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ATP-dependent Cl- uptake by plasma membrane vesicles from the rat brain.

Uptake of Cl- by plasma membrane vesicles from the rat brain was stimulated by ATP at 37 degrees C, but not by beta, gamma-methylene ATP or at 0 degrees C. The addition of Triton X-100 or sucrose to the incubation medium diminished the ATP-stimulated Cl- uptake, suggesting that Cl- was transported across the membranes into the intravesicular space. This ATP-stimulated Cl- uptake was not affected by 1 mM ouabain. 1 microM oligomycin, 0.1 mM gamma-aminobutyric acid or 0.1 mM picrotoxin. Thus, non-mitochondrial ATP-driven Cl- transport through a system other than Na, K-ATPase or Cl- channels occurs in neuronal plasma membrane vesicles.

Adenosine Triphosphatases↗

The role of extracellular calcium in exo- and endocytosis of synaptic vesicles at the frog motor nerve terminals.

In the present study we combined FM 1-43 imaging and electrophysiological recording of miniature end-plate currents (MEPCs) to determine the role of extracellular calcium in synaptic vesicle exo- and endocytosis at the frog motor nerve terminals. We replaced extracellular Ca2+ ions with other bivalent cations (Sr2+, Ba2+, Cd2+, Mg2+) or used a calcium-free solution and monitored fluorescent staining of the nerve terminals in the presence of caffeine, which promotes the release of Ca2+ from intracellular stores. Caffeine has induced FM1-43 internalization only in the presence of bivalent cations in the external solution. The exposure of the neuromuscular junction to caffeine in a calcium-free solution caused a reversible failure of FM 1-43 loading and an increase in the nerve terminal width. This effect of a calcium-free solution was not due to a decrease in exocytosis, because caffeine-induced FM1-43 unloading from the previously loaded nerve terminals, as well as a degree of the MEPCs frequency increase, was unchanged. We conclude that the presence of Ca2+ or other bivalent cations in extracellular space is necessary for endocytosis but not for exocytosis of synaptic vesicles, while transmitter release is promoted by efflux of Ca2+ from intracellular stores. The effect of extracellular Ca2+ on endocytosis might be driven by the non-specific interactions with membrane lipids.

Animals↗

Stimulus-driven and voluntary saccades are coded in different coordinate systems.

We make fast, "saccadic" eye movements to view our surroundings, "voluntary" saccades when saccade targets are deliberately selected, and "stimulus-driven" saccades when a target suddenly appears. Saccades of patients with spatial neglect have been studied to identify the coordinate systems guiding such behavior. However, previous reports disagree on whether neglect involves an eye-centered deficit of (delayed and hypometric) saccades specifically when performed in the direction opposite the brain lesion or not. We show that this inconsistency is due to independent mechanisms underlying voluntary and stimulus-driven saccades. We used a new experimental procedure comparing identical saccades performed either during an exploratory search task or a stimulus-driven task, both of which required similar cognitive functions (Figure 1). Only the patients' stimulus-driven saccades showed the eye-centered deficit. The same saccades were intact when voluntarily performed. However, here the patients showed a head-centered deficit; their saccades ignored the left part of space. In none of our control subjects with or without brain lesions did the neglect patients' pattern of deficits occur. The results argue that the brain flexibly uses a system of distinct but interrelated neural circuits for visual orienting to optimally encode its sensorimotor functions in multiple behavioral situations.

Brain Injuries↗

The jamming route to the glass state in weakly perturbed granular media.

It has been suggested that a common conceptual framework known as 'jamming' (refs 1 and 2) may be used to classify a wide variety of physical systems; these include granular media, colloidal suspensions and glass-forming liquids, all of which display a critical slowdown in their dynamics before a sudden transition to an amorphous rigid state. Decreasing the relevant control parameter (such as temperature, drive or inverse density) may cause geometrical constraints to build up progressively and thus restrict the accessible part of the system's phase space. In glass-forming liquids (thermal molecular systems), jamming is provided by the classical vitrification process of supercooling, characterized by a rapidly increasing and apparently diverging viscosity at sufficiently low temperatures. In driven (athermal) macroscopic systems, a similar slowdown has been predicted to occur, notably in sheared foam or vibrated granular media. Here we report experimental evidence for dynamic behaviour, qualitatively analogous to supercooling, in a driven granular system of macroscopic millimetre-size particles. The granular medium is perturbed by isolated tapping or continuous vibration, with the perturbation intensity serving as a control parameter. We observe the random deflection of an immersed torsion oscillator that moves each time the grains rearrange, like a 'thermometer' sensing the granular noise. We caution that our granular analogy to supercooling is based on similarities in the dynamical behaviour, rather than quantitative theory.

