scholarly journals 54Mn2+ as a tracer of the polymerization of actin. Intermediate oligomers condense to give F-actin

1984 ◽  
Vol 223 (3) ◽  
pp. 571-576 ◽  
Author(s):  
E Grazi
Keyword(s):  
Steady State ◽  
Association Constant ◽  
Time Course ◽  
Protein Analysis ◽  
Effective Mechanism ◽  
Experimental Conditions ◽  
Initial Concentration ◽  
Direct Condensation

Mg2+, at submicromolar concentrations, is needed for the nucleation of actin [Maruyama (1981) J. Biol. Chem. 256, 1060-1062]. I show here that Mn2+ fulfils the same function. It binds to oligomers present in the ATP-G-actin solutions with a ratio of 2-3 Mn2+ ions per 100 actin monomers and with an association constant of 0.66 × 10(10) M-1 at pH 8.2 at 25 degrees C. The time course of the binding of Mn2+ to polymerizing actin is not affected by the initial concentration of the protein. Analysis of the distribution of the binding shows that, both in the large oligomeric species and in the polymers, 1 Mn2+ ion is bound for every 14-25 actin monomers, whereas in the smaller oligomeric species 1 Mn2+ ion is bound for every 4 actin monomers. The proposal is made that Mn2+ stabilizes actin nuclei and decreases the concentration of the monomers at the steady state. It is also proposed that, at least in some experimental conditions, the direct condensation of oligomers of intermediate length is an effective mechanism of F-actin formation.

scholarly journals Genome-wide dynamics of RNA synthesis, processing and degradation without RNA metabolic labeling

2019 ◽  
Author(s):  
Mattia Furlan ◽  
Eugenia Galeota ◽  
Nunzio Del Gaudio ◽  
Erik Dassi ◽  
Michele Caselle ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Steady State ◽  
Time Course ◽  
Rna Synthesis ◽  
Computational Method ◽  
Metabolic Labeling ◽  
Experimental Conditions ◽  
Rna Dynamics ◽  
Kinetic Rates ◽  
Post Transcriptional Regulation

AbstractThe kinetic rates of RNA synthesis, processing and degradation determine the dynamics of transcriptional regulation by governing both the abundance and the responsiveness to modulations of premature and mature RNA species. The study of RNA dynamics is largely based on the integrative analysis of total and nascent transcription, with the latter being quantified through RNA metabolic labeling. We describe here INSPEcT-, a computational method based on mathematical modeling of intronic and exonic expression, able to derive the dynamics of transcription from steady state or time course profiling of just total RNA, without requiring any information on nascent transcripts. Our approach closely recapitulates the kinetic rates obtained through RNA metabolic labeling, improves the ability to detect changes in transcripts half-lives, reduces the cost and complexity of the experiments, and can be adopted to study experimental conditions where nascent transcription cannot be readily profiled. Finally, we applied INSPEcT- to the characterization of post-transcriptional regulation landscapes in dozens of physiological and disease conditions. This approach was included in the INSPEcT Bioconductor package, which can now unveil RNA dynamics from steady state or time course data, with or without the profiling of nascent RNA.

A Simple Method for Estimation of Lipoxygenase and Cyclo-Oxygenase Pathways in Human Platelets - the Use of Thiobarbituric Acid Reaction

1980 ◽  
Vol 44 (02) ◽  
pp. 111-114 ◽  
Author(s):  
Hiroshi Takayama ◽  
Minoru Okuma ◽  
Haruto Uchino
Keyword(s):  
Time Course ◽  
Normal Values ◽  
Human Platelet ◽  
Thiobarbituric Acid ◽  
Human Platelets ◽  
Optimal Conditions ◽  
Experimental Conditions ◽  
Simple Method ◽  
Acid Reaction ◽  
Optimal Ph

SummaryTo develop a simple method for estimation of platelet lipoxygenase (PLO) and cyclo-oxygenase (PCO) pathways, the arachidonic acid (AA) metabolism of human platelet was investigated under various experimental conditions by the use of the thiobarbituric acid (TBA) reaction and a radioisotope technique. A TBA-reactive substance different from malondialdehyde (MDA) via PCO pathway was detected and shown to be derived from the PLO pathway. Since the optimal pH and time course of its formation were different from those of MDA formation via PCO pathway, PLO and PCO pathways were estimated by quantitating the TBA-reactive substances produced by the incubation of AA either with aspirin-treated platelets or with untreated ones, respectively, each under optimal conditions. Normal values expressed in terms of nmol MDA/108 platelets were 1.17±0.34 (M±SD, n = 31) and 0.79±0.15 (n = 31) for PLO and PCO pathways, respectively.

