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Blood Advances Jun 2024The Glucose transporter 1 (GLUT1) is one of the most abundant proteins within the erythrocyte membrane and is required for glucose and dehydroascorbic acid (Vitamin C...
The Glucose transporter 1 (GLUT1) is one of the most abundant proteins within the erythrocyte membrane and is required for glucose and dehydroascorbic acid (Vitamin C precursor) transport. It is widely recognized as a key protein for red cell structure, function, and metabolism. Previous reports highlighted the importance of GLUT1 activity within these uniquely glycolysis-dependent cells, in particular for increasing antioxidant capacity needed to avoid irreversible damage from oxidative stress in humans. However, studies of glucose transporter roles in erythroid cells are complicated by species-specific differences between humans and mice. Here, using CRISPR-mediated gene editing of immortalized erythroblasts and adult CD34+ hematopoietic progenitor cells, we generate committed human erythroid cells completely deficient in expression of GLUT1. We show that absence of GLUT1 does not impede human erythroblast proliferation, differentiation, or enucleation. This work demonstrates for the first-time generation of enucleated human reticulocytes lacking GLUT1. The GLUT1-deficient reticulocytes possess no tangible alterations to membrane composition or deformability in reticulocytes. Metabolomic analyses of GLUT1-deficient reticulocytes reveal hallmarks of reduced glucose import, downregulated metabolic processes and upregulated AMPK-signalling, alongside alterations in antioxidant metabolism, resulting in increased osmotic fragility and metabolic shifts indicative of higher oxidant stress. Despite detectable metabolic changes in GLUT1 deficient reticulocytes, the absence of developmental phenotype, detectable proteomic compensation or impaired deformability comprehensively alters our understanding of the role of GLUT1 in red blood cell structure, function and metabolism. It also provides cell biological evidence supporting clinical consensus that reduced GLUT1 expression does not cause anaemia in GLUT1 deficiency syndrome.
PubMed: 38916993
DOI: 10.1182/bloodadvances.2024012743 -
Analytical Methods : Advancing Methods... Jun 2024Nanozymes have recently become a research hotspot because of the advantages of good stability, excellent catalytic performance and easy storage in comparison to natural...
Nanozymes have recently become a research hotspot because of the advantages of good stability, excellent catalytic performance and easy storage in comparison to natural enzymes. Nanozymes with oxidase-like activity get special attention because they needn't the participation of hydrogen peroxide. In this paper, poly(-isopropylacrylamide) nanogel with oxidase-like activity was synthesized for the first time. The catalytic mechanism was explored by EPR and UV spectroscopy after adding specific trapping agents of ROS, and the results showed that PNIPAM NG can catalyze O to O. In the presence of PNIPAM NG, -phenylenediamine (OPD) and ascorbic acid (AA) can be oxidized to 2,3-diaminophenazine (oxOPD) and dehydroascorbic acid (DHA), and DHA can further react with OPD to produce a fluorescence substance. The colorimetric and fluorescence detection platforms for AA were constructed based on the above principles. Both platforms have satisfactory results in real samples. The fluorescence platform has better sensitivity and selectivity than the colorimetric platform.
PubMed: 38910452
DOI: 10.1039/d4ay00638k -
Spectrochimica Acta. Part A, Molecular... Jun 2024Herein, a novel ratiometric strategy for ultra-sensitive detection of o-phenylenediamine (OPD) was proposed based on combinatorial reactions of in-situ fluorogenic...
Herein, a novel ratiometric strategy for ultra-sensitive detection of o-phenylenediamine (OPD) was proposed based on combinatorial reactions of in-situ fluorogenic reaction and in-situ formation of red fluorescent dithiothreitol-copper nanoparticles (DTT-CuNPs). Here, Cu is used both as an oxidant and as a precursor. Dehydroascorbic acid (DHAA) is formed via redox reaction of AA and Cu. Then, DHAA reacts with OPD to yield blue fluorescent quinoxaline (OXD) with emission peak at 434 nm through in-situ fluorogenic reaction. Red emitting DTT-CuNPs with emission peak at 666 nm is instantly generated due to the coordination reaction between DTT and the residual Cu which is not consumed by AA. The fluorescence intensity (FI) of OXD at 434 nm is closely relied on the concentration of OPD, which can be used as a response signal for OPD detection. Meanwhile, FI of DTT-CuNPs at 666 nm has no significant change, which can be used as a reference signal for OPD detection. Thus, the ratio (F/F) of the Cu/AA/DTT sensing system is successfully employed to quantify OPD, exhibiting a wide linear range from 0.2 µM to 60 µM, with LOD of 0.09 µM.
