High Levels Of Methionine Research Articles

Proximate Analysis of the Paste of Three Varieties of Peanuts (Arachis Hypogaea) Consumed in Côte D’ivoire

Peanuts (Arachis hypogaea) since colonization have been an obligatory crop in Ivory Coast. It is currently the culture of women par excellence in the North of Côte d'Ivoire and especially in Korhogo. Almost the entire production is consumed on site and mainly in the form of peanut paste. The objective of this study is to determine the nutritional composition of peanut pastes of three varieties produced in Côte d'Ivoire. To achieve this, three varieties of peanut were studied and named according to their morphological characteristics (large striated shell (Gcs), medium striated shell (Mcs) and small smooth shell (Pcl). These grains were transformed into paste then subjected to analyzes to know their physicochemical characteristics. The results of statistical analysis showed significant differences at P  0.05. The variety that contains the least water is (Pcl), it will better resist microbial proliferation. Gcs peanut paste was much richer in fat with a value of 45.088  0.19% and also contained myristic acid (2.976  0.008%) and linolenic acid (0.263  0.002%). As for the Pcl variety, the linoleic and oleic acid contents were dominant compared to the other varieties respectively with values of 1.139  0.006 and 0.412  0.002%. Furthermore, regarding proteins, the Pcl variety presented a high level of proteins (29.00  1.75%), however, arginine, methionine, cysteine and glutamic acid are absent.

Association of Long-term Exposure to Air Pollution and Dementia Risk: The Role of Homocysteine, Methionine, and Cardiovascular Burden.

Growing evidence links air pollution with dementia risk, but the biological mechanisms are largely unknown. We investigated the role played by homocysteine (tHcy) and methionine in this association and explored whether this could be explained by cardiovascular diseases (CVDs). Data were extracted from the ongoing Swedish National study on Aging and Care in Kungsholmen (SNAC-K), a longitudinal population-based study. At baseline, 2,512 dementia-free participants were examined up to 2013 (mean follow-up: 5.18 ± 2.96 years). Two air pollutants (particulate matter ≤2.5 μm [PM2.5] and nitrogen oxides [NOx]) were assessed yearly from 1990 until 2013 using dispersion models at residential addresses. The hazard ratio of dementia over air pollution levels was estimated using Cox models adjusted for age, sex, education, smoking, socioeconomic status, physical activity, retirement age, creatinine, year of assessment, and the use of supplements. The total effect of air pollutants on dementia was decomposed into 4 pathways involving tHcy/methionine: (1) direct effect; (2) indirect effect (mediation); (3) effect due to interaction; and (4) effect due to both mediation and interaction. To test whether the association was independent from CVDs (ischemic heart disease, atrial fibrillation, heart failure, and stroke), we repeated the analyses excluding those individuals who developed CVDs. The mean age of the study participants was 73.4 years (SD: 10.4), and 62.1% were female individuals. During an average period of 5 years (mean: 5.18; SD: 2.96 years), 376 cases with incident dementia were identified. There was a 70% increased hazard of dementia per unit increase of PM2.5 during the 5 years before baseline (hazard ratio [HR]: 1.71; 95% CI 1.33-2.09). Overall, 50% (51.6%; 95% CI 9.0-94.1) of the total effect of PM2.5 on dementia was due to mediation of tHcy (6.6%; 95% CI 1.6-11.6) and/or interaction (47.8%; 95% CI 4.9-91.7) with tHcy and 48.4% (p = 0.03) to the direct effect of PM2.5 on dementia. High levels of methionine reduced the dementia hazard linked to PM2.5 by 31% (HR: 0.69; 95% CI 0.56-0.85) with 24.8% attributable to the interaction with methionine and 25.9% (p = 0.001) to the direct effect of PM2.5. No mediation effect was found through methionine. Attenuated results were obtained for NOx. Findings for tHcy were attenuated after excluding those who developed CVDs, while remained similar for methionine. High levels of homocysteine enhanced the dementia risk attributed to air pollution, while high methionine concentrations reduced this risk. The impact of homocysteine on cardiovascular conditions partly explains this association. Alternative pathways other than cardiovascular mechanisms may be at play between methionine and dementia.

