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81.
A Mycobacterium leprae-specific gene encoding an immunologically recognized 45 kDa protein 总被引:1,自引:0,他引:1
Tobias F. Rinke de Wit Josephine E. Clark-Curtiss Feseha Abebe Arend H. J. Kolk Anneke A. M. Janson Miranda van Agterveld Jelle E. R. Thole 《Molecular microbiology》1993,10(4):829-838
By screening a Mycobacterium leprae lambda gt11 expression library with a serum from an Ethiopian lepromatous leprosy (LL) patient a clone was isolated (LL4) belonging to hybridization group III of a panel of previously isolated M. leprae clones. Members of this hybridization group encode a serologically recognized 45 kDa protein. The complete DNA sequences of the partially overlapping clones LL4 and L1 (hybridization group III) are presented and these revealed the presence of an open reading frame (ORF) predicting a protein with a molecular size of 42 448 Da. Southern hybridizations on total genomic DNA of M. Ieprae, Mycobacterium tuberculosis and eight atypical mycobacteria showed that the LL4 DNA fragment is specific for M. Ieprae DNA even under low-stringency conditions. The M. Ieprae specificity of LL4 DNA was further confirmed by the polymerase chain reaction using four different sets of primers. Western blotting analyses showed that the M. Ieprae 45 kDa protein is frequently recognized by antibodies from leprosy patients and that this recognition is specific since no antibodies could be detected in sera of tuberculosis patients. T-cell proliferation assays also demonstrated T-cell recognition by leprosy patients and healthy contacts of the M. Ieprae 45 kDa protein. The specificity of the LL4 DNA region and the 45 kDa antigen that is encoded by hybridization group III could provide unique tools for the development of M. Ieprae-specific immunological and DNA reagents. 相似文献
82.
R. W. H. M. van Tol J. H. Visser & M. W. Sabelis 《Entomologia Experimentalis et Applicata》2004,110(2):145-150
The vine weevil Otiorhynchus sulcatus is a parthenogenetic reproducing species which forages for suitable host plants at night, but is found congregated in dark places during the day. Frass of this weevil species is suspected to contain attractive compounds that are host‐plant related. Using a still‐air olfactometer, we tested adult vine weevils at night for their behavioural response to odours from conspecifics, feeding on a mixture of spindle tree (Euonymus fortunei) and yew (Taxus baccata), and to a sexually reproducing related species (Otiorhynchus salicicola), feeding on a mixture of ivy (Hedera helix) and cherry laurel (Prunus laurocerasus). Their attraction to conspecifics and O. salicicola appeared to be related to frass production. Freshly collected frass from O. sulcatus and from O. salicicola males and females was attractive. Prunus laurocerasus and H. helix have not been observed to be hosts of the vine weevil in the field. However, our tests showed that the vine weevil was attracted to mechanically damaged leaves of both plant species, whereas undamaged leaves were not attractive. Only undamaged young unfolding leaves of H. helix were also attractive. The attraction to odours from mechanically damaged host and non‐host plants suggested the involvement of compounds that are commonly found in many plant species. The involvement of plant compounds and/or aggregation pheromones in attraction to frass of the vine weevil and frass of the related weevil species O. salicicola is discussed. 相似文献
83.
Elise Flipse Marja G. M. Schippers Elly M. Janssen Evert Jacobsen Richard G. F. Visser 《Molecular breeding : new strategies in plant improvement》1996,2(3):211-218
Theamylose-free (amf) potato mutant can easily be complemented through introduction of the wild-type gene coding for granule-bound starch synthase (GBSS). After iodine staining the starch of theamf mutant is red whereas that of the wild type and the complementedamf mutant is blue. The level of complementation of selected transformants and their sexual off-spring after backcrossing withamf was investigated using sporophytic tuber cells and gametophytic microspore cells. Two diploid and two tetraploid transformants with full complementation demonstrated the expected segregation patterns of 1:1 (one active insert) or 3:1 (two independently segregating active inserts) in the microspores and in the F1 offspring based on staining of tubers. All expected genotypes in the F1 generation were found, based on microspore segregation patterns of the individual F1 plants. Two transformants with partial complementation (mixed phenotypes) were investigated. One of them, B1, was tetraploid and duplex for the GBSS insert, which had originated through mitotic doubling of the transformed diploid cells. In the F1 generation three phenotypic classes were found:amf, fully complemented and partially complemented. The latter two classes exist independently of a simplex or duplex gene status. The second transformant with partial complementation, B10, appeared to have a complex molecular composition. One cluster of five transgenes caused the partial complementation. Fully and partially complemented phenotypic classes were found after crossing B10 with theamf mutant. Indications were found that the ploidy level of the tissue in which the genes were introduced and expressed played an important role. Firstly, partial complementation was found after transformation of the diploid and not of the tetraploidamf genotypes. Secondly, the level of complementation was higher in tissue with lower ploidy levels, as illustrated by the colour of the starch inin vitro tubers (2x–4x cells) versus field-grown tubers (16x–64x). 相似文献
84.
