Oral glutamine dipeptide or oral glutamine free amino acid reduces burned injury progression in rats / O dipeptídeo de glutamina oral ou aminoácido livre de glutamina oral reduz a progressão da lesão por queimadura em ratos

Abstract This study was carried out to evaluate the effect of Glutamine, as a dipeptide or a free amino acid form, on the progression of burn injuries in rats. Thirty male Wistar rats were burned with a comb metal plate heated in boiling water (98 °C) for three minutes, creating four rectangular full-thickness burn areas separated by three unburned interspaces (zone of stasis) in both dorsum sides. The animals were randomized into three groups (n=10): saline solution (G1-Control) and treated groups that orally received Glutamine as dipeptide (G2-Dip) or free amino acid (G3-FreeAA). Two and seven days after burn injury, lesions were photographed for unburned interspaces necrosis evolution assessment. Seven days after injury, glutathione seric was measured and histopathological analysis was performed. By photographs, there was a significant reduction in necrosis progression in G3-Free-AA between days two and seven. Histopathological analysis at day 7 showed a significantly higher stasis zone without necrosis and a higher number of fibroblasts in G2-Dip and G3-FreeAA compared with G1-Control. Also, glutathione serum dosage was higher in G2-Dip. The plasmatic glutathione levels were higher in the G2-Dip than the G1-Control, and there was a trend to higher levels in G3-FreeAA. The reduction in histological lesions, greater production of fibroblasts, and greater amounts of glutathione may have benefited the evolution of burn necrosis, which showed greater preservation of interspaces.

Resumo Este estudo foi realizado para avaliar o efeito da Glutamina, como um dipeptídeo ou forma de aminoácido livre, na progressão de queimaduras em ratos. Trinta ratos Wistar machos foram queimados com um pente de metal aquecido em água fervente (98 °C) por três minutos, criando quatro áreas retangulares queimadas separadas por três interesespaços não queimados (zona de estase) em ambos os lados do dorso. Os animais foram randomizados em três grupos (n = 10): solução salina (G1-Controle) e grupos tratados que receberam glutamina via oral como dipeptídeo (G2-Dip) ou aminoácido livre (G3-FreeAA). Dois e sete dias após a queimadura, as lesões foram fotografadas para avaliação da evolução da necrose entre os espaços não queimados. Sete dias após a lesão, foi dosada a glutationa sérica e realizada análise histopatológica. Pelas fotografias, houve uma redução significativa na progressão da necrose no G3-Free-AA entre os dias dois e sete. A análise histopatológica no dia 7 mostrou uma zona de estase significativamente maior sem necrose e número mais elevado de fibroblastos em G2-Dip e G3-FreeAA em comparação com G1-Controle. Os níveis plasmáticos de glutationa foram maiores no G2-Dip em relação ao G1-Controle, e houve tendência a níveis mais elevados no G3-FreeAA. A redução das lesões histológicas, maior produção de fibroblastos, maior quantidade de glutationa podem ter beneficiado a evolução da necrose da queimadura, que mostrou maior preservação dos interespaços.

The role of glutamine metabolism in castration-resistant prostate cancer / 亚洲男科学杂志(英文版)

Reprogramming of metabolism is a hallmark of tumors, which has been explored for therapeutic purposes. Prostate cancer (PCa), particularly advanced and therapy-resistant PCa, displays unique metabolic properties. Targeting metabolic vulnerabilities in PCa may benefit patients who have exhausted currently available treatment options and improve clinical outcomes. Among the many nutrients, glutamine has been shown to play a central role in the metabolic reprogramming of advanced PCa. In addition to amino acid metabolism, glutamine is also widely involved in the synthesis of other macromolecules and biomasses. Targeting glutamine metabolic network by maximally inhibiting glutamine utilization in tumor cells may significantly add to treatment options for many patients. This review summarizes the metabolic landscape of PCa, with a particular focus on recent studies of how glutamine metabolism alterations affect therapeutic resistance and disease progression of PCa, and suggests novel therapeutic strategies.

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.

