This study was investigated under the postulation that activation of intracellular calcium- calmodulin complex during ischemia-reperfusion leads to myocardial injury. The protective effects of calcium channel blocker, diltiazem and calmodulin inhibitors, trifluoperazine, flunarizine and calmidazolium from ischemic injury in rat hearts were observed by using Langendorff apparatus when the antagonists were infused for 3 min in the beginning of ischemia. Thereby, an increase in resting tension developed during 30-min ischemia was analyzed with regard to [1] the degree of cardiac functional recovery following 60-min reperfusion, [2] changes in biochemical variables evoked during 30-min ischemia. The results obtained were as follows: l. In the ischemic group, the resting tension was increased by 4.1*0.2 g at 30-min ischemia. However, the increase in resting tension was markedly reduced not only by pretreatment with diltiazem [3.3 p M] but also with calmodulin inhibitors, trifluoperazine [3.3 p M], flunarizine [0.5 p M] and calmidazolium [0.5 p M], respectively. 2. Recovery of myocardial contractility, +dF /dt and coronary flow were much reduced when evoked by reperfusion in the ischemic group. These variables were significantly improved either by pretreatment with diltiazem or with calmodulin inhibitors. 3. The resting tension increment evoked during ischemia was significantly inversely correlated with the degree of cardiac function recovered during reperfusion. 4. Following 30-min ischemia, the production of malondialdehyde and release of lysosomal enzyme were much increased in association with a decrease in creatine kinase activity. 5. The increases in malondialdehyde production and release of free lysosomal enzyme were suppressed by pretreatment with calmodulin inhibitors as well as diltiazem. Likewise, the decrease of creatine kinase activities was prevented by these calcium antagonists. With these results, it is indicated that a increase in resting tension observed during ischemia has an inverse relationship to the cardiac function recovered following reperfusion, and further, the later may be significantly dependent on the degree of biochemical alterations occurred during ischemia such as decrease in creatine kinase activity, increased production of malondialdehyde and increased release of free lysosomal enzyme. Thus it is concluded that calmodulin plays a pivotal role in the process of ischemic injury.
This study aims to evaluate the efficacy of the Ultrasonication-Assisted (UA) extraction on the functionality of the herbaceous biennial plant maca (Lepidium meyenii). The specific objectives include comparison of the antioxidant activities among various maca extracts, determination of the macamide B content of the extracts, and in vitro evaluation of maca on cell viability and creatine kinase (CK) activity. The antioxidant activities of the water, ethanol, and UA extracts were compared by determining the total phenolic and flavonoid contents, the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities, and the ferric reducing antioxidant power (FRAP) of the extracts. The macamide B content of maca extracts were analyzed by HPLC. The effects of the extracts on muscle cell viability and creatine kinase activity were also determined using C2C12 myoblasts. UA extraction significantly increased the total phenolic content (2.90 GAE ㎍/mg, p < 0.05), without affecting the flavonoid content. DPPH radical scavenging activity did not exhibit any statistical difference among the extracts. The ethanol and UA extracts exhibited significantly higher FRAP than the water extract (p < 0.05). The macamide B content of ethanol and UA extracts were 0.087 and 0.083 ㎍/mg, respectively. The water and UA extracts exhibited higher C2C12 muscle cell viability than the ethanol extract, and both extracts resulted in a significantly lower CK level than the H2O2-treated control group. This research suggests that the maca extract can protect muscle cells and serve as an antifatigue agent under oxidative stress conditions.
Background: Ginseng effectively reduces fatigue in both animal models and clinical trials. However, the mechanism of action is not completely understood, and its molecular targets remain largely unknown. Methods: By screening for proteins that interact with the primary components of ginseng (ginsenosides) in an affinity chromatography assay, we have identified muscle-type creatine kinase (CK-MM) as a potential target in skeletal muscle tissues. Results: Biolayer interferometry analysis showed that ginsenoside metabolites, instead of parent ginsenosides, had direct interaction with recombinant human CK-MM. Subsequently, 20(S)-protopanaxadiol (PPD), which is a ginsenoside metabolite and displayed the strongest interaction with CK-MM in the study, was selected as a representative to confirm direct binding and its biological importance. Biolayer interferometry kinetics analysis and isothermal titration calorimetry assay demonstrated that PPD specifically bound to human CK-MM. Moreover, the mutation of key amino acids predicted by molecular docking decreased the affinity between PPD and CK-MM. The direct binding activated CK-MM activity in vitro and in vivo, which increased the levels of tissue phosphocreatine and strengthened the function of the creatine kinase/phosphocreatine system in skeletal muscle, thus buffering cellular ATP, delaying exercise-induced lactate accumulation, and improving exercise performance in mice. Conclusion: Our results suggest a cellular target and an initiating molecular event by which ginseng reduces fatigue. All these findings indicate PPD as a small molecular activator of CK-MM, which can help in further developing better CK-MM activators based on the dammarane-type triterpenoid structure.