Reactions of Finger Millet (Eleusine coracana (L.) Gaertn) Genotypes for Blast Resistance and Yield in Wukari and Takum Taraba State, Nigeria

Finger millet contains high level of methionine, tryptophan, vitamin B, fibre and minerals such as phosphorus, iron, calcium, which serve a good source of balance diet formulations for hypertension, obesity, diabetic patients, pregnant women, nursing mothers, children, people leaving with HIV, malnourished people, incidence of iron deficiency (anemia), and calcium deficiency causing rickets in young children. Production constraints responsible for the low yields have been identified as pests, blast and Striga, drought, low soil fertility, labor intensity, high weed infestation, low yielding varieties, lodging, and poor attitude to the crop. Finger millet blast about 50% yield losses. This disease has been identified as the highest priority constraint to finger millet production in Nigeria, since most landraces and a number of other genotypes are highly susceptible. Field trials were conducted on 22 germplasm (U 15, P224, ACC 32, ACC14, KNE814, IE 3779, KNE 628, KNE 688, KNE 1149, Etiyo Brown, Gulu E, Kala, RW 127(IE 6613), KNE 392, GBK 029681, ACC 3953, GBK 011136A, KNE 689, KNE 1034, Emiroit, Red local check and Black local check) from ICRISAT and farmers in 3 replicates at Wukari and Takum in 2019, to assess for blast disease resistance and yield. A significant negative correlation (0.52) was found between blast severity and DF (days to flowering) suggesting that late/medium flowering varieties are more resistant than the early ones as indicated by genotypes KNE814, IE 3779, KNE 628, KNE 688, KNE 1149, Gulu E,, KNE 392, GBK 029681, ACC 3953, GBK 011136A, KNE 689, KNE 1034, Emiroit and Red (local check) varieties, which were medium maturing and highly resistance to blast disease, with foliar blast severity (1.07 - 3.0), neck blast (1.5 - 2.5) and finger blast (11 - 25 %), NET (3.45 - 4.38), NFPH (5.80) and subsequently highest yield (1565 - 1887 kg/ha), compared to black local variety with the highest foliar blast severity (4.00 - 6.70), neck blast (2.7 - 4.40), finger blast (32 - 36 %), NET (2.50 - 2.70), NFPH (3.30 - 4.70) and lowest yield (688 - 989 kg/ha), respectively in Wukari and Takum locations. Hence, these varieties resistant to foliar, neck, finger blast diseases, good yield parameters are recommended as sources of germplasm for genetic improvement and further research.

Reactions of Finger Millet (Eleusine coracana (L.) Gaertn) Genotypes for Blast Resistance and Yield in Wukari and Takum Taraba State, Nigeria

Finger millet contains high level of methionine, tryptophan, vitamin B, fibre and minerals such as phosphorus, iron, calcium, which serve a good source of balance diet formulations for hypertension, obesity, diabetic patients, pregnant women, nursing mothers, children, people leaving with HIV, malnourished people, incidence of iron deficiency (anemia), and calcium deficiency causing rickets in young children. Production constraints responsible for the low yields have been identified as pests, blast and Striga, drought, low soil fertility, labor intensity, high weed infestation, low yielding varieties, lodging, and poor attitude to the crop. Finger millet blast about 50% yield losses. This disease has been identified as the highest priority constraint to finger millet production in Nigeria, since most landraces and a number of other genotypes are highly susceptible. Field trials were conducted on 22 germplasm (U 15, P224, ACC 32, ACC14, KNE814, IE 3779, KNE 628, KNE 688, KNE 1149, Etiyo Brown, Gulu E, Kala, RW 127(IE 6613), KNE 392, GBK 029681, ACC 3953, GBK 011136A, KNE 689, KNE 1034, Emiroit, Red local check and Black local check) from ICRISAT and farmers in 3 replicates at Wukari and Takum in 2019, to assess for blast disease resistance and yield. A significant negative correlation (0.52) was found between blast severity and DF (days to flowering) suggesting that late/medium flowering varieties are more resistant than the early ones as indicated by genotypes KNE814, IE 3779, KNE 628, KNE 688, KNE 1149, Gulu E,, KNE 392, GBK 029681, ACC 3953, GBK 011136A, KNE 689, KNE 1034, Emiroit and Red (local check) varieties, which were medium maturing and highly resistance to blast disease, with foliar blast severity (1.07 - 3.0), neck blast (1.5 - 2.5) and finger blast (11 - 25 %), NET (3.45 - 4.38), NFPH (5.80) and subsequently highest yield (1565 - 1887 kg/ha), compared to black local variety with the highest foliar blast severity (4.00 - 6.70), neck blast (2.7 - 4.40), finger blast (32 - 36 %), NET (2.50 - 2.70), NFPH (3.30 - 4.70) and lowest yield (688 - 989 kg/ha), respectively in Wukari and Takum locations. Hence, these varieties resistant to foliar, neck, finger blast diseases, good yield parameters are recommended as sources of germplasm for genetic improvement and further research.

