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1.
Microbiome ; 9(1): 180, 2021 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-34470652

RESUMO

BACKGROUND: Probiotics have been used to regulate the gut microbiota and physiology in various contexts, but their precise mechanisms of action remain unclear. RESULTS: By population genomic analysis of 418 Bifidobacterium longum strains, including 143 newly sequenced in this study, three geographically distinct gene pools/populations, BLAsia1, BLAsia2, and BLothers, were identified. Genes involved in cell wall biosynthesis, particularly peptidoglycan biosynthesis, varied considerably among the core genomes of the different populations, but accessory genes that contributed to the carbohydrate metabolism were significantly distinct. Although active transmission was observed inter-host, inter-country, inter-city, intra-community, and intra-family, a single B. longum clone seemed to reside within each individual. A significant negative association was observed between host age and relative abundance of B. longum, while there was a strong positive association between host age and strain genotype [e.g., single nucleotide polymorphisms in the arginine biosynthesis pathway]. Further animal experiments performed with the B. longum isolates via using a D-galactose-induced aging mouse model supported these associations, in which B. longum strains with different genotypes in arginine biosynthesis pathway showed divergent abilities on protecting against host aging possibly via their different abilities to modify the metabolism of gut microbes. