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991.
992.
Philip D. Fernsten Jan K. Czyzyk Toshihide Mimura John B. Winfield 《Molecular biology reports》1994,20(2):85-95
Patients with SLE develop IgM autoantibodies to different isoforms of CD45, the major surface membrane protein tyrosine phosphatase on lymphocytes and other nucleated hemopoietic cells. Because such autoantibodies could have a potential role in the development of immune dysfunction in this disorder, we performed a series of experiments to characterize their antigenic specificity further. Blots of recombinantE. coli fusion proteins encoded by exons 3–7 of the p220 and p180 isoforms were uniformly non-reactive with SLE IgM, suggesting that anti-CD45 autoantibodies in SLE are directed against conformational and/or carbohydrate epitopes, rather than linear polypeptide epitopes. This issue was examined further using chemically and enzymatically modified CD45 purified from T cells by lectin affinity chromatography as substrates. Treatment of CD45 with 25 mM sodium-m-periodate, sufficient to abrogate binding to various lectins, abolished the reactivity with SLE anti-CD45 autoantibodies. On the other hand, digestion of CD45 with neuraminidase enhanced the binding of anti-CD45 autoantibodies from some of the SLE sera. This result probably reflects decreased steric hindrance or charge repulsion because the binding of mouse monoclonal antibodies directed against linear polypeptide epitopes of CD45 was similarly enhanced. Digestion of CD45 with N-glycosidase F had no effect on autoantibody staining. Taken together, these data suggest that IgM anti-CD45 autoantibodies in SLE recognize non-sialylated carbohydrate determinants in the highly O-glycosylated polymorphic domains of CD45.Abbreviations SLE
systemic lupus erythematosus
- SBA
soybean agglutinin
- RCAI
Ricinus communis agglutinin
- SNL
Sambucus nigra lectin
- MBP
maltose binding protein
- mAb
monoclonal antibody
- WGA
wheat germ agglutinin 相似文献
993.
Sajal Chakraborti Sandip K. Batabyal John R. Michael Tapati Sanyal 《Molecular and cellular biochemistry》1994,130(2):121-127
Exposure of rabbit pulmonary arterial smooth muscle cells to 10 M of the calcium ionophore A23187 dramatically stimulates cell membrane-associated phospholipase A2 activity and arachidonic acid release. In addition, A23187 also enhances cell membrane-associated serine esterase activity. Serine esterase inhibitors phenylmethylsulfonylfuoride and diisopropyl fluorophosphate prevent the increase in serine esterase and phospholipase A2 activities and arachidonic acid release caused by A23187. A23187 still stimulated serine esterase and phospholipase A2 activities and arachidonic acid release in cells pretreated with nominal Ca2+ free buffer. Treatment of the cell membrane with A23187 does not cause any appreciable change in serine esterase and phospholipase A2 activities. Pretreatment of the cells with actinomycin D or cycloheximide did not prevent the increase in the cell membrane associated serine esterase and phospholipase A2 activities, and arachidonic acid release caused by A23187. These results suggest that (i) a membrane-associated serine esterase plays an important role in stimulating the smooth muscle cell membrane associated phospholipase A2 activity (ii) in addition to the presence of extracellular Ca2+, release of Ca2+ from intracellular storage site(s) by A23187 also appears to play a role in stimulating the cell membrane-associated serine esterase and phospholipase A2 activities, and (iii) the increase in the cell membrane-associated serine esterase and phospholipase A2 activities does not appear to require new RNA or protein synthesis.Abbreviations A23187
calcium ionophore
- AA
arachidonic acid
- PMSF
phenylmethyl sulfonylfuoride
- DFP
diisopropyl-fluorophosphate
- DMEM
Dulbecco's modified Eagles medium
- FCS
fetal calf serum
- PBS
phosphate buffered saline
- HBPS
Hank's buffered physiological saline
- PLA2
phospholipase A2 相似文献
994.
Iturin A: A potential new fungicide for stored grains 总被引:6,自引:0,他引:6
The removal of many synthetic fungicides from the market has created a demand for new, environmentally safe fungicides. Iturin A, a cyclic lipopeptide produced byBacillus subtilis, has strong antifungal properties and low mammalian toxicity. To determine the efficacy of this compound as a potential fungicide on stored feed grains, lots of corn, peanuts and cottonseed were treated with varying concentrations of iturin A. The mycoflora of treated seed was assayed along with that of untreated seed and seed treated with fungicides used commercially for planting seed. Fungal species varied considerably in their sensitivity to iturin A. Significant reductions in total mycoflora occurred in most seed lots tested at iturin A concentrations of 50 to 100 ppm. 相似文献
995.