Effect of Increasing the Methionine Level and Reducing the Threonine Level in the Diet of Fast-Growing Rabbits.

The main aim of this work was to evaluate a novel combination of the first limiting amino acids (lysine, methionine, and threonine) in fast-growing rabbits (combination MHL, shown to minimise levels of plasmatic urea nitrogen in previous research, medium level of lysine, high level of methionine, and low level of threonine) and compare it with current recommendations (combination MMM, medium level of all amino acids). A total of 165 weaned rabbits (28 d old) from a paternal line selected for growth rate were used in the growth trial. The effect of the diet on apparent fecal digestibility, as well as the apparent and true ileal digestibility, were studied. Nutrient retention was also determined. Although no differences in digestibility between diets were observed, animals fed with Diet MHL improved the global average daily gain (+2.3 g/d; p = 0.0482) and feed conversion ratio (-0.10; p = 0.0229). Animals fed with Diet MMM reduced the protein-to-energy ratio retained (p = 0.0086). In conclusion, Diet MHL promoted an improvement in growth traits in a paternal line. Consequently, we propose its levels of 6.4, 5.4, and 5.0 g/kg DM of true ileal digestible lysine, methionine, and threonine, respectively.

Dietary Methionine Level Impacts the Growth, Nutrient Metabolism, Antioxidant Capacity and Immunity of the Chinese Mitten Crab (Eriocheir sinensis) under Chronic Heat Stress.

This study examined whether diets with high dietary methionine levels could alleviate chronic heat stress in Chinese mitten crab Eriocheir sinensis. Crabs were fed three dietary methionine levels of 0.49%, 1.29% and 2.09% for six weeks. The analyzed methionine concentration of diets was 0.48%, 1.05% and 1.72%, respectively. Crabs were fed three different supplemental concentrations of dietary methionine at 24 °C and 30 °C, respectively. The trial was divided into six groups with five replicates in each group, and 40 juvenile crabs (initial average weight 0.71 ± 0.01 g) in each replicate. During the trial, crabs were fed twice daily (the diet of 4% of the body weight was delivered daily). The effects of dietary methionine level on nutrient metabolism, antioxidant capacity, apoptosis factors and immunity were evaluated at a normal water temperature of 24 °C and high temperature of 30 °C. Feed conversion ratio decreased under chronic heat stress. Chronic heat stress increased weight gain, specific growth rate, molting frequency, and protein efficiency ratio. The survival of crabs decreased under chronic heat stress, whereas a high level of dietary methionine significantly improved survival. Chronic heat stress induced lipid accumulation and protein content reduction. The high-methionine diet decreased lipid in the body and hepatopancreas, but increased protein in the body, muscle and hepatopancreas under chronic heat stress. Simultaneously, the high dietary methionine levels mitigated oxidative stress by reducing lipid peroxidation, restoring the antioxidant enzyme system, decreasing apoptosis and activating immune function under chronic heat stress. This study suggests that supplementing 1.72% dietary methionine could alleviate the adverse effects of a high water temperature in E. sinensis farming.

Metoclopramide as a Potential Antipsychotic Against Long-Term Methionine Exposure in Zebrafish.

Methionine (MET) contributes to brain function and is required for proper functioning of the central nervous system. However, exceptionally high levels of MET and its metabolites in plasma have been found to be toxic and can lead to cell alterations. Long-term exposure to MET has been shown to mimic psychotic symptoms in schizophrenic patients and rodents. The present study evaluated behavioral and neurochemical effects of long-term exposure to MET in zebrafish. Five groups of zebrafish were exposed to MET at a concentration of 4.5 mM for 7 days, along with acute exposure to 25 μM of clozapine and 750, 1000, and 1250 μM of metoclopramide. In contrast, the normal group was exposed to only water and dimethyl sulfoxide. After the treatment, social interaction, anxiety, memory, and locomotion of zebrafish and serotonin levels in zebrafish brains were evaluated. Our results showed that metoclopramide was not only beneficial in improving MET-induced cognitive impairment but it also prevented social withdrawal in zebrafish exposed to MET. In addition, metoclopramide reversed anxiety-like behavior, as indicated by significant changes in locomotion activity. Despite slight changes in serotonin levels in the zebrafish brain, an in vitro serotonin assay failed to demonstrate significant differences between the disease control, normal, and two treatment groups. Finally, results from the study showed that repeated administration of MET induced schizophrenia-like symptoms, although metoclopramide ameliorated the MET-mediated negative symptoms and cognitive deficits in zebrafish. Overall, our findings suggest a new perspective to further explore the antipsychotic properties of metoclopramide.

