• The Korean Journal of Physiology and Pharmacology
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• 제24권2호
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• pp.173-183
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• 2020

An in vitro model for ischemia/reperfusion injury has not been well-established. We hypothesized that this failure may be caused by serum deprivation, the use of glutamine-containing media, and absence of acidosis. Cell viability of H9c2 cells was significantly decreased by serum deprivation. In this condition, reperfusion damage was not observed even after simulating severe ischemia. However, when cells were cultured under 10% dialyzed FBS, cell viability was less affected compared to cells cultured under serum deprivation and reperfusion damage was observed after hypoxia for 24 h. Reperfusion damage after glucose or glutamine deprivation under hypoxia was not significantly different from that after hypoxia only. However, with both glucose and glutamine deprivation, reperfusion damage was significantly increased. After hypoxia with lactic acidosis, reperfusion damage was comparable with that after hypoxia with glucose and glutamine deprivation. Although high-passage H9c2 cells were more resistant to reperfusion damage than low-passage cells, reperfusion damage was observed especially after hypoxia and acidosis with glucose and glutamine deprivation. Cell death induced by reperfusion after hypoxia with acidosis was not prevented by apoptosis, autophagy, or necroptosis inhibitors, but significantly decreased by ferrostatin-1, a ferroptosis inhibitor, and deferoxamine, an iron chelator. These data suggested that in our SIR model, cell death due to reperfusion injury is likely to occur via ferroptosis, which is related with ischemia/reperfusion-induced cell death in vivo. In conclusion, we established an optimal reperfusion injury model, in which ferroptotic cell death occurred by hypoxia and acidosis with or without glucose/glutamine deprivation under 10% dialyzed FBS.

• Asian-Australasian Journal of Animal Sciences
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• 제19권6호
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• pp.872-876
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• 2006

The objective of this study was to evaluate Biocom (a protein source containing a high level of glutamine and alanyl-glutamine) as a replacement for glutamine (Gln) in nursery pig diets. Forty-two pigs (fourteen pigs per treatment) weaned at 28 d of age were used in a 28-d performance trial using three dietary treatments: control (no Gln), control supplemented with Gln or Biocom. The control diet was composed of corn, soybean meal, whey and fish meal. Individual body weight, pen feed disappearance and diarrhea were monitored. On d 0, 2, 7 and 14 postweaning, respectively, five pigs per treatment were selected and bled from the anterior vena cava to obtain five replicate samples of blood on each dietary treatment for determination of blood biochemical index. Dietary supplementation of Gln and Biocom did not influence performance, plasma Gln and total serum protein concentration (p>0.05). However, the addition of Gln and Biocom could prevent serum urea nitrogen and serum cortisol from increasing on d 2 postweaning (p<0.05). There were no significant differences (p>0.05) in any of the examined parameters between Gln- and Biocom-supplemented diets. In conclusion, dietary Gln did not influence the performance of early-weaned piglets owing to the complex diet containing whey, but could prevent the increase of serum urea and cortisol. Biocom could be used as a replacement for free pure Gln without any negative effect on early-weaned piglets.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.296-300
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• 1990

자외선 조사와 NTG를 처리하여 Brevibacterium flavum 10AHR(arg his $Rif^r$)과 Corynebacterium glutamicum 11TS(trp $Sm^r$의 돌연변이주를 분리하였다. B.flavum 10AHR과 C.glutamicum 11TS를 300$\mu g$/ml의 lysozyme으로 18시간 처리하여 원형질체를 형성하고, 융합시 30의 PEG 6,000으로 처리하였을 때 가장 높은$3.7\times 10^{-6}$의 융합빈도를 나타내었다.

• 한국수정란이식학회지
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• 제23권2호
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• pp.87-91
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• 2008

