Dissecting cAMP binding domain A in the RIalpha subunit of cAMP-dependent protein kinase. Distinct subsites for recognition of cAMP and the catalytic subunit

The two gene-duplicated cAMP binding domains in the regulatory subunits of cAMP dependent protein kinase are each comprised of an A helix, an eight-stranded beta-barrel, and a B and C helix (1). The A domain is required for high affinity binding to C, while the B domain regulates access to the A domain. Using a combination of a yeast two-hybrid screen coupled with deletion analysis, cAMP binding domain A of RI was dissected into two structurally and functionally distinct subsites, one that binds cAMP and another that binds the C subunit. The minimum stable subdomain required for binding to C in the 1-3 micromolar range is composed of residues 94-169, while residues 236-244, mapped to the C helix of cAMP binding domain A, were defined as a second surface necessary for high affinity (5-10 nanomolar) binding to C. This portion of the C helix, due to its position directly between the two subsites, serves as a molecular switch for either a cAMP-bound conformation or a C-bound conformation and can thus modulate interactions of cAMP binding domain A with cAMP, with C, and with cAMP binding domain B.     

Transluminal balloon angioplasty, stents, and atherectomy

Arterial bypass grafting for occlusive disease is still considered the gold standard in the treatment of arterial occlusive disease. However, less invasive methods are available for select patients. Percutaneous balloon angioplasty has been shown to be effective in focal iliac artery stenosis, with patency rates of 50% to 90% at 5 years. Patency rates for femoropopliteal lesions are generally less than 50% at 2 years. Complications seen with balloon angioplasty often can be treated with arterial stents. Stents can be categorized into balloon-expandable, which provide a rigid scaffold to support the artery, self-expanding, which exert radial force to resist external compression, and thermal expanding stents, which allow for the use of a smaller introducer sheath. Another treatment option is atherectomy, in which the offending lesion is removed instead of fracturing and dilating the lumen. Although overall initial results of atherectomy have not been favorable, short lesions with eccentric atheroma or intimal hyperplasia, such as those seen in dialysis access fistulas or vein graft stenoses, may respond well.     

Aldosterone and dexamethasone stimulate calcineurin activity through a transcription-independent mechanism involving steroid receptor-associated heat shock proteins

1. PD 81,723 has been shown to enhance binding of adenosine to A1 receptors by stabilizing G protein-receptor coupling ('allosteric enhancement'). Evidence has been provided that in the perfused hearts and isolated atria PD 81,723 causes a sensitization to adenosine via this mechanism. 2. We have studied the effect of PD 81,723 in guinea-pig isolated atrial myocytes by use of whole-cell measurement of the muscarinic K+ current (I[K(ACh)]) activated by different Gi-coupled receptors (A1, M2, sphingolipid). PD 81,273 caused inhibition of I[K(ACh)] (IC50 approximately 5 microM) activated by either of the three receptors. Receptor-independent I[K(ACh)] in cells loaded with GTP-gamma-S and background I[K(ACh)], which contributes to the resting conductance of atrial myocytes, were equally sensitive to PD 81,723. At no combination of concentrations of adenosine and PD 81,723 could an enhancing effect be detected. 3. The compound was active from the outside only. Loading of the cells with PD 81,723 (50 microM) via the patch pipette did not affect either I[K(ACh)] or its sensitivity to adenosine. We suggest that PD 81,723 acts as an inhibitor of inward rectifying K+ channels; this is supported by the finding that ventricular I(K1), which shares a large degree of homology with the proteins (GIRK1/GIRK4) forming I[K(ACh)] but is not G protein-gated, was also blocked by this compound. 4. It is concluded that the functional effects of PD 81,723 described in the literature are not mediated by the A1 adenosine receptor-Gi-I[K(ACh)] pathway.     

Transducin-alpha C-terminal peptide binding site consists of C-D and E-F loops of rhodopsin

The binding of heterotrimeric GTP-binding proteins (G-proteins) to serpentine receptors involves several independent contacts. We have deduced the points of interaction between mutant bovine rhodopsins and alphat-(340-350), a peptide corresponding to the C terminus of the alpha subunit (alphat) of bovine retinal G-protein, transducin. Direct binding of alphat-(340-350) to rhodopsin stabilizes the activated metarhodopsin II state (M II), consequently uncoupling the rhodopsin-transducin interaction. This peptide action requires two segments on the cytoplasmic domain of rhodopsin: the Tyr136-Val137-Val138-Val139 sequence on the C-D loop and the Glu247-Lys248-Glu249-Val250-Thr251 sequence on the E-F loop. We propose that a tertiary interaction of these two loop regions forms a pocket for binding the alphat C terminus of the transducin during light transduction in vivo. In most G-proteins, the C termini of alpha subunits are important for interaction with receptors, and, in several serpentine receptors, regions similar to those in rhodopsin are essential for G-protein activation, indicating that the interaction described here may be a generally applicable mode of G-protein binding in signal transduction.     

