Creatine Kinase-1 is Principally Inactivated in Serum by Complexing with Immunoglobulin-G

1980 ◽  
Vol 58 (2) ◽  
pp. 157-160 ◽  
Author(s):  
D. A. Nealon ◽  
A. R. Henderson
Keyword(s):  
Creatine Kinase ◽  
Enzyme Activity ◽  
Column Chromatography ◽  
Immunoglobulin G ◽  
Kinase Inhibitors ◽  
Published Data ◽  
Other Substances ◽  
Total Creatine ◽  
The Difference ◽  
The Stability

1. The stability of enzyme activity of the creatine kinase-l-immunoglobulin-G complex has been determined at 37°C, 4°C and −20°C in heat-inactivated serum and buffer. 2. The complex was formed by incubating creatine kinase-1 and immunoglobulin G at 37°C for 30 min. It was isolated by Sephadex G-100 column chromatography. 3. At all temperatures the complex suspended in buffer was about twice as stable as it was in heat-inactivated serum. Stability decreased in the sequence of 4°C, 37°C and −20°C. Therefore all isolations of the complex were carried out at 4°C. 4. At 37°C the decay of enzyme activity of the complex was found to be biphasic first order. In serum the Kd values were −0·045 min−1 and −0·0028 min−1, and in buffer −0·023 min−1 and −0·0016 min−1. 5. From column-chromatography experiments it was found that between 10 and 20% of the total creatine kinase-1 was involved in the complexing reaction after a 30-min incubation. 6. With this 10·20% proportion it can be calculated that the overall t0·5 for the decay of total creatine kinase-1 activity must be in the range 95–201 min. This finding suggests, by comparison with other published data, that the enzyme—immunoglobulin complex is the main route of creatine kinase-1 catabolism in serum. 7. The difference between serum and buffer decay values for the complex is possibly due to the presence of cystine, urate and other substances in serum, which are additional potent creatine kinase inhibitors.

Metastable creatine kinase MM and complexes of creatine kinase BB and immunoglobulin G, "atypical" isoenzymes with similar electrophoretic mobility.

1980 ◽  
Vol 26 (1) ◽  
pp. 78-83
Author(s):  
H A Homburger ◽  
L Wold ◽  
G L Jacob ◽  
J O'Brien
Keyword(s):  
Creatine Kinase ◽  
Electrophoretic Mobility ◽  
Immunoglobulin G ◽  
Gradient Elution ◽  
Creatine Kinase Activity ◽  
B Subunit ◽  
Creatine Kinase B ◽  
Agarose Electrophoresis ◽  
Total Creatine Kinase Activity ◽  
Total Creatine

Abstract Metastable creatine kinase MM isoenzyme was isolated and partially purified from homogenates of myocardium and skeletal muscle by gradient elution on carboxymethyl cellulose. This variant isoenzyme migrated between the MM and MB isoenzymes on agarose electrophoresis, accounted for 3.5% of the total creatine kinase activity in each tissue, was not a macromolecule, and had stable electrophoretic mobility only in borate buffer (0.02 mol/L). By comparison, the creatine kinase isoenzymes with similar "atypical" electrophoretic mobility in serum specimens were complexes of the BB isoenzyme and immunoglobulin G. These complexes were measured by a radioimmunoassay specific for the creatine kinase B-subunit and eluted predominantly with the MB isoenzyme in a commercial anion-exchange reagent system.

Analysis of metabolic control: new insights using scaled creatine kinase model

1988 ◽  
Vol 254 (6) ◽  
pp. R949-R959 ◽  
Author(s):  
R. J. Connett
Keyword(s):  
Creatine Kinase ◽  
Adenine Nucleotide ◽  
Adenine Nucleotides ◽  
Binding Constants ◽  
Cation Binding ◽  
Ph Values ◽  
Total Phosphate ◽  
Total Creatine ◽  
The Stability ◽  
Adenine Nucleotide Pool

