Temperature dependence of oxygen diffusion and consumption in mammalian striated muscle

1993 ◽  
Vol 264 (6) ◽  
pp. H1825-H1830 ◽  
Author(s):  
T. B. Bentley ◽  
H. Meng ◽  
R. N. Pittman
Keyword(s):  
Diffusion Coefficient ◽  
Steady State ◽  
Oxygen Diffusion ◽  
Striated Muscle ◽  
Effect Of Temperature ◽  
Retractor Muscle ◽  
Oxygen Solubility ◽  
Order Of Magnitude

This study investigated the effect of temperature on the oxygen diffusion coefficient (DO2) of hamster retractor muscle from 11 to 37 degrees C. DO2 was measured using a non-steady-state technique, whereas muscle O2 consumption (VO2) was estimated after steady state was reached. DO2 was 0.84 +/- 0.04 x 10(-5) cm2/s at 11 degrees C and rose exponentially to 2.41 +/- 0.19 x 10(-5) cm2/s at 37 degrees C, producing a temperature coefficient for DO2 of 4.60%/degrees C for this temperature range. To measure DO2 directly at 37 degrees C, it was necessary to inhibit tissue VO2 with Amytal. The DO2 measurements made at 37 degrees C were significantly higher than previously reported values, which had been based on extrapolations from lower temperatures (6). Further analysis suggests a possible transition in the diffusion pathway between 23 and 30 degrees C, resulting in a DO2 higher than that previously expected. This larger DO2, together with a recently published value of oxygen solubility (alpha) (21), results in an in vitro Krogh's diffusion coefficient (KO2) that is 2.4 times larger than that previously reported (24) and therefore significantly reduces an order of magnitude discrepancy between in vitro and estimated in vivo KO2 values (24). Muscle VO2 was 0.35 ml O2.min-1.100 g-1 at 11 degrees C and increased with temperature, resulting in an activation energy of the rate-limiting reaction from the Arrhenius equation of -10.5 kcal/mol between 11 and 30 degrees C.

Oxygen diffusion in hamster striated muscle: comparison of in vitro and near in vivo conditions

1992 ◽  
Vol 263 (1) ◽  
pp. H35-H39 ◽  
Author(s):  
H. Meng ◽  
T. B. Bentley ◽  
R. N. Pittman
Keyword(s):  
Diffusion Coefficient ◽  
Striated Muscle ◽  
Statistical Significance ◽  
Retractor Muscle ◽  
Group I ◽  
Arterial Inflow ◽  
Order Of Magnitude ◽  
O2 Diffusion

To investigate the suggestion of A. S. Popel, R. N. Pittman, and M. L. Ellsworth [Am. J. Physiol. 256 (Heart Circ. Physiol. 25): H921-H924, 1989] that Krogh's diffusion coefficient for O2 in vivo might be an order of magnitude higher than in vitro, O2 diffusion coefficient (DO2) and resting O2 consumption were measured on hamster retractor muscle in vitro and under near in vivo conditions where the muscle remained attached to the animal but the arterial inflow was occluded just before measurement. Experiments were performed on two groups of animals, differing in weight and age. We found that DO2 determined in vitro (extrapolated to 37 degrees C) was 1.81 +/- 0.12 x 10(-5) cm2/s for group I (smaller and younger), which was not significantly different from the value (2.00 +/- 0.08 x 10(-5) cm2/s) determined in group II. In both groups, DO2 under near in vivo conditions tended to be 10-15% larger than the value in vitro, although this trend did not reach statistical significance. It is unlikely that this trend is large enough to reconcile the inconsistency between theoretical and experimental determinations of O2 diffusion from the arteriolar network of this tissue.