Journal Article↗

A satellite view of aerosols in the climate system.

Anthropogenic aerosols are intricately linked to the climate system and to the hydrologic cycle. The net effect of aerosols is to cool the climate system by reflecting sunlight. Depending on their composition, aerosols can also absorb sunlight in the atmosphere, further cooling the surface but warming the atmosphere in the process. These effects of aerosols on the temperature profile, along with the role of aerosols as cloud condensation nuclei, impact the hydrologic cycle, through changes in cloud cover, cloud properties and precipitation. Unravelling these feedbacks is particularly difficult because aerosols take a multitude of shapes and forms, ranging from desert dust to urban pollution, and because aerosol concentrations vary strongly over time and space. To accurately study aerosol distribution and composition therefore requires continuous observations from satellites, networks of ground-based instruments and dedicated field experiments. Increases in aerosol concentration and changes in their composition, driven by industrialization and an expanding population, may adversely affect the Earth's climate and water supply.

Aerosols↗

Localized adenovirus-mediated gene transfer into vascular smooth muscle in the hamster cheek pouch.

OBJECTIVE: Our purpose was to develop a method for adenovirus delivery to the hamster cheek pouch to experimentally target gene transfer in tissue used for microvascular studies. METHODS: Separate constructs were tested with transgenes for lacZ or green fluorescent protein (GFP) driven by three promoters: RSV, CMV, and SM22. With university approval, adenovirus was delivered in anesthetized (pentobarbital, 70 mg/kg) hamsters (n = 28) by using either a vascular systemic injection or tissue infiltration (interstitial space behind the pouch). During 3 days, animals receiving infiltration gained the expected weight, whereas those receiving vascular injection lost weight; no other behavior changes were noted. RESULTS: On day 3 postadenoviral delivery (infiltration), expression of lacZ (histology, beta-galactosidase) or GFP (fluorescence microscopy) was confirmed across the tissue (CMV and RSV promoters) and exclusively in vascular smooth muscle cells (specific SM22 promoter), without evidence of tissue inflammation. In vitro microvascular experiments verified normal responses in the cheek pouch of day 3 postadenoviral delivery animals. We tested local dilation to methacholine, adenosine, remote dilation to methacholine, adenosine, nitroprusside, and LM609 (alpha(v)beta3 integrin agonist), flow-dependent dilation, and flow recruitment. CONCLUSIONS: Thus, this method enables targeted, cell-specific gene transfer to one tissue important for microvascular studies, without significant systemic exposure and without adverse inflammation.

Adenoviridae↗

Tree water use and rainfall partitioning in a mature poplar-pasture system.

Traditionally, poplars (Populus) have been planted to control erosion on New Zealand's hill-slopes, because of their capacity to dry out and bind together the soil, by reducing effective rainfall and increasing evapotranspiration and soil strength. However, the effect of widely spaced poplars on the partitioning of soil water and rainfall has not been reported. This study determined rainfall partitioning for 18 mid-spring days in a mature P. deltoides (Bart. ex Marsh, Clone I78)-pasture association (37 stems per hectare, unevenly spaced at 16.4 +/- 0.4 m) and compared it with a traditional open pasture system in grazed areas of a hill environment. Tree transpiration was measured by the heat pulse technique. A time-driven mathematical model was used to set a zero offset, adjust anomalous values and describe simultaneous sap velocity time courses of trees. The model showed that daylight sap flow velocities can be represented with a nonlinear Beta function (R(2) > 0.98), and differences in the parameters representing the initiation, duration and conformation of the sap velocity can be tested statistically to discern tree transpiration differences during the day. Evapotranspiration was greater for the poplar-pasture association than for the open pasture (2.7-3.0 versus 2.2 mm day(-1)). The tree canopy alone contributed 0.92 mm day(-1) as transpiration and 1.37 mm day(-1) as interception, whereas evapotranspiration of the pasture understory was only 0.4-0.6 mm day(-1). Despite the higher water use of the poplar-pasture association, soil water in the 0-300 mm soil stratum was higher than, or similar to, that of the open pasture. Tree shading decreased evapotranspiration and pasture accumulation under the trees.

