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81.
Santos Carlos Chandler Karen Zimmer Stephen Fisher Paul B. Gunthert Ursula Anderson Kimberly Ward 《Cell biochemistry and biophysics》1995,26(1):1-19
A parallel-plate flow chamber was used to quantify the detachment of normal cloned rat embryo fibroblasts (CREF) fibroblasts,ras-transformed CREF fibroblasts (CREF T24), and CREF T24 fibroblasts transfected with a Krev/RAP1A suppressor gene (HK B1) from
a confluent monolayer of normal CREF fibroblasts to determine if the expression patterns of CD44 variants (mol wt 110 and
140 kDa) corresponded with detachment properties and metastatic potential. In the detachment assay, known shear stresses ranging
from 20–24 dyn/cm2 were applied to the adherent cells and the number of cells detached from the monolayer after 180 s was determined. Results
showed that cellular expression of CD44 variants correlated with the metastatic potential of the cells and with the cells’
ability to detach from a monolayer of normal cells. Western blot analysis showed a low level of expression of the CD44 variants
in the normal cell line, CREF, and the lowly metastatic cell line, HK B1. Detachment studies showed a low percentage of detachment
of both of these cell lines from a normal cell monolayer. Tumor-derived (HK B1-T) and lung nodule-derived (HK B1-M) cell lines
were established and both formed tumors and metastasis with reduced latency periods as compared to HK B1, but still showed
a markedly delayed latency period compared to the highly metastatic cell line, CREF T24. Both of these cell lines showed a
higher expression of the CD44 variants as compared to CREF and HK B1, and detached easier than CREF and HK B1. CREF T24 showed
a much higher level of expression of the variants and had a higher percentage detachment than all other cell lines. To further
test the role of the CD44 variants in the ability of the cells to detach from the normal monolayer, CREF cells were transfected
with a DNA construct that constitutively expresses the CD44 variants and the detachment properties of three randomly selected
clones were studied. Clones 2 and 3 showed a low level of expression of the CD44 variants after transfection and detached
from the normal monolayer similar to CREF. Clone 1 showed a high level of expression of the CD44 variants and the detachment
of these cells was significantly higher than CREF. From these results, it is concluded that in the five cell lines studied,
expression of the CD44 variants play a significant role in the ability of the cells to detach from a monolayer of normal cells.
It is hypothesized that this detachment may be an important component of a cell’s ability to metastasize. 相似文献
82.
L. M. Freitas dos Santos A. G. Livingston 《Applied microbiology and biotechnology》1994,42(2-3):421-431
An extractive membrane bioreactor has been used to treat a synthetic waste-water containing a toxic volatile organic compound, 1,2-dichloroethane (DCE). Biofilms growing on the surface of the membrane tubes biodegrade DCE while avoiding direct contact between the DCE and the aerating gas. This reduces air stripping by more than an order of magnitude (from 30–35% of the DCE entering the system to less than 1%) relative to conventional aerated bioreactors. Over 99% removal of DCE from a waste-water containing 1600 mg l–1 of DCE was achieved at waste-water residence times of 0.75 h. Biodegradation was verified as the removal mechanism through measurements of CO2 and chloride ion evolution in the bioreactor. No DCE was detected in the biomedium over the operating period. The diffusion-reaction phenomena occurring in the biofilm have been described by a mathematical model, which provides calculated solutions that support the experimental results by predicting that all DCE is biodegraded within the biofilm. Experimentally, however, the rate of DCE degradation in the biofilm was found to be independent of O2 concentration, while the model predictions point to O2 being limiting. 相似文献
83.
MethodsforHandy,RapidIsolationofHighQualityRNAbyGuanadiumThioeyauateDuJianYuanYanhuaMaShenglinDongZhiwei(BeijingInstituteforCancerResearchBeijing100034)硫氰胍是一种有效的蛋白变性剂。早在1979年,Chirgwin等就利用氯化铯/硫氰胍超离心技术成功地从RNA酶富集的胰脏组织中提取出未降解的RNA分子[2],从而使它成为抑制RNA酶的首选药物并得到广泛使用,但受到超速离心设备的限制。1983年,Cathala报道了氯化锂/硫氰胍RNA提取法[1]。该方法操作简便,获得的RNA质量很高,但所需时间较长。为了能在短时间内更快… 相似文献
84.