Transient Methionine Deprivation Triggers Histone Modification and Potentiates Differentiation of Induced Pluripotent Stem Cells.

Human induced pluripotent stem cells (iPSCs) require high levels of methionine (Met). Met deprivation results in a rapid decrease in intracellular S-adenosyl-methionine (SAM), poising human iPSCs for differentiation and leading to the apoptosis of undifferentiated cells. Met deprivation triggers rapid metabolic changes, including SAM, followed by reversible epigenetic modifications. Here, we show that short-term Met deprivation impairs the pluripotency network through epigenetic modification in a 3D suspension culture. The trimethylation of lysine 4 on histone H3 (H3K4me3) was drastically affected compared with other histone modifications. Short-term Met deprivation specifically affects the transcription start site (TSS) region of genes, such as those involved in the transforming growth factor β pathway and cholesterol biosynthetic process, besides key pluripotent genes such as NANOG and POU5F1. The expression levels of these genes decreased, correlating with the loss of H3K4me3 marks. Upon differentiation, Met deprivation triggers the upregulation of various lineage-specific genes, including key definitive endoderm genes, such as GATA6. Upon differentiation, loss of H3K27me3 occurs in many endodermal genes, switching from a bivalent to a monovalent (H3K4me3) state. In conclusion, Met metabolism maintains the pluripotent network with histone marks, and their loss potentiates differentiation.

MTORC1-c-Myc pathway rewires methionine metabolism for HCC progression through suppressing SIRT4 mediated ADP ribosylation of MAT2A

BackgroundExploiting cancer metabolism during nutrient availability holds immense potential for the clinical and therapeutic benefits of hepatocellular carcinoma (HCC) patients. Dietary methionine is a metabolic dependence of cancer development, but how the signal transduction integrates methionine status to achieve the physiological demand of cancer cells remains unknown.MethodsLow or high levels of dietary methionine was fed to mouse models with patient-derived xenograft or diethyl-nitrosamine induced liver cancer. RNA sequence and metabolomics were performed to reveal the profound effect of methionine restriction on gene expression and metabolite changes. Immunostaining, sphere formation assays, in vivo tumourigenicity, migration and self-renewal ability were conducted to demonstrate the efficacy of methionine restriction and sorafenib.ResultsWe discovered that mTORC1-c-Myc-SIRT4 axis was abnormally regulated in a methionine-dependent manner and affected the HCC progression. c-Myc rewires methionine metabolism through TRIM32 mediated degradation of SIRT4, which regulates MAT2A activity by ADP-ribosylation on amino acid residue glutamic acid 111. MAT2A is a key enzyme to generate S-adenosylmethionine (SAM). Loss of SIRT4 activates MAT2A, thereby increasing SAM level and dynamically regulating gene expression, which triggers the high proliferation rate of tumour cells. SIRT4 exerts its tumour suppressive function with targeted therapy (sorafenib) by affecting methionine, redox and nucleotide metabolism.ConclusionsThese findings establish a novel characterization of the signaling transduction and the metabolic consequences of dietary methionine restriction in malignant liver tissue of mice. mTORC1, c-Myc, SIRT4 and ADP ribosylation site of MAT2A are promising clinical and therapeutic targets for the HCC treatment.

Chemical hypermethioninemia in young mice: oxidative damage and reduction of antioxidant enzyme activity in brain, kidney, and liver.

High levels of methionine (Met) and its metabolites, such as methionine sulfoxide (MetO), found in hypermethioninemia, can be detrimental to the body; however, the underlying mechanisms are still uncertain. Using a recently standardized protocol, the aim of this study was to investigate the effects of chronic administration of Met and/or MetO on parameters of oxidative damage in the total brain, liver, and kidney of young mice. Swiss male mice were subcutaneously injected with Met and MetO at concentrations of 0.35-1.2g/kg body weight and 0.09-0.3g/kg body weight, respectively, from the 10th-38th day post-birth, while the control group was treated with saline solution. Results showed that Met and/or MetO caused an increase in reactive oxygen species (ROS) and lipoperoxidation, along with a reduction of superoxide dismutase (SOD) and catalase (CAT) activities in the brain. In the liver, Met and/or MetO enhanced ROS and nitrite levels, and reduced SOD, CAT, and delta aminolevulinic dehydratase activities. The effects on the kidney were an increase in ROS production and SOD activity, and a reduction in thiol content and CAT activity. These data demonstrated the contribution of redox imbalance to the systemic changes found in patients with hypermethioninemia. In conclusion, our findings may help future studies to better understand the pathophysiological mechanisms of hypermethioninemia as well as contribute to the search for new therapeutic agents for this pathology.

