Erythrocyte Glutathione Reductase Activity Research Articles

Antioxidant and metabolic adjunctive treatment in late onset psychosis

IntroductionBasing on our previous findings of significant additional gain obtained from usage of adjunctive antioxidant medicine added to antipsychotic+antidepressant therapy in late-onset schizophrenia-like psychoses (LOP), the group often suffering of comorbid pathologies and experiencing substantial side-effects of drugs, we spred our approach to try “metabolic” medicines as adjunctives in LOP.ObjectivesTo reveal biochemical parameters of the blood cells which might be used for distinguishing subgroups of patients suffering with LOP for whom various adjunctive therapy (antioxidant, metabolic) would be advantageous.MethodsThe study included 59 patients 50-89 years old, with LOP (onset after 40 years), and 38 healthy peoples 51 – 84 years old. The activities of glutamate dehydrogenase (GDH), glutathione reductase (GR), and glutathione S-transferase (GST) were determined spectrophotometrically in erythrocytes and platelets. Scores by PANSS were evaluated twice: before and on the 28-th day of antipsychotic treatment.ResultsSamples from control group were used for determination of the control ranges for levels of studied enzymatic activities. Enzymatic activity levels were analyzed in three groups of patients: group Gr1 (n=16) treated without adjunctive therapy, and two other groups (Gr2 and Gr3) treated with adjunctive medicines: antioxidant 2-ethyl-6-methyl-3-hydroxypyridine succinate (Gr2, n=20), or “metabolic” medicines citicoline/cerebrolysin/cortexin/actovegin/gliatilin (Gr3, n=23).As compared with controls, activity of erythrocyte GR was decreased at baseline and after the treatment course in all patients’ groups (p<0.01); in Gr2 significant decreases in baseline platelet GDH and GST activities were observed (p=0.005). Different significant links between biochemical parameters and scores by clinical scales before treatment were observed: in Gr1, erythrocyte GST activity positively correlated with scores by PANSS-Neg (R=0.61, p=0.012), by PANSS-Psy (R=0.54, p=0.032), and by PANSS (R=0.62, p=0.010), in Gr2, erythrocyte GST activity positively correlated with scores by PANSS-Pos (R=0.53, p=0.016), by PANSS-Psy (R=0.52, p=0.015), and by PANSS (R=0.60, p=0.005), in Gr3, platelet GR activity positively correlated with PANSS-Pos (R=0.50, p=0.014).ConclusionsWe have confirmed the additional favor (decrease in side-effect severity) obtained by distinct patient groups when treated with adjunctive antioxidant or “metabolic” therapy. Moreover, correlations revealed in the patient subgroups between enzymatic activities and scores by psychometric scales enable revealing those biochemical markers measurement of which facilitate differentiating the patients for whom the adjunctive medicines to antipsychotic+antioxidant treatment can positively influence the treatment outcome.Disclosure of InterestNone Declared

Late onset psychosis treatment with adjunctive medicines.

A number of studies have shown the feasibility of using adjunctive drugs in late onset psychosis (LOP). Testing hypothesis that among LOP people treated with antipsychotics and antidepressants, basing on certain clinical characteristics a subgroup of patients might be distinguished, for whom adjunctive therapy is advantageous. This subgroup might be identified by measurement of blood biochemical parameters. 59 in-patients with LOP, treated neuroleptics and antidepressants, were included, and followed in real clinical practice. Database containing demographic, clinical data (scores by PANSS, CDSS, CGI-S, HAMD-17), prescribed therapy, adverse effects of antipsychotic and antidepressant treatment, and blood biochemical parameters (enzymatic activities of glutamate- and glutathione metabolism enzymes in platelets and erythrocytes) at baseline and after the treatment course was created. Three groups of patients (Gr1, Gr2, and Gr3), based on the adjunctive therapy usage were identified: Gr1 (n = 16) was without adjunctive therapy, two other groups (Gr2 and Gr3) were with adjunctive medicines, such as 2-ethyl-6-methyl-3-hydroxypyridine succinate (EMHS; Gr2, n = 20), or other drugs, such as citicoline, cerebrolysin, cortexin, actovegin, gliatilin (choline alfoscerate; Gr3, n = 23). The enzymatic activities were assessed also in the matched control group (n = 38). In all three patient groups, as compared with controls, activity of erythrocyte glutathione reductase was decreased at baseline and after the treatment course. In Gr2, unlike Gr1 or Gr3, there was a significant decrease in baseline glutamate dehydrogenase and glutathione-S-transferase activities. Certain clinical criteria were also elucidated for prescription of EMHS as adjunctive therapy for patients of Gr2. Glutamate dehydrogenase and glutathione-S-transferase activities returned closer to control levels after the treatment course in Gr2, unlike Gr1, where they declined yet more after psychotropic treatment without adjunctive medicine. Different significant links between biochemical parameters and scores by clinical scales were observed in Gr1, Gr2, and Gr3, some having predictive value for evaluation of antipsychotic treatment efficacy. We demonstrate the validity of adjunctive neuroprotective medicines' usage in addition to antipsychotic and antidepressant therapy in distinct subgroups of patients suffering with LOP, especially those who have prominent side effects accompanying their psychotropic treatment. Returning of biochemical parameters to control range following the treatment course observed in patients of the subgroup treated with adjunctive EMHS is evidence for their metabolism normalization.