Pieter H. B. De Visser Willem G. Keltjens Günther R. Findenegg 《Trees - Structure and Function》1996,10(5):301-307
In a pot trial growth and transpiration of 3-year-old Douglas-fir seedlings on an acid, sandy soil was examined at a deficient
(30 kg N ha –
1 year –
1) and an excessive level (120 kg N ha –
1 year –
1) of NH4 application. Dissolved ammonium sulphate was applied to the pots weekly for two growing seasons. In half of the pots a complete
set of other nutrients was applied in optimal proportions to the applied nitrogen. Water supply was optimal and transpiration
was recorded. At the end of the second treatment season irrigation was stopped for 2 weeks during dry and sunny weather. Both
high application of NH4 and additional nutrients increased shoot growth and transpiration demand in the first treatment year. The root system was
smaller at higher N level and this reduced water uptake accordingly. In the second year the combination of high NH4
+ and additional nutrients affected root functioning predominantly due to salinity effects and this seriously decreased water
uptake capacity and shoot water potentials, finally resulting in tree death. Without addition of other nutrients the high
NH4
+ application resulted in a high degree of soil acidification, which damaged the roots, that showed a decrease in water uptake
capacity. At the low NH4 supply level soil acidification was lower, and root functioning was not affected, and the trees recovered quickly from the
imposed drought. Higher needle K and P status depressed transpiration rates at the low NH4 application rate.
Received: 9 January 1995 / Accepted: 18 September 1995 相似文献
85.
Petra R. Moog Tom A. W. van der Kooij Wolfgang Brüggemann John W. Schiefelbein Pieter J. C. Kuiper 《Planta》1995,195(4):505-513
Arabidopsis thaliana (L.) Heynh. Columbia wild type and a root hair-less mutant RM57 were grown on iron-containing and iron-deficient nutrient solutions. In both genotypes, ferric chelate reductase (FCR) of intact roots was induced upon iron deficiency and followed a Michaelis-Menten kinetic with a K
m of 45 and 54 M FeIII-EDTA and a V
max of 42 and 33 nmol Fe2+·(g FW)–1·min–1 for the wild type and the mutant, respectively. The pH optimum for the reaction was around pH 5.5. The approximately four fold stimulation of FCR activity was independent of formation of root hairs and/or transfer cells induced by iron deficiency. Iron-deficiency-induced chlorosis and the development of a rigid root habit disappeared when ferric chelate was applied to the leaves, while FCR activity remained unchanged. The time course of the responses to iron deficiency showed that morphological and physiological responses were controlled separately.Abbreviations FCR
ferric chelate reductase
- FW
fresh weight
Thanks are due to Klaas Sjollema (Department of Electronmicroscopy, University of Groningen, The Netherlands) for help with the electron microscopy sample preparation and especially to Dr. Uwe Santore (Heinrich-Heine-University for electron microscopy. This work was supported by the SCIENCE programm of the European community; P.R.M.) and a Personal Research Grant by the Ministerium für Wissenschaft und Forschung of Nordrhein-Westfalen (P.R.M.) and last, not least by the productive discussions in ECOTRANS B.V. 相似文献
86.
