This report concerns a carrier cell culture (designated JCI) infected persistently with JC virus (JCV). Immunostaining with an anti-JCV antiserum revealed that JCI was a carrier culture in which only a small fraction of the cells (approximately 1.5%) produced the virus. The JCV titre was increased strikingly by incubating confluent JCI cells for 4-6 days in medium containing a low concentration of fetal bovine serum (2%). Viral genomes cloned from the persistently infected JCI cells were heterogeneous with respect to size, but most clones had an alteration of the same regulatory region (designated CR-JCI). Transfection experiments with a chimeric JCV DNA (Mad-1/CR-JCI), in which the regulatory region was CR-JCI and the other region was derived from an infectious JCV (Mad-1) DNA, showed that CR-JCI was less efficient in inducing viral growth than the regulatory regions of IMR-32-adapted JCVs. The transfected cells could be readily subcultured, and they continued to produce JCV. It is concluded that a decrease in the activity of the JCV regulatory region is of importance for the maintenance of the carrier state of JCI cells.

List of Abbreviations:AIDS, acquired immunodeficiency syndrome; BKV, BK virus; BKVAN, BK virus-associated nephropathy; CPE, cytopathic effect; DAB, 3,3 -diaminobenzidine tetrahydrochloride; FBS, fetal bovine serum; HA, hemagglutination; HIV-1, human immunodeficiency virus-1; JCV, JC virus; LTR, long terminal repeat; luc, luciferase; MEM, minimum essential medium; PBS, phosphate-buffered saline; PCR, polymerase chain reaction; pLTR-luc, HIV-1 LTR-dependent reporter gene; PML, progressive multifocal leukoencephalopathy; PML-type JCV, JCV isolated from the brain of PML patients; pRSV-β gal, RSV promoter-dependent expression vector for β-galactosidase; RLU, relative light units; RT, reverse transcriptase; SV40, simian virus 40.replication origin and agnoprotein gene in JCV genomic DNA. It is well known that the regulatory region of JCV isolated from PML-type JCV contains hypervariable sequences (2).

No abstract

No abstract

Percutaneously absorbed carbon dioxide enhances blood flow. The mechanism by which it does so is unclear, but we hypothesized that it involves bicarbonate ions. BALB/c mice were bathed in neutral bicarbonate ionized water (NBIW) and showed increased blood bicarbonate levels and blood flow via phosphorylation of peripheral vascular endothelial nitric oxide synthase (eNOS) and production of nitric oxide (NO). Phosphorylation of eNOS and NO production were also increased in human umbilical vein endothelial cells cultured in medium containing NBIW, and NBIW showed reactive oxygen species scavenging activity. In a double-blind, randomized study in men and women aged 30 to 59 years with subjective cold intolerance, bathing in NBIW elevated body temperature faster than bathing in a control solution and improved chills and sleep quality. Taken together, our results show that percutaneously absorbed carbon dioxide changes to bicarbonate ions, which act directly on endothelial cells to increase NO production by phosphorylation of eNOS and thus improve blood flow.