This study was carried out to investigate the effects of the supplementation of glutamine, glucosamine and glutathione on the porcine oocytes on IVM rates. Cocs were incubated in NCSU-23 supplemented with at $2.0{\sim}10.0\;mM$ glucosamine, $0.5{\sim}4.0\;mM$ glutamine and $0.1{\sim}1.0\;mM$ glutathione for 48 hrs. Oocytes were transferred to 50 ul drops of maturation medium covered with mineral oil and cultured in a $CO_2$ incubator ($38^{\circ}C$, 5% $CO_2$, 95% air). The IVM rates of oocytes cultured in NCSU-23 supplemented with 0.5, 1.0, 2.0 and 4.0 mM glutamine for 48 hrs were $46.0{\pm}4.5%$, $52.0{\pm}4.8%$, $50.0{\pm}4.2%$ and $44.0{\pm}4.5%$, respectively. The IVM rates of oocytes cultured in NCSU-23 supplement with 2.0, 5.0, 7.0, 10.0 mM glucosamine for 48 hrs were $44.0{\pm}4.5%$, $42.0{\pm}4.5%$, $38.0{\pm}4.6%$ and $24.0{\pm}4.8%$, respectively. The IVM rates of oocytes cultured in NCSU-23 supplemented with glucosamine were no significantly increased compare to the control ($42.5{\pm}4.0%$). The IVM rate of oocytes cultured in NCSU-23 supplemented with 3.0, 5.0, 7.0, 10.0 mM glutathione for 48 hrs were $40.0{\pm}3.2%$, $54.0{\pm}4.2%$, $48.0{\pm}4.5%$, $44.0{\pm}4.8%$, respectively. The IVM rate of oocytes cultured in NCSU-23 supplemented with glutamine and glutathione were significantly increased co~pared to those control ($42.5{\pm}4.0%$). Glucosamine did not affect the IVM rates of oocytes. IVM rates of oocytes cultured in NCSU-23 medium for 48 hrs were significantly increased compared to the cultured for 40 hrs.

• Animal Bioscience
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• 제34권12호
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• pp.1963-1973
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• 2021

Objective: The present study aimed to evaluate the influence of including L-glutamine along with glutamic acid as a supplement in weaned piglets' diets with and without whey powder. Methods: Two assays were carried out. A total of 40 piglets ([Landrace×Large White]×Pietrain) weaned at 24 days of age with an initial body weight of 6.6±0.6 kg were used in the first assay, and the following parameters were evaluated: growth performance, the incidence of diarrhea, morphometry, intestinal integrity, and hepatic glycogen index. The animals were then blocked into four groups according to different diets: diet all-grain feeding (G); diet all-grain feeding with whey powder (GW); and with vs without 1% supplementation of the commercial product containing L-glutamine and glutamic acid (A or NA). Whey powder was added according to the stage of life, corresponding to 17%, 10%, and 5%, respectively, in order to meet the need for lactose. The animals were evaluated at 24 to 42 days and at 24 to 55 days of age. The nutrient digestibility for the second assay was carried out by using 24 animals with an average weight of 11.49±1.6 kg, and the same diets were tested. Results: The supplementation of L-glutamine + glutamic acid or the addition of whey powder in diets for weaned piglets provided (p<0.05) greater feed intake, greater weight gain and improved feed conversion in the initial period (24 to 42 days age). However, in the whole period (24 to 55 days age) only amino acid supplementation affected (p<0.05) growth performance. There was a positive interaction (p<0.05) between the type of diet and L-glutamine + glutamic acid supplementation on villus height, crypt depth and the villus:crypt ratio in the duodenum. In addition, L-glutamine + glutamic acid supplementation reduced (p<0.05) the crypt depth and improved the villus:crypt ratio in the jejunum. The inclusion of whey powder affected (p<0.05) positively the digestibility coefficients analyzed except mineral matter digestibility coeficients. The supplementation of 1% the commercial product composed of L-glutamine and glutamic acid improved (p<0.05) only the digestibility coefficient of crude protein. Conclusion: These results indicate that supplementation of 1% commercial product containing L-glutamine + glutamic acid in diets for piglets from 24 to 55 days of age, dispenses with the use of whey powder when evaluating growth performance. Amino acid supplementation alone or associated with whey powder affects (p<0.05) positively the indicators of the intestinal integrity.

• 한국미생물·생명공학회지
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• 제14권6호
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• pp.495-500
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• 1986

