Glutamine dipeptide supplementation improves clinical responses in patients with diabetic foot syndrome

ABSTRACT The effect of glutamine dipeptide (GDP) supplementation in patients with diabetic foot syndrome was evaluated. A total of 22 patients took part in the study. GDP was supplied in 10 g sachets, and was dissolved in water immediately before use, with ingestion once a day, after lunch or after dinner (20 g/day) over a period of 30 days. Quantification of foot insensitive areas, oxidative stress, blood cytokines, and biochemical, hematological and toxicological parameters was performed before and after GDP supplementation. We observed an increase in blood levels of interferon-α (P=0.023), interferon-γ (P=0.038), interleukin-4 (P=0.003), interleukin-6 (P=0.0025), interleukin-7 (P=0.028), interleukin-12 p40 (P=0.017), interleukin-13 (P=0.001), leukocytes (P=0.037), eosinophils (P=0.049), and typical lymphocytes (P<0.001) due to GDP administration. In addition, we observed a reduced number (P=0.048) of insensitive areas on the foot, and reduction (P=0.047) of fasting hyperglycemia. Patients also showed increased blood high density lipoprotein (P<0.01) and protein thiol groups (P=0.004). These favorable results were associated with the absence of renal and hepatic toxicity. These results are of clinical relevance, since supplementation with GDP over 30 days improved clinical responses in patients with diabetic foot syndrome.

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

Evaluation of glutamine, ammonia and cortisol in endurance horses during races of different lengths / Avaliação da glutamina, amônia e cortisol em cavalos de enduro durante corridas de diferentes distâncias

Glutamine is the most abundant free amino acid in the bodies of mammals. It is very important for cell proliferation, metabolism of ammonia and energy production in skeletal muscles, especially during aerobic exercise. The aim of this study was to investigate if serum levels of glutamine and ammonia could be models for studying the dynamics of this important amino acid during exercise in endurance horses, as these animals are subjected to prolonged and intense aerobic workloads. Thirty-three endurance horses that competed in the categories of 80 km (n = 13), 120 km (n = 14) and 160 km (n = 5) in four races were tested. There was no statistical difference in the levels of glutamine between moments (P > 0.05), but there was a significant increase in plasma levels of ammonia right after the race (P 0.05). This increase is more marked in the horses that ran 80 km, followed by those that ran 120 km and finally by those running 160 km. There was a positive correlation between the levels of ammonia and glutamine in categories 160, 120 and 80 km (r = 0.209, r = 0.151 and r = 0.176, respectively; P 0.05). Oscillations in serum levels of glutamine are difficult to detect in endurance horses, since oxidative muscle fibers are important reservoirs of this amino acid. Also, it is difficult to know about glutamine consumption because of the organic reserves and ease of metabolization; hence, measuring ammonia as an indicator of consumption of glutamine. We concluded that ammonia and cortisol levels can serve as indirect measures of the use of glutamine during aerobic exercise and both can be markers of effort.

A glutamina e o aminoácido livre mais abundante no organismo de mamíferos, muito importante para a proliferação celular, metabolismo da amônia e para produção de energia na musculatura esquelética, principalmente durante o exercício aeróbico. O objetivo do presente trabalho foi investigar se os níveis séricos de glutamina, amônia e cortisol, em cavalos de enduro, podem ser modelos de estudo da dinâmica deste importante aminoácido durante o exercício, pois estes animais são submetidos a esforço aeróbico prolongado e intenso. Foram utilizados 33 cavalos de enduro que competiram nas categorias de 80 km (n = 13), 120 km (n = 14) e 160 km (n = 5) em quatro provas. Não houve diferença estatística nos níveis de glutamina entre os momentos de coleta (P > 0,05). Houve aumento significativo dos níveis plasmáticos de amônia apos o termino das provas (P 0,05). Esse aumento foi mais marcado nos cavalos de 80 km, seguido pelos de 120 e por ultimo os de 160 km. Houve correlação positiva entre os níveis de amônia e glutamina nas categorias de 160, 120 e 80 km (r = 0,209, r = 0,151 e r = 0,176, respectivamente; P 0,05). Oscilações séricas dos níveis de glutamina são difíceis de se detectar em cavalos de enduro, já que as fibras musculares oxidativas são importantes reservatórios desse aminoácido. Ainda, e difícil saber sobre consumo de glutamina dadas as reservas orgânicas e a facilidade de metabolização. A concentração sérica de amônia pode servir como indicador indireto do consumo de glutamina. Assim, concluiu-se que as dosagens de amônia e cortisol podem servir como medidas indiretas da utilização da glutamina durante o exercício aeróbico e ambas como marcadores de esforço. (AU) 