Influence of temperature on oxygen diffusion in hamster retractor muscle

1997 ◽  
Vol 272 (3) ◽  
pp. H1106-H1112 ◽  
Author(s):  
T. B. Bentley ◽  
R. N. Pittman
Keyword(s):  
Diffusion Coefficient ◽  
Lipid Membranes ◽  
Tissue Oxygenation ◽  
Retractor Muscle ◽  
Hyperbaric Chamber ◽  
Influence Of Temperature On ◽  
Order Of Magnitude ◽  
O2 Diffusion

A mathematical analysis by Popel et al. [Am. J. Physiol. 256 (Heart Circ. Physiol. 25): H921-H924, 1989] of in vivo data on arteriolar O2 loss suggested that Krogh's diffusion coefficient (KO2 = alpha x DO2, where DO2 is the O2 diffusion coefficient and alpha is the tissue O2 solubility) in vivo could be an order of magnitude larger than that calculated from DO2 values measured in vitro at 22 degrees C and extrapolated to 37 degrees C. In this study, to eliminate potential extrapolation errors, we used a miniature hyperbaric chamber with 1-2 atm of O2 to maintain tissue oxygenation and allow DO2 measurements directly at 37 degrees C while using a non-steady-state technique. The need for metabolic poisons that had been required by earlier experimental techniques was thereby eliminated. DO2 measured directly at 37 degrees C (2.42 x 10(-5) cm2/s) and the increase with temperature of DO2 between 30 and 41 degrees C (4.61%/degrees C) were unexpectedly higher than the values we found at lower temperatures. Oxygen consumption was also higher than expected at 37 degrees and 40 degrees C. An analysis of the activation energy for DO2 suggests that at higher temperatures there is a change in the diffusion pathway from that existing at lower temperatures, perhaps caused by phase transitions in the lipid membranes.

scholarly journals Three Kinetically Distinct Ca2+-Independent Depolarization-Activated K+ Currents in Callosal-Projecting Rat Visual Cortical Neurons

1997 ◽  
Vol 78 (5) ◽  
pp. 2309-2320 ◽  
Author(s):  
Rachel E. Locke ◽  
Jeanne M. Nerbonne
Keyword(s):  
Steady State ◽  
Cortical Neurons ◽  
Differential Sensitivity ◽  
Kinetic Properties ◽  
Time Constants ◽  
Steady State Inactivation ◽  
Order Of Magnitude ◽  
Visual Cortical

Locke, Rachel E. and Jeanne M. Nerbonne. Three kinetically distinct Ca2+-independent depolarization-activated K+ currents in callosal-projecting rat visual cortical neurons. J. Neurophysiol. 78: 2309–2320, 1997. Whole cell, Ca2+-independent, depolarization-activated K+ currents were characterized in identified callosal-projecting (CP) neurons isolated from postnatal day 7–16 rat primary visual cortex. CP neurons were identified in vitro after in vivo retrograde labeling with fluorescently tagged latex microbeads. During brief (160-ms) depolarizing voltage steps to potentials between −50 and +60 mV, outward K+ currents in these cells activate rapidly and inactivate to varying degrees. Three distinct K+ currents were separated based on differential sensitivity to 4-aminopyridine (4-AP); these are referred to here as I A, I D, and I K, because their properties are similar (but not identical) K+ currents termed I A, I D, and I K in other cells. The current sensitive to high (≥100 μM) concentrations of 4-AP ( I A) activates and inactivates rapidly; the current blocked completely by low (≤50 μM) 4-AP ( I D) activates rapidly and inactivates slowly. A slowly activating, slowly inactivating current ( I K) remains in the presence of 5 mM 4-AP. I A, I D, and I K also were separated and characterized in experiments that did not rely on the use of 4-AP. All CP cells express all three K+ current types, although the relative densities of I A, I D, and I K vary among cells. The experiments here also have revealed that I A, I D, and I K display similar voltage dependences of activation and steady state inactivation, whereas the kinetic properties of the currents are distinct. At +30 mV, for example, mean ± SD activation τs are 0.83 ± 0.24 ms for I A, 1.74 ± 0.49 ms for I D, and 14.7 ± 4.0 ms for I K. Mean ± SD inactivation τs for I A and I D are 26 ± 7 ms and 569 ± 143 ms, respectively. Inactivation of I K is biexponential with mean ± SD inactivation time constants of 475 ± 232 ms and 3,128 ± 1,328 ms; ∼20% of the 4-AP–insensitive current is noninactivating. For all three components, activation is voltage dependent, increasing with increasing depolarization, whereas inactivation is voltage independent. Both I A and I K recover rapidly from steady state inactivation with mean ± SD recovery time constants of 38 ± 7 ms and 79 ± 26 ms, respectively; I D recovers an order of magnitude more slowly (588 ± 274 ms). The properties of I A, I D, and I K in CP neurons are compared with those of similar currents described previously in other mammalian central neurons and, in the accompanying paper, the roles of these conductances in regulating the firing properties of CP neurons are explored.

