteriofago nelle cellule batteriche sia la sua liberazione.     

A study of granulated metrial gland cell differentiation in pregnant,macrophage-deficient,osteopetrotic (op/op) mice

A population of uterine natural killer (NK) cells, commonly called granulated metrial gland (GMG) cells, differentiates in the mouse uterus during normal pregnancy. Little is known regarding the process of differentiation of GMG cells or of other NK cell subsets. It has been suggested that macrophage precursors, under the combined influences of the cytokine growth factors colony stimulating factor-1 (CSF-1) and interleukin-2, become NK-cell like in morphology, pattern of target cell lysis and surface antigen phenotype. Mice expressing the mutation osteopetrosis (op/op) are unable to produce the cytokine CSF-1. To determine whether CSF-1 is required for the successful differentiation of uterine NK cells, implantation sites in pregnant,op/op mice were studied histologically. GMG cell differentiation appeared to progress normally inop/op mice studied between days 7 and 14 of gestation. Thus, the growth factor CSF-1 is not required for differentiation of the uterine NK cell subset known as GMG cells and probably GMG cells do not differentiate from macrophage precursor cells which are deficient inop/op mice.     

Periplasmic lysozyme inhibitor contributes to lysozyme resistance in <Emphasis Type="Italic">Escherichia coli</Emphasis>

The product of the Escherichia coli ORFan gene ykfE was recently shown to be a strong inhibitor of C-type lysozyme in vitro. The gene was correspondingly renamed ivy (inhibitor of vertebrate lysozyme), but its biological function in E. coli remains unknown. In this work, we investigated the role of Ivy in the resistance of E. coli to the bactericidal effect of lysozyme in the presence of outer-membrane-permeabilizing treatments. Both in the presence of lactoferrin (3.0 mg/ml) and under high hydrostatic pressure (250 MPa), the lysozyme resistance of E. coli MG1655 was decreased by knock-out of Ivy, and increased by overexpression of Ivy. However, knock-out of Ivy did not increase the lysozyme sensitivity of an E. coli MG1655 mutant previously described to be resistant to lysozyme under high pressure. These results indicate that Ivy is one of several factors that affect lysozyme resistance in E. coli, and suggest a possible function for Ivy as a host interaction factor in commensal and pathogenic E. coli.Received 12 February 2004; received after revision 11 March 2004; accepted 16 March 2004     

Blockade by zinc ions of K+-induced contraction and calcium in guinea pigTaenia coli

Preincubation with 0.3 mM Zn²⁺ markedly inhibited both the tonic response and Ca²⁺ binding at low affinity sites induced by K⁺ (60 mM), with smaller effects on the phasic response and the high affinity Ca²⁺ sites, inTaenia coli. However, when the muscle was kept in Zn²⁺-containing medium following the first stimulation with the K⁺, the phasic response and the high affinity Ca²⁺ sites were more severely inhibited during the second stimulation with K⁺. This probably indicates that Zn²⁺ reduced the tonic tension response to K⁺ mainly by inhibiting Ca²⁺ influx at the cell membranes ofTaenia coli. However, when Zn²⁺ is continuously present, Ca²⁺ is not supplied at the storage sites and is not available for the phasic response to a second stimulation with K⁺.     

The karyotype,C-bands and AgNO3-bands of a lungless salamander from Korea:Onychodactylus fischeri (Boulenger) (Amphibia,Urodela)

The karyotype of a lungless salamander,Onychodactylus fischeri, from Korea was analyzed and compared with that of the Japanese congeneric species,O. japonicus. In both species the diploid karyotype consists of78 chromosomes, including 6 pairs of large chromosomes, 6 pairs of medium-sized ones, and the remaining 27 pairs of microchromosomes. The chromosome number ofO. fischeri, 2n=78, is, like that ofO. japonicus, the largest so far reported in the order Urodela. C-banding showed that constitutive heterochromatin inO. fischeri was mainly in the centromeric regions and near the secondary constrictions of the large chromosomes. AgNO₃-bands were located in the secondary constrictions associated with C-band heterochromatin.     

Splenic B-cell activation in lipopolysaccharide-non-responsive C3H/HeJ mice by lipopolysaccharide ofPorphyromonas gingivalis

Porphyromonas gingivalis 381 lipopolysaccharide (LPS) definitely exhibited mitogenic activity in purified B-cells, separated from spleens of LPS-responsive C3H/HeN mice and LPS-non-responsive C3H/HeJ mice by using a magnetic cell sorting system. The mitogenic activity induced byP. gingivalis LPS was incompletely inhibited by polymyxin B.P. gingivalis LPS also induced a higher production of interleukin-6 (IL-6) in splenic B-cells of C3H/HeN mice as compared withEscherichia coli LPS. Furthermore,P. gingivalis LPS, but notE. coli LPS, induced definite IL-6 production in C3H/HeJ mice.P. gingivalis LPS increased tyrosine, serine/threonine phosphorylation of proteins with various major induced bands in splenic B-cells of both C3H/HeN and C3H/HeJ mice. Additionally, radioiodinatedP. gingivalis LPS, similarly toE. coli LPS, bound to a 73-kDa protein on C3H/HeJ as well as C3H/HeN B-cells. ThusP. gingivalis LPS may activate B-cells of C3H/HeJ as well as C3H/HeN mice via the LPS-specific binding protein on the cells.     

