Differential expression of dystrophin isoforms and utrophin during dibutyryl-cAMP-induced morphological differentiation of rat brain astrocytes

We have identified isoforms of dystrophin and utrophin, a dystrophin homologue, expressed in astrocytes and examined their expression patterns during dibutyryl-cAMP (dBcAMP)-induced morphological differentiation of astrocytes. Immunoblot and immunocytochemical analyses showed that full-length-type dystrophin (427 kDa), utrophin (395 kDa), and Dp71 (75 kDa), a small-type dystrophin isoform, were coexpressed in cultured nondifferentiated rat brain astrocytes and were found to be located in the cell membrane. During morphological differentiation of the astrocytes induced by 1 mM dBcAMP, the amount of Dp71 markedly increased, whereas that of dystrophin and utrophin decreased. Northern blot analyses revealed that dBcAMP regulates the mRNA levels of Dp71 and dystrophin but not that of utrophin. dBcAMP slightly increased the amount of the β-dystroglycan responsible for anchoring dystrophin isoforms and utrophin to the cell membrane. Immunocytochemical analyses showed that most utrophin was observed in the cytoplasmic area during astrocyte differentiation, whereas Dp71 was found along the cell membrane of the differentiated astrocytes. These findings suggest that most of the dystrophin/utrophin-dystroglycan complex on cell membrane in cultured astrocytes was replaced by the Dp71-dystroglycan complex during morphological differentiation. The cell biological roles of Dp71 are discussed.

Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex

Cortical blood flow at the level of individual capillaries and the coupling of neuronal activity to flow in capillaries are fundamental aspects of homeostasis in the normal and the diseased brain. To probe the dynamics of blood flow at this level, we used two-photon laser scanning microscopy to image the motion of red blood cells (RBCs) in individual capillaries that lie as far as 600 μm below the pia mater of primary somatosensory cortex in rat; this depth encompassed the cortical layers with the highest density of neurons and capillaries. We observed that the flow was quite variable and exhibited temporal fluctuations around 0.1 Hz, as well as prolonged stalls and occasional reversals of direction. On average, the speed and flux (cells per unit time) of RBCs covaried linearly at low values of flux, with a linear density of ≈70 cells per mm, followed by a tendency for the speed to plateau at high values of flux. Thus, both the average velocity and density of RBCs are greater at high values of flux than at low values. Time-locked changes in flow, localized to the appropriate anatomical region of somatosensory cortex, were observed in response to stimulation of either multiple vibrissae or the hindlimb. Although we were able to detect stimulus-induced changes in the flux and speed of RBCs in some single trials, the amplitude of the stimulus-evoked changes in flow were largely masked by basal fluctuations. On average, the flux and the speed of RBCs increased transiently on stimulation, although the linear density of RBCs decreased slightly. These findings are consistent with a stimulus-induced decrease in capillary resistance to flow.

Increased histone H1 phosphorylation and relaxed chromatin structure in Rb-deficient fibroblasts.

Fibroblasts derived from embryos homozygous for a disruption of the retinoblastoma gene (Rb) exhibit a shorter G1 than their wild-type counterparts, apparently due to highly elevated levels of cyclin E protein and deregulated cyclin-dependent kinase 2 (CDK2) activity. Here we demonstrate that the Rb-/- fibroblasts display higher levels of phosphorylated H1 throughout G1 with the maximum being 10-fold higher than that of the Rb+/+ fibroblasts. This profile of intracellular H1 phosphorylation corresponds with deregulated CDK2 activity observed in in vitro assays, suggesting that CDK2 may be directly responsible for the in vivo phosphorylation of H1. H1 phosphorylation has been proposed to lead to a relaxation of chromatin structure due to a decreased affinity of this protein for chromatin after phosphorylation. In accord with this, chromatin from the Rb-/- cells is more susceptible to micrococcal nuclease digestion than that from Rb+/+ fibroblasts. Increased H1 phosphorylation and relaxed chromatin structure have also been observed in cells expressing several oncogenes, suggesting a common mechanism in oncogene and tumor suppressor gene function.

FM1-43 dye ultrastructural localization in and release from frog motor nerve terminals.

