• Title/Summary/Keyword: intracellular pH

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Comparison of Anti-oxidative Activity in a Single Serving Size of the Commercial Coffees and Teas

  • Kim, Tae-Hun;Lee, Seulgi;Seo, Jin Woo;Bing, Sun Hye;Kim, Jong Im;Kwon, Eui-Ra;Jo, Gune-Hee;Lee, Jae-Myean;Choi, Joon Sig
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.460-469
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    • 2017
  • The aim of this work was to study the comparison of anti-oxidative activity in a single serving size of commercial coffees and teas. Commercial regular coffees and teas, including, brand regular coffees ($BC_A$, $BC_B$, $BC_C$, $BC_D$, and $BC_E$), green tea ($GT_A$, $GT_B$, $GT_C$, and $GT_D$), black tea ($BT_A$, $BT_B$, and $BT_C$), pu-erh tea ($PT_A$, $PT_B$, and $PT_C$), chamomile tea ($CT_A$, $CT_B$, and $CT_C$), peppermint tea ($P_A$, $P_B$, and $P_C$), polygonatum odoratum tea ($POT_A$, $POT_B$, and $POT_C$), and jujube tea ($JT_A$, $JT_B$, and $JT_C$) were assayed for the levels of ascorbic acid, caffeine, total content of polyphenols and flavonoids, and ability to scavenge free radicals, using two in vitro antioxidant assays. The scavenging abilities of $BC_A$ and $BC_C$ were $664.91{\pm}48.87mg$ ascorbic acid equivalent/serving size and $624.36{\pm}16.18mg$ ascorbic acid equivalent/serving size, respectively. The four beverage samples ($BC_A$, $BC_C$, $GT_D$, and $BT_A$) significantly reduced the production of reactive oxygen species (ROS) and intracellular oxidative stress induced by $H_2O_2$. These results suggest that the beverages possess significant radical scavenging ability, which may be due to the presence of antioxidants. Furthermore, the significant reducing level of ROS evidences the potential antioxidant effects of these beverages in human cells.

Influence of Increased Carbon Dioxide Concentration on the Bioluminescence and Cell Density of Marine Bacteria Vibrio fischeri (이산화탄소 농도 증가에 따른 발광미생물의 상대발광량과 밀도변화에 대한 연구)

  • Sung, Chan-Gyoung;Moom, Seong-Dae;Kim, Hye-Jin;Choi, Tae-Seob;Lee, Kyu-Tae;Lee, Jung-Suk;Kang, Seong-Gil
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.8-15
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    • 2010
  • An experiment was conducted to evaluate the biologically adverse effect of increased carbon dioxide in seawater on marine bacteria, Vibrio fischeri. We measured the bioluminescence and cell density at every 6 hours for 24 hours of the whole incubation period after exposing test microbes to a range of $CO_2$ concentration such as 380(Control), 1,000, 3,000, 10,000 and 30,000 ppm, respectively. Significant effect on relative luminescence(RLU) of V. fischeri was observed in treatments with $CO_2$ concentration higher than 3,000 ppm at t=12 h. However, the difference of RLU among treatments significantly decreased with the incubation time until t=24 h. Similar trend was observed for the variation of cell density, which was measured as optical density using spectrophotometer. The results showed that a significant relationship between $CO_2$ concentration and bioluminescence of test microbes was observed for the mean time. However, the inhibition of relative bioluminescence and also cell density could be recovered at the concentration levels higher than 3,000 ppm. The dissolved $CO_2$ can be absorbed directly by cell and it can decrease the intracellular pH. Our results implied that microbes might be adversely affected at the initial growing phase by increased $CO_2$. However, they could adapt by increasing ion transport including bicarbonate and then could make their pH back to normal level. Results of this study could be supported to understand the possible influence on marine bacteria by atmospheric increase of $CO_2$ in near future and also by released $CO_2$ during the marine $CO_2$ sequestration activity.

