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991.
L Xu D Sgroi CJ Sterner RL Beauchamp DM Pinney S Keel K Ueki JL Rutter AJ Buckler DN Louis 《Canadian Metallurgical Quarterly》1994,54(20):5262-5264
The CDKN2 gene that encodes the cell cycle regulatory protein cyclin-dependent kinase-4 inhibitor (p16) has recently been mapped to chromosome 9p21. Frequent homozygous deletions of this gene have been documented in cell lines derived from different types of tumors, including breast tumors, suggesting that CDKN2 is a tumor suppressor gene involved in a wide variety of human cancers. To determine the frequency of CDKN2 mutations in breast carcinomas, we screened 37 primary tumors and 5 established breast tumor cell lines by single-strand conformation polymorphism analysis. In addition, Southern blot analysis was performed on a set of five primary breast carcinoma samples and five breast tumor cell lines. Two of the five tumor cell lines revealed a homozygous deletion of the CDKN2 gene, but no mutations were observed in any of the primary breast carcinomas. These results suggest that the mutation of the CDKN2 gene may not be a critical genetic change in the formation of primary breast carcinoma. 相似文献
992.
JL Freeman ML Kreck DJ Uhlinger JD Lambeth 《Canadian Metallurgical Quarterly》1994,33(45):13431-13435
Rac, a small molecular weight GTPase in the Ras superfamily, participates in the activation of the multicomponent superoxide-generating NADPH oxidase of human neutrophils. Rac is 30% identical to Ras overall, but is 75% identical within the sequence corresponding to the effector region of Ras, which regulates mitogenesis through interactions with the protein kinase Raf1. We investigated the role of this region in Rac1 using site-directed mutagenesis. In a cell-free semirecombinant NADPH oxidase system, mutants in the 26, 33, 38, and 45 amino acids showed 20-110-fold reduced binding to the oxidase complex as judged by EC50 values and reduced (44-80%) maximal activities in superoxide generation. Only the GTP gamma S-bound form associated, since the GDP-bound form of Rac neither activated alone nor competed with GTP gamma S-Rac. EC50 values for neither p47-phox nor p67-phox were affected when mutant Racs were used in place of Rac. Data indicate direct binding of the Rac effector region to one or more components of the respiratory burst oxidase. Results indicate a general role for conserved effector-equivalent regions in small GTPases in the regulation of protein-protein interactions. 相似文献
993.
In the absence of L-arginine, the heme center of the oxygenase domain of neuronal nitric-oxide synthase reduces molecular oxygen to superoxide (O-2). Our recent work has provided evidence that inducible NOS (iNOS) may also catalyze O-2 formation in macrophages. However, there has been a lack of direct evidence of superoxide generation from the purified iNOS, and it was previously hypothesized that significant O-2 production does not occur. Moreover, the mechanism and enzyme site responsible for O-2 generation is unknown. To determine whether iNOS produces O-2 and to identify the mechanism of this process, we performed electron paramagnetic resonance measurements on purified iNOS using the spin trap 5,5-dimethyl-1-pyrroline N-oxide. In the presence of NADPH, prominent O-2 adduct signals were detected from iNOS. These signals were totally abolished by superoxide dismutase but not affected by catalase. High concentrations of L-arginine decreased this O-2 formation, whereas its enantiomer D-arginine did not. Pre-incubation of iNOS with the flavoprotein inhibitor diphenyleneiodonium totally blocked these O-2 signals. Conversely, pretreatment of the enzyme with the heme blocker cyanide had no effect on O-2 generation. Furthermore, strong O-2 generation was directly detected from the isolated iNOS reductase domain. Together, these data demonstrate that iNOS does generate O-2, and this mainly occurs at the flavin-binding sites of the reductase domain. 相似文献
994.
995.
ZL Sun DN Fang XY Wu HH Ritchie C Bègue-Kirn JC Wataha CT Hanks WT Butler 《Canadian Metallurgical Quarterly》1998,37(3-4):251-261
The purpose of this study was to characterize the molecular expression of a spontaneously immortalized and cloned cell line (MDPC-23) derived from 18-19 day CD-I fetal mouse molar dental papillae to determine if these cells were odontoblast-like. Western blots showed that a protein band, at approximately 105 kDa, reacting positively with anti-DSP antibodies and co-migrating with mouse DSP, was present in lysates of cells from passages 7, 37 and 77, in serum-free conditioned medium from passage 37 cells, and in mouse dentin extract. A minor band at 55 kDa was also apparent in cell lysates. Using a cDNA probe for a 486bp mouse DSP coding sequence, DSP or DSP-PP mRNA expression was detected by Northern analysis as well as Southern analysis after RT-PCR in all three passages. It was also shown that in these cells 1,25 (OH)2 vitamin D3 upregulated both osteopontin and osteocalcin mRNA, and dexamethasone downregulated alkaline phosphatase and alpha2(I) collagen mRNA. Thus, MDPC-23 cells express proteins which are common to mineralizing tissue. The expression of DSP and DSP-PP strongly suggests that this cell line is from the odontoblast lineage. 相似文献
996.