Applicability of Instability Index for In vitro Protein Stability Prediction

2019 ◽  
Vol 26 (5) ◽  
pp. 339-347 ◽  
Author(s):  
Dilani G. Gamage ◽  
Ajith Gunaratne ◽  
Gopal R. Periyannan ◽  
Timothy G. Russell
Keyword(s):  
Protein Stability ◽  
Caulobacter Crescentus ◽  
Effect Of Temperature ◽  
Instability Index ◽  
Dipeptide Composition ◽  
Experimental Conditions ◽  
The Stability ◽  
Vitro Protein

Background: The dipeptide composition-based Instability Index (II) is one of the protein primary structure-dependent methods available for in vivo protein stability predictions. As per this method, proteins with II value below 40 are stable proteins. Intracellular protein stability principles guided the original development of the II method. However, the use of the II method for in vitro protein stability predictions raises questions about the validity of applying the II method under experimental conditions that are different from the in vivo setting. Objective: The aim of this study is to experimentally test the validity of the use of II as an in vitro protein stability predictor. Methods: A representative protein CCM (CCM - Caulobacter crescentus metalloprotein) that rapidly degrades under in vitro conditions was used to probe the dipeptide sequence-dependent degradation properties of CCM by generating CCM mutants to represent stable and unstable II values. A comparative degradation analysis was carried out under in vitro conditions using wildtype CCM, CCM mutants and two other candidate proteins: metallo-β-lactamase L1 and α -S1- casein representing stable, borderline stable/unstable, and unstable proteins as per the II predictions. The effect of temperature and a protein stabilizing agent on CCM degradation was also tested. Results: Data support the dipeptide composition-dependent protein stability/instability in wt-CCM and mutants as predicted by the II method under in vitro conditions. However, the II failed to accurately represent the stability of other tested proteins. Data indicate the influence of protein environmental factors on the autoproteolysis of proteins. Conclusion: Broader application of the II method for the prediction of protein stability under in vitro conditions is questionable as the stability of the protein may be dependent not only on the intrinsic nature of the protein but also on the conditions of the protein milieu.

scholarly journals Ultra-strong bio-glue from genetically engineered polypeptides

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chao Ma ◽  
Jing Sun ◽  
Bo Li ◽  
Yang Feng ◽  
Yao Sun ◽  
...  
Keyword(s):  
Soft Tissues ◽  
Covalent Bond ◽  
Genetically Engineered ◽  
Supramolecular Interactions ◽  
Molar Ratio ◽  
High Adhesion ◽  
Strong Adhesion ◽  
Order Of Magnitude