Risk factors for anaemia among women and their young children hospitalised with suspected thiamine deficiency in northern Lao PDR.

Anaemia among women and young children remains a major public health concern. This secondary study describes the anaemia prevalence among young hospitalised children and their mothers in northern Lao People's Democratic Republic and explores possible nutritional causes and risk factors for anaemia. Hospitalised children (ages 21 days to <18 months) with clinical symptoms suggestive of thiamine deficiency disorders were eligible along with their mothers. Venous blood was collected for determination of haemoglobin, ferritin, soluble transferrin receptor (sTfR), retinol-binding protein (RBP), erythrocyte glutathione reductase activation coefficient (EGRac), thiamine diphosphate (ThDP) and acute phase proteins. Risk factors for anaemia were modelled using minimally adjusted logistic regression controlling for age. Haemoglobin results were available for 436 women (mean ± SD age 24.7 ± 6.4 years; 1.6% pregnant) and 427 children (4.3 ± 3.5 months; 60.3% male). Anaemia prevalence (Hb < 120 g/L for nonpregnant women and <110 g/L for pregnant women and children) was 30.7% among women and 55.2% among children. In bivariate analyses, biomarkers significantly associated with anaemia in women were ferritin, sTfR, RBP, EGRacand ThDP. Other risk factors for women were lower BMI, mid-upper arm circumference < 23.5 cm, lower education, lower socioeconomic index, food insecurity, Hmong ethnicity, not/rarely having attended antenatal care, not having taken antenatal iron-containing supplements and not meeting minimum dietary diversity. Risk factors for anaemia among children were older age, male sex, stunting, sTfR, ThDPand alpha-1-acid-glycoprotein. Anaemia was common among women and their hospitalised children and was associated with micronutrient deficiencies and socioeconomic,dietary and health care-seeking risk factors, suggesting that multiple strategies are required to prevent anaemia among women and children.

Effect of Cadmium on Oxidative Stress Indices and Vitamin D Concentrations in Children

Heavy metal poisoning can have serious health consequences, including damage to the brain, kidneys, and other organs. Cadmium is a toxic heavy metal that can accumulate in the body over time and the exposure to this element has been linked to a variety of adverse health effects. Cadmium toxicity can lead to an imbalance in the cellular redox state and be a source of oxidative stress. On the molecular level, cadmium ions negatively affect cellular metabolism, including the disruption of energy production, protein synthesis, and DNA damage. The study has been carried out on a group of 140 school-age children (8 to 14 years old) inhabiting the industrialized areas of Upper Silesia. The study population was divided into two sub-groups based on the median concentration of cadmium in blood (0.27 µg/L): Low-CdB and High-CdB. Measured traits comprised blood cadmium levels (CdB) as well as a blood count and selected oxidative stress markers. This research study aimed to demonstrate a correlation between the impact of exposure to elevated cadmium concentrations in a population of children and certain markers of oxidative stress, and 25-OH vitamin D3 concentration. A negative correlation has been found between cadmium concentration and 25-OH vitamin D3 level, protein sulfhydryl groups content in blood serum, glutathione reductase activity, and lipofuscin and malondialdehyde levels in erythrocytes. The concentration of 25-OH vitamin D3 in the High-CdB group was decreased by 23%. The oxidative stress indices can be considered a valuable indicator of early Cd-toxicity effects to be included in the routinely-applied cadmium exposure monitoring parameters, allowing the evaluation of stress intensity to the cell metabolism.

Vitamin B-6 and riboflavin, their metabolic interaction, and relationship with MTHFR genotype in adults aged 18–102 years

The generation of the active form of vitamin B-6, pyridoxal 5'-phosphate (PLP), in tissues is dependent upon riboflavin as flavin mononucleotide, but whether this interaction is important for maintaining vitamin B-6 status is unclear. To investigate vitamin B-6 and riboflavin status, their metabolic interaction, and relationship with methylenetetrahydrofolate reductase (MTHFR) genotype in adulthood. Data from 5612 adults aged 18-102 y were drawn from the Irish National Adult Nutrition Survey (NANS; population-based sample) and the Trinity-Ulster Department of Agriculture (TUDA) and Genovit cohorts (volunteer samples). Plasma PLP and erythrocyte glutathione reductase activation coefficient (EGRac), as a functional indicator of riboflavin, were determined. Older (≥65 y) compared with younger (<65 y) adults had significantly lower PLP concentrations (P<0.001). A stepwise decrease in plasma PLP was observed across riboflavin categories, from optimal (EGRac ≤1.26), to suboptimal (EGRac: 1.27-1.39), to deficient (EGRac ≥1.40) status, an effect most pronounced in older adults (mean±SEM: 76.4±0.9 vs 65.0±1.1 vs 55.4±1.2nmol/L; P<0.001). In individuals with the variant MTHFR 677TT genotype combined with riboflavin deficiency, compared with non-TT (CC/CT) genotype participants with sufficient riboflavin, we observed PLP concentrations of 52.1±2.9 compared with 76.8±0.7nmol/L (P<0.001). In participants with available dietary data (i.e., NANS cohort, n = 936), PLP was associated with vitamin B-6 intake (nonstandardized regression coefficient β: 2.49; 95% CI 1.75, 3.24; P<0.001), supplement use (β: 81.72; 95% CI: 66.01, 97.43; P<0.001), fortified food (β: 12.49; 95% CI: 2.08, 22.91; P=0.019), and EGRac (β: -65.81; 95% CI: -99.08, -32.54; P<0.001), along with BMI (β: -1.81; 95% CI: -3.31, -0.30; P=0.019). These results are consistent with the known metabolic dependency of PLP on flavin mononucleotide (FMN) and suggest that riboflavin may be the limiting nutrient for maintaining vitamin B-6 status, particularly in individuals with the MTHFR 677TT genotype. Randomized trials are necessary to investigate the PLP response to riboflavin intervention within the dietary range. The TUDA study and the NANS are registered at www.ClinicalTrials.gov as NCT02664584 (27 January 2016) and NCT03374748 (15 December 2017), respectively.Clinical Trial Registry details: Trinity-Ulster-Department of Agriculture (TUDA) study, ClinicalTrials.gov no. NCT02664584 (January 27th 2016); National Adult Nutrition Survey (NANS), ClinicalTrials.gov no. NCT03374748 (December 15th 2017).

