• Korean Journal of Acupuncture
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• 제31권2호
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• pp.66-78
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• 2014

목적 : 경혈의 임피던스를 측정하여 경혈의 특이성을 확보하고자 다수 연구가 진행되어왔다. 직류전압과 교류전압을 자극하여 단순히 경혈이 위치한 피부 임피던스를 측정하는 방식이 아닌 Multi-Frequency Body Impedance analysis(MF BIA) 기법을 이용하여 생체 구조 성분(세포 외액, 세포내액의 저항성분 그리고 세포막의 용량성분)을 추출하는 방법을 이용하여 경혈의 특이성을 확보하고자 한다. 인체 내 생체 이온 변화가 발생하였을 시, 경혈이 비경혈에 발생 전/후 높은 변화율이 관찰될 것이라는 가정을 하에, 생체 이온 변화를 유도하기 위하여 근피로를 유발하였으며, 유도 전/후의 생체 구조 성분을 비교 분석하였다. 방법: 대퇴직근에 근피로를 유도하기 위하여 건강한 대학생에게 Knee extension/flexion의 등속도 운동을 통하였다. 생체 이온 변화를 확인하기 위하여 젖산을 측정하였으며, 피험자마다 동일한 근피로를 유발하기 위하여 EMG(electromyogram) 분석을 통하여 peak torque와 median frequency를 분석하였다. 근피로 유발 24시간 이후까지 젖산과 peak torque와 median frequency을 측정하였으며, 각 단계마다 복토(ST32), 음시(ST33) 과 인접한 비경혈 2개에 대하여 생체 구조 성분 또한 측정하였다. 결과 : 젖산과 peak torque와 median frequency은 24시간 이후 근피로 유발 전으로 회복되었다. 세포외액 저항성분의 경우 비경혈에 비하여 복토(ST32)에서 생체 이온 변화에 따라 높은 변화율이 관찰되었으나, 음시(ST33) 에서는 비경혈에 비하여 낮은 변화율이 관찰되었다. 세포내액 저항성분은 경혈과 비경혈 사이 유의한 차이가 관찰되지 않았다. 복토(ST32)에서 세포막의 용량성분이 높은 변화율이 관찰되었지만, 음시(ST33)와 인접한 비경혈간의 뚜렷한 차이가 확인되지 않았다. 결론 : 생체 이온 변화에 따라 인접한 비경혈과 비교해보았을 시, 경혈에서의 상대적으로 높고 낮은 혹은 유사한 변화율이 관찰되었다. 따라서 경혈의 특이성을 확보하지 못하였으며, 생체 구조 성분 추출을 통하여 세포 이온 변화에 따른 경혈의 특이성을 확보하기에는 한계점을 가지고 있다고 결론을 내렸다.

• 한국융합학회논문지
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• 제8권2호
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• pp.121-126
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• 2017

본 연구에서는 피부 등가모델을 통한 세포 내액과 세포 외액 성분의 생체임피던스 특성을 파악하기 위해 개발한 휴대용 소형 임피던스 측정시스템을 이용하여 4 전극법에 의한 비 침습 방식으로 다양한 주파수 대역에서 전신의 생체전기 임피던스를 측정실험을 수행하였다. 측정은 10명의 남성 피 실험자(평균 나이 $24{\pm}3.0$세, (BMI) $20.3kg/m^2$)를 대상으로 4주간 실시하였으며, 생체임피던스 다중주파수(1 kHz, 5 kHz, 50 kHz, 70 kHz, 100 kHz, 500 kHz)대역에서 측정하였다. 실험결과, 저주파 대역인 1 kHz 대역에서 임피던스가 가장 높게 측정되었고, 고주파 대역인 500 MHz에서 임피던스가 가장 낮게 측정되었다. 특히 50 kHz 대역이상에서 임피던스가 급속히 낮아짐을 실험을 통해서 확인하였다. 또한, 피부 등가회로모델을 통한 세포 내액과 세포 외액의 임피던스 특성을 파악하는 모의실험에서도 본 연구의 생체임피던스 측정시스템의 측정값과 유사한 특성이 얻어짐을 확인할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제11권4호
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• pp.351-362
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• 1998