Selective blockade of components of potassium activation inMyxicola axons

Summary The K⁺ conductance inMyxicola giant axons activates in two phases which are pharmacologically separable. The fast phase of K⁺ activation is specifically inhibited by 4-aminopyridine and by the substitution of D₂O for H₂O. We suggestMyxicola giant axons, like the amphibian node of Ranvier, may possess more than one variety of K⁺ channel.     

Acetic acid vapour as a resource: Threshold differences among threeDrosophila species

Summary Acetic acid vapour is utilized as an energy source up to a threshold where it becomes a stress in 3Drosophila species. The threshold ranking isD. melanogaster» D. simulans>D. immigrans, which qualitatively parallels that for ethanol. The capacity to use nutritive vapours appears to be very important inDrosophila ecology.I thank Gary Spence for technical assistance, and the Australian Research Grants Committee for partial financial support.     

On the association between lysogeny and carcinogenicity in nitroquinolines and related compounds

Zusammenfassung In einer Serie von 16 Nitroquinolinen und verwandten Substanzen wurde eine gute Übereinstimmung zwischen Phageninduktion inEscherichia coli C-600 und Karzinogenese nachgewiesen.

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Supported by grants No. C-6516 and FR-05526 from the National Cancer Institute and Contract NO. PH 86-66-169 from the Division of Air Pollution, U.S.P.H.S. We thank Dr.Y. Kawazoe of the National Cancer Center Research Institute, Tokyo, for his generous gift of the compounds tested.     

Activation concomitante par l'hétéroauxine de la résorption du corps paranucléaire ribonucléique et de la germination des zygotes chezAllomyces

Summary Heteroauxin (optimal concentration 10^–5) activates the germination of the zygotes and the dissolution of their RNA nuclear cap inAllomyces javanicus.Alkaline phosphatase has been located cytochemically in the cytoplasm surrounding the RNA nuclear cap.These new observations lead the author to postulate a relationship between the stimulating action of heteroauxin, activation of the phosphatases and awakening of the RNA-protein synthesis dynamic system during the germination of the zygotes inAllomyces.     

Genetic analysis of modifier variability inDrosophila subobscura

Summary The previously detected modifier variability acting on the expression of the Bare (Ba) locus inDrosophila subobscura is demonstrated to be due to polygenes situated along the chromosome O. From crosses between isogenic lines of high and low modifier effect we ascertained the presence of approximately 5 modifier loci.We are grateful to Dr M. Green for valuable comments on the experimental design used in this paper.     

Effect of xanthurenic acid on P-450-dependent biotransformation by molting glands in vitro

Incubation of molting glands from the crayfishProcambarus clarkii (Y-organ) and the silkwormBombyx mori (prothoracic gland) with 23,24-[²H₄]-2-deoxyecdysone resulted in the production of deutero-ecdysone; this biotransformation was inhibited in the presence of xanthurenic acid. When the experiments were performed under an¹⁸O₂ atmosphere, the¹⁸O atom was introduced into ecdysone, as confirmed by mass spectrometry. We therefore suggest that xanthurenic acid inhibits P-450-dependent hydroxylation of 2-deoxyecdysone. However, deutero-2-deoxyecdysone was not converted to 3-dehydroecdysone when using Y-organs in vitro, although it is a major product. We therefore conclude that the biosynthetic pathway of ecdysteroids inP. clarkii branches at an early step.     

Presence of benzodiazepine binding sites (receptors) and amplification thereof by imprinting inTetrahymena

Summary LiveTetrahymena cells bound³H-diazepam specifically, as demonstrated by autoradiographic evidence of displacement of about 25% of labeled diazepam in the presence of a 1000-fold amount of cold diazepam. The³H-diazepam bound to membrane preparations isolated from untreated (control) cells was not displaced by cold diazepam, whereas cells involved in primary interaction (imprinting) with diazepam showed amplification and specificity of diazepam binding in both in vivo (cell suspension) and in vitro (pellicle) systems, as well as displacement of bound label in the presence of 1000-fold cold diazepam. It appears that diazepam induced imprinting and, consequently, also the formation of specific receptors inTetrahymena.