Plasma Glutathione Levels Research Articles

Making bloodwork work: the impact of sample collection, processing, and storage on plasma glutathione measurement, and implications for translation

Psychiatry has traditionally focused on the study of neurons and neurotransmitter physiology in the pathophysiology and treatment of psychiatric disorders. A growing literature highlights REDOX imbalance (a state in which demand for antioxidants surpasses their bioavailability) as a common pathophysiology for a diverse array of brain conditions (e.g., trichotillomania, schizophrenia, autism, Parkinson’s disease). REDOX imbalance is typically measured via plasma glutathione, as glutathione is critical to the adaptive antioxidant response in the brain. Accordingly, glutathione, its precursors, and/or metabolites serve as biomarkers of disease risk, therapeutic targets, and measures of treatment response. However, as with any emerging field, there are currently several different methods for collection, processing, storage, and calculation of summary measures of plasma glutathione metabolism, within and between preclinical and clinical research. The lack of evidence-based best-practice methodology hampers reproducibility (preclinical or clinical), and translation (between preclinical and clinical work). To address this methodological need, here we used a repeated measures within-subject design to investigate how sample preparation (type of anticoagulant used during blood collection, deproteinization status, and storage temperature) affects plasma glutathione levels. Accordingly, we collected whole blood from mice (N = 13), and then, using a commercially available kit, quantified glutathione in plasma stored in four different ways. Presuming that these preparation conditions and post-processing calculations are unimportant, we would expect to see no difference in glutathione levels and summary measures from the same sample. However, we found each of these variables to significantly alter quantified glutathione levels. Accordingly, we propose a vital, gold-standard methodology for both sample collection, processing, and storage of plasma used for glutathione quantification and for summary calculations of glutathione that can be used preclinically and clinically, thus yielding more streamlined translation.

Potential Protective Effect of Hesperidin (Vitamin P) against Glyphosate-Induced Spermatogenesis Damage in Male Rats: Biochemical and Histopathological Findings on Reproductive Parameters.

The aim of this study was to investigate the effectiveness of hesperidin (HES) on testicular histopathological changes, biochemical changes, and semen characteristics in rats exposed to glyphosate (GLP). The control group was given a normal diet devoid of GLP and HES, the HES group was given 100 mg/kg/day HES with the normal diet, the GLP group was given GLP at the LD50/10 dose of normal feed, which was 787.85 mg/kg/day, and the GLP + HES group was given normal feed containing 787.85 mg/kg/day LD50/10 dose of GLP in addition to 100 mg/kg/day HES. GLP administration reduced sperm motility, sperm plasma membrane integrity, glutathione levels, and total antioxidant levels in the testicular tissues of rats. Moreover, it caused an increase in right testis and left epididymis weights, abnormal sperm counts, malondialdehyde levels, total oxidant status, and DNA damage. The HES treatment showed curative effects on these parameters. Furthermore, HES was effective in lessening the histopathological damage that was caused by GLP. The results showedthat HES protects spermatological parameters and DNA integrity, improves antioxidant defenses, and lowers the damage and lipid peroxidation caused by GLP in testicular tissue.

Abstract Tu020: Contrasting effect of Ox-PAPC and PC-KLH in atherosclerotic plaques or peripheral blood T cell, macropahges and dendritic cell activation

Background: Atherosclerosis is a chronic inflammation characterized by accumulation of immune cells, oxidized low density lipoprotein (OxLDL) and dead cells in the lesions, both of the latter have the hapten phosphorylcholine (PC) as an important antigen. PC is also exposed on microorganisms and other compounds and needs a carrier to be antigenic. Antibodies against PC (anti-PC) ameliorates deleterious effects of both OxLDL and dead cells. Oxidized-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphatidylcholine (Ox-PAPC) is a major compound in the lipid moiety of OxLDL exposing PC. Methods: We studied effect of Ox-PAPC and PC with keyhole limpet cyanid (KLH) on T cells, dendritic cells (DC) and macrophages (MQ). We examined the influence of Ox-PAPC and PC conjugated to KLH (PC-KLH) on T cells, DC, and MQ from peripheral blood or atherosclerotic plaques. We investigated phospholipid and glutathione (GSH) levels in the blood plasma of healthy controls (n=30) and patients (n=30). Results: Ox-PAPC, in contrast to PAPC and PC-KLH, independently of DC, induced proinflammatory T cell activation, which was inhibited by mito tempo, inhibitor of reactive oxygen species (ROS). Ox-PAPC triggered inflammatory activities while reducing metabolic activities in T cells. PC-KLH, on the other hand, reduced CD86 and CD11C, proinflammatory cytokines, and induced anti-inflammatory IL-10 in DC. Ox-PAPC induced ROS, GSH, and proinflammatory MQ activation, inhibited by Mito tempo or GSH inhibitor. Additionally, Ox-PAPC, in the presence of PCSK9, elevated CD86 expression and lipid peroxidation. Patients exhibited higher GSH levels comapre to control groups, associated with phospholipid levels. Conclusions: The study reveals the differential effects of Ox-PAPC on various immune cell types, showcasing T cell activation independent of DC and heightened macrophage activation in the presence of PCSK9. Ox-PAPC-induced redox imbalance regulates T cell activation, apoptosis, and MQ activation, potentially rendering plaques more vulnerable and contributing to lesion formation and thus plaque rupture. In contrast, PC-KLH had anti-inflammatory effects, inducing anti-inflammatory cytokines from T cells, DC and MQ. PC-effects thus vary from proinflammatory to anti-inflammatory depending on presentation. Inhibition of OxPAPC effects by agents such as anti-PC could thus be anti-atherogenic, while on the other hand PC presented on a protein as KLH could be anti-atherogenic and anti-inflammatory itself.