Previous work has shown that the fluorescent styryl dye FM1-43 stains nerve terminals in an activity-dependent fashion. This dye appears to label the membranes of recycled synaptic vesicles by being trapped during endocytosis. Stained terminals can subsequently be destained by repeating nerve stimulation in the absence of dye; the destaining evidently reflects escape of dye into the bathing medium from membranes of exocytosing synaptic vesicles. In the present study we tested two key aspects of this interpretation of FM1-43 behavior, namely: (i) that the dye is localized in synaptic vesicles, and (ii) that it is actually released into the bathing medium during destaining. To accomplish this, we first photolyzed the internalized dye in the presence of diaminobenzidine. This created an electron-dense reaction product that could be visualized in the electron microscope. Reaction product was confined to synaptic vesicles, as predicted. Second, using spectrofluorometry, we quantified the release of dye liberated into the medium from tubocurarine-treated nerve-muscle preparations. Nerve stimulation increased the amount of FM1-43 released, and we estimate that normally a stained synaptic vesicle contains a few hundred molecules of the dye. The key to the successful detection of released FM1-43 was to add the micelle-forming detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), which increased FM1-43 quantum yield by more than two orders of magnitude.

cAMP- and rapamycin-sensitive regulation of the association of eukaryotic initiation factor 4E and the translational regulator PHAS-I in aortic smooth muscle cells.

Incubating rat aortic smooth muscle cells with either platelet-derived growth factor BB (PDGF) or insulin-like growth factor I (IGF-I) increased the phosphorylation of PHAS-I, an inhibitor of the mRNA cap binding protein, eukaryotic initiation factor (eIF) 4E. Phosphorylation of PHAS-I promoted dissociation of the PHAS-I-eIF-4E complex, an effect that could partly explain the stimulation of protein synthesis by the two growth factors. Increasing cAMP with forskolin decreased PHAS-I phosphorylation and markedly increased the amount of eIF-4E bound to PHAS-I, effects consistent with an action of cAMP to inhibit protein synthesis. Both PDGF and IGF-I activated p70S6K, but only PDGF increased mitogen-activated protein kinase activity. Forskolin decreased by 50% the effect of PDGF on increasing p70S6K, and forskolin abolished the effect of IGF-I on the kinase. The effects of PDGF and IGF-I on increasing PHAS-I phosphorylation, on dissociating the PHAS-I-eIF-4E complex, and on increasing p70S6K were abolished by rapamycin. The results indicate that IGF-I and PDGF increase PHAS-I phosphorylation in smooth muscle cells by the same rapamycin-sensitive pathway that leads to activation of p70S6K.

Response of Catch-Per-Unit-Effort to Spatial and Environmental Factors: an Exploratory Data Analysis Using Generalized Additive Models

Understanding spatial distributions and how environmental conditions influence catch-per-unit-effort (CPUE) is important for increased fishing efficiency and sustainable fisheries management. This study investigated the relationship between CPUE, spatial factors, temperature, and depth using generalized additive models. Combinations of factors, and not one single factor, were frequently included in the best model. Parameters which best described CPUE varied by geographic region. The amount of variance, or deviance, explained by the best models ranged from a low of 29% (halibut, Charlotte region) to a high of 94% (sablefish, Charlotte region). Depth, latitude, and longitude influenced most species in several regions. On the broad geographic scale, depth was associated with CPUE for every species, except dogfish. Latitude and longitude influenced most species, except halibut (Areas 4 A/D), sablefish, and cod. Temperature was important for describing distributions of halibut in Alaska, arrowtooth flounder in British Columbia, dogfish, Alaska skate, and Aleutian skate. The species-habitat relationships revealed in this study can be used to create improved fishing and management strategies.

Beyond the H2/CO2 upper bound: one-step crystallization and separation of nano-sized ZIF-11 by centrifugation and its application in mixed matrix membranes

The synthesis of nano-sized ZIF-11 with an average size of 36 ± 6 nm is reported. This material has been named nano-zeolitic imidazolate framework-11 (nZIF-11). It has the same chemical composition and thermal stability and analogous H2 and CO2 adsorption properties to the conventional microcrystalline ZIF-11 (i.e. 1.9 ± 0.9 μm). nZIF-11 has been obtained following the centrifugation route, typically used for solid separation, as a fast new technique (pioneering for MOFs) for obtaining nanomaterials where the temperature, time and rotation speed can easily be controlled. Compared to the traditional synthesis consisting of stirring + separation, the reaction time was lowered from several hours to a few minutes when using this centrifugation synthesis technique. Employing the same reaction time (2, 5 or 10 min), micro-sized ZIF-11 was obtained using the traditional synthesis while nano-scale ZIF-11 was achieved only by using centrifugation synthesis. The small particle size obtained for nZIF-11 allowed the use of the wet MOF sample as a colloidal suspension stable in chloroform. This helped to prepare mixed matrix membranes (MMMs) by direct addition of the membrane polymer (polyimide Matrimid®) to the colloidal suspension, avoiding particle agglomeration resulting from drying. The MMMs were tested for H2/CO2 separation, improving the pure polymer membrane performance, with permeation values of 95.9 Barrer of H2 and a H2/CO2 separation selectivity of 4.4 at 35 °C. When measured at 200 °C, these values increased to 535 Barrer and 9.1.

