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141.
142.
Gary C. du Moulin Zorina Pitkin Yuan-Jin Shen Evelyn Conti Jean Ko Stewart Carla Charles Dylan Hamilton 《Cytotechnology》1994,15(1-3):365-372
Somatic cell and gene therapy involve the application of biological technologies to an individual patient through the use of living cells which provide a therapeutic benefit (Aliski, 1991). Various forms of cellular and gene therapies are being developed and evaluated in an increasing number of clinical trials for congential and acquired disorders. The potential and progress of these therapeutic applications have resulted in an increasing effort by the Food and Drug Administration (FDA) to develop the regulatory framework under which these therapeutic approaches would insure safety and efficacy, the primary mandate of the FDA.Over five years ago Cellcor began to define the parameters, specifications, and conditions relevant to a Quality Assurance/Quality Control (QA/QC) program that has evolved to insure safety and maximize the efficacy of applications of the company'sex vivo technology, autolymphocyte therapy. Autolymphocyte therapy is an outpatient form of somatic cell immunotherapy based upon the infusion of T cells that have been activatedex vivo using a combination of previously generated autologous cytokines and an anti-CD3 monoclonal antibody.We have been able to demonstrate the feasibility for the safe, controlled, and consistent preparation and delivery of a cellular therapy by application of relevant GMP regulations. This presentation reviews aspects of this program and chronicles our experience which at present amounts to over 4400 infusions for over 700 patients. This program provides a high degree of assurance that a cellular therapy program can be carried out in a multisite mode involving hundreds of patients through the strict adherence to cGMP as set forth in existing regulations. It would be prudent that developers of cellular andex vivo gene therapies establish a similar cell processing and QA/QC infrastructure at an early developmental stage to optimize safety and reproducibility and facilitate regulatory review. 相似文献
143.
Soluble methane monooxygenase (sMMO) maximization studies were carried out as part of a larger effort directed towards the development and optimization of an aqueous phase, multistage, membrane bioreactor system for treatment of polluted groundwater. A modified version of the naphthalene oxidation assay was utilized to determine the effects of methane:oxygen ratio, nutrient supply, and supplementary carbon sources on maximizing and maintaining sMMO activity inMethylosinus trichosporium OB3b.Methylosinus trichosporium OB3b attained peak sMMO activity (275–300 nmol of naphthol formed h–1 mg of protein–1 at 25°C) in early stationary growth phase when grown in nitrate mineral salts (NMS) medium. With the onset of methane limitation however, sMMO activity rapidly declined. It was possible to define a simplified nitrate mineral salts (NMS) medium, containing nitrate, phosphate and a source of iron and magnesium, which allowed reasonably high growth rates (max 0.08 h–1) and growth yields (0.4–0.5 g cells/g CH4) and near maximal activities of sMMO. In long term batch culture incubations sMMO activity reached a stable plateau at approximately 45–50% of the initial peak level and this was maintained over several weeks. The addition of d-biotin, pyridoxine, and vitamin B12 (cyanocobalamin) increased the activity level of sMMO in actively growing methanotrophs by 25–75%. The addition of these growth factors to the simplified NMS medium was found to increase the plateau sMMO level in long term batch cultures up to 70% of the original peak activity.Abbreviations sMMO
soluble methane monooxygenase
- pMMO
particulate methane monooxygenase
- NMS
nitrate mineral salts
- TCE
trichloroethene
- NADH
reduced nicotinamide adenine dinucleotide 相似文献
144.
Life-history traits of the lizard Sceloporus undulatus from two populations raised in a common laboratory environment 总被引:3,自引:0,他引:3
Hatchling Sceloporus undulatus elongatus from Washington Co., Utah and S. u. garmani from Woods Co., Oklahoma were raised to maturity and reproduction under identical laboratory conditions with ad libitum food availability. Growth, allometry, age and size of maturity, clutch size and egg mass were compared among lab-raised cohorts from the two populations, among lab-raised and field-caught animals (including their field-caught mothers) and, for growth, with values obtained by previously published field studies on the same or nearby populations. For all traits population differences observed in previous field studies and current field samples resulted from both a plastic response to proximate environmental conditions and intrinsic (possibly genetic) difference. The most plastic traits were growth and age of maturity. Cohorts from both populations expressed the ability to mature in less than 6 months in the laboratory but only the S.u. garmani express early maturity in the field. Allometric differences generated during growth in the lab were not observed in field samples but may reflect an adaptive physiological difference. The least plastic trait was egg mass. The only trait for which the rank order of the difference in the field was reversed in the lab was growth rate. S.u. elongatus grew significantly faster than S.u. garmani in the lab but much slower in the field. The tendency of S.u. garmani females to breed at minimum size of maturity may be greater than that of S.u. elongatus. 相似文献
145.