단백질, 아미노산 등 각종 질소화합물의 공존하에서 $10^{-5}$M (0.01 $\mu$mole/$m{\ell}$)의 미량암모니아 정량이 가능한 효소적 분석법에 관하여 검토하였다. Glutamine synthetase의 L-glutamine합성 반응에서 생성되는 무기린정량법에 의하면 암모니아정량 범위는 0.01-0.10mM이였다. Glutamine synthetase와 pyruvate kinase 및 lactate dehydrogenase의 공역계를 이용하여 340nm에서의 NADH 산화에 의한 흡광도감소에 의하여 암모니아를 정량하였다. 이 방법의 정량범위는 0.01-0.05mM이었으며 반응계의 조성은 phosphoenol pyruvate, 3mM; L-glutamate, 10mM; ATP, 1mM; MgSO$_4$, 20mM;KCl, 75mM; NADH, 0.2mM; Tris-HCl buffer(pH 7.0), 100mM; pyruvate kinase, 10U/$m{\ell}$; lactate dehydrogenase, 12U/$m{\ell}$과 glutamine synthetase, 4U/$m{\ell}$이었다. 효소반응은 3$0^{\circ}C$에서 20분간 예비반응 후 각 농도의 염화암모니움을 가한후 3$0^{\circ}C$에서 30분간 반응시켰다. Glutamine synthetase와 glutamate synthase의 공역계를 사용한 암모니아정량법의 암모니아정량 범위는 0.01-0.05mM이었으며 반응계의 조성은 ATP, 5mM; L-glutamate, 5mM; $\alpha$-ketoglutarate, 5mM; MgCl$_2$, 1.5mM; NADPH, 0.15mM; Tris- HCl buffer(pH7.0) 100mM;glutamine synthetase, 1U/$m{\ell}$과 glutamate synthase, 0.5U/$m{\ell}$ 이었다. 효소반응은 3$0^{\circ}C$에서 20분간 예비반응시킨 후 각 농도의 염화암모니움을 가하여 3$0^{\circ}C$에서 30분간 반응시켰다.

• 한국토양비료학회지
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• 제3권1호
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• pp.55-59
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• 1970

대두(大豆)의 유식물(幼植物)시기에 있어서의 Glutarmine 산, Asparagine 산 및 그 Amide에 대(對)하여 각물질(各物質)의 소장(消長)을 검토하면서 대두내(大豆內)에서의 생합성(生合成)을 구명(究明)하여 보았다. 1. 산성 Amino 산으로의 Glutamine 산과 Asparagine 산의 소장곡선(消長曲線)은 자엽(子葉)에 있어서는 유식물(幼植物)시기 전반기에 있어서는 Glutamine 산, 후반기에 있어서는 Asparagin 산이 peak를 나타내어 성엽발생기를 중심(中心)으로 하여 대(對)칭적인 인상을 주었다. 그러나 배적(胚的)기관에 있어서는 양부위(兩部位)가 모두 성엽이 발생(發生)한 시기에 peak를 나타내었으며 소장곡선(消長曲線)은 거의 비슷한 증감의 현상을 나타냈다. 2. 그 Amide인 Glutamine과 Asparagine은 자엽(子葉)에나 배적(胚的)기관에서도 성엽이 발생한 시기에 peak를 나타냈으며 배적(胚的)기관에 있어서의 자엽(子葉)탈락시기에서의 감소에 비하여 자엽(子葉)에서의 급격한 증가는 대단히 인상적이다. 3. Glutamine 산과 Glutamine의 소장(消長)관계를 보면 둘다 자엽(子葉)에서 배적(胚的)기관으로 이동한 사실은 발견(發見)되었으나 그밖의 Glutamine 산은 식물(植物)의 함질소성분으로서 Ammonia를 받아드리기 위한 중요한 입구적(入口的)인 기능과 Glutamine 산 자체(自體)로서 타(他) Amino 산의 전구물질(前驅物質)인 역활을 다하고 있음을 추측할 수 있었다. 또 Glutamine의 경우는 후반기에 들어와서는 그의 가수(加水)분해로서 발생(發生)하는 Ammonia의 생성기구가 자엽(子葉)이 탈락하기 직전에는 그 기능이 상실되었음을 시사하여 주었다. 4. Asparagine 산과 Asparagine 과의 관계에서는 종자(種子) 상태(狀態)부터 감소하기 시작한 Asparagine 산은 배적(胚的)기관에 이동한 이외에도 Asparagine의 형성에 전구적(前驅的)인 물질(物質) 역할을 충분히 하여 주었다고 생각되었다. 그리고 Kreb's 회로의 생성물(生成物)로서 간주되는 Asparagine 산의 형성론(形成論)은 후반기부터 인정하게 되었다. 또 Asparagine의 함유량의 급격한 축적은 각세포(各細胞)에서 발생(發生)한 Ammonia의 농도를 유해량(有害量) 이하는 억제하기 위하여 Asparagine의 형태로 전환했다고 생각하는 설(說)과 자엽내(子葉內)에 과도(過度)하게 함유(含有)하고 있던 $NH_3$와 탄수화물로 형성 집적(集積)되어 있다는 두가지 설(說)로 적응시킬 수 있었다. 5. Amide는 발아중 또는 삼성물(森成物)에서의 경우라도 외부(外部)에서 Ammonia의 공급이 충분하면 형성량(形成量)도 많아지는데 대두내(大豆內)에 있어서는 외부(外部)에서의 공급 없이도 상당량의 Amide가 형성(形成)되었다. 이것은 대두(大豆)의 특징이 되기도 하며 특(特)히 Glutamine 보다는 Asparagine이 다량 생성되었다.

