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21.
CM Lucas ED Franke MI Cachay A Tejada ME Cruz RD Kreutzer DC Barker SH McCann DM Watts 《Canadian Metallurgical Quarterly》1998,59(2):312-317
Asymmetric acetylcholinesterase (AChE) is anchored to the basal lamina (BL) of cholinergic synapses via its collagenic tail, yet the complement of matrix receptors involved in its attachment remains unknown. The development of a novel overlay technique has allowed us to identify two Torpedo BL components that bind asymmetric AChE: a polypeptide of approximately 140 kDa and a doublet of 195-215 kDa. These were found to stain metachromatically with Coomassie blue R-250, were solubilized by acetic acid, and were sensitive to collagenase treatment. Upon sequence analysis, the 140 kDa polypeptide yielded a characteristic collagenous motif. Another AChE-binding BL constituent, identified by overlay, corresponded to a heparan sulfate proteoglycan. Lastly, we established that this proteoglycan, but not the collagenous proteins, interacted with at least one heparin binding domain of the collagenic tail of AChE. Our results indicate that at least two BL receptors are likely to exist for asymmetric AChE in Torpedo electric organ. 相似文献
22.
JM Sorof EP Hawkins ED Brewer II Boydstun AS Kale DR Powell 《Canadian Metallurgical Quarterly》1998,12(9):764-768
In patients with proteinuria, African-American (AA) ethnicity is reported to be a risk factor for focal segmental glomerulosclereosis (FSGS) and its progression to end-stage renal disease (ESRD). We reviewed our single-center experience to determine the probability of FSGS and its progression to ESRD based on ethnicity and age at presentation in children with proteinuria with or without nephrotic syndrome. Proteinuria without systemic disease or acute glomerulonephritis was the presenting feature in 17% (236/1,403) of children in the renal patient database of Texas Children's Hospital, Baylor College of Medicine. Histopathological diagnoses were established in 107 of 236 patients (45%). FSGS was identified in 65 patients, accounting for 28% of all patients with proteinuria and 61% of patients who underwent renal biopsy. FSGS was more prevalent in AA (45%) than in non-AA patients (22%) (P=0.001), and AA patients with FSGS were older at presentation (12.7+/-4.4 years) than non-AA patients (5.6+/-4.6 years) (P<0.001). Among patients who underwent renal biopsy, increasing age at presentation increased the probability of having FSGS in AA but not non-AA patients (P=0.04). Five-year actuarial renal survival of FSGS was worse in AA (8%) than in non-AA patients (31%) (P=0.01). These data suggest an increased risk and worse outcome of FSGS in AA compared with non-AA children. 相似文献
23.
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An intrinsic feature of yeast artificial chromosomes (YACs) is that the cloned DNA is generally in the same size range (i.e., approximately 200-2000 kb) as the endogenous yeast chromosomes. As a result, the isolation of YAC DNA, which typically involves separation by pulsed-field gel electrophoresis, is frequently confounded by the presence of a comigrating or closely migrating endogenous yeast chromosome(s). We have developed a strategy that reliably allows the isolation of any YAC free of endogenous yeast chromosomes. Using recombination-mediated chromosome fragmentation, a set of Saccharomyces cerevisiae host strains was systematically constructed. Each strain contains defined alterations in its electrophoretic karyotype, which provide a large-size interval devoid of endogenous chromosomes (i.e., a karyotypic "window"). All of the constructed strains contain the kar1-delta 15 mutation, thereby allowing the efficient transfer of a YAC from its original host into an appropriately selected window strain using the kar1-transfer procedure. This approach provides a robust and efficient means to obtain relatively pure YAC DNA regardless of YAC size. 相似文献
25.