scholarly journals The steady-state visual evoked potential (SSVEP) reflects the activation of cortical object representations: evidence from semantic stimulus repetition

2020 ◽  
Author(s):  
Elise L. Radtke ◽  
Ulla Martens ◽  
Thomas Gruber
Keyword(s):  
Steady State ◽  
Object Representation ◽  
Sensory Information ◽  
Familiar Object ◽  
Object Representations ◽  
Stimulus Repetition ◽  
Experimental Conditions ◽  
Repetition Suppression ◽  
Task Irrelevant ◽  
Visual Evoked

AbstractWe applied high-density EEG to examine steady-state visual evoked potentials (SSVEPs) during a perceptual/semantic stimulus repetition design. SSVEPs are evoked oscillatory cortical responses at the same frequency as visual stimuli flickered at this frequency. In repetition designs, stimuli are presented twice with the repetition being task irrelevant. The cortical processing of the second stimulus is commonly characterized by decreased neuronal activity (repetition suppression). The behavioral consequences of stimulus repetition were examined in a companion reaction time pre-study using the same experimental design as the EEG study. During the first presentation of a stimulus, we confronted participants with drawings of familiar object images or object words, respectively. The second stimulus was either a repetition of the same object image (perceptual repetition; PR) or an image depicting the word presented during the first presentation (semantic repetition; SR)—all flickered at 15 Hz to elicit SSVEPs. The behavioral study revealed priming effects in both experimental conditions (PR and SR). In the EEG, PR was associated with repetition suppression of SSVEP amplitudes at left occipital and repetition enhancement at left temporal electrodes. In contrast, SR was associated with SSVEP suppression at left occipital and central electrodes originating in bilateral postcentral and occipital gyri, right middle frontal and right temporal gyrus. The conclusion of the presented study is twofold. First, SSVEP amplitudes do not only index perceptual aspects of incoming sensory information but also semantic aspects of cortical object representation. Second, our electrophysiological findings can be interpreted as neuronal underpinnings of perceptual and semantic priming.

Inward rectification in rat nucleus accumbens neurons

1989 ◽  
Vol 62 (6) ◽  
pp. 1280-1286 ◽  
Author(s):  
N. Uchimura ◽  
E. Cherubini ◽  
R. A. North
Keyword(s):  
Steady State ◽  
Nucleus Accumbens ◽  
Time Course ◽  
Potassium Conductance ◽  
Resting Potential ◽  
Inward Rectification ◽  
Resting Membrane Potential ◽  
Potential Range ◽  
Inward Current ◽  
Rat Nucleus Accumbens

1. Intracellular recordings were made from neurons in slices cut from the rat nucleus accumbens septi. Membrane currents were measured with a single-electrode voltage-clamp amplifier in the potential range -50 to -140 mV. 2. In control conditions (2.5 mM potassium), the resting membrane potential of the neurons was -83.4 +/- 1.1 (SE) mV (n = 157). Steady state membrane conductance was voltage dependent, being 34.8 +/- 1.7 nS (n = 25) at -100 mV and 8.0 +/- 0.7 nS (n = 25) at -60 mV. 3. Barium (1 microM) markedly reduced the inward rectification and caused a small inward current (40.6 +/- 8.7 pA, n = 8) at the resting potential. These effects became larger with higher barium concentrations, and, in 100 microM barium, the current-voltage relation was straight. 4. The block of the inward current by barium (at -130 mV) occurred with an exponential time course; the time constant was approximately 1 s at 1 microM barium and less than 90 ms with 100 microM. Strontium had effects similar to those of barium, but 1000-fold higher concentrations were required. Cesium chloride (2 mM) and rubidium chloride (2 mM) also blocked the inward rectification; their action reached steady state within 50 ms. 5. It is concluded that the nucleus accumbens neurons have a potassium conductance with many features of a typical inward rectifier and that this contributes to the potassium conductance at the resting potential.