Enzymatic characterization of Paenibacillus amylolyticus xylanases GH10 and GH30 for xylan hydrolysis (Paenibacillus amylolyticus 유래 xylanase GH10 및 GH30의 xylan 가수분해 특성)

  • Nam, Gyeong-Hwa;Jang, Myoung-Uoon;Kim, Min-Jeong;Lee, Jung-Min;Lee, Min-Jae;Kim, Tae-Jip
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.463-470
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    • 2016
  • The enzymatic degradation of xylans is the most versatile way to obtain the high value-added functional compounds or the fermentable sugars for renewable energy. The endo-${\beta}$-xylanases are the major enzymes which hydrolyze the internal ${\beta}$-1,4-linkages of xylan backbones to produce the mixtures of xylooligosaccharides including xylobiose and xylotriose. Among them, glucuronoxylanase GH30 can exclusively hydrolyze the internal ${\beta}$-1,4-linkages of xylans decorated with methylglucuronic acid branches. In the present study, two xylanolytic enzyme (PaXN_10 and PaGuXN_30) genes were cloned from Paenibacillus amylolyticus KCTC 3005, and expressed in Escherichia coli, respectively. PaXN_10 (38.7 kDa) belongs to the endo-${\beta}$-xylanases GH10 family, while PaGuXN_30 (58.5 kDa) is a member of glucuronoxylanase GH30. They share the same optimal reaction conditions at $50^{\circ}C$ and pH 7.0. Enzymatic characterization proposed that P. amylolyticus can utilize the hardwood glucuronoarabinoxylans via the cooperative actions of xylanases GH10 and GH30. The extracellular PaGuXN_30 is secreted into the medium and hydrolyzes glucuronoarabinoxylans to release a series of aldouronic acid mixtures with a methylglucuronic acid branch. The resultant products being transported into the microbial cell are successively degraded into the smaller xylooligosaccharides by the intracellular PaXN_10, which will be utilized for the cellular metabolism.

Phagocytosis of Drug-Resistant Mycobacterium Tuberculosis by Peripheral Blood Monocytes (결핵균의 약제내성과 말초혈액단핵구의 결핵균 탐식능에 관한 연구)

  • Park, Jae-Seuk;Kim, Jae-Yeal;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.470-478
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    • 1997
  • Background : Phagocytosis is probably the first step for mycobacteria to be virulent in host because virulent strains are more readily phagocytosed by macrophage than attenuated strains. According to the traditional concept, multi-drug resistant strains have been regarded as less virulent. However, this concept has been challenged, since recent studies(reported) showed that the degree of virulence and drug-resistance is not related. The purpose of this study is to evaluate whether the phagocytic activity of M.tuberculosis by peripheral blood mononuclear cells(PBMC) is different according to drug-resistance or host factor. To evaluate this, we estimated the difference of phagocytic activity of drug-resistant and drug-sensitive M.tuberculosis and also estimated the phagocytic activity of PBMC from intractable tuberculosis patients and healthy controls. Methods : PBMC from ten intractable tuberculosis patients and twelve healthy control, and three different strains of heat-killed M.tuberculosis, ie, ADS(all drug sensitive), MDR(multi-drug resistant), and ADR(all drug resistant) were used. After incubation of various strains of M.tuberculosis with PBMC, the phagocytic activity was evaluated by estimating proportion of PBMC which have phagocytosed M.tuberculosis. Results : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains(Percentage of PBMC phagocytosed M.tuberculosis in healthy control : ADS : $32.3{\pm}2.9%$, ADR : $49.6{\pm}3.4%$, p = 0.0022, Percentage of PBMC phagocytosed M.tuberculosis in intractable tuberculosis patients : ADS : $34.9{\pm}3.6%$, ADR : $50.7{\pm}4.5%$, p = 0.0069). However, there was no difference in phagocytic activity of PBMC from healthy control and intractable tuberculosis patients. Conclusion : Drug-resistant strains of M.tuberculosis were phagocytosed easily than drug sensitive strains and host factors does not seems to influence the phagocytosis of M.tuberculosis.