The objective of the current study was to determine the effect of pre-slaughter stress, season and breed on the activity of plasma creatine kinase (CK) and the quality of mutton. One hundred and seventy-three (173) castrated sheep from Dormer (DM), South African Mutton Merino (SAMM), Dorper (DP) and Blackhead Persian (BP) sheep breeds were used in the study. The animals were grouped according to age-groups as follows: Group 1 (6 to 8 months), Group 2 (9 to 12 months) and Group 3 (13 to 16 months). Blood samples were collected during exsanguinations using disposable vacutainer tubes for CK analysis. Representative samples of the Muscularis longissimuss thoracis et. lumborum (LTL) were collected from 84 castrated sheep, of different breeds (28 per breed) 24 h after slaughter. The following physico-chemical characteristics of mutton were determined; meat pH ($pH_{24}$), color ($L^*$, $a^*$ and $b^*$), thawing and cooking losses and Warner Braztler Shear Force (WBSF). The activity of plasma CK was significantly higher (p<0.001) in summer ($1,026.3{\pm}105.06$) and lower in winter ($723.3{\pm}77.75$). There were higher values for $L^*$ ($33.7{\pm}0.94$), $b^*$ ($11.5{\pm}0.48$) and WBSF ($29.5{\pm}1.46$) in summer season than in winter season; $L^*$ ($29.4{\pm}0.64$), $b^*$ ($10.2{\pm}0.33$) and WBSF ($21.2{\pm}0.99$). The activity of plasma CK was influenced by the type of breed with Dormer having the highest (p>0.001) levels ($1,358.6{\pm}191.08$) of CK. South African Mutton Merino had higher values for $pH_{24}$ ($5.9{\pm}0.06$), $L^*$ ($34.2{\pm}0.97$), $b^*$ ($12.2{\pm}0.50$) and WBSF ($26.8{\pm}1.51$) and Blackhead Persian had higher values ($35.5{\pm}2.17$) for cooking loss (CL%) than the other breeds. Computed Principal Component Analyses (PCA) on the activity of plasma CK and physico-chemical characteristics of mutton revealed no correlations between these variables. However, positive correlations were observed between $pH_{24}$, $L^*$, $a^*$, $b^*$, CL% and WBSF. Relationships between pre-slaughter stress, CK activity and physico-chemical characteristics of mutton were also observed. It was therefore concluded that although mutton quality and creatine kinase were not related, pre-slaughter stress, season and breed affected the activity of creatine kinase and mutton quality.
Journal of the Korean Society of Food Science and Nutrition
/
v.9
no.1
/
pp.59-73
/
1980
A detailed procedure was described for the isolation of cratine kinase (ATP-Creatine phosphotransferase, E. C. 2. 7. 3. 2.) from the muscle of the snake Bungarus fasciatus. The original isolation procedure of Kuby et al. for the rabbit muscle enzyme has been modified and extended to include a chromatographic step. The properties of the enzyme have been investigated and kinetic constants for the reverse reactions determined as the followings: 1) A molecular weight of the enzyme was determined by gel filteration on Sephadex G-100 and by electrophoresis on SDS-polyacrylamide was 86,000. 2) Two reactive sulphydryl groups were detected with dithiobis nitrobenzoic acid (DTNB). 3) The nucleotide substrate specificity in the reverse reaction was determined as ADP*2'-dADP>GDP>XDP>UDP with magnesium as the activating metal ion. 4) The order of the metal specificity in the reverse reaction Mg>Mn>$Ca{\sim}Co$ was determined with ADP as substrate. 5) A detailed kinetic analysis was carried out in the reverse direction with $MaADP^-$ as the nucleotide substrate. Initial velocity and product inhibition studies($MaADP^{2-}$ competitive with respect to MgADP- and noncompetitive with respect to $N-phosphorycreatine^{2-}$ ; Creatine competitive with respect to $N-phosphorycreatine^{2-}$ and noncompetitive with respect to Ma $ADP^-)$ indicated that the reaction obeyed a sequential mechanism of the rapid equilibrium random type.
Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang-Jin;Lee, Won Hee;Kim, Mooseong;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
Journal of Life Science
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v.31
no.3
/
pp.257-265
/
2021
Heterotrimeric kinesin-2 is a molecular motor protein of the kinesin superfamily (KIF) that moves along a microtubule plus-end directed motor protein. It consists of three different motor subunits (KIF3A, KIF3B, and KIF3C) and a kinesin-associated protein 3 (KAP3) that form a heterotrimeric complex. Heterotrimeric kinesin-2 interacts with many different binding proteins through the cargo-binding domain of the KIF3s. The activity of heterotrimeric kinesin-2 is regulated to ensure that the cargo is directed to the right place at the right time. How this regulation occurs, however, remains in question. To identify the regulatory proteins for heterotrimeric kinesin-2, we performed yeast two-hybrid screening and found a specific interaction with creatine kinase B (CKB), which is the brain isoform of cytosolic creatine kinase enzyme. CKB bound to the cargo-binding domain of KIF3A but did not interact with the KIF3B, KIF5B, or KAP3 in the yeast two-hybrid assay. The carboxyl (C)-terminal region of CKB is essential for the interaction with KIF3A. Another protein kinase, CaMKIIa, interacted with KIF3A, but GSK3a did not interact with KIF3A in the yeast two-hybrid assay. KIF3A interacted with GST-CKB-C but not with GSK-CKB-N or GST alone. When co-expressed in HEK-293T cells, CKB co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. These results suggest that the CKB-KIF3A interaction may regulate the cargo transport of heterotrimeric kinesin-2 under energy-compromised conditions in cells.