The creatine kinase-adenylate kinase equilibria equations are given a dimensionless form by normalizing to total creatine concentration. Analysis with appropriate equilibrium and cation-binding constants identified two sharply separated phases of energy depletion. In the "buffering" phase, energy is derived from phosphocreatine. In the "depleting" phase, adenine nucleotides are the source of energy. Defining the state of the adenine nucleotide pool requires only pH, phosphocreatine, and creatine concentrations. Analysis of data from skeletal muscle, heart, brain, and smooth muscle demonstrated that the [free adenine nucleotide]/[total creatine] and [total phosphate]/[total creatine] are essentially constant over the greater than 20-fold concentration range among tissues and species. This result permits quantitative evaluation of cell energetics with data scaled to the total phosphate, as obtained with nuclear magnetic resonance studies, or to total creatine, as obtained in chemical analysis of freeze-trapped tissue. By applying the stability of the tissue parameters to the equations, it is demonstrated that unique identification of a hypothesis describing the recruitment of O2 uptake requires testing at several pH values.

scholarly journals Different sensitivity of DNA in situ in interphase and metaphase chromatin to heat denaturation.

1977 ◽  
Vol 73 (1) ◽  
pp. 128-138 ◽  
Author(s):  
Z Darzynkiewicz ◽  
F Traganos ◽  
T Sharpless ◽  
M R Melamed
Keyword(s):  
Heat Denaturation ◽  
Published Data ◽  
Dna Denaturation ◽  
Metaphase Chromosomes ◽  
Flow Cytofluorometry ◽  
Dna Reassociation ◽  
Interphase Cells ◽  
The Difference ◽  
The Stability

Heat denaturation of DNA in situ, in unbroken cells, was studied in relation to the cell cycle. DNA in metaphase cells denatured at lower temperatures (8 degrees-10 degrees C lower) than DNA in interphase cells. Among interphase cells, small differences between G1, S, and G2 cells were observed at temperatures above 90 degrees C. The difference between metaphase and interphase cells increased after short pretreatment with formaldehyde, decreased when cells were heated in the presence of 1 mM MgCl2, and was abolished by cell pretreatment with 0.5 N HCl. The results suggest that acid-soluble constituents of chromatin confer local stability to DNA and that the degree of stabilization is lower in metaphase chromosomes than in interphase nuclei. These in situ results remain in contrast to the published data showing no difference in DNA denaturation in chromatin isolated from interphase and metaphase cells. It is likely that factors exist which influence the stability of DNA in situ are associated with the super-structural organization of chromatin in intact nuclei and which are lost during chromatin isolation and solubilization. Since DNA denaturation is assayed after cell cooling, there is also a possibility that the extent of denatured DNA may be influenced by some factors that control strand separation and DNA reassociation. The different stainability of interphase vs. metaphase cells, based on the difference in stability of DNA, offers a method for determining mitotic indices by flow cytofluorometry, and a possible new parameter for sorting cells in metaphase.

Stability of commonly used thiols and of human creatine kinase isoenzymes during storage at various temperature in various media.

1977 ◽  
Vol 23 (5) ◽  
pp. 816-829 ◽  
Author(s):  
D A Nealon ◽  
A R Henderson
Keyword(s):  
Creatine Kinase ◽  
Fluorescent Light ◽  
Human Origin ◽  
Creatine Kinase Isoenzyme ◽  
Aqueous Buffer ◽  
Creatine Kinase Isoenzymes ◽  
Total Creatine ◽  
The Stability ◽  
Sulfhydryl Compounds ◽  
Fast Freezing

