The cell cycle program of polypeptide labeling in Chlamydomonas reinhardtii

The cell cycle program of polypeptide labeling in syndhronous cultures of wild-type Chlamydomonas reinhardtii was analyzed by pulse-labeling cells with 35SO4 = or [3H]arginine at different cell cycle stages. Nearly 100 labeled membrane and soluble polypeptides were resolved and studied using one-dimensional sodium dodecyl sulfate (SDS)- polyacrylamide gel electrophoresis. The labeling experiments produced the following results. (a) Total 35SO4 = and [3H]arginine incorporation rates varied independently throughout the cell cycle. 35SO4 = incorporation was highest in the mid-light phase, while [3H]arginine incorporation peaked in the dark phase just before cell division. (b) The relative labeling rate for 20 of 100 polypeptides showed significant fluctuations (3-12 fold) during the cell cycle. The remaining polypeptides were labeled at a rate commensurate with total 35SO4 = or [3H]arginine incorporation. The polypeptides that showed significant fluctuations in relative labeling rates served as markers to identify cell cycle stages. (c) The effects of illumination conditions on the apparent cell cycle stage-specific labeling of polypeptides were tested. Shifting light-grown asynchronous cells to the dark had an immediate and pronounced effect on the pattern of polypeptide labeling, but shifting dark-phase syndhronous cells to the light had little effect. The apparent cell cycle variations in the labeling of ribulose 1,5-biphosphate (RUBP)-carboxylase were strongly influenced by illumination effects. (d) Pulse-chase experiments with light-grown asynchronous cells revealed little turnover or inter- conversion of labeled polypeptides within one cell generation, meaning that major polypeptides, whether labeled in a stage-specific manner or not, do not appear transiently in the cell cycle of actively dividing, light-grown cells. The cell cycle program of labeling was used to analyze effects of a temperature-sensitive cycle blocked (cb) mutant. A synchronous culture of ts10001 was shifted to restrictive temperature before its block point to prevent it from dividing. The mutant continued its cell cycle program of polypeptide labeling for over a cell generation, despite its inability to divide

Application of Suction-cup-attached VHF Transmitters to the Study of Beluga, Delphinapterus leucas, Surfacing Behavior in Cook Inlet, Alaska

Suction-cup-attached VHF radio transmittes were deployed on belugas, Delphinapterus leucas, in Cook Inlet, Alaska, in 1994 and 1995 to characterize the whales' surfacing behavior. Data from video recordings were also used to characterize behavior of undisturbed whales and whales actively pursued for tagging. Statistics for dive intervals (time between the midpoints of contiguous surfacings) and surfacing intevals (time at the surface per surfacing) were estimated. Operations took place on the tidal delta of the Susitna and Little Susitna Rivers. During the 2-yr study, eight whales were successfully tagged, five tags remained attached for >60 min, and data from these were used in the analyses. Mean dive interval was 24.1 sec (interwhale SD=6.4 sec, n=5). The mean surfacing interval, as determined from the duration of signals received from the radio transmitters, was 1.8 sec (SD=0.3 sec, n=125) for one of the whales. Videotaped behaviors were categorized as "head-lifts" or "slow-rolls." Belugas were more likely to head-lift than to slow-roll during vessel approaches and tagging attempts when compared to undisturbed whales. In undisturbed groups, surfacing intervals determined from video records were significantly different between head-lifting (average = 1.02 sect, SD=0.38 sed, n=28) and slow-rolling whales (average = 2.45 sec, SD=0.37 sec, n=106). Undisturbed juveniles exhibited shorter slow-roll surfacing intervals (average = 2.25 sec, SD=0.32 sec, n=36) than adults (average = 2.55 sec, SD=0.36 sec, n=70). We did not observe strong reactions by the belugas to the suction-cup tags. This tagging method shows promise for obtaining surfacing data for durations of several days

Expression of Heterologous OsDHAR Gene Improves Glutathione (GSH)-Dependent Antioxidant System and Maintenance of Cellular Redox Status in Synechococcus elongatus PCC 7942.

An excess of reactive oxygen species (ROS) can cause severe oxidative damage to cellular components in photosynthetic cells. Antioxidant systems, such as the glutathione (GSH) pools, regulate redox status in cells to guard against such damage. Dehydroascorbate reductase (DHAR, EC 1.8.5.1) catalyzes the glutathione-dependent reduction of oxidized ascorbate (dehydroascorbate) and contains a redox active site and glutathione binding-site. The DHAR gene is important in biological and abiotic stress responses involving reduction of the oxidative damage caused by ROS. In this study, transgenic Synechococcus elongatus PCC 7942 (TA) was constructed by cloning the Oryza sativa L. japonica DHAR (OsDHAR) gene controlled by an isopropyl β-D-1-thiogalactopyranoside (IPTG)-inducible promoter (Ptrc) into the cyanobacterium to study the functional activities of OsDHAR under oxidative stress caused by hydrogen peroxide exposure. OsDHAR expression increased the growth of S. elongatus PCC 7942 under oxidative stress by reducing the levels of hydroperoxides and malondialdehyde (MDA) and mitigating the loss of chlorophyll. DHAR and glutathione S-transferase activity were higher than in the wild-type S. elongatus PCC 7942 (WT). Additionally, overexpression of OsDHAR in S. elongatus PCC 7942 greatly increased the glutathione (GSH)/glutathione disulfide (GSSG) ratio in the presence or absence of hydrogen peroxide. These results strongly suggest that DHAR attenuates deleterious oxidative effects via the glutathione (GSH)-dependent antioxidant system in cyanobacterial cells. The expression of heterologous OsDHAR in S. elongatus PCC 7942 protected cells from oxidative damage through a GSH-dependent antioxidant system via GSH-dependent reactions at the redox active site and GSH binding site residues during oxidative stress