Impact of the MTHFR C677T polymorphism on blood pressure and related central haemodynamic parameters in healthy adults.

BackgroundThe C677T polymorphism in the gene‐encoding methylenetetrahydrofolate reductase (MTHFR) is associated with an increased risk of hypertension and cardiovascular disease. Riboflavin, the MTHFR cofactor, is an important modulator of blood pressure (BP) in adults homozygous for this polymorphism (TT genotype). The effect of this genetic variant on BP and related central haemodynamic parameters in healthy adults has not been previously investigated and was examined in this study.MethodsBrachial BP, central BP and pulse wave velocity (PWV, SphygmoCor XCEL) were measured in adults aged 18–65 years prescreened for MTHFR genotype. Riboflavin status was assessed using the erythrocyte glutathione reductase activation coefficient assay.ResultsTwo hundred and forty‐two adults with the MTHFR 677TT genotype and age‐matched non‐TT (CC/CT) genotype controls were identified from a total cohort of 2546 adults prescreened for MTHFR genotype. The TT genotype was found to be an independent determinant of hypertension (p = 0.010), along with low‐riboflavin status (p = 0.002). Brachial systolic and diastolic BP were higher in TT versus non‐TT adults by 5.5 ± 1.2 and 2.4 ± 0.9 mmHg, respectively (both p < 0.001). A stronger phenotype was observed in women, with an almost 10 mmHg difference in mean systolic BP in TT versus non‐TT genotype groups: 134.9 (95% confidence interval [CI] 132.1–137.6) versus 125.2 (95% CI 122.3–128.0) mmHg; p < 0.001. In addition, PWV was faster in women with the TT genotype (p = 0.043).ConclusionThis study provides the first evidence that brachial and central BP are significantly higher in adults with the variant MTHFR 677TT genotype and that the BP phenotype is more pronounced in women.

No Association Between Riboflavin and Choline Status in a Sample of Canadian and Malaysian Women of Reproductive Age

ObjectivesRiboflavin and choline are essential nutrients that are metabolically inter-related and play major roles in one-carbon metabolism. Betaine can donate a methyl group to homocysteine, forming dimethylglycine (DMG) and methionine in the liver and kidney. Betaine is synthesized from choline in two irreversible reactions, the first of which is catalyzed by flavin adenine dinucleotide (FAD), a cofactor formed from riboflavin. In animal models, riboflavin status has been shown to influence choline and its metabolites betaine and DMG but there are no human studies. Here we examine whether riboflavin status modifies plasma choline, betaine, and DMG concentrations in healthy women (19–45 y).MethodsFasting blood was collected from 206 Canadian and 210 Malaysian women between 2015 and 2016. Riboflavin status was assessed using a functional biomarker, erythrocyte glutathione reductase activity coefficient (EGRac). Plasma choline, betaine, and DMG were determined by LCLC MS/MS. Plasma folate and vitamin B12 were determined using the microbiological method and an immune assay, respectively. General linear models were used to assess the independent relationship between EGRac and each of the choline metabolites with adjustment for potential confounders.Results71% of Malaysian women had EGRac ≥1.40 compared to 40% of Canadian women. Betaine, DMG, and vitamin B12 concentrations were significantly higher among Malaysian women compared to Canadian women (40.6 vs. 37.0 mmol/L, 2.7 vs. 2.4 mmol/L and 360 vs. 307 pmol/L, respectively). There were no significant associations between EGRac or riboflavin deficiency (defined as EGRac ≥1.40) and choline or its’ metabolites after adjustments for age, ethnicity, body mass index, plasma folate, and vitamin B12. In the adjusted models, plasma choline was positively associated with vitamin B12 concentrations (B = 0.002, 95% CI: 0.000, 0.003) and plasma betaine was positively associated with plasma folate (B = 0.18, 95%CI: 0.07, 0.29) and vitamin B12 (B = 0.011, 95%CI: 0.002, 0.020).ConclusionsOverall, riboflavin status was not associated with choline and its metabolites in Canadian and Malaysian women.Funding SourcesDairy Farmers of Canada, Saudi Arabian Cultural Bureau in Canada, and BC Children's Hospital Research Institute.