Effects of glucose and glutamine concentrations in human dental pulp stem cells viability. An approach for cell transplantation

PURPOSE: To evaluate microscopic behavior and viability of dental pulp stem cells under glucose and glutamine deprivation. METHODS: Human tooth tissues were minced in isolated pieces and cultured until the desired cellular proliferation for experimental phases. Cells were cultured under variations of glucose and glutamine in both serum presence and absence, and then those cells were evaluated according to number and viability by MTT assay. The confocal microscopy analyzed cytoskeleton, nucleus, and mitochondria integrity. RESULTS: A low concentration of glucose favored cellular viability and microscopic behavior; the presence of glutamine in culture medium was favorable only when associated with glucose. The cellular biological potential in culture could be preserved in serum absence if nutritional requirements are adequate. CONCLUSION: Cell microscopic behavior and viability have demonstrated better patterns on serum-free low glucose culture medium with glutamine deprivation. .

Efeito de diferentes protocolos de treinamento de força sobre parâmetros morfofuncionais, hormonais e imunológicos / Effects of different resistance training protocols over the morphofunctional, hormonal and immunological parameters

O objetivo do estudo foi examinar a influência de dois diferentes protocolos de treinamento de força sobre parâmetros antropométricos (peso, IMC, massa gorda), funcionais (teste de 1-RM e teste de repetições máximas) e relacionados ao sistema endócrino (concentração de testosterona e de cortisol) e ao sistema imunológico (concentração de glutamina e de IgG). Participaram do estudo 12 homens treinados (27,4 ± 4,8 anos). Esses indivíduos foram aleatoriamente divididos em dois grupos, que posteriormente foram submetidos a dois protocolos de treinamento distintos: Múltiplas séries (MS) e Tri-set (TS). Amostras de sangue foram coletadas antes e depois de uma sessão de exercício de força, no início e no final do período de oito semanas de treinamento. Não foram observadas alterações nos parâmetros morfo-funcionais (com exceção do teste de repetições máximas para o agachamento). Com relação aos parâmetros endócrinos, foi observado que o TS provocou aumento significativo do cortisol, imediatamente após a sessão de treino, tanto no início como no final das oito semanas (p < 0,05). Ao observar o comportamento da relação testosterona para cortisol (T:C), pode-se notar um marcante aumento no grupo submetido ao protocolo MS após oito semanas de treinamento (p < 0,05). Com relação aos parâmetros imunológicos, não foi observada alteração na concentração de imunoglobulina G. A concentração de glutamina sofreu decréscimo após oito semanas em ambos os grupos. Esse decréscimo foi mais acentuado no grupo TS (p < 0,05). Os resultados obtidos sugerem que o método TS impôs maior estresse ao organismo. Além disso, os dados também indicam que o protocolo MS promove um ambiente mais propício ao anabolismo, após oito semanas de treinamento. Entretanto, ambos os métodos falharam em promover alterações significativas nos parâmetros morfofuncionais.