Suplementação de glutamina na recuperação da flora intestinal / Glutamine supplementation in gut flora recovery / La suplementación con glutamina en la recuperación de la flora intestinal

Objetivo: evidenciar através de uma revisão integrativa os resultados clínicos atuais da suplementação de glutamina na melhora da saúde intestinal, através de sintomas e exames bioquímicos. Método: Revisão integrativa da literatura realizada no período de setembro de de 2021 nas bases de dados Pubmed e Scielo. Resultados: Foi realizado uma busca pelos descritores em saúde determinados e foram selecionadas 08 produções cientificas que atenderam os critérios de inclusão. Conclusão: Sugere-se novas pesquisas que elucidem as dosagens, efeitos colaterais e respostas terapêuticas da glutamina sobre parâmetros de saúde intestinal.

Objective: to evidence through an integrative review the current clinical results of glutamine supplementation in the improvement of intestinal health, through symptoms and biochemical tests. Method: Integrative review of the literature conducted in the period of September 2021 in the Pubmed and Scielo databases. Results: A search was performed for the defined health descriptors and 08 scientific productions were selected that met the inclusion criteria. Conclusion: Further research is suggested to elucidate the dosages, side effects and therapeutic responses of glutamine on intestinal health parameters.

Objetivo: evidenciar a través de una revisión integradora los resultados clínicos actuales de la suplementación con glutamina en la mejora de la salud intestinal, a través de síntomas y pruebas bioquímicas. Método: Revisión integradora de la literatura realizada en el periodo de septiembre de 2021 en las bases de datos Pubmed y Scielo. Resultados: Se realizó una búsqueda de los descriptores de salud definidos y se seleccionaron 08 producciones científicas que cumplieron con los criterios de inclusión. Conclusión: Se sugiere investigación adicional para dilucidar las dosis, los efectos secundarios y las respuestas terapéuticas de la glutamina en los parámetros de salud intestinal.

Efeitos da nutrição parenteral ou enteral, associadas ou não à glutamina, sobre a motilidade gastrintestinal em equinos submetidos à inanição e realimentação / Effects of parenteral or enteral nutrition, associated or not with glutamine, on gastrointestinal motility in horses submmited to starvation and refeeding

Este estudo teve por objetivo avaliar os efeitos da nutrição parenteral total ou enteral, associadas ou não à glutamina, sobre a motilidade gastrintestinal em equinos submetidos à inanição e realimentação. Foram utilizados 16 equinos adultos hígidos, sem raça definida, de ambos os sexos, quatro machos e 12 fêmeas, com idade variando entre quatro e 14 anos e peso corporal médio de 248,40 + 2,28 kg, divididos em quatro grupos, quatro animais por grupo: Grupo I (ENTGL): fluidoterapia enteral com eletrólitos associada a glutamina; Grupo II (PARGL): Nutrição parenteral total (NPT) associada a glutamina; Grupo III (ENTFL): fluidoterapia enteral com eletrólitos; Grupo IV (PARFL): fluidoterapia parenteral. O delineamento experimental foi inteiramente ao acaso, em um esquema fatorial 4x12 (grupos x tempo de colheita), para cada fase, e suas médias comparadas pelo teste de Duncan ao nível de 5% de significância. Independente do grupo experimental ocorreu redução da motilidade gastrintestinal durante a fase de inanição, mais pronunciada nos grupos PARGL e PARFL. Uma vez restabelecida a alimentação a motilidade gastrintestinal retornou à normalidade.

This study aimed to evaluate the effects of enteral or total parenteral nutrition, associated or not with glutamine, on gastrointestinal motility in horses subjected to starvation and refeeding. 16 healthy, mixed-breed adult horses of both sexes, four geldings and 12 mares, with ages ranging from four to 14 years and an average body weight of 248.40 + 2.28 kg, were divided into four groups, four animals per group: Group I (ENTGL): enteral fluid therapy with electrolytes associated with glutamine; Group II (PARGL): total parenteral nutrition (TPN) associated with glutamine; Group III (ENTFL): enteral fluid therapy with electrolytes; Group IV (PARFL): parenteral fluid therapy. The experimental design was entirely randomized, in a 4x12 factorial scheme (groups x harvest time), for each phase, and their means compared by the Duncan test at the level of 5% significance. Regardless of the experimental group, there was a reduction in gastrointestinal motility during the starvation phase, which was more pronounced in the PARGL and PARFL groups. Once the food was restored, gastrointestinal motility returned to normal.