Microbial growth efficiency in the rumen was studied in sheep given hourly, 31.25 g oaten chaff with either 0.31 and 0.88 g urea or 1.88 and 5.63 g casein (exp. 1) and 33.33 g oaten chaff with 1.04 casein or 0.3, 0.6 and 0.9 g urea or the mixture of the casein and urea (exp. 2). Concentrations of ruminal fluid ammonia increased with increasing nitrogenous supplements. Organic matter digestibility in sacco in the rumen was not different irrespective of N sources. Isoacids and valeric acid increased with increasing ingested casein but decreased with increasing urea intake. Peptide and amino acid pools in ruminal fluid increased with increasing ammonia concentrations (exp. 2) suggesting that proteolytic activity and transportation of peptides and amino acids across microbial membrane of rumen microbes may be regulated by the metabolite mechanism (intracellular amino acids and $NH_4{^+}$, respectively). Densities of total viable and cellulolytic bacteria in ruminal fluid increased with increasing ammonia levels but that of small Entodinia decreased. The density of fungal sporangia growth on oat leaf blades decreased with increasing ammonia concentrations but appeared to remain constant in the presence of casein. Efficiency of net microbial cell synthesis was 15-28% higher when ammonia concentrations increased from 100 to above 200 mg N/l regardless of N sources. In conclusion, supplementation of preformed protein had no effect on rumen digestion and microbial growth efficiency. This could not be accounted for its effect on ruminal fluid ammonia. Increased microbial growth efficiency with increasing ammonia levels may be due to a reduction in the turnover of microbial cells within the rumen.

• Journal of Animal Science and Technology
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• 제47권3호
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• pp.363-370
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• 2005

Experiments were conducted to determine the effects of beta-mercaptoethanol ($\beta$-ME) supplements to the in vitro maturation (IVM) medium on in vitro fertilization (IVF) and intracellular glutathione (GSH) concentration. Porcine cumulus-intact oocytes were matured in TCM-I99 medium containing porcine follicular fluid, sodium pyruvate, D-glucose, FBS, hormonal supplements, and $\beta$-ME (0, 25, 50 and 100 ${\mu}$M) for 36 to 46h. After culture, cumulus-free matured oocytes were co-incubated with epididymal spermatozoa for 18h. There were no significant differences in the maturation rate among treatment groups. However, increases (P < 0.05) in intracellular GSH concentration before and after. fertilization were observed in 50 ${\mu}$M $\beta$-ME supplements to the IVM medium. Also, increases (P < 0.05) in male pronuclear formation after IVF were observed in same treatment group. In conclusion, supplementing $\beta$-ME into the IVM medium increased intracellular GSH concentrations and increased fertilization in vitro.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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• pp.269-275
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• 1998

It has been reported that the $Ca^{2+}$-ATPase and the $Ca^{2+}-Na^+$ exchanger play an important role for the regulation of intracellular $Ca^{2+}$ in somatic cells, the $Ca^{2+}$-ATPase located in the plasma membrane helps the $Ca^{2+}$ concentration in maintain low $[Ca^{2+}]_i$. Roldan & Fleming reported that the spermatozoan $Ca^{2+}$-ATPase plays an important role in the capacitation and acrosome reaction. We used to assess $Ca^{2+}$ changes by chlortetracycline (CTC) patterns in the capacitation and acrosome reaction of human and hamster spermatozoa. In the present study applying quercetin which has been known as an ATPase antagonist, the enzymatic effect of $Ca^{2+}$-ATPase on capacitation and acrosome reaction was found to be remarkable: a significant increase of the transformation from the original type to the B type and the AR type of spermatozoa. This finding suggests that $Ca^{2+}$-ATPase play an important role in the efflux and the influx of the $Ca^{2+}$ which have been known to be an essential factor for the capacitation and acrosome reaction, and that the inhibitory action of the $Ca^{2+}$-ATPase might be a prerequsit step toward the capacitation and acrosome reaction. In conclusion, this study suggest the considerable evidence as follows: the increment of the intracellular $Ca^{2+}$ concentration occurred by controlling the slope of $Ca^{2+}$ concentration through $Ca^{2+}$-ATPase activites in both the intracellular and extracellulr fluid may be important procedures for the capacitation and the acrosome reaction, and finally for fertilization of the sperm and ovum.