Journal Article↗

Role of Fluosol-DA 20% in prehospital resuscitation.

Fluosol-DA 20% (FDA-20) resuscitation has been proposed for prehospital therapy of hemorrhagic shock (HS). Acute HS (mean arterial pressure 60 mm Hg X 90 min, then 40 mm Hg X 30 min) in 24 splenectomized dogs was treated with 50 ml/kg of lactated Ringer's solution (RL) plus a volume equal to 105% of shed blood volume of FDA-20 (group 1), FDA-20 carrier Annex solution (group 2), or RL (group 3). Cardiopulmonary, hemopoietic, hemodynamic, and organ function parameters were measured preshock, at the end of shock, and at 30 and 60 min, and 24 h after resuscitation. FDA-20 produced effective volume expansion, oxygen delivery, and oxygen consumption. The FDA-20 appeared to affect organ function and cells adversely as reflected by a fall in red cell mass and platelet levels and by a rise in liver enzymes, BUN, and serum creatinine. Both the FDA-20 and Annex solution led to a reduction in serum proteins including serum albumin, serum globulin, immunoglobulin-G, and fibrinogen. These reductions are probably due to an oncotically driven factor which appears to maintain an optimal lymph to plasma oncotic ratio. The hydroxyethyl starch in the FDA-20 and the Annex solution, thus, would drive the plasma proteins into the interstitial space. The prolonged prothrombin times and the activated partial thromboplastin times after FDA-20 may be due, in part, to a reduction in the coagulation proteins, although these were not measured. Pending further studies designed to assess the effects of FDA-20 on possible cellular and multiple organ toxicity, clinical studies are premature and unwarranted.

Animals↗

Shear wave focusing for three-dimensional sonoelastography.

A new vibration scheme is shown to provide localized vibration fields for three-dimensional sonoelastography imaging. The theoretical vibration distributions of double strip loads vibrating normally to the surface of a semi-infinite elastic space are calculated. A localization or focusing of shear waves inbetween the double-strip loads is predicted. Experimentally, two parallel rigid rectangular cross-section bars are mounted on an electromagnetic shaker. Driven by the signal source, the bars vibrate against the surface of a tissue-mimicking phantom. The double-bar source is also used to propagate shear wave into an ex vivo prostate phantom with a 6 mm "tumor" in it. A combination of high frequencies (400-600 Hz) is used to drive the double-bar applicator. In the phantom experiments, a shear wave focal zone with higher vibration amplitude and uniformity predicted by the theory was confirmed. The position of the focal zone is controllable when adjusting the separation of the bars as the theory shows. When this vibration scheme was used in a prostate phantom experiment, high-resolution tumor images with clear boundaries are obtained. The parallel bar is an ideal applicator to create more uniform vibration within a controllable localized volume. The field has uniformity especially in the direction along the bars.

Culture Techniques↗

Contrast gain control and fine spatial discriminations.

The effect of contrast gain control mechanisms on discrimination between highly similar simple and complex stimuli is examined, with a focus on how discrimination accuracy changes as a function of the contrast of stimulus components. Two models of contrast gain control are evaluated. In both, the response of each pathway is attenuated by a factor determined by the total activity in a large pool of pathways. One model bases attenuation on the sum of linear filter responses within this pool; the other, based on Heeger's contrast energy-driven model [J. Neurophysiol. 70, 1985 (1993)], uses squared filter response. Predictions generated from the models are compared with data from experiments reported here and from the literature. Predictions are made for simple grafting stimuli of different sizes and for stimuli to which a second grafting component is added either as a second cue or as a mask. With one exception, predictions of the models agree closely with each other and with the data. The exception is a masking study that differentiates the models and supports the filter-driven model over the energy-driven model.

Contrast Sensitivity↗

Catabolism of 5-aminolevulinic acid to CO2 by rat liver mitochondria.