Daily supplementation of a multiple micronutrient powder improves folate but not thiamine, riboflavin, or vitamin B12 status among young Laotian children: a randomized controlled trial

PurposeTo assess the effects of intervention with a daily multiple micronutrient powder (MNP) on thiamine, riboflavin, folate, and B12 status among young Laotian children.MethodsChildren (n = 1704) aged 6–23 mo, participating in a double-blind placebo-controlled randomized trial were individually randomized to receive daily either MNP (containing 0.5 mg of thiamine, 0.5 mg riboflavin, 150 μg folic acid, and 0.9 μg vitamin B12 along with 11 other micronutrients) or placebo and followed for ~ 36 weeks. In a randomly selected sub-sample of 260 children, erythrocyte thiamine diphosphate (eThDP), plasma folate and B12 concentrations, and erythrocyte glutathione reductase activation coefficient (EGRac; riboflavin biomarker) were assessed at baseline and endline.ResultsThere was no treatment effect on endline eThDP concentrations (110.6 ± 8.9 nmol/L in MNP vs. 109.4 ± 8.9 nmol/L in placebo group; p = 0.924), EGRac (1.46 ± 0.3 vs. 1.49 ± 0.3; p = 0.184) and B12 concentrations (523.3 ± 24.6 pmol/L vs. 515.9 ± 24.8 pmol/L; p = 0.678). Likewise, the prevalence of thiamine, riboflavin, and B12 deficiencies did not differ significantly between the two groups. However, endline folate concentration was significantly higher in the MNP compared to the placebo group (28.2 ± 0.8 nmol/L vs 19.9 ± 0.8 nmol/L, respectively; p < 0.001), and correspondingly, the prevalence of folate deficiency was significantly lower in the MNP group (1.6% vs 17.4%; p = 0.015).ConclusionsCompared to a placebo, daily MNP for 9 months increased only folate but not thiamine, riboflavin, or B12 status in young Laotian children.Trial registrationThe trial was registered at www.clinicaltrials.gov (NCT02428647) on April 29 2015.

Association of plasma folic acid levels with parameters of glutathione metabolism in hypertensive patients with comorbid diseases

Objective. Hypertension (HTN) is characterized by a high incidence of comorbidity. An individual approach to the management of such patientsrequires new markers for differential diagnosis. The pathophysiology of most comorbidities is closely related to endothelial dysfunction triggered by an imbalance in redox processes.The aim of this work is to assess the relationship between plasma level of folic acid (FA) and the state of the redox system of erythrocyte glutathione in patients with HTN and concomitant target organ damage in cardiovascular diseases (HTN + CVD), chronic kidney disease (HTN + CKD) and discirculatory encephalopathy (HTN + DE).Design and methods. We enrolled 93 patients with HTN admitted to the clinics of the Pavlov University, and 30 donors of the reference group. We assessed plasma concentration of folic acid, the content of reduced glutathione (GSH) and the activity of glutathione reductase (GR) in erythrocytes.Results. In HTN patients with the deficiency of FA, a lower content of GSH and activity of GR in erythrocytes were found compared to the HTN patients without deficiency and the reference group. In the groups HTN + CVD and HTN + DE, the GSH level and GR activity correlated with the plasma concentration of FA and were lower in the subgroups with FA deficiency. In subgroups with normal FA content, these parameters did not differ from the reference indicators.Conclusions. The parameters of GSH and GR in patients with HTN + CVD and HTN + DE, but not HTN + CKD, can be considered as potential markers of functional FA deficiency.

Activity of enzymes of glutamate, energy and glutathione metabolism in the first juvenile depression with attenuated symptoms of schizophrenia

To reveal clinical and biological correlations in patients with attenuated symptoms of schizophrenia in the first juvenile depression, namely, the correlation between SOPS and HDRS-21 scores and the levels of activities of glutathione, glutamate and energy metabolism enzymes in the blood of patients. The study included 81 young men, aged 16-25 years, with the first depressive episode (ICD-10 items F32.1, F32.2, F32.38, F32.8), from which the groups with predominantly attenuated positive symptoms (group 1, n=36) and predominantly attenuated negative symptoms (group 2, n=24), and a group without attenuated schizophrenia symptoms (group 3, n=21) were selected. The control group consisted of 20 mentally healthy men aged 19-25 years. Psychometric methods (SOPS and HDRS-21) and psychopathological methods were used. Activities of cytochrome c oxidase (COX), glutamate dehydrogenase (GDH), glutathione reductase (GR), and glutathione S-transferase (GST) enzymes were determined spectrophotometrically in blood cells. When compared with the control group, activities of platelet GDH, GR, and GST (before and after treatment) were significantly reduced in groups 1, 2, 3 (Mann-Whitney U-test, p<0.0002, p<0.001 and p<0.0001, respectively); the activity of erythrocyte GST was reduced in group 1, and the activities of erythrocyte GR and GST were reduced in group 3 (p<0.05). In group 1, baseline COX (before treatment) was positively correlated with post-treatment SOPS-N scores (R=0.580, p=0.0003), while baseline erythrocyte GR was negatively correlated with post-treatment HDRS-21 scores (R=-0.591, p=0.0004). In group 2, baseline GDH levels were positively correlated with post-treatment scores on SOPS-P (R=0.425, p=0.0384), SOPS-N (R=0.500, p=0.0129), SOPS total (R=0.526, p=0.0083) and HDRS-21 (R=0.479, p=0.0180). The discovery of clinical and biological correlations in groups of patients with attenuated symptoms of schizophrenia in the structure of juvenile depression contributes to understanding the pathogenetic mechanisms of the formation of a high clinical risk of psychosis and contributes to the search for markers of the initial stages of schizophrenia.

