• Asian-Australasian Journal of Animal Sciences
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• 제6권3호
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• pp.417-422
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• 1993

This study was designed to determine the changes of ATP-related compounds, especially the concentration of IMP, and compares the relationship between IMP and flavor of pork loin chops during storage as three different storage temperatures (include $4^{\circ}C$ CF and PF). Pork loin chops were kept under $4^{\circ}C$ $-1.5{\pm}0.5^{\circ}C$ (control freezing storage) and $-3{\pm}0.5^{\circ}C$ (partially freezing storage). The changes of TBA values, ATP-related compounds, pH values, in CF stored samples were higher than PF stored samples, but it had lower TBA values than $4^{\circ}C$ storage. The IMP concentration reached their peak after 2 days in $4^{\circ}C$, 5 days in CF and 7 days in PF storage, and the ATP, ADP, AMP contents of the loin chops showed minimum, respectively. Flavor of meat sensory score for $4^{\circ}C$ stored samples were more intense (p<0.05) than CF and PF samples on day 2 of storage. However, after storage for 5 days, flavor scores for CF samples were more intense (p<0.05) than $4^{\circ}C$ and PF samples. Flavor scores for PF samples were more intense (p<0.05) than $4^{\circ}C$ and CF on day 7 of storage. As the meat with the peak of IMP contents was most preferred, it was considered that the content of IMP was related to the flavor of meat and that CF, PF had influence on the IMP content.

• Asian-Australasian Journal of Animal Sciences
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• 제24권7호
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• pp.1026-1030
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• 2011

An experiment was conducted to determine the effects of caponization on the muscle composition, ATP-related compounds, the shear values, the taste panel scores and the muscle fiber areas of Taiwan country chicken cockerels. At 10 wks of age, cockerels were divided into two groups: caponized and untreated. Birds were fed grower and finisher diets ad libitum in an eighteen-week experimental period. Results showed that capons contained significantly greater muscle fat content, less breast and thigh muscle moisture content, shear value and muscle fiber area (p<0.05) than those of intact birds. However, neither treatment groups differed significantly (p>0.05) in breast and thigh muscle protein content. Compared with the intact birds, the capons contained significantly (p<0.05) less muscle ash content in the breasts, but did not differ significantly (p>0.05) in thigh muscle ash content. The breast muscle IMP and ATP+ADP+AMP+IMP contents in the intact birds were significantly (p<0.05) higher than those in the capons. The intact birds had significantly (p<0.05) higher ATP and AMP contents than did the capons as well as significantly (p<0.05) less ADP and inosine (HxR) contents in the thigh and breast muscles. The Hypoxanthine (Hx) content of the thighs in the intact birds was significantly (p<0.05) higher than that in the capons; however, there was an adverse effect on the breast muscle Hx content. The breast muscle K value in the intact birds was significantly (p<0.05) lower than that in the capons. The capons produced significantly (p<0.05) higher taste panel scores than did the intact birds for both flavor and juiciness of thigh muscle as well as for flavor and tenderness of breast muscle.

• Asian-Australasian Journal of Animal Sciences
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• 제21권2호
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• pp.190-197
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• 2008

This study was designed to analyze boar sperm to compare motility, acrosome morphology, viability and ATP by various preservation methods between Duroc boar A with cold shock resistance sperm and Duroc boar B with cold shock sensitivity sperm. Semen volume, sperm concentration, motility and normal acrosome between Duroc boar A and B did not show any differences within 2 h after collection. There were no differences in sperm motility and normal acrosome between boar A and B at 1 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively. However, sperm motility and normal acrosome from 2 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively, were higher for boar A than boar B. The frozen-thawed sperm motility and normal acrosome were higher for boar A than boar B. The sperm viability and ATP concentration according to storage period of liquid semen at $17^{\circ}C$ and $4^{\circ}C$ were higher for boar A than boar B. Also, the sperm viability and ATP concentration of frozen-thawed semen were higher for boar A than boar B. In conclusion, we found out that the original quality of boar semen with cold shock resistance sperm played an important role.