The purpose of this study was to assess the influence of two different resistance training protocols on the anthropometric (weight, BMI, fat mass), functional parameters (1-MR test, and maximal repetition test) and the parameters related to the endocrine system (testosterone and cortisol concentrations), as well as to the immunological system (glutamine and IgC concentrations). The study was composed by twelve trained men (27.4 ± 4.8 years), who were randomly divided in two groups that later were submitted to two different training protocols: the Multiple Series (MS), and Tri-set (TS). Blood samplings were collected before and after an resistance training session in the beginning and the end of the 8 weeks training period. It was observed no alterations in the morphofunctional parameters (except as to the maximal repetition test for the squat). As to the endocrine parameters, it was observed that the TS caused a significant increase in the cortisol immediately after the training session both in the beginning and in the end of the eight weeks (p < 0.05) period. Upon the observation of the testosterone vs. cortisol ratio (T:C) behavior, it can be observed a noticeable increase in the group submitted to the MS protocol after the 8 weeks training period (p < 0.05). As to the immunological parameters, it was observed no alterations in the concentration of the immunoglobulin G. The concentration of the glutamine suffered a decrease after 8 weeks in both groups. That decrease had a higher accentuation in the TS group (p < 0.05). Results attained suggest that the TS method imposed a higher stress to the body. Furthermore, these data also indicate that the MS protocol promotes a more propitious environment to the anabolism after the 8 weeks training period. However, both methods did not succeed in promoting significant changes in the morphofunctional parameters.

El objetivo del estudio fué examinar la influencia de 2 protocolos diferentes de entrenamiento de fuerza sobre parámetros antropométricos (peso, IMC, masa grasa), funcionales (prueba de 1-RM y prueba de repeticiones máximas) y los parámetros relacionados al sistema endocrino (concentraciones de testosterona y de cortisol) y al sistema inmunológico (la concentración del glutamina y de IgG). Participaron en el estudio 12 hombres entrenados (27,4 ± 4,8 años). Estos individuos fueron aleatoriamente divididos en 2 grupos que después se sometieron a 2 protocolos de entrenamiento en dos series diferentes, Múltiple (MS) y Tri-juego (TS). Las muestras de sangre eran reunidos antes y después de una sesión de ejercicio de fuerza, al principio y en el fin del período de 8 semanas de entrenar. No se observaron alteraciones en los parámetros morfo-funcionales (salvo la prueba de repeticiones máximas para el agachamiento). Con respecto a los parámetros endócrinos, se observó que TS provocó un aumento significante del cortisol, inmediatamente después de la sesión de entrenamiento, al principio y en el fin de las 8 semanas (p < 0,05). Al observar la conducta de la testosterona en relación con el cortisol (T:C), puede notarse un aumento excelente en el grupo sometido al MS protocolar después de 8 semanas de entrenar (p < 0,05). Con respecto a los parámetros inmunológicos, no se observó alteración en la concentración de la imunoglobulina G; la concentración del glutamina sufrió una disminución después de 8 semanas en ambos grupos. Esta disminución se acentuó más en el TS de grupo (p < 0,05). Los resultados obtenidos sugieren que el método TS impuso una tensión más grande al organismo. Además, los datos también indican que el MS protocolar promueve una atmósfera más favorable al anabolismo, después de 8 semanas de entrenar. Sin embargo, ambos métodos fallaron promoviendo las alteraciones significantes en los parámetros morfo-funcionales.

Current concepts of the relationship between cholesterol fed rats and endogenous amino acids

