Immobilizing engineered Escherichia coli cells into zeolitic imidazolate framework 8 for efficient biosynthesis of Ala-Gln / 生物工程学报

The α-amino acid ester acyltransferase (SAET) from Sphingobacterium siyangensis is one of the enzymes with the highest catalytic ability for the biosynthesis of l-alanyl-l-glutamine (Ala-Gln) with unprotected l-alanine methylester and l-glutamine. To improve the catalytic performance of SAET, a one-step method was used to rapidly prepare the immobilized cells (SAET@ZIF-8) in the aqueous system. The engineered Escherichia coli (E. coli) expressing SAET was encapsulated into the imidazole framework structure of metal organic zeolite (ZIF-8). Subsequently, the obtained SAET@ZIF-8 was characterized, and the catalytic activity, reusability and storage stability were also investigated. Results showed that the morphology of the prepared SAET@ZIF-8 nanoparticles was basically the same as that of the standard ZIF-8 materials reported in literature, and the introduction of cells did not significantly change the morphology of ZIF-8. After repeated use for 7 times, SAET@ZIF-8 could still retain 67% of the initial catalytic activity. Maintained at room temperature for 4 days, 50% of the original catalytic activity of SAET@ZIF-8 could be retained, indicating that SAET@ZIF-8 has good stability for reuse and storage. When used in the biosynthesis of Ala-Gln, the final concentration of Ala-Gln reached 62.83 mmol/L (13.65 g/L) after 30 min, the yield reached 0.455 g/(L·min), and the conversion rate relative to glutamine was 62.83%. All these results suggested that the preparation of SAET@ZIF-8 is an efficient strategy for the biosynthesis of Ala-Gln.

Addition of synthetic amino acids in sheep semen improves its quality after cryopreservation / Adição de aminoácidos sintéticos no sêmen ovino melhora a qualidade espermática após a criopreservação do sêmen

Some amino acids can protect mammalian sperm cells against oxidation during thermal stress caused by freezing/thawing. Thus, the objective was to evaluate the protective action of the association of the amino acids L-proline (Pro) and L-glutamine (Glu) against the cryoinjury caused to sheep sperm after cryopreservation. Eight ejaculates were collected from four sheep (n=32) and diluted in Tris-Egg Yolk-Glycerol until the final concentration of 200 x106 sptz/mL and kept in a water bath at 32 °C. The amino acids were added as follows: control (without adding amino acids), Pro+Glu 1 (100 µM Pro + 500 µM Glu), Pro+Glu 2 (300 µMPro + 1000 µM Glu), Pro+Glu 3 (500 µM Pro + 1500 µM Glu) and Pro+Glu 4 (700 µM Pro + 2000 µM Glu). Afterwards, the semen was cooled to 5 °C for 2 h, after that period, filled in 0.5 mL straws and then placed under liquid nitrogen vapor (N2L), 8 cm from the liquid sheet for 15min, and then immersed on the N2L. The samples were analyzed for sperm motility, plasma membrane and acrosomal membrane integrity, mitochondrial activity and binding test. The variables were subjected to the normality tests (Lilliefors test) and homoscedasticity tests (Cochran and Bartlett test), afterwards the variables of normal distribution were subjected to analysis of variance and the means compared by the Tukey test with a significance level of 5%. The Pro+Glu 3 group exhibited sperm with a greater (P<0.05) motility after thawing. In addition, the highest percentage of plasma and acrosomal membrane integrity were obtained using Pro+Glu 1, Pro+Glu 2 and Pro+Glu 3; and Pro+Glu 2 and Pro+Glu 3, respectively. Amino acids also kept mitochondrial activity high compared to the control, with Pro+Glu 3 resulting in greater activity (P<0.05). Sperm viability was higher (P<0.05) with the use of Pro+Glu 2 and Pro+Glu 3 than in the control. The number of sperm that showed the ability to bind to the egg yolk perivitelline membrane was higher (P<0.05) in semen treated with amino acids. It is concluded that the addition of synthetic amino acids in the semen of sheep before cryopreservation improves sperm quality and fertilization potential and can thus be added in cryopreservation protocols.

Alguns aminoácidos podem proteger as células espermáticas de mamíferos contra a oxidação durante o estresse térmico causado na congelação/descongelação. Dessa forma, objetivou-se avaliar a ação protetora da associação dos aminoácidos L-prolina (Pro) e L-glutamina (Glu) contra as crioinjúrias causadas aos espermatozoides de ovino após a criopreservação. Foram coletados oito ejaculados de quatro carneiros (n=32) e diluídos em Tris-Gema de ovo-Glicerol até a concentração final de 200 x106 sptz/mL e, mantidos em banho maria a 32 °C. Os aminoácidos foram adicionados da seguinte forma: controle (sem adição de aminoácidos), Pro+Glu 1 (100 µM Pro + 500 µM Glu), Pro+Glu 2 (300 µM Pro + 1000 µM Glu), Pro+Glu 3 (500 µM Pro + 1500 µM Glu) e Pro+Glu 4 (700 µM Pro + 2000 µM Glu). Depois, o sêmen foi resfriado a 5 °C por 2 h, após esse período, envasado em palhetas de 0,5 mL e então acondicionado sob vapor de nitrogênio líquido (N2L), a 8 cm da lâmina líquida por 15 min, e depois imersos no N2L. As amostras foram analisadas quanto à motilidade espermática, integridade da membrana plasmática e da membrana acrossomal, atividade mitocondrial e teste de ligação. As variáveis foram submetidas aos testes de normalidade (Teste de Lilliefors) e homocedacidade (Teste de Cochran e Bartlett), posteriormente as variáveis de distribuição normal foram submetidas à análise de variância e as médias comparadas pelo teste de Tukey com nível de significância de 5%. O grupo Pro+Glu 3 exibiu espermatozoides com uma maior (P<0,05) motilidade após o descongelamento. Além disso o maior percentual de integridade da membrana plasmatica e acrossomal foram obtidos utilizando Pro+Glu 1, Pro+Glu 2 e Pro+Glu 3; e Pro+Glu 2 e Pro+Glu 3, respectivamente. Os aminoácidos também mantiveram alta a atividade mitocondrial em comparação com o controle, com Pro+Glu 3 resultando numa maior atividade (P<0,05). A viabilidade dos espermatozoides foi maior (P<0,05) com o uso de Pro+Glu 2 e Pro+Glu 3 do que no controle. O número de espermatozoides que apresentaram à capacidade de ligação a membrana perivitelina da gema de ovo foi maior (P<0,05) no sêmen tratado com aminoácidos. Conclui-se que, a adição dos aminoácidos sintéticos no sêmen de ovinos antes da criopreservação melhora a qualidade espermática e o potencial fecundante, podendo assim serem adicionados em protocolos de criopreservação.

