• The Korean Journal of Physiology and Pharmacology
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• v.24 no.2
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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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• v.19 no.6
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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.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.296-300
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• 1990

For interspecific portoplast fusion, Brevibacterium flauum lOAHR (Rifr axg his) and Corynebacterium glutamicum 11TS ($Sm-r$ trp) were induced by UV and NTG treatment. The protoplast fusion frequency between E. flavum XOAHR and C. glutamicum llTS was $3.7\times 10^{-6}$ with the lysozyme treatment (300 P $\mu g$ml) for 18 hrs. Genotypes of recombinants were analized as FMM ($Rif^r\; Sm^r$), FA (Rift $Sm^r$ arg), FH ($Rif^r\; Sm^r$ his), FT ($Rif^r\; Sm^r$ trp), FAH ($Rif^r\; Sm^r$ arg trp), FAT ($Rif^r\; Sm^r$ arg trp), and FAHT ($Rif^r\; Sm^r$ arg his trp). FAH 1 produced 12 fold of glutamate production compared to parental type, E. flauum 10AHR. In glutamine productivity, it produced 2.6 fold to parental type, C. glutamicum 11TS. Production of glutamate or glutamine by recombinants was involved in the specific activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS), respectively.

• Journal of Embryo Transfer
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• v.23 no.2
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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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• v.34 no.12
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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.

• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.495-500
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• 1986

Enzymatic micro-assay methods were studied those were capable of determining ammonia down to 10$^{-5}$M(0.01 $\mu$mole/ml) in the presence of other nitrogenous compounds such as protein and amino acid. Microquantities of ammonia (0.01-0.1 $\mu$mole) could be determined indirectly by measuring phosphorous, one of the products of the enzymatic reaction catalyzed by glutamine synthetase. In this reaction, L-glutamate, ATP and ammonium chloride were used as substrates, and phosphorous was formed in propotion to the concentration of ammonium chloride In the reaction mixture. Another procedure was examined in which glutamine synthetase reaction coupled with pyruvate kinase and lactate dehydrogenase reactions was used. One mililiter of the assay mixture contained; phosphoenol pyruvate, 3 mM, L-glutamate, 10 mM; ATP, 1mM: MgSO$_4$, 20 mM: KCl, 75mM: NADH, 0.2mM: Tris-HCl buffer(pH 7.0), 100mM; pyruvate kinase, 10 U: lactate dehydrogenase, 12 U and glutamine synthetase, 4 U. After preincubation for 20 min at 3$0^{\circ}C$, NH$_4$Cl was added and the rates of NADH oxidation were followed at 340nm. The effective range of this method was proved to be from 0.01 to 0.05 $\mu$mole/$m{\ell}$. Glutamine synthetase reaction coupled with glutamate synthase reaction could also be effectively used for determining microquantities of ammonia. The one mililiter assay mixture contained; ATP, 5mM: L-glutamate, 5mM; L-ketoglutarate, 5mM; MgCl$_2$, 15mM; NADPH, 0.15mM; Tris-HCl buffer(pH 7.0); 100mM; glutamine synthetase, 1U and glutamate synthase, 0.5U. After preincubation for 20min at 3$0^{\circ}C$ NH$_4$Cl was added and the rates of NADPH oxidation were followed at 340nm. The effective range of this procedure was appeared to be from 0.01 to 0.05$\mu$mole/$m{\ell}$.

• Korean Journal of Soil Science and Fertilizer
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• v.3 no.1
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• pp.55-59
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• 1970