JE Cobb SG Blanchard EG Boswell KK Brown PS Charifson JP Cooper JL Collins M Dezube BR Henke EA Hull-Ryde DH Lake JM Lenhard W Oliver J Oplinger M Pentti DJ Parks KD Plunket WQ Tong 《Canadian Metallurgical Quarterly》1998,41(25):5055-5069
3-?4-[2-(Benzoxazol-2-ylmethylamino)ethoxy]phenyl?-(2S)-((2- benzoylph enyl)amino)propionic acid (1) and (2S)-((2-benzoylphenyl)amino)-3-?4-[2-(5-methyl-2-phenyloxazol-4-y l)e thoxy]phenyl?propionic acid (2) are peroxisome proliferator-activated receptor gamma (PPARgamma) agonists and have antidiabetic activity in rodent models of type 2 diabetes. As part of an effort to develop the SAR of the N-2-benzoylphenyl moiety of 1 and 2, a series of novel carboxylic acid analogues, 23-66, modified only in the N-2-benzoylphenyl moiety were synthesized from L-tyrosine and evaluated as PPARgamma agonists. In general, only modest changes in the N-2-benzoylphenyl moiety of 1 and 2 are tolerated. More specifically, the best changes involve bioisosteric replacement of one of the two phenyl rings of this moiety. Addition of substituents to this moiety generally produced compounds that are less active in the cell-based functional assays of PPARgamma activity although binding affinity to PPARgamma may be maintained. A particularly promising set of analogues is the anthranilic acid esters 63-66 in which the phenyl ring in the 2-benzoyl group of 1 and 2 has been replaced by an alkoxy group. In particular, (S)-2-(1-carboxy-2-?4-[2-(5-methyl-2-phenyloxazol-4-yl)ethoxy]phen yl? ethylamino)benzoic acid methyl ester (63) has a pKi of 8.43 in the binding assay using human PPARgamma ligand binding domain and a pEC50 of 9.21 in the in vitro murine lipogenesis functional assay of PPARgamma activity. Finally, 63 was found to normalize glycemia when dosed at 3 mg/kg bid po in the Zucker diabetic fatty rat model of type 2 diabetes. 相似文献
997.
Consistency of optimum chromosome spreading during harvest of cytogenetic specimens remains a major concern. We have tested the idea that a precise control of the drying rate (the time with which metaphase cells dry), as fixed cell suspension is placed on a slide or an in situ culture in last fixation, may be the answer. Amniocyte and lymphocyte cultures were allowed to dry at defined combinations of relative humidity (RH) and temperature (T) in a modified Thermotron environmental control unit. We were able to demonstrate, based on 2,250 amniocytes and 1,650 lymphocytes, that the metaphase area after drying was a function of RH and T for both in situ and non-in situ culture systems. As the RH and T increase, the metaphase area increases until a threshold is reached. Also, as RH increases, the slide drying time increases. Data obtained using a response surface regression, proportional hazards regression analysis and slide drying time studies are consistent with our model of chromosome spreading. Optimum metaphase areas can be achieved at various combinations of RH and T. We propose that the use of an environmental control unit is a practical way of achieving optimum chromosome spreading routinely and in a highly consistent manner. 相似文献
998.
999.
RH Wilson JP Preece DL Salamon JL Sperry SP Bornstein 《Canadian Metallurgical Quarterly》1994,5(4):269-277
The Tonal and Speech Materials for Auditory Perceptual Assessment, Disc 1.0 audio compact disc developed in 1992 includes several sets of degraded speech materials, two of which, time compression and reverberation, are described in this paper. The digital techniques used to compress the NU No. 6 materials (female speaker) on an 80386-based computer are described, along with a series of experiments on subjects with normal hearing that document the effects of the time compression on recognition performance. Experiment I examined at 70 dB SPL the effect on word recognition of 45, 55, 65, 70, and 75 percent compressions. Experiment II developed psychometric functions for the 45, 65, and 75 percent time-compression conditions. Experiment III defined the effects that time-compression degradation (45% and 65%) plus reverberation degradation (0.3 sec) had on the recognition performance on the NU No. 6 materials. Based on the experiments, four conditions (45% compression, 65% compression, 45% compression plus 0.3-sec reverberation, and 65% compression plus 0.3-sec reverberation) were selected and recorded on the compact disc. In the compact disc trials, normative data on the four conditions were developed from 120 listeners with normal hearing. 相似文献
1000.
C Leray LL Sarliève H Dreyfus R Massarelli L Binaglia L Freysz 《Canadian Metallurgical Quarterly》1994,29(1):77-81
We examined 4 cases of synovial sarcoma, 3 biphasic-one of which was located in the abdominal wall-and 1 monophasic, in patients between 16 and 54 years. Immunohistochemistry revealed the capacity of these neoplasms to express mesenchymal and epithelial markers, not only in the sarcomatous-like stroma but also in gland-like component. We also found that in the epitheliomorphous cells vimentin positivity had a characteristic basal position whereas EMA and cytocheratins were much more positive in the apical zone. This double positivity could be profitably utilized when the predominance of the gland-like component in a synovial sarcoma requires differential diagnosis from metastatic adenocarcinoma. 相似文献