Addition of synthetic amino acids in sheep semen improves its quality after cryopreservation / Adição de aminoácidos sintéticos no sêmen ovino melhora a qualidade espermática após a criopreservação do sêmen

Some amino acids can protect mammalian sperm cells against oxidation during thermal stress caused by freezing/thawing. Thus, the objective was to evaluate the protective action of the association of the amino acids L-proline (Pro) and L-glutamine (Glu) against the cryoinjury caused to sheep sperm after cryopreservation. Eight ejaculates were collected from four sheep (n=32) and diluted in Tris-Egg Yolk-Glycerol until the final concentration of 200 x106 sptz/mL and kept in a water bath at 32 °C. The amino acids were added as follows: control (without adding amino acids), Pro+Glu 1 (100 M Pro + 500 M Glu), Pro+Glu 2 (300 M Pro + 1000 M Glu), Pro+Glu 3 (500 M Pro + 1500 M Glu) and Pro+Glu 4 (700 M Pro + 2000 M Glu). Afterwards, the semen was cooled to 5 °C for 2 h, after that period, filled in 0.5 mL straws and then placed under liquid nitrogen vapor (N2L), 8 cm from the liquid sheet for 15 min, and then immersed on the N2L. The samples were analyzed for sperm motility, plasma membrane and acrosomal membrane integrity, mitochondrial activity and binding test. The variables were subjected to the normality tests (Lilliefors test) and homoscedasticity tests (Cochran and Bartlett test), afterwards the variables of normal distribution were subjected to analysis of variance and the means compared by the Tukey test with a significance level of 5%. The Pro+Glu 3 group exhibited sp

Alguns aminoácidos podem proteger as células espermáticas de mamíferos contra a oxidação durante o estresse térmico causado na congelação/descongelação. Dessa forma, objetivou-se avaliar a ação protetora da associação dos aminoácidos L-prolina (Pro) e L-glutamina (Glu) contra as crioinjúrias causadas aos espermatozoides de ovino após a criopreservação. Foram coletados oito ejaculados de quatro carneiros (n=32) e diluídos em Tris-Gema de ovo-Glicerol até a concentração final de 200 x106 sptz/mL e, mantidos em banho maria a 32 °C. Os aminoácidos foram adicionados da seguinte forma: controle (sem adição de aminoácidos), Pro+Glu 1 (100 μM Pro + 500 μM Glu), Pro+Glu 2 (300 μM Pro + 1000 μM Glu), Pro+Glu 3 (500 μM Pro + 1500 μM Glu) e Pro+Glu 4 (700 μM Pro + 2000 μM Glu). Depois, o sêmen foi resfriado a 5 °C por 2 h, após esse período, envasado em palhetas de 0,5 mL e então acondicionado sob vapor de nitrogênio líquido (N2L), a 8 cm da lâmina líquida por 15 min, e depois imersos no N2L. As amostras foram analisadas quanto à motilidade espermática, integridade da membrana plasmática e da membrana acrossomal, atividade mitocondrial e teste de ligação. As variáveis foram submetidas aos testes de normalidade (Teste de Lilliefors) e homocedacidade (Teste de Cochran e Bartlett), posteriormente as variáveis de distribuição normal foram submetidas à análise de variância e as médias comparadas pelo teste de Tukey com nível de significância de 5%. O grupo Pro+Glu 3 exibiu espermatozoides com uma maior (P<0,05) motilidade após o descongelamento. Além disso o maior percentual de integridade da membrana plasmatica e acrossomal foram obtidos utilizando Pro+Glu 1, Pro+Glu 2 e Pro+Glu 3; e Pro+Glu 2 e Pro+Glu 3, respectivamente. Os aminoácidos também mantiveram alta a atividade mitocondrial em comparação com o controle, com Pro+Glu 3 resultando numa maior atividade (P<0,05). A viabilidade dos espermatozoides foi maior (P<0,05) com o uso de Pro+Glu 2 e Pro+Glu 3 do que no controle. O número de espermatozoides que apresentaram à capacidade de ligação a membrana perivitelina da gema de ovo foi maior (P<0,05) no sêmen tratado com aminoácidos.

