Lithiasis of the distal ureter: ESWL or URS

Human herpesvirus 8 (HHV-8) is a newly discovered virus closely associated with Kaposi's sarcoma and primary effusion lymphomas. When they occur in patients with AIDS, these B-cell lymphomas frequently harbor another human herpesvirus, Epstein-Barr virus (EBV). To determine the molecular mechanisms of the regulation of early gene expression by the immediate-early gene products of HHV-8 and to assess possible molecular interactions between HHV-8 and EBV, we studied the regulation of the HHV-8 thymidine kinase (TK) promoter in cell lines harboring either or both viruses. The constitutive chloramphenicol acetyltransferase (CAT) activity of the TK promoter was low in all six cell lines tested. A putative immediate-early gene product of HHV-8 ORF50, which is a homolog of EBV BRLF1, was cloned into an expression vector and tested for its transactivating capacity. In the presence of 12-O-tetradecanoyl-phorbol-13-acetate (TPA), the CAT activity of the TK promoter was increased 7- to 720-fold by cotransfection with the ORF50 clone in EBV-producing cell lines (Ramos/AW, P3HR-1, and BC-1) but not in EBV-negative cell lines (BCBL-1 and Ramos), nor in the latently EBV-infected cell line Raji. The TK promoter contains three consensus SP1- and two AP1-binding sites. In electrophoretic mobility shift assays, the cellular factor SP1, but not AP1, was found to bind specifically to the TK promoter. To determine whether the increased CAT activity resulted from the interaction of SP1 with the ORF50 gene product, we introduced mutations into two SP1-binding sites. Both mutated SP1 sites had reduced SP1-binding activity and greatly decreased TK promoter responsiveness to ORF50 transactivation, suggesting that upregulation of TK promoter by ORF50 is SP1 dependent.     

Unique regulation of immediate early gene and tyrosine hydroxylase expression in the odor-deprived mouse olfactory bulb

Tyrosine hydroxylase (TH), expressed in a population of periglomerular neurons intrinsic to the olfactory bulb, displays dramatic down-regulation in response to odor deprivation. To begin to elucidate the importance of immediate early genes (IEG) in TH gene regulation, the present study examined expression of IEGs in the olfactory bulb in response to odor deprivation. In addition, the composition of TH AP-1 and CRE binding complexes was investigated in control and odor-deprived mice. Immunocytochemical studies showed that c-Fos, Fos-B, Jun-D, CRE-binding protein (CREB), and phosphorylated CREB (pCREB) are colocalized with TH in the dopaminergic periglomerular neurons. Unilateral naris closure resulted in down-regulation of c-Fos and Fos-B, but not Jun-D, CREB, or pCREB, in the glomerular layer of the ipsilateral olfactory bulb. Gel shift assays demonstrated a significant decrease (32%) in TH AP-1, but not CRE, binding activity in the odor-deprived bulb. Fos-B was found to be the exclusive member of the Fos family present in the TH AP-1 complex. CREB, CRE modulator protein (CREM), Fos-B, and Jun-D, but not c-Fos, all contributed to the CRE DNA-protein complex. These results indicated that Fos-B, acting through both AP-1 and CRE motifs, may be implicated in the regulation of TH expression in the olfactory bulb dopaminergic neurons.