AbstractThe development of biomedical glues is an important, yet challenging task as seemingly mutually exclusive properties need to be combined in one material, i.e. strong adhesion and adaption to remodeling processes in healing tissue. Here, we report a biocompatible and biodegradable protein-based adhesive with high adhesion strengths. The maximum strength reaches 16.5 ± 2.2 MPa on hard substrates, which is comparable to that of commercial cyanoacrylate superglue and higher than other protein-based adhesives by at least one order of magnitude. Moreover, the strong adhesion on soft tissues qualifies the adhesive as biomedical glue outperforming some commercial products. Robust mechanical properties are realized without covalent bond formation during the adhesion process. A complex consisting of cationic supercharged polypeptides and anionic aromatic surfactants with lysine to surfactant molar ratio of 1:0.9 is driven by multiple supramolecular interactions enabling such strong adhesion. We demonstrate the glue’s robust performance in vitro and in vivo for cosmetic and hemostasis applications and accelerated wound healing by comparison to surgical wound closures.

scholarly journals Phosphorylation-dependent interaction of the synaptic vesicle proteins cysteine string protein and synaptotagmin I

2002 ◽  
Vol 364 (2) ◽  
pp. 343-347 ◽  
Author(s):  
Gareth J.O. EVANS ◽  
Alan MORGAN
Keyword(s):  
Rat Brain ◽  
Direct Interaction ◽  
Secretory Vesicle ◽  
Brain Membrane ◽  
Synaptotagmin I ◽  
Phosphorylated Form ◽  
Order Of Magnitude ◽  
And Function

The secretory vesicle cysteine string proteins (CSPs) are members of the DnaJ family of chaperones, and function at late stages of Ca2+-regulated exocytosis by an unknown mechanism. To determine novel binding partners of CSPs, we employed a pull-down strategy from purified rat brain membrane or cytosolic proteins using recombinant hexahistidine-tagged (His6-)CSP. Western blotting of the CSP-binding proteins identified synaptotagmin I to be a putative binding partner. Furthermore, pull-down assays using cAMP-dependent protein kinase (PKA)-phosphorylated CSP recovered significantly less synaptotagmin. Complexes containing CSP and synaptotagmin were immunoprecipitated from rat brain membranes, further suggesting that these proteins interact in vivo. Binding assays in vitro using recombinant proteins confirmed a direct interaction between the two proteins and demonstrated that the PKA-phosphorylated form of CSP binds synaptotagmin with approximately an order of magnitude lower affinity than the non-phosphorylated form. Genetic studies have implicated each of these proteins in the Ca2+-dependency of exocytosis and, since CSP does not bind Ca2+, this novel interaction might explain the Ca2+-dependent actions of CSP.

scholarly journals Activating Signal Cointegrator 2 Belongs to a Novel Steady-State Complex That Contains a Subset of Trithorax Group Proteins

2003 ◽  
Vol 23 (1) ◽  
pp. 140-149 ◽  
Author(s):  
Young-Hwa Goo ◽  
Young Chang Sohn ◽  
Dae-Hwan Kim ◽  
Seung-Whan Kim ◽  
Min-Jung Kang ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Steady State ◽  
Nuclear Receptors ◽  
Transcriptional Activation ◽  
Retinoic Acid Receptor ◽  
Dependent Manner ◽  
Trithorax Group ◽  
Trithorax Group Proteins

ABSTRACT Many transcription coactivators interact with nuclear receptors in a ligand- and C-terminal transactivation function (AF2)-dependent manner. These include activating signal cointegrator 2 (ASC-2), a recently isolated transcriptional coactivator molecule, which is amplified in human cancers and stimulates transactivation by nuclear receptors and numerous other transcription factors. In this report, we show that ASC-2 belongs to a steady-state complex of approximately 2 MDa (ASC-2 complex [ASCOM]) in HeLa nuclei. ASCOM contains retinoblastoma-binding protein RBQ-3, α/β-tubulins, and trithorax group proteins ALR-1, ALR-2, HALR, and ASH2. In particular, ALR-1/2 and HALR contain a highly conserved 130- to 140-amino-acid motif termed the SET domain, which was recently implicated in histone H3 lysine-specific methylation activities. Indeed, recombinant ALR-1, HALR, and immunopurified ASCOM exhibit very weak but specific H3-lysine 4 methylation activities in vitro, and transactivation by retinoic acid receptor appears to involve ligand-dependent recruitment of ASCOM and subsequent transient H3-lysine 4 methylation of the promoter region in vivo. Thus, ASCOM may represent a distinct coactivator complex of nuclear receptors. Further characterization of ASCOM will lead to a better understanding of how nuclear receptors and other transcription factors mediate transcriptional activation.

