Efficacy of nutrition therapy with food rich in methionine for treating nonalcoholic fatty liver / Eficacia de la terapia nutricional con alimentos ricos en metionina para el tratamiento de hígado graso no alcohólico

Non-Alcoholic Fatty Liver disease (NAFLD) can lead to Non Alcoholic steatohepatitis (NASH), cirrhosis, and liver cancer. The treatment for NAFLD involves modification of caloric intake and physical activity. NAFLD has a pro-oxidant nature; therefore, it is logical to suppose that the antioxidant methionine can be used as a treatment for this disease. Aim. This study aimed to evaluate the effect of high-methionine dietary therapy on patients with NAFLD. Materials and methods. A randomized clinical study was conducted over three months. In this study, 121 NAFLD patients participated, and the age of the participants was ≥ 20 years (experimental group included 56 and control group 65), all of whom were randomized and matched by sex, recluted from the ISSSTE hospital in Xalapa, Mexico. The patients were instructed to consume food to cover the recommended methionine daily doses, and the daily amount consumed was calculated. Methionine effect was measured as NAFLD regression and quality of life improvement. Results. Nutritional therapy induced NAFLD regression and diminished central fat accumulation, blood pressure, and the fatty liver index. Some parameters, such as liver enzymes, did not changed. The quality of life of patients improved after treatment. Conclusions. In this study, we show a hepatoprotective effect induced only in three months of chances in the diet, thus, a longer diet may generate more relevant benefits in the resistant parameters of our study(AU)

La enfermedad del hígado graso no alcohólico (NAFLD) puede conducir a la esteatohepatitis no alcohólica (NASH), la cirrosis y el cáncer de hígado. El tratamiento para NAFLD es la modificación de la ingesta calórica y la actividad física. Debido a que NAFLD tiene una naturaleza pro-oxidante; es lógico suponer que el antioxidante metionina puede utilizarse en el tratamiento de esta enfermedad. Objetivo. el presente trabajo evaluó el papel de la terapia nutricional con alimentos ricos en metioninaen pacientes con NAFLD. Materiales y Métodos. Se realizó un ensayo clínico aleatorizado durante tres meses. Participaron en el estudio 121 pacientes con NAFLD con edad ≥ 20 años (56 en el grupo experimental y 65 en el control), todos aleatorizados y pareados por sexo, reclutados de la Clínica Hospital ISSTE en la ciudad de Xalapa, México, en el año 2015. Se instruyó a los pacientes en consumir los alimentos hasta completar la dosis diaria recomendada de metioninay se calculó la cantidad diaria consumida. Su efecto se midió como la regresión de NAFLD y la mejora de la calidad de vida. Resultados. La terapia nutricional retrocedió NAFLD; disminuyó la acumulación de grasa central, la presión arterial y el índice de hígado graso. Algunos parámetros, como las enzimas de la función hepática, no se modificaron con el tratamiento. Otro parámetro fue la mejora de la calidad de vida de los pacientes tratados. Conclusiones. En este trabajo mostramos un impacto hepatoprotector producido con tan solo tres meses de cambios en la dieta, por lo que una dieta más prolongada podría generar beneficios aún más significativos en los parámetros resistentes en nuestro protocolo(AU)

Microbial production of S-adenosyl-l-methionine: a review / 生物工程学报

S-adenosyl-l-methionine (SAM) is ubiquitous in living organisms and plays important roles in transmethylation, transsulfuration and transamination in organisms. Due to its important physiological functions, production of SAM has attracted increasing attentions. Currently, researches on SAM production mainly focus on microbial fermentation, which is more cost-effective than that of the chemical synthesis and the enzyme catalysis, thus easier to achieve commercial production. With the rapid growth in SAM demand, interests in improving SAM production by developing SAM hyper-producing microorganisms aroused. The main strategies for improving SAM productivity of microorganisms include conventional breeding and metabolic engineering. This review summarizes the recent research progress in improving microbial SAM productivity to facilitate further improving SAM productivity. The bottlenecks in SAM biosynthesis and the solutions were also addressed.

Comparative analysis of various sources of selenium on the growth performance and antioxidant status in broilers under heat stress / Análise comparativa de várias fontes de selênio no desempenho de crescimento e status antioxidante em frangos de corte sob estresse térmico

The supplementation of Selenium-enriched probiotics is effective in reducing oxidative stress and maintaining meat quality stability in broiler chicken especially under heat stress. An experimental study was conducted to perform Comparative analysis of Selenium yeast with inorganic Se in broilers under heat stress. A total of 120 broilers chicks of one day were assigned to 4 groups each consisting 30 chicks fed on same basal diet but different selenium sources. The basal diet of group D1 was not supplemented with Se source (Negative control), group D2 basal diet was supplemented with inorganic selenium (Sodium selenite 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase), group D3 basal diet was supplemented with commercially available organic selenium (Seleno-methionine 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase) and group D4 basal diet was supplemented with self-developed organic selenium (Se-enriched yeast 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase). The performance parameters i.e. feed intake (FI), live body weight (BW) and FCR were not significantly (p>0.05) effected by selenium supplementation in the starter phase but were significantly (p<0.05) effected in the finisher phase. Selenium supplementation significantly (p<0.05) effected serum Se level in different supplemented groups. Higher serum Se value (58.20±0.06) was recorded in D4 group. Similarly significantly lower selenium value was recorded for D4 and higher was recorded for D1 (11.36±0.08). However lower serum Paraoxonase (PON) value was recorded for D4 (13.24±0.01) and higher for D1 (13.33±0.03). Comparatively self-developed Se enriched yeast increased the Se accumulation and improved antioxidant system. Glutathione peroxidase (GPx) was found higher in D4 (12.333±0.03) followed by D3, D2 and D1 respectively. Whereas superoxide dismutase (SOD) was significantly lower (p<0.05) in D4 (0.1437±0.003) followed by D3 (0.1457±0.002). Selenium supplementation increased the [...].

