Comparison of ccd of F, parDE of RP4, and parD of R1 using a novel conditional replication control system of plasmid R1

A number of plasmid-encoded gene systems are thought to stabilize plasmids by killing plasmid-free cells (also termed post-segregational killing or plasmid addiction). Here we analyse the mechanisms of plasmid stabilization by ccd of F, parDE of RP4 and parD of R1, and compare them to hok/sok of R1. To induce synchronous plasmid loss we constructed a novel plasmid replication-arrest system, which possesses the advantage that plasmid replication can be completely arrested by the addition of IPTG, a non-metabolizable inducer. Using isogenic plasmid constructions we have found, for the first time, consistent correlation between the effect on steady-state loss rates and the effect on cell proliferation in the plasmid replication-arrest assay for all three systems. The parDE system had the most pronounced effect both on plasmid stabilization and on plasmid retention after replication arrest. In contrast, ccd and parD both exhibited weaker effects than anticipated from previously published results. Thus, our results indicate that the function and efficiencies of some of the systems should be reconsidered. Our results are consistent with the previously postulated hypothesis that ccd and parDE act by killing plasmid-free segregants, whereas parD seems to act by inhibiting cell division of plasmid-free segregants.     

Is hydrogen peroxide a second messenger of salicylic acid in systemic acquired resistance?

Elevated levels of salicylic acid (SA) are required for the induction of systemic acquired resistance (SAR) in plants. Recently, a salicylic acid-binding protein (SABP) isolated from tobacco was shown to have catalase activity. Based on this finding elevated levels of hydrogen peroxide (H₂O₂) were postulated to act as a second messenger of SA in the SAR signal transduction pathway. A series of experiments have been carried out to clarify the role of H₂O₂ in SAR-signaling. No increase of H₂O₂ was found during the onset of SAR. Induction of the SAR gene, PR-1, by H₂O₂ and H₂O₂-inducing chemicals is strongly suppressed in transgenic tobacco plants that express the bacterial salicylate hydroxylase gene, indicating that H₂O₂ induction of SAR genes is dependent on SA accumulation. Following treatment of plants with increasing concentrations of H₂O₂, a dose-dependent accumulation of total SA species was found, suggesting that H₂O₂ may induce PR-1 gene expression through SA accumulation. While the results do not support a role for H₂O₂ in SAR signaling, it is suggested that SA inhibition of catalase activity may be important in tissues undergoing a hypersensitive response.     

Localization of visual pigments within rhabdoms of the compound eye of Spodoptera exempta (Insecta,Noctuidae)

Summary In the noctuid moth Spodoptera exempta, the distribution of visual pigments within the fused rhabdoms of the compound eyes was investigated by electron microscopy. Each ommatidium regularly contains eight receptor cells belonging to three morphological types: one distal, six medial, and one basal cell (Meinecke 1981); four different visual pigments — absorption maxima at approximately 355, 465, 515, and 560 nm — are known to occur within the eye (Langer et al. 1979). The compound eyes were illuminated in situ by use of monochromatic light of different wavelengths. This illumination produced a wide scale of structural changes in the microvilli of the rhabdomeres of individual cells. Preparation of eyes by freeze-substitution revealed the structural changes in the rhabdomeres to be effects of light occurring in vivo.The degree of structural changes may be considerably different in rhabdomeres within the same ommatidium; it was found to depend on the wavelength and the duration of illumination, the intensity received by the ommatidia as well as the spectral sensitivity of the receptor cells. Therefore, it was possible to estimate the spectral sensitivities of the morphological types of receptor cells. Generally, all medial cells are green receptors and all basal cells red receptors; distal cells are blue receptors in about two-thirds of the ommatidia, while in the remaining third of them distal cells are sensitive to ultraviolet light.Supported by Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 114 (Bionach)     

Repair rate in human fibroblasts measured by thymine dimer excorporation

Summary The UV photoproduct, thymine dimer ( ), is excorporated with a remarkably low rate from the DNA of human fibroblasts grown in cell culture. An UV dose of 18 J/m² creates 0.045% (related to thymine). Within the first two days of repair logarithmically growing and quiescent fibroblasts exhibit the same repair rates; thereafter, the proportion of is lower in growing cells due to recovery of DNA replication. Only about 50% of the lesions are excised within 24 h. In quiescent cells, 13% of the thymine dimers originally present can be detected as late as a week after UV-irradiation. Two distinct first-order rate constants indicate that approximately half of the dimers are less accessible to repair. Repair measured by the nucleoid decondensation technique corresponds to the faster repair rate, whereas the slow repair rate cannot be detected by this method. Saturation of repair is found beyond 27 J/m². The remarkably slow rate of excision indicates that thymine dimers are not lethal lesions in human fibroblasts.     