• Molecules and Cells
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• 제20권3호
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• pp.416-422
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• 2005

We previously used Southern blot analysis to detect restriction-length polymorphisms between male fertile and cytoplasmic male sterile (CMS) cytoplasms at the coxII and atp6 loci of the mtDNA of Capsicum annuum L. Two copies of atp6 were found in each male fertile and CMS pepper lines. Interestingly, one of the copies of atp6 in CMS pepper was a 3'-truncated pseudogene. The open reading frame of the coxII gene was the same in the fertile (N-) and CMS (S-) lines. However, the nucleotide sequence in the S-cytoplasm diverged from that in the N-cytoplasm 41 bp downstream of the stop codon. To develop CMS-specific sequence-characterized amplified region (SCAR) markers, inverse PCR was performed to characterize the nucleotide sequences of the 5' and 3' flanking regions of mitochondrial atp6 and coxII from the cytoplasms of male fertile (N-) and CMS (S-) pepper plants. Based on these data, two CMS-specific SCAR markers, 607 and 708 bp long, were developed to distinguish N-cytoplasm from S-cytoplasm by PCR. The CMS-specific PCR bands were verified for 20 cultivars containing either N- or S-cytoplasm. PCR amplification of CMS-specific mitochondrial nucleotide sequences will allow quick and reliable identification of the cytoplasmic types of individual plants at the seedling stage, and assessment of the purity of $F_1$ seed lots. The strategy used in this report for identifying CMS-specific markers could be adopted for many other crops where CMS is used for F1 seed production.

• Animal Bioscience
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• 제34권12호
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• pp.1912-1920
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• 2021

Objective: This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm. Methods: The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT^® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl₂ and MgCl₂ (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA. Results: The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period. Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.108-109
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• 2013

Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

• 운동영양학회지
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• 제13권1호
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• pp.15-21
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• 2009

Ergogenic aids는 에너지를 많이 소비하는 운동선수들이나 혹은 일반 사람들일지라도 기대 수준 이상으로 운동수행능력을 개선시키거나 향상시키기 위하여 흔히 사용하는 여러 가지 물질이나 처치들을 말한다. L-carnitine은 장시간 운동을 하는 동안 근육세포의 지질산화 자극에 의해 운동수행능력 증진과 유산소능력을 증진시키며 운동 수행력 증진제로 잘 알려져 있다. 본 연구에서는, 흰쥐 심근에서 유발된 field potential에 대한 L-carnitine의 작용을 통해, 심장 세포들 사이의 네트워킹으로 유도되는 세포활성 작용 기전에 미치는 영향을 확인하였다. Field potential은 multi-channel extracellular recording system(MED64 system)으로 측정하였고, L-carnitine을 100 nM, 1 μM, 10 μM, 그리고 100 μM의 농도로 처리하여 대조군에서 측정된 전류의 크기와 비교하여 결과를 분석하였다. 본 실험의 결과 심근세포에 L-carnitine을 투여 시 field potential이 증가하였다. L-carnitine과 ATP-dependant K⁺ channel 길항제인 glibenclamide을 투여 시에는 field potential의 증가가 유지된 반면, L-carnitine과 ATP-dependent K⁺ channel 효현제인 diazoxide을 투여 시에는 증가된 field potential이 다시 감소되었다. 본 연구에서는 L-carnitine이 ATP-dependant K⁺ channel을 억제하여 field potential을 증가시키는 작용이 L-carnitine의 ergogenic aids로서의 새로운 기전으로 제시될 수 있다.