PubMed: 38906060
DOI: 10.1016/j.saa.2024.124671 -
Food Chemistry Jun 2024A validated silver nanoparticle assay (SNaP-C) for quantitation of Vitamin C, as ascorbic acid (AA) and total AA (TAA), was applied to 31 beverages. SNaP-C assay results...
A validated silver nanoparticle assay (SNaP-C) for quantitation of Vitamin C, as ascorbic acid (AA) and total AA (TAA), was applied to 31 beverages. SNaP-C assay results (LOD of 2.2 mg/L AA) were compared to AA and TAA determined by high-performance liquid chromatography with UV/Vis (LOD = 0.4 mg/L AA), and two well-known assays. All approaches were calibrated using meta-phosphoric acid stabilized AA, where the reducing agent tris(2-carboxyethyl) phosphine hydrochloride was added to convert dehydroascorbic acid to AA for determination of TAA. Statistical comparisons of these four resulting datasets were completed. SNaP-C and HPLC were not statistically significantly different (P > 0.05) for comparison of AA and TAA (mg/L) in these samples, whereas the CUPRAC and Folin-Ciocalteu assays statistically significantly overestimated values of AA and TAA content, respectively. The SNaP-C method is a novel assay that has high specificity for AA capable of quantifying TAA with addition of TCEP.
PubMed: 38905840
DOI: 10.1016/j.foodchem.2024.140112 -
Scientific Reports Jun 2024Body weight is an important economic trait for sheep meat production, and its genetic improvement is considered one of the main goals in the sheep breeding program....
Body weight is an important economic trait for sheep meat production, and its genetic improvement is considered one of the main goals in the sheep breeding program. Identifying genomic regions that are associated with growth-related traits accelerates the process of animal breeding through marker-assisted selection, which leads to increased response to selection. In this study, we conducted a weighted single-step genome-wide association study (WssGWAS) to identify potential candidate genes for direct and maternal genetic effects associated with birth weight (BW) and weaning weight (WW) in Baluchi sheep. The data used in this research included 13,408 birth and 13,170 weaning records collected at Abbas-Abad Baluchi Sheep Breeding Station, Mashhad-Iran. Genotypic data of 94 lambs genotyped by Illumina 50K SNP BeadChip for 54,241 markers were used. The proportion of variance explained by genomic windows was calculated by summing the variance of SNPs within 1 megabase (Mb). The top 10 window genomic regions explaining the highest percentages of additive and maternal genetic variances were selected as candidate window genomic regions associated with body weights. Our findings showed that for BW, the top-ranked genomic regions (1 Mb windows) explained 4.30 and 4.92% of the direct additive and maternal genetic variances, respectively. The direct additive genetic variance explained by the genomic window regions varied from 0.31 on chromosome 1 to 0.59 on chromosome 8. The highest (0.84%) and lowest (0.32%) maternal genetic variances were explained by genomic windows on chromosome 10 and 17, respectively. For WW, the top 10 genomic regions explained 6.38 and 5.76% of the direct additive and maternal genetic variances, respectively. The highest and lowest contribution of direct additive genetic variances were 1.37% and 0.42%, respectively, both explained by genomic regions on chromosome 2. For maternal effects on WW, the highest (1.38%) and lowest (0.41%) genetic variances were explained by genomic windows on chromosome 2. Further investigation of these regions identified several possible candidate genes associated with body weight. Gene ontology analysis using the DAVID database identified several functional terms, such as translation repressor activity, nucleic acid binding, dehydroascorbic acid transporter activity, growth factor activity and SH2 domain binding.
Topics: Animals; Genome-Wide Association Study; Weaning; Female; Polymorphism, Single Nucleotide; Sheep; Birth Weight; Quantitative Trait Loci; Body Weight; Maternal Inheritance; Breeding; Genotype; Male; Phenotype
PubMed: 38849438
DOI: 10.1038/s41598-024-63974-0 -
Journal of Food Protection Jul 2024Chlorine is commonly used by the fresh produce industry to sanitize water and minimize pathogen cross-contamination during handling. The pH of chlorinated water is often...
