Regulation of sorting and post-Golgi trafficking of rhodopsin by its C-terminal sequence QVS(A)PA

Several mutations that cause severe forms of the human disease autosomal dominant retinitis pigmentosa cluster in the C-terminal region of rhodopsin. Recent studies have implicated the C-terminal domain of rhodopsin in its trafficking on specialized post-Golgi membranes to the rod outer segment of the photoreceptor cell. Here we used synthetic peptides as competitive inhibitors of rhodopsin trafficking in the frog retinal cell-free system to delineate the potential regulatory sequence within the C terminus of rhodopsin and model the effects of severe retinitis pigmentosa alleles on rhodopsin sorting. The rhodopsin C-terminal sequence QVS(A)PA is highly conserved among different species. Peptides that correspond to the C terminus of bovine (amino acids 324-348) and frog (amino acids 330-354) rhodopsin inhibited post-Golgi trafficking by 50% and 60%, respectively, and arrested newly synthesized rhodopsin in the trans-Golgi network. Peptides corresponding to the cytoplasmic loops of rhodopsin and other control peptides had no effect. When three naturally occurring mutations: Q344ter (lacking the last five amino acids QVAPA), V345M, and P347S were introduced into the frog C-terminal peptide, the inhibitory activity of the peptides was no longer detectable. These observations suggest that the amino acids QVS(A)PA comprise a signal that is recognized by specific factors in the trans-Golgi network. A lack of recognition of this sequence, because of mutations in the last five amino acids causing autosomal dominant retinitis pigmentosa, most likely results in abnormal post-Golgi membrane formation and in an aberrant subcellular localization of rhodopsin.     

Optimizing Wireless LAN for Longwall Coal Mine Automation

A significant development in underground longwall coal mining automation has been achieved with the successful implementation of wireless LAN (WLAN) technology for communication on a longwall shearer. WIreless-FIdelity (Wi-Fi) was selected to meet the bandwidth requirements of the underground data network, and several configurations were installed on operating longwalls to evaluate their performance. Although these efforts demonstrated the feasibility of using WLAN technology in longwall operation, it was clear that new research and development was required in order to establish optimal full-face coverage. By undertaking an accurate characterization of the target environment, it has been possible to achieve great improvements in WLAN performance over a nominal Wi-Fi installation. This paper discusses the impact of Fresnel zone obstructions and multipath effects on radio frequency propagation and reports an optimal antenna and system configuration. Many of the lessons learned in the longwall case are immediately applicable to other underground mining operations, particularly wherever there is a high degree of obstruction from mining equipment     

Bioaccumulation factors for PCBs revisited

Bioaccumulation factors (BAFs) for individual polychlorinated biphenyl (PCB) congeners in Barents Sea and White Sea marine calanoid copepods were 1-3 orders of magnitude higher than BAFs in the same species in Canadian and Alaskan Arctic Ocean areas, and in freshwater plankton (Lake Ontario) reported from the mid- to early 1980s. The present study reviews variability in PCB BAFs from the North American Great Lakes and the Arctic Ocean, and discusses possible explanations for the large variation among different studies. BAFs are higher in recent arctic marine and Great Lakes studies than previously reported, and they are at least 10 times higher than those predicted from the octanol-water partition coefficient (KOW). If the recent high BAFs are realistic, it means that earlier reported BAFs are too low. This is likely due to earlier erroneously high quantification of water PCB concentrations, and it implies that bioaccumulation in zooplankton is more efficient than previously assumed. Evidence is presented supporting that also trophic transfer and biomagnification of PCBs in zooplankton leads to BAFs well above those predicted by simple equilibrium partitioning. Overall, miss-measurement of water PCB concentrations and biomagnification contribute significantly to variability in BAFs for PCBs within and among studies. This large variability of BAFs for PCBs in zooplankton illustrated in the present study is of importance for future assessments of potential new bioaccumulative chemicals that rely on measured BAFs, such as the European Union Registration, Evaluation and Authorization of Chemicals program (REACH).     