Water polo throwing speed and body composition: an analysis by playing positions and opposition level

The aim of this study was to develop an anthropometric profile on highly skilled male water polo players by specific playing positions. Also, to identify significant relationships between these features an overhead throwing speed in highly skilled male Water Polo players by specific playing positions. Methods: A total of 94 male water polo players (24.5±5.3 yrs) who were playing in the Spanish King´s cup were studied. Subjects were grouped according to their specific playing positions: 15 goalkeepers, 45 offensive wings, 20 center backs and 14 center forwards. Anthropometric assessment was made following ISAK protocols. Hand grip and throwing speed in several situations were also assessed. A one-way analysis of variance (ANOVA) was used to determine if significant differences existed among the four playing positions. Pearson product-moment correlation coefficients (r) were used to determine the relationships of all anthropometric measures with throwing speed and hand grip. The total player’s somatotype was endomorphic-mesomorphic (2.9–5.8–2.3). Center forwards exhibit important anthropometric differences compared with the other specific playing positions in elite male water polo players, but no differences were found in throwing speed by specific playing positions in each throwing conditions. Moreover, a higher number of relationships between anthropometric and throwing speed were found in wings and also in center backs but no relationships were found in center forwards. The data reflects the importance of muscle mass and upper body in the throwing skill. Coaches can use this information in order to select players for the different specific positions.

LCD vs. E-ink: An Analysis of the Reading Behavior

Electronic books (e-book) are an interesting option compared to classic paper books. Most e-reading devices of the first generation were based on e-ink technology. With the appearance of the Apple iPad on the market, TFT-LCDs became important in the field of e-reading. Both technologies have advantages and disadvantages but the question remains whether one or the other technology is better for reading. In the present study we analyzed and compared reading behavior when reading on e-inkreader (e-ink displays) and on tablets (TFT-LCDs) as measured by eye-tracking. The results suggest that the reading behavior on tablets is indeed very similar to the reading behavior on e-ink-reader. Participants showed no difference in fixation duration. Significant differences in reading speed and in the proportion of regressive saccades suggest that tablets, under special artificial light conditions, may even provide better legibility.

High-speed UDP Data Transmission with Multithreading and Automatic Resource Allocation

In this paper a utilization of the high data-rates channels by threading of sending and receiving is studied. As a communication technology evolves the higher speeds are used more and more in various applications. But generating traffic with Gbps data-rates also brings some complications. Especially if UDP protocol is used and it is necessary to avoid packet fragmentation, for example for high-speed reliable transport protocols based on UDP. For such situation the Ethernet network packet size has to correspond to standard 1500 bytes MTU[1], which is widely used in the Internet. System may not has enough capacity to send messages with necessary rate in a single-threaded mode. A possible solution is to use more threads. It can be efficient on widespread multicore systems. Also the fact that in real network non-constant data flow can be expected brings another object of study –- an automatic adaptation to the traffic which is changing during runtime. Cases investigated in this paper include adjusting number of threads to a given speed and keeping speed on a given rate when CPU gets heavily loaded by other processes while sending data.

The bibliographer's manual of English literature containing an account of rare, curious, and useful books, published in or relating to Great Britain and Ireland, from the invention of printing; with bibliographical and critical notices, collations of the rarer articles, and the prices at which they have been sold.

Vol. 6 has special title: Appendix to the Bibliographer's manual of English literature. Containing an account of books issued by literary and scientific societies and printing clubs; books printed at private presses; privately printed series; and the principal literary and scientific serials. Comp. by Henry G. Bohn ...