Levels of melatonin and some indicators of angiogenesis, antioxidant system and lipid peroxidation in blood plasma in women with uterine leiomyoma

Uterine leiomyoma is one of the most common gynaecological diseases, which often leads to loss of fertility. It is known that in this pathology, damage to the tissues of the uterus is accompanied by oxidative stress, and the mechanisms of its compensation play a decisive role in the process of myometrial regeneration, especially when performing organ-preserving operations. The aim of the study was to determine the levels of melatonin, reproductive hormones and state of angiogenesis, antioxidant system and lipid peroxidation in women of reproductive age who were diagnosed with uterine leiomyoma. Material and methods. 60 women of reproductive age with uterine leiomyoma (study group) were examined. The control group consisted of 20 healthy women of the same age group. Melatonin levels in women's blood were determined once, on an empty stomach, at 8 a.m.; the concentrations of estradiol and progesterone in the follicular phase of the menstrual cycle were also measured. The state of the angiogenesis system was studied by examining the levels of vascular endothelial growth factor (VEGF) and the content of the final metabolites of nitric oxide NO in blood plasma. The activity of the antioxidant system was assessed by blood plasma concentrations of reduced glutathione, glutathione peroxidase, and glutathione transferase. Indicators of lipid peroxidation were investigated by the content of malondialdehyde (MDA) in blood plasma and erythrocytes. Results. It was found that in the patients of the study group with leiomyoma, there was a significantly lower level of melatonin in blood plasma (111.1 ± 18.5 ng/mL) compared to the control group (153.5 ± 8.5 ng/mL), while the concentration of estradiol was almost three times higher (107.4 ± 25.3 pg/mL) compared to the control group (36.2 ± 3.2 pg/mL), and the concentration of progesterone was 1.9 times higher (2.1 ± 0.4 ng/mL compared to 1.1 ± 0.5 ng/mL in the control group). The level of VEGF in blood plasma in women with uterine leiomyoma was also higher (90.4 ± 23.6 pg/mL) compared to the control group (35.1 ± 8.3 pg/mL), as well as the concentration of final metabolites of nitric oxide, which reached 25.3 ± 5.9 pg/mL compared to 9.9 ± 3.9 pg/mL in the control group. The reduced glutathione level in the blood plasma of women with uterine leiomyoma was significantly lower (0.77 ± 0.13 µmol/L) compared to healthy women (1.02 ± 0.14 µmol/L) in the control group, while the concentrations of glutathione-S-transferase and glutathione peroxidase enzymes were higher (161.3 ± 22.3 ng/mL and 235.7 ± 35.9 ng/mL, respectively), whereas in the control group these indicators were 118.9 ± 18.0 ng/mL and 105.3 ± 41.2 ng/mL, respectively. The MDA content in women of the study group was higher, measuring 5.2 ± 0.8 nmol/L in plasma and 10.8 ± 1.1 nmol/L in erythrocytes compared to 2.3 ± 0.8 nmol/L and 5.3 ± 0.8 nmol/L in the control group, respectively. Conclusions. The levels of melatonin and reduced glutathione in the blood plasma of women with uterine leiomyoma were significantly lower, while the concentrations of estradiol, progesterone, glutathione-S-transferase, glutathione peroxidase, VEGF, final metabolites of nitric oxide, as well as the average MDA content in plasma and erythrocytes, were significantly higher compared to healthy women. In the study group, the relationship between the level of progesterone in blood plasma and the content of malondialdehyde in erythrocytes was described using a linear regression equation, which, as we suggest, indicates an activating effect of progesterone on oxidative stress mechanisms in the myometrium. In our opinion, oxidative stress in uterine leiomyoma occurs due to inadequate activity of the antioxidant system, an integral part of which is melatonin, leading to enhanced angiogenesis against the background of estrogen-induced myometrial proliferation.