Association of genetic variants in the GPX1 and GPX4 genes with the activities of glutathione-dependent enzymes, their interaction with smoking and the risk of acute pancreatitis

Genetic factors and tobacco smoke exposure can be associated with an increased risk of acute pancreatitis (AP). The pathogenesis of AP is associated with intensive oxidative stress. Glutathione peroxidase (GPx) is one of many enzymes involved in the neutralization of free radicals. This study aimed to investigate the impact of SNP rs1050450 in the GPX1 gene and rs713041 in the GPX4 gene on the activity of total GPx in a group of AP patients and healthy subjects. It was found that AP can contribute to decreased GPx activity (in plasma and erythrocyte lysate) accompanied by an increased glutathione reductase (GR) activity and decreased glutathione (GSH) concentration in two groups, non-smokers and smokers. A decreased GPx activity in erythrocyte lysate of AP patients compared to healthy subjects was associated with the occurrence of the CC genotype for SNP rs1050450. It was noted an increased GPx activity and decreased GR activity in erythrocytes of non-smoking AP patients with the TT genotype compared to subjects with the CC and TC genotype for SNP rs713041. However, in the group of smoking AP patients with this genotype, GR activity was elevated compared to non-smokers, which was accompanied by increased GSH concentration. These results can indicate that smoking in the course of AP can change the involvement of antioxidants in dependence on the genotype for the examined SNPs. The CC genotype for SNP rs1050450 and the TT genotype for rs713041 increases the risk of AP recurrence, which may be associated with increased MDA concentration.

The use of antioxidant medicine as an adjunctive therapy in late-onset schizophrenia spectrum disorders

BACKGROUND. Imbalance in the activity of neurotransmitter systems, mitochondrial impairment, and oxidative stress are potential neurobiological factors in the schizophrenia development.&#x0D; AIM. To test the hypothesis about the possibility of a subgroup selection from patients with late-onset schizophrenia spectrum disorders, for which the use of ethylmethylhydroxypyridine succinate as an adjunctive to antipsychotic therapy would be the most effective in relation to symptoms that are relatively more pronounced in patients of this subgroup.&#x0D; MATERIAL AND METHODS. 43 patients (women and men) aged 4578 years with late-onset (after 40 years) schizophrenia spectrum disorders were examined using clinical psychopathological, psychometric, biochemical and statistical research methods. Enzymatic activities of cytochrome C-oxidase, glutamate dehydrogenase, glutathione reductase and glutathione-S-transferase were assessed in blood cells twice (on a patient admission to the hospital and after a 28-day treatment course).&#x0D; RESULTS. Criteria for a patient assignment to the subgroup for ethylmethylhydroxypyridine succinate prescription were: more prominent side effects of pharmacotherapy, predominance of anxiety-hypochondriac symptoms and a lesser severity of psychosis. Before the treatment starting, the subgroup to which ethylmethylhydroxypyridine succinate was prescribed significantly differed in the activity of platelet glutamate dehydrogenase (p=0.031), glutathione-S-transferase (p=0.005), and erythrocyte glutathione reductase (p=0.045). As a result of the treatment course, the severity of symptoms by which the patients receiving ethylmethylhydroxypyridine succinate significantly differed before the starting the treatment, became undistinguished from those in the rest examined patients. After the treatment course, no significant differences in enzymatic activities were found in patients treated with ethylmethylhydroxypyridine succinate from those in other patients.&#x0D; CONCLUSION. This study confirmed the clinical validity of using a medicine with antioxidant properties as an adjunctive therapy to the main treatment in the selected subgroup of patients.

Effectiveness of a fortified drink in improving B vitamin biomarkers in older adults: a controlled intervention trial

