PubMed Health⌕ Search
Home/Archive/PubMed

PubMed · 2428486

Sensitive method for detecting viable cells seeded into bone marrow.

C P Reynolds·A T Black·J N Woody
View original source ↗Export metadata ↓

Abstract

A method for marking viable cells which allows highly sensitive detection of the cells in mixed populations is described. Viable tumor cell lines or normal lymphocytes were stained with the supravital DNA stain Hoechst 33342 (H342) and seeded into human bone marrow. Examination of the marrow mixtures with an inverted fluorescence microscope allows detection of a single H342-stained cell in 1 million marrow cells. H342 staining does not cause significant changes in cell viability or in the sensitivity of the cells to complement mediated lysis. Counterstaining the mixture with trypan blue quenches H342 fluorescence in nonviable cells, limiting detection to only viable cells.

Explore related subjects

BenzimidazolesBone Marrow CellsBone Marrow TransplantationCell SurvivalComplement System ProteinsCytotoxicity, ImmunologicHumansStaining and LabelingJournal ArticleResearch Support, U.S. Gov't, Non-P.H.S.

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

C P Reynolds, A T Black, J N Woody. 1986. Sensitive method for detecting viable cells seeded into bone marrow.. https://pubmed.ncbi.nlm.nih.gov/2428486/

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed1992

Biosynthesis of vitamin B12. Transformation of riboflavin 2H-labeled in the 1'R position of 1'S position into 5,6-dimethylbenzimidazole.

The 5,6-dimethylbenzimidazole moiety of vitamin B12 is formed from riboflavin in aerobic and some aerotolerant bacteria. Thereby C1' of riboflavin is transformed into C2 of the vitamin B12 base. In the present publication a study on this transformation with riboflavin 2H-labeled in the 1'R or 1'S position is described. This study was undertaken in order to find out if one of the two hydrogens at C1' is transferred to C2 of 5,6-dimethylbenzimidazole. The 2H-labeled riboflavin samples were synthesized starting from unlabeled or 1-2H-labeled ribose and 3,4-dimethylaniline yielding N-beta-D-ribopyranosyl-3,4-dimethylaniline. The unlabeled riboside was reduced to N-D-ribityl-3,4-dimethylaniline with sodium cyanoborotrideuteride, the 2H-labeled riboside with sodium cyanoborohydride. The ribityl derivatives were transformed into N-D-ribityl-2-phenylazo-4,5-dimethylaniline, and condensed with barbituric acid to riboflavin. The reduction of the ribosyl compound to the ribityl derivative is only partially stereospecific. Thus the riboflavin synthesized from unlabeled ribose had a 2H ratio of 3/1 (1'R/1'S), the riboflavin obtained from D-[1-2H1]ribose of 1/3 (1'R/1'S). The 2H content in these positions was determined from the 1H-NMR spectra. These spectra showed also that 1 mol 2H/mol riboflavin was present in position 1'. The deuterated riboflavin samples were incubated under aerobic conditions with broken cell preparations of Propionibacterium shermanii. The deuterium content of the 5,6-dimethylbenzimidazole isolated was determined by mass spectrometry and by 1H NMR. These measurements revealed that the hydrogen in the pro-S position at C1' of riboflavin is retained during 5,6-dimethylbenzimidazole formation, and is thus found at C2 of this base.

Benzimidazoles↗
PubMed1992

Identification of 5,6-dimethylbenzimidazole as the Co alpha ligand of the cobamide synthesized by Salmonella typhimurium. Nutritional characterization of mutants defective in biosynthesis of the imidazole ring.

The Co beta-cyano derivative of the cobamide isolated from Salmonella typhimurium was identified as Co alpha-(alpha-5,6-dimethylbenzimidazolyl)-Co beta-cyanocobamide, indicating that this bacterium synthesizes 5,6-dimethylbenzimidazole (DMB) de novo. We found that mutants deficient in the synthesis of DMB can incorporate benzimidazole without modification to form Co alpha-(alpha-benzimidazolyl)cobamide, a cobamide that is physiologically active. The analysis of the nutritional requirements of mutants deficient in DMB synthesis identified 4,5-dimethylphenylenediamine as a putative intermediate in the synthesis of the imidazole ring of DMB. Our results suggest that the CobII region of the cob operon of S. typhimurium only encodes functions involved in the synthesis of the imidazole ring of DMB.

Benzimidazoles↗
PubMed1992

Pyrimido[1,6-a]benzimidazoles: a new class of DNA gyrase inhibitors.

Substituted 4-quinolone- (1, A = CH) and 1,8-naphthyrid-4-one- (1, A = N) 3-carboxylic acids are currently the only classes of clinically useful antibacterial agents exerting their activity by inhibiting the subunit A of DNA gyrase. Pyrimido[1,6-alpha]benzimidazoles 11 have been found to be a new class of inhibitors of this enzyme. The design, synthesis, and biological activity of these compounds are reported.

Benzimidazoles↗