Chlorine is commonly used by the fresh produce industry to sanitize water and minimize pathogen cross-contamination during handling. The pH of chlorinated water is often reduced to values of pH 6-7, most commonly with citric acid to stabilize the active antimicrobial, hypochlorous acid (a form of free chlorine). Previous studies have demonstrated that citric acid reacts with chlorine to form trichloromethane, a major chlorine by-product in water and a potential human carcinogen. However, it is unclear if other pH control agents could be used in the place of citric acid to minimize the formation of trichloromethane. The objective of the present study was to determine the reactivity of organic and inorganic pH control agents, with chlorine, to generate trichloromethane. Free chlorine (∼100 mg/L) was mixed with 10 mM of each of twelve organic acids and two inorganic pH control agents (i.e., sodium acid sulfate and phosphoric acid) to effect a pH level of 6.5. Free chlorine and trichloromethane levels were measured over 3 h at 3 and 22°C. Results demonstrated that ascorbic acid, dehydroascorbic acid, citric acid, and malic acid rapidly depleted free chlorine concentrations at both 22°C and 3°C, while tartaric acid and lactic acid decreased chlorine concentrations more slowly. Other pH control agents did not significantly reduce free chlorine either at 22 or 3°C. Citric acid led to the generation of significantly higher concentrations of trichloromethane than did other acids. Chloroacetone was also found in chlorinated water in the presence of citric acid and ascorbic acid. Taking buffering capacity and pKa values into account, phosphoric acid and some organic acids may be used to replace citric acid as pH control agents in chlorinated water for washing fresh produce, to stabilize free chlorine level and reduce the generation of trichloromethane.
Topics: Chlorine; Hydrogen-Ion Concentration; Humans; Halogenation; Disinfectants
PubMed: 38734411
DOI: 10.1016/j.jfp.2024.100296 -
BMC Plant Biology May 2024In plants, GABA plays a critical role in regulating salinity stress tolerance. However, the response of soybean seedlings (Glycine max L.) to exogenous...
Gamma-aminobutyric acid (GABA) improves salinity stress tolerance in soybean seedlings by modulating their mineral nutrition, osmolyte contents, and ascorbate-glutathione cycle.
BACKGROUND
In plants, GABA plays a critical role in regulating salinity stress tolerance. However, the response of soybean seedlings (Glycine max L.) to exogenous gamma-aminobutyric acid (GABA) under saline stress conditions has not been fully elucidated.
RESULTS
This study investigated the effects of exogenous GABA (2 mM) on plant biomass and the physiological mechanism through which soybean plants are affected by saline stress conditions (0, 40, and 80 mM of NaCl and NaSO at a 1:1 molar ratio). We noticed that increased salinity stress negatively impacted the growth and metabolism of soybean seedlings, compared to control. The root-stem-leaf biomass (27- and 33%, 20- and 58%, and 25- and 59% under 40- and 80 mM stress, respectively]) and the concentration of chlorophyll a and chlorophyll b significantly decreased. Moreover, the carotenoid content increased significantly (by 35%) following treatment with 40 mM stress. The results exhibited significant increase in the concentration of hydrogen peroxide (HO), malondialdehyde (MDA), dehydroascorbic acid (DHA) oxidized glutathione (GSSG), Na, and Cl under 40- and 80 mM stress levels, respectively. However, the concentration of mineral nutrients, soluble proteins, and soluble sugars reduced significantly under both salinity stress levels. In contrast, the proline and glycine betaine concentrations increased compared with those in the control group. Moreover, the enzymatic activities of ascorbate peroxidase, monodehydroascorbate reductase, glutathione reductase, and glutathione peroxidase decreased significantly, while those of superoxide dismutase, catalase, peroxidase, and dehydroascorbate reductase increased following saline stress, indicating the overall sensitivity of the ascorbate-glutathione cycle (AsA-GSH). However, exogenous GABA decreased Na, Cl, HO, and MDA concentration but enhanced photosynthetic pigments, mineral nutrients (K, K/Na ratio, Zn, Fe, Mg, and Ca); osmolytes (proline, glycine betaine, soluble sugar, and soluble protein); enzymatic antioxidant activities; and AsA-GSH pools, thus reducing salinity-associated stress damage and resulting in improved growth and biomass. The positive impact of exogenously applied GABA on soybean plants could be attributed to its ability to improve their physiological stress response mechanisms and reduce harmful substances.
CONCLUSION
Applying GABA to soybean plants could be an effective strategy for mitigating salinity stress. In the future, molecular studies may contribute to a better understanding of the mechanisms by which GABA regulates salt tolerance in soybeans.
Topics: gamma-Aminobutyric Acid; Seedlings; Glycine max; Ascorbic Acid; Glutathione; Minerals; Salt Tolerance; Salt Stress; Chlorophyll; Salinity
PubMed: 38706002
DOI: 10.1186/s12870-024-05023-6 -
Analytical Chemistry Apr 2024Toward the challenges of signaling transduction amplified in enantioselective recognition, we herein devised an innovative strategy for highly selective recognition of...