Compensatory Mechanism of Diet Containing Sulphur-Rich Amino Acids in Restoring Neurotoxico-Nutritional Deficits in Konzo Disease Rat Model

Introduction: The present study is aimed at assessing the effect of bitter cassava on blood biochemical parameters of konzo disease induced Wistar rats. Nutritional modification with suphur rich amino acids was used as a measure to correct the impact in rats induced konzo disease. Method: 25 adult male wistar rats were assigned to 5 experimental groups (i) Control n=5, (ii) cassava only n=5, (iii) cassava + animal feed n=5, (iv) cassava + Eggshell + Brown beans n=5, (v) Eggshell + Brown Beans only n=5. The bitter cassava foods were taken by oral ingestion for a period of 4 weeks. The weights of the rats were checked through the experiment and blood samples were taken from each group into EDTA-lined containers and analyses for blood cyanide, methionine and cysteine levels.
 Results: There was significant difference in weight and there was a progressive increase in their success rate as against the cassava only group which decreased in their success rate, hence, differs statistically and significantly (p˂0.05) from the control group. It was observed that the cassava fed group had higher blood level of cyanide far above the normal blood reference range (2.60 – 2.90µg/ml) for cyanides, hence, was seen to be statistically significant as compared to that of the control group. The eggshell and brown beans only group showed high blood levels of methionine that statistically differ significantly (p<0.05) from both the control group and the cassava only group. Blood level of cysteine in the cassava plus eggshell and brown beans group differed significant statistically from the control group.
 Conclusion: Sulphur amino acids such as methionine and cysteine are essential for detoxification of the residual cyanogens remaining in insufficiently processed cassava roots. Foods such as cereals and legumes as source of sulphur amino acids should be promoted to prevent paralytic neurotoxico-nutritional disease such as konzo among the poor population.

Low protein with high amino acid diets improves the growth performance of yellow feather broilers by improving intestinal health under cyclic heat stress

This study investigated the effects of a low-protein diet supplemented with high lysine, methionine, and threonine levels on production performance, antioxidant capacity, intestinal morphology, duodenum gene expression and intestinal microorganisms in yellow feather broilers under cyclic heat stress. A total of 162 yellow feather broilers that were 42 d of age were selected and randomly divided into three groups: the control group (CONT, CP 16%), heat stress group (HS, CP 16%), and heat stress with low protein and high amino acid group (HS_LP, CP 14.5%). Following 14 d heating stress period, the HS_LP group showed no significant effect on production performance compared with the HS group. After a 28 d feeding trial, compared with the HS group, the HS_LP group significantly reduced feed: gain at 15–28 d (P < 0.05), had a tendency to reduce feed: gain at 1–28 d (P = 0.056). Compared with the HS group, the serum catalase enzyme activity tended to be higher (P = 0.067), and liver glutathione peroxidase enzyme activity was significantly decreased (P < 0.05) in the HS_LP group. Other antioxidant capacity indexes were not significantly different between the two groups (P > 0.05). Duodenum (P < 0.05) and ileum (P < 0.05) villus height were significantly increased, duodenum villus height: crypt depth ratio (P < 0.05) and jejunum crypt depth (P < 0.05) were significantly decreased, and jejunum villus height was significantly reduced (P < 0.05) in broilers fed the HS_LP diet. Compared with the HS group, the mRNA level of Claudin-1 was significantly increased (P < 0.05), and had a tendency to increase Occludin (P = 0.060) in the HS_LP group. In addition, the HS_LP group significantly increased Nitrosomonas abundance (P < 0.05) and had a tendency to reduce unidentified_Mollicutes abundance (P = 0.083) at the genus level compared with the HS group. This information is useful to formulate diets that correct the decrease in amino acid consumption associated with the reduced voluntary feed intake of broilers under heat stress.