BackgroundOlder adults are reported to have sub-optimal B vitamin status; targeted food-based solutions may help to address this. The objectives of the OptiAge food intervention study were to develop and investigate the effectiveness of a B vitamin-fortified drink in improving B vitamin biomarkers in older Irish adults with a primary outcome of change in the B vitamin biomarker status.MethodsA double-blinded randomised controlled trial was performed in parallel at University College Dublin and Ulster University. Participants aged > 50 years were recruited following screening for exclusion criteria (i.e. taking medications known to interfere with B vitamin metabolism, supplements containing B vitamins, consuming > 4 portions of B vitamin-fortified foods per week or diagnosed with gastrointestinal, liver or pulmonary disease). Recruited participants meeting the inclusion criteria were randomised (by sex and study centre) to receive daily for 16 weeks either B vitamin-fortified or placebo drinks as developed by Smartfish, Norway. Each B vitamin-fortified drink (200 ml) contained 200 µg folic acid, 10 µg vitamin B12, 10 mg vitamin B6 and 5 mg riboflavin, while the placebo was an identical, isocaloric formulation without added B vitamins. Fasting blood samples were collected pre- and post-intervention which were used to measure the primary outcome of change in B vitamin biomarker levels.ResultsA total of 95 participants were randomised, of which 81 commenced the trial. Of these, 70 completed (37 in the active and 33 in the placebo groups). Intention to treat (ITT) analysis of the B vitamins demonstrated a significant improvement in all B vitamin biomarkers in the active compared to placebo groups: p < 0.01 for each of serum folate, serum vitamin B12 and plasma pyridoxal 5′-phosphate (vitamin B6) and the functional riboflavin biomarker, erythrocyte glutathione reductase activation coefficient (EGRac). Correspondingly, a significant lowering of serum homocysteine from 11.9 (10.3–15.1) µmol/L to 10.6 (9.4–13.0) µmol/L was observed in response to the active treatment (P < 0.001). Similar results were seen in a per-protocol analysis.ConclusionsThe results demonstrate that a B vitamin-fortified drink was effective in optimising B vitamin status, making this a useful intervention option to improve B vitamin status in older adults.Trial registration ISRCTN, ISRCTN61709781—Retrospectively registered, https://www.isrctn.com/ISRCTN61709781

INFLUENCE OF MELATONIN ON AGE-RELATED CHANGES IN CARBOHYDRATE METABOLISM AND ANTIOXIDANT CAPACITY IN THE BLOOD OF ALLOXAN DIABETIC RATS

Melatonin and its metabolites have potent antioxidant/anti-inflammatory properties, and they have proven to be highly effective in a variety of disorders linked to inflammation and oxidative stress. The object of this experimental research was to ascertain the influence of aging on the level of basal glycemia and activities of glucose-6-phosphate dehydrogenase [EC1.1.1.49], pyruvate kinase [EC 2.7.1.40] and glutathione reductase [EC1.6.4.2] in erythrocytes of alloxan diabetic rats on the background of melatonin injections. Methods: We used 100 male Wistar rats, two age groups: the - 2-month (adult), and II - 4-month (old). Alloxan diabetes was evoked via injecting the rats with a 5% solution of alloxan monohydrate intraperitoneally in a dose of 170 mg/kg. Four days after diabetes induction, rats were divided into diabetic (untreated) and melatonin-diabetic group (10 mg/kg, daily and intraperitoneally for six weeks). Blood was taken from the tail vein evaluate the basal glycemia on 5-th and 47-th day after the injection of alloxan. Rats were sacrificed at the 47-th day of the experiment accordance with the ethical treatment of animals. Determinations of the enzymes activities were by standard methods. Statistical analysis was performed using Statistica 10 StatSoft Inc. Results. The level of basal glycemia on the fifth day of the experiment in animals of both groups increased on average by 115% from baseline values. We founded that on 47-th day this index was higher in group of old rats on 20% more than in adult rats. Pyruvate kinase activity in erythrocytes of adult and old animals with diabetes decreased by 34% and 51% respectively compared with the control. glucose-6-phosphate dehydrogenase activity in erythrocytes of adult and old animals with diabetes decreased by 25% and 44% respectively compared with the control on 47-th day. The changes may be the result of age-related disorders of glucose metabolism due to disturbances in free radical mechanisms. Glutathione reductase activity in erythrocytes of adult and old animals with diabetes decreased by 30% and 36% respectively compared with the control on 47-th day. A 42-days injection of melatonin to the alloxan diabetic rats of both groups contributed to a normalization of the level of basal glycemia, the activities of pyruvate kinase and glutathione reductase in the rat blood, as well as to a considerable increase of the activity of glucose-6-phosphate dehydrogenase, whose level exceeded by average 9% this particular index in the control group of animals. Under the influence of melatonin increase activity of glucose-6-phosphate dehydrogenase in the blood of rats may be due to the increasing number of substrate for glucose-6-phosphate dehydrogenase (stimulating the flow of glucose into cells and its phosphorylation) and direct action. Conclusion. In this case melatonin probably increases use of glucose for regeneration of NADPH2 and aerobic oxidation of glucose that indicate an acceleration of antioxidative protection and energy production in blood of adult and old diabetic rats.

State of antioxidant system in urogenital trichomoniasis and membranotropic effect of metronidazole

The objective of this work was to study the activity of glutathione peroxidase, glutathione reductase and the level of sulfhydryl groups in erythrocytes of patients with urogenital trichomoniasis and the effect of metronidazole on the degree of osmotic and peroxide resistance of erythrocytes from healthy donors. We examined 15 patients with urogenital trichomoniasis and 20 healthy volunteers. We studied native preparations, and also carried out a culture method using the Johnson-Trussel nutrient medium (CPLM) to identify Trichomona vaginalis. The activity of glutathione reductase, glutathione peroxidase and the level of total sulfhydryl groups were determined in erythrocytes of peripheral blood. The membrane effect of metronidazole was evaluated in in vitro experiment by the degree of osmotic and peroxide resistance of erythrocytes from healthy people. It has been established that a significant decrease in glutathione reductase and glutathione peroxidase activities in erythrocytes is observed, which indicates a violation of the antioxidant system in this pathology. It was shown in vitro experiment, that metronidazole in low concentration (80 μmol /l) has the ability to inhibit erythrocyte hypotonic hemolysis, and high concentration (250 μmol/l) leads to a decrease in osmotic and peroxide resistance of erythrocytes. Thus, inhibition of the activity of the enzymatic link of the antioxidant defense is observed in urogenital trichomoniasis, which is one of the mechanisms for the development of pathology at the cellular level in this disease. It has been shown that the isolated membranotropic action of metronidazole depends on its concentration – the drug at low concentration is able to inhibit hypotonic hemolysis of erythrocytes, and high concentration makes them more sensitive to the osmotic and peroxide hemolysis. The obtained results should be taken into account in the development of complex methods of therapy for urogenital trichomoniasis.