Isolation and characterization of the mycobacterial phagosome: segregation from the endosomal/lysosomal pathway 总被引:5,自引:0,他引:5
Zahra Hasan Claudia Schlax Lotte Kuhn Ivan Lefkovits Douglas Young Jelle Thole & Jean Pieters 《Molecular microbiology》1997,24(3):545-553
Mycobacteria have the ability to persist within host phagocytes, and their success as intracellular pathogens is thought to be related to the ability to modify their intracellular environment. After entry into phagocytes, mycobacteria-containing phagosomes acquire markers for the endosomal pathway, but do not fuse with lysosomes. The molecular machinery that is involved in the entry and survival of mycobacteria in host cells is poorly characterized. Here we describe the use of organelle electrophoresis to study the uptake of Mycobacterium bovis bacille Calmette Guerin (BCG) into murine macrophages. We demonstrate that live, but not dead, mycobacteria occupy a phagosome that can be physically separated from endosomal/lysosomal compartments. Biochemical analysis of purified mycobacterial phagosomes revealed the absence of endosomal/lysosomal markers LAMP-1 and β-hexosaminidase. Combining subcellular fractionation with two-dimensional gel electrophoresis, we found that a set of host proteins was present in phagosomes that were absent from endosomal/lysosomal compartments. The residence of mycobacteria in compartments outside the endosomal/lysosomal system may explain their persistence inside host cells and their sequestration from immune recognition. Furthermore, the approach described here may contribute to an improved understanding of the molecular mechanisms that determine the intracellular fate of mycobacteria during infection. 相似文献
87.
88.
Therapy of bovine ocular squamous-cell carcinoma with local doses of interleukin-2: 67% complete regressions after 20 months of follow-up 总被引:1,自引:1,他引:0
F. W. Willem Den Otter Graham Hill Wim R. Klein Jan Willem Koten Peter A. Steerenberg Pieter H. M. De Mulder Christopher Rhode Rachel Stewart Joop A. J. Faber E. Joost Ruitenberg Victor P. M. G. Rutten 《Cancer immunology, immunotherapy : CII》1995,41(1):10-14
We have tested the therapeutic potency of peritumorally injected low doses of interleukin-2(IL-2). Seventy tumours of the bovine ocular squamous-cell carcinoma (BOSCC), 1–3 cm in diameter, were treated with 5000, 20 000 or 200 000 U IL-2 from Eurocetus (Chiron) to find the optimal dose for treatment. Injections were given peritumorally on Monday to Friday on 2 consecutive weeks. The size of the tumours was measured before treatment and 1, 3, 4, 9 and 20 months after treatment. After 9 months complete regression was observed in 89% of the tumours treated with 5000 U IL-2, 80% treated with 20 000 U and 67% treated with 200 000 U. After 20 months, there was complete regression of 35%, 31% and 67% of the tumours respectively. The 9-and 20-month results of the 200 000-U treatment are significantly better than those of the 5000-U and 20 000-U treatments taken together. This protocol may be useful to treat advanced inoperable tumours (e.g. of the nasopharynx or skin) of human patients. 相似文献
89.
J A Mol R Docter G Hennemann T J Visser 《Biochemical and biophysical research communications》1984,120(1):28-36
Iodothyronine 5'-deiodinase activity of rat liver microsomes was rapidly and completely lost by treatment with diethylpyrocarbonate (DEP) and by photo-oxidation with Rose Bengal (RB). In both cases inactivation followed pseudo first order reaction kinetics. Inactivation by DEP was diminished in the presence of substrate or competitive inhibitors, and was reversed by hydroxylamine treatment. In addition to photo-oxidation, deiodinase activity was also inhibited by RB in the dark. This inhibition was reversible and competitive with substrate (Ki 60 nM). These results suggest the location of an essential histidine residue at or near the active site of rat liver iodothyronine deiodinase. 相似文献
90.
Summary The effects of disturbing (cultivating) and stockpiling prairie grassland topsoil on microbial activity, microbial biomass C, plant production and decomposition potentials were studied by measuring CO2 efflux from unamended and glucose amended soil in the laboratory and by conducting a pot and litter bag study in the greenhouse. Stockpiling appeared to have very little effect on soil respiratory activity, but did reduce the microbial biomass C levels. Throughout the 3 year study the microbial biomass C in the surface soil of the stockpile was less than that in the undisturbed soil, while the biomass C in soil at the bottom of the stockpile was at no time significantly different from that in the undisturbed soil. The reduction in microbial biomass C in the surface soil immediately after stockpiling was attributed to a decrease in the soil organic C levels caused by a slight dilution of the topsoil with subsurface mineral soil, and the exposure of the stockpile surface to extreme environmental conditions. Soils from all depths of the stockpile responded more slowly to the addition of glucose than soil from the undisturbed and cultivated treatments even when no differences in biomass were detected between the undisturbed and stockpiled soils. It is postulated that the rapidity with which the soil microbial biomass responds to glucose additions may be a sensitive indicator of stress on the soil biological components. The reduction in biomass after storage for 1 year had no adverse effects on the decomposition or primary production potential of the stored soil. Rather, shoot production by fall rye was stimulated in the stored topsoil, presumably a result of better N nutrition. 相似文献