Cha, Eun Hye;Lee, Seong Jin;Lee, Ji In;Song, In Ja;Kim, Sung Chul
Journal of Acupuncture Research
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v.33
no.2
/
pp.201-210
/
2016
Objectives : The serum creatine kinase(CK) level of Amyotrophic Lateral Sclerosis(ALS) patients suggests that it may be an independent prognostic factor for the survival of ALS. We report the changes of serum CK level of ALS patients treated by administering processed Glycyrrhiza uralensis extracts and Korean medical treatments. Methods : We provided an ALS patient with processed Glycyrrhiza uralensis extracts and Korean medical treatments including acupuncture, pharmacopuncture and herbal medicine. The serum CK level was checked every month. The changes of ALSFRS-R and MRC grade were checked every month for additional result. Results : The serum CK level as the prognostic factor for the survival of ALS gradually decreased for three months. However, ALSFRS-R decreased by two points at the second trial. MRC rate had no change for three months. Conclusion : Combined administration of processed Glycyrrhiza uralensis extracts and Korean medical treatment could be meaningful possibilities in the treatment of ALS. We should conduct further studies to solve the limitations of this case study.
The role of polyamines in skeletal myoblast differentiation was investigated using the polyamine metabolic inhibitor methylglyoxal bis(guanylhydrazone)(MGBG). Concentrations of intracellular free spermidine and spermine increased 2 to 2.5-fold at the onset of myoblast fusion. The systhesis of actin, and creatine kinase activity both dramatically increased during myotube formation. However, MGBG at a concentration of 0.5 mM not only abolished the increase of intracellular free polyamines, but also reduced cell fusion to almost half the level of untreated cells, without noticeable morphological alteration. The production of actin, and creatine kinase activity were almost completely abolished by MGBG. The inhibition of myoblast fusion by MGBG was partially recovered with 0.1 mM of spermidine or spermine added externally. Results indicate that polyamines are necessary for normal myoblast differentiation. Since the first indication of myoblast differentiation is alignment of muscle cells and membrane fusion of adjacent cells, and since polyamine depletion completely inhibited the synthesis of actin, which might be associted with membranes, polyamine might be involved in myoblast differentiation through membrane reorganization events.
The Present experiment was designed to investigate the effects of Soo Jeom San on the function of heart and digestive organs. And thus it was analyzed the total acidity, recovery effect, and the other various enzyme activities such as ATPase, Creatine kinase, Aspartate transaminase, and Lactate dehydrogenase. The results were obtained as follows : 1. The Total acidity decreased after Soo JeomSan administration for 6 days, however the total acidity inoreased after the drug administration for 9 days, these phenomena demonstrate that Soo Jeom San acts as a dual factor. The mechanism of decreasing the total acidity was considered to the inhibition of ATPase activity used for HCI active transport from parietal cells. 2. Soo Jeom San recovered the islets of Langerhans which was disrupted by streptozotocin. The recovery mechanism was suggested that Soo Jeom San stimulates the ${\beta}-cell$ proliferation. 3. Soo Jeom San inhibited the enzyme activities such as Creatine kinase and Aspartate transaminase, however the drug activated Lactate dehydrogenase. According to the obtained results, Soo Jeom San may be used for curing gastric ulcer and myocardiac infarction.
Doxylamine is an antihistamine of the ethanolamine class. It is used primarily as a sleep-inducing agent. Clinicians should be aware of the complications in rhabdomyolysis patients who ingest doxylamine succinate and over-the-counter antihistamines. The easy availability of these substances increases the potential not only of intentional overdose by adults but also of inadvertent ingestion by children. Prompt intervention and careful assessment of renal function, urinary output, and serum creatine kinase levels may represent the difference between an uncomplicated and acute renal failure. Recognition of the potential for rhabdomyolysis and institution of vigorous treatment may prevent acute renal failure in patients who have taken an overdose of the drug. A 14-year-old male was found to have hematuria and oliguria. Evaluation of the patient revealed myoglobinuria, and a creatine kinase(CK) level of 117,563 IU/L. He was recovered by massive fluid administration, urine alkalization and mannitol infusion. We report a case of a suicide attempt in a child where ingestion of the doxylamine complicated by non-traumatic rhabdomyolysis with brief review related literatures.
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