Efeitos da nutrição parenteral ou enteral, associadas ou não à glutamina, sobre a motilidade gastrintestinal em equinos submetidos à inanição e realimentação / Effects of parenteral or enteral nutrition, associated or not with glutamine, on gastrointestinal motility in horses submmited to starvation and refeeding

Este estudo teve por objetivo avaliar os efeitos da nutrição parenteral total ou enteral, associadas ou não à glutamina, sobre a motilidade gastrintestinal em equinos submetidos à inanição e realimentação. Foram utilizados 16 equinos adultos hígidos, sem raça definida, de ambos os sexos, quatro machos e 12 fêmeas, com idade variando entre quatro e 14 anos e peso corporal médio de 248,40 + 2,28 kg, divididos em quatro grupos, quatro animais por grupo: Grupo I (ENTGL): fluidoterapia enteral com eletrólitos associada a glutamina; Grupo II (PARGL): Nutrição parenteral total (NPT) associada a glutamina; Grupo III (ENTFL): fluidoterapia enteral com eletrólitos; Grupo IV (PARFL): fluidoterapia parenteral. O delineamento experimental foi inteiramente ao acaso, em um esquema fatorial 4x12 (grupos x tempo de colheita), para cada fase, e suas médias comparadas pelo teste de Duncan ao nível de 5% de significância. Independente do grupo experimental ocorreu redução da motilidade gastrintestinal durante a fase de inanição, mais pronunciada nos grupos PARGL e PARFL. Uma vez restabelecida a alimentação a motilidade gastrintestinal retornou à normalidade.

This study aimed to evaluate the effects of enteral or total parenteral nutrition, associated or not with glutamine, on gastrointestinal motility in horses subjected to starvation and refeeding. 16 healthy, mixed-breed adult horses of both sexes, four geldings and 12 mares, with ages ranging from four to 14 years and an average body weight of 248.40 + 2.28 kg, were divided into four groups, four animals per group: Group I (ENTGL): enteral fluid therapy with electrolytes associated with glutamine; Group II (PARGL): total parenteral nutrition (TPN) associated with glutamine; Group III (ENTFL): enteral fluid therapy with electrolytes; Group IV (PARFL): parenteral fluid therapy. The experimental design was entirely randomized, in a 4x12 factorial scheme (groups x harvest time), for each phase, and their means compared by the Duncan test at the level of 5% significance. Regardless of the experimental group, there was a reduction in gastrointestinal motility during the starvation phase, which was more pronounced in the PARGL and PARFL groups. Once the food was restored, gastrointestinal motility returned to normal.

Cigarette smoke promotes oral leukoplakia via regulating glutamine metabolism and M2 polarization of macrophage / 国际口腔科学杂志·英文版

Oral immunosuppression caused by smoking creates a microenvironment to promote the occurrence and development of oral mucosa precancerous lesions. This study aimed to investigate the role of metabolism and macrophage polarization in cigarette-promoting oral leukoplakia. The effects of cigarette smoke extract (CSE) on macrophage polarization and metabolism were studied in vivo and in vitro. The polarity of macrophages was detected by flow cytometric analysis and qPCR. Liquid chromatography-mass spectrometry (LC-MS) was used to perform a metabolomic analysis of Raw cells stimulated with CSE. Immunofluorescence and flow cytometry were used to detect the polarity of macrophages in the condition of glutamine abundance and deficiency. Cell Counting Kit-8 (CCK-8), wound-healing assay, and Annexin V-FITC (fluorescein isothiocyanate)/PI (propidium iodide) double-staining flow cytometry were applied to detect the growth and transferability and apoptosis of Leuk-1 cells in the supernatant of Raw cells which were stimulated with CSE, glutamine abundance and deficiency. Hyperkeratosis and dysplasia of the epithelium were evident in smoking mice. M2 macrophages increased under CSE stimulation in vivo and in vitro. In total, 162 types of metabolites were detected in the CSE group. The metabolites of nicotine, glutamate, arachidic acid, and arginine changed significantly. The significant enrichment pathways were also selected, including nicotine addiction, glutamine and glutamate metabolism, and arginine biosynthesis. The results also showed that the supernatant of Raw cells stimulated by CSE could induce excessive proliferation of Leuk-1 and inhibit apoptosis. Glutamine abundance can facilitate this process. Cigarette smoke promotes oral leukoplakia via regulating glutamine metabolism and macrophage M2 polarization.