• 약학회지
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• 제23권2호
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• pp.125-128
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• 1979

In the formation of L(+) -glutamine from L(+) -glutamic acid-5-hydrazide, large amount of Raney-Ni was effective under normal pressure but hydrogenation or amonolysis of ester under pressure was useless. Preparation of glutamine with .alpha.-ketoglutaric acid (by way of 1, 4, 5, 6-tetrahydro-6-oxo-3-pyridazine carboxylic acid) is intersting but not so efficient in yield.

• 식물조직배양학회지
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• 제22권5호
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• pp.299-306
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• 1995

본 실험은 강활(O. koreanum)과 지리강활(A.purpuraefolia)의 체세포배 발생을 위한 효율적인 세포배양시스템을 확립하고자 실시하였다. 강활과 지리강활의 개화후 3주된 미숙종자는 0.1 mg/L 2,4-D와 0.1 mg/L BA 혼용처리구에서 배발생캘러스와 체세포배 발생이 가장 양호하였고, 0.1-3.0mg/L NAA가 처리된 배지에서 재분화된 식물체의 자엽과 배축의 가장자리로부터 직접 체세포배가 발생하였다. 미숙종자의 휴면을 고려하여 저온처리한 결과, 5$^{\circ}C$에서 10일동안 저장 후, 25$\pm$2$^{\circ}C$에서 배양하였을 때 반응이 없던 배양체로부터 배발생 캘러스와 체세포배가 발생하였고 이 체세포배는 줄기와 뿌리로 분화되었다. 지리강활에서는 배지내 첨가한 glutamine과 coconut milk의 뚜렷한 효과가 인정되지 않았다. 그러나 강활의 경우에는 대조구에 비하여 glutamine과 10% coconut milk가 포함된 배지에서 체세포배 발생률이 항상되었고 특히 glutamine은 coconut milk에 비해 NH$_4$NO$_3$ 대체효과가 인정되었다. 체세포배의 器內 식물체전환율은 MS 기본배지에서 89.1%로 가장 양호하였고 배발생 캘러스의 증식용 배지로는 0.01 mg/L 2,4-D와 0.01 mg/L BA혼용처리구에서 가장 좋았다.

• 미생물학회지
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• 제30권6호
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• pp.564-569
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• 1992

녹황색 세균인 Chlorobium limicola f. thiosulfatophilum NCIB 8327을 glutamate 가 질소원으로 포함된 변형 Pfennig 배지에서 배양한 다음 균체를 얻고, glutamine synthetase 효소를 초원심분리, DEAE-Sepharose CL-6B 이온교환 크로마토그래피, Sephacryl S-300 젤여과 크로마토그래피, 분취용 액체크로마토그래피의 과정을 통해 2% 수율에 46.3배의 정제배수로효소를 분리정제하였다. UV-VIS 흡수스펙트럼과 polycrylamide 젤 정기영동을 통해분리된 효소의 순수도는 확인하였다. Sephacryl S-3000 젤을 이용하여 얻은 효소의 분자량은 280 kDa 정도였으며, SDS-polyacrylamide 젤 전기영동 결과, 30 kDa 정도의 분자랴ㄹ을 갖는 단위체의 심합체로 추정된다. 이효소의 최적 온도와 pH 는 각각 $30^{\circ}C$ 와 pH 7.0 이었으나, 두기질 L-glutamine 과 hydroxylamine-HCI 에 대한 km 값은 각각 27.9 mM 과 0.92 mM 이었다. 이 효소의 활성은 alamine, glycine, tryptophan 등의 아미노산에 의해 상당한 저해를 받는 것으로 나타났으며, asparagin, lysine, leucine, valine 등에 의해서는 거의 영향을 받지 않았다.