A suplementação de probióticos enriquecidos com selênio é eficaz na redução do estresse oxidativo e na manutenção da estabilidade da qualidade da carne em frangos de corte, especialmente sob estresse por calor. Um estudo experimental foi conduzido para realizar uma análise comparativa da levedura selênio com o Se inorgânico em frangos de corte sob estresse térmico. Um total de 120 pintos de um dia foi dividido em 4 grupos, cada um consistindo de 30 pintos alimentados com a mesma dieta basal, mas com diferentes fontes de selênio. A dieta basal do grupo D1 não foi suplementada com fonte de Se (controle negativo), a dieta basal do grupo D2 foi suplementada com selênio inorgânico (selenito de sódio 0,22 mg / kg fase inicial e 0,15 mg / kg fase finalizadora), a dieta basal do grupo D3 foi suplementada com selênio orgânico disponível comercialmente (fase inicial de seleno-metionina 0,22 mg / kg e fase finalizadora de 0,15 mg / kg) e a dieta basal do grupo D4 foi suplementada com selênio orgânico autodesenvolvido (fermento enriquecido com Se 0,22 mg / kg fase inicial e 0,15 mg / kg fase finalizadora). Os parâmetros de desempenho, ou seja, consumo de ração (FI), peso corporal vivo (PC) e FCR não foram significativamente (p > 0,05) afetados pela suplementação de selênio na fase inicial, mas foram significativamente (p < 0,05) afetados na fase final. A suplementação de selênio afetou significativamente (p < 0,05) o nível de Se sérico em diferentes grupos suplementados. O maior valor de Se sérico (58,20 ± 0,06) foi registrado no grupo D4. Da mesma forma, valor de selênio significativamente menor foi registrado para D4 e maior foi registrado para D1 (11,36 ± 0,08). No entanto, um valor mais baixo de Paraoxonase (PON) sérica foi registrado para D4 (13,24 ± 0,01) e mais alto para D1 (13,33 ± 0,03). A levedura enriquecida com Se comparativamente autodesenvolvida aumentou o acúmulo de Se e melhorou o sistema [...].

Comparative analysis of various sources of selenium on the growth performance and antioxidant status in broilers under heat stress

Abstract The supplementation of Selenium-enriched probiotics is effective in reducing oxidative stress and maintaining meat quality stability in broiler chicken especially under heat stress. An experimental study was conducted to perform Comparative analysis of Selenium yeast with inorganic Se in broilers under heat stress. A total of 120 broilers chicks of one day were assigned to 4 groups each consisting 30 chicks fed on same basal diet but different selenium sources. The basal diet of group D1 was not supplemented with Se source (Negative control), group D2 basal diet was supplemented with inorganic selenium (Sodium selenite 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase), group D3 basal diet was supplemented with commercially available organic selenium (Seleno-methionine 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase) and group D4 basal diet was supplemented with self-developed organic selenium (Se-enriched yeast 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase). The performance parameters i.e. feed intake (FI), live body weight (BW) and FCR were not significantly (p>0.05) effected by selenium supplementation in the starter phase but were significantly (p 0.05) effected in the finisher phase. Selenium supplementation significantly (p 0.05) effected serum Se level in different supplemented groups. Higher serum Se value (58.20±0.06) was recorded in D4 group. Similarly significantly lower selenium value was recorded for D4 and higher was recorded for D1 (11.36±0.08). However lower serum Paraoxonase (PON) value was recorded for D4 (13.24±0.01) and higher for D1 (13.33±0.03). Comparatively self-developed Se enriched yeast increased the Se accumulation and improved antioxidant system. Glutathione peroxidase (GPx) was found higher in D4 (12.333±0.03) followed by D3, D2 and D1 respectively. Whereas superoxide dismutase (SOD) was significantly lower (p 0.05) in D4 (0.1437±0.003) followed by D3 (0.1457±0.002). Selenium supplementation increased the birds survival rate. Birds fed on Se enriched yeast showed higher Se deposition and better antioxidant capacity as compared to other sources of selenium. Se-enriched yeast displayed an improved result on Se deposition in tissues, and oxidative capacity, meat tenderness and immune response level as compared to other sources of selenium.

Resumo A suplementação de probióticos enriquecidos com selênio é eficaz na redução do estresse oxidativo e na manutenção da estabilidade da qualidade da carne em frangos de corte, especialmente sob estresse por calor. Um estudo experimental foi conduzido para realizar uma análise comparativa da levedura selênio com o Se inorgânico em frangos de corte sob estresse térmico. Um total de 120 pintos de um dia foi dividido em 4 grupos, cada um consistindo de 30 pintos alimentados com a mesma dieta basal, mas com diferentes fontes de selênio. A dieta basal do grupo D1 não foi suplementada com fonte de Se (controle negativo), a dieta basal do grupo D2 foi suplementada com selênio inorgânico (selenito de sódio 0,22 mg / kg fase inicial e 0,15 mg / kg fase finalizadora), a dieta basal do grupo D3 foi suplementada com selênio orgânico disponível comercialmente (fase inicial de seleno-metionina 0,22 mg / kg e fase finalizadora de 0,15 mg / kg) e a dieta basal do grupo D4 foi suplementada com selênio orgânico autodesenvolvido (fermento enriquecido com Se 0,22 mg / kg fase inicial e 0,15 mg / kg fase finalizadora). Os parâmetros de desempenho, ou seja, consumo de ração (FI), peso corporal vivo (PC) e FCR não foram significativamente (p > 0,05) afetados pela suplementação de selênio na fase inicial, mas foram significativamente (p 0,05) afetados na fase final. A suplementação de selênio afetou significativamente (p 0,05) o nível de Se sérico em diferentes grupos suplementados. O maior valor de Se sérico (58,20 ± 0,06) foi registrado no grupo D4. Da mesma forma, valor de selênio significativamente menor foi registrado para D4 e maior foi registrado para D1 (11,36 ± 0,08). No entanto, um valor mais baixo de Paraoxonase (PON) sérica foi registrado para D4 (13,24 ± 0,01) e mais alto para D1 (13,33 ± 0,03). A levedura enriquecida com Se comparativamente autodesenvolvida aumentou o acúmulo de Se e melhorou o sistema antioxidante. A glutationa peroxidase (GPx) foi encontrada maior em D4 (12,333 ± 0,03) seguido por D3, D2 e D1 respectivamente. Enquanto a superóxido dismutase (SOD) foi significativamente menor (p 0,05) em D4 (0,1437 ± 0,003) seguido por D3 (0,1457 ± 0,002). A suplementação de selênio aumentou a taxa de sobrevivência da ave. Aves alimentadas com levedura enriquecida com Se apresentaram maior deposição de Se e melhor capacidade antioxidante em comparação com outras fontes de selênio. A levedura enriquecida com Se apresentou um resultado melhorado na deposição de Se nos tecidos, capacidade oxidativa, maciez da carne e nível de resposta imune em comparação com outras fontes de selênio.