Toward the challenges of signaling transduction amplified in enantioselective recognition, we herein devised an innovative strategy for highly selective recognition of amino acids and their derivatives, leveraging photothermal effects. In this approach, bifunctional l-ascorbic acid is employed to reduce silver ions on Au nanostars. Simultaneously, its oxidate (l-dehydroascorbic acid) is bonded to the silver shell as a chiral selector to prepare chiral nanoparticles (C-AuNS@Ag NPs) with the ability to recognize stereoisomers and sensitively modulate the photothermal effect. l-Dehydroascorbic acid can selectively capture one of the enantiomers of the two forms through hydrogen bonding and drive aggregation of the nanoparticles, which sharply enhances the photothermal effect. Consequently, the two forms of the system exhibit a significant temperature difference, which enables the discrimination and quantification of enantiomers. Our strategy verifies that six chiral amino acids and their derivatives can be discriminated with enantioselective response values of up to 79. Additionally, the chiral recognition mechanism was revealed through density functional theory (DFT) calculations, providing a paradigm shift in the development of enantiomeric recognition strategies.
PubMed: 38597814
DOI: 10.1021/acs.analchem.3c05751 -
Roczniki Panstwowego Zakladu Higieny 2024The aim of the study was to determine Vitamin C content in some fruits and vegetables (FAV) including apple, banana, orange, pineapple, watermelon, carrot and cucumber,...
OBJECTIVE
The aim of the study was to determine Vitamin C content in some fruits and vegetables (FAV) including apple, banana, orange, pineapple, watermelon, carrot and cucumber, sold in the local markets in Awka, Anambra State, Nigeria as well as Vitamin C content in two-component and three-component homogenates FAV. The work was also designed to investigate the dietary exposure and health effects of excess vitamin C intake in adults and children.
MATERIAL AND METHODS
Vitamin C as total ascorbic acid (AA) after reduction of dehydroascorbic acid was analyzed using both titrimetric and spectrophotometric methods. The titrimetric method involved iodometric back-titration while the spectrophotometric method was done at an absorbance of 530 nm. The dietary exposure was evaluated as the total FAV intake multiplied by chemical concentration in the FAV whereas the health effect of excess vitamin C intake was conducted using the hazard quotient (HQ).
RESULTS
The results revealed that Vitamin C for single fruits ranged from 11.76 - 41.17 mg/L for spectroscopic method and 16.9 - 31.84 mg/L for titrimetric method. Fruit homogenates showed Vitamin C concentrations of 14.70 - 220.58 mg/L and 17.23 - 209.09 mg/L for two-components homogenates: 29.41-132.35 mg/L and 31.05-113.10 mg/L for tri-components homogenates for spectrophotometric and titrimetric methods respectively. The results of dietary exposure and the health effects of excess vitamin C intake showed that children are more susceptible to health issues than adults in illnesses such as nausea, gastrointestinal pains, increased kidney stones and hyperactivity.
CONCLUSION
There is therefore the need for a national recommended dietary allowance for total ascorbic acid (AA) in FAV homogenates from a stakeholder point of view in Nigeria.
Topics: Adult; Child; Humans; Ascorbic Acid; Fruit; Vegetables; Dietary Exposure; Vitamins; Diet
PubMed: 38578155
DOI: 10.32394/rpzh.2024.0294 -
Animal Reproduction Science May 2024The incidence of bovine endometritis, which has a negative impact on the reproduction of dairy cows, has been recently increasing. In this study, the differential...
The incidence of bovine endometritis, which has a negative impact on the reproduction of dairy cows, has been recently increasing. In this study, the differential markers and metabolites of healthy cows and cows with endometritis were analyzed by measuring blood biochemical indicators and immune factors using biochemical and enzyme-linked immunosorbent assay kits combined with nontargeted metabolomics. The LC-QTOF platform was used to evaluate the serum metabolomics of healthy cows and cows with endometritis after 21-27 days of calving. The results showed that glucose, free fatty acid, calcium, sodium, albumin, and alanine aminotransferase levels were significantly lower in the serum of cows with endometritis than in healthy cows (P < 0.05). However, the serum potassium, interleukin-1, interleukin-6, and tumor necrosis factor levels were significantly higher in cows with endometritis (P < 0.05). In addition, the serum metabolome data analysis of the two groups showed that the expression of 468 metabolites was significantly different (P < 0.05), of which 291 were upregulated and 177 were downregulated. These metabolites were involved in 78 metabolic pathways, including amino acid, nucleotide, carbohydrate, lipid, and vitamin metabolism pathways; signal transduction pathways, and other biological pathways. Taken together, negative energy balance and immune activation, which are related to local abnormalities in amino acid, lipid, and carbohydrate metabolism, were the important causes of endometritis in dairy cows. Metabolites such as glucose, carnosine, dehydroascorbic acid, L-malic acid, tetrahydrofolic acid, and UDP-glucose may be used as key indicators in the hematological diagnosis and treatment of endometritis in dairy cows.
Topics: Female; Cattle; Animals; Endometritis; Cattle Diseases; Metabolomics; Biomarkers
PubMed: 38564886
DOI: 10.1016/j.anireprosci.2024.107460