Hydrophobic interaction of soluble 3′:5′ cyclic nucleotide phosphodiesterase with octyl-sepharose

Summary Soluble cyclic nucleotide 3:5 monophosphate phosphodiesterase (PDE) (EC 3.1.4.17) obtained from beef adrenal cortex as the 100,000 g/1.5 h supernatant is usually regarded as a very hydrophilic protein. However, when subjected to hydrophobic chromatography on Octyl-Sepharose CL 413 it reveals strong hydrophobic interaction with the column matrix. The chromatographic procedure leads to multiple but distinct forms of PDE which degrade cAMP beyond 5AMP to inosine, via adenosine. The same metabolic pathway was previously observed with a membrane bound multienzyme sequence. Even the soluble PDE forms separated by gel chromatography (Sephadex G 200, Sepharose S 200 and Sepharose 6B) and soluble PDE of other tissue (heart) displayed the same metabolic pattern. These findings indicate a linkage between PDE, nucleotidase and deaminase activities. The intimate association of the enzyme is additionally supported by the phenomenon of kinetic advantage clearly observed with the most hydrophobic PDE form. Its end product, inosine, is formed more rapidly from CAMP than from the intermediate 5AMP. This paradoxical phenomenon is explained by close physical proximity between the enzymes involved in the metabolic pathway. Furthermore, when the most hydrophobic PDE form was immobilized on Octyl-Sepharose, rather than loss of catalytic activity even higher enzyme activities were measured. It is suggested that the so-called multiple forms of soluble PDE-at least in part-represent more or less preserved forms of a native, membrane bound, multienzyme sequence which degrades cyclic nucleotides.     

Nuclear cytochrome-deficient mutants of Neurospora crassa: isolation, characterization, and genetic mapping.

Summary We have isolated twenty-six nuclear, singlegene cytochrome-deficient mutants of Neurospora crassa as an initial step toward the study of the structural components and regulatory mechanisms involved in the biogenesis of the mitochondrial cytochrome system. These mutants, together with two previously described mutants, cyt-1 and cyt-2, have been classified into six distinct groups on the basis of cytochrome phenotype: a) cytochrome aa ₃ deficiency (due to mutations affecting loci designated cya); b) cytochrome b deficiency (cyb-1 locus); c) cytochrome b deficiency with a partial deficiency of cytochrome aa ₃ (cyb-2 locus); d) deficiency of both cytochromes aa ₃ and b (cyt loci); e) deficiency of both cytochromes aa ₃ and c (cyt-2 locus); and f) partial deficiency of cytochromes aa ₃ and c (cyt-12 locus).Four of seven mutations affecting cya loci have been mapped and are located on linkage groups I, II, V, and VI. It is not yet known whether these genes code for structural components of cytochrome oxidase or have a regulatory function that affects synthesis or assembly of the enzyme. The cyb-1 and cyb-2 genes are located on linkage groups V and VI, respectively, and appear to code for regulatory elements that control the biogenesis of cytochromes b and aa ₃ . The positions of the cyt mutations that cause a simultaneous deficiency of cytochromes aa ₃ and b are dispersed throughout the genome, except for two gene clusters on the left arm of linkage group I. Some of these mutants may be deficient in mitochondrial protein synthesis. Two mutations, cyt-2 and cyt-12, are located on linkage groups VI and II, respectively, and appear to affect genes that code for components of a regulatory system that controls the biogenesis of cytochromes aa ₃ and c.     

Mass spectrometry of silylated flavonol O-glycosides

The electron impact mass spectra of 19 trimethyl silylated flavonol mono-, di- and -triglycosides are reported for the first time. All spectra show wel     

Ammoniteli der GattungParapatoceras aus dem Oberen Mitteljura des Süntels (östliches Wesergebirge,Niedersachsen)

The Parapatocerates from the Süntel (Weser mountains, Northwest Germany) are described being an utmost variable subspecies respectively geographical raceParapatoceras distans (?) bentzi (Potonié) with two possibly modificatorily separated form groups. They differ from the similarP. distans(?) crioconus (Buckman) (=Crioconites Buckman) of Chippenham (England) in primary quadrilobaty and further characteristics accessible only by precise analysis. To establish the speciesdistans Baugier & Sauzé and with this the genusParapatoceras, recovery of the holotype or a determination of a neotype is required. A very similar but probably quinquelobate specimen (Parapatoceras sp.) from the Macrocephalus oolite of Hanover seems to accentuate once more the taxonomically minor degree of the quadri- or quinquelobaty inParapatoceras.     

Temporal study of the staphylococci and micrococci of normal infant skin.

Staphylococcus and Micrococcus populations were collected from the healthy skin of 10 infant subjects. Infants were sampled from 1 day to 32 weeks of age. Species were characterized by approximately 30 different morphological, physiological, and biochemical characters. Staphylococci were the predominant inhabitants of normal skin, whereas micrococci were found only occasionally in this environment. Staphylococcus epidermidid, S. haemolyticus, and S. hominis were the predominant and persistent staphylococci. These species constituted a high percentage of the total aerobic bacterial flora of infant skin. Micrococcus luteus and M. kristinae were the prevalent micrococci found on infant skin. Only limited correlation between Staphyloccus and Micrococcus populations and infant age or body area sampled was indicated by this study.