Plasma Redox Balance in Advanced-Maternal-Age Pregnant Women and Effects of Plasma on Umbilical Cord Mesenchymal Stem Cells.

Pregnancy at advanced maternal age (AMA) is a condition of potential risk for the development of maternal-fetal complications with possible repercussions even in the long term. Here, we analyzed the changes in plasma redox balance and the effects of plasma on human umbilical cord mesenchymal cells (hUMSCs) in AMA pregnant women (patients) at various timings of pregnancy. One hundred patients and twenty pregnant women younger than 40 years (controls) were recruited and evaluated at various timings during pregnancy until after delivery. Plasma samples were used to measure the thiobarbituric acid reactive substances (TBARS), glutathione and nitric oxide (NO). In addition, plasma was used to stimulate the hUMSCs, which were tested for cell viability, reactive oxygen species (ROS) and NO release. The obtained results showed that, throughout pregnancy until after delivery in patients, the levels of plasma glutathione and NO were lower than those of controls, while those of TBARS were higher. Moreover, plasma of patients reduced cell viability and NO release, and increased ROS release in hUMSCs. Our results highlighted alterations in the redox balance and the presence of potentially harmful circulating factors in plasma of patients. They could have clinical relevance for the prevention of complications related to AMA pregnancy.

Dietary cystine restriction increases the proliferative capacity of the small intestine of mice.

Currently, over 88 million people are estimated to have adopted a vegan or vegetarian diet. Cysteine is a semi-essential amino acid, which availability is largely dependent on dietary intake of meat, eggs and whole grains. Vegan/vegetarian diets are therefore inherently low in cysteine. Sufficient uptake of cysteine is crucial, as it serves as substrate for protein synthesis and can be converted to taurine and glutathione. We found earlier that intermolecular cystine bridges are essential for the barrier function of the intestinal mucus layer. Therefore, we now investigate the effect of low dietary cystine on the intestine. Mice (8/group) received a high fat diet with a normal or low cystine concentration for 2 weeks. We observed no changes in plasma methionine, cysteine, taurine or glutathione levels or bile acid conjugation after 2 weeks of low cystine feeding. In the colon, dietary cystine restriction results in an increase in goblet cell numbers, and a borderline significant increase mucus layer thickness. Gut microbiome composition and expression of stem cell markers did not change on the low cystine diet. Remarkably, stem cell markers, as well as the proliferation marker Ki67, were increased upon cystine restriction in the small intestine. In line with this, gene set enrichment analysis indicated enrichment of Wnt signaling in the small intestine of mice on the low cystine diet, indicative of increased epithelial proliferation. In conclusion, 2 weeks of cystine restriction did not result in apparent systemic effects, but the low cystine diet increased the proliferative capacity specifically of the small intestine and induced the number of goblet cells in the colon.

A Single Dose of Microencapsulated Cocoa Supplementation Does Not Speed up Muscle Force Recovery after Eccentric Exercise-Induced Muscle Damage: A Placebo-Controlled, Double-Blind, Crossover Study

Exercise-induced muscle damage is associated with symptoms such as inflammation, delayed-onset muscle soreness, and impaired muscle performance. The intake of cocoa polyphenols has been suggested to improve muscle recovery due to their antioxidant and anti-inflammatory capacity. However, their bioavailability presents a challenge. Therefore, food microencapsulation may be an alternative to protect polyphenols, ensuring their biological effects. This study aimed to investigate the effect of a single dose of microencapsulated cocoa on the changes in muscle damage markers after eccentric exercise. In this randomized, double-blind, crossover study, fourteen healthy volunteers with previous resistance training experience performed 6 × 10 maximal isokinetic eccentric contractions of their elbow flexors using an isokinetic dynamometer after ingesting 25 g of microencapsulated cocoa or placebo. Peak isometric torque was measured using maximal voluntary isometric contractions, and pain was measured using a visual analogic scale both before and 24 h, 48 h, and 72 h after the damage protocol. Plasma glutathione and malondialdehyde levels were measured using high-performance liquid chromatography, and concentrations of myoglobin and C-reactive protein were determined using a fluorescence immunoassay analyzer. Significant decreases were seen in the peak isometric torque and pain measures from pre- to 72 h post-eccentric exercise. A significant main effect for time was found only for plasma myoglobin at 2 h, 48 h, and 72 h, and for C-reactive protein at 2 h, compared to the pre-eccentric exercise values. No significant time-treatment effects were observed (all p > 0.05). This study demonstrated that microencapsulated cocoa cannot improve muscle recovery after eccentric exercise, at least when a single dose is consumed.