Abnormal neurobiochemical metabolites in the first / 中南大学学报(医学版)

OBJECTIVES@#To explore the metabolite characteristics in medial prefrontal cortex (mPFC) by @*METHODS@#A total of 46 patients with the first-episode schizophrenia (FES), 49 people with clinical high risk (CHR), 61 people with genetic high risk (GHR), and 58 healthy controls (HC) were enrolled. The levels of N-acetylaspartylglutamate+N-acetylaspartate (tNAA), choline-containing compounds (Cho) and myo-inositol (MI), glutamate+glutamine (Glx) in medial prefrontal cortex were measured by single-voxel @*RESULTS@#There were significant differences in Glx, tNAA, and MI concentrations among 4 groups (all @*CONCLUSIONS@#The decreased levels of MI and Glx in the FES patients suggest that there may be glial functional damage and glutamatergic transmitter dysfunction in the early stage of the disease. The compensatory increase of metabolites may be a protective factor for schizophrenia in the genetic individuals.

Protective Effect of Naoxintong Capsule () Combined with Guhong Injection () on Rat Brain Microvascular Endothelial Cells during Cerebral Ischemia-Reperfusion Injury / 中国结合医学杂志

OBJECTIVE@#To investigate the synergistic effect of Naoxintong Capsule (NXTC, ) and Guhong Injection (GHI, ) on cerebral ischemia-reperfusion (I/R) injury.@*METHODS@#Forty-eight Sprague-Dawley rats were divided into 6 groups: control group, oxygen and glucose deprivation (OGD) group, nimodipine group (9.375 mg/kg), NXTC group (0.5 g/kg), GHI group (5 mL/kg) and NXTC+GHI group (0.5 g/kg NXTC+5 mL/kg GHI), after the onset of reperfusion and once per day for the following 7 days. Blood was collected 1 h after final administration, and the sera were collected. Cultured primary rat brain microvascular endothelial cells (rBMECs) were subjected to OGD to establish a cell injury model. Untreated rBMECs were used as blank control. The cell counting kit-8 assay was used to assess cell viability using the sera. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were assessed using an enzyme-linked immunosorbent assay. Apoptosis was evaluated after Hoechst33342 staining using fluorescence microscopy and flow cytometry. JC-1 staining was performed to assess changes in mitochondrial membrane potential.@*RESULTS@#Statistical analysis indicated that more than 95% of the cells were rBMECs. Compared with the OGD group, the cellular morphology of the all drug delivery groups improved. In particular, the combined drug group had the most significant effect. Compared with the OGD group, all drug intervention groups induced a decrease in the apoptotic rate of rBMECs, increased the SOD levels, and decreased the MDA levels (all P<0.01). Compared with the mono-therapy groups, the NXTC+GHI group exhibited a significant improvement in the number of apoptotic rBMECs (P<0.01). All drug intervention groups showed different degrees of increase in membrane potential, and the NXTC+GHI group was higher than the NXTC or GHI group (P<0.01).@*CONCLUSION@#The combinationa application of NXTC and GHI on cerebral I/R injury clearly resulted in protective benefits.

Effect of Competitive Antagonist of Transmembrane Glutamine Flux V-9302 on Apoptosis of Acute Myeloid Leukemia Cell Lines HL-60 and KG-1 / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of V-9302 (an antagonist of transmembrane glutamine flux) on the proliferation and apoptosis of acute myeloid leukemia cells HL-60 and KG-1.@*METHODS@#HL-60 and KG-1 cells at logarithmic phase were treated by different concentrations of V-9302. CCK-8 assay was used to detect the proliferation of the cells. Annexin V-FITC / PI double staining flow cytometry was used to detect the apoptosis of HL-60 and KG-1 cells. The expressions of BAX, BCL-2 and Caspase3 were detected by RT-qPCR and Western blot.@*RESULTS@#V-9302 could significantly inhibit the growth of HL-60 and KG-1 cells. The concentration of V-9302 at 10, 20 μmol/L could significantly promote the apoptosis of HL-60 and KG-1 cells(P<0.05). The results of apoptosis related gene detection showed that when V-9302 was applied to HL-60 and KG-1 cell lines at 10 and 20 μmol/L, the expression levels of Pro-apoptotic protein genes BAX and Caspase3 in HL-60 and KG-1 were significantly higher than those in control group (P<0.05), while the expression level of anti-apoptotic protein gene BCL-2 was significantly lower than that in the control group (P<0.05). The results of Western blot were basically consistent with that of RT-qPCR.@*CONCLUSION@#Competitive antagonist of transmembrane glutamine flux V-9302 can significantly promote the apoptosis of acute myeloid leukemia cell lines HL-60 and KG-1.