5-Aminolevulinic acid (ALA), the heme precursor accumulated in plasma and several organs of carriers of acute intermittent porphyria, hereditary tyrosinemia, and saturnism, was previously shown to yield reactive oxygen species upon metal-catalyzed aerobic oxidation and to cause the in vivo and in vitro impairment of rat liver mitochondrial functions. We have studied the uptake and catabolism of [5-14C]ALA to CO2 by isolated rat liver mitochondria (RLM) with the aim of determining whether possible ALA-driven oxidative injury to mitochondria can also occur into the matrix. Using silicone oil centrifugation of [5-14C]ALA-treated RLM, ALA was found to partition evenly into the intra- and extramatrix space of the mitochondrial preparations. The yield of evolved 14CO2 is very low (0.2%), responds to the concentration of added ADP, and is inhibited by malonate (75% at 2 mM), iproniazid (45% at 2 mM), beta-chloroalanine (36% at 1 mM), and aminooxyacetate (55% at 0.1 mM). With both iproniazid and aminooxyacetate, the percentage of inhibition is the same as that observed with the latter inhibitor alone. These data indicate that ALA decarboxylation by the Krebs cycle is a minor process and that it is initiated enzymically (transaminase) and not by metal-catalyzed ALA autoxidation.

Aminolevulinic Acid↗

Anomalous roughening of viscous fluid fronts in spontaneous imbibition.

We report experiments on spontaneous imbibition of a viscous fluid by a model porous medium in the absence of gravity. The average position of the interface satisfies Washburn's law. Scaling of the interface fluctuations suggests a dynamic exponent z approximately 3, indicative of global dynamics driven by capillary forces. The complete set of exponents clearly shows that interfaces are not self-affine, exhibiting distinct local and global scaling, both for time (beta = 0.64 +/- 0.02, beta(*) = 0.33 +/- 0.03) and space (alpha = 1.94 +/- 0.20, alpha(loc) = 0.94 +/- 0.10). These values are compatible with an intrinsic anomalous scaling scenario.

Journal Article↗

Overexpression of the zinc finger protein MZF1 inhibits hematopoietic development from embryonic stem cells: correlation with negative regulation of CD34 and c-myb promoter activity.

Zinc finger genes encode proteins that act as transcription factors. The myeloid zinc finger 1 (MZF1) gene encodes a zinc finger protein with two DNA-binding domains that recognize two distinct consensus sequences, is preferentially expressed in hematopoietic cells, and may be involved in the transcriptional regulation of hematopoiesis-specific genes. Reverse transcription-PCR analysis of human peripheral blood CD34+ cells cultured under lineage-restricted conditions demonstrated MZF1 expression during both myeloid and erythroid differentiation. Sequence analysis of the 5'-flanking region of the CD34 and c-myb genes, which are a marker of and a transcriptional factor required for hematopoietic proliferation and differentiation, respectively, revealed closely spaced MZF1 consensus binding sites found by electrophoretic mobility shift assays to interact with recombinant MZF1 protein. Transient or constitutive MZF1 expression in different cell types resulted in specific inhibition of chloramphenicol acetyltransferase activity driven by the CD34 or c-myb 5'-flanking region. To determine whether transcriptional modulation by MZF1 activity plays a role in hematopoietic differentiation, constructs containing the MZF1 cDNA under the control of different promoters were transfected into murine embryonic stem cells which, under defined in vitro culture conditions, generate colonies of multiple hematopoietic lineages. Constitutive MZF1 expression interfered with the ability of embryonic stem cells to undergo hematopoietic commitment and erythromyeloid colony formation and prevented the induced expression of CD34 and c-myb mRNAs during differentiation of these cells. These data indicate that MZF1 plays a critical role in hematopoiesis by modulating the expression of genes involved in this process.

Amino Acid Sequence↗

Reaction of the circulatory system to injury and regeneration.

The anatomy of the bone blood supply is well known. Techniques to measure bone blood flow are available, but no accurate method is available for clinical use. Ion movement from the capillary to the matrix is by diffusion, and efflux appears to be controlled by a concentration-dependent binding mechanism in the extravascular space of bone. The resistance vessels of bone are regulated by local, neural, and humoral factors. Fractures of long bone provoke a decrease in blood flow followed by a large increase in blood flow driven by metabolic demand. Fractures heal by coordinated formation of new bone, which is a combination of interstitial and surface remodeling. Age affects bone remodeling. Fixation of an experimental fracture may modify blood flow as well as bone remodeling. Fluid fluxes in bone may produce streaming potentials that in turn stimulate the osteoblasts to form new bone.

Animals↗

Short circuit of water vapor and polluted air to the global stratosphere by convective transport over the Tibetan Plateau.