Gary R. Jacobson Cynthia Saraceni-Richards 《Journal of bioenergetics and biomembranes》1993,25(6):621-626
The bacterial phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS) consists of several proteins whose primary functions are to transport and phosphorylate their substrates. The complexity of the PTS undoubtedly reflects its additional roles in chemotaxis to PTS substrates and in regulation of other metabolic processes in the cell. The PTS permeases (Enzymes II) are the membrane-associated proteins of the PTS that sequentially recognize, transport, and phosphorylate their specific substrates in separate steps, and theEscherichia coli mannitol permease is one of the best studied of these proteins. It consists of two cytoplasmic domains (EIIA and EIIB) involved in mannitol phosphorylation and an integral membrane domain (EIIC) which is sufficient to bind mannitol, but which transports mannitol at a rate that is dependent on phosphorylation of the EIIA and EIIB domains. Recent results show that several residues in a hydrophilic, 85-residue segment of the EIIC domain are important for the binding, transport, and phosphorylation of mannitol. This segment may be at least partially exposed to the cytoplasm of the cell. A model is proposed in which this region of the EIIC domain is crucial in coupling phosphorylation of the EIIB domain to transport through the EIIC domain of the mannitol permease. 相似文献
146.
Antisera towards neurotensin (NT) and the structurally related peptide, LANT6, were used to characterize immunoreactive peptides and proteins in extracts of chicken tissues. A 17 kDa protein was identified by Western blotting as a potential precursor to NT and LANT6. However, the posttranslational processing of this common precursor appeared to be tissue specific, giving rise to disproportionate amounts of NT and LANT6, along with varying expression of a large molecular LANT6 (Mr, 15 kDa). The intestinal cells containing immunoreactive NT, LANT6, and large molecular LANT6 behaved similarly during fractionation by size and density. These activities also banded together in particles resembling vesicles during centrifugation of isotonic homogenates of tissue. These results suggest that chicken NT and LANT6 are biosynthesized as parts of the same precursor, the processing of which can give rise to a variety of products stored within secretory vesicles. 相似文献
147.
Gary Brown 《Plant Ecology》1994,115(1):77-90
The vegetation at various sites within two separate areas (Mechernich and Aachen) of the Eifel Mountains, Germany/Belgium, both characterized by elevated concentrations of heavy metals in their soils, was surveyed in order to investigate the relationships between soil chemical attributes and floristic composition. In both areas, the typical heavy metal communities can form distinct zones, clearly separated from the surrounding heavy metal-sensitive vegetation. However, an intergrading of heavy metal-tolerant and-sensitive vegetation types is not uncommon and such overlaps can occupy large areas. In Mechernich, soil toxicity is primarily determined by the effects of lead, which is best expressed in terms of the Pb/Ca ratio rather than the absolute levels of this metal in the soil. Soils of heavy metal-sensitive vegetation types have a low Pb/Ca ratio, whereas it is considerably higher in areas supporting heavy metal vegetation. Zinc appears to exert little influence on the floristic composition of the investigated vegetation types. In Aachen, zinc is the predominant heavy metal determining vegetation development. Absolute zinc levels of soils do not accurately reflect zinc toxicity. Analogous to the role of the Pb/Ca ratio in the Mechernich area, the Zn/Ca ratio not only separates heavy metal-sensitive and highly tolerant vegetation units, but also gives a good indication of the gradient operating between the two vegetation types. Lead is probably only of local importance in influencing species composition. 相似文献
148.
Glucocorticoids have been shown to be essential for the excessive fat deposition and development of obesity in several animal models. This study was performed to characterize the role of glucocorticoids in the developmental regulation of adipose tissue metabolism. On day 70 of gestation, pig fetuses were hypophysectomized by micro-cauterization. Hypophysectomized fetuses were implanted subcutaneously with hydrocortisone pellets or received no hormone replacement. Fetuses were removed by laparotomy on day 90 of gestation. Additional fetuses were hypophysectomized on day 70, implanted with hydrocortisone pellets on day 90 and removed on day 105 of gestation. Several intact fetuses were also implanted subcutaneously with hydrocortisone pellets during this later gestational period. Serum cortisol concentrations were reduced in hypophysectomized pigs at both fetal ages and were restored to intact levels by hydrocortisone treatment. Hydrocortisone supplementation enhanced lipolytic response to isoproterenol in intact fetuses but failed to restore lipolytic response to isoproterenol in hypophysectomized animals at either fetal age. Hydrocortisone induced a slight increase in lipogenesis in hypophysectomized fetuses when administered from 70 to 90 days of gestation and a more dramatic increase when administered from days 90 to 105 of gestation. However, hydrocortisone had no effect on basal or insulin stimulated lipogenesis in intact fetuses when administered from days 90 to 105 of gestation. These results indicate that hydrocortisone may have a primary influence on adipose tissue metabolism during late fetal development only in the absence of inhibition from counterregulatory hormones of pituitary origin. 相似文献
149.
An eosin Y staining technique that permits detection of various proteins, including membrane sialoglycoproteins, in polyacrylamide gels is described. The sensitivity of the eosin Y staining method is comparable to silver staining. In addition, there is an added advantage of the antigen icily of the stained proteins being retained in a Western blot. Details of the procedure to obtain optimal staining results are described. 相似文献
150.