P. St George-Hyslop E. Rogaeva J. Huterer T. Tsuda J. Santos J. L. Haines K. Schlumpf E. I. Rogaev Y. Liang D. R. Crapper McLachlan J. Kennedy J. Weissenbach G. D. Billingsley D. W. Cox A. E. Lang J. R. Wherrett 《American journal of human genetics》1994,55(1):120-125
A locus for Machado-Joseph disease (MJD) has recently been mapped to a 30-cM region of chromosome 14q in five pedigrees of Japanese descent. MJD is a clinically pleomorphic neurodegenerative disease that was originally described in subjects of Azorean descent. In light of the nonallelic heterogeneity in other inherited spinocere-bellar ataxias, we were interested to determine if the MJD phenotype in Japanese and Azorean pedigrees arose from mutations at the same locus. We provide evidence that MJD in five pedigrees of Azorean descent is also linked to chromosome 14q in an 18-cM region between the markers D14S67 and AACT (multipoint lod score +7.00 near D14S81). We also report molecular evidence for homozy-gosity at the MJD locus in an MJD-affected subject with severe, early-onset symptoms. These observations confirm the initial report of linkage of MJD to chromosome 14; suggest that MJD in Japanese and Azorean subjects may represent allelic or identical mutations at the same locus; and provide one possible explanation (MJD gene dosage) for the observed phenotypic heterogeneity in this disease. 相似文献
85.
dos Santos Silva Edinaldo N. Kakkassery Francy K. Maas Sibylle Dumont Henri J. 《Hydrobiologia》1994,288(2):119-128
Calanoid copepods, belonging to the new genus and species Keraladiaptomus rangareddyi, were collected from temporary ponds in Kerala State, India. The new genus belongs to the family Diaptomidae, subfamily Diaptominae. It is described in detail and its affinity to the related genera, Arctodiaptomus Kiefer, 1932 and Eodiaptomus Kiefer, 1932, discussed. 相似文献
86.
Diel variability in catch rate of juvenile flatfish on two small nursery grounds (Port Erin Bay, Isle of Man and Porto Pirn Bay, Faial, Azores) 总被引:1,自引:0,他引:1
R. D. M. Nash R. S. Santos † A. J. Geffen G. Hughes T. R. Ellis 《Journal of fish biology》1994,44(1):35-45
Variation in beach seine catches was examined in two juvenile flatfish populations (Port Erin, Isle of Man and Porto Pirn, Azores) to determine the importance of diel cycles while controlling for tidal rhythmicity. At Port Erin sampling was undertaken at each low water on consecutive days over a 2-week period in May/June and September 1991. There was no periodicity in the catches of plaice ( Pleuronectes platessa ) in May/June, but in September catches were higher at night. Other flatfish species ( Limanda limanda, Pleuronectes flesus, Scophthalmus maximus and S. rhombus ) sampled at Port Erin were nocturnal. At Porto Pirn, Azores, fish were sampled over 24 h each month from July 1989 to June 1990 and at each low water over a 2-week period in July 1991. The only flatfish species present was Bothuspodas , which had a diurnal pattern in catches over a 1-year period. Sampling at low water in July gave high catches at both midnight and midday. 相似文献
87.