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Pro-inflammatory Cytokine Expression Through NF-${\kappa}B/I{\kappa}B$ Pathway in Lung Epithelial Cells (폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로에 의한 염증매개 사이토카인의 발현)

  • Park, Gye-Young;Lee, Seung-Hee;HwangBo, Bin;Yim, Jae-Joon;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.332-342
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    • 2000
  • Background : The importance of pro-inflammatory cytokines, especially tumor necrosis factor $\alpha$ (INF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$), have been extensively documented in the generation of inflammatory lung disease. Lung epithelial cells are also actively involved in initiating and maintaining inflammation by producing pro-inflammatory mediators. Understanding the mechanism of pro-inflammatory cytokine expression in lung epithelial cells is crucial to the development of new therapeutic modalities for inflammatory lung disease. Transcription of most pro-inflammatory cytokines is dependent on the activation of NF-${\kappa}B$. However, the relationship between pro-inflammatory cytokine expression and NF-${\kappa}B/I{\kappa}B$ pathway in lung epithelial cells is not clear. Methods : BEAS-2B, A549, Na-H157, NCI-H719 cells were stimulated with IL-$1{\beta}$ or TNF-$\alpha$ at various times, and then IL-8 and TNF-$\alpha$mRNA expressions were assayed by Northern blot analysis. IL-$1{\beta}$ or TNF-$\alpha$-induced NF-${\kappa}B$ activation was assessed by the nuclear translocation of p65 NF-${\kappa}B$ subunit. The degradation of $I{\kappa}B{\alpha}$ and $I{\kappa}B{\beta}$ by IL-$1{\beta}$ or TNF-$\alpha$stimulation was assayed by Western blot analysis. The phosphorylation of $I{\kappa}B{\alpha}$ was evaluated by Western blot analysis after pre-treating cells with proteasome inhibitor followed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation. The basal level of IKK $\alpha$ expression was evaluated by Western blot analysis. Results: $I{\kappa}B{\alpha}$ and $I{\kappa}B{\alpha}$ was rapidly degraded after 5 minutes of incubation with IL-$1{\beta}$ or TNF-$\alpha$ in BEAS-2B, A549, and NCI-H157 cells. The activation of NF-${\kappa}B{\alpha}$ and the induction of IL-8 and TNF-$\alpha$ mRNA expression were observed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in these cells. In contrast, neither the changes in NF-${\kappa}B/I{\kappa}B$ pathway nor IL-8 and TNF-$\alpha$mRNA expression was induced by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in NCI-H719 cells. IL-$1{\beta}$ and TNF-$\alpha$-induced $I{\kappa}B$ phosphorylation was observed in BEAS-2B, A549, and NCI-H157 cells, but not in NCI-H719 cells. The basal level of IKK$\alpha$ expression was not different between cell. Conclusion : NF-${\kappa}B/I{\kappa}B$ pathway plays an important role in the expression of pro-inflammatory cytokine in most lung epithelial cells. The absence of the effect on NF-${\kappa}B/I{\kappa}B$ pathway in NCI-H719 cells sæms to be due to the defect in the intracellular signal transduction pathway upstream to IKK.

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Anti-inflammatory and Antioxidant Effects of Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai in RAW 264.7 Cells (조팝나무 뿌리 열수 추출물이 RAW264.7 세포에서 미치는 항산화 및 항염증 활성)

  • Sim, Mi-Ok;Lee, Hyun Joo;Jang, Ji Hun;Lee, Hyo Eun;Jung, Ho-Kyung;Kim, Tae-Muk;No, Jong hyun;Jung, Jakyun;Jung, Da Eun;Cho, Hyun-Woo
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.335-342
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    • 2017
  • Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai (SSN) has been used for the anti-inflammation in traditional folk medicine. To compare water and methanol extracts of SSN, we analyzed major components using LC IT TOF MS. The major components of hot water extract were identified as caffeic acid and p-coumaric acid, but methanol extract was not well established. However, methanol extract was detected with less polarity compounds compared to hot water extract. Next, we investigated the inhibitory effects of SSN water extract on the lipopolysaccharide (LPS)-induced inflammatory response or $H_2O_2-induced$ oxidative stress in Raw 264.7 macrophage cells. SSN strongly suppressed the production of nitric oxide in LPS-induced inflammatory response without cytotoxcity. The SSN possessed free radical scavenging activities such as DPPH ($IC_{50}=320.2{\mu}g/m{\ell}$), ABTS ($IC_{50}=124.0{\mu}g/m{\ell}$), and superoxide anion radical ($IC_{50}=122.6{\mu}g/m{\ell}$). The total phenol and flavonoid content of SSN was 56.7 mg/g, and 15.1 mg/g, respectively. Furthermore, SSN decreased the $H_2O_2-induced$ cytotoxicity by enhancing the cell viability, and SSN significantly reduced the intracellular reactive oxygen species (ROS) level. Therefore, SSN may be recommended as an effective strategy to prevent and/or treat various inflammation and ROS-induced diseases.