Proximate composition and amino acid profile of European eel skin: influence of body weight

Fish processing industries worldwide are discarding million kilograms of fish waste per year. The disposal of these wastes is an important handicap in fish processing industry. The thick eel skin is considered a waste product in the fish processing industry and in the home and is usually converted into low market-value products. The aims of this study were to evaluate the variations in the general chemical composition and amino acid content of eel skin in relation to body weight. The moisture content decreased and the lipid content increased with the weight of the eels. Eel skin had a high protein content, which did not vary significantly with eel weight. Glycine was the most abundant amino acid. The predominant essential amino acid was leucine. The smaller eels contained higher levels of threonine, valine, isoleucine, leucine, lysine, glutamic acid and tyrosine than the other groups. The larger eels contained higher levels of methionine, hydroxyproline, glycine, arginine, alanine and proline than the other groups. The protein indices showed that the skin from smaller eels was of higher nutritional quality.Graphical abstract

Longitudinal Serum Metabolomics in Extremely Premature Infants: Relationships With Gestational Age, Nutrition, and Morbidities.

An increasing number of extremely premature infants survive the neonatal period and beyond. Little is known about the maturation of the preterm infant’s metabolome and its relation to the development of morbidities. Using 1H-NMR, we investigated the serum metabolic profile of 87 infants born at a gestational age (GA) <28 weeks [mean GA (SD) 25.4 (1.4) weeks] in samples longitudinally collected from birth to term equivalent age. The infant metabolome was analyzed in relation to GA, postnatal age, nutrition, and preterm morbidities. At postnatal day 1, low GA correlated with high levels of 3-hydroxyisobutyrate, acetate, acetoacetate, acetone, formate, glucose, and valine. Nearly all quantified metabolites displayed postnatal concentration changes. For example, the two phospholipid-related metabolites myo-inositol and ethanolamine displayed a similar decline from birth over the first weeks of life, irrespectively of GA. The proportion of enteral/parenteral energy intake in the first 28 days significantly correlated with mean levels of 52% of the analyzed metabolites. Low enteral energy intake was associated with high serum levels of 3-hydroxyisobutyrate, creatinine, glucose, glycerol, histidine, lactate, leucine, lysine, methionine, ornithine, phenylalanine, proline, threonine, and uridine. There were also significant correlations between high enteral intake and high serum levels of isoleucine and tyrosine. Retinopathy of prematurity (ROP) and bronchopulmonary dysplasia (BPD) outcomes were not significantly associated with metabolite levels in the neonatal period after correcting for multiple testing. In conclusion, the serum metabolome of extremely premature infants changes substantially in the neonatal period, largely driven by the gradual transfer from total parenteral nutrition to full enteral feeding. Further studies are needed to disentangle the intricate relationships between the metabolome, nutritional management, GA, and the development of preterm morbidities.

Anthropometric, biochemical, and haematological indicators associated with hyperhomocysteinemia and their relation to global DNA methylation in a young adult population

ABSTRACT Increased homocysteine (Hcy) levels have been associated with a higher risk of cardiovascular and neurodegenerative diseases. Passive DNA demethylation has been suggested as one of the mechanisms implicated in the development of these conditions, and most studies have investigated this relationship in older adult populations. Therefore, this study aimed to evaluate the relationship between corporal composition and biochemical and haematological indicators with plasma homocysteine levels and genome-wide methylation (Alu, LINE-1, and SAT2) in a population of healthy young adults (median age, 18 years). We showed that the prevalence of hyperhomocysteinemia was significantly higher in men (18.5%) than in women (6.6%) (P = 0.034). Increased Hcy level was substantially associated with higher levels of body mass index and visceral fat in females, whereas in males, it was significantly associated with reduced red cell distribution width and high-density lipoprotein (HDL) cholesterol (HDL-C) levels and increased low-density lipoprotein/HDL ratio. Hypomethylation of Alu was significantly associated with reduced levels of HDL-C (<40.0 mg dL–1), whereas hypomethylation of LINE-1 and SAT2 was significantly associated with higher levels of skeletal muscle (<39.3%) in males. These results highlight the participation of hormonal factors in regulating Hcy metabolism, primarily in the female population, whereas changes in DNA methylation observed in males might be associated with the consumption of a protein diet with high levels of methionine, independent of increased Hcy levels.

Identification of three novel pathogenic mutations in cystathionine beta-synthase gene of Pakistani intellectually disabled patients.