The effect of long term feeding of cholesterol 100 mg/kg/day for 20 weeks on free amino acid patterns in plasma and liver contents in rats was investigated using LKB-amino acid analyzer [Biochrom Ltd., Cambridge, England] and correlated these results with their effects on plasma levels of cholesterol, triglyceride, lipoprotein and Gamma Glutamyl Transferase [yGT]. indicate that all amino acid contents in the liver were significantly decreased in the cholesterol feed animal group. The amplitude of reductions varied between 40-100%. Glutamine and tryptophan were not detected in the liver of cholesterol feed group. The plasma concentrations of taurine, glutamate, alanine, valine and phenylalanine were elevated whose mean percentage increases were 48 +/- 4, 40 +/- 3, 25 +/- 3 and 35 +/- 4 respectively. These increases were associated with significant decrease in the concentration of ornithine [37%]. Meanwhile proline was not detected in the plasma of treated animals. Also, plasma cholesterol, triglyceride, lipoproteins and yGT were determined by colorimetric methods using Kits from Boehringer Mannheim [GmbH]. Results indicated that feeding cholesterol significantly increased the plasma yGT activity. In these experimental conditions the chronic intake of cholesterol had no significant effects on plasma cholesterol or other plasma lipids parameters tested except plasma triglyceride which was significantly increased, these results indicated that there are interactions between cholesterol intake and hepatic glutamine and tryptophan as well as plasma proline and these interaction mechanisms may be considered the factor generating metabolic events, which play physiological functions in the regulation of plasma cholesterol. Thus, under pathological conditions there is an imbalance between these interaction mechanisms which cause an increase in the circulating levels of cholesterol, leading to pathological processes such as hyperlipidemias, atherosclerosis and bile stones

Free glutamic acid content of milk in Indian mothers.

Free amino acids levels in the milk of 11 healthy Indian mothers were determined using automatic precolumn derivatization procedure. The aim of the study was to find out the relative concentration of glutamic acid and glutamine in the milk. Glutamic acid is the dominant free amino acid found in the milk of Indian mothers. Glutamic acid and glutamine together formed the major nonessential amino acids present in the human milk. Although glutamic acid has been shown to be the major amino acid in human milk in many studies, to the best our knowledge, this is the first report to confirm that glutamic acid is the most abundant amino acid in milk in Indian mothers.

Diagnostic value of adenosine deaminase and glutamine in tuberculous pleural effusions

Two biochemiccal markers related to lymphocyte functions are adenosine deaminase [ADA] enzyme and glutamine [Glu.]. The discriminating value of both of these indices in differentiating tuberculous from other types of pleural effusions was studied. The present work included 23 patients with tuberculous pleural effusions, 14 patients with malignant effusions as well as 5 patients with post-pneumonic effusions. Pleural fluids and sera were obtained from each participant and used for determination of pleural and serum ADA activity as well as pleural glutamine. It was found that tuberculous pleural effusions had signficantly higher ADA than other types of effusions, while malignant effusions had significantly higher glutamine levels. When ADA activity of more than 30.2 U/L is considered, diagnostic tests of tuberculous effusions showed a sensitivity of 100%, specificity of 79%. A pleural fluid, serum ADA ratio above 2.1 was found in 100% of tuberculous and in 7% of carcinomatous effusions [sensitivity 100%, specificity 93%, diagnostic accuracy 97%]. All postpneumonic effusions had a ratio below 2.1 A pleural fluid glutamine level above 2.2 mg/dl, selected as the discriminating level, was found in 100% of malignant, 4% tuberculous and 20% of postpneumonic effusions. The combined assay of pleural ADA pleural/serum ADA and pleural glutamine provide in-formation in the deferential diagnosis of pleural effusions, especially differentiating tuberculous from malignant ones. These are easy, cheap and could be performed in any hospital lab, as a routine to help diagnosis of pleural effusions

Esquizofrenia paranoide: aminoácidos libres en el líquido cefalorraquídeo / Paranoid schizophrenia: free amino acids in cerebrospina

En el presente estudio los autores reportan un incremento de la concentración de glutamina en el líquido cefalorraquídeo de siete pacientes esquizofrénicos paranoides, sin cambios en los niveles de los 20 aminoácidos libres restantes

Transamination and glutamate deamination in Rana hexadactyla during induced ammonia toxicity.

Glutamate dehydrogenase (GDH) and the transaminases namely aspartate aminotransferase (AAT) and alanine aminotransferase (AIAT) were estimated in the muscle, liver, kidney, and brain of control and ammonium acetate administered frogs. The results indicated tissue specific responses during induced ammonotoxemia. The inherent endogenous ammonia production decreased in all the tissues. 2-Keto glutarate production appears to be the other main adaptive feature as a result of slightly stepped up transdeamination patterns.