Effects of glutamine and alanine supplementation, in their free form or as dipeptide, on fatigue parameters of rats submitted to resistance training / Efeitos da suplementação com glutamina e alanina, na forma livre ou como dipeptídeo, sobre parâmetros de fadiga de ratos submetidos ao treinamento resistido

Fatigue is defined as the inability to maintain muscle power and strength, impairing performance. Nutritional interventions have been used to delay this phenomenon, such as glutamine and alanine supplementation. These amino acids might attenuate several causes of fatigue, since they are important energy substrates, transport ammonia avoiding the accumulation of this toxic metabolite and attenuate muscle damage and oxidative stress. Thus, the aim of this study was to evaluate the effects of glutamine and alanine supplementation on central and muscle fatigue parameters of rats submitted to resistance training (RT). Forty adult Wistar rats (60 days) were distributed into five groups: SED (sedentary, receiving water), CON (trained, receiving water), ALA, G+A and DIP (trained and supplemented with alanine, glutamine and alanine in their free form, and Lalanyl-L-glutamine, respectively). Trained groups underwent a ladder-climbing exercise, with progressive loads, for eight weeks. Supplements were diluted in water to a 4% concentration and offered ad libitum during the last 21 days of experiment. RT increased plasma glucose, the muscle concentrations of ammonia and glutathione (GSH) and the muscle damage parameters - plasma creatine kinase (CK) and lactate dehydrogenase (LDH), whereas decreased muscle glycogen. G+A supplementation prevented the increase of muscle ammonia by RT, while ALA and G+A administration reduced plasma CK and LDH, and DIP supplementation increased the muscle content of glycogen and LDH. Contrary to expectations, DIP administration increased central fatigue parameters, such as plasma concentration of free fatty acids (FFA), hypothalamic content of serotonin and serotonin/dopamine ratio. Despite these results, there was no difference between groups in the maximum carrying capacity (MCC) tests. In conclusion, supplementation with glutamine and alanine improves some fatigue parameters, but does not affect physical performance of rats submitted to RT

O termo fadiga é definido como a incapacidade de manutenção da força e da potência musculares, prejudicando a performance. Intervenções nutricionais têm sido utilizadas para retardar este fenômeno, como a suplementação com glutamina e alanina. Estes aminoácidos poderiam atenuar diversas causas de fadiga, pois são importantes substratos energéticos, carreiam amônia evitando o acúmulo deste metabólito tóxico e atenuam a lesão muscular e o estresse oxidativo. Logo, o objetivo deste estudo foi avaliar os efeitos da suplementação com glutamina e alanina sobre parâmetros de fadiga central e muscular em ratos submetidos ao treinamento resistido (TR). Foram utilizados 40 ratos Wistar adultos (60 dias de idade), distribuídos nos grupos: SED (não treinados, recebendo água), CON (treinados, recebendo água), ALA, G+A e DIP (treinados e suplementados com alanina, glutamina e alanina livres, e L-alanil-L-glutamina, respectivamente). Os grupos treinados realizaram um exercício de escalada em escada, com aumento progressivo de carga, durante oito semanas. A suplementação foi diluída a 4% em água e ofertada via oral, ad libitum, durante os últimos 21 dias de experimento. O TR aumentou a glicemia, as concentrações musculares de amônia e de glutationa (GSH) e os parâmetros de lesão muscular - creatina quinase (CK) e lactato desidrogenase (LDH) no plasma, enquanto reduziu o glicogênio no músculo. A suplementação com G+A preveniu o aumento de amônia muscular promovido pelo TR, enquanto a administração de ALA e G+A reduziu as concentrações de CK e LDH no plasma, e a suplementação com DIP aumentou o conteúdo muscular de glicogênio e de LDH. Ao contrário do esperado, a administração de DIP aumentou parâmetros de fadiga central, como as concentrações plasmáticas de ácidos graxos livres, o conteúdo hipotalâmico de serotonina e a razão serotonina/dopamina. Apesar disso, não houve diferença entre os grupos nos testes de carga máxima. Em conclusão, a suplementação com glutamina e alanina melhora alguns parâmetros de fadiga, mas não afeta o desempenho físico em ratos submetidos ao TR

Expression, purification, characterization and application of α-amino acid ester acyltransferase from recombinant Escherichia coli / 生物工程学报

α-Amino acid ester acyltransferase (Aet) catalyzes the L-alanyl-L-glutamine forming reaction from L-alaine methylester hydrochloride and L-glutamine. In this study, the recombinant Escherichia coli saet-QC01 was used to express the α-amino acid acyltransferase, and its expression conditions were optimized. The recombinant protein was separated and purified by Ni-NTA affinity chromatography, and its enzymatic properties and catalytic applications were studied. The induction conditions suitable for enzyme production optimized were as follows: The temperature was 20 ℃, the induction stage (OD₆₀₀=2.0-2.5), IPTG concentration was 0.6 mmol/L, induction time was 12 h. The optimal reaction conditions of α-amino acid acyltransferase were 27 ℃, pH 8.5, it was most stable between pH 7.0 and 8.0 and relatively stable in an acidic environment, and low concentration of Co²⁺ or EDTA could promote the enzyme activity. Under optimal reaction conditions, 600 mmol/L of L-alaine methylester hydrochloride and 480 mmol/L of L-glutamine, the yield of L-alanyl-L-glutamine reached 78.2 g/L and productivity of 1.955 g/L/min, the conversion rate reached 75.0%. α-Amino acid ester acyltransferase has excellent acid-basei resistance, high catalytic efficiency. These characteristics suggest its application prospects in the industrial production.

Efeito da suplementação com L-glutamina e L-alanina, livres ou como dipeptídeo, sobre a lesão, inflamação e citoproteção em modelos de estresse in vivo e in vitro / Effects of supplementation with L-glutamine and L-alanine, in their free form or as dipeptide, on muscle damage, inflammation and cytoprotection of in vivo and in vitro stress models