scholarly journals Participation of plasma membrane proteins in the formation of tight junction by cultured epithelial cells

1983 ◽  
Vol 96 (3) ◽  
pp. 693-702 ◽  
Author(s):  
EB Griepp ◽  
WJ Dolan ◽  
ES Robbins ◽  
DD Sabatini
Keyword(s):  
Protein Synthesis ◽  
Steady State ◽  
Tight Junction ◽  
Messenger Rna ◽  
Freeze Fracture ◽  
Epithelial Cell Line ◽  
Experimental Conditions ◽  
Tight Junction Formation ◽  
Junction Formation ◽  
Spinner Culture

Measurements of the transepithelial electrical resistance correlated with freeze-fracture observations have been used to study the process of tight junction formation under various experimental conditions in monolayers of the canine kidney epithelial cell line MDCK. Cells derived from previously confluent cultures and plated immediately after trypsin- EDTA dissociation develop a resistance that reaches its maximum value of several hundred ohms-cm(2) after approximately 24 h and falls to a steady-state value of 80-150 ohms- cm(2) by 48 h. The rise in resistance and the development of tight junctions can be completely and reversibly prevented by the addition of 10 μg/ml cycloheximide at the time of plating, but not when this inhibitor is added more than 10 h after planting. Thus tight junction formation consists of separable synthetic and assembly phases. These two phases can also be dissociated and the requirement for protein synthesis after plating eliminated if, following trypsinization, the cells are maintained in spinner culture for 24 h before plating. The requirement for protein synthesis is restored, however, if cells maintained in spinner culture are treated with trypsin before plating. Actinomycin D prevents development of resistance only in monolayers formed from cells derived from sparse rather than confluent cultures, but new mRNA synthesis is not required if cells obtained from sparse cultures are maintained for 24 h in spinner culture before plating. Once a steady-state resistance has been reached, its maintenance does not require either mRNA or protein synthesis; in fact, inhibition of protein synthesis causes a rise in the resistance over a 30-h period. Following treatments that disrupt the junctions in steady- state monolayers recovery of resistance also does not require protein synthesis. These observations suggest that proteins are involved in tight junction formation. Such proteins, which do not turn over rapidly under steady-state conditions, are destroyed by trypsinization and can be resynthesized in the absence of stable cell-cell or cell-substratum contact. Messenger RNA coding for proteins involved in tight junction formation is stable except when cells are sparsely plated, and can also be synthesized without intercellular contacts or cell-substratum attachment.

scholarly journals Temperature uniformity in the CERN CLOUD chamber

2017 ◽  
Vol 10 (12) ◽  
pp. 5075-5088 ◽  
Author(s):  
António Dias ◽  
Sebastian Ehrhart ◽  
Alexander Vogel ◽  
Christina Williamson ◽  
João Almeida ◽  
...  
Keyword(s):  
Steady State ◽  
Air Temperature ◽  
Elevated Temperatures ◽  
Trace Gases ◽  
Vertical Direction ◽  
Cloud Chamber ◽  
Temperature Uniformity ◽  
Atmospheric Conditions ◽  
Cloud Droplets ◽  
Experimental Conditions

Abstract. The CLOUD (Cosmics Leaving OUtdoor Droplets) experiment at CERN (European Council for Nuclear Research) investigates the nucleation and growth of aerosol particles under atmospheric conditions and their activation into cloud droplets. A key feature of the CLOUD experiment is precise control of the experimental parameters. Temperature uniformity and stability in the chamber are important since many of the processes under study are sensitive to temperature and also to contaminants that can be released from the stainless steel walls by upward temperature fluctuations. The air enclosed within the 26 m3 CLOUD chamber is equipped with several arrays (strings) of high precision, fast-response thermometers to measure its temperature. Here we present a study of the air temperature uniformity inside the CLOUD chamber under various experimental conditions. Measurements were performed under calibration conditions and run conditions, which are distinguished by the flow rate of fresh air and trace gases entering the chamber at 20 and up to 210 L min−1, respectively. During steady-state calibration runs between −70 and +20 °C, the air temperature uniformity is better than ±0.06 °C in the radial direction and ±0.1 °C in the vertical direction. Larger non-uniformities are present during experimental runs, depending on the temperature control of the make-up air and trace gases (since some trace gases require elevated temperatures until injection into the chamber). The temperature stability is ±0.04 °C over periods of several hours during either calibration or steady-state run conditions. During rapid adiabatic expansions to activate cloud droplets and ice particles, the chamber walls are up to 10 °C warmer than the enclosed air. This results in temperature differences of ±1.5 °C in the vertical direction and ±1 °C in the horizontal direction, while the air returns to its equilibrium temperature with a time constant of about 200 s.