Abstract We assessed the stability--at 20, 4 --20, and --80 degrees C--of 2-mercaptoethanol, dithioerythritol, and dithiothreitol, and of semi-purified creatine kinase isoenzymes of human origin in the presence and absence of the three compounds. The isoenzymes plus the sulfhydryl compounds were also assessed an aqueous buffer, in heat-inactivated pooled sera, or in fresh pooled sera with low endogenous total creatine kinase. We also examined the influence of thawing conditions from--20 or--80 degrees C, of fast freezing, and of exposure to fluorescent light. At 20 and 4 degrees C some--SH groups are oxidized. At--20 and--80 degrees C this loss is diminished. It is a function of both temperature and diluent. For stability, creative kinase isoenzymes stored at any temperature require the presence of a suitable sulfhydryl compound. Periodic addition of fresh sulfhydryl solutions stabilized creatine kinase isoenzymes suspended in the two protein-based diluents by 10-30%; for those in an aqueous buffer such additions decreased the activity to stored creatine kinase isoenzymes. Fast freezing does not increase recovery of creatine kinase isoenzyme activity over slow freezing at--80 or--20 degrees C. Thawing should be done as infrequently as possible and should be done rapidly, at 37 degrees C, for the residual enzyme activity to be maximal.

Metastable creatine kinase MM and complexes of creatine kinase BB and immunoglobulin G, "atypical" isoenzymes with similar electrophoretic mobility.

1980 ◽  
Vol 26 (1) ◽  
pp. 78-83 ◽  
Author(s):  
H A Homburger ◽  
L Wold ◽  
G L Jacob ◽  
J O'Brien
Keyword(s):  
Creatine Kinase ◽  
Electrophoretic Mobility ◽  
Immunoglobulin G ◽  
Gradient Elution ◽  
Creatine Kinase Activity ◽  
B Subunit ◽  
Creatine Kinase B ◽  
Agarose Electrophoresis ◽  
Total Creatine Kinase Activity ◽  
Total Creatine

Abstract Metastable creatine kinase MM isoenzyme was isolated and partially purified from homogenates of myocardium and skeletal muscle by gradient elution on carboxymethyl cellulose. This variant isoenzyme migrated between the MM and MB isoenzymes on agarose electrophoresis, accounted for 3.5% of the total creatine kinase activity in each tissue, was not a macromolecule, and had stable electrophoretic mobility only in borate buffer (0.02 mol/L). By comparison, the creatine kinase isoenzymes with similar "atypical" electrophoretic mobility in serum specimens were complexes of the BB isoenzyme and immunoglobulin G. These complexes were measured by a radioimmunoassay specific for the creatine kinase B-subunit and eluted predominantly with the MB isoenzyme in a commercial anion-exchange reagent system.

Use of a Lyophilized Reference Plasma to Compare Coagulation Test Procedures

1975 ◽  
Vol 34 (02) ◽  
pp. 426-444 ◽  
Author(s):  
J Kahan ◽  
I Nohén
Keyword(s):  
Oral Anticoagulant Therapy ◽  
Repeated Measurements ◽  
Test Procedures ◽  
Coagulation Activity ◽  
Close Relationship ◽  
Measured Activity ◽  
Test Materials ◽  
The Difference ◽  
The Stability ◽  
The One

SummaryIn 4 collaborative trials, involving a varying number of hospital laboratories in the Stockholm area, the coagulation activity of different test materials was estimated with the one-stage prothrombin tests routinely used in the laboratories, viz. Normotest, Simplastin-A and Thrombotest. The test materials included different batches of a lyophilized reference plasma, deep-frozen specimens of diluted and undiluted normal plasmas, and fresh and deep-frozen specimens from patients on long-term oral anticoagulant therapy.Although a close relationship was found between different methods, Simplastin-A gave consistently lower values than Normotest, the difference being proportional to the estimated activity. The discrepancy was of about the same magnitude on all the test materials, and was probably due to a divergence between the manufacturers’ procedures used to set “normal percentage activity”, as well as to a varying ratio of measured activity to plasma concentration. The extent of discrepancy may vary with the batch-to-batch variation of thromboplastin reagents.The close agreement between results obtained on different test materials suggests that the investigated reference plasma could be used to calibrate the examined thromboplastin reagents, and to compare the degree of hypocoagulability estimated by the examined PIVKA-insensitive thromboplastin reagents.The assigned coagulation activity of different batches of the reference plasma agreed closely with experimentally obtained values. The stability of supplied batches was satisfactory as judged from the reproducibility of repeated measurements. The variability of test procedures was approximately the same on different test materials.