In an effort to determine the bio-synthesis in the soybean as investigate to the variance of each substance: Glutamic acid, Aspartic acid and its amides during the growth of younger soybean plants. 1. The variance-curve of Gultamic acid and Aspartic acid as the acidic amino acids in the cotyledons was appeared the peak the first half period at Glutamic acid and the latter half at Aspartic acid in the growth of soybeans, and was received the symmetrical impression centering around the stage of adult leaf-development. But, in the embryonic organ, it appears the peak at both part, in the developmental stage of adult leaf and also appears near phenomena of increase and decrease in the variation-curve of metabolites. 2. It's amides-Gultamine and Asparagine-appears the peak at the developmental stage of adult leaf in the both cotyledons and embryonic organ, and rapid increase in the cotyledons were very impressed compare with the decrease at fallen stage of cotyledons in the embryonic organs. 3. In the relation of variance at Glutamic acid and Aspartic acid, both substance were discovered the fact of translocation from cotyledon to embryonic organ, and Glutamic acid could supposed that bear the charges of outrider substance in other amino acid as the Glutamic acid-self and major basic function for receiving the ammonia as the nitrogen contain constituent of plant. In the case of Glutamine, formation-mechanism of ammonia which develops due to its hydrolysis in the latter period of soybean growth, suggested that was forfeit its function till instance of fallen cotyledons. 4. In the relation the Aspartie acid and Asparagine, Aspartic acid which begins to decrease from seed-state was supposed that bear sufficiently the charge of outrider substance in the formation of Asparagine other than translocated to embryonic organ from cotyledon. And, formation-theory of Aspartic acid which suppose as formational substance from Kreb's cycle were recognized from latter period of soybean growth, and then, rapid accumulation of Asparagine's amounts were supposed that adapt to two theory: Theory which consider to transformation as Asparagine state for pressing to less than noxious weight the concentration of ammonia developing from the cells, and was formate and accumulate as ammonia or carbohydrates containing excess in the cotyledons.

• YAKHAK HOEJI
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• v.23 no.2
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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.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.299-306
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• 1995

The objective of this study is to establish an efficient celll culture system for somatic embryogenesis in Ostericum koreanum and Angelica purpuraefolia. The highest frequency of embryogenic callus on immature seeds of O. konanum and A, purpuraefolia. was obtained when seeds were cultured on MS medium containing 0.1 mg/L 2,4-D and 0.1 mg/L BA. However somatic embryos were formed directly from the edge of cotyledon and hypocotyl of plant which regenerated on medium supplemented with 0.1-3.0 mg/L NAA. Immature seed explane cultured at 25$\pm$2$^{\circ}C$ after 10 days treatment at 5$^{\circ}C$ produced embryogenic callus and somatic embryos, and these differentiated into whole plane. Addition of glutamine and coconut milk to media did not enhance the frequency of somatic embryogenesis in immature seed cultures of A. purpuraefolia. However in immature seed culture of O. koreanum, the frequency of somatic embryogenesis were increased on media supplemented with glutamine and 10% coconut milk. Especially addition of glutamine to the medium substituted effect of NH$_4$N0$_3$ in constast to coconut milk. The highest frequency of conversion somatic embryos into plantlet was 89.1% on MS basal medium Embryogenic calli were grown vigorously when maintained on medium with 0.01 mg/L 2,4-D and 0.01 mg/L BA.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.564-569
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• 1992

A green sulfur bacterium, Chlorobium limicola f. thiosulfatophilum NCIB 8327, was grown in modified Pfennig's medium including glu1amate as a nitrogen source. Glutamine synthetase was isolated through a series of ultracentrifugation. DEAE-Sepharose CL-6B ion exchange chromatography. Sephacryl S-300 gel permeation chromatography, and preparative HPLC. The recovery and purification fold of the enzyme were 2% and 46.3. respectively. The isolated enzyme was homogeneous on UV-Visible spectrum and polyacrylamide gel electrophoretogram. The relative molecular mass of the native enzyme was estimated to be 280,000 by gel permeation chromatography. The enzyme consisted of ten subunits with relative similar molecular mass. 30.000. which was estimated by SDS-polyacrylamide gel electrophoresis. The optimal temperature and pH of the enzyme were $30^{\circ}C$ and 7.0. Km values were 27.9 mM for L-glutamine and 0.92 mM for hydroxylamine-HCr. The enzyme activity was inhibited by alanine. glycine. and tryptophan considerably, but was not affected by asparagine, lysine. leucine. and valine.