Cerebroprotein hydrolysate attenuates neurodegenerative changes in Alzheimer's mice model via ferroptosis pathway.

Introduction: Cerebroprotein hydrolysate has been proven to improve cognitive function in patients with Alzheimer's disease (AD). We explored the safety and effectiveness of the clinical administration of oral cerebroprotein hydrolysate in AD, and possible mechanisms related to the neuronal ferroptosis pathway. Methods: Three-month-old male APP/PS1 double-transgenic mice were randomly divided into AD model (n = 8) and intervention (n = 8) groups. Eight non-transgenic wild-type (WT) C57 mice were used as age-matched controls. The experiments were started at the age of 6 months. The intervention group was then administered cerebroprotein hydrolysate nutrient solution (11.9mg/kg/day) via chronic gavage, the other groups received an identical volume of distilled water. Behavioural experiments were performed after 90days of continuous administration. Serum and hippocampal tissues were then collected for histomorphological observation, tau and p-tau expression, and ferroptosis markers analysis. Results: Cerebroprotein hydrolysate simplified movement trajectories and shortened escape latencies of APP/PS1 mice in the Morris water maze test. Neuronal morphologies were restored in hippocampal tissues on haematoxylin-eosin staining. In the AD-model group, Aβ protein and p-tau/tau expression levels were elevated, plasma Fe2+ and malondialdehyde levels were elevated, GXP4 protein expression and plasma glutathione levels declined than controls. All indices improved after cerebroprotein hydrolysate intervention. Conclusion: Cerebroprotein hydrolysate improves learning and memory function, alleviates neuronal damage, and reduces the deposition of pathological AD markers in AD mice, which may be related to the inhibition of neuronal ferroptosis.

Dietary Supplements Potentially Target Plasma Glutathione Levels to Improve Cardiometabolic Health in Patients with Diabetes Mellitus: A Systematic Review of Randomized Clinical Trials.

Cardiovascular diseases (CVDs) continue to be the leading cause of death in people with diabetes mellitus. Severely suppressed intracellular antioxidant defenses, including low plasma glutathione (GSH) levels, are consistently linked with the pathological features of diabetes such as oxidative stress and inflammation. In fact, it has already been established that low plasma GSH levels are associated with increased risk of CVD in people with diabetes. Dietary supplements are widely used and may offer therapeutic benefits for people with diabetes at an increased risk of developing CVDs. However, such information remains to be thoroughly scrutinized. Hence, the current systematic review explored prominent search engines, including PubMed and Google Scholar, for updated literature from randomized clinical trials reporting on the effects of dietary supplements on plasma GSH levels in people with diabetes. Available evidence indicates that dietary supplements, such as coenzyme Q10, selenium, curcumin, omega-3 fatty acids, and vitamin E or D, may potentially improve cardiometabolic health in patients with diabetes. Such beneficial effects are related to enhancing plasma GSH levels and reducing cholesterol, including biomarkers of oxidative stress and inflammation. However, available evidence is very limited and additional clinical studies are still required to validate these findings, including resolving issues related to the bioavailability of these bioactive compounds.

Black Mulberry (Morus Nigra L.) juice has positive effects on spermatological parameters in male New Zealand White rabbits

The current study aimed to investigate the possible effects of black mulberry juice (BMJ) consumption on spermatologic parameters, oxidant/antioxidant activity, and seminal plasma testosterone concentrations of male rabbits. Twenty-four healthy adult New Zealand White rabbits were randomly divided into 4 groups (n = 6). By oral gavage, 3.0 mL of distilled water, 0.75 mL BMJ + 2.25 distilled water, 1.50 mL BMJ + 1.5 mL distilled water, and 3 mL BMJ per 1 kg body weight were given daily for 9 weeks to rabbits in the C, LD, MD, and HD groups, respectively. Ejaculates were taken at the beginning and end of the experiments. Progressive motility, sperm concentration, and intact sperm rates were significantly increased especially in MD and HD groups. A significant decrease in malondialdehyde concentrations was apparent due to BMJ consumption. Moreover, there was a marked increase in catalase and glutathione peroxidase levels in the seminal plasma of the MD and HD groups. The current results indicated that, although seminal plasma protein levels and seminal plasma glutathione levels were not altered, overall oral BMJ treatments were able to improve sperm quality and antioxidant states of the male rabbits.

