Cellular mutants define a common mRNA degradation pathway targeting cytokine AU-rich elements.

To functionally classify AU-rich elements (AREs) from six different cytokine mRNAs, we made use of two previously described HT1080-derived cellular mutants (slowA, slowC) that lack a function required for the rapid degradation of interleukin-3 (IL-3) mRNA. Here we show that the defect is specific for ARE-containing mRNAs, whereas nonsense-mediated decay is intact. Degradation of beta-globin reporter transcripts mediated by the AREs of IL-3, GM-CSF, and TNFalpha, as well as by the structurally different and less potent AREs of IL-2 and IL-6, is impaired in both mutants. All these reporter transcripts are also sensitive to decay induced by ectopic expression of the RNA-binding protein tristetraprolin in the slowC background. Thus, we concluded that the mutants slowA and slowC define a common mRNA degradation pathway that targets cytokine AREs. In NIH3T3 cells, this decay pathway becomes incapacitated by upstream signaling from p38 MAP- or PI3-kinases, which independently stabilize cytokine ARE-containing transcripts. In contrast, c-fos ARE-directed mRNA degradation proceeds through a different pathway not affected by these kinases

Angiotensin II is an endogenous neurotransmitter for rat and human mesenteric resistance blood vessels

Angiotensin II (Ang II) is one of the most potent vasoconstrictors. We document here the innervation of rat and human mesenteric resistance arteries (MRA) by angiotensinergic neurons of the rat and human sympathetic coeliac ganglia. Angiotensinogen (Ang-N)-mRNA and angiotensin converting enzyme-mRNA but no renin-mRNA were detected by using quantitative real time polymerase chain reaction in total RNA extracts of rat coeliac ganglia. In the same extracts, cathepsin D-mRNA was detected: This protease also cleaves Ang I from Ang-N and could therefore account for the generation of neuronal Ang peptides in the absence of renin. In situ hybridization confirmed the presence of Ang-N-mRNA in the cytoplasm of rat coeliac ganglia. By using solid-phase extraction, high performance liquid chromatography and subsequent radioimmunoassay, Ang II and its metabolites were detected in rat and also in human coeliac ganglia. Immunoreactivity for Ang II was demonstrated in rat and human coeliac ganglia neurons and their projections innervating MRA. In addition, segmental angiotensinergic innervation of MRA was also observed. By means of confocal laser scanning microscopy we were able to demonstrate the presence of angiotensinergic synapses en passant along side of vascular smooth muscle cells. Our findings could indicate that Ang II is synthesized inside the neurons of sympathetic coeliac ganglia and may act as an endogenous neurotransmitter locally in MRA

Swiss Community Pharmacies' on the Web and Pharmacists' Experiences with E-commerce: Longitudinal study and Internet-based questionnaire survey

BACKGROUND: There are multiple ways in which community pharmacies can present themselves on the Internet, e.g., as a platform for drug information or as an advertising platform for their services. OBJECTIVE: To estimate the number of Swiss community pharmacies on the Internet over the period of 32 months (2000-2003), to describe their current e-commerce services, and to explore the experiences and plans these pharmacies have with regard to their Internet presence. METHODS: A longitudinal study was performed to determine the number of Swiss German pharmacies on the Internet by conducting Internet searches in 2000, 2001, and 2003. In April 2002, a cross-sectional Internet-based survey was administered to explore the pharmacies' experiences and plans regarding their Web sites. RESULTS: As of April 2003, 373 (44%) of 852 community pharmacies from the German speaking part of Switzerland were on the Internet. One hundred eighty four listed an e-mail address and were asked to complete a questionnaire. Of the 107 pharmacies answering the survey questions (58% response rate): 46% had been on the Internet for 1 to 2 years; 33% of the Web sites are part of a pharmacy group's Web portal; 31% of the pharmacies plan to expand their Internet appearance in the future; 74% provide e-commerce services, with 81% of those pharmacies filling five or less orders per month; and 12% plan on expanding their e-commerce services in the future. CONCLUSIONS: The number of community pharmacies offering Internet services steadily increased over 32 months. Given the importance of the Internet as a tool for information, communication, and advertising for pharmacy products and services, it can be expected that the increase will continue. Pharmacy-group portals are important promoters of pharmacies on the Internet. For many community pharmacies, Internet portals that provide an Internet presence for the pharmacies and provide regularly-updated content (e.g., health news, tips, drug information) seem to be the most effective solutions. Even though 40% of the pharmacies already offer e-commerce services, these services are still of minor importance. For many pharmacists, the current legal regulations seem to be unclear. Most pharmacies want to maintain their Internet services

Molecular analysis of SMARD1 patient-derived cells demonstrates that nonsense-mediated mRNA decay is impaired

no abstract availabl