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91.
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JC Neal S Stolnik E Schacht ER Kenawy MC Garnett SS Davis L Illum 《Canadian Metallurgical Quarterly》1998,87(10):1242-1248
Poloxamer 407 and poloxamine 908 have been used by many research groups to modify the surface of both model latex and biodegradable nanospheres, thereby producing nanospheres that have shown reduced protein adsorption in vitro and extended circulation times in vivo. A potential limitation of such systems is the desorption of the copolymer coating layer. We describe a two-stage process to radiolabel poloxamer 407 and poloxamine 908 that has facilitated an investigation into this potential desorption, in vitro. The first stage of the labeling procedure involved the substitution of the terminal hydroxyl groups in each poly(ethylene oxide) (PEO) chain of poloxamer 407 and poloxamine 908 with an amino group. The aminated copolymers were then radiolabeled with 125Iodine Bolton-Hunter reagent. The efficiency of labeling was calculated to be approximately 20% for the tetramine poloxamine 908 and approximately 33% for the diamine poloxamer 407. Remaining free amino groups were then either acetylated, using acetic anhydride, or left in the free amino form. Covalent linkage of the radiolabel to the copolymer was confirmed by nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. The stability of the link between radiolabel and copolymer to hydrolysis was also confirmed; <4% loss of radiolabel occurred from poloxamine 908 after incubation in phosphate-buffered saline (PBS) at 37 degrees C for 8 days. The radiolabeled copolymers (with the free amino groups acetylated) were then used in experiments that have given the first direct evidence that adsorbed copolymers can be displaced by serum proteins in significant amounts from the surface of model and biodegradable nanospheres. The displacement was highly dependent on copolymer-nanosphere compatibility, with up to 78% of 125I tetramine poloxamine 908 being displaced from poly(lactide-co-glycolide) (PLGA) nanospheres in 24 h, compared with 20% displacement of 125I tetramine poloxamine 908 in 24 h from polystyrene nanospheres. These results have direct implication for the future design of drug delivery systems based on coated nanospheres. 相似文献
94.
J Wayne H Suh Z Misulovin KA Sokol K Inaba MC Nussenzweig 《Canadian Metallurgical Quarterly》1994,1(2):95-107
Effects of etoposide (VP-16) and cytosine arabinoside (Ara-C) on the cell cycle of HL-60 and THP-1 cells were studied by flow cytometry using the bromodeoxyuridine (BrdU)/DNA assay technique to investigate the efficacy of VP-16 for monocytic leukemia cells. VP-16 inhibited the proliferation of THP-1 cells more strongly than that of HL-60 cells at any concentrations used at 24 and 48 hr. VP-16 arrested HL-60 and THP-1 cells in the G2/M phase and reduced them in the G0/G1 and early S phase at higher concentrations. There was no significant difference in the percentage of G2/M phase cells at the same concentration between both cells. However, reduction in the G0/G1 and early S phase cells was more marked in THP-1 than HL-60 cells significantly. On the other hand, Ara-C perturbed the cell cycle of HL-60 cells more than that of THP-1 cells at 24 and 48 hr. These results suggest that the effects of VP-16 on the cell cycle may be more intense in THP-1 than HL-60 cells, and support the efficacy of VP-16 for treating monocytic leukemia in vivo. 相似文献
95.
In the dry milling of wheat flour, each unit process (roller mill, purifier, sifter, etc.) produces a mixture with varying amounts of wheat endosperm and non-endosperm byproducts. Chemical images with 82 000 pixels of each intermediate product stream issuing from an individual processing machine are readily analyzed in terms of the relative amount of endosperm and non-endosperm. Approximately three minutes is required to produce an image of each intermediate product stream. Applying partial least squares (PLS) chemometric software to identify individual pixels, which enables calculation of the relative amount of endosperm and non-endosperm, is not a time-limiting factor. When relative flow rates are known for each stream, mass balance can be calculated from each intermediate stream in terms of the product (endosperm content) and the lower value non-endosperm byproduct. Data is presented from a purifier in a commercial flour mill. Intermediate streams collected from a run with optimized operational parameters were compared to those of another run before adjustment. The endosperm (product) mass balance profile for each run enabled assessment of operational efficiency. The devised chemical imaging analysis system would be particularly useful in commissioning of a new mill or to optimize existing wheat milling systems. Also, when raw material differs from that for which previous optimization was established, a new optimization may be in order. The ability to acquire a large number of spectra from a specimen and apply multivariate statistics to identify each pixel and subsequently count pixels accommodates heterogeneity and reports the results from averaging a very large number of individual spectra. A second illustration of the utility of the imaging method is presented centering on streams from the first and second break unit operations at the beginning of the roller mill process. 相似文献
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97.