Silicon nitride Micromesh Bolometer Array for Submillimeter Astrophysics

We present the design and performance of a feedhorn-coupled bolometer array intended for a sensitive 350-mum photometer camera. Silicon nitride micromesh absorbers minimize the suspended mass and heat capacity of the bolometers. The temperature transducers, neutron-transmutation-doped Ge thermistors, are attached to the absorber with In bump bonds. Vapor-deposited electrical leads address the thermistors and determine the thermal conductance of the bolometers. The bolometer array demonstrates a dark noise-equivalent power of 2.9 x 10(-17) W/ radicalHz and a mean heat capacity of 1.3 pJ/K at 390 mK. We measure the optical efficiency of the bolometer and feedhorn to be 0.45-0.65 by comparing the response to blackbody calibration sources. The bolometer array demonstrates theoretical noise performance arising from the photon and the phonon and Johnson noise, with photon noise dominant under the design background conditions. We measure the ratio of total noise to photon noise to be 1.21 under an absorbed optical power of 2.4 pW. Excess noise is negligible for audio frequencies as low as 30 mHz. We summarize the trade-offs between bare and feedhorn-coupled detectors and discuss the estimated performance limits of micromesh bolometers. The bolometer array demonstrates the sensitivity required for photon noise-limited performance from a spaceborne, passively cooled telescope.     

Arctic marine ecosystem contamination.

The current state of knowledge of levels, spatial and temporal trends of contaminants in the Arctic marine ecosystem varies greatly among pollutants and among environmental compartments. Levels of polychlorinated biphenyls (PCBs), organochlorine (OC) pesticides and some heavy metals such as mercury and lead, in Arctic marine mammals and fish are relatively well documented because of the need for comparisons with biota in more polluted environments and interest in the contamination of native diets. Levels of heavy metals, alkanes, polyaromatic hydrocarbons (PAH) and OCs in the Arctic Ocean are comparable to uncontaminated ocean waters in the mid-latitudes. But concentrations of alpha- and gamma-hexachlorocyclohexane (HCHs) are higher in northern waters far removed from local sources, possibly because lower water temperature reduces transfer to the atmosphere. Bioaccumulation of OCs and heavy metals in Arctic marine food chains begins with epontic ice algae or phytoplankton in surface waters. Polychlorinated camphenes (PCC), PCBs, DDT- and chlordane-related compounds are the major OCs in marine fish, mammals and seabirds. Mean concentrations of most PCBs and OC pesticides in ringed seal (Phoca hispida) and polar bear (Ursus maritimus) populations in the Canadian Arctic are quite similar indicating a uniform geographic distribution of contamination, although alpha-HCH showed a distinct latitudinal gradient in bears due to higher levels in zones influenced by continental runoff. Ringed seals from Spitzbergen have higher levels of PCBs, total DDT and polychlorinated dioxins/furans (PCDD/PCDFs). In contrast to other OCs, PCDD/PCDFs in Canadian Arctic ringed seals and polar bears were higher in the east/central Arctic than at more southerly locations. Remarkably high cadmium levels are found in kidney and liver of narwhal (Monodons monoceros) from western Baffin Bay (mean of 63.5 micrograms g-1) and western Greenland waters (median of 39.5 micrograms g-1). Mercury concentrations in muscle of ringed seal and cetaceans frequently exceed 0.5 microgram g-1 especially in older animals. Cadmium concentrations in polar bear liver increased from west to east, while mercury levels were higher in ringed seals from the western Canadian Arctic, which suggests that natural sources of these metals predominate. Studies of temporal trends in OCs in ringed seals and seabirds in the Canadian Arctic indicate PCB and DDT levels declined significantly from the early 1970s to the 1980s. There is a lack of temporal trend data for other OC pesticides as well as for heavy metals and hydrocarbons.(ABSTRACT TRUNCATED AT 400 WORDS)     