Effect of glutamine metabolism on chemoresistance and its mechanism in tumors / 浙江大学学报·医学版

The metabolic reprogramming of tumor cells is characterized by increased uptake of various nutrients including glutamine. Glutamine metabolism provides the required substances for glycolysis and oxidative phosphorylation and affects the homeostasis of carbohydrate，fat and protein metabolism to induce the chemoresistance of tumor cells. Combination of chemotherapeutic agents with inhibitors specific to different components of glutamine metabolic pathway has obtained favorable clinical results on various tumors. Glutamine metabolic pathway plays a role in drug resistance of tumor cells in various ways. Firstly，the dynamic change of glutamine transporters can directly affect intracellular glutamine content thereby causing drug resistance; secondly，tumor stromal cells including adipocyte，fibroblast and metabolite from tumor microenvironment would give rise to immune-mediated drug resistance; thirdly，the expression and activity of key enzymes in glutamine metabolism also has a critical role in drug resistance of tumors. This article reviews the effects of glutamine metabolic pathway in the development of tumor chemoresistance，in terms of transporters，tumor microenvironment and metabolic enzymes，to provide insight for improving the therapeutic efficacy for drug-resistant tumors.

Glutamine use in feeding juvenile pirarucu, Arapaima gigas (Schinz, 1822) / [Uso da glutamina na alimentação de juvenis de pirarucu, Arapaima gigas (Schinz, 1822)]

This study aimed to evaluate glutamine supplementation effects on variables of growth performance, body composition, intestinal morphology and enzymatic aspects of juvenile Arapaima gigas. Research was conducted at the Fish Nutrition and Feeding Laboratory, where 60 examples of pirarucu (initial average weight of 82.12g) were distributed over 15 polyethylene tanks (310L), in a completely randomized design, with five treatments and three repetitions (four fish per experimental unit). Experimental diets were prepared containing five inclusion levels of the amino acid glutamine (0.0, 0.5, 1.0, 1.5 and 2.0%), supplied three times a day for 45 days. Quadratic effect was observed for the variables of growth performance, weight gain, food consumption, food conversion, and specific growth and protein efficiency rates. A significant effect was observed on intestinal villi at the height of the anterior portion and on activity of the enzyme's alkaline proteases, lipase, amylase and aspartate aminotransferase. However, glutamine supplementation had no significant effect on survival rate. Inclusion of 1.02% of glutamine in the diets of juvenile pirarucu improved growth performance and influenced intestinal villi height and activity of important digestive enzymes, favoring nutrient digestion and absorption.(AU)

Objetivou-se avaliar os efeitos da suplementação com glutamina sobre variáveis de desempenho produtivo, composição corporal, morfologia do intestino e aspectos enzimáticos de juvenis de Arapaima gigas. O experimento foi conduzido no Laboratório de Nutrição e Alimentação de Peixes, onde 60 exemplares de pirarucu (peso médio inicial de 82,12g) foram distribuídos em 15 tanques de polietileno (310L), em delineamento inteiramente ao acaso, com cinco tratamentos e três repetições (quatro peixes por unidade experimental). As dietas experimentais foram confeccionadas contendo cinco níveis de inclusão do aminoácido glutamina (0,0; 0,5; 1,0; 1,5 e 2,0%), fornecidas três vezes ao dia, ao longo de 45 dias. Foi observado efeito quadrático para variáveis de desempenho produtivo: ganho de peso, consumo alimentar, conversão alimentar, taxa de crescimento específico e taxa de eficiência proteica. Observou-se ainda efeito significativo sobre a altura das vilosidades da porção anterior do intestino e a atividade das enzimas: proteases alcalinas, lipase, amilase e aspartato aminotransferase. Entretanto, a suplementação com glutamina não influenciou significativamente a sobrevivência dos animais. A adição de 1,02% de glutamina nas dietas para juvenis de pirarucu melhorou o desempenho produtivo e influenciou a altura das vilosidades intestinais e a atividade de enzimas digestivas importantes, favorecendo a digestão e a absorção de nutrientes.(AU)

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

Glutamine protects intestine against ischemia-reperfusion injury by alleviating endoplasmic reticulum stress induced apoptosis in rats