Networks containing both flexible segments and rigid structures were synthesized on the basis of bisphenol A novolacs and diglycidylether of butanediol using imidazole as an accelerator. A stoichiometric ratio between epoxy groups and phenolic groups of the novolacs leads to networks with methylene bridges as network junctions. In contrast to this, the same reaction with bisphenol A leads to completely soluble products. The glass transition temperature of this soluble material is considerably lower than the glass transition temperature of the networks. Increasing content of methylene bridges in the novolacs leads to an increase of the glass transition temperature of the networks and to a decrease of the δcp value at the glass transition. Furthermore, epoxy excess leads to networks with rubber-structure of the bisphenol A novolac used in the reaction with the diglycidylether. It was found that conformations with intramolecular hydrogen bondings exist between phenolic hydroxyl groups, which considerably influence the reactivity of the novolac with the epoxy group. © 1996 John Wiley & Sons, Inc. 相似文献
98.
The susceptibilities of 143 Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens isolates to 18 antimicrobial agents were tested. All P. gingivalis isolates were susceptible. In contrast, some Prevotella spp. (17%) were resistant to beta-lactams, erythromycin, clindamycin, or tetracycline and carried resistance genes, ermF or tetQ, or beta-lactamases. 相似文献
99.
100.
Antisense oligonucleotide inhibition of hepatitis C virus gene expression in transformed hepatocytes
R Hanecak V Brown-Driver MC Fox RF Azad S Furusako C Nozaki C Ford H Sasmor KP Anderson 《Canadian Metallurgical Quarterly》1996,70(8):5203-5212
Genetic and biochemical studies have provided convincing evidence that the 5' noncoding region (5' NCR) of hepatitis C virus (HCV) is highly conserved among viral isolates worldwide and that translation of HCV is directed by an internal ribosome entry site (IRES) located within the 5' NCR. We have investigated inhibition of HCV gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon, and core protein coding sequences. Oligonucleotides were evaluated for activity after treatment of a human hepatocyte cell line expressing the HCV 5' NCR, core protein coding sequences, and the majority of the envelope gene (E1). More than 50 oligonucleotides were evaluated for inhibition of HCV RNA and protein expression. Two oligonucleotides, ISIS 6095, targeted to a stem-loop structure within the 5' NCR known to be important for IRES function, and ISIS 6547, targeted to sequences spanning the AUG used for initiation of HCV polyprotein translation, were found to be the most effective at inhibiting HCV gene expression. ISIS 6095 and 6547 caused concentration-dependent reductions in HCV RNA and protein levels, with 50% inhibitory concentrations of 0.1 to 0.2 microM. Reduction of RNA levels, and subsequently protein levels, by these phosphorothioate oligonucleotides was consistent with RNase H cleavage of RNA at the site of oligonucleotide hybridization. Chemically modified HCV antisense phosphodiester oligonucleotides were designed and evaluated for inhibition of core protein expression to identify oligonucleotides and HCV target sequences that do not require RNase H activity to inhibit expression. A uniformly modified 2'-methoxyethoxy phosphodiester antisense oligonucleotide complementary to the initiator AUG reduced HCV core protein levels as effectively as phosphorothioate oligonucleotide ISIS 6095 but without reducing HCV RNA levels. Results of our studies show that HCV gene expression is reduced by antisense oligonucleotides and demonstrate that it is feasible to design antisense oligonucleotide inhibitors of translation that do not require RNase H activation. The data demonstrate that chemically modified antisense oligonucleotides can be used as tools to identify important regulatory sequences and/or structures important for efficient translation of HCV. 相似文献