Comparative analysis of various sources of selenium on the growth performance and antioxidant status in broilers under heat stress / Análise comparativa de várias fontes de selênio no desempenho de crescimento e status antioxidante em frangos de corte sob estresse térmico

Abstract The supplementation of Selenium-enriched probiotics is effective in reducing oxidative stress and maintaining meat quality stability in broiler chicken especially under heat stress. An experimental study was conducted to perform Comparative analysis of Selenium yeast with inorganic Se in broilers under heat stress. A total of 120 broilers chicks of one day were assigned to 4 groups each consisting 30 chicks fed on same basal diet but different selenium sources. The basal diet of group D1 was not supplemented with Se source (Negative control), group D2 basal diet was supplemented with inorganic selenium (Sodium selenite 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase), group D3 basal diet was supplemented with commercially available organic selenium (Seleno-methionine 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase) and group D4 basal diet was supplemented with self-developed organic selenium (Se-enriched yeast 0.22mg/Kg starter phase and 0.15mg/Kg finisher phase). The performance parameters i.e. feed intake (FI), live body weight (BW) and FCR were not significantly (p>0.05) effected by selenium supplementation in the starter phase but were significantly (p<0.05) effected in the finisher phase. Selenium supplementation significantly (p<0.05) effected serum Se level in different supplemented groups. Higher serum Se value (58.20±0.06) was recorded in D4 group. Similarly significantly lower selenium value was recorded for D4 and higher was recorded for D1 (11.36±0.08). However lower serum Paraoxonase (PON) value was recorded for D4 (13.24±0.01) and higher for D1 (13.33±0.03). Comparatively self-developed Se enriched yeast increased the Se accumulation and improved antioxidant system. Glutathione peroxidase (GPx) was found higher in D4 (12.333±0.03) followed by D3, D2 and D1 respectively. Whereas superoxide dismutase (SOD) was significantly lower (p<0.05) in D4 (0.1437±0.003) followed by D3 (0.1457±0.002). Selenium supplementation increased the bird's survival rate. Birds fed on Se enriched yeast showed higher Se deposition and better antioxidant capacity as compared to other sources of selenium. Se-enriched yeast displayed an improved result on Se deposition in tissues, and oxidative capacity, meat tenderness and immune response level as compared to other sources of selenium.

Resumo A suplementação de probióticos enriquecidos com selênio é eficaz na redução do estresse oxidativo e na manutenção da estabilidade da qualidade da carne em frangos de corte, especialmente sob estresse por calor. Um estudo experimental foi conduzido para realizar uma análise comparativa da levedura selênio com o Se inorgânico em frangos de corte sob estresse térmico. Um total de 120 pintos de um dia foi dividido em 4 grupos, cada um consistindo de 30 pintos alimentados com a mesma dieta basal, mas com diferentes fontes de selênio. A dieta basal do grupo D1 não foi suplementada com fonte de Se (controle negativo), a dieta basal do grupo D2 foi suplementada com selênio inorgânico (selenito de sódio 0,22 mg / kg fase inicial e 0,15 mg / kg fase finalizadora), a dieta basal do grupo D3 foi suplementada com selênio orgânico disponível comercialmente (fase inicial de seleno-metionina 0,22 mg / kg e fase finalizadora de 0,15 mg / kg) e a dieta basal do grupo D4 foi suplementada com selênio orgânico autodesenvolvido (fermento enriquecido com Se 0,22 mg / kg fase inicial e 0,15 mg / kg fase finalizadora). Os parâmetros de desempenho, ou seja, consumo de ração (FI), peso corporal vivo (PC) e FCR não foram significativamente (p > 0,05) afetados pela suplementação de selênio na fase inicial, mas foram significativamente (p < 0,05) afetados na fase final. A suplementação de selênio afetou significativamente (p < 0,05) o nível de Se sérico em diferentes grupos suplementados. O maior valor de Se sérico (58,20 ± 0,06) foi registrado no grupo D4. Da mesma forma, valor de selênio significativamente menor foi registrado para D4 e maior foi registrado para D1 (11,36 ± 0,08). No entanto, um valor mais baixo de Paraoxonase (PON) sérica foi registrado para D4 (13,24 ± 0,01) e mais alto para D1 (13,33 ± 0,03). A levedura enriquecida com Se comparativamente autodesenvolvida aumentou o acúmulo de Se e melhorou o sistema antioxidante. A glutationa peroxidase (GPx) foi encontrada maior em D4 (12,333 ± 0,03) seguido por D3, D2 e ​​D1 respectivamente. Enquanto a superóxido dismutase (SOD) foi significativamente menor (p < 0,05) em D4 (0,1437 ± 0,003) seguido por D3 (0,1457 ± 0,002). A suplementação de selênio aumentou a taxa de sobrevivência da ave. Aves alimentadas com levedura enriquecida com Se apresentaram maior deposição de Se e melhor capacidade antioxidante em comparação com outras fontes de selênio. A levedura enriquecida com Se apresentou um resultado melhorado na deposição de Se nos tecidos, capacidade oxidativa, maciez da carne e nível de resposta imune em comparação com outras fontes de selênio.