Abstract Purpose Glutamine, as an essential part of enteral nutrition and parenteral nutrition agent, has been widely recognized to be a kind of important intestinal mucosa protectant in clinical practice and experimental research. However, the mechanisms of its protective effects are still not fully understand. Consequently, this study aimed to explore the potential mechanism of glutamine on ischemia-reperfusion (I/R) injury induced endoplasmic reticulum (ER) stress in intestine. Methods An experimental model of intestinal I/R in rats was established by 1 hour occlusion of the superior mesenteric artery followed by 3 hours of reperfusion. Morphologic changes of intestinal mucosa, apoptosis of epithelial cells, and expression of intestinal Grp78, Gadd153, Caspase-12, ATF4, PERK phosphorylation (P-PERK) and elF2αphosphorylation(P-elF2α) were determined. Results After I/R, the apoptotic index of intestinal mucosa epithelial cells observably increased with notable necrosis of intestinal mucosa, and the expressions of Grp78, Gadd153, Caspase-12, ATF4, P-PERK and P-elF2αall were increased. However, treatment with glutamine could significantly relieve intestinal I/R injury and apoptosis index. Moreover, glutamine could clearly up-regulate the expression of Grp78, restrain P-PERK and P-elF2α, and reduce ATF4, Gadd153 and Caspase-12 expressions. Conclusion Glutamine may be involved in alleviating ER stress induced intestinal mucosa cells apoptosis.

Suplementação para controle de diarreia em idosos hospitalizados com nutrição enteral / Supplementation for diarrhea control in hospitalized geriatric patients on enteral nutrition

OBJETIVO: Comparar resultados da suplementação com prebiótico, probiótico e simbiótico para o controle da diarreia em pacientes idosos recebendo terapia nutricional enteral durante o internamento em um hospital escola de Curitiba, Paraná. MÉTODOS: O estudo foi retrospectivo, por análise de prontuários correspondentes aos atendimentos realizados entre 2014 e 2018. RESULTADOS: Obteve-se um total de 75 pacientes. O tempo de ocorrência de diarreia variou de 1 a 16 dias, sendo a média de 2,69 dias após a instituição de terapêutica para restabelecimento da microbiota intestinal. Quanto às terapias instituídas, foram encontradas oito possíveis prescrições de suplementos isolados e/ou combinados, como primeira escolha. Dos pacientes analisados, 52% trocaram de suplementação ao longo da ocorrência da diarreia; alguns chegando a utilizar até cinco diferentes produtos. Dos 48% de pacientes que utilizaram um único produto/combinação do início ao fim da diarreia, de modo geral iniciaram com uma dose maior e foram diminuindo ao longo do tempo, sendo que os que começaram com uma dose menor tiveram que aumentá-la para interromper a diarreia. Além disso, houve significância estatística quando comparado o tempo de diarreia entre pacientes que receberam um único produto/combinação e os que fizeram trocas de suplemento ao longo do tratamento. CONCLUSÃO: Estabelecer uma prescrição única, seja de produtos isolados ou combinados, e permanecer com ela, além de iniciar com uma dose maior, parece mais efetivo no controle da diarreia em idosos hospitalizados, reforçando a importância de se estabelecer um protocolo para prescrição.

OBJECTIVE: To compare results of prebiotic, probiotic and synbiotic supplementation for the control of diarrhea in older patients receiving enteral nutritional therapy during hospitalization at a school hospital in Curitiba, state of Paraná. METHODS: The study was retrospective, by analysis of medical records corresponding to the visits performed between 2014 and 2018. RESULTS: A total of 75 patients were analyzed. The time of occurrence of diarrhea ranged from 1 to 16 days, with a mean of 2.69 days after the onset of therapy for reestablishment of the intestinal microbiota. As for the therapies introduced, 8 possible prescriptions of isolated and / or combined supplements were found as the first choice. Of the patients analyzed, 52% switched from supplementation during the occurrence of diarrhea; some using up to 5 different products. Of the 48% of patients who used a single product / combination from the beginning to the end of diarrhea, they generally started with a higher dose and decreased over time, with those starting at a lower dose having to increase it to stop diarrhea. In addition, there was statistical significance when comparing the time of diarrhea between patients who received a single product / combination and those who did supplemental exchanges throughout the treatment. CONCLUSION: Establishing a single prescription, whether of isolated or combined products and sticking to it, besides starting with a higher dose, seems more effective in controlling diarrhea in hospitalized geriatric patients, reinforcing the importance of establishing a protocol for prescription.