ED Cox H Diaz-Arauzo Q Huang MS Reddy C Ma B Harris R McKernan P Skolnick JM Cook 《Canadian Metallurgical Quarterly》1998,41(14):2537-2552
A pharmacophore and an alignment rule have previously been reported for BzR agonist ligands. The design and synthesis of 6-(propyloxy)-4-(methoxymethyl)-beta-carboline-3-carboxylic acid ethyl ester (6-PBC, 24, IC50 = 8.1 nM) was based on this pharmacophore. When evaluated in vivo this ligand exhibited anticonvulsant/anxiolytic activity but was devoid of the muscle relaxant/ataxic effects of "classical" 1,4-benzodiazepines (i.e., diazepam). Significantly, 6-PBC 24 also reversed diazepam-induced muscle relaxation in mice. The 3-substituted analogues 40-46 and 48 of 6-PBC 24 and Zk 93423 27(IC50 = 1 nM) were synthesized and evaluated in vitro to determine what affect these modifications would have on the binding affinity at recombinant BzR subtypes. With the exception of the 3-amino ligands 40 and 41, all the beta-carbolines were found to exhibit high binding affinity at BzR sites. The 3-propyl ether derivative 45 was also evaluated in vivo and found to be devoid of any proconvulsant or anticonvulsant activity at doses up to 40 mg/kg. The 6-(1-naphthylmethyloxy) and 6-octyloxy analogues 25, 26, 28, and 29 of 6-PBC 24 were synthesized to further evaluate the proposed alignment of agonists vs inverse agonists in the pharmacophore of the BzR. In addition, ligands 26 and 29 were designed to probe the dimensions of lipophilic pocket L3 at the agonist site. The activity of 29 was evaluated in vivo; however, this analogue elicited no pharmacological effects at doses up to 80 mg/kg. These and other related beta-carbolines were also examined in five recombinant GABAA receptor subtypes. Ligands 52-61 all exhibited moderate to high affinity at GABAA receptors containing alpha1 subunits. These ligands will be useful in further defining the pharmacophore at alpha1 beta3 gamma2 receptors. 相似文献
26.
ED Simmons TG Burke T Haley J Medige 《Canadian Metallurgical Quarterly》1996,21(3):295-8; discussion 299
STUDY DESIGN: This study evaluates and compares the stiffness of two cervical spine fixation techniques. OBJECTIVES: This biomechanical study was carried out to compare the interspinous and Dewar cervical spine fixation techniques. SUMMARY OF BACKGROUND DATA: Interspinous wiring is a commonly used method of fixation in the cervical spine. The Dewar technique is less commonly known and practiced, and clinical experience has suggested that it may be a more stable technique. METHODS: Cervical spine specimens stabilized with the interspinous and "Dewar" techniques were biomechanically tested in flexion and in torsion. Stiffness and energy absorption under moderate loads were compared. The Dewar technique uses contoured double corticocancellous iliac grafts as internal grafts/splints fixed to the spine with threaded pins and wire. The interspinous technique is a single interspinous wire loop. Eleven fresh human cervical spines were harvested from cadavers. The spines were destabilized at C4-C5 by sectioning all tissue except the anterior longitudinal ligament. Each fixation technique was applied alternatively and tested on each spine. RESULTS: In torsion testing (n = 5), the Dewar fusion was 61% stiffer than the interspinous technique (P < 0.02). Dewar: 11.3 N/mm (s.d. 4.9 N/mm) and interspinous: 8.4 N/mm (SD 3.3 N/mm). In flexion testing (n = 6), the Dewar technique was 35% stiffer than the interspinous technique (P < 0.10). Dewar: 655.4 Nmm/degree (SD 293 Nmm/degree) and interspinous: 406.8 Nmm/degree (SD 113.0 Nmm/degree). Energy absorption with the interspinous technique was greater in flexion (P < 0.10) and in torsion (P < 0.005), indicating more deformation with the interspinous technique. There was no statistically significant difference between the means of specimens tested first and those tested second independently of the fixation technique. CONCLUSIONS: These tests indicate that the Dewar cervical spine fixation is stiffer than the single interspinous wire in both flexion and particularly torsion. This project is the only biomechanical study of the Dewar technique that we are aware of, and the results support the clinical findings regarding the effectiveness of this technique. 相似文献
27.
