Rabbit Anti-Human PTK2 Polyclonal Antibody

We prepare an extract of dog urine (DLU) that, when applied to monolayers of MDCK cells (epithelial, derived from a normal dog), enhances the transepithelial electrical resistance (TER) in a dose-dependent manner. This increase is not reflected in variations of the linear amount of TJ nor in changes of the pattern of junctional strands as observed in freeze fracture replicas, nor in the distribution of claudin I (a membrane protein of the TJ) nor ZO-1 (a TJ-associated protein). A preliminary characterization of the active component of DLU indicates that it weighs 30-50 kDa, bears a net negative electric charge, and is destroyed by type I protease but not by 10-min boiling. DLUs prepared from human, dog, rabbit and cat are effective on MDCK cells. However, dog DLU increases TER in MDCK (dog) as well as LLCPK1 (pig) monolayers, but not in other epithelial cell lines such as LLCRK1 (rabbit), PTK2 (kangaroo) and MA-104 (monkey), nor in the endothelial cell line CPA(47) (cow). Given that in its transit from the glomerulus to the urinary bladder the filtrate increases its concentration by more than two orders of magnitude, the substance(s) we report may act at increasingly higher concentrations in each segment, and afford a potential clue to the progressive increase of TER across the walls of the nephron from the proximal to the collecting duct.

Exposure to environmental and lifestyle factors, such as cigarette smoking, affect the epigenome and might mediate risk for diseases and cancers. We have performed a genome-wide DNA methylation study to determine the effect of smoke and snuff (smokeless tobacco) on DNA methylation. A total of 95 sites were differentially methylated [false discovery rate (FDR) q-values 0.05] in smokers and a subset of the differentially methylated loci were also differentially expressed in smokers. We found no sites, neither any biological functions nor molecular processes enriched for smoke-less tobacco-related differential DNA methylation. This suggests that methylation changes are not caused by the basic components of the tobacco but from its burnt products. Instead, we see a clear enrichment (FDR q-value 0.05) for genes, including CPOX, CDKN1A and PTK2, involved in response to arsenic-containing substance, which agrees with smoke containing small amounts of arsenic. A large number of biological functions and molecular processes with links to disease conditions are also enriched (FDR q-value 0.05) for smoke-related DNA methylation changes. These include insulin receptor binding, and negative regulation of glucose import which are associated with diabetes, positive regulation of interleukin-6-mediated signaling pathway, regulation of T-helper 2 cell differentiation, positive regulation of interleukin-13 production which are associated with the immune system and sertoli cell fate commitment which is important for male fertility. Since type 2 diabetes, repressed immune system and infertility have previously been associated with smoking, our results suggest that this might be mediated by DNA methylation changes.