Evaluation of the effectiveness of continuous renal replacement therapy in critical patients after cardiovascular surgery

The efficacy of continuous methods of renal substitute therapy (RST) in patients with multiple organ failure is assessed. The patients were divided in 2 groups administered different types of PST. Group 1 were 16 patients subjected to RST by peritoneal dialysis, in group 2 (n = 16) GP and/or GDP were used. Hemodynamics, hematological and biochemical values, and clearance of inflammation mediators were monitored and hemohydrobalance and complications of therapy assessed in the course of RST. Both RST methods proved to be highly effective. The possibility of differentiated use of peritoneal dialysis and GP/GDP permits an individual approach to treatment, and equally high efficacy of both methods solves the problem of treating total renal insufficiency in the majority of patients with multiple organ failure following cardiovascular surgery.     

A Practical Cross-Layer Mechanism For Fairness in 802.11 Networks

Many companies, organizations and communities are providing wireless hotspots that provide networking access using 802.11b wireless networks. Since wireless networks are more sensitive to variations in bandwidth and environmental interference than wired networks, most networks support a number of transmission rates that have different error and bandwidth properties. Access points can communicate with multiple clients running at different rates, but this leads to unfair bandwidth allocation. If an access point communicates with a mix of clients using both 1 Mb/s and 11 Mb/s transmission rates, the faster clients are effectively throttled to 1 Mb/s as well. This happens because the 802.11 MAC protocol approximate “station fairness”, with each station given an equal chance to access the media. We provide a solution to provide “rate proportional fairness”, where the 11 Mb/s stations receive more bandwidth than the 1 Mb/s stations. Unlike previous solutions to this problem, our mechanism is easy to implement, works with common operating systems and requires no change to the MAC protocol or the stations. Joseph Dunn received an M.S. in computer science from the University of Colorado at Boulder in 2003, and B. S. in coputer science and mathematics from the University of Arizona in 2001. His research interests are in the general area of computer systems, primarily focusing on security and scalability in distributed systems. He is currently working on his Ph.D. in computer science from the University of Colorado at Boulder. Michael Neufeld received a Ph.D. in Computer Science from the University of Colorado at Boulder in December of 2004, having previously received an M.S. in Computer Science from the University of Colorado at Boulder in 2000 and an A.B. in Computer Science from Princeton University in 1993. His research interests are in the general area of computer system, specifically concentrating on wireless networking, software defind/cognitive radio, and streerable antennas. He is currently a postdoc in the Computer Science department at the University of Calorado at Boulder pursuing research related to software defined radio and new MAC protocols for steerable phase array antennas. Anmol Sheth is a Ph.D. student in Computer Science at the University of Colorado at Boulder. He received his B.S. in Computer Science from the University of Pune, India in 2001. He has been co-leading the development of the MANTIS operating system. He has co-authored three papers include MAC layer protocol design, energy-efficient wireless communication, and adapting communications to mobility. Dirk Grunwald received his Ph.D. from the University of Illinois in 1989 and joined the University of Colorado the same year. His work addresses research and teaching in the broad area of “computer systems”, which includes computer architecture, operating systems, networks, and storage systems. His interests also include issues in pervasive computing, novel computing models, and enjoying the mountains. He is currently an Associate Professor in the Department of Computer Science and in Electrical and Computer Engineering and is also the Director of the Colorado Center for Information Storage. John Bennett is a Professor of Computer Science with a joint appointment in Electrical and Computer Engineering at the University of Colorado at Boulder. He also serves as Associate Dean for Education in the College of Engineering and Applied Science. He joined the CU-Boulder faculty in 2000, after serving on the faculty of Rice University for 11 years. While at Rice, Bennett pioneered a course in engineering design for both engineering and non-engineering students that has been emulated at several universities and high schools. In addition to other teaching awards, Bennett received the Keck Foundation National Award for Engineering Teaching Excellence for his work on this course. Bennett received his Ph.D. in 1988 from the University of Washington. Prior to completing his doctoral studies, he was a U.S. Naval Officer for several years and founded and served as President of Pacific Mountain Research, Inc., where he supervised the design and development of a number of commercial computing systems. Bennett's primary research interests are broadly focused in the area of distributed systems, and more narrowly in distributed information management and distributed robotic macrosensors.     

Traumatic aniridia after small incision cataract extraction.

INTRODUCTION: Phacoemulsification and falls are both common in the elderly population. We present a case of acquired total aniridia and vitreous haemorrhage occurring as a result of trauma in a pseudophakic eye. METHODS: Interventional case report with history, clinical photograph and discussion with literature review. RESULTS: A previously healthy 74-year-old Caucasian female was referred with a painful left eye and poor vision following a fall and trauma to the left side of her face. Initial examination confirmed visual acuity of perception of light with total hyphaema and vitreous haemorrhage. Subsequent examination revealed complete aniridia with an intact capsular bag and well-centred posterior chamber intraocular lens and attached retina. Final visual acuity after treatment was 6/9. CONCLUSIONS: Blunt trauma may cause total iris disinsertion in and expulsion from pseudophakic eyes. This relatively novel injury may present increasingly commonly to casualty departments and ophthalmologists.