Classical homocystinuria (HCU) is an autosomal recessive inborn error of metabolism, which is caused by the cystathionine-β-synthase (CBS: encoded by CBS) deficiency. Symptoms of untreated classical HCU patients include intellectual disability (ID), ectopia lentis and long limbs, along with elevated plasma methionine, and homocysteine. A total of 429 ID patients (age range: 1.6-23 years) were sampled from Northern areas of Punjab, Pakistan. Biochemical and genetic analyses were performed to find classical HCU disease in ID patients. Biochemically, nine patients from seven unrelated families were identified with high levels of plasma methionine and homocysteine. Targeted exonic analysis of CBS confirmed seven causative homozygous mutations; of which three were novel missense mutations (c.451G>T; p.Gly151Trp, c.975G>C; p.Lys325Asn and c.1039+1G>T splicing), and four were recurrent variants (c.451+1G>A; IVS4+1 splicing, c.770C>T; p.Thr257Met, c.808_810del GAG; p.Glu270del and c.752T>C; p.Leu251Pro). Treatment of patients was initiated without further delay with pyridoxine, folic acid, cobalamin, and betaine as well as dietary protein restriction. The immediate impact was noticed in behavioral improvement, decreased irritability, improved black hair color, and socialization. Overall, health outcomes in this disorder depend on the age and symptomatology at the time of treatment initiation. With personalized treatment and care, such patients can reach their full potential of living as healthy a life as possible. This screening study is one of the pioneering initiatives in Pakistan which would help to minimize the burden of such treatable inborn errors of metabolism in the intellectually disabled patients.

COMPARATIVE UTILIZATION OF COOKED SOYBEANS AND FISHMEAL-GROUNDNUT CAKE MIXTURE IN DIETS

A 4 week experiment involving 45 growing albino rats was carried out to assess the nutritional utilization of cooked full-fat soybean (CFFS) as a major source of dietary protein relative to that of the protein concentrates fishmeal-groundnut cake mixture and the response to supplementation of the cooked soybeans with 2g DL-methionine/kg diet and/or 2g L-lysine/kg diet. Feed intake (FI), average daily weight gain (DLG),. feed conversion ratio (FCR), protein efficiency ratio (PER) and feed cost (FC) per unit live-weight gain, (FC/WG) were measured, No significant effect of the various dietary treatments was observed on PER and FCIWG. However, the incorporation of CFFS either alone or in conjunction with either of both lysine and methionine significantly improved Fl and FCR (P&lt; 0.05) and also DLG (P&lt; 0.01) over the control fish-groundnut meal ration. Supplementation of CFFS with lysine either alone or along with methionine appeared unwarranted as methionine supplementation alone resulted in similar or slightly better efficiency of protein (0.247) and overall feed (0.596) utilization. From the results obtained, it appears that maize-based CFFS diet supplemented with 2 g/kg DL-methionine may be applicable for lion- ruminant feeding. However, since this level of methionine supplementation only provides for 67% of rat requirement, it is suggested that further studies with higher methionine levels be conducted.

High levels of blood glutamic acid and ornithine in children with intellectual disability

Objectives: Aminoacidopathies are inborn errors of metabolism (IEMs) that cause intellectual disability in children. Luckily, aminoacidopathies are potentially treatable, if diagnosed earlier in life. The focus of this study was the screening of aminoacidopathies in a cohort of patients suspected for IEMs. Methods: Blood samples from healthy (IQ > 90; n = 391) and intellectually disabled (IQ < 70; n = 409) children (suspected for IEMs) were collected from different areas of Northern Punjab, Pakistan. An analytical HPLC assay was used for the screening of plasma amino acids. Results: All the samples (n = 800) were analyzed on HPLC and forty-three out of 409 patient samples showed abnormal amino acid profiles mainly in the levels of glutamic acid, ornithine and methionine. Plasma concentration (Mean ± SD ng/mL) were significantly high in 40 patients for glutamic acid (patients: 165 ± 38 vs. controls: 57 ± 8, p < 0.00001) and ornithine (patients: 3177 ± 937 vs. controls: 1361 ± 91, p < 0.0001). Moreover, 3 patients showed abnormally high (53.3 ± 8.6 ng/mL) plasma levels of methionine. Conclusion: In conclusion, biochemical analysis of samples from such patients at the metabolites level could reveal the underlying diseases which could be confirmed through advanced biochemical and genetic analyses. Thus, treatment to some of such patients could be offered. Thus burden of intellectual disability caused by such rare metabolic diseases could be reduced from the target populations.

The Level of Methionine Residues in Storage Proteins Is the Main Limiting Factor of Protein-Bound-Methionine Accumulation in Arabidopsis Seeds.