Anxiolytic properties of compounds that counteract oxidative stress, neuroinflammation, and glutamatergic dysfunction: a review

Objective: Anxiety disorders are highly prevalent and the efficacy of the available anxiolytic drugs is less than desired. Adverse effects also compromise patient quality of life and adherence to treatment. Accumulating evidence shows that the pathophysiology of anxiety and related disorders is multifactorial, involving oxidative stress, neuroinflammation, and glutamatergic dysfunction. The aim of this review was to evaluate data from animal studies and clinical trials showing the anxiolytic effects of agents whose mechanisms of action target these multiple domains. Methods: The PubMed database was searched for multitarget agents that had been evaluated in animal models of anxiety, as well as randomized double-blind placebo-controlled clinical trials of anxiety and/or anxiety related disorders. Results: The main multitarget agents that have shown consistent anxiolytic effects in various animal models of anxiety, as well in clinical trials, are agomelatine, N-acetylcysteine (NAC), and omega-3 fatty acids. Data from clinical trials are preliminary at best, but reveal good safety profiles and tolerance to adverse effects. Conclusion: Agomelatine, NAC and omega-3 fatty acids show beneficial effects in clinical conditions where mainstream treatments are ineffective. These three multitarget agents are considered promising candidates for innovative, effective, and better-tolerated anxiolytics.

Glutamine Supplementation Ameliorates Chronic Stress-induced Reductions in Glutamate and Glutamine Transporters in the Mouse Prefrontal Cortex

Chronic immobilization stress (CIS) induces low levels of glutamate (Glu) and glutamine (Gln) and hypoactive glutamatergic signaling in the mouse prefrontal cortex (PFC), which is closely related to the Glu-Gln cycle. A Gln-supplemented diet ameliorates CIS-induced deleterious changes. Here, we investigated the effects of CIS and Gln supplementation on Glu-Gln cycle-related proteins to characterize the underlying mechanisms. Using the CIS-induced depression mouse model, we examined the expression of 11 proteins involved in the Glu-Gln cycle in the PFC. CIS decreased levels of glutamate transporter 1 (GLT1) and sodium-coupled neutral amino acid transporter (SNAT) 1, SANT2, SNAT3, and SNAT5. Gln supplementation did not affect the non-stressed group but significantly increased GLT1 and SNATs of the stressed group. By immunohistochemical analysis, we confirmed that SNAT1 and SNAT2 were decreased in neurons and GLT1, SNAT3, and SNAT5 were decreased in astrocytes in the medial PFC of the stressed group, but Gln-supplemented diet ameliorated these decrements. Collectively, these results suggest that CIS may cause depressive-like behaviors by decreasing Glu and Gln transportation in the PFC and that a Gln-supplemented diet could prevent the deleterious effects of CIS.

Efficacy of parenteral glutamine supplementation in adult hematopoietic stem cell transplantation patients

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) patients need parenteral nutrition because of nausea, vomiting, and mucositis caused by conditioning regimens. The demand for glutamine increases during the HSCT period. We evaluated the effects of glutamine-containing parenteral nutrition on the clinical outcomes of HSCT patients. METHODS: In this retrospective analysis, we reviewed HSCT patients from Seoul National University from August 2013 to July 2017. Depending on their glutamine supplementation status, 91 patients were divided into 2 groups: glutamine group (N=44) and non-glutamine group (N=47). We analyzed the rate of weight change, infection (clinically/microbiologically documented), complications (duration of mucositis and neutropenia, acute graft versus host disease), and 100-days mortality in each group. RESULTS: Regarding the clinical characteristics of the patients, there were no significant differences between the 2 groups except that there was a larger proportion of myeloablative conditioning regimen in the glutamine group (P=0.005). In the glutamine group, the average number of days of glutamine use, parenteral nutrition, and mucositis was 7.6±1.4, 14.6±9.9, and 13.3±9.5, respectively. Furthermore, multivariate analysis revealed odds ratios of 0.37 (95% CI, 0.14–0.96; P=0.042) and 0.08 (95% CI, 0.01–0.98; P=0.048) for clinically documented infection and 100-days mortality, respectively, in the glutamine group. CONCLUSION: Results showed that the glutamine group had less clinically documented infection and 100-days mortality than the non-glutamine group, but the other outcomes did not show significant differences. The extended duration of glutamine supplementation according to the period of total parenteral nutrition and mucositis should be considered.