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V M Subbotin

Publications and source records attributed to V M Subbotin.

71 records · Page 4Linked to original sources
PubMedDate not supplied

Time course of gene expression after plasmid DNA gene transfer to the liver.

BACKGROUND: High levels of expression in hepatocytes can be achieved after intraportal delivery of plasmid DNA vectors with up to 10% of all liver cells transfected. CMV promoter-driven expression is very high on Day 1 after injection, but is diminished strongly by Day 2. Expression slowly declines after 1 week. We describe experiments aimed at elucidating the reasons for this rapid decline in transgene expression. METHODS: Histological methods were used to determine the presence and extent of liver damage and hepatocyte proliferation. Viral and liver-specific promoters were tested to study promoter shut-off, Southern blotting was performed to determine the loss of the pDNA vector over time, and several mouse models were used to study the host immunological response. RESULTS: pDNA is lost rapidly early after injection, but remains at a relatively stable copy number after Day 4. Southern blotting experiments showed that plasmid DNA could be detected for at least 12 weeks after injection (0.2 copies per genome). The early rapid decline of expression is promoter dependent. A liver-specific albumin promoter resulted in similar levels of expression on Days 1 and 7, suggesting that promoter inactivation may be responsible for the instability of CMV promoter-driven expression. The slow decline in expression levels after 1 week appears to be the result of an immune response directed against the expressed transgene. Expression was much prolonged in immunosuppressed, immunodeficient, or antigen-tolerized mice. CONCLUSION: The present data suggest that if promoter inactivation can be overcome, intravascular delivery of plasmid DNA could be a highly efficient, simple and non-toxic liver gene therapy approach. Intravascular delivery of pDNA allows for the rapid screening of novel expression vectors in vivo.

Animals↗
PubMedDate not supplied

[Changes in the functional state and structure of the adrenals in CC14 poisoning].

Introduction of CCI4 in doses of 0.1 and 0.2 ml/100 g to albino rats reduced the steroid production generated by adrenal sections. In 24 and 48 hours after poisoning with a small dose of CCI4 (0.1 ml/100 g) there could be seen an increased concentration of 11-oxycorticosteroids in the plasma. Morphological signs of a damage suffered by the gland were observed only in poisoning with a large dose of CCI4 (0.2 ml/100 g).

11-Hydroxycorticosteroids↗