BK Yoder WG Richards C Sommardahl WE Sweeney EJ Michaud JE Wilkinson ED Avner RP Woychik 《Canadian Metallurgical Quarterly》1996,50(4):1240-1248
Autosomal recessive polycystic kidney disease (ARPKD) is characterized by the formation of large collecting tubule and ductular cysts that often result in renal insufficiency within the first decade of life. Understanding the process leading to cyst formation will require the identification and characterization of genes involved in the etiology of this disease. In this regard, we previously described the generation of a mouse model (TgN737Rpw) for ARPKD and the cloning of a candidate gene. Here we show direct involvement of the Tg737 gene in collecting duct cyst formation by expressing the wild-type Tg737 cDNA as a transgene in TgN737Rpw mutants. In contrast to TgN737Rpw mutants, the "rescued" animals survive longer, have normal renal function and normal localization of the EGFr to the basolateral surfaces of collecting duct epithelium. 相似文献
28.
Inhibition of HeLa cell messenger RNA translation by 7-methylguanosine 5'-monophosphate 总被引:2,自引:0,他引:2
LA Weber ER Feman ED Hickey MC Williams C Baglioni 《Canadian Metallurgical Quarterly》1976,251(18):5657-5662
Translation of HeLa cell RNA containing poly(A) in a wheat germ cell-free system is markedly but incompletely inhibited by 7-methylguanosine 5'-monophosphate (m7G5'p). We have analyzed the translation products synthesized in the presence of different concentrations of m7G5'p and find that translation of all mRNAs is equally inhibited. To demonstrate the specificity of the inhibitor for RNAs with 5'-terminal m7G5' ppp... we show that specific translation products of satellite tobacco necrosis virus RNA, which does not have this 5' terminus, are synthesized in the presence of m7G5' p. Protein synthesis programmed by endogenous mRNA in a HeLa cell-free system is inhibited after a 10-min lag by m7G5' p. Other guanosine nucleotides without the 7-methyl group or with the phosphate in a different position are not inhibitor. We show that translation of all mRNAs is inhibited to a similar extent by m7G5'p in the HeLa cell-free system, by synthesizing 35S-labeled proteins in the presence of different inhibitory concentrations of this nucleotide and analyzing the translation products by electrophoresis and autoradiography. Translation of encephalomyocarditis virus RNA added to the HeLa cell-free system is not inhibited by m7"g5p; this viral RNA does not have this nucleotide at the 5' terminus. This indicates that m7G5'p specifically inhibits translation of mRNAs with the 5' terminus m7G5'ppp... and suggests that initiation of translation of picornavirus RNA may proceed via a mechanism different from that of cellular mRNAs. 相似文献
29.
30.
NA Lisitsyn MV Rosenberg GA Launer LL Wagner VK Potapov TB Kolesnik ED Sverdlov 《Canadian Metallurgical Quarterly》1993,(3):26-29
We describe a novel technique for isolation of sequences that are present in one genome (tracer), but absent in another (driver). Tracer DNA, cleaved with Sau 3A and capped with a single stranded PCR adapter, is allowed to hybridize with an excess of sheared biotinylated driver; biotinylated DNA and its hybrids with the tracer are removed by phenol/chloroform extraction after incubation with streptavidin. After several rounds of subtraction the ends of self-annealed tracer molecules from the nonextractable fraction are filled-in with Tag polymerase and amplified, using the single stranded PCR adapter as a primer. The method has been applied to purification of fragments from a 2.9 kb plasmid added to E. coli DNA at equimolar quantity. Plasmid derived fragments (250-1000 bp), initially comprising 1/1400th part of tracer DNA, were purified to homogeneity after two rounds of subtraction followed by PCR. 相似文献