Dietary Riboflavin Intake and Riboflavin Status in Young Adult Women Living in Metro Vancouver, Canada.

ABSTRACTBackgroundNutrition surveys suggest that <10% of Canadian adults have inadequate riboflavin intakes. However, biochemical riboflavin deficiency [erythrocyte glutathione reductase activity coefficient (EGRac) ≥1.40] has been reported in 41% of young adult women living in Metro Vancouver. Canadian Chinese ethnicity comprise >25% of Vancouver's population and are postulated to have poorer riboflavin status than those of European ethnicity because they could be less likely to consume dairy products and fortified wheat.ObjectivesThe objectives of this study were to determine dietary riboflavin intake and food sources, and to assess the association between riboflavin intake and status in young women of European (n = 107) and Chinese (n = 91) ethnicities living in Metro Vancouver, Canada.MethodsThis was a cross-sectional study conducted in women (aged 19–45 y). Women were healthy, not pregnant or breastfeeding, of European or Chinese ethnicities, and not taking riboflavin-containing supplements for the past 4 mo. Dietary riboflavin intake was assessed using the past-year Diet History Questionnaire II, and riboflavin status (EGRac) was measured in fasting venous blood samples.ResultsOnly 7% of participants had dietary riboflavin intakes below the Estimated Average Requirement (0.9 mg/d), but 40% of women had biochemical riboflavin deficiency (EGRac ≥1.40). Although more Canadian women of European ethnicity than Chinese ethnicity had biochemical riboflavin deficiency (46% and 34%; P < 0.001), median dietary riboflavin intake did not differ (1.73 and 1.82 mg/d; P = 0.587). Dairy products and vegetables contributed the most to riboflavin intake. Energy-adjusted dietary riboflavin intake was inversely associated with EGRac (B = −0.04, 95% CI: −0.07, −0.01). However, after further adjustment the relation was not significant.ConclusionsOverall, women of reproductive age living in Metro Vancouver, Canada, had a low prevalence of inadequate dietary riboflavin intake despite the high prevalence of apparent biochemical riboflavin deficiency.

The relationship of folate deficiency, hyperhomocysteinemia and glutathione metabolism in hypertensive patients

Hypertension (HTN) is often accompanied by folic acid (FA) deficiency and hyperhomocysteinemia (HHcy). Reduced glutathione (GSH) and dependent enzymes determine the state of cellular antioxidant and redox systems in cardiovascular pathology. The aim of our work is to assess the relationship between the status of FA and the presence of HHcy with enzymes of glutathione metabolism and the redox state of erythrocyte glutathione in HTN. Design and methods. In blood plasma samples from 43 HTN patients admitted to the clinic of Pavlov University, the concentration of FA and total homocysteine (oHcy) was determined. We also evaluated the level of GSH, the activity of glutathione peroxidase and glutathione reductase (GR) in erythrocytes. Results. In the whole group, GR activity positively correlated with the concentration of FA (R = 0,415; p = 0,001). A significant decrease in GR activity (U/g Hb) was found in the subgroup with the low level of FA [0,8 (0,5–1,1)] compared with the subgroup without a FA deficiency [1,2 (0,9–2,0)]. The GSH level (μM/g Hb) was also lower (p &lt; 0,018) in the subgroup with FA deficiency [1,3 (0,9–2,1)] compared with the subgroup with normal FA levels [1,8 (1,5–4,6)]. A significant decrease in the level of GSH and GR activity in the subgroup with HHcy was found compared with the corresponding parameters in the subgroup without HHcy. However, even in the absence of HHcy patients with FA deficiency demonstrated a significant decrease in GR activity compared to patients without FA deficiency. In this case, GR positively correlated with FA (R = 0,564; p = 0,03). Conclusions. The deficiency of FA can increase the deficiency of GR activity, regardless of the level of oHcy. The indicator of GR activity in erythrocytes can be considered as a possible marker of functional deficiency of FA in the absence of HHcy.

A Short-Term Resistance Training Circuit Improved Antioxidants in Sedentary Adults with Down Syndrome.

Previous studies have found aerobic training improved oxidative damage in people with Down syndrome (DS). However, there is a lack of information regarding the influence of resistance training on redox imbalance in this population. Accordingly, this study was conducted to determine the effect of resistance training (RT) on antioxidant defence system in sedentary adults with DS. Thirty-six male adults with DS were recruited through different community support groups. Eighteen were randomly assigned to perform a circuit RT program with 6 stations, 3 days/week for 12 weeks. Plasma total antioxidant status (TAS), reduced glutathione (GHS), ascorbate, serum α-tocopherol, and erythrocyte glutathione reductase activity were assessed. Plasma malondialdehyde (MDA) and carbonyl groups (CG) were assessed as markers of oxidative damage. Muscle strength was also measured. Dynamic torque of knee extensors and flexors as well as maximal handgrip strength was significantly improved after the completion of the training program. Plasma levels of TAS and erythrocyte glutathione reductase (GR) activity were significantly increased. Conversely, MDA and CG levels were significantly reduced. It was concluded RT improved antioxidant defence system and reduced oxidative damage in adults with DS. Further, long-term studies are required to determine whether the increased antioxidant system may improve clinical outcomes of adults with DS.

A 2-Year Randomized Controlled Trial With Low-Dose B-Vitamin Supplementation Shows Benefits on Bone Mineral Density in Adults With Lower B12 Status.

Folate, vitamins B12, B6, and riboflavin are required for one-carbon metabolism and may affect bone health, but no previous randomized trial has investigated all four nutrients in this context. We investigated the effect of low-dose B-vitamins for 2 years on bone mineral density (BMD) in a dual-centered, 2-year randomized controlled trial (RCT) in adults aged ≥50 years. Eligible participants not consuming B-vitamin supplements or fortified foods >4times weekly were randomized to receive daily either combined folic acid (200 μg), vitamin B12 (10 μg), vitamin B6 (10 mg), and riboflavin (5mg), or "active" placebo, whereby both the intervention and placebo groups received vitamin D (10 μg). BMD was assessed before and after intervention using dual-energy X-ray absorptiometry (DXA) scanning of the total hip, femoral neck, and lumbar spine (L1 to L4). Of 205 eligible participants randomized, 167 completed the trial in full. B-vitamin intervention resulted in increases in serum folate (p < 0.001), serum B12 (p < 0.001), and plasma pyridoxal-5-phosphate (p < 0.001) and decreases in functional biomarkers of B-vitamin status, erythrocyte glutathione reductase activation coefficient (p < 0.001), serum methylmalonic acid (MMA; p < 0.001), and serum total homocysteine (p < 0.001). B-vitamin intervention had no overall effect on BMD, which declined in both treatment groups by approximately 1% (ranging from -0.7% to -1.4%). However, in participants with lower baseline B12 status (serum B12 <246 pmol/L or MMA ≥0.22 μmol/L), B-vitamin intervention reduced the 2-year BMD decline versus placebo: adjusted mean (95% confidence interval [CI]) change of -0.003 (-0.008, 0.002) versus -0.015 (-0.021, -0.010)g/cm2 at the total hip and -0.004 (-0.010, 0.001) versus -0.013 (-0.018, -0.007)g/cm2 at the femoral neck. In conclusion, the findings indicate that although low-dose B-vitamin intervention for 2 years had no overall effect on BMD, improving B-vitamin status appears to have specific benefits for bone health in adults with lower B12 status. © 2022 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

AGE-RELATED CHANGES OF GLYCOLYTIC ACTIVITY AND ANTIOXIDANT CAPACITY IN THE BLOOD OF ALLOXAN DIABETIC RATS

The increasing incidence of type 1 diabetes coupled with advances in treatment of type 1 diabetes has resulted in an unprecedented number of older adults living with and controllable type 1 diabetes. The objective of this experimental study was to assess the impact of aging on the level of basal glycaemia and activities of glucose-6-phosphate dehydrogenase [EC1.1.1.49], pyruvate kinase [EC 2.7.1.40] and glutathione reductase [EC1.6.4.2] in erythrocytes of alloxan-diabetic rats. Methods: We used 100 male Wistar rats, divided into two age groups: I group included- 2-month (adult) animals, and II group was made up of 4-month (old) animals. Diabetes was modelled by injecting the rats with 5% solution of alloxan monohydrate intraperitoneally in a dose of 170 mg/kg. Blood was taken from the tail vein to evaluate the basal glycaemia on 5-th and 47-th day after the alloxan injection. Rats were sacrificed on the 47-th day of the experiment in accordance with the regulations on ethical treatment of vertebrates. The assessment of the activity of the enzymes was carried out by standard methods. Statistical analysis was performed by using Statistica 10 StatSoft Inc. Results. The level of basal glycaemia on the fifth day of the experiment in the animals of both groups went up on average by 115% from baseline values. We founded that on 47-th day this index was higher in group of old rats by 20% than in adult rats. Pyruvate kinase activity in erythrocytes of adult and old animals with diabetes decreased by 35% and 50% respectively compared with the control. Glucose-6-phosphate dehydrogenase activity in erythrocytes of adult and old animals with diabetes decreased by 27% and 45% respectively compared with the control on 47-th day. The changes may be considered as the result of age-related disorders of glucose metabolism due to disturbances in free radical mechanisms. Glutathione reductase activity in erythrocytes of adult and old animals with diabetes decreased by 29% and 35% respectively compared with the control on 47-th day. Conclusion. We have determined when getting aged, the alloxan-diabetic rats demonstrate changes in the sensitivity of pyruvate kinase, glucose-6-phosphate dehydrogenase and glutathione reductase activities in erythrocytes resulted from the effect of diabetes mellitus factors (hyperglycaemia). We can suggest that glycaemic control is key purpose for older patients with type 1 diabetes in order to prevent of complication, which can be aggravated with age.