Antimicrobial and Antioxidant Activities of Perilla frutescens var. acuta Extract and Its Fraction and Their Component Analyses (자소엽 추출물의 항균 및 항산화 효과와 성분분석)

  • Jeong, Hyo Jin;Xuan, Song Hua;Song, Ba Reum;Lee, Sang Lae;Lee, Yun Ju;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.6
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    • pp.716-725
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    • 2018
  • In this study, antimicrobial and antioxidative activities of Perilla frutescens var. acuta were investigated with 50% ethanol and the ethyl acetate fraction and also the components were analyzed. The minimum inhibitory concentration (MIC) of the ethyl acetate fraction for both Staphylococcus aureus and Pseudomonas aeruginosa were $78{\mu}g/mL$, indicating high antimicrobial effects. The free radical scavenging activity ($FSC_{50}$) and the reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) in $Fe^{3+}-EDTA/H_2O_2$ system values of the ethyl acetate fraction were $25.90{\mu}g/mL$ and $1.40{\mu}g/mL$, respectively. After the cell damage induced by $400mJ/cm^2$ UVB irradiation, the cytoprotective effect of the ethyl acetate fraction of P. frutescens var. acuta showed the concentration dependent manner ranging from 2.0 to $16.0{\mu}g/mL$. The intracellular ROS inhibitory activity in HaCaT cells decreased to 28.6% and 40.7% for the 50% ethanol extract and ethyl acetate fraction, respectively at the concentration of $32{\mu}g/mL$. Components of rosmarinic acid, luteolin, apigenin, caffeic acid and ethyl caffeate were identified in the ethyl acetate fraction. These results suggest that the extract and fraction of P. frutescens var. acuta may be applied to the field of cosmetics as a natural material that protects the skin from an external environment by having antimicrobial and antioxidative activities.

Characterization of canine coronaviruses isolated from the dogs with diarrhea in Korea (설사증 이환견(犬)으로 부터 분리(分離)한 canine coronavirus의 성상(性狀)에 관한 연구(硏究))

  • Lee, Byung-hyung;Jun, Moo-hyung;Park, Jong-hyeon;Hwang, Eui-kyung;Huh, Won
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.517-527
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    • 1994
  • An attempt was made to isolate a causative viral agents from the fecal specimens of the diseased dogs with the gastroenteritis symptoms. Two coronavirus-like agents were isolated by serial dilution end point method and plaque assay. The isolates were characterized in terms of cytopathology, antigenicity, replication, physicochemical and morphological properties. The results obtained through the experiment were as follows; 1. Among 7 fecal specimens collected from the dogs with enteric disease, 2(28.6%)coronavirus-like agents showing typical cytopathic effects of canine coronavirus were isolated, and designated as CCV D1 and CCV D2, respectively. 2. By the cross-neutralization test and indirect immunofluoresence antibody test, the isolates were antigenically indentified as the standard CCV. The viruses were replicated only in the cytoplasm of A-72 cells. 3. The isolates showed no haemagglutinating activity against the erythrocytes from 11 kinds of animals. 4. The electron microscopic observation for the isolates showed spherical and pleomorphic features, covered with club-shaped projections on the surface. The size of particles was ranged from 70 to 150nm. 5. In one-step growth curve for the isolates in A-72 cells, maximum titers of intracellular vius was $10^{4.6}$ $TCID_{50}/0.1ml$ at 46 hrs postinoculation(pi) of CCV Dl and $10^{4.4}$ $TCID_{50}/0.1ml$ at 34 hrs pi of CCV D2. The maximum titers of extracellular virus was $10^{5.5}$ $TCID_{50}/0.1ml$ at 58 hrs pi of CCV D1 and $10^{5.8}$ $TCID_{50}/0.1ml$ at 46 hrs pi of CCV D2. 6. In physicochemical property test, the isolates were very sensitive to choroform and were found to be RNA virus. The viruses was stable at pH 3.0 for 1 hr and at $22{^{\circ}C}$ for 5 hrs. However, infectivity titers reduced remarkably by treatment with $56{^{\circ}C}$ for 10min.

