Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.64 (proteinase K)
4,071 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Human monocytes kill Actinomycin D-treated WEHI 164 sarcoma cells in a 6 h 51Cr release assay (drug dependent cellular cytotoxicity, DDCC). In the present study we have investigated and characterized the human monocyte production of a cytotoxic factor which mediates DDCC. Cell-free supernatants obtained culturing monocytes for 4-5 h kill Actinomycin D-treated WEHI 164 cells but not untreated tumor cells. A series of antiproteases inhibits the cytotoxic activity of cell-free monocyte supernatants, whereas scavengers of reactive oxygen intermediates were ineffective. The lytic activity was destroyed treating supernatants at 100 degrees C for 5 min or by exposure to acid pH or to proteinase K, whereas it was unaffected by heating at 56 degrees C for 30 min. Upon gel filtration on Sephacryl S200, cytolytic activity eluted in the 33,000 molecular weight range.
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PMID:Rapid killing of actinomycin D-treated tumor cells--cytotoxicity of cell-free monocyte supernatants. 409 54

The polymerase chain reaction was used for Moloney murine sarcoma virus (MoMuSV) detection in frozen and formalin-fixed, paraffin-embedded tissue sections and cultured cells isolated from MoMuSV-induced tumors. Rapid DNA extraction by proteinase K digestion, followed by CHROMA SPIN + TE-100 column purification proved to be satisfactory. Two pairs of overlapping primers, flanking 1026 base pair (bp) to 221 bp, allowed to choose among four different length of DNA-amplified segments. Although net amplification was obtained for frozen tissue and tumor cultured cells in all combinations of primers, the maximum specificity and sensitivity resulted with 602 bp fragment. This product was fully and adequately digestible using Apa I and Sau3A I restriction endonucleases. DNA extracted from paraffin-embedded sections yielded an amplification product only when the primer pair which delineated a 221-bp segment was used. This reproducible method could be useful for diagnostic and for pathogenetic investigations of MoMuSV infections.
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PMID:Detection of Moloney murine sarcoma virus in tissues and cultured cells by the polymerase chain reaction. 809 98