• The Korean Journal of Physiology and Pharmacology
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• 제22권2호
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• pp.215-223
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• 2018

Intracellular $Ca^{2+}$ mobilization is closely linked with the initiation of salivary secretion in parotid acinar cells. Reactive oxygen species (ROS) are known to be related to a variety of oxidative stress-induced cellular disorders and believed to be involved in salivary impairments. In this study, we investigated the underlying mechanism of hydrogen peroxide ($H_2O_2$) on cytosolic $Ca^{2+}$ accumulation in mouse parotid acinar cells. Intracellular $Ca^{2+}$ levels were slowly elevated when $1mM\;H_2O_2$ was perfused in the presence of normal extracellular $Ca^{2+}$. In a $Ca^{2+}-free$ medium, $1mM\;H_2O_2$ still enhanced the intracellular $Ca^{2+}$ level. $Ca^{2+}$ entry tested using manganese quenching technique was not affected by perfusion of $1mM\;H_2O_2$. On the other hand, $10mM\;H_2O_2$ induced more rapid $Ca^{2+}$ accumulation and facilitated $Ca^{2+}$ entry from extracellular fluid. $Ca^{2+}$ refill into intracellular $Ca^{2+}$ store and inositol 1,4,5-trisphosphate ($1{\mu}M$)-induced $Ca^{2+}$ release from $Ca^{2+}$ store was not affected by $1mM\;H_2O_2$ in permeabilized cells. $Ca^{2+}$ efflux through plasma membrane $Ca^{2+}-ATPase$ (PMCA) was markedly blocked by $1mM\;H_2O_2$ in thapsigargin-treated intact acinar cells. Antioxidants, either catalase or dithiothreitol, completely protected $H_2O_2-induced$ $Ca^{2+}$ accumulation through PMCA inactivation. From the above results, we suggest that excessive production of $H_2O_2$ under pathological conditions may lead to cytosolic $Ca^{2+}$ accumulation and that the primary mechanism of $H_2O_2-induced$ $Ca^{2+}$ accumulation is likely to inhibit $Ca^{2+}$ efflux through PMCA rather than mobilize $Ca^{2+}$ ions from extracellular medium or intracellular stores in mouse parotid acinar cells.

• Applied Microscopy
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• 제21권1호
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• pp.1-20
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• 1991