The Effect of Dietary Supplement on Glutathione Level in Rats

Background: Oxidative stress occurs due to decreased glutathione inside the body. Some supplements may promote and stimulate glutathione production in the liver. This article aims to investigate the impact of different supplements on enhancing glutathione synthesis in rats’ livers. For this purpose 42 rats (male albino) were separated into 7 groups, each including 6 animals with average weights ranging between 150 and 160 g. Group 1 (control) and different groups consumed a basal diet for 8 weeks, whereas group 2 received 500 mg/kg bw of L-cysteine daily. Group 3 received 250 mg/kg bw of methionine, while group 4 got 250 mg/kg of L cysteine plus 125 mg/kg of methionine daily. Spirulina (20 mg/kg bw), turmeric (500 mg/kg bw), and dried garlic (500 mg/kg bw), respectively, were given to groups 5, 6, and 7. Results: Utilizing the various dietary supplements decreased levels of liver function enzymes, bilirubin, urea, creatinine, and malondialdehyde while enhancing levels antioxidant enzymes of liver, and increased glutathione of kidneys and liver. However, cysteine alone at 500 mg/kg bw decreased glutathione formation in the liver and kidneys. Compared to the amino acid supplements (group 2, 3, 4) used, spirulina, turmeric, and dried garlic had a significant impact on reducing liver function enzymes, bilirubin, uric acid, creatinine, urea, and malondialdehyde and increasing antioxidant enzymes, and glutathione while turmeric supplement showed the best influence. Using dietary supplements did not result in any pathological modifications in the liver tissues, but there were some unsatisfactory minor alterations. However, group 2 showed considerable pathological developments in the liver tissues. Conclusion: According to the findings, using the suggested dietary supplement except for cysteine alone can promote and encourage glutathione synthesis in different organs, especially the liver, hence alleviating the effects of oxidative stress associated with several illnesses.

D-methionine eradicates Porphyromonas gingivalis biofilms by suppressing the cyclic di-GMP level / 口腔疾病防治

Objective@#To investigate the mechanisms by which D-methionine (D-Met) eradicates Porphyromonas gingivalis biofilms by suppressing cyclic dimeric GMP (c-di-GMP) levels.@*Methods @#Cell viability, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured to determine the effective concentrations of D-Met, which were subsequently used in the following experiments. During the P. gingivalis biofilm formation inhibition experiment and the mature biofilm disassembly experiment, biofilm biomass, exopolysaccharide (EPS), biofilm morphology, integrity of the cell membrane, and the level of c-di-GMP were determined. @*Results @# D-Met < 40 mmol/L was biocompatible. During the biofilm formation inhibition and mature biofilm disassembly experiments, D-Met ≥ 20 mmol/L decreased the biofilm biomass and the production of EPS. SEM analysis showed that the extracellular matrix and bacterial density were drastically reduced by D-Met ≥ 20 mmol/L. TEM detection showed that 35 mmol/L D-Met ruptured the cell membrane during biofilm formation and increased the permeability of the cell membrane in the disassembly phase of mature biofilms. C-di-GMP levels decreased with increasing concentrations of D-Met in a concentration-dependent manner.@* Conclusion @# D-Met ≥ 20 mmol/L could eradicate P. gingivalis biofilms by suppressing c-di-GMP levels.

Involvement of Opioid Peptides in the Analgesic Effect of Spinal Cord Stimulation in a Rat Model of Neuropathic Pain / 神经科学通报·英文版

Spinal cord stimulation (SCS)-induced analgesia was characterized, and its underlying mechanisms were examined in a spared nerve injury model of neuropathic pain in rats. The analgesic effect of SCS with moderate mechanical hypersensitivity was increased with increasing stimulation intensity between the 20% and 80% motor thresholds. Various frequencies (2, 15, 50, 100, 10000 Hz, and 2/100 Hz dense-dispersed) of SCS were similarly effective. SCS-induced analgesia was maintained without tolerance within 24 h of continuous stimulation. SCS at 2 Hz significantly increased methionine enkephalin content in the cerebrospinal fluid. The analgesic effect of 2 Hz was abolished by μ or κ opioid receptor antagonist. The effect of 100 Hz was prevented by a κ antagonist, and that of 10 kHz was blocked by any of the μ, δ, or κ receptor antagonists, suggesting that the analgesic effect of SCS at different frequencies is mediated by different endorphins and opioid receptors.

Identification and functional analysis of combined oxidative phosphorylation deficiency 28 gene mutation / 中华内科杂志

Objective:To report a case of combined oxidative phosphorylation deficiency 28 (COXPD28) in China, identified the pathogenic mutation and explored the pathogenic mechanism preliminarily.Methods:The clinical characteristics of a patient with COXPD28 were retrospectively analyzed and the pathogenic mutations were identified by mitochondrial gene sequencing and whole exome sequencing. The wild-type and mutant plasmids of pathogenic genes were constructed, and effect of mutation on protein expression by quantitative real-time PCR (qPCR) and Western blot were evaluated. Statistical methods mainly used one-way ANOVA and LSD test.Results:A 21 year old female patient presented with lactic acid poisoning due to repeated chest distress and wheezing since childhood. The sequencing of the whole exon group gene found that solute carrier family 25 member 26 (SLC25A26) gene had a compound heterozygous mutation (c.34G>C, p.A12P; c.197C>A, p.A66E), which was the first report in China. In vitro function test showed that the expression levels of SLC25A26 mRNA and S-adenosylmethionine carrier (SAMC) protein in cells transfected with SLC25A26 mutant plasmid were significantly lower than those transfected with wild type plasmid. The p.A66E mutant plasmid reduced the expression level of SLC25A26 mRNA and SAMC protein to 6% and 26% of wild type plasmids respectively (both P<0.001), while p.A12P mutant plasmid decreased to 62% and 82% of wild type plasmids respectively ( P<0.001, P=0.044). When the double mutant (p.A66E+p.A12P) plasmids were co-transfected, the expression levels of SLC25A26 mRNA and SAMC protein decreased to 47% and 57% of the wild type plasmids, respectively ( P<0.001, P=0.001). Conclusion:The pathogenic mutation gene of this patient with COXPD28 is SLC25A26 gene mutation (p.A66E, p.A12P), which causes the decrease of SLC25A26 expression level, mitochondrial oxidative phosphorylation dysfunction, and induces COXPD28.

Assessment of MTR Rs1805087 SNP as Possible Modifier ofSickle Cell Disease Severity in a Nigerian Population

BACKGROUND: Sickle cell disease is the commonest geneticdisorder in Nigeria, affecting 2­3% of an estimated population of 160million people. The role of genetic mutations in folate cycle genes,and the variable phenotypic expressions constituting disease severity,needs to be critically examined.OBJECTIVE: This study was carried out to establish the pattern ofmethionine synthase gene mutations (rs1805087 SNP), and its possibleassociation with disease severity in adults with sickle cell anaemia inLagos, Nigeria.METHODOLOGY: This is a cross-sectional study of seventy (70)subjects with sickle cell disease (HbSS) matched for age and genderwith known apparently healthy haemoglobin genotype AA (HbAA)subjects, as cases and controls respectively. Structured questionnaireswere used to obtain demographic, clinical and other phenotypic dataneeded to compute disease severity. Pattern of MTR A2756G genemutation and homocysteine assay (Hcy) were assessed by PolymeraseCh ain Reaction and Enzyme- linked Immun osorbent Assayrespectively. Full blood count analysis of participants was done usingthe KX-21 Automated Analyzer (Sysmex Corporation, Japan).RESULTS: The mutant genotypes MTR 2756 AG/GG were recordedin 46.4% (n =55) of subjects with disease severity score >7. Elevatedplasma homocysteine (HHcy) was significantly associated withdisease severity among HbSS subjects (OR=17.2, CI: 3.490-86.079;p=0.0001). Conversely, no significant association was observed withthe mutant genotypes MTR 2756 AG/GG and disease severity(p>0.05).CONCLUSION: While HHcy is significantly associated withphenotypic expression of HbSS, the MTR 2756 SNPs did not appearto independently influence homocysteine level or disease severity inHbSS subjects

Mechanism of Tibetan medicine Ershiwuwei Songshi Pills in regulating intestinal flora and improving non-alcoholic steatohepatitis / 中国中药杂志

This study aimed to investigate the effect of Tibetan medicine Ershiwuwei Songshi Pills(ESP) on the intestinal flora of non-alcoholic steatohepatitis(NASH) mice. Forty-eight male C57 BL/6 mice were randomly divided into the control group, model(methionine-choline-deficient, MCD) group, high-(0.8 g·kg~(-1)), medium-(0.4 g·kg~(-1)), and low-dose(0.2 g·kg~(-1)) ESP groups, and pioglitazone(PGZ, 10 mg·kg~(-1)) group, with eight mice in each group. Mice in the control group were fed with normal diet, while those in the remaining five groups with MCD diet for five weeks for inducing NASH. During modeling, they were gavaged with the corresponding drugs. The changes in body mass, daily water intake, and daily food intake were recorded. At the end of the experiment, the liver tissues were collected and stained with hematoxylin-eosin(HE) for observing the pathological changes, followed by oil red O staining for observing fat accumulation in the liver. The levels of serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) and triglyceride(TG) in liver tissue were measured. The changes in intestinal flora of mice were determined using 16 S rRNA high-throughput sequencing technology. The results showed that compared with the model group, the high-, medium-and low-dose ESP groups and the PGZ group exhibited significantly lowered AST and ALT in serum and TG in liver tissues and alleviated hepatocellular steatosis and fat accumulation in the liver. As demonstrated by 16 S rRNA sequencing, the abundance index and diversity of intestinal flora decreased in the model group, while those increased in the ESP groups. Besides, the Firmicutes to Bacteroidetes ratio decreased at the phylum level. In the alteration of the composition of intestinal flora, ESP reduced the abundance of Erysipelotrichia and Faecalibaculum but increased the abundance of Desulfovibrionaceae, Rikenellaceae, Lachnospiraceae, and Ruminococcaceae. This study has revealed that ESP has a protective effect against NASH induced by MCD diet, which may be related to its regulation of the changes in intestinal flora, alteration of the composition of intestinal flora, and inhibition of the intestinal dysbiosis.

Oxidized related substance of methionine in compound amino acid injection (18AA-Ⅱ) / 中国药科大学学报

@#TA method for the content determination of methionine sulfoxide and methionine sulfone in compound amino acid injection (18AA-II) was established in order to investigate their level in 155 batches of this product, and to explore the reason for the generation of these two impurities.The determination was performed on an Agilent Poroshell 120 EC-C18 column with mobile phases of sodium acetate/tetrahydrofuran solution (A) and sodium acetate solution -acetonitrile-methanol (B, 200∶400∶400) (gradient elution) at the flow rate of 0.5 mL/min.The excitation wavelength and the emission wavelength of the fluorescence detector were 233 nm and 441 nm, respectively.The column temperature was 40 °C, and the injection volume was 8 μL.The contents of methionine sulfoxide and methionine sulfone from 155 batches of compound amino acid injection (18AA-II) was determined using this method, and the residual oxygen content was detected by headspace gas analyzer.The results showed that the linear range of methionine sulfoxide and methionine sulfone were 0.128 1-10.250 0 μg/mL (r = 0.999 9) and 0.261 0-10.440 0 μg/mL (r = 0.999 8), respectively.The limits of quantitation were 0.13 μg/mL and 0.26 μg/mL, respectively; the limits of detection were 0.04 μg/mL and 0.09 μg/mL, respectively.RSDs of precision, stability and repetitive test were all lower than 1.3%.The recoveries ranged 98.00%-100.79% (RSD = 1.15%, n = 9) and 98.19%-102.31% (RSD = 1.33%, n = 9).The content level of oxidized related substances from different manufacturers showed significant difference, showing relevance with the residual oxygen content to some extent, yet no significant correlation with the added amount of antioxygen (sodium pyrosulfite).The method is validated to be useful for the content control of methionine sulfoxide and methionine sulfone in compound amino acid injection (18AA-II).It is quite necessary to include the determination of oxidized related substance into the quality specification.Manufacturers should strengthen the control of remaining oxygen in their products.

Intervention Effect and Mechanism of Celastrol on NASH Induced by Methionine-choline Deficiency Diet in Mice / 中国药房

OBJECTIVE：To study the intervention eff ects and pot ential m echanism of celastrol on non-alcoholic steatohepatitis （NASH）induced by methionine-choline deficiency （MCD）diet. METHODS ：Male C 57BL/6J mice were randomly divided into normal control group ，model group ，celastrol low-dose and high-dose groups [ 0.5，1 mg/（kg·d）]，with 7 mice in each group. The normal control group was given a methionine-choline sufficient diet ，while the model group and administration groups were fed an MCD diet to induce NASH model. At the same time ，normal control group and model group were given polyoxyethylene castor oil intragastrically；administration groups were given relevant drugs intragastrically ；the volume of gavage was 0.1 mL/g，once a day ， for consecutive 4 weeks. The liver morphology was observed ，and the pathological changes of liver tissue were observed by HE staining and oil red O staining. The levels of serum liver enzymes （AST，ALT），and the levels of lipid indexes （TC，TG）in serum and liver tissue were detected by enzyme method. The protein expression of NF-κB p65，TNF-α and IL-6 in liver tissue were determined by Western blotting assay. RESULTS ：Compared with normal control group ，the volume of the liver was reduced and the color was yellow ，and the surface was rough in model group ；inflammatory cell infiltration ，fat vacuoles and lipid droplets aggregation were found in the liver tissue ；the serum levels of TC and TG were significantly decreased ，the levels of serum liver enzymes and protein expression of NF-κB p65，TNF-α and IL-6 in liver tissue were significantly increased （P＜0.01）. Compared with model group ，the liver surface of each administration group was ruddy and smooth without brown spots ，the inflammatory cells and fat vacuoles in liver tissue were reduced ，and the coverage area of lipid droplets was reduced ；the levels of serum TC and TG were significantly increased ，the levels of serum liver enzymes ，the levels of TG and protein expression of NF-κB，TNF-α and IL-6（except for celastrol low-dose group ）in liver tissue were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ： Celastrol can improve the liver injury of NASH model mice induced by MCD diet ，which is related to the reduction of TG accumulation in liver tissue and inhibition of the expression of inflammatory related factors.

Identification of geographical origins of Cordyceps based on data of amino acids with self-organizing map neural network / 中国中药杂志

In this study, data of amino acids of Cordyceps samples from Qinghai and Tibet was analyzed with self-organizing map neural network. A model of XY-Fused network was established with the content of 8 major amino acids and total amino acids for the identification of geographical origins of Cordyceps from Qinghai and Tibet. It had the prediction accuracy of 83.3% for the test set. In addition, data mining indicated that methionine was a special kind of amino acid in Cordyceps which could serve as a marker to identify its geographical origins. On this basis, the content ratio of methionine to total amino acids was proposed to be a quantifiable indicator to distinguish Cordyceps from Qinghai and Tibet.

Effectiveness and optimum level of protected methionine in fattening pig diets / Efectividad y nivel óptimo de metionina protegida en dietas para cerdos en engorde / Eficácia e nível ideal de metionina protegida em dietas para suínos de engorda

Abstract Background: Methionine (Met) requirements have not been clearly established for fattening pigs due to their metabolic interrelationships and its bioavailability for protein synthesis. Objective: To determine the optimum level of regular crystalline or protected Met in pig diets from nursery to finishing. Methods: A total of 48 crossbred pigs (11.74±1.72 kg of initial body weight) were used. The treatments consisted of adding four levels (0.00, 0.05, 0.10, 0.15% in addition to dietary content) and two types of Met (regular and protected) to pig diets. Results: Nursery, Finishing I, and II pigs fed protected Met increased daily feed intake (DFI; p≤0.10). Protected Met raised daily weight gain (DWG) in nursery pigs and increased backfat thickness (BT) in nursery and grower pigs (p≤0.10). In Finishing I pigs, protected Met increased DWG and improved carcass characteristics (p≤0.10). In nursery and grower pigs, an extra 0.15% Met decreased feed:gain ratio (FGR; p≤0.10). In grower and Finishing II pigs fed extra 0.05% Met improved DWG and extra 0.10% Met reduced plasma urea concentration (p≤0.10). Conclusions: Feeding protected Met in pig diets increases DWG, DFI and BT. Increasing 0.05-0.15% Met level improves FGR, DWG, potentially reducing nitrogen excretion to the environment.

Resumen Antecedentes: los requerimientos de metionina (Met) para cerdos en crecimiento no han sido claramente establecidos, lo que se debe a sus relaciones metabólicas y su biodisponibilidad para la síntesis proteica. Objetivo: determinar el nivel óptimo de Met regular o protegida en dietas para cerdos en crecimiento. Métodos: Se utilizaron un total de 48 cerdos híbridos (11,74±1,72 kg peso vivo inicial). Los tratamientos consistieron en niveles incrementales (0,00, 0,05, 0,10, 0,15% adicionales al contenido de la dieta) y dos tipos de Met (regular y protegida) en la dieta. Resultados: los cerdos en iniciación, Finalización I, y II, alimentados con Met protegida tuvieron un mayor consumo diario de alimento (DFI; p≤0,10). La Met protegida aumentó la ganancia diaria de peso (DWG) durante la etapa de iniciación, e incrementó el grosor de la grasa dorsal (BT) en iniciación y levante (p≤0,10). Durante Finalización I, la Met protegida aumentó la DWG y mejoró las características de la canal (p≤0,10). Durante iniciación y levante, 0,15% extra de Met disminuyó la conversión alimenticia (FGR; p≤0,10). Los cerdos en levante y Finalización II alimentados con 0,05% extra de Met mejoraron la DWG y con 0.10% extra de Met redujeron la concentración de urea en plasma (p≤0,10). Conclusiones: el uso de Met protegida incrementa DWG, DFI y BT. El aumento del nivel de Met de 0,05-0,15% mejora FGR y DWG, y podria disminuir la excreción de nitrógeno al ambiente.

Resumo Antecedentes: os requisitos de metionina (Met) para suínos de engorda não foram claramente estabelecidos devido às suas relações metabólicas e sua biodisponibilidade para a síntese de proteínas. Objetivo: determinar o nível ideal de Met regular ou protegida em dietas para suínos de engorda. Métodos: foram utilizados 48 suínos híbridos (11,74±1,72 kg de peso vivo inicial). Os tratamentos consistiram em quatro níveis (0,00, 0,05, 0,10, 0,15% mais) e dois tipos (regular e protegida) de Met em dietas para suínos de engorda. Resultados: suínos no início, finalização I e II alimentados com Met protegida aumentaram o consumo diário de ração (DFI; p≤0,10). Met protegido aumentou o ganho de peso diário (DWG) em suínos na iniciação e, aumentou espessura da gordura dorsal (BT) em suínos em iniciação e crescimento (p≤0,10). Nos suínos finalização I, Met protegido aumentou DWG e melhorou as características do canal (p≤0,10). Para suínos de iniciação e crescimento, 0,15% extra de Met diminuiu conversão alimentar (FGR; p≤0,10). No crescimento e finalização II, a adição de 0,05% de Met melhoraram o DWG e com 0,10% reduziram a concentração de uréia (p≤0,10). Conclusões: o uso de Met protegida melhora DWG, DFI e BT. O nível de Met aumentado de 0,05-0,15% melhora FGR e DWG; além disso, a excreção de nitrogênio ambiental pode ser diminuída.

Multi-objective optimization of methionine+cysteine levels in diets for growing pigs / Nivel óptimo multiobjetivo de metionina+cisteína en dietas para cerdos en crecimiento / Otimização multiobjetivo do nível de metionina+cisteína em dietas para suínos em crescimento

Abstract Background: The addition of synthetic methionine to growing pig diets should consider the requirements of the methionine+cysteine complex, as most cysteine is obtained through dietary methionine. Objective: To determine an optimal level of methionine+cysteine (Met+Cys) in growing pig diets. Methods: Ninety-nine hybrid (Yorkshire×Landrace×Duroc) barrows were used in a completely randomized design (initial body weight: 25.90±3.99 kg). Nine levels of Met+Cys (0.500, 0.525, 0.550, 0.575, 0.600, 0.625, 0.650, 0.675, and 0.700%) were evaluated during 35 days. To determine a multi-objective optimal level of Met+Cys, growth performance and strongly related carcass traits were evaluated. Results: Average daily feed intake (ADFI) and feed:gain ratio (FGR) showed a linear response to the dietary Met+Cys level (p<0.05), with estimated optimal concentration at 0.700%. Average daily gain (ADG) did not show any effect on the Met+Cys level (p>0.05). Carcass characteristics were not affected by the Met+Cys concentration (p>0.05). The multi-objective optimal level for ADG, ADFI, and FGR was 0.667% Met+Cys. Conclusions: According to our results, 0.667% Met+Cys is the best concentration in diets for growing pigs.

Resumen Antecedentes: Las dietas para cerdos en crecimiento deben adicionarse con metionina sintética para suplir los requerimientos del complejo metionina+cisteína (Met+Cis), pues ellos producen gran parte de la cisteína que requieren a partir de la metionina dietaría. Objetivo: determinar el nivel óptimo multiobjetivo de Met+Cis en dietas para cerdos en crecimiento. Métodos: noventa y nueve machos castrados híbridos (Yorkshire×Landrace×Duroc) se utilizaron en un diseño completamente al azar (peso vivo inicial: 25,90±3,99 kg). Se evaluaron nueve niveles de Met+Cys (0,500, 0,525, 0,550, 0,575, 0,600, 0,625, 0,650, 0,675 y 0,700%) durante 35 d. Para determinar el nivel óptimo multiobjetivo de Met+Cis se consideraron las variables de rendimiento de crecimiento y características de la canal que mantienen una fuerte interrelación. Resultados: el promedio de consumo de alimento diario (ADFI) y la conversión alimenticia (FGR) mostraron una respuesta lineal al nivel de Met+Cis en la dieta (p<0,05), con una concentración óptima estimada en 0,700%. El nivel de Met+Cis no afecto a la ganancia diaria de peso (ADG; p>0,05). La concentración de Met+Cis tampoco afectó las características de la canal (p>0,05). El nivel óptimo multi- objetivo de Met+Cis para ADFI, FGR y ADG fue 0,667%. Conclusión: los resultados indican que 0,667% de Met+Cis es la mejor concentración en la dieta para cerdos en crecimiento.

Resumo Antecedentes: a adição de metionina em dietas para suínos em crescimento deve ser feita para atender às exigências do complexo metionina+cisteína (Met+Cis), porque grande parte da cisteína é obtida através da dieta com metionina. Objetivo: determinar o nível multiobjetivo ótimo de Met+Cis em dietas para suínos em crescimento. Métodos: noventa e nove suínos machos castrados híbridos (Yorkshire×Landrace×Duroc) foram utilizados em delineamento inteiramente casualizado, com peso inicial de 25,90±3,99 kg. Nove níveis de Met+Cis (0,500, 0,525, 0,550, 0,575, 0,600, 0,625, 0,650, 0,675 y 0,700%) foram avaliados em suínos em crescimento por 35 d. Para determinar o nível multiobjetivo ótimo de Met+Cis, foram consideradas as variáveis de comportamento de crescimento e características do canal que mantêm uma forte inter-relação. Resultados: a ingestão média diária de alimentos (ADFI) e a conversão alimentar (FGR) mostraram uma resposta linear ao nível de Met+Cis na dieta (p<0,05), estimando a concentração ótima em 0,700%. O ganho de peso diário (ADG) não mostrou efeito devido ao nível de Met+Cis na dieta (p>0,05). As características da carcaça não foram afetadas pela concentração de Met+Cis na dieta (p>0,05). Com ADFI, FGR e ADG, o nível ótimo de Met+Cis foi obtido, encontrando este nível em 0,667%. Conclusões: os resultados indicam que 0,667% de Met+Cis é a melhor concentração na dieta para suínos em crescimento.

Methionine synthase A2756G polymorphism associated with an increased risk of head and neck cancer

Background:The 5-methyltetrahydrofolate-homocysteine methyltransferase gene (MTR) encodes the methionine synthase enzyme (OMIM 156570). Methionine synthase synthesizes methionine by re-methylation of homocysteine. A single nucleotide variation MTR-A2756G may affects the function of methionine synthase enzyme, which could lead to the development of head and neck squamous cell carcinoma (HNSCC).Methods:In current study, 292 HNSCC patients and 324 normal individuals without any history of cancer (control) were enrolled. EDTA whole blood samples of patients and control individuals were collected, and DNA was extracted. All samples were genotyped for MTR-A2756G polymorphism using polymerase chain reaction-restriction fragment length polymorphism. Frequency of polymorphism was compared between HNSCC patients and control individuals. The association of MTR-A2756G polymorphism with risk factors was statistically analysed through multivariate analysis (multiple logistic regression) whereas univariate analysis (chi square) was performed for group comparisons.Results:Univariate analysis revealed that the frequency of groups like age, smoking and MTR-A2756G genotype was different in HNC patients and controls (p value <0.05). Multivariate analysis showed that smoking (adjusted OR, 3.7; 95% CI, 2.3-6.0), age groups 41-50 years (adjusted OR, 3.6; 95% CI, .9-6.7) and >60 years (adjusted OR, 3.5; 95% CI, 1.7-7.3), MTR-A2756G genotype (adjusted OR, 2.1; 95% CI, 1.3-3.5) is associated with increased risk of HNSCC.Conclusions:Our data suggests that the MTR-A2756G polymorphism is associated with the occurrence of HNSCC in Pakistani population while the individuals between 40 to 50 years of age and tobacco smokers are at a greater risk of developing HNSCC.

Effect of Dietary Supplementation of Critical Amino Acids and Multi-Enzyme with Low Protein and Energy Diet on Carcass and Blood Biochemical Performance of Broiler Chicken

The experiment was conducted to study the effect of dietary supplementation of critical amino acids and multi-enzyme with reduced levels of energy and protein diet on carcass and blood biochemical performance in broiler chicken. Three hundred broiler chicks were allotted to five dietary treatments of three replications and 20 birds in each replication. The control (T0) were fed with standard diet, T1 (5% low CP and energy than standard with balancing of lysine and methionine), T2 (T1 + Multi-enzyme), T3 (10% low CP and energy than standard with balancing of lysine and methionine), T4 (T3 + Multi-enzyme).The dressing percentage and edible meat percentage were significantly more in T2 group. The breast meat percentage of the control group was comparable with T2 group. The result showed that the significantly higher (P<0.05) gizzard weights in groups receiving enzyme combination supplementation as compared to control group (T0) irrespective of the energy-protein levels. The mean value of total protein, albumin, globulin and blood urea nitrogen of all the experimental birds were found to be statistically similar with that of the control group. The treatment group T2 found to be effective and economical

Protective effect of Xiaochaihu Decoction on non-alcoholic steatohepatitis model mice / 中草药

Objective: The protective effect of Xiaochaihu Decoction on non-alcoholic steatohepatitis (NASH) model mice was studied by constructing a methionine-choline deficiency (MCD) diet-induced NASH model in mice. Methods: C57BL/6 mice, as the research objects, were randomly divided into normal control group, model group, Xiaochaihu Decoction (high, medium and low doses) group, Yishanfu group and Qianggan Capsule group. The NASH model was established by feeding MCD feeds, and model intervention was carried out at the same time by giving different drugs in groups; Changes in body weight, daily food intake, and daily water volume of the mice were recorded during the experiment. HE staining of liver tissue was performed at the end of the experiment to observe pathological changes. and the levels of biochemical indicator of alanine aminotransferase (ALT), Aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) in serum, and the changes of TC and TG levels in liver tissue were detected, RT-PCR was used to detect the expression levels of fatty acid synthase (FAS) and sterol regulatory element binding protein 1c (SREBP-1c) in liver tissues. Results: Data such as mouse weight, daily food intake, daily water intake, and organ coefficients indicated that MCD diet-induced model mice showed weight loss, decreased intake, and decreased liver wet weight. The weight, intake and liver coefficient of mice in Xiaochaihu Decoction group were significantly higher than those in the model group; The results of HE staining showed that Xiaochaihu Decoction could significantly reduce the degree of steatosis and inflammation of liver tissue, and improve the morphology and structure of liver cells; The results of serum biochemical indicators showed that Xiaochaihu Decoction significantly reduced the levels of TG, TC, AST, ALT, IL-6, TNF-α and increased the level of HDL-C in NASH model mice; RT-PCR results showed that the gene expression levels of FAS and SREBP-1c in the liver tissue of the model group mice were significantly increased, and the administration of Xiaochaihu Decoction could significantly reduce the gene expression levels of FAS and SREBP-1c. Conclusion: Xiaochaihu Decoction has obvious protective effect on NASH mouse model induced by MCD diet. It may play a lipid-lowering role by regulating the expression of inhibitors of fatty acid synthetase genes (FAS, SREBP-1c), reducing fat accumulation, and inhibiting the expression of inflammatory factors to improve liver tissue damage.

Analysis on Urine Metabolomics of Dioscoreae Nipponicae Rhizoma Extract Against Acute Gouty Arthritis / 中国实验方剂学杂志

Objective::To explore the effect of Dioscoreae Nipponicae Rhizoma extract (DNRe) on rats with acute gouty arthritis (AGA) based on urine metabolomics and to search for the related potential biomarkers and metabolic pathways. Method::Rat model of AGA induced by monosodium urate (MSU) was selected, 40 rats were randomly divided into the blank group (k), the DNRe group (g), the model group (m), and the DNRe treatment group (gm), with 10 rats in each group. The drug-administered group was administered with DNRe at a dose of 0.48 g·kg-1 once a day for 5 days. The urine was gathered after the last administration, and analyzed with UPLC-Q-TOF/MS coupled with pattern recognition techniques, electrospray ionization (ESI) under positive and negative ion scanning mode was adopted, data collection range was m/z 100-1 500 with full scanning mode. Result::A total of 12 common potential biomarkers were identified as sarcosine, dimethylglycine, deoxycytidine, uric acid, 5-hydroxytryptamine (5-HT), L-cystathionine, 4-pyridoxic acid, deoxyuridine, melatonin, 5-methoxytryptamine, fumaric acid and cytidine. Compared with the blank group, the 12 potential biomarkers in the DNRe group were significantly down-regulated. Compare with the model group, 10 metabolites were up-regulated and 2 metabolites were down-regulated in the 12 potential biomarkers of the DNRe treatment group, the abnormal expression of 10 markers including sarcosine, uric acid, L-cystathionine, 4-pyridoxic acid, deoxyuridine, 5-methoxytryptamine, cytidine, dimethylglycine, melatonin, fumaric acid could be modulated by DNRe. The strongest metabolic pathways associated with AGA were cysteine and methionine metabolism, and tryptophan metabolism. Conclusion::The effect of DNRe on AGA may be related to the promotion of conversion level from cystathionine to cysteine in the cysteine and methionine metabolism, and the up-regulating melatonin level in tryptophan metabolism.