In the stability of paints and varnishes for protective and decorative wood finish

2016 ◽  
Author(s):  
Ю.И. Цой
Keyword(s):  
Organic Solvents ◽  
Evaluation Criteria ◽  
Ternary Systems ◽  
Solubility Parameters ◽  
Formaldehyde Resin ◽  
Film Forming ◽  
The Difference ◽  
Molecular Forces ◽  
The Stability ◽  
Thermodynamic Interaction

Задача получения однородного состава ЛКМ сводится к правильному выбору пленкообразователей и соответствующих растворителей и, в случае необходимости, разбавителей и добавок. На основе положений теории растворимости была проведена оценка совместимости компонентов и стабильности таких систем. Критериями оценки были приняты параметры растворимости и термодинамического взаимодействия. Исследовали совместимость алкидных лаков с аминоформальдегидными смолами в бутаноле, и результаты этих исследований представлены в треугольной фазовой диаграмме. Для оценки совместимости смол по результатам расчета параметров термодинамического взаимодействия были построены спинодали для тройных систем. Как показывает анализ результатов проведенных исследований, даже при большой разности ?? = |?12 – ?13| = 0,2 наблюдается небольшая область несовместимости. С уменьшением разности ?? область несовместимости сужается. При ?23 > ?с область несовместимости расширяется как для ? = 0,2, так и для ?? = 0,1 и ?? = 0. Таким образом, для таких систем на совместимость компонентов, в первую очередь, влияет их взаимодействие между собой – чем оно больше, тем лучше они совместимы. Проведенные исследования показали, что парциальные параметры растворимости, учитывающие природу когезионных сил, более объективно характеризуют растворимость пленкообразующих смол в органических растворителях, чем их общий параметр растворимости. Растворимость пленкообразующих алкидных и этерифицированных аминоформальдегидных смол в органических растворителях обусловлена термодинамическим взаимодействием молекулярных сил различной природы. Стабильность смеси алкидной и этерифицированной аминоформальдегидной смолы в бутаноле обусловлена в большей степени термодинамическим взаимодействием молекулярных сил; при этом степень бутанолизации меламиноформальдегидной смолы оказывает наибольшее влияние на стабильность cистемы. Проведенные исследования по стабильности лакокрасочных смесей из различных пленкообразующих смол на основе рассмотренных критериев оценки могут быть использованы для совершенствования технологии защитно-декоративной отделки древесины. The problem of obtaining a homogeneous composition of the coatings is reduced to the proper selection of film-forming agents, and diluent and, if appropriate, diluents and additives. On the basis of the theory of solubility, we have carried out the assessment of the compatibility of the components and stability of such systems. The evaluation criteria were adopted the parameters of the solubility and thermodynamic interaction. Researched the compatibility with alkyd varnishes aminoformaldehyde resin in butanol, and the results of these studies are presented in the triangular phase diagram. To evaluate the compatibility of the resins according to the results of calculation of thermodynamic parameters of interaction were built spinodal for ternary systems. As the analysis of the results of the studies, even when the difference ??|?12 – ?13| = 0,2 there is a small area of incompatibility. With the decrease of the difference ? ? region incompatibility narrows. When ?23> ?с region incompatibility extends to ?? = 0,2, and for ?? = 0.1 and ?? = 0. Thus, for such systems, compatibility of components, primarily, affects their interaction with each other – the further away it is, the better they are compatible. Studies have shown that the partial solubility parameters, which take into account the nature of cohesive forces, more objectively characterize the solubility of film-forming resins in organic solvents than their total solubility parameter. The solubility and film-forming aminoformaldehyde esterified alkyd resins in organic solvents due to thermodynamic interaction of molecular forces of different nature. Тhe stability of the mixture and aminoformaldehyde esterified alkyd resin in butanol is generated largely by thermodynamic forces of molecular interaction; the degree of butanolate melamine-formaldehyde resin has the greatest effect on the stability of the system. Conducted research on the stability of the paint mixtures of various film-forming resins on the basis of the evaluation criteria can be used to improve the technology of protective and decorative wood finish.