Inhibition of MRP1 Induces Fetal Hemoglobin through NRF2 Activation to Protect Human Erythroid Cells from Sickling

Introduction: Sickle cell disease (SCD) is a genetic disorder caused by a single nucleotide substitution (Glu6→Val) in the HBB gene coding for the β-globin of the hemoglobin A (HbA; α2β2) in erythroid cells. This point mutation leads to abnormal hemoglobin (HbS), which polymerizes in low oxygen condition and, ultimately, causes sickling of circulating erythrocytes leading to hemolysis, vascular inflammation, and painful vaso-occlusive crises. Hereditary persistence of fetal hemoglobin (HbF; α2γ2) in sickle cell patients significantly reduces clinical symptoms by preventing polymerization of HbS and subsequent cell sickling. Therefore, raising HbF expression in SCD patients is a promising therapeutic strategy. However, hydroxyurea, the standard of care for SCD and only FDA-approved HbF inducer, carries concerns for carcinogenicity-associated effects and the non-responders rate for this drug is relatively high (~30%). Consequently, there is a large unmet need for new therapies in SCD. Methods: To discover potential new HbF inducers, we conducted a genome-wide association study of HbF levels in 11,004 healthy donors from the INTERVAL study. We identified ABCC1, which encodes the multidrug resistant protein 1 (MRP1) and efflux pump for glutathione, as a putative causal gene modulating HbF expression. We detected a specific missense variant in ABCC1 which increases HbF expression in erythrocytes and strongly associates with plasma glutathione levels (β=0.8SD, P=3x10-38). To elucidate the role of MRP1 in HbF regulation, we performed a CRISPR-Cas9-mediated ABCC1 knockdown in erythroid HUDEP-2 cells and CD34+ hematopoietic stem cells and quantified the expression of HbF by flow cytometry, Western blot and high-performance liquid chromatography (HPLC) after erythroid differentiation (2-phase culture). We also generated HUDEP-2 clones homozygous for the p.R433S variant using CRISPR-Cas9 gene editing and measured HbF levels using the methods previously described. In CD34+ cells, MRP1 effect on HbF expression was also evaluated by using the selective MRP1-inhibitor, MK571. Transcriptomic, proteomics and phospho-proteomic analysis were performed to identify the mechanism of action involved in HbF induction after MRP1 inhibition. Expression of erythroid differentiation markers GATA1, ALAS2, LRF and Band 3 was assessed by Western blot to evaluate the impact of MRP1 inhibition on erythroid differentiation. Finally, we conducted sickling assays on MK571-treated CD34+ cells from SCD patients. Results:ABCC1 knockdown (90% gene silencing compared to control) in HUDEP-2 cells increased intracellular glutathione level (7−fold, P<0.01) and HbF (2.5−fold, P<0.05), as shown by flow cytometry, HPLC and Western blot analysis, after 7-day differentiation. Similarly, HUDEP-2 clones homozygous for the ABCC1-R433S allele presented an increase in intracellular glutathione (6-fold, P<0.05) and HbF expression levels (6-fold, P<0.01) compared to non-treated cells. In CD34+ cells knockdown for ABCC1 (95% gene silencing compared to control), HbF was increased (1.5−fold, P<0.05) after 14-day differentiation. Furthermore, pharmacological inhibition of MRP1, by MK571, in CD34+ cells led to an induction in glutathione (5-fold, P<0.01) and HbF (2-fold, P<0.05), and prevented sickling of CD34+ cells from SCD patients (2-fold decrease, P<0.05) in hypoxic condition (2% O2), without impacting erythroid differentiation. Finally, transcriptomic, proteomic, and phospho-proteomic integrated analyses in MK571-treated CD34+ cells revealed an activation of the NRF2-mediated oxidative stress pathway. Addition of ML385 (10µM), an NRF2 inhibitor, in MK571-treated CD34+ cells completely abolished HbF induction, confirming a NRF2-dependent HbF induction. Conclusion: In summary, we identified MRP1 as a HbF modulator in human erythroid cells and demonstrated that MRP1 inhibition induces HbF and prevents sickling of CD34+ cells through NRF2-mediated oxidative stress pathway activation without affecting erythroid differentiation, establishing MRP1 as a potential new therapeutic target for SCD. Future directions include replication of the ABCC1 genetic association in an independent cohort, and in vivo studies aiming to confirm the efficacy of MRP1 inhibition on improving SCD disease biomarkers.

Ongoing oxidative stress in individuals with post-acute sequelae of COVID-19.

Coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 infection is associated with lower plasma glutathione (GSH) levels due to oxidative stress. However, plasma levels may not reflect brain GSH levels. Individuals with post-acute sequelae of COVID-19 (PASC) have a higher prevalence of cognitive fatigue, which might be related to altered brain γ-aminobutyric-acid (GABA) levels. Hence, our study aims to measure the brain GSH and GABA levels in PASC. 29 PASC participants and 24 uninfected controls were recruited for this study. Each was evaluated with detailed neuropsychiatric assessments and an edited proton MRS (Hadamard Encoding and Reconstruction of Mega-Edited Spectroscopy, HERMES) method to measure GABA and GSH concentrations in predominantly grey matter (GM) and predominantly white matter (WM) brain frontal voxels. PASC participants were 219 ± 137 days since their COVID-19 diagnosis. Nine individuals with PASC were hospitalized. Compared to controls, individuals with PASC had similar levels of GABA in both brain regions, but lower GSH and greater age-related GSH decline in the frontal GM region. The lower-than-normal frontal GM GSH level in participants with PASC suggest that they have ongoing oxidative stress in the brain, and that older individuals may be even more vulnerable to oxidative stress.

N-Acetylcysteine effects on glutathione and glutamate in schizophrenia: A preliminary MRS study

N-acetylcysteine (NAC) is a commonly used antioxidant that may have beneficial effects for schizophrenia. In this double-blind, randomized, placebo-controlled preliminary study, 40 patients with schizophrenia or schizoaffective disorder were randomized to receive 2400 mg NAC daily or placebo over eight weeks to examine the effects of NAC on prefrontal magnetic resonance spectroscopy levels of glutathione and glutamate. Secondary outcomes included negative symptoms, cognition, and plasma glutathione levels. We found that NAC treatment was associated with increased glutathione (statistically significant) and decreased glutamate (trend-level) compared with placebo in medial prefrontal cortex but not dorsolateral prefrontal cortex. We also observed a baseline association between medial prefrontal cortex levels of glutathione and plasma reduced / oxidized glutathione ratios. No treatment effects on symptoms or cognition were observed. Taken together, these findings indicate that treatment with N-acetylcysteine may increase medial prefrontal cortical levels of glutathione after eight weeks of treatment. These changes in cortical levels of glutathione may serve as an early biomarker of later clinical change and may underlie the cognitive and symptomatic improvements reported in longer-term treatment studies.

Dietary glycine supplementation alleviates transportation-induced stress in common carp, Cyprinus carpio

Fish transportation is a stressful event in aquaculture industry, which causes crowding stress, oxidative stress, hypoxia, and water pollution stress. Therefore, it is necessary to work on the methods for mitigation of such stresses. Glycine has been known for its roles in the antioxidant system and ammonia detoxification in fish. Therefore, the present study aimed at assessing the effects of dietary glycine on growth performance, stress and antioxidant responses of common carp, Cyprinus carpio, during transportation. The fish were fed diets supplemented with 0 (G0), 2.5 (G2.5), 5 (G5), and 10 (G10) g/kg glycine for 8 weeks, then transported in plastic bags for 6 h. According to the results, dietary glycine induced no significant changes in growth performance of the fish. Plasma ammonia decreased, but plasma urea increased by dietary glycine supplementation before and/or after transportation. Dietary glycine significantly increased plasma glutathione (GSH) levels before transportation and suppressed GSH decrease after transportation, particularly at 5 and 10 g/kg. Glycine supplementation significantly suppressed plasma cortisol, glucose, superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase, and malondialdehyde after transportation, particularly at 5 and 10 g/kg. The present study demonstrates that glycine is a useful dietary supplement for common carp, as it improves antioxidant capacity and helps the fish during transportation. Such effects can be achieved by dietary glycine supplementation at 5 g/kg in common carp.

Cysteine is a limiting factor for glioma proliferation and survival.

Nutritional intervention is becoming more prevalent as adjuvant therapy for many cancers in view of the tumor dependence on external sources for some nutrients. However, little is known about the mechanisms that make cancer cells require certain nutrients from the microenvironment. Herein, we report the dependence of glioma cells on exogenous cysteine/cystine, despite this amino acid being nonessential. Using several 13C‐tracers and analysis of cystathionine synthase and cystathioninase levels, we revealed that glioma cells were not able to support glutathione synthesis through the transsulfuration pathway, which allows methionine to be converted to cysteine in cysteine/cystine‐deprived conditions. Therefore, we explored the nutritional deprivation in a mouse model of glioma. Animals subjected to a cysteine/cystine‐free diet survived longer, although this increase did not attain statistical significance, with concomitant reductions in plasma glutathione and cysteine levels. At the end point, however, tumors displayed the ability to synthesize glutathione, even though higher levels of oxidative stress were detected. We observed a compensation from the nutritional intervention revealed as the recovery of cysteine‐related metabolite levels in plasma. Our study highlights a time window where cysteine deprivation can be exploited for additional therapeutic strategies.

The effect of vitamin D supplementation on tobacco-related disorders in individuals with a tobacco use disorder: a randomized clinical trial

Vitamin D deficiency in cigarette smokers (CS) might associate with several complications, including metabolic deficits, depression and anxiety. This study evaluated the effects of vitamin D on mental health symptoms, nicotine misuse, and biomarkers of metabolic diseases in individuals with a tobacco use disorder. A randomized, double-blind, placebo-controlled trial was conducted with 60 CS subjects receiving either 50,000 IU vitamin D supplements (n = 30) or placebo (n = 30) every 2 weeks for 24-weeks. Nicotine misuse, mental health scale, and metabolic parameters were measured before and after the intervention in the CS subjects. Compared with the placebo-group, after the 24-weeks intervention, serum 25 (OH) vitamin D levels increased in the intervention group (β 2.96; 95% CI, 0.91, 5.01; P = 0.006). In addition, vitamin D supplementation significantly improved Beck Depression Inventory (BDI) (β −2.06; 95% CI, −3.84, −0.28; P = 0.02). In addition, vitamin D administration significantly decreased fasting plasma glucose (FPG) (β −4.56; 95% CI, −8.94, −0.19; P = 0.04), insulin (β −0.50; 95% CI, −0.88, −0.13; P = 0.009), and homeostasis model of assessment-estimated insulin resistance (HOMA-IR) levels (β −0.21; 95% CI, −0.33, −0.08; P = 0.001). Furthermore, vitamin D resulted in a significant elevation in total antioxidant capacity (TAC) (β 81.20; 95% CI, 18.30, 144.11; P = 0.01), and plasma glutathione (GSH) levels (β 73.05; 95% CI, 18.56, 127.54; P = 0.01), compared with the placebo-group. Administration of vitamin D for 24-weeks to CS subjects had beneficial effects on symptoms of depression and several metabolic biomarkers. While this preliminary study suggests that vitamin D might have beneficial effects, its clinical efficacy in individuals with a tobacco use disorder should be further validated in future clinical trials.

Plasma Glutathione Levels Decreased with Cognitive Decline among People with Mild Cognitive Impairment (MCI): A Two-Year Prospective Study.

Glutathione (GSH) is a major endogenous antioxidant. Several studies have shown GSH redox imbalance and altered GSH levels in Alzheimer’s disease (AD) patients. Early detection is crucial for the outcome of AD. However, whether GSH can serve as a biomarker during the very early-phase of AD, such as mild cognitive impairment (MCI), remains unknown. The current prospective study aimed to examine the longitudinal change in plasma GSH concentration and its influence on cognitive decline in MCI. Overall, 49 patients with MCI and 16 healthy individuals were recruited. Plasma GSH levels and cognitive function, measured by the Mini-Mental Status Examination (MMSE) and Alzheimer’s disease assessment scale-cognitive subscale (ADAS-cog), were monitored every 6 months. We employed multiple regressions to examine the role of GSH level in cognitive decline in the 2 years period. The MCI patients showed significant decline in plasma GSH levels and cognitive function from baseline to endpoint (month 24). In comparison, the healthy individuals’ GSH concentration and cognitive function did not change significantly. Further, both GSH level at baseline and GSH level change from baseline to endpoint significantly influenced cognitive decline among the MCI patients. To our knowledge, this is the first study to demonstrate that both plasma GSH levels and cognitive function declined 2 years later among the MCI patients in a prospective manner. If replicated by future studies, blood GSH concentration may be regarded as a biomarker for monitoring cognitive change in MCI.

Effect of L-Carnitine and/or Calf Thymus Gland Extract Supplementation on Immunity, Antioxidant, Duodenal Histomorphometric, Growth, and Economic Performance of Japanese Quail (Coturnix coturnix japonica)

This study aimed to evaluate the supplementation (of drinking water) effects of L-Carnitine (LC) and/or calf thymus extract (TE) on immunity, antioxidant, duodenal histomorphometric, growth, and economic performance of Japanese quail. Six hundred, one day old unsexed quail were divided into four groups, the control group (G1) received plain drinking water, while G2, G3, and G4 were supplemented with LC and/or TE, respectively. The treated groups recorded a significant (p < 0.05) improvement in the final body weight, weight gain, feed conversion ratio, phagocytic activity %, phagocytic index, lymphocytes %, and plasma glutathione level compared to the G1. On the other hand, these supplementations significantly lower the mortality rate %, heterophil %, H/L ratio and plasma malondialdehyde level of the supplemented groups relative to G1. The supplemented groups recorded a non-significant increase in duodenal histomorphometric parameters (villous height, villous width, surface area, and crypt depth) relative to G1. Generally, LC and/or TE improve the values of economic profile (total returns, net profit, total return/total costs %, etc.) in treated groups compared to the control one. In conclusion, L-carnitine supplementation was more effective than TE or their combination with TE in the immunological, anti-oxidative, growth, and economic performance.

Relationship between coccidiosis and lipid peroxidation ‎level in blood of cattle.‎

‎ The recent paper studies the impact of coccidiosis on blood levels of cattle ‎since it is one of the parasites that attack both‏ ‏poultry and cattle epidemically and ‎infect the intestine leading to big economical loss as well as high deaths of those ‎animals. Due to the few number of relevant studies that present for the this parasite ‎effect on blood level as well as the levels of anti-oxidation, Glutathione, I did such ‎the study which includes 40 cows and 35 calves of 2-3 months of age from which a ‎stool sample was taken plus a 5 mml blood sample. Next, I diagnosed the infected ‎animal through testing the symptoms of those animals and examining the stool by ‎both floating and smear methods directly to find the ovarian cysts of coccidiosis. ‎Then, I determined the level of infection (the number of parasites per 1 gm of stool) ‎by using Macmaster way so the animals were divided into two groups (infected and ‎non-infected). After that, some biochemical tests were done to check the levels of ‎Malondialdehyde (MDA) as a final outcome of lipid peroxidation using colorful ‎TBA ase and to measure the absorption of ( 450 um) ‎‏ ‏wavelength. Also, I divided ‎the Glutathione (GSH) level in blood plasma using Elmanz method.‎

Anesthetic efficacy and hemato-biochemical effects of thymol on juvenile Nile tilapia, Oreochromis niloticus

In this study, two experiments were performed to investigate the anesthetic efficiency, hematological and biochemical effects of thymol on Nile tilapia, Oreochromis niloticus. In the first experiment, in order to estimate thymol anesthetic efficacy, tilapia juveniles were exposed to anesthetic baths at six different concentrations of thymol (10, 20, 40, 60, 80, and 100 mg L−1) and the induction and recovery times were recorded. The second experiment was performed with five treatments: CON (non-anesthetized fish), TH1 (anesthesia with 80 mg L−1 thymol within 1 min), TH5 (anesthesia with 30 mg L−1 thymol within 5 min), EU1 (anesthesia with 80 mg L−1 eugenol within 1 min), and EU5 (anesthesia with 30 mg L−1 eugenol within 5 min). The results showed that thymol at 10 mg L−1 concentration failed to induce anesthesia; however 20–100 mg L−1 concentrations induced anesthesia within 491–56.3 s. A strong negative relationship was detected between thymol concentrations and induction times (R2 = 0.906). Anesthesia had no significant effects on plasma albumin, triglyceride, cholesterol, alanine aminotransferase, and the blood mean corpuscular hemoglobin levels. The anesthetized fish exhibited significant elevations in plasma cortisol, glucose, lactate, and the blood hematocrit and mean corpuscular volume; these parameters exhibited significant elevations in the fish anesthetized by 30 mg L−1 anesthetics, compared to the 80 mg L−1. Plasma globulin, total protein, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, catalase, superoxide dismutase, and the blood hemoglobin and red blood cell significantly increased, as mean corpuscular hemoglobin concentration decreased, in the fish anesthetized by 30 mg L−1 anesthetics, compared to the CON fish. The anesthetized fish showed similar plasma glutathione levels, all significantly lower than the CON fish. The EU5 fish exhibited significant elevation in the plasma malondialdehyde levels, compared to the CON fish. In conclusion, thymol anesthetic efficacy was similar to eugenol in Nile tilapia weighing ~40 g at water temperature of 26 °C. Considering the physiological responses, slight but significant elevations in plasma proteins, lactate, lactate dehydrogenase, and malondialdehyde levels attest that thymol may induce less stress, hypoxia, and oxidative stress in the fish than eugenol, but further studies are needed to provide robust and clinically applicable data to support this hypothesis.