Subprojeto 1: Determinação do efeito anti-inflamatório e citoprotetor da suplementação com L-glutamina e L-alanina, ou com L-alanil-L-glutamina (DIP) em ratos submetidos a treinamento resistido. Exercícios intensos reduzem a disponibilidade de glutamina, comprometendo a função imune e a recuperação de atletas. O objetivo do estudo foi avaliar os efeitos da suplementação oral crônica com L-glutamina e L-alanina, nas formas livres ou como dipeptídeo (DIP), sobre parâmetros de lesão, inflamação e citoproteção em ratos Wistar adultos submetidos a treinamento resistido (TR). Neste estudo, o TR reduziu a concentração de glutamina no plasma e no músculo EDL. No entanto, este efeito foi atenuado pelos suplementos contendo L-glutamina, os quais aumentaram os conteúdos da proteína de resposta ao estresse (HSP70) em células do sistema imune (PBMC) e no EDL, concomitantemente à redução da ativação do NF-kB e a da concentração de citocinas no EDL. O efeito protetor das suplementações também foi evidenciado pela atenuação de marcadores de lesão (CK e LDH) e inflamação (TNF-α e IL-1ß), bem como pelo aumento nas concentrações de marcadores anti-inflamatórios (IL-6, IL-10 e MCP-1) no plasma. Nossos resultados sugerem que a suplementação oral crônica com L-glutamina (administrada com L-alanina livre ou como DIP) promoveu efeitos citoprotetores mediados pela HSP70 em resposta à lesão e inflamação induzidas pelo TR. Subprojeto 2: Efeitos da L-alanil-L-glutamina sobre as vias de sinalização da insulina e da mTOR/S6K, e citoproteção em células musculoesqueléticas C2C12. O dipeptídeo L-alanil-L-glutamina é conhecido por modular o metabolismo e a viabilidade celular. Contudo, os efeitos sobre os componentes clássicos das vias de sinalização da insulina e da mTOR/S6K, bem como o efeito citoprotetor em células musculares, são pouco esclarecidos. O objetivo deste estudo foi investigar o efeito do DIP sobre as vias de sinalização da insulina e da mTOR/S6K em miotubos C2C12, em condições normais ou resistentes à insulina. A exposição crônica à insulina (24h) promoveu resistência à insulina, reduzindo os conteúdos totais do receptor beta (IR-ß) e do substrato do receptor de insulina (IRS-1), e diminuindo a fosforilação de IRS-1, AKT e P44/42 MAPK. Adicionalmente, houve redução na expressão do transportador de glicose (GLUT4) e HSP70, redução da viabilidade celular e menor fosforilação de p70S6k e S6, proteínas relacionadas à síntese proteica. Em contraste, a suplementação com DIP aumentou os conteúdos totais de IR-ß e IRS-1 e a fosforilação de IRS-1 e AKT. A glicólise anaeróbia e a capacidade glicolítica, além da fosforilação de p70S6k e S6, foram aumentadas pelo DIP em condições normais e na resistência à insulina. Nestas condições experimentais, nossos resultados sugerem que a suplementação com DIP melhorou as vias de sinalizações da insulina e da mTOR/S6K, aumentou a captação e metabolização da glicose, independente da estimulação com insulina e, finalmente, promoveu citoproteção resgatando parcialmente as células de um estado resistente à insulina, por meio do aumento de HSP70 e ativação das etapas finais da via mTOR/S6K.

Subproject 1: Determination of the anti-inflammatory and cytoprotective effects of supplementation with L-glutamine and L-alanine, or with L-alanyl-L-glutamine in rats submitted to resistance training. Intense exercise reduces glutamine availability, compromising immune function and recovery of athletes. The objective of the study was to evaluate the effects of chronic oral supplementation with L-glutamine and L-alanine, in their free form or as dipeptide (DIP), on muscle damage, inflammation and cytoprotection in adult Wistar rats submitted to resistance training (RT). In this study, RT reduced glutamine concentration in plasma and EDL muscle. However, this effect was attenuated by supplements containing L-glutamine, which increased the contents of the stress response protein (HSP70) in immune system cells (PBMC) and EDL, concomitantly with the reduction of NF-kB activation and the concentration of cytokines in EDL. The protective effect of supplementation was also evidenced by attenuation of lesion markers (CK and LDH) and inflammation (TNF-α and IL-1ß), as well as by the increase in anti-inflammatory plasma markers (IL-6, IL-10 and MCP-1). Our results suggest that chronic oral supplementation with L-glutamine (administered along with free L-alanine or as DIP) promoted HSP70-mediated cytoprotective effects in response to RT-induced injury and inflammation. Subproject 2: Effects of L-alanyl-L-glutamine on the components of insulin and mTOR/ S6K signaling pathways and cytoprotection in C2C12 musculoskeletal cells. The dipeptide L-alanyl-L-glutamine is known to modulate metabolism and cell viability. However, the effects on the classical components of insulin and mTOR/ S6K signaling pathways, as well as the cytoprotective effect on muscle cells, are poorly understood. The aim of this study was to investigate the effect of DIP on insulin and mTOR/ S6K signaling pathways in C2C12 myotubes, under normal or insulin resistant conditions. Chronic insulin exposure (24h) promoted insulin resistance, reducing the total contents of the insulin receptor (IR-ß) and the insulin receptor substrate (IRS-1), and decreasing the phosphorylation of IRS-1, AKT and P44/ 42 MAPK. In addition, there was a reduction in the expression of glucose transporter (GLUT4) and HSP70, reduction of cell viability and defective phosphorylation of p70S6k and S6, which are related to protein synthesis. On the other hand, DIP supplementation increased the total contents of IR-ß and IRS-1 and the phosphorylation of IRS-1 and AKT. Anaerobic glycolysis and glycolytic capacity, in addition to phosphorylation of p70S6k and S6, were increased by DIP under normal conditions and in insulin resistance. In our experimental conditions, our results suggest that DIP supplementation improved the signaling pathways of insulin and mTOR/ S6K, increased glucose uptake and metabolism, independent of insulin stimulation, and finally promoted cytoprotection by partially rescuing the cells of an insulin resistant state, by increasing HSP70 and activating the final stages of the mTOR/ S6K pathway.

Effects of L-Glutamine oral supplementation on prostate of irradiated rats

ABSTRACT Objectives To investigate the protective effect of L-Glutamine in animals undergone to ventral radiation when the target organ is not the prostate. Materials and Methods Wistar rats were divided into groups of 10 animals each: Controls (C), maintained under standard conditions and not exposed to radiation, Radiated group (R) undergone to abdominal radiation only and Radiated plus supplemented by L-glutamine group (R+G). The animals of group R+G were supplemented with L-glutamine at the beginning of the experiment until death in the 22nd day. The ventral prostate was dissected and processed for morphometrical analysis. The epithelial height, collagen density and acinar area were objectively assessed in histological sections. Results Epithelial height was significantly reduced in R group in comparison to C group (p= 0.005). However, there was no statistical difference between the C and R+G groups. Collagen surface density in the C and R groups were not statistically different, but a significant difference was observed when comparing groups R+G and R (p= 0.040). The R+G group values did not differ significantly from C group. The acinar prostate area of group R was similar to that of C (p= 0.971), but in R+G it was significantly reduced when compared with the C (p= 0.038) and R (p= 0.001) groups. Conclusions Pelvic radiation promotes structural modifications in ventral prostate of rats, which can be reduced by L-Glutamine.

Curative efficacy of L -glutamine gung shimron acid sodium in combination with esomeprazole in the treat-ment of pediatric peptic ulcer and its effects on immunological function and intestinal flora / 中国基层医药

Objective To investigate the clinical effect of L -glutamine gung shimron acid sodium in combi-nation with esomeprazole in the treatment of pediatric peptic ulcer and its effects on immunological function and intes-tinal flora.Methods 120 patients of pediatric peptic ulcer meeting the inclusion criteria were randomly divided into two groups equally.Besides conventional treatments,the control group was treated with esomeprazole,and the observa-tion group was treated with L -glutamine gung shimron acid sodium in combination with esomeprazole.After the course of treatment,the curative efficacy was compared after the eight -week follow -up.Before and after the treat-ment,T lymphocyte subpopulation (CD +4 ,CD +8 ,CD +4 /CD +8 )reflecting cellular immune function,intestinal flora and adverse reactions were compared.Results After the eight -week follow -up,the total effective rate of the observa-tion group was 96.7%,which was significantly higher than 86.7% of the control group (χ2 =3.93,P <0.05).After treatment,levels of CD +4 [(43.5 ±6.6)% vs (40.9 ±6.2)%],CD +4 /CD +8 [(1.72 ±0.38)vs (1.38 ±0.45)] were statistically higher and level of CD +8 [(26.2 ±3.7)% vs (30.1 ±5.0)%]was statistically lower in the obser-vation group in comparison with the control group (t =2.22,P <0.05;t =4.47,t =4.86,P <0.01).As to intestinal flora,the observation group had statistically higher concentration of lactobacillus [(7.5 ±1.3 )LonN /g feces vs (5.8 ±1.2)LonN /g feces],bifidobacterium[(8.3 ±1.1)LonN /g feces vs (6.6 ±1.4)LonN /g feces]in the feces in comparison with the control group after treatment (t =7.44,t =7.40,both P <0.05).During the treatment,there were no severe adverse reactions.Conclusion L -glutamine gung shimron acid sodium in combination with esome-prazole is effective for pediatric peptic ulcer,which can significantly increase curative efficacy,enhance cellular immune function and effectively regulate intestinal flora with minor adverse reactions.

Influência da L-glutamina sobre aspectos imunomodulatórios de células tronco mesenquimais medulares em situação de desnutrição proteico-energética / The influence of L-glutamine on imunumodulatory aspects of bone marrow mesenchymal stem cells under protein-energy malnutrition

A desnutrição proteico-energética (DPE) altera a hemopoese e, portanto, a geração de células imunológicas, bem como compromete o sistema imune. Desta forma, indivíduos desnutridos apresentam maior susceptibilidade a infecções. As células tronco mesenquimais (CTMs) possuem propriedades imunomodulatórias e são importantes na formação do estroma medular que sustenta a hemopoese. Visto que a L-glutamina (GLUT) é o aminoácido condicionalmente essencial mais consumido por CTMs, e que também apresenta capacidade imunomoduladora, investigou-se, neste trabalho, se a GLUT exerceria efeito sobre aspectos imunomodulatórios das CTMs em um modelo experimental de DPE. Para tanto, utilizou-se camundongos da linhagem BALB/c, os quais receberam rações normoproteica ou hipoproteica isocalóricas contendo, respectivamente, 12% e 2% de proteína por um período de 5 semanas. Após o isolamento e a caracterização de CTMs provenientes dos grupos controle (CTMct) e desnutrido (CTMdesn), cultivou-se essas células em 0, 0,6, 2 e 10mM GLUT, a fim de determinar a influência deste aminoácido sobre a expressão de fatores de transcrição e produção de citocinas por CTMct e CTMdesn. Adicionalmente, avaliou-se o efeito dos sobrenadantes das culturas de CTMct e CTMdesn sobre a proliferação e produção de citocinas por macrófagos e linfócitos esplênicos. Os animais desnutridos apresentaram anemia, leucopenia, hipoplasia medular e diminuição na concentração de proteínas séricas, albumina e préa-lbumina. A DPE não modificou a morfologia e o fenótipo das CTMs, bem como não alterou a expressão de proteínas reguladoras do ciclo celular. Por outro lado, a expressão de NFkB e STAT-3 e a produção de IL-1ß, IL-6, IL-10 e TGF-ß por CTMs foram alteradas pela DPE e variaram de acordo com as concentrações de GLUT testadas. O aumento na concentração de GLUT diminuiu a expressão de NFkB e induziu a expressão de STAT-3 por CTMs obtidas de ambos os grupos. Quanto a produção de citocinas por essas células, observou-se uma diminuição nos níveis de IL-ß e IL-6 e uma elevação nos níveis de IL-10 e TGF-ß com o aumento na concentração de GLUT. Variações na concentração desse aminoácido não alteraram a produção de IL-17 ou IFN-γ por CTMct e CTMdesn. Ademais, a concentração de GLUT alterou, de forma diretamente proporcional, a taxa de proliferação das CTMs. Os meios condicionados de CTMct e CTMdesn diminuíram a proliferação de macrófagos e linfócitos esplênicos estimulados com LPS, induziram aumento na produção da citocina antiinflamatória IL-10 por ambos os tipos celulares e diminuíram a produção das citocinas pró-inflamatórias IL-12 e TNF-α por macrófagos e IL-17 por linfócitos. Portanto, conclui-se que a GLUT possui efeito sobre a proliferação das CTMs, bem como a capacidade de imunomodular estas células

Protein-energy malnutrition (PEM) alters hemopoiesis and, therefore, the generation of immune cells, and compromises the immune system. In this way, malnourished individuals are more susceptible to infections. Mesenchymal stem cells (MSCs) have immunomodulatory properties and are important in the formation of bone marrow stroma that supports hemopoiesis. Since L-glutamine (GLUT) is a conditionally essential amino acid, which is most consumed by MSCs, and present immunomodulatory capacity, this work investigated whether GLUT would have an effect on immunomodulatory aspects of MSCs in a PEM experimental model. For this purpose, BALB/c mice were used, which received isocaloric normoproteic or hypoproteic diets, containing respectively, 12% and 2% of protein for a period of 5 weeks. After isolation and characterization of MSCs from control (MSCct) and malnourished (MSCmaln) groups, these cells were cultured with 0, 0.6, 2 and GLUT 10mM in order to determine the influence of this amino acid on the expression of transcription factors and cytokine production by MSCct and MSCmaln. Besides that, the effect of MSCct and MSCmaln culture supernatants on proliferation and cytokine production by macrophages and splenic lymphocytes was evaluated. Malnourished animals presented anemia, leucopenia, marrow hypoplasia and decreased concentration of serum proteins, albumin and prealbumin. PEM did not change morphology and phenotype of MSCs or altered the expression of cell cycle regulatory proteins. On the other hand, the expression of NFkB and STAT-3 and the production of IL-1ß, IL-6, IL-10 and TGF-ß by MSCs were modified by PEM and varied according to the tested GLUT concentrations. An increase in GLUT concentration decreased NFkB expression and induced STAT-3 expression by MSCs obtained from both groups. Regarding the production of cytokines by these cells, an increase in GLUT concentration resulted in decreased IL-1ß and IL-6 levels and increased IL- 10 and TGF-ß levels. Changes in the concentration of this aminoacid did not alter IL- 17 or IFN-γ production by MSCct and MSCmaln. Furthermore, the concentration of GLUT changed, in direct proportion, the proliferation of MSCs. The conditioned media MSCct and MSCmaln decreased the proliferation of macrophages and splenic lymphocytes stimulated with LPS, induced an increase in the production of the antiinflammatory cytokine IL-10 by both cell types, and decreased the production of proinflammatory cytokines IL-12 and TNF-α by macrophages and IL-17 by lymphocytes. Therefore, it can be concluded that GLUT has an effect on the proliferation of MSCs and it has the capacity to immunomodulate these cells

Suplementação dietética com L-glutamina e fibra solúvel na diarreia crônica refratária de paciente com aids: relato de caso / Dietary supplementation with L-glutamine and soluble fiber in refractory chronic diarrhea in aids patient: a case report

A suplementação dietética de fibras solúveis e L-glutamina tem mostrado efeitos benéficos na diarreia crônica refratária relacionada à aids. O objetivo deste trabalho foi relatar o efeito dessa suplementação em paciente com diarreia refratária associada à síndrome consumptiva do HIV/aids. A terapia nutricional enteral acrescida de L-glutamina e fibra solúvel mostrou-se eficaz no controle da diarreia, provavelmente por auxiliar no reparo da mucosa colônica,favorecendo a rápida recuperação intestinal. O resultado positivo alcançado sugere um novo protocolo coadjuvante no tratamento da diarreia crônica refratária nesses indivíduos.

Dietary supplementation of soluble fiber and L-glutamine has shown beneficial effects in refractory chronical diarrhea associated with aids. The aim of thisstudy was to report the effect of supplementation in patients with refractory diarrhea associated with wasting of HIV/aids. Enteral nutritional therapy associated with L-glutamine and soluble fiber has been effective in controlling diarrhea, probably because of its assistance in the repair of the colonic mucosa, favoring the rapid intestinal recovery. The positive result achieved suggests a new protocol support in the treatment of refractory chronical diarrhea in these individuals.

Efeitos da suplementação oral com L-arginina e L-glutamina sobre a próstata ventral de ratos submetidos à irradiação / Effects of oral supplementation with L-arginine and L-glutamine on the ventral prostate of rats subjected to irradiation

O objetivo deste trabalho foi verificar o possível efeito protetor da L-glutamina e da L-arginina sobre a próstata ventral de ratos quando administradas por gavagem. Procurou-se simular as condições clinicas de pacientes submetidos à radioterapia pélvica tendo como órgão alvo outro órgão pélvico que não a próstata. Foram analisados os efeitos desta irradiação sobre a próstata considerando este órgão como normal. Foram utilizados ratos Wistar divididos em quatro grupos: Controle, animais não submetidos à irradiação (n=10); Irradiado, submetidos à irradiação abdominal e sem suplementação adicional de aminoácido por 21 dias (n=10); Irradiado + L-glutamina, submetidos à irradiação abdominal e com suplementação adicional de L-glutamina por 21 dias (n=10); e Irradiado + L-arginina, submetidos à irradiação abdominal e com suplementação adicional de L-arginina por 21 dias (n=9). Os grupos foram mantidos em condições padrão de laboratório durante todas as etapas do experimento. Os animais submetidos à irradiação abdominal receberam uma dose única de 1000 cGy no dia 8 da experimentação. A L-glutamina e a L-arginina foram dissolvidas em água destilada e administrada por gavagem através da agulha IC-810®. As próstatas foram removidas e processadas para inclusão em parafina. Foram estudados os seguintes parâmetros: estrutura acinar (área dos ácinos e altura do epitélio) e colágeno analisados por métodos morfométricos e peso corporal. O ganho de peso nos grupos suplementados foi significativamente maior se comparado ao grupo irradiado. Houve redução da altura do epitélio no grupo irradiado quando comparado ao controle. A altura do epitélio no grupo suplementado com L-arginina foi significativamente maior do que nos grupos irradiado e suplementado com L-glutamina. Houve diminuição, de aproximadamente 18%, da área dos ácinos no grupo suplementado com L-glutamina. Já no grupo suplementado com L-arginina o valor foi similar ao do controle. O efeito da L-glutamina...

The aim of this study was to investigate the possible protective effect of L-glutamine and L-arginine on the rat ventral prostate when administered by gavage. We tried to simulate the clinical conditions of patients undergoing pelvic radiotherapy as with other pelvic organ target organ than the prostate. We analyzed the effects of irradiation on prostate considering this organ as normal. Wistar rats were divided into four groups: Control, animals not exposed to irradiation (n=10); Irradiated, submitted to abdominal irradiation and without additional amino acid supplementation (n=10); Irradiated + L-glutamine, submitted abdominal irradiation and additional supplementation with L-glutamine for 21 days (n=10), and Irradiated + L-arginine, underwent abdominal irradiation and additional supplementation with L-arginine for 21 days (n=9). The groups were kept under standard laboratory conditions during all stages of the experiment. The animals underwent abdominal irradiation received a single dose of 1000 cGy in eight days of trial. L-glutamine and L-arginine were dissolved in distilled water and administered by gavage needle through the IC-810®. The prostates were removed and processed for paraffin embedding. We studied the following parameters: acinar structure (area of acini and epithelial height) and collagen analyzed by morphometric methods and body weight. Weight gain in the supplemented groups was significantly higher compared to the irradiated group. There was a reduction in epithelial height in the irradiated group compared to control. The height of the epithelium in the group supplemented with L-arginine was significantly higher than in irradiated and supplemented with L-glutamine. There was a decrease of approximately 18% of the area of the lobes in the group supplemented with L-glutamine. In the group supplemented with L-arginine was similar to the value of control. The effect of L-glutamine on the prostatic parenchyma was to keep the collagen...

Clinical application of L-glutamine in allogeneic peripheral blood stem cell transplantation / 中国实用内科杂志

0.05).Six patients(6/23)in the Gln group developed mucositis and 11 cases(11/11)in the standard group(P

Efeito da suplementação oral de l-glutamina na parede colônica de ratos submetidos à irradiação abdominal

PURPOSE: To evaluate the structural alterations of the irradiated colonic wall in rats, verifying if L-glutamine supplementation is able to prevent them. METHODS: We used 30 male adult Wistar rats, divided into three groups: I - control, II - irradiated, and III - irradiated with L-glutamine supplementation during the 14 days of the study. Control group was maintained in laboratory standard conditions while groups II and three were submitted to abdominal radiation with an only dose of 1000 cGy in the 8th day of experimentation. All the animals were submitted to laparotomy in the 15th day for resection of the colonic segment for stereological analysis. RESULTS: Group II presented total volume of colonic wall significantly smaller than control group without altering the partial volumes of each layer. Compared to groups II and III, group III exhibited maintenance of total volume of colonic wall, nearing control group. Compared to control group, animals of group III exhibited maintenance of epithelial partial volume without altering significantly epithelial surface. CONCLUSION: It is suggested that L-glutamine supplementation can be of benefit in the irradiated colonic wall in rats.

OBJETIVO: Avaliar as alterações estruturais na parede do cólon irradiado, em ratos, verificando se a suplementação de L-glutamina pode prevení-las. MÉTODOS: Foram empregados 30 ratos Wistar, machos, adultos, divididos em três grupos: I - controle, II- irradiado e III - irradiado, com suplementação de L-glutamina durante os 14 dias do estudo. O Grupo Controle foi mantido em condições-padrão de laboratório, enquanto os grupos II e III foram submetidos à irradiação abdominal, com dose única de 1000 cGy, no 8°. dia da experimentação. Todos os animais foram operados no 15°. dia, para ressecção de segmento colônico para análise estereológica. RESULTADOS: O grupo II apresentou volume total da parede colônica significativamente menor que o Grupo Controle, sem alterar os volumes parciais de cada camada histológica. No grupo III, houve manutenção do volume total da parede do cólon, próxima ao Grupo Controle, com aumento significativo da camada mucosa, quando comparada aos grupos I e II. Na camada mucosa do grupo III, houve a manutenção do volume parcial do epitélio, comparado ao Grupo Controle, sem melhora significativa da superfície epitelial. CONCLUSÃO: Sugere-se que a suplementação de L-glutamina seja benéfica na parede do cólon irradiado, em ratos.

Distributions and functions of glutamine transporters in IEC-6 cell line / 医学研究生学报

Objective:Glutamine is the main oxidative fuel of the enterocyte which enters the enterocyte primarily via amino acid transporters.The aim of the test was to study the distributions and functions of glutamine transporters in IEC-6 cell line.Methods:The rat intestinal epithelial cell line(IEC-6) was incubated in vitro.The mRNA expression of different glutamine transporters,protein expression of system ASCT2,and the [3H]-L-glutamine uptake were measured.Results:The mRNA of system ASCT2,SN1,ATA1,LAT1,LAT2 was expressed and the protein expression of ASCT2 was also validated in IEC-6.In Na+-containing buffer,the velocity of Na+-dependent glutamine uptake was(164.07?37.94) fmol/(mg protein?10min).In Na+-free buffer,the velocity of glutamine uptake was(58.71?10.51)fmol/(mg protein?10min).With the saturate dosage of MeAIB,the velocity of glutamine uptake was(81.02 ?19.59) fmol/(mg protein?10min).Conclusion:There may be five kinds of glutamine transporters(ASCT2,SN1,ATA1,LAT1,and LAT2) in IEC-6 cell.The Na+-dependent transporter was the major contributor(64.22%) to glutamine total uptake in IEC-6.The contributions of system A and the remainder were 50.62% and 13.60%,respectively.The Na+-independent transporter was the lesser contributor(35.78%).