The effects of exogenous $Ca^{2+}$ stimulation on the post-ischemic myocardial cells were studied using isolated Langendorff-perfused guinea pig hearts. At the starting point of reperfusion, Tyrode solutions, each containing 2.0mM, 4.0mM and 8.0mM $CaCl_2$ respectively, were administered for 2 minutes apart by descending, ascending, or by combined sequences followed by standard Tyrode solution containing 1.0mM $CaCl_2$. The minutes of global ischemia produced reversible but moderate to severe degree of myocardial ultrastructrual changes including focal destruction of sarcolemma, loss of nuclear matrix, clumping and margination of chromatins, mitochondrial swelling, destruction of mitochondrial cristae, shortening of sarcomeres, focal loss of myofibrils, and separation of cell junctions. In spite of reperfusion, the ultrastructure was more severely damaged and irreversible changes such as intracellular fluid accumulation, contracted sarcomeres, mitochondrial destruction, disruption of sarcolemma, loss of nuclear matrix, and separation of cell junction were observed in a large number of cells. In contrast, Tyrode-perfused $Ca^{2+}$-stimulated myocardial cells showed relatively well preserved ultrastucture, except slight changes including focal mitochondrial swelling, widening of T-tubule, and widening of cell junctions, especially at fasciae adherentes. The post-ischemic $Ca^{2+}$-stimulated reperfused myocardial cells produced focal changes such as mitochondrial destruction, disintegration of sarcolemma, widening of T-tubule, and intracellular fluid accumulation with slight variation in degree of changes by the method of $Ca^{2+}$ administration sequence. However, in a large number of the myocardial cells, chromatins were redistributed relatively evenly in the nuclear matrix, mitochondrial cristae were tightly packed, and a considerable number of intramitochondrial granules and glycogen granules reap-pealed. These results indicate that exogenous $Ca^{2+}$ stimulation in the initial period of reperfusion may be beneficial to salvage or to reduce the post-ischemic myocardium from further deleterious changes, and that the beneficial effects may be derived from the reserves of the function of the intracellular $Ca^{2+}$ regulating organelles and/or from the responsiveness of contractile apparatus to $Ca^{2+}$ stimulation.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권5호
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• pp.424-430
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• 2005

The oral cavity is humid environment mainly due to the continuous salivary flow. The reaction of oral mucosa to fluid flow is important for homeostasis and pathogenesis. The objective of this study is the screening the change of gene expression after the application of fluid induced shear stress (FISS) on the gingival fibroblast using cDNA microarray assay. The immortalized human gingival fibroblasts were grown and FISS was applied using a cone viscometer at a rotational velocity of 40 rpm, respectively for periods of 2 and 4 hours. The synthesis of cDNA was done from the extracted total RNA and cDNA microarray assay was done subsequently. The genes that showed over 1.6 in the Cy3/Cy5 or the Cy5/Cy3 value were regarded as genes influenced significantly by the FISS application ion (/M/>0.7). The " RUNX-1" was increased its expression in 2 hours group and " RUN and SH3 domain containing 1" was increased its expression in 4 hours group. The "CC020415", "cyclin L1", "interferon regulatory factor1", "early growth response 1", "immediate early response 2", and "immediate early response 3" genes were increased their expression in 2 and 4 hours after FISS application. In conclusion, we could find many genes that were probably related to the FISS application. Interestingly, most of them were placed in similar molecular pathways and these findings improve the reliability of chip data and usefulness in overall screening. From this experiment, we could find many items for further study and it will make improvement in the understanding of intracellular events in response to FISS.

• 대한소아치과학회지
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• 제26권2호
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• pp.399-415
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• 1999

세포내 유리칼슘(free calcium, $Ca^{2+}$)은 세균에서 고등동물에 이르기까지 거의 모든 세포에서 세포 고유작용을 조절하는 중요한 세포내 신호전달체계(signal transduction system)의 매개체이다. 타액선 세포에서 부교감 신경 자극으로 타액분비가 증가될 때에도 세포내 $Ca^{2+}$ 농도 증가가 가장 중요한 역할을 한다. 그러나 췌장(pancreas)의 경우 세포내 $Ca^{2+}$ 이외에도 인접세포를 전기적 화학적으로 연결해주는 gap junction이 외분비 기능을 직접적으로 조절할 가능설이 제시되었다. 타액선 세포에서도 세포막에 고농도의 gap junction이 존재하고 있으며 gap junction을 통해 인접세포들이 전기적, 화학적으로 연계되어 있어 gap junction이 타액선 세포의 기능을 직접적으로 조절할 가능성을 내포하고 있다. 따라서 gap junction이 타액선의 타액분비 작용에도 중요한 역할을 하며 이러한 작용이 세포내 $Ca^{2+}$ 농도를 조절하여 이루어질 것이라는 가정하에 이를 확인하는 실험을 시행하였다. 흰쥐 악하선에서 유리되는 타액양을 측정하기 위해서 악하선으로 혈액을 공급하는 동맥에 가는 관을 삽입하여 생리 식염수를 관류하면서 타액선관을 통해 타액을 채취하였다. 세포내 $Ca^{2+}$ 농도는 분리한 악하선 acini 내에 $Ca^{2+}$ 농도 변화에 민감하게 반응하는 형광물질인 fura-2를 축적시키고 형광 분석기를 사용하여 형광강도를 측정하여 다음과 같은 결과를 얻었다. 1. CCh 투여로 타액 분비가 증가하였을 때 gap junction을 봉쇄하는 약물인 octanol(1 mM)을 투여하면 타액분비가 봉쇄되었으며 이는 가역적 반응이었다. 2. CCh투여로 세포내 $Ca^{2+}$ 농도가 증가하였을 때 1mM octanol을 투여하면 세포내 $Ca^{2+}$ 농도가 CCh 투여전의 상태로 감소되었다. 3. Octanol은 CCh에 의하여 유발된 초기 $Ca^{2+}$ 증가를 억제하지는 못한 반면에 후기 $Ca^{2+}$ 농도를 감소시켰다. 4. 세포막 $Ca^{2+}$ 통로를 열어주는 약물인 thapsigargin($1{\mu}M$)을 투여하여 세포내 $Ca^{2+}$ 농도를 증가시킨 후 1mM octanol을 투여하면 세포내 $Ca^{2+}$ 농도가 thapsigargin 투여 전의 상태로 감소하였다. 5. 2,5-di-tert-butyl-1,4-benzohydroquinone(TBQ)의 투여로 세포막을 통한 $Ca^{2+}$ 농도의 주기적 변동인 $Ca^{2+}$의 oscillation이 유발되었는데, 이때 1mM octanol을 투여한 경우에 $Ca^{2+}$농도의 oscillation이 정지하여 역시 gap junction을 봉쇄하면 TBQ에 의해서 유발된 세포내 $Ca^{2+}$ 농도의 주기적 변동이 사라지고 $Ca^{2+}$ 농도의 감소가 나타남을 확인하였다. 6. Gap junction을 봉쇄하는 또 다른 약물인 glycyrrhetinic acid($100{\mu}M$)도 CCh 자극으로 인한 타액분비를 억제하였다. 이상의 결과로 미루어 gap junction은 흰쥐 악하선 세포로부터의 타액분비 조절에 중요한 역할을 하는데, 이는 gap junction이 세포막 $Ca^{2+}$ 통로를 조절함으로써 수용체 자극으로 유발된 세포내 $Ca^{2+}$ 농도 변화에 영향을 미친 결과인 것으로 추측된다.

• Korean Journal of Acupuncture
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• 제37권2호
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• pp.88-96
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• 2020

Objectives : The purpose of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Euphorbia lathyris L. extracted by supercritical CO₂. Methods : Human dermal fibroblast cells dosed with the extract from Euphorbia lathyris L. were harvested and the intracellular proteome was analyzed to examine the expression of proteins related collagen synthesis pathway, metalloproteinases (MMPs), extracellular matrix (ECM)-cell interaction, cytokines, and antioxidant enzymes by 2-dimensional gel electrophoresis. Results : Fatty acid analysis of the extract from Euphorbia lathyris L. showed oleic acid was 84% and linoleic acid was 4.1%. Antioxidative effect was about 53% by beta carotene bleaching assay. In 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis, fifteen protein changes in five mechanisms which were collagen synthesis pathway, MMPs, ECM-cell interaction, cytokines, and antioxidant enzymes were analyzed. Conclusions : This study suggests the supercritical extraction from the seed of Euphorbia lathyris L. could be used as anti-oxidant substances for pharmacopuncture.