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Modulated Gene Expression of Toxoplasma gondii Infected Retinal Pigment Epithelial Cell Line (ARPE-19) via PI3K/Akt or mTOR Signal Pathway

  • Zhou, Wei;Quan, Juan-Hua;Gao, Fei-Fei;Ismail, Hassan Ahmed Hassan Ahmed;Lee, Young-Ha;Cha, Guang-Ho
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.135-145
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    • 2018
  • Due to the critical location and physiological activities of the retinal pigment epithelial (RPE) cell, it is constantly subjected to contact with various infectious agents and inflammatory mediators. However, little is known about the signaling events in RPE involved in Toxoplasma gondii infection and development. The aim of the study is to screen the host mRNA transcriptional change of 3 inflammation-related gene categories, PI3K/Akt pathway regulatory components, blood vessel development factors and ROS regulators, to prove that PI3K/Akt or mTOR signaling pathway play an essential role in regulating the selected inflammation-related genes. The selected genes include PH domain and leucine- rich-repeat protein phosphatases (PHLPP), casein kinase2 (CK2), vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), glutamate-cysteine ligase (GCL), glutathione S-transferase (GST), and NAD(P)H: quinone oxidoreductase (NQO1). Using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), we found that T. gondii up-regulates PHLPP2, $CK2{\beta}$, VEGF, GCL, GST and NQO1 gene expression levels, but down-regulates PHLPP1 and PEDF mRNA transcription levels. PI3K inhibition and mTOR inhibition by specific inhibitors showed that most of these host gene expression patterns were due to activation of PI3K/Akt or mTOR pathways with some exceptional cases. Taken together, our results reveal a new molecular mechanism of these gene expression change dependent on PI3K/Akt or mTOR pathways and highlight more systematical insight of how an intracellular T. gondii can manipulate host genes to avoid host defense.

The Effect of Higenamine on the Calcium Transport of Cardiac Sarcoplasmic Reticulum (Higenamine이 심근 Sarcoplasmic Reticulum의 칼슘운반에 미치는 영향에 관한 연구)

  • Kim, Joo-Hyun;Lee, Yung-Kyoon;Kim, Hae-Won;Kim, Myung-Suk;Park, Chan-Woong;Lim, Jung-Kyoo
    • The Korean Journal of Pharmacology
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    • v.18 no.2
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    • pp.79-87
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    • 1982
  • Higenamine(dl-demethylcoclaurine, dl-1-(4-hydroxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrah-ydroisoquinoline hydrochloride), which has recently been isolated from Aconite root by Drs. Kosuge and Yokota, has known to be the main cardiotonic component of the Aconite root. The present study was undertaken to investigate the effects of Higenamine on the calcium binding and release and ATPase activity of fragmented cardiac sarcoplasmic reticulum under in vitro condition. The calcium binding and release of sarcoplasmic reticulum were measured by using the double-beam spectrophotometer and the calcium sensitive dye, murexide. In the presence of $10^{-4}{\sim}5{\times}10^{-3}M$ of Higenamine, the maximal calcium binding and the initial binding rate of porcine cardiac sarcoplasmic reticulum were inhibited dose dependently by up to 43%. However, the calcium release from cardiac sarcoplasmic reticulum, which was loaded with $Ca^{++}(50{\mu}M)$, was stimulated in dose dependent manner. When incubated in the medium of 20 mM Tris-maleate(pH 7.0), 100 mM KCl, 10 mM $MgCl_2,\;0.05mM\;CaCl_2\;and\;0.014{\sim}1\;mM\;Tris-ATP\;at\;30^{\circ}C$ in the presence of Higenamine $(10^{-4}{\sim}5{\times}10^{-3}M)$, both $Ca^{++}-and\;Mg^{++}-ATPase$ of sarcoplasmic reticulum were inhibited non-competitively by Higenamine and values of $K_i$ were 4.896 mM and 6.875 mM respectively. It is suggested from the above findings that the cardiotonic effects of Higenamine might be partially explained by the inhibition of calcium binding and the stimulation of calcium release from the sarcoplasimic reticulum which may increase the free intracellular calcium that is available in the contraction of the cardiac muscle fiber.

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