The low level of methionine, an essential sulfur-containing amino acid, limits the nutritional quality of seeds. Two main factors can control the level of protein-bound methionine: the level of free methionine that limits protein accumulation and the methionine residues inside the storage proteins. To reveal the main limiting factor, we generated transgenic Arabidopsis thaliana seed-specific plants expressing the methionine-rich sunflower seed storage (SSA) protein (A1/A2). The contents of protein-bound methionine in the water-soluble protein fraction that includes the SSA in A1/A2 were 5.3- and 10.5-fold, respectively, compared to control, an empty vector (EV). This suggests that free methionine can support this accumulation. To elucidate if the level of free methionine could be increased further in the protein-bound methionine, these lines were crossed with previously characterized plants having higher levels of free methionine in seeds (called SSE). The progenies of the crosses (A1S, A2S) exhibited the highest level of protein-bound methionine, but this level did not differ significantly from A2, suggesting that all the methionine residues of A2 were filled with methionine. It also suggests that the content of methionine residues in the storage proteins is the main limiting factor. The results also proposed that the storage proteins can change their content in response to high levels of free methionine or SSA. This was assumed since the water-soluble protein fraction was highest in A1S/A2S as well as in SSE compared to EV and A1/A2. By using these seeds, we also aimed at gaining more knowledge about the link between high free methionine and the levels of metabolites that usually accumulate during abiotic stresses. This putative connection was derived from a previous analysis of SSE. The results of metabolic profiling showed that the levels of 29 and 20 out of the 56 metabolites were significantly higher in SSE and A1, respectively, that had higher level of free methionine, compared A1S/A2S, which had lower free methionine levels. This suggests a strong link between high free methionine and the accumulation of stress-associated metabolites.

Association of Homocysteine, Methionine, and MTHFR 677C>T Polymorphism With Rate of Cardiovascular Multimorbidity Development in Older Adults in Sweden

Strong evidence links high total serum homocysteine (tHcy) and low methionine (Met) levels with higher risk of ischemic disease, but other cardiovascular (CV) diseases may also be associated with their pleiotropic effects. To investigate the association of serum concentrations of tHcy and Met with the rate of CV multimorbidity development in older adults and to explore the role of methylenetetrahydrofolate reductase (MTHFR) 677C>T polymorphism in this association. The Swedish National Study on Aging and Care in Kungsholmen is a cohort study of randomly selected individuals aged 60 years or older. The present study included data on 1969 individuals with complete information and without CV diseases at baseline, collected from the baseline examination (2001-2004) to the fourth follow-up (2013-2016). Data analysis was conducted from January to May 2019. Concentrations of tHcy and Met were measured from nonfasting venous blood samples. The Met:tHcy ratio was considered a possible indicator of methylation activity. MTHFR status was dichotomized as any T carriers vs noncarriers. The number of CV diseases at each wave was ascertained based on medical interviews and records, laboratory test results, and drug data. Linear mixed models were used to study the association of baseline tHcy and Met levels and the rate of CV multimorbidity development, adjusting for sociodemographic characteristics, CV risk factors, chronic disease burden, and drug use. Of 1969 participants, most were women (1261 [64.0%]), with a mean (SD) age of 70.9 (9.8) years; 1703 participants (86.6%) had at least a high school level of education. Baseline measurements of serum tHcy, Met, and the Met:tHcy ratio were associated with the rate of CV disease accumulation (tHcy: β = 0.023 per year; 95% CI, 0.015 to 0.030; P < .001; Met: β = -0.007 per year; 95% CI, -0.013 to -0.001; P = .02; Met:tHcy ratio: β = -0.017 per year; 95% CI, -0.023 to -0.011; P < .001). The association between low Met concentrations and the rate of CV multimorbidity development was restricted to the group with CT/TT alleles of MTHFR (β = 0.023 per year; 95% CI, 0.006 to 0.041; P = .009). Results remained largely significant when individual CV diseases were removed from the total count 1 at a time (eg, ischemic heart disease, tHcy: β = 0.023 per year; 95% CI, 0.013 to 0.027; P < .001; Met: β = -0.006 per year; 95% CI, -0.011 to -0.0003; P = .04; Met:tHcy ratio: β = -0.015 per year; 95% CI, -0.020 to -0.009; P < .001). In this study, high tHcy and low Met levels were associated with faster CV multimorbidity development in older age. The interactive association of Met concentrations and MTHFR polymorphism, together with the association found for the Met:tHcy ratio, point toward the relevance of impaired methylation in the pathogenesis of CV aging.