Faster adjustment of O2 uptake to the energy requirement of exercise in the trained state

1978 ◽  
Vol 44 (6) ◽  
pp. 877-881 ◽  
Author(s):  
R. C. Hickson ◽  
H. A. Bomze ◽  
J. O. Hollozy
Keyword(s):  
Steady State ◽  
Exercise Training ◽  
Training Program ◽  
Endurance Exercise ◽  
Time Course ◽  
Energy Requirement ◽  
Constant Load ◽  
O2 Uptake ◽  
Endurance Exercise Training

The purpose of this study was to determine the effect of endurance exercise training on the time course of the increase in VO2 toward steady state in response to submaximal constant load work. Seven men participated in a strenuous program of endurance exercise for 40 min/day, 6 days/wk for 10 wk. Their average VO2max increased from 3.29 liters before training to 4.53 liters at the end of the training program. VO2 was measured continuously on a breath-by-breath basis at work rates requiring 40%, 50%, 60%, or 70% of VO2max before training. After training the subjects were retested both at the same absolute and the same relative work rates. The increases in VO2 toward steady state occurred more rapidly in the trained than in the untrained state both at the same absolute and at the same relative work rates. The finding that O2 uptake rises to meet O2 demand more rapidly in the trained than in the untrained state provides evidence that the working muscles become less hypoxic at the onset of exercise of the same intensity after training.

scholarly journals Role of Mammalian Auditory Cortex in the Perception of Elementary Sound Properties

2001 ◽  
Vol 85 (6) ◽  
pp. 2350-2358 ◽  
Author(s):  
Sanjiv K. Talwar ◽  
Pawel G. Musial ◽  
George L. Gerstein
Keyword(s):  
Auditory Cortex ◽  
Time Course ◽  
Auditory Evoked Potentials ◽  
Mammalian Species ◽  
Sound Intensity ◽  
Primary Auditory Cortex ◽  
Normal Hearing ◽  
Auditory Task ◽  
Experimental Conditions

Studies in several mammalian species have demonstrated that bilateral ablations of the auditory cortex have little effect on simple sound intensity and frequency-based behaviors. In the rat, for example, early experiments have shown that auditory ablations result in virtually no effect on the rat's ability to either detect tones or discriminate frequencies. Such lesion experiments, however, typically examine an animal's performance some time after recovery from ablation surgery. As such, they demonstrate that the cortex is not essential for simple auditory behaviors in the long run. Our study further explores the role of cortex in basic auditory perception by examining whether the cortex is normally involved in these behaviors. In these experiments we reversibly inactivated the rat primary auditory cortex (AI) using the GABA agonist muscimol, while the animals performed a simple auditory task. At the same time we monitored the rat's auditory activity by recording auditory evoked potentials (AEP) from the cortical surface. In contrast to lesion studies, the rapid time course of these experimental conditions preclude reorganization of the auditory system that might otherwise compensate for the loss of cortical processing. Soon after bilateral muscimol application to their AI region, our rats exhibited an acute and profound inability to detect tones. After a few hours this state was followed by a gradual recovery of normal hearing, first of tone detection and, much later, of the ability to discriminate frequencies. Surface muscimol application, at the same time, drastically altered the normal rat AEP. Some of the normal AEP components vanished nearly instantaneously to unveil an underlying waveform, whose size was related to the severity of accompanying behavioral deficits. These results strongly suggest that the cortex is directly involved in basic acoustic processing. Along with observations from accompanying multiunit experiments that related the AEP to AI neuronal activity, our results suggest that a critical amount of activity in the auditory cortex is necessary for normal hearing. It is likely that the involvement of the cortex in simple auditory perceptions has hitherto not been clearly understood because of underlying recovery processes that, in the long-term, safeguard fundamental auditory abilities after cortical injury.

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