• Molecules and Cells
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• 제20권2호
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• pp.235-240
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• 2005

Extracts of pine needles (Pinus densiflora Sieb. et Zucc.) have diverse physiological and pharmacological actions. In this study we show that pine needle extract alters pacemaker currents in interstitial cells of Cajal (ICC) by modulating ATP-sensitive $K^+$ channels and that this effect is mediated by prostaglandins. In whole cell patches at $30^{\circ}C$, ICC generated spontaneous pacemaker potentials in the current clamp mode (I = 0), and inward currents (pacemaker currents) in the voltage clamp mode at a holding potential of -70 mV. Pine needle extract hyperpolarized the membrane potential, and in voltage clamp mode decreased both the frequency and amplitude of the pacemaker currents, and increased the resting currents in the outward direction. It also inhibited the pacemaker currents in a dose-dependent manner. Because the effects of pine needle extract on pacemaker currents were the same as those of pinacidil (an ATP-sensitive $K^+$ channel opener) we tested the effect of glibenclamide (an ATP-sensitive $K^+$ channels blocker) on ICC exposed to pine needle extract. The effects of pine needle extract on pacemaker currents were blocked by glibenclamide. To see whether production of prostaglandins (PGs) is involved in the inhibitory effect of pine needle extract on pacemaker currents, we tested the effects of naproxen, a non-selective cyclooxygenase (COX-1 and COX-2) inhibitor, and AH6809, a prostaglandin EP1 and EP2 receptor antagonist. Naproxen and AH6809 blocked the inhibitory effects of pine needle extract on ICC. These results indicate that pine needle extract inhibits the pacemaker currents of ICC by activating ATP-sensitive $K^+$ channels via the production of PGs.

• Journal of Microbiology and Biotechnology
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• 제30권5호
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• pp.785-792
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• 2020

ʟ-Theanine, found in green tea leaves has been shown to positively affect immunity and relaxation in humans. There have been many attempts to produce ʟ-theanine through enzymatic synthesis to overcome the limitations of traditional methods. Among the many genes coding for enzymes in the ʟ-theanine biosynthesis, glutamylmethylamide synthetase (GMAS) exhibits the greatest possibility of producing large amounts of production. Thus, GMAS from Methylovorus mays No. 9 was overexpressed in several strains including vectors with different copy numbers. BW25113(DE3) cells containing the pET24ma::gmas was selected for strains. The optimal temperature, pH, and metal ion concentration were 50℃, 7, and 5 mM MnCl₂, respectively. Additionally, ATP was found to be an important factor for producing high concentration of ʟ-theanine so several strains were tested during the reaction for ATP regeneration. Baker's yeast was found to decrease the demand for ATP most effectively. Addition of potassium phosphate source was demonstrated by producing 4-fold higher ʟ-theanine. To enhance the conversion yield, GMAS was additionally overexpressed in the system. A maximum of 198 mM ʟ-theanine was produced with 16.5 mmol/l/h productivity. The whole-cell reaction involving GMAS has greatest potential for scale-up production of ʟ-theanine.

• Biomolecules & Therapeutics
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• 제19권1호
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• pp.65-69
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• 2011

In this study, we investigated whether ambroxol significantly affects secretion, production and gene expression of mucin from cultured airway epithelial cells. Confluent primary rat tracheal surface epithelial (RTSE) cells were pretreated with adenosine triphosphate (ATP) for 5 min and then treated for 30 min with ambroxol to assess the effect on mucin secretion using ELISA. Additionally, confluent NCI-H292 cells were pretreated with ambroxol for 30 min and then stimulated with EGF or PMA for 24 h. The MUC5AC mucin gene expression and mucin protein production were measured by RT-PCR and ELISA. The results were as follows: (1) ambroxol did not significantly affect ATP-induced mucin secretion from cultured RTSE cells; (2) ambroxol inhibited the production of MUC5AC mucin protein induced by EGF and PMA in NCI-H292 cells; (3) ambroxol also inhibited the expression of MUC5AC mucin gene induced by EGF and PMA in NCI-H292 cells. This result suggests that ambroxol can inhibit the production and gene expression of MUC5AC mucin, by directly acting on human airway epithelial cells.