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1.
Exp Hematol ; 41(10): 894-902, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23727584

ABSTRACT

Polymerase chain reaction (PCR)-negative molecular complete remission (mCR) can be induced by stem cell transplantation in some patients with multiple myeloma (MM) and is associated with long-term progression-free survival (PFS). The detection of molecular minimal residual disease (MRD), however, requires fresh or frozen materials for designing clone-specific primers, which are not always readily available. In this study, we used DNA extracted from archival bone marrow (BM) slides for PCR to detect MRD in 50 patients with MM who received various induction therapies and autologous peripheral blood stem cell transplantation (ASCT). Clonotype-specific immunoglobulin (Ig) H PCR primers were prepared for 32 of 50 cases (64%) using BM slides, and for 9 of 14 cases (64%) using fresh BM cells. DNA in peripheral blood stem cell autografts of the 22 patients who achieved at least a partial response after ASCT was subjected to PCR to amplify clonotype-specific rearranged IgH gene sequences. The median PFS of the eight patients with MRD-positive autografts was 18 months, whereas that of 14 patients with MRD-negative autografts was not reached at a median follow-up of 27 months (p = 0.012). Post-ASCT PFS of the four patients who achieved mCR was 100% at a median follow-up of 47 months. These results indicate that archival BM slides can serve as a source of DNA for preparing clonotype-specific primers for MRD monitoring in patients with MM whose cryopreserved myeloma cells are not available for DNA preparation. Our results also suggest that patients with MM who received MRD-negative autografts and achieved mCR have a long PFS.


Subject(s)
Bone Marrow/pathology , DNA Primers/genetics , Multiple Myeloma/diagnosis , Neoplasm, Residual/diagnosis , Polymerase Chain Reaction , Adult , Aged , Female , Genes, Immunoglobulin Heavy Chain/genetics , Histocytological Preparation Techniques , Humans , Immunoglobulins/genetics , Male , Middle Aged , Multiple Myeloma/genetics , Neoplasm, Residual/genetics , Predictive Value of Tests , Prognosis , Retrospective Studies
2.
Microbiology (Reading) ; 153(Pt 1): 92-101, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17185538

ABSTRACT

The sigma gene, sigI, of Bacillus subtilis belongs to the group IV heat-shock response genes and has many orthologues in the bacterial phylum Firmicutes. The B. subtilis sigI gene is considered to constitute an operon with rsgI (regulation of sigI, formerly ykrI). As little is known about either the structure and function of the sigI-rsgI operon or the SigI regulons, the role of RsgI in heat-inducible transcription of the sigI-rsgI operon was investigated, using Northern analysis and a heat-stable beta-galactosidase reporter assay. Heat-inducible, SigI-dependent transcription of the sigI-rsgI operon was stimulated greatly by disrupting rsgI. Yeast two-hybrid analysis showed direct interaction between the N-terminal portion of the presumed RsgI protein and SigI. Without RsgI function, induction of transcription of the sigI-rsgI operon upon transient heat stress depended on dnaK activity. However, transcription of the operon was induced during growth at prolonged higher temperature even without DnaK function. Without RsgI function, sigI-rsgI operon transcription was induced after the end of growth independent of any temperature shift in a sporulation medium and toward the end of growth in a rich complex medium. Furthermore, glucose addition resulted in a strong suppression of sigI-rsgI transcription. Therefore it is hypothesized that transcription of the sigI-rsgI operon of B. subtilis is negatively regulated by the putative transmembrane protein RsgI, which moderates SigI's sensitivity to heat shock or nutritional stress.


Subject(s)
Bacillus subtilis/genetics , Bacterial Proteins/genetics , Down-Regulation , Heat-Shock Proteins/genetics , Sigma Factor/genetics , Bacterial Proteins/biosynthesis , Base Sequence , Heat-Shock Response , Molecular Sequence Data , Operon/genetics , Promoter Regions, Genetic/genetics , Transcription, Genetic
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