Stability and mutual impact of perception of parental pressure and infants' social support – A study based on the difference between employment status and household income

2021 ◽  
pp. 002072092199705
Author(s):  
Yong Wang
Keyword(s):  
Household Income ◽  
Employment Status ◽  
Working Mothers ◽  
Social Research ◽  
Research Report ◽  
Parental Pressure ◽  
Income Levels ◽  
Significant Difference ◽  
The Difference ◽  
The Stability

The purpose of this study is to explore the stability and interaction between parental pressure and social research report, as well as the role of employment status and family income levels in this process. This study used a special study on Korean children (PSKC) 2–4 waves. Use t-test, correlation and autoregressive cross-delay modeling to analyze the data. The main findings of this study are: First, over time, parental pressure and mother’s social research report are consistent. Secondly, the pressure of motherhood and childcare has an obvious lagging effect on upbringing, and vice versa. Third, there is no significant difference between working mothers and non-working mothers in terms of the stability of working parents' pressure, social research report and social research report for children's pressure channels. However, parental pressure can only predict the social research report of working mothers. Fourth, there is no significant difference between the stability and interaction of these two structures in household income levels. In short, the results show that, over time, parental pressure is consistent with mother’s social research report. The results also show that there is a significant cross-lag effect between the mothers’ perceptions of mutual pressure analysis. In the process from parental pressure to social research report, I found the difference between working and non-working mothers. The advantage of this study is that the expected longitudinal design was adopted during infancy and the priority between the two structures can be considered. The results of this study can be used as a source of intervention plans to help parents withstand severe parenting pressure and lack of social research report.

scholarly journals Modeling and Analysis of BDS-2 and BDS-3 Combined Precise Time and Frequency Transfer Considering Stochastic Models of Inter-System Bias

2021 ◽  
Vol 13 (4) ◽  
pp. 793
Author(s):  
Guoqiang Jiao ◽  
Shuli Song ◽  
Qinming Chen ◽  
Chao Huang ◽  
Ke Su ◽  
...  
Keyword(s):  
Stochastic Models ◽  
Satellite System ◽  
Modeling And Analysis ◽  
System Bias ◽  
Frequency Transfer ◽  
Global Navigation ◽  
The Difference ◽  
The Stability ◽  
Global Navigation Satellite ◽  
Precise Time

BeiDou global navigation satellite system (BDS) began to provide positioning, navigation, and timing (PNT) services to global users officially on 31 July, 2020. BDS constellations consist of regional (BDS-2) and global navigation satellites (BDS-3). Due to the difference of modulations and characteristics for the BDS-2 and BDS-3 default civil service signals (B1I/B3I) and the increase of new signals (B1C/B2a) for BDS-3, a systemically bias exists in the receiver-end when receiving and processing BDS-2 and BDS-3 signals, which leads to the inter-system bias (ISB) between BDS-2 and BDS-3 on the receiver side. To fully utilize BDS, the BDS-2 and BDS-3 combined precise time and frequency transfer are investigated considering the effect of the ISB. Four kinds of ISB stochastic models are presented, which are ignoring ISB (ISBNO), estimating ISB as random constant (ISBCV), random walk process (ISBRW), and white noise process (ISBWN). The results demonstrate that the datum of receiver clock offsets can be unified and the ISB deduced datum confusion can be avoided by estimating the ISB. The ISBCV and ISBRW models are superior to ISBWN. For the BDS-2 and BDS-3 combined precise time and frequency transfer using ISBNO, ISBCV, ISBRW, and ISBWN, the stability of clock differences of old signals can be enhanced by 20.18%, 23.89%, 23.96%, and 11.46% over BDS-2-only, respectively. For new signals, the enhancements are −50.77%, 20.22%, 17.53%, and −3.69%, respectively. Moreover, ISBCV and ISBRW models have the better frequency transfer stability. Consequently, we recommended the optimal ISBCV or suboptimal ISBRW model for BDS-2 and BDS-3 combined precise time and frequency transfer when processing the old as well as the new signals.

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