J. B. dos Santos J. Nienhuis P. Skroch J. Tivang M. K. Slocum 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,87(8):909-915
Genetic similarity among 45 Brassica Oleracea genotypes was compared using two molecular markers, random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphisms (RFLPs). The genotypes included 37 broccolis (var. italica), five cauliflowers (var. botrytis) and three cabbages (var. capitata) which represented a wide range of commercially-available germplasm, and included open-pollinated cultivars, commercial hybrids, and inbred parents of hybrid cultivars. Fifty-six polymorphic RFLP bands and 181 polymorphic RAPD bands were generated using 15 random cDNA probes and 62 10-mer primers, respectively. The objectives were to compare RFLP and RAPD markers with regard to their (1) sampling variance, (2) rank correlations of genetic distance among sub-samples, and (3) inheritance. A bootstrap procedure was used to generate 200 random samples of size n (n=2,3,5,... 55) independently from the RAPD and RFLP data sets. The coefficient of variance (CV) was estimated for each sample. Pooled regressions of the coefficient of variance on bootstrap sample size indicated that the rate of decrease in CV with increasing sample size was the same for RFLPs and RAPDs. The rank correlation between the Nei-Li genetic similarity values for all pairs of genotypes (990) based on RFLP and RAPD data was 0.745. Differences were observed between the RFLP and RAPD dendrograms of the 45 genotypes. Overlap in the distributions of rank correlations between independent sub-samples from the RAPD data set, compared to correlations between RFLP and RAPD sub-samples, suggest that observed differences in estimation of genetic similarity between RAPDs and RFLPs is largely due to sampling error rather than due to DNA-based differences in how RAPDs and RFLPs reveal polymorphisms. A crossing algorithm was used to generate hypothetical banding patterns of hybrids based on the genotypes of the parents. The results of this study indicate that RAPDs provide a level of resolution equivalent to RFLPs for detemination of the genetic relationships among genotypes. 相似文献
88.
We have analyzed a total of 12 different global and local multiple
protein-sequence alignment methods. The purpose of this study is to
evaluate each method's ability to correctly identify the ordered series of
motifs found among all members of a given protein family. Four
phylogenetically distributed sets of sequences from the hemoglobin, kinase,
aspartic acid protease, and ribonuclease H protein families were used to
test the methods. The performance of all 12 methods was affected by (1) the
number of sequences in the test sets, (2) the degree of similarity among
the sequences, and (3) the number of indels required to produce a multiple
alignment. Global methods generally performed better than local methods in
the detection of motif patterns.
相似文献
89.
E. A. Santos R. Keller 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1993,163(5):374-379
The effects of glutamate, aspartate, glycine, proline, alanine, taurine, glycerol, glucose and lactate injections on the haemolymph levels of the crustancean hyperglycemic hormone and/or glucose and lactate in the shore crab, Carcinus maenas, were investigated. Only glucose and lactate caused significant changes of hyperglycaemic hormone levels. Glucose injections resulted in a drop of both hormone and lactate, while lactate had an opposite effect, i.e. it raised both crustacean hormone and glucose levels. The results suggest that during increases in glycolytic flux, lactate may cause a release of hormone by a positive feedback mechanism. The hormone would then stimulate glycogenolysis, thus increasing glucose availability. If more glucose is released than is metabolized, excess glucose may leak from the cells and suppress crustancean hyperglycemic hormone release from the X-organ/sinus gland complex by negative feedback.Abbreviations ABTS
2,2-azino-bis (3-ethylbenzthiazoline sulphonic acid)
- ANOVA
one-way analysis of variance
- BSA
bovine serum albumin
- BW
body weight
- CHH
crustacean hyperglycemic hormone
- ELISA
cnzyme-liked immunosorbent assay
- GIH
gonadinhibiting hormone
- IgG
immunoglobin G
- MIH
moult-inhibiting hormone
- MTGXO
medulla terminalis X-organ
- PB
sodium phosphate buffer
- PBS
phosphate buffered saline
- Pi
inorganic phosphate
- XO-SG
X-organ-sinus gland complex 相似文献
90.
L. M. Freitas dos Santos A. G. Livingston 《Applied microbiology and biotechnology》1993,40(1):151-157
The biological treatment of waste-waters containing 1,2-dichloroethane (DCE) in conventional bioreactors results in air-stripping of DCE. In the present work, a novel bioreactor system intended to overcome this problem has been developed for the treatment of a synthetically concocted DCE-containing waste-water (1000 mg DCE l–1). The operation of a conventional air-lift bioreactor at a waste-water flow rate of 0.24 l h–1 led to 33% of the DCE supplied to the reactor being lost to the exit gas stream. The use of the novel enclosed system, operated with a recycling O2 sparge instead of air, resulted in negligible air-stripping at the same waste-water flow rate. A control system was implemented to add O2 as required to maintain the pressure of the recycle gas stream, and a scrubber removed the CO2 produced. Over 99% of DCE supplied was biodegraded during operation of this system, and virtually all carbon entering the system was evolved as CO2.
Correspondence to: A. G. Livingston
Correspondence to: A. G. Livingston 相似文献