Use of Genetic and Physical Mapping to Locate the Spinal Muscular Atrophy Locus between Two New Highly Polymorphic DNA Markers 总被引:9,自引:2,他引:7
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Olivier Clermont Philippe Burlet Lydie Burglen Suzie Lefebvre Fabrice Pascal John McPherson John J. Wasmuth Daniel Cohen Denis Le Paslier Jean Weissenbach Mark Lathrop Arnold Munnich Judith Melki 《American journal of human genetics》1994,54(4):687-694
The gene for autosomal recessive forms of spinal muscular atrophy (SMA) has recently been mapped to chromosome 5ql3, within a 4-cM region between the blocks D5S465/D5S125 and MAP-1B/D5S112. We identified two new highly polymorphic microsatellite DNA markers—namely, AFM265wf5 (D5S629) and AFM281yh9 (D5S637)—which are the closest markers to the SMA locus. Multilocus analysis by the location-score method was used to establish the best estimate of the SMA gene location. Our data suggest that the most likely location for SMA is between locus D5S629 and the block D5S637/D5S351/MAP-1B/D5S112/D5S357. Genetic analysis of inbred SMA families, based on homozygosity by descent and physical mapping using mega-YACs, gave additional information for the loci order as follows: cen–D5S6–D5S125/D5S465–D5S435–D5S629–SMA–D5S637–D5S351–MAP–1B/D5S112–D5S357–D5S39–tel. These data give the direction for bidirectional walking in order to clone this interval and isolate the SMA gene. 相似文献
996.
997.
Natural and Electroporation-Mediated Transformation of Methanococcus voltae Protoplasts 总被引:3,自引:1,他引:2
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Girishchandra B. Patel John H. E. Nash Brian J. Agnew G. Dennis Sprott 《Applied microbiology》1994,60(3):903-907
The lack of high-efficiency transformation systems has severely impeded genetic research on methanogenic members of the kingdom Archaeobacteria. By using protoplasts of Methanococcus voltae and an integration vector, Mip1, previously shown to impart puromycin resistance, we obtained natural transformation frequencies that were about 80-fold higher (705 transformants per μg of transforming DNA) than that reported with whole cells. Electroporation-mediated transformation of M. voltae protoplasts with covalently closed circular Mip1 DNA was possible, but at lower frequencies of ca. 177 transformants per μg of vector DNA. However, a 380-fold improvement (3,417 transformants per μg of DNA) over the frequency of natural transformation with whole cells was achieved by electroporation of protoplasts with linearized DNA. This general approach, of using protoplasts, should allow the transformation of other methanogens, especially those that may be gently converted to protoplasts as a result of their tendency to lyse in hypotonic solutions. 相似文献
998.
999.
The behaviour and distribution of adultCoccinella septempunctata L. (Coleoptera: Coccinellidae) were recorded in two plots of winter wheat infested with the cereal aphidsSitobion avenae (F.) andMetopolophium dirhodum (Walk.) (Homoptera: Aphididae). One plot was sprayed with the pyrethroid insecticide deltamethrin at a rate of 6.25 g a.i./ha
and the other was left unsprayed. Single ladybird beetles were released sequentially on the ground at the centre of the sprayed
and unsprayed plots and their behaviour and position in the crop canopy were recorded at 30 second intervals for a total of
15 min per beetle. Assessments, with fresh beetles, continued for four days after spray application with a total of eighty
ladybird beetles observed. The 15 min period was selected to avoid lethal effects and no ladybird beetles were killed or knocked
down as a result of exposure to deltamethrin residues during this period. Significant differences were found between the overall
behaviour patterns ofC. septempunctata in the untreated and deltamethrin treated plots up to three days after the spray application. Ladybird beetles exposed to
deltamethrin residues were observed to walk and groom significantly more frequently and to rest significantly less frequently
than those in the unsprayed plot. Significant differences were also found between the observed distribution of ladybird beetles
in the sprayed and unsprayed crop canopies, with higher numbers of observations towards the bottom of the crop canopy and
on the ground in the deltamethrin treated plot than in the untreated plot during the first two days after deltamethrin application.
Upon the foliage itself, ladybird beetles were observed significantly more frequently on the abaxial leaf surface in the deltamethrin
treated crop compared with the untreated crop. The results are discussed in terms of possible evidence for the repellency
of deltamethrin toC. septempunctata and also the implications for integrated pest management of changes in predator behaviour and crop distribution resulting
from sub-lethal uptake of insecticides. 相似文献
1000.
Darrell L. Ellsworth Rodney L. Honeycutt Nova J. Silvy John W. Bickham W. D. Klimstra 《Evolution; international journal of organic evolution》1994,48(1):122-136
Mitochondrial DNA (mtDNA) was used to characterize patterns of geographic variation among white-tailed deer (Odocoileus virginianus) populations in the southeastern United States. Fifteen restriction enzymes were employed to survey and map 99 restriction sites in 142 deer from 18 localities in five southeastern states. Phylogenetic analysis revealed three primary groups of haplotypes: (1) southern Florida and the Florida Keys, (2) the remainder of peninsular Florida northward to South Carolina, and (3) the Florida panhandle westward to Mississippi. Geographical heterogeneity in haplotype frequencies suggests that stochastic lineage sorting or isolation by distance are not important determinates of mtDNA differentiation among deer populations. The pattern of mtDNA variation in white-tailed deer is concordant spatially with those observed in unrelated taxa suggesting the common influence of historical biogeographic events. The data (1) support previous hypotheses that relate contemporary patterns of intraspecific phylogeography in northern Florida to the physiogeographic history of the region; and (2) suggest that genetic differentiation in southern Florida may be attributable to episodes of Pleistocene dispersal. Despite potentially high vagility and human intervention, ecological and demographic characteristics of deer have effectively preserved the historical pattern of intraspecific mtDNA differentiation. 相似文献