Two NextGen Air Safety Tools: An ADS-B Equipped UAV and a Wake Turbulence Estimator

Thesis (Master's)--University of Washington, 2016-06

Effects of boron foliar applications on vegetative and reproductive growth of sunflower

Foliar application may be used to supply boron (B) to a crop when B demands are higher than can be supplied via the soil. While B foliar sprays have been used to correct B deficiency in sunflower (Helianthus annuus L.) in the field, no studies have determined the amount of B taken up by sunflower plant parts via foliar application. A study was conducted in which sunflower plants were grown at constant B concentration in nutrient solution with adequate B (46 mum) or with limited B supply (0.24, 0.40 and 1.72 mum) using Amberlite IRA-743 resin to control B supply. At the late vegetative stage of growth (25 and 35 d after transplanting), two foliar sprays were applied of soluble sodium tetraborate (20.8 % B) each at 0, 28, 65, 120 and 1200 mm (each spray equivalent to 0, 0.03, 0.07, 0.13 and 1.3 kg B ha(-1) in 100 L water ha(-1)). The highest rate of B foliar fertilization resulted in leaf burn but had no other evident detrimental effect on plant growth. Under B-deficient conditions, B foliar application increased the vegetative and reproductive dry mass of plants. Foliar application of 28-1200 mm B increased the total dry mass of the most B-deficient plants by more than three-fold and that of plants grown initially with 1.72 mum B in solution by 37-49 %. In this latter treatment, the dry mass of the capitulum was similar to that achieved under control conditions, but in no instance was total plant dry mass similar to that of the control. All B foliar spray rates increased the B concentration in various parts of the plant tops, including those that developed after the sprays were applied, but the B concentration in the roots was not increased by B foliar application. The B concentration in the capitulum of the plants sprayed at the highest rate was between 37 and 93 % of that in the control plants. This study showed that B foliar application was of benefit to B-deficient sunflower plants, increasing the B status of plant tops, including that of the capitulum which developed after the B sprays were applied. (C) 2003 Annals of Botany Company.

Tennis, incidence of URTI and salivary IgA

Tennis played at an elite level requires intensive training characterized by repeated bouts of brief intermittent high intensity exercise over relatively long periods of time (1 - 3 h or more). Competition can place additional stress on players. The purpose of this study was to investigate the temporal association between specific components of tennis training and competition, the incidence of upper respiratory tract infections (URT1), and salivary IgA, in a cohort of seventeen elite female tennis players. Timed, whole unstimulated saliva samples were collected before and after selected 1-h training sessions at 2 weekly intervals, over 12 weeks. Salivary IgA concentration was measured by ELISA and IgA secretion rate calculated (mug IgA x ml(-1) x ml saliva x min(-1)). Players reported URTI symptoms and recorded training and competition in daily logs. Data analysis showed that higher incidence of URTI was significantly associated with increased training duration and load, and competition level, on a weekly basis. Salivary IgA secretion rate (S-IgA) dropped significantly after 1 hour of tennis play. Over the 12-week period, pre-exercise salivary IgA concentration and secretion rate were directly associated with the amount of training undertaken during the previous day and week (p < 0.05). However, the decline in S-IgA after 1 h of intense tennis play was also positively related to the duration and load of training undertaken during the previous day and week (p < 0.05). Although exercise-induced suppression of salivary IgA may be a risk factor, it could not accurately predict the occurrence of URTI in this cohort of athletes.

Comparative atmospheric corrosion of primary and cold rolled copper in Australia

Atmospheric corrosion tests, according to ASTM G50, have been carried out in Queensland, Australia, at three different sites representing three different environmental conditions. A range of materials including primary copper (electrosheet) and electrolytic tough pitch (traditional cold rolled) copper have been exposed. Data is available for five exposure periods over a three year time span. X-Ray Diffraction has been used to determine the composition of the corrosion products. Corrosion rates have been determined for each material at each of the exposure sites and are compared with corrosion rates obtained from other long term atmospheric corrosion test programs. Primary copper sheet (electrosheet) behaves like traditionally produced cold rolled copper (C11000) sheet but with an increased corrosion rate. This difference between the rolled copper samples and the primary copper samples is probably due to a combination of factors related to the difference in crystallographic texture of the underlying copper, the morphology and texture of the cuprite layer, the surface roughness of the sheets, and the differences in mass. These factors combine together to provide an increased oxidation rate and TOW for the electrosheet material and which is significantly higher at the more tropical sites. For a sulfate environment (Urban) the initial corrosion product is cuprite with posnjakite and brochantite also occurring at longer exposures. Posnjakite is either washed away or converted to brochantite during further exposure. The amount of brochantite increases with exposure time and forms the blue-green patina layer. For a chloride environment (Marine) the initial corrosion product is cuprite with atacamite also occurring at longer exposures.