Improvement permeabilization

Labalette-Houache, M., Torpier, G., Capron, A., and Dessaint, J. P. (1991) Improved permeabilization procedure for flow cytometric detection of internal antigens. Analysis of interleukin-2 production. J. Immunol. Methods 138,143-153. 

Schmid, 1., Uittengobaart, C. H., and Giogi, J. V. (1991) A gentle fixation and permeabilization method for combined cell-snrface and intracellular staining with improved precision in DNA qnantification. Cytometry 12, 279-285. 

The limitations of the application of conventional detergents mentioned above can be circumvented by replacing this approach with cell membrane permeabilization by microwave heating. Improved detection of intracellular antigens can be obtained with microwave heating used in combination with flow cytometry. This approach yields histogram patterns that show clear discrimination between intact cells and cell debris (Fig. 9.5). 

In parallel with the fermentative process, Genencor has developed an in vitro process to convert Glc into 2,5DKG [22, 168]. This latter system encompasses a redox couple of free GlcDH and 2,5DKGR that mediate the first and last steps in the route, complemented by GADH and 2KGDH in a permeabilized P. citrea (Fig. 8.31). Its virtues are said to be a quantitative yield of 2KLG without byproducts, improved STY and simple DSP [22]. 

Picher etal, 1994). This ATPase could be substantially ( 70% in rabbit portal vein) reduced by pretreatment of the preparation by the ectonucleotidase inhibitor 4,4 -diisothiocyanatostilbene-2,2 -disulfonic acid (DIDS), followed by inclusion of sodium azide in solutions used for permeabilized preparations. This treatment dramatically reduced the ADP/ATP ratio measured in permeabilized muscle and increased contractile force. This method may greatly improve the usefulness of a-toxin-permeabilized preparations for energetics studies. 

Insulin is generally administered parenterally (subcutaneous injection), although many researchers and companies are developing alternative administration methods, i.e., oral, buccal, (Oral-lyn) or pulmonary (Exubera) [10], These methods require greatly increased dosage in the case of buccal or pulmonary administration or additives to improve the bioavailability of orally administered insulin (including encapsulation, permeabilization, or chemical stabilization additives). No intesti-nally absorbed (oral) version of insulin has yet been proven effective and received FDA approval. 

Huang, Y, Rubinsky, B., 1999. Micro-electroporation improving the efficiency and understanding of electrical permeabilization of cells. Biomed. Microdevices 2, 145—150. 

On the other hand, treatment of E. coli with the peptide antibioticpolymyxin B led to an effective permeabilization of E. coli supporting the entry of abietic acid, which led to an almost fivefold improved conversion of abietic acid into 15-hydroxyabietic acid by CYP105A1 [92]. 

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