Correlation between intestinal and respiratory flora and their metabolites in a rat pneumoconiosis model / 中华劳动卫生职业病杂志

Objective: Differential flora and differential metabolites shared by the intestinal and respiratory tracts of rats were screened to analyze the possible role of changes in intestinal flora and metabolites in the progression of pneumoconiosis in rats. Methods: In April 2020, 18 SD rats were randomly divided into three groups (control group, coal mine dust group and silica group, 6 in each group) , rats in the coal mine dust group and silica group were perfused with 1 ml of 50 mg/ml coal mine well dust suspension and silica suspension by nontracheal exposure, respectively. While rats in the control group were perfused with an equal dose of sterilized normal saline. Twenty four weeks after dust staining, rat feces, throat swabs, and lung lavages were collected. 16SrDNA gene sequencing and UHPLC-QTOF-MS untargeted metabolomics were used to analyze the flora and metabolites in feces, throat swabs and lung lavage fluid of rats in each group, to screen for shared differential flora and shared differential metabolites in intestinal and respiratory tract, and the correlation analysis between the differential flora and metabolites was performed using Spearman's statistics. Results: Compared with the control group, a total of 9 species shared differential flora between intestinal and respiratory tract were screened at phylum level, and a total of 9 species shared differential genus between intestinal and respiratory tract were screened at genus level in the coal mine dust group, mainly Firmicutes, Actinobacteria, Streptococcus, Lactobacillus, etc. Compared with the control group, a total of 9 shared differential flora were screened at the phylum level, and a total of 5 shared differential genus were screened at the genus level in the silica group, mainly Proteobacteria, Actinobacteria, Allobactera, Mucilaginibacter, etc. Compared with the control group, a total of 7 shared differential metabolites were screened for up-regulation of Stigmatellin, Linalool oxide and Isoleucine-leucine in both intestinal and respiratory tract in the coal mine dust group. Compared with the control group , a total of 19 shared differential metabolites werescreened in the silica group, of which Diethanolamine, 1-Aminocyclopropanecarboxylic acid, Isoleucine-leucine, Sphingosine, Palmitic acid, D-sphinganine, 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine, and 1-Stearoyl-2-oleoyl-sn-glycerol 3-phosphocholine were up-regulated in both the intestinal and respiratory tract. Conclusion: There is a translocation of intestinal and respiratory flora in pneumoconiosis rats, and rats have an imbalance of lipid metabolism during the progression of pneumoconiosis.

Branched-Chain amino acids intake is negatively related to body adiposity in individuals at cardiometabolic risk / Consumo de aminoácidos de cadeia ramificada está negativamente relacionado com adiposidade corporal em indivíduos com risco cardiometabólico

ABSTRACT Objective To assess the relationship between branched-chain amino acids intake in the current diet and the metabolic and body adiposity markers in a population at cardiovascular risk. Methods This is a cross-sectional study with 282 adults and elderly people from the Cardiovascular Health Care Program of the Universidade Federal de Viçosa. Sociodemographic, anthropometric and body composition data, as well as metabolic biomarkers were collected using standardized protocols. Dietary intake of branched amino acids was assessed using a 24-hour recall. Results Individuals with a higher branched-chain amino acids intake (≥2.6g/day, median value) had lower body fat (29.6 vs 32.2%; p=0.019), and higher serum ferritin (113.2 vs. 60.1mg/dL; p=0.006) and uric acid concentrations (4.4 vs. 4.0; p=0.023). In addition, a lower prevalence of overweight and excessive abdominal fat (p<0.05) was found in the individuals with higher branched-chain amino acids intake. They also had a higher daily intake of fiber, copper, zinc, magnesium, and iron, as well as a lower intake of total lipids. Conclusion In the present study, the intake of branched amino acids is negatively related to total and central adiposity, but more studies are needed to fully elucidate this possible relationship. (Brazilian Registry of Clinical Trials, code RBR-5n4y2g).

RESUMO Objetivo Avaliar a relação entre o consumo de aminoácidos de cadeia ramificada na dieta atual e os marcadores de adiposidade metabólica e corporal em uma população com perfil de elevado risco cardiovascular. Métodos Trata-se de um estudo transversal com 282 adultos e idosos do Programa de Atenção Cardiovascular da Universidade Federal de Viçosa. Dados sociodemográficos, antropométricos e de composição corporal, além de biomarcadores metabólicos, foram coletados utilizando protocolos padronizados. O consumo alimentar de aminoácidos ramificados foi avaliado através de um recordatório de 24 horas. Resultados Indivíduos com maior consumo de aminoácidos de cadeia ramificada (≥2,6g/dia, valor da mediana) apresentaram menores valores de gordura corporal (29,6 vs 32,2%; p=0,019) e maiores valores de séricos de ferritina (113,2 vs. 60,1mg/dL; p=0,006) e ácido úrico (4,4 vs. 4,0; p=0,023). Além disso, foi encontrada uma menor prevalência de sobrepeso e excesso de gordura abdominal (p<0,05) nos indivíduos com maior consumo de aminoácidos de cadeia ramificada. Eles também apresentaram um maior consumo diário de fibra, cobre, zinco, magnésio e ferro, além de um menor consumo de lipídios totais. Conclusão No presente estudo, o consumo de aminoácidos ramificados está negativamente relacionado à adiposidade total e central, porém mais estudos são necessários para elucidar completamente essa possível relação. (Registro Brasileiro de Ensaios Clínicos, código RBR-5n4y2g)

A 1-year-old female with maple syrup urine disease presenting with acrodermatitis-enteropathica-like lesions

@#<p>A 1-year-old female with maple syrup urine disease presenting with erythematous, partially eroded plaques on the trunk, anogenital area, and extremities experienced metabolic crisis. The skin lesions appeared at 11 months of age and was thought to result from amino acid imbalance secondary to erratic supplementation of specialized milk formula devoid of isoleucine, leucine, and valine. Serial urine monitoring showed persistent ketones and elevated serum leucine and valine. The patient was managed with emollients, intralipid 20%, and addition of isoleucine and valine supplements to counter the neurotoxic effect of leucine. After 8 days of proper feeding and continuous emollient application, the lesions improved and skin biopsy revealed superficial perivascular dermatitis. Although a decrease in erythema and desquamation was noted, the patient had persistent cerebral edema and continued to deteriorate.</p>

Exigência de valina e isoleucina para codornas de corte no período de um a 14 dias e de 15 a 35 dias de idade / Valine and isoleucine requirement for meat quails in the period of 1 to 14 days and of 15 to 35 days old

Foram conduzidos dois experimentos, com o objetivo de estimar a exigência de valina e isoleucina digestível para codornas de corte em crescimento. No experimento 1, foram utilizadas 2.160 codornas de corte de um dia de idade, não sexadas, distribuídas em um delineamento inteiramente ao acaso, com arranjo fatorial 4x4, sendo quatro níveis de valina digestível (0,90, 1,10, 1,30 e 1,50%) e quatro níveis de isoleucina digestível (0,80, 1,00, 1,20 e 1,40%), totalizando 16 tratamentos, com três repetições e 45 codornas por unidade experimental. O peso corporal, o ganho de peso e a biomassa corporal acumulada aumentaram linearmente em função dos níveis de valina digestível e foram influenciados de forma quadrática, estimando 1,38% de isoleucina digestível. No experimento 2, foram utilizadas 1.440 codornas de corte não sexadas, distribuídas em um delineamento inteiramente ao acaso, com arranjo fatorial 4x4, sendo quatro níveis de valina digestível (0,82, 1,02, 1,22 e 1,42%) e quatro níveis de isoleucina digestível (0,73, 0,93, 1,13 e 1,33%), totalizando 16 tratamentos, com três repetições e 30 codornas por unidade experimental. O desempenho de codornas de corte, no período de 15 a 35 dias de idade, não foi influenciado pelo aumento dos níveis de valina e isoleucina digestível nas rações experimentais. Conclui-se que as exigências de valina e isoleucina digestível para o máximo desempenho de codornas de corte, no período de um a 14 dias de idade, foram de 1,50% e 1,38%, respectivamente, correspondendo às relações valina:lisina de 95% e isoleucina:lisina de 88%. Os menores níveis avaliados, no período de 15 a 35 dias de idade, de 0,82% de valina digestível e de 0,73% de isoleucina digestível, correspondendo às relações valina:lisina de 52% e isoleucina: lisina de 46%, foram suficientes para satisfazer as exigências nutricionais das codornas de corte sem comprometer o desempenho.(AU)

Two experiments were carried out in order to estimate the requirements of digestible valine and isoleucine for growing meat quails. In experiment 1, 2160 meat quails with 1 day of age, not sexed, were used, distributed in a completely randomized design with a 4x4 factorial arrangement, with four levels of digestible valine (0.90, 1.10, 1.30, and 1.50%) and four levels of digestible isoleucine (0.80, 1.00, 1.20, and 1.40 %), totaling 16 treatments with three replications and 45 quails per experimental unit. The body weight, the weight gain, and the accumulated body biomass linearly increased in function of digestible valine levels and were influenced in a quadratic way estimating 1.38% of digestible isoleucine. In experiment 2, 1440 meat quails, not sexed, were used, distributed in a completely randomized design with a 4x4 factorial arrangement, with four levels of digestible valine (0.82, 1.02, 1.22, and 1.42%) and four levels of digestible isoleucine (0.73, 0.93, 1.13, and 1.33%), totaling 16 treatments with three replications and 30 quails per experimental unit. The performance of meat quails from 15 to 35 days of age was not affected by increased levels of digestible valine and digestible isoleucine in the experimental diets. It was concluded that the requirements of digestible valine and digestible isoleucine for maximum performance of meat quail from one to 14 days of age were 1.50% and 1.38%, respectively, corresponding to relations: valine: lysine of 95% and isoleucine: lysine of 88%. The lower levels evaluated, in the period of 15 to 35 days old, 0.82% digestible valine and 0.73% of digestible isoleucine, corresponding to relations valine: lysine of 52% and isoleucine: lysine of 46% were sufficient to meet the nutritional requirements of meat quails without compromising performance.(AU)

Plasma amino acid profiling of "same symptom for different disease" in Uyghur medicine based on high-performance liquid chromatography / 中南大学学报(医学版)

OBJECTIVE@#To determine the plasma amino acid metabolism of "same symptom for different diseases" in different cancer patients in Uyghur medicine.@*METHODS@#Plasma amino acid concentration was tested by high-performance liquid chromatography (HPLC) in cancer patients with different symptom, and the spectral profiles were subjected to a t-test for statistical significance.@*RESULTS@#Compared with the healthy group, lung cancer, cervical cancer, breast cancer and gastric cancer patients with abnormal Savda had lower concentration of plasma amino acids except some amino acids. Lung cancer patients with abnormal Savda had higher concentration of plasma phenylalanine, serine, cystine, valine, isoleucine, leucine and aspartic acid than Unsavda patients (P<0.05). Cervical cancer patients with abnormal Savda had low concentration of plasma arginine, but higher concentration of plasma cystine than Unsavda patients (P<0.05). Breast cancer patients with abnormal Savda had higher concentration of plasma leucine, serine, taurine, cystine, tyrosine, valine, isoleucine and asparagine than Unsavda patients (P<0.05). Gastric cancer patients with abnormal Savda had high concentration of plasma cystine but lower concentration of plasma phenylalanine, threonine and arginine than Unsavda patients (P<0.05).@*CONCLUSION@#Different tumor patients with abnormal Savda have common characteristics and significant differences.

Identification of Two Novel BCKDHB Mutations in Korean Siblings with Maple Syrup Urine Disease Showing Mild Clinical Presentation

Maple syrup urine disease (MSUD) is a disorder that involves the metabolism of branched chain amino acids, arising from a defect in branched-chain alpha-keto acid dehydrogenase complex. Mutations have been identified in the BCKDHA, BCKDHB, or DBT genes, which encode different subunits of the BCKDH complex. Although encephalopathy and progressive neurodegeneration are its major manifestations, the severity of the disease may range from the severe classic type to milder intermediate variants. We report two Korean siblings with the milder intermediate MSUD who were diagnosed with MSUD by a combination of newborn screening tests using tandem mass spectrometry and family genetic screening for MSUD. At diagnosis, the patients' plasma levels were elevated for leucine, isoleucine, valine, and alloisoleucine, and branched-chain alpha-keto acids and branched-chain alpha-hydroxy acids were detected in their urine. BCKDHA, BCKDHB, and DBT analysis was performed, and two novel mutations were identified in BCKDHB. Our patients were thought to have the milder intermediate variant of MSUD, rather than the classic form. Although MSUD is a typical metabolic disease with poor prognosis, better outcomes can be expected if early diagnosis and prompt management are provided, particularly for milder forms of the disease.

Resistance to thyroid hormone due to a novel mutation of thyroid hormone receptor beta gene

Resistance to thyroid hormone (RTH) is a rare inherited syndrome characterized by diminished response of the target tissue to thyroid hormone caused, in the majority of cases, by mutation of the thyroid hormone receptor beta (THRbeta) gene. Despite elevated serum levels of free thyroid hormones and thyroid stimulating hormone (TSH), the paucity of symptoms and signs of thyroid dysfunction suggest RTH. We report the case of a 9-year-old girl with goiter. Her thyroid function tests showed increased serum levels of free thyroxine, triiodothyronine, and TSH. The genetic analysis of THRbeta confirmed a novel mutation in exon 9; this was a heterozygous C-to-T change in the 327th codon, substituting threonine for isoleucine (T327I).

Annual Report on External Quality Assessment of Biochemical Genetics in Korea (2013) / 임상검사와정도관리

Two trials of external quality assessment (EQA) of conventional newborn screening tests for phenylketonuria, galactosaemia, congenital adrenal hyperplasia, maple syrup urine disease, homocystinuria, and congenital hypothyroidism, as well as of newborn screening tests using tandem mass spectrometry were performed in 2013. A total of 32 specimens in the form of dried blood spots were distributed to 16 laboratories and the response rate of these laboratories was 100%. Total T4, free T4, 17-hydroxyprogesterone, leucine, isoleucine, galactose, methionine, alanine, C8/C2, C8/C10, and C5-OH did not meet the accepted performance criteria. The mean, standard deviation, coefficient of variation, median, and cut-offs were evaluated for each analyte in the newborn screening tests. Two trials of EQA for the analyses of methylmalonic acid, vanillylmandelic acid, very long fatty acids, organic acids, and amino acids were also performed. A well-designed EQA program and continuous education would improve the performance of biochemical genetic tests.

Effect of Trigonella foenum-graecum 4-hydroxyisoleucine on high-glucose induced insulin resistance in 3T3-L1 adipocytes of mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of 4-hydroxyisoleucine (4-HIL), an active component of Trigonella Foenum-graecum L. on high glucose induced insulin resistance (IR) in 3T3-L1 adipocytes, and to explore underlying molecular mechanisms.</p><p><b>METHODS</b>3T3-L1 adipocytes were treated with 25 mmol/L glucose and 0.6 nmol/L insulin to induce IR. They were intervened by different concentrations of 4-HIL (at 5, 10, and 20 micromol/L). [3H]-Deoxy-D-glucose up-taking method was used to detect the glucose uptake. The mRNA expression of cellular tumor necrosis factor-alpha (TNF-alpha) was detected by polymerase chain reaction (PCR). The content of TNF-alpha in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Palmitic acid (PA) acted as the control.</p><p><b>RESULTS</b>After intervened by 25 mmol/L glucose and 0.6 nmol/L insulin for 18 h, the insulin-stimulated glucose transportation in 3T3-L1 adipocytes was inhibited by 63%. The mRNA expression of cellular TNF-alpha in adipocytes significantly increased, when compared with that in normal adipocytes (P < 0.05). The level of TNF-alpha secreted in the culture supernatant was increased by 70 pg/mL (P < 0.05). Similar changes occurred in the PA group. After exposure to 4-HIL (5, 10, or 20 micromol/L) for 24 h, the glucose transportation was increased by 35%, 50%, and 60%, respectively. PCR results showed that along with increasing 4-HIL concentrations, the mRNA expression of cellular TNF-alpha showed a decreasing trend, showing statistical difference when compared with the model group and the PA group (P < 0.05). Compared with the model group, the TNF-alpha level in the supernatant was respectively reduced by 10 pg/mL, 18 pg/mL, and 39 pg/mL after intervention (P < 0.05).</p><p><b>CONCLUSION</b>4-HIL could remarkably improve high glucose-induced IR in 3T3-L1 adipocytes. Meanwhile, 4-HIL could inhibit the secretion of TNF-alpha.</p>

Overexpression of Corynebacterium glutamicum NAD kinase improves L-isoleucine biosynthesis / 生物工程学报

NAD kinase catalyzes the phosphorylation of coenzyme I [NAD(H)] to form coenzyme II [NADP(H)], and NADPH is an important cofactor in L-isoleucine biosynthesis. In order to improve NADPH supply, ppnK, the gene encoding NAD kinase in Corynebacterium glutamicum was cloned and separately expressed in an L-isoleucine synthetic strain, Brevibacterium lactofermentum JHI3-156, by an inducible expression vector pDXW-8 and a constitutive expression vector pDXW-9. Compared with the control strain JHI3-156/pDXW-8, NAD kinase activity of the inducible ppnK-expressing strain JHI3-156/pDXW-8-ppnK was increased by 83.5%. NADP(H)/NAD(H) ratio was also increased by 63.8%. L-isoleucine biosynthesis was improved by 82.9%. Compared with the control strain JHI3-156/pDXW-9, NAD kinase activity of the constitutive ppnK-expressing strain JHI3-156/pDXW-9-ppnK was increased by 220%. NADP(H)/ NAD(H) ratio and NADPH concentration were increased by 134% and 21.7%, respectively. L-isoleucine biosynthesis was increased by 41.7%. These results demonstrate that NAD kinase can improve the coenzyme II supply and L-isoleucine biosynthesis, which would also be useful for biosynthesis of other amino acids.

Polymorphism of M341I in the Type A Human Natriuretic Peptide Receptor Gene in Essential Hypertension in Korea / 이화의대지

OBJECTIVES: There are 3 subtypes of natriuretic peptide (NP) receptors: type A natriuretic peptide receptor (NPRA), NPRB, and NPRC. The NPRA gene polymorphism, consisting of substition of methionine (ATG) to isoleucine (ATC) at nucleotide 1023 (M341I) of exon 3 was revealed to be associated with increased risk for essential hypertension (EH) in Japanese people. The purpose of this study is to investigate association between EH and the M341I polymorphism in the NPRA gene in Korea. METHODS: Eighty patients in whom type B natriuretic peptide (BNP) was measured were enrolled in this study. 66 patients had EH and 14 patients did not. The polymorphism of M341I was evaluated by multiplex genotyping polymerase chain reaction and by sequencing analysis. RESULTS: The overall distribution of alleles was not significantly different between the control and EH groups. However, the C/C homozygous genotype was found only in the EH group. In the EH group, patient carrying the C/C homozygous genotype had the trend of having higher systolic and diastolic BP levels regardless of the previous treatment, even though other laboratory markers including BNP levels had no significant differences according to the genotypes. CONCLUSION: This would be meaningful for the first identification of the M341I polymorphism in the NPRA gene and for the first suggestion of association of the EH with it in Korea.

Crystal structure of the ubiquitin-like domain of human TBK1

TANK-binding kinase 1 (TBK1) is an important enzyme in the regulation of cellular antiviral effects. TBK1 regulates the activity of the interferon regulatory factors IRF3 and IRF7, thereby playing a key role in type I interferon (IFN) signaling pathways. The structure of TBK1 consists of an N-terminal kinase domain, a middle ubiquitin-like domain (ULD), and a C-terminal elongated helical domain. It has been reported that the ULD of TBK1 regulates kinase activity, playing an important role in signaling and mediating interactions with other molecules in the IFN pathway. In this study, we present the crystal structure of the ULD of human TBK1 and identify several conserved residues by multiple sequence alignment. We found that a hydrophobic patch in TBK1, containing residues Leu316, Ile353, and Val382, corresponding to the "Ile44 hydrophobic patch" observed in ubiquitin, was conserved in TBK1, IκB kinase epsilon (IKKɛ/IKKi), IκB kinase alpha (IKKα), and IκB kinase beta (IKKβ). In comparison with the structure of the IKKβ ULD domain of Xenopus laevis, we speculate that the Ile44 hydrophobic patch of TBK1 is present in an intramolecular binding surface between ULD and the C-terminal elongated helices. The varying surface charge distributions in the ULD domains of IKK and IKK-related kinases may be relevant to their specificity for specific partners.

Effect of Oral Supplementation with Branched-chain Amino Acid (BCAA) during Radiotherapy in Patients with Hepatocellular Carcinoma: A Double-Blind Randomized Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The present study evaluated whether oral supplementation with a branched-chain amino acid (BCAA) improves the biochemical and amino acid profiles of liver tumor patients undergoing radiotherapy. MATERIALS AND METHODS: Patients were randomly assigned to one of 2 groups: a group given oral supplementation with BCAA granules (LIVACT granules; Samil Pharm Co., Korea, each granule containing L-isoleucine 952 mg, L-leucine 1,904 mg, and L-valine 1,144 mg) during radiotherapy, or a placebo group. Physical and biochemical examinations and measurements, including subjective symptoms, Child-Pugh class, body mass index, plasma albumin concentration, and plasma amino acid profiles were monitored. RESULTS: Fifty were enrolled between November 2005 and November 2006. We also analyzed data from 37 hepatocellular carcinoma (HCC) patients in order to evaluate a more homogenous group. The two groups of patients were comparable in terms of age, gender, Child-Pugh score, and underlying hepatitis virus type. Serum albumin, total protein, liver enzymes, and cholesterol showed a tendency to increase in the BCAA group. In this group, the percentage of cases that reverted to normal serum albumin levels between 3 and 10 weeks after administration of BCAA was significantly higher (41.18%) than in the placebo group (p=0.043). CONCLUSION: Oral supplementation with a BCAA preparation seems to help HCC patients undergoing radiotherapy by increasing the BCAA concentration.

Effect of Oral Supplementation with Branched-chain Amino Acid (BCAA) during Radiotherapy in Patients with Hepatocellular Carcinoma: A Double-Blind Randomized Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The present study evaluated whether oral supplementation with a branched-chain amino acid (BCAA) improves the biochemical and amino acid profiles of liver tumor patients undergoing radiotherapy. MATERIALS AND METHODS: Patients were randomly assigned to one of 2 groups: a group given oral supplementation with BCAA granules (LIVACT granules; Samil Pharm Co., Korea, each granule containing L-isoleucine 952 mg, L-leucine 1,904 mg, and L-valine 1,144 mg) during radiotherapy, or a placebo group. Physical and biochemical examinations and measurements, including subjective symptoms, Child-Pugh class, body mass index, plasma albumin concentration, and plasma amino acid profiles were monitored. RESULTS: Fifty were enrolled between November 2005 and November 2006. We also analyzed data from 37 hepatocellular carcinoma (HCC) patients in order to evaluate a more homogenous group. The two groups of patients were comparable in terms of age, gender, Child-Pugh score, and underlying hepatitis virus type. Serum albumin, total protein, liver enzymes, and cholesterol showed a tendency to increase in the BCAA group. In this group, the percentage of cases that reverted to normal serum albumin levels between 3 and 10 weeks after administration of BCAA was significantly higher (41.18%) than in the placebo group (p=0.043). CONCLUSION: Oral supplementation with a BCAA preparation seems to help HCC patients undergoing radiotherapy by increasing the BCAA concentration.

Identification and quantitative determination of 4-hydroxyisoleucine in trigonella foenum-graecum L. from Iran

The seeds of Trigonella foenum-graecum L. [Fenugreek] are used in Iranian traditional medicine as blood sugar lowering agent. According to last investigations, hypoglycemic property is related to the amino acids of the plant specially 4-hydroxyisoleucine. In this research, identification and quantitative determination of 4-hydroxyisoleucine in Fenugreek seeds from Shahreza, central part of Iran, has been investigated. Methods: Powdered seeds of the plant were defatted and then extracted by using ethanol 50%. In order to isolation of 4-hydroxyisoleucine from the extract, ion exchange chromatography was used and the amino acid content was determined by using high performance liquid chromatography technique. The results showed the presence of 0.4% 4-hydroxyisoleucine in the plant seeds which is considerable amount of the amino acid in the plant collected from Iran. According to the effects of 4-hydroxyisoleucine such as antihyperglycemic and antihyperlipidemic properties, fenugreek can be considered as an herbal remedy in supportive treatment of these disorders

[Quantification of 4-hydroxyisoleucine content in treated and untreated fenugreek seeds using GC]

There are some reports regarding antidiabetic activity of 4-hydroxyisoleucine, however its hypoglycemic property must be proven completely before presentating as a new drug. Preparing suitable amount of 4-hydroxyisoleucine is nesessary for supporting in vivo studies. The purpose of the present study was isolation of 4-hydroxyisoleucine, comparing 4-hydroxyisoleucine content in two samples of Fenugreek seeds collected form two distinct parts of Iran and finally the effect of germination and supplementation by isoleucine on 4-hydroxyisoleucine yield. 4-hydroxyisoleucine was successfully isolated passing seed amino acid extract through ion exchange and silica gel column and then crystallization on methanol. 4- hydroxyisoleucine content in various samples was assessed by GC. Seeds from Northwestern of Iran had higher amount of 4-hydroxyisoleucine in comparison to Central Iran. Furthermore yields from germinating seeds were 2 folds higher than the dried seeds, and could be increased 6 fold by inclusion of isoleucine supplement in the irrigation water. simple procedures can be engaged for improving yields of 4-hydroxyisoleucine isolated from fenugreek up to six-fold by using of germinated seeds supplemented with isoleucine

Anti-hyperglycaemic activity of IND 01 and its interaction with glyburide and pioglitazone in alloxan induced diabetic mice

The antihyperglycaemic activity of IND 01 and its interaction with glyburide and pioglitazone on serum glucose, body weight and oral glucose tolerance test [OGTT] was determined in alloxan-induced diabetic mice. IND 01 [100 mg/kg], glyburide [10 mg/kg], pioglitzone [10 mg/kg] and their concomitant administration were administered orally in alloxan [80 mg/kg, i.v.] induced diabetic mice. The study design consisted of estimation of serum glucose after acute, subacute and glucose load administration. Administration of IND 01 [100 mg/kg] alone significantly [p<0.001] reduced serum glucose level at 6 h after administration. The antihyperglycaemic effect of glyburide and their concomitant administration of IND 01 with glyburide were similar, that is, onset was 2 h; peak effect was 6 h but the effect waned at 24 h. The onset of concomitant administration of IND 01 with pioglitazone was 4 h; peak effect was at 6 h but the effect waned at 24 h. In the subacute study, reduction in serum glucose was observed on 28[th] day after withdrawal for 7 days. The effects of concomitant administration were more pronounced than single drug treatment. In mice treated with either IND 01 [100 mg/kg], glyburide, pioglitazone alone or their combination, the body weight was not reduced in contrast to that in the control group. In the oral glucose tolerance test [OGTT], increased glucose utilization was observed in animals after concomitant administration of IND 01 [100 mg/kg] and glyburide [10 mg/kg] as well as IND 01 [100 mg/kg] and pioglitazone [10 mg/kg]. The concomitant administration of IND 01 with glyburide as well as pioglitzone produced synergistic antihyperglycaemic effect than either drug alone

[aroA gene sequencing of native Escherichia coli O78: k80 isolated from avian colibacillosis in Iran]

aroA is an important gene which produces aromatic amino acids essential for bacterial life. The sequence of this gene was shown to be different in bacteria, so the purpose of this study was sequencing of aroA gene in native E. coli O78: K80 to determine the putative differences between this strain and other E. coli O78: K80 strains in gene bank. PCR was used to amplify aroA gene in native E. coli O78: K80. The amplified 1.206 kb PCR product was extracted from Agaros gel, ligated in pTZ57R and sequenced. Blast analysis showed that aroA sequence in native E. coli is different from already submitted homolog gene in Gene Bank. Two amino acids were shown to be different from those already found in other E. coli strains. This different amino acids are Isoleucine and Glycine instead of 39[th] Threonine and 240[th] Glutamic acid, respectively. Knowing these differences are important for doing molecular techniques, designing primers and future studies

Enfermedad de orina a jarabe de arce / Maple syrup urine disease

Las enfermedades metabólicas pueden presentarse con síntomas, signos y laboratorios inespecíficos, que si no se consideran entre los diagnósticos diferenciales pueden retrasar el diagnóstico de estos pacientes, lo que lleva a un alto grado de secuelas neurológicas o muerte en etapas tempranas. La enfermedad de Orina a Jarabe de Arce es una enfermedad metabólica de baja incidencia caracterizada por la acumulación de niveles tóxicos de valina, isoleucina y principalmente leucina. Se presenta un paciente sin antecedentes que a los 11 días de vida comienza con mala actitud alimentaria, letargia y fontanela tensa. Descartadas las causas infectológicas se realizó un screening para enfermedades metabólicas. Se diagnosticó Leucinosis (Enfermedad de orina con olor a Jarabe de Arce) y se inició el tratamiento con restricción de leucina, valina e isoleucina en la dieta. A los pocos días del tratamiento el paciente mostró evidencias de mejoría clínica y en los parámetros de laboratorio.

Clinical signs, symptoms and lab tests of neonatal metabolic diseases may be unspecific and a high grade of suspicion is necessary to include them among the differential diagnosis avoiding a significant delay in recognizing this condition and consequent risk of neurologic handicap or early dead. Maple syrup urine disease is a congenital metabolic disorder with a low rate of prevalence and characterized by a toxic accumulation of the amino acids valine, isoleucine and mainly leucine. In this report we describe the history of a patient apparently healthy that on the 11th day after birth initiates symptoms like poor feeding, lethargy and tense fontanel. Excluded sepsis a work up for metabolic disease was performed, being diagnosed a leucinosis (Maple syrup urine disease). A dietary treatment with leucine, valine and isoleucine restriction was immediately initiated and a few days after the patient showed significant clinical and lab improvement. A short description and discussion of this disease is presented.

Thermodynamic study on the complexation of dioxovanadium [V] with isoleucine and glycylglycine

The dissociation constants of isoleucine and glycylglycine have been determined using a suitable computer program which employs a nonlinear least squares method. The complexation of dioxovanadium [V] with glycylglycine was studied by a combination of potentiometric and spectrophotometric techniques in aqueous solution at 25 °C and 0.1 mol dm[-3] ionic strength [NaClO[-4] in the pH range 2.00 to 2.60. All of the calculations have been done by the computer program Excel 2000. Dioxovanadium [V] complexes formed by isoleucine in the pH range 1.30 to 2.5 and between t = 25 °C and 35 °C with one mole dm[-3] sodium perchlorate as the ionic medium, have been investigated potentiometrically and spectrophotometrically. Ultimately, the thermodynamic functions [delta G°, delta H°, delta S°] have been calculated for the complexation of dioxovanadium [V] with isoleucine