3,4-Methylenedioxymethamphetamine Alters Left Ventricular Function and Activates Nuclear Factor-Kappa B (NF-κB) in a Time and Dose Dependent Manner

3,4-Methylenedioxymethamphetamine (MDMA) is an illicit psychoactive drug with cardiovascular effects that have not been fully described. In the current study, we observed the effects of acute MDMA on rabbit left ventricular function. We also observed the effects of MDMA on nuclear factor-kappa B (NF-κB) activity in cultured rat ventricular myocytes (H9c2). In the rabbit, MDMA (2 mg/kg) alone caused a significant increase in heart rate and a significant decrease in the duration of the cardiac cycle. Inhibition of nitric oxide synthase (NOS) by pretreatment with L-NAME (10 mg/kg) alone caused significant dysfunction in heart rate, systolic pressure, diastolic pressure, duration of relaxation, duration of cardiac cycle, and mean left ventricular pressure. Pretreatment with L-NAME followed by treatment with MDMA caused significant dysfunction in additional parameters that were not abnormal upon exposure to either compound in isolation: duration of contraction, inotropy, and pulse pressure. Exposure to 1.0 mM MDMA for 6 h or 2.0 μM MDMA for 12 h caused increased nuclear localization of NF-κB in cultured H9c2 cells. The current results suggest that MDMA is acutely detrimental to heart function and that an intact cardiovascular NOS system is important to help mitigate early sequelae in some functional parameters. The delayed timing of NF-κB activation suggests that this factor may be relevant to MDMA induced cardiomyopathy of later onset.     

A Multi‐Block Approach to Obtain Angle‐Resolved PIV Measurements of the Mean Flow and Turbulence Fields in a Stirred Vessel

Two‐dimensional Particle Image Velocimetry (PIV) measurements have been used to characterize the complex turbulent flow generated by a T/3 45° pitched‐blade down‐flow turbine, operated at Re ≈ 5 · 10⁴, in a fully turbulent stirred vessel. To maintain high spatial resolution when viewing the whole vessel, a multi‐block approach has been developed, which combines data from different fields of view into a composite flow map. Using 500 measurements of instantaneous u and v velocity fields, angle‐resolved mean velocity maps and turbulence properties, such as the RMS velocities and the turbulence kinetic energy, have been estimated near to the blade, as well as in the bulk of the vessel, at a spatial resolution of between 1 and 2 mm. Vorticity maps have also been calculated to help visualize the trailing vortex structures close to the impeller blades and integral length scales have been estimated from the two‐dimensional spatial auto‐correlation function. It is shown than the common assumption that the integral length scale is about half the blade width is an overestimate close to the impeller and an underestimate far from the impeller.     

Projection structure of frog rhodopsin in two crystal forms

Rhodopsin is the G protein-coupled receptor that upon light activation triggers the visual transduction cascade. Rod cell outer segment disc membranes were isolated from dark-adapted frog retinas and were extracted with Tween detergents to obtain two-dimensional rhodopsin crystals for electron crystallography. When Tween 80 was used, tubular structures with a p2 lattice (a = 32 A, b = 83 A, gamma = 91 degrees) were formed. The use of a Tween 80/Tween 20 mixture favored the formation of larger p22(1)2(1) lattices (a = 40 A, b = 146 A, gamma = 90 degrees). Micrographs from frozen hydrated frog rhodopsin crystals were processed, and projection structures to 7-A resolution for the p22(1)2(1) form and to 6-A resolution for the p2 form were calculated. The maps of frog rhodopsin in both crystal forms are very similar to the 9-A map obtained previously for bovine rhodopsin and show that the arrangement of the helices is the same. In a tentative topographic model, helices 4, 6, and 7 are nearly perpendicular to the plane of the membrane. In the higher-resolution projection maps of frog rhodopsin, helix 5 looks more tilted than it appeared previously. The quality of the two frog rhodopsin crystals suggests that they would be suitable to obtain a three-dimensional structure in which all helices would be resolved.