Effect of L-glutamine on liver Bcl-2 mRNA expression after total hepatic inflow occlusion in rats / 中国现代普通外科进展

Objective:To explore the effect of L-glutamine(Gln) on liver Bcl-2mRNA expres sion and apoptosis after total hepatic inflow occlusion in rats.Methods:Male Wistar rats were assigned randomly to three groups(n=40):Group A sham-operation group,group B control group,rats were pretreated with 4 ml 0.9% saline intraperitonally twice per day on 5 consecutive days,group C,rats were pretreated with Gln dissolved in 4 ml 0.9% saline intraperitoneally twice per day on 5 consecutive days.The rats from group B and C underwent total hepatic inflow occlusion for 35min by the pringle's manoeuvre.Ten rats from each group were randomly chosen and killed before the initiation of occlusion at 2 h,4 h,24 h after reperfusion respectively.The levels of MDA,GSH in liver tissue were measured.The serum concentrations of ALT,AST,LDH were assayed on a standard biochemistry autoanalyser.The expression levels of Bcl-2 mRNA in liver were assessed by RT-PCR.The apoptosis of liver were observed by DUTP method.The percentage of apoptosis was analyzed.Results:Compared with group B,the levels of GSH in group C increased after reperfusion(P

Protection of Glycyl-L-Glutamine against myocardial ischemia-reperfusion injury in the isolated rat heart / 中国病理生理杂志

AIM:To study the protection of Glycyl-L-Glutamine(Gly-Gln) against myocardial ischemia/reperfusion(I/R) injury in the isolated rat heart.METHODS:A model of myocardial ischemia-reperfusion injury was established with a Langendorff apparatus.Thirty male SD rats were randomly divided into four groups:control group,Gly-Gln group,I/R group and I/R+Gly-Gln group.Both I/R and I/R+Gly-Gln group were pre-perfused for 30 min,followed by 20 min ischemia and 40 min reperfusion.During reperfusion I/R+Gly-Gln group was perfused with Gly-Gln perfusate.Control group was kept perfused for 90 min.Gly-Gln group Gly-Gln perfusate was also kept perfused for 90 min.The left ventricular end-diastolic pressure(LVEDP),left ventricular developed pressure(LVDP),?dp/dtmax,heart rate(HR),monophasic action potentials(MAP) was measured during perfusion.The coronary effluent fluid was collected at different certain times.The activities of lactic dehydrogenase(LDH) and creatine kinase(CK) were determined.RESULTS:The isolated rat heart function decreased severely after 20 min ischemia and 40 min reperfusion(I/R):the LVEDP increased and the LVDP,?dp/dtmax decreased.But the LVEDP decreased and the LVDP,?dp/dtmax increased in I/R+Gly-Gln group compared with I/R group.Moreover,the activities of LDH and CK in the coronary effluent fluid decreased remarkably in I/R+Gly-Gln group compared with I/R group.CONCLUSION:Gly-Gln can play a protective role against myocardial I/R injury in isolated rat hearts via maintaining the left ventricular function and decreasing the release of LDH and CK.

Effects of l-glutamine monofluorophosphate and alendronate sodium on trabecular bone in ovariectomized rats / 中国药理学通报

AIM To evaluate whether l glutamine monofluorophosphate(MFP) together with Alendronate sodium may further increase bone mass in ovariectomized rats. METHODS Forty two 3 month old Sprague Dawley female rats were randomized into six groups:group 1 rats were sham operated(Sham),group 2 rats were ovariectomized controls(Ovx),and groups 3～6 were ovariectomized and received either 1 mg?kg -1 ?d -1 of alendronate sodium ,270 mg?kg -1 ?d -1 of calcium gluconate,5 6 mg?kg -1 ?d -1 of MFP or combination of 5 6 mg?kg -1 ?d -1 of MFP and 1 mg?kg -1 ?d -1 of alendronate sodium for 3 months. All animals received double bone fluorochrome labeling prior to sacrifice. At the end of experiment,the left tibiae were havested for histomorphometrical evaluations. RESULTS Alendronate sodium increased trabecular bone volume significantly with suppressed bone resorption and bone formation. Administrated calcium gluconate had no influence on the bone mass. MFP also could not restore cancellous bone of ovariectomized rats. Administration of both MFP and alendronate sodium increased bone mass significantly but did not increase bone mass compared with alendronate sodium. CONCLUSIONS The results indicated administration of both alendronate sodium and MFP could not further increase bone mass of ovariectomized rats.

Observation of the efficacy and safety of L-glutamine granules in treating severe burn patients / 重庆医科大学学报

Objective:To evaluate the clinical therapeutic effect of L-glutamine granules on intestinal damage of severe burn patients and the safety of the drug.Methods:Twenty-two severe burn patients were randomly divided into control group(C group, n =11) and L-glutamine treatment group ( n =11).GLN treatment group were given L-glutamine in a dose of 30g per day for 7 days and C group were given equal amount of placebo.The plasma L-glutamine concentration,the severity of intestinal mucosa damage, the level of plasma protein and parameters of immunological function were observed and the length of hospital stay was recorded.Results:After 7 days of taking L-glutamine orally,plasma L-glutamine concentration was significantly higher and the severity of intestinal mucosal damage were slighter in GLN treatment group than those in C group.The level of plasma protein and parameters of immunological function were higher in GLN treatment group than those in C group.In addition,the length of hospital stay was shorter in GLN treatment group than those in C group.Conclusion:Administration of L-glutamine can raise the plasma GLN level,abate the severity of intestinal damage obviously, maintain the barrier function of intestinal mucosa, enhance the immunological function, correct the negative nitrogen balance and promote the wound healing with no complications.