During boreal summer, much of the water vapor and CO entering the global tropical stratosphere is transported over the Asian monsoon/Tibetan Plateau (TP) region. Studies have suggested that most of this transport is carried out either by tropical convection over the South Asian monsoon region or by extratropical convection over southern China. By using measurements from the newly available National Aeronautics and Space Administration Aura Microwave Limb Sounder, along with observations from the Aqua and Tropical Rainfall-Measuring Mission satellites, we establish that the TP provides the main pathway for cross-tropopause transport in this region. Tropospheric moist convection driven by elevated surface heating over the TP is deeper and detrains more water vapor, CO, and ice at the tropopause than over the monsoon area. Warmer tropopause temperatures and slower-falling, smaller cirrus cloud particles in less saturated ambient air at the tropopause also allow more water vapor to travel into the lower stratosphere over the TP, effectively short-circuiting the slower ascent of water vapor across the cold tropical tropopause over the monsoon area. Air that is high in water vapor and CO over the Asian monsoon/TP region enters the lower stratosphere primarily over the TP, and it is then transported toward the Asian monsoon area and disperses into the large-scale upward motion of the global stratospheric circulation. Thus, hydration of the global stratosphere could be especially sensitive to changes of convection over the TP.

Air↗

Human body-segment tilts induced by galvanic stimulation: a vestibularly driven balance protection mechanism.

1. We have studied the effects of changes in posture on the motor response to galvanic vestibular stimulation (GVS). The purpose of the experiments was to investigate whether the function of the GVS-evoked response is to stabilize the body or the head in space. Subjects faced forwards with eyes closed standing with various stance widths and sitting. In all cases the GVS-evoked response consisted of a sway of the body towards the anodal ear. 2. In the first set of experiments the response was measured from changes in (i) electromyographic activity of hip and ankle muscles, (ii) the lateral ground reaction force, and (iii) lateral motion of the body at the level of the neck (C7). For all measurements the response became smaller as the feet were placed further apart. 3. In the second set of experiments we measured the GVS-evoked tilts of the head, torso and pelvis. The basic response consisted of a tilt in space (anodal ear down) of all three segments. The head tilted more than the trunk and the trunk tilted more than the pelvis producing a leaning and bending of the body towards the anodal ear. This change in posture was sustained for the duration of the stimulus. 4. The tilt of all three segments was reduced by increasing the stance width. This was due to a reduction in evoked tilt of the pelvis, the bending of the upper body remaining relatively unchanged. Changing from a standing to a sitting posture produced additional reductions in tilt by reducing the degree of upper body bending. 5. The results indicate that the response is organized to stabilize the body rather than the head in space. We suggest that GVS produces a vestibular input akin to that experienced on an inclined support surface and that the function of the response is to counter any threat to balance by keeping the centre of mass of the body within safe limits.

Electric Stimulation↗

Agonistic activity and charge driven interaction potentiality of PAF derivatives.

Taking into account the contribution of the C2 substituent to the interaction ability of PAF derivatives, an investigation has been undertaken on: i) the role of the presence of two unitary charges of opposite sign in the molecule; ii) the role of the spacing of the same charged moieties. For this purpose, the theoretical analysis of model compounds for newly synthesized and already known compounds has been performed by means of computer-aided modelling, molecular mechanics and semiempirical quantum chemical computational methods. The combination of the results with the available pharmacological data allows a discussion about the stereoelectronic features required for agonist interaction at the PAF receptor.

Animals↗

Direction-selective responses of units in the dorsal terminal nucleus of cat following intravitreal injections of bicuculline.

Extracellular recordings from single units in the dorsal terminal nucleus (DTN) of the cat accessory optic system (AOS) were made before and after intravitreal injections of the GABA antagonist bicuculline methiodide (BMI). Direction-selective responses of DTN cells elicited through the contralateral, injected eye were abolished 7-12 h following the injection. For the concentrations tested, direction-selective responses through the contralateral (injected) eye did not recover within 26 h. Direction-selective responses through stimulation of the ipsilateral (uninjected) eye were also dramatically depressed for 1-9 h after contralateral eye injections. However, direction-selective responses through the ipsilateral eye eventually returned and were often more vigorous in the final stages. BMI injections into the ipsilateral eye failed to block direction-selective responses through the ipsilateral eye. The effects of intravitreal BMI on contralateral eye responses imply that DTN units receive input from direction-selective retinal ganglion cells. In addition, these results suggest that direction-selective input to the DTN from the visual cortex is independent of the retinal pathway. Using pharmacological methods described here, for the first time direction-selective responses of AOS units driven through the ipsilateral eye can be experimentally isolated.

Animals↗