scholarly journals TRIM72 is required for effective repair of alveolar epithelial cell wounding

2014 ◽  
Vol 307 (6) ◽  
pp. L449-L459 ◽  
Author(s):  
Seong Chul Kim ◽  
Thomas Kellett ◽  
Shaohua Wang ◽  
Miyuki Nishi ◽  
Nagaraja Nagre ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Molecular Mechanisms ◽  
Striated Muscle ◽  
Alveolar Epithelial Cell ◽  
Alveolar Epithelial Cells ◽  
Lung Cells ◽  
Alveolar Epithelial ◽  
High Tidal Volume Ventilation

The molecular mechanisms for lung cell repair are largely unknown. Previous studies identified tripartite motif protein 72 (TRIM72) from striated muscle and linked its function to tissue repair. In this study, we characterized TRIM72 expression in lung tissues and investigated the role of TRIM72 in repair of alveolar epithelial cells. In vivo injury of lung cells was introduced by high tidal volume ventilation, and repair-defective cells were labeled with postinjury administration of propidium iodide. Primary alveolar epithelial cells were isolated and membrane wounding and repair were labeled separately. Our results show that absence of TRIM72 increases susceptibility to deformation-induced lung injury whereas TRIM72 overexpression is protective. In vitro cell wounding assay revealed that TRIM72 protects alveolar epithelial cells through promoting repair rather than increasing resistance to injury. The repair function of TRIM72 in lung cells is further linked to caveolin 1. These data suggest an essential role for TRIM72 in repair of alveolar epithelial cells under plasma membrane stress failure.

The anterior and posterior ‘stomach’ regions of larval Aedes aegypti midgut: regional specialization of ion transport and stimulation by 5-hydroxytryptamine

1999 ◽  
Vol 202 (3) ◽  
pp. 247-252 ◽  
Author(s):  
T.M. Clark ◽  
A. Koch ◽  
D.F. Moffett
Keyword(s):  
Steady State ◽  
Aedes Aegypti ◽  
Ion Transport ◽  
Malpighian Tubules ◽  
Transport Mechanisms ◽  
Regional Specialization ◽  
Negative Deflection ◽  
Electrical Polarity

The ‘stomach’ region of the larval mosquito midgut is divided into histologically distinct anterior and posterior regions. Anterior stomach perfused symmetrically with saline in vitro had an initial transepithelial potential (TEP) of −66 mV (lumen negative) that decayed within 10–15 min to a steady-state TEP near −10 mV that was maintained for at least 1 h. Lumen-positive TEPs were never observed in the anterior stomach. The initial TEP of the perfused posterior stomach was opposite in polarity, but similar in magnitude, to that of the anterior stomach, measuring +75 mV (lumen positive). This initial TEP of the posterior stomach decayed rapidly at first, then more slowly, eventually reversing the electrical polarity of the epithelium as lumen-negative TEPs were recorded in all preparations within 70 min. Nanomolar concentrations of the biogenic amine 5-hydroxytryptamine (5-HT, serotonin) stimulated both regions, causing a negative deflection of the TEP of the anterior stomach and a positive deflection of the TEP of the posterior stomach. Phorbol 12,13-diacetate also caused a negative deflection of the TEP of the anterior stomach, but had no effect on the TEP of the posterior stomach. These data demonstrate that 5-HT stimulates region-specific ion-transport mechanisms in the stomach of Aedes aegypti and suggest that 5-HT coordinates the actions of the Malpighian tubules and midgut in the maintenance of an appropriate hemolymph composition in vivo.

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