X-ray fluoresced high-Z (up to Z = 82) K-x-rays produced by LiNbO3 and LiTaO3 pyroelectric crystal electron accelerators

High-energy bremsstrahlung and K X-rays were used to produce nearly background-free K X-ray spectra of up to 87 keV (Pb) via X-ray fluorescence. The fluorescing radiation was produced by electron accelerators, consisting of heated and cooled cylindrical LiTaO3 and LiNbO3 crystals at mTorr pressures. The newly discovered process of gas amplification whereby the ambient gas pressure is optimized to maximize the electron energy was used to produce energetic electrons which when incident on a W/Bi target gave rise to a radiation field consisting of high-energy bremsstrahlung as well as W and Bi K X-rays. These photons were used to fluoresce Ta and Pb K X-rays.Comment: 6 pages, 6 figures, PD

A 10 GHz Quasi-Optical Grid Amplifier Using Integrated HBT Differential Pairs

We report the fabrication and testing of a 10 GHz grid amplifier utilizing sixteen GaAs chips each containing an HBT differential pair plus integral bias/feedback resistors. The overall amplifier consists of a 4x4 array of unit cells on an RT Duroid™ board having a relative permittivity of 2.2. Each unit cell consists of an emitter-coupled differential pair at the center, an input antenna which extends horizontally in both directions from the two base leads, an output antenna which extends vertically in both directions from the two collector leads, and high inductance bias lines. In operation, the active grid array is placed between a pair of crossed polarizers. The horizontally polarized input wave passes through the input polarizer and couples to the input leads. An amplified current then flows on the vertical leads, which radiate a vertically polarized amplified signal through the output polarizer. The polarizers serve dual functions, providing both input-output isolation as well as independent impedance matching for the input and output ports. The grid thus functions essentially as a free-space beam amplifier. Calculations indicate that output powers of several watts per square centimeter of grid area should be attainable with optimized structures

Seeking mechanisms for improved integration of biodiversity issues in regional natural resource management planning

This article reviews the literature on natural resource management (NRM) planning in Australia, with particular consideration given to exploring how regions might better integrate biodiversity conservation into catchment or regional planning in ways that lead to improved biodiversity conservation practice in the field. Many of the findings of the review are generic, affecting a range of NRM issues (including biodiversity conservation) and the NRM planning process itself, whilst other findings are specific to conservation of biodiversity. Factors affecting the integration of biodiversity include the organizational characteristics of the regional NRM body, clarity in the region of the responsibilities across the three tiers of government, effective participation of stakeholders, existence of detailed NRM plans that include sound biodiversity data and management principles, access to interpreted information, use of a mix of policy instruments capable of delivering biodiversity goals, and effective monitoring frameworks and tools to track the return on investment. there is considerable variability in the ways that NRM planning is practiced across Australia, at the enterprise, regional or catchment levels. However, an overarching issue is how well the planning caters for differences across space, time and human values and this article attempts to identify the considerations that impact on that requirement.<br /

Interfacial Reactions of Ozone with Surfactant Protein B in a Model Lung Surfactant System

Oxidative stresses from irritants such as hydrogen peroxide and ozone (O_3) can cause dysfunction of the pulmonary surfactant (PS) layer in the human lung, resulting in chronic diseases of the respiratory tract. For identification of structural changes of pulmonary surfactant protein B (SP-B) due to the heterogeneous reaction with O_3, field-induced droplet ionization (FIDI) mass spectrometry has been utilized. FIDI is a soft ionization method in which ions are extracted from the surface of microliter-volume droplets. We report structurally specific oxidative changes of SP-B_(1−25) (a shortened version of human SP-B) at the air−liquid interface. We also present studies of the interfacial oxidation of SP-B_(1−25) in a nonionizable 1-palmitoyl-2-oleoyl-sn-glycerol (POG) surfactant layer as a model PS system, where competitive oxidation of the two components is observed. Our results indicate that the heterogeneous reaction of SP-B_(1−25) at the interface is quite different from that in the solution phase. In comparison with the nearly complete homogeneous oxidation of SP-B_(1−25), only a subset of the amino acids known to react with ozone are oxidized by direct ozonolysis in the hydrophobic interfacial environment, both with and without the lipid surfactant layer. Combining these experimental observations with the results of molecular dynamics simulations provides an improved understanding of the interfacial structure and chemistry of a model lung surfactant system subjected to oxidative stress

Automated CFD shape optimization of stator blades for the PediaFlow pediatric ventricular assist device

PediaFlow is a miniature mixed-flow ventricular assist device for neonates and toddlers. PediaFlow has a fully magnetically levitated rotor which improves biocompatibility, but the increased length of the rotor creates a long annular passage where fluid energy is lost. Therefore, a set of helical stator blades was proposed immediately after the impeller stage to remove the swirling flow and recover the dynamic head as static pressure. Automated computational fluid dynamics (CFD) shape optimization of the stator blades was performed to maximize pressure recovery at the operating point of 1.5 LPM and 16,000 RPM. Additionally, the effect on hemolysis and thrombogenicity was assessed using numerical modeling. The optimization algorithm favored fewer blades of greater length over a larger number of short blades. The ratio of wrap angle to axial length emerged as a key constraint to ensure the viability of a design. The best design had 2 blades and generated 73 mmHg of pressure recovery in an isolated stage. When re-introduced to the CFD simulation of the complete flow path, the added stator stage increased the pump head by 46% and improved the pump efficiency from 21.9% to 25.7% at the selected operating point. Automated CFD shape optimization combined with in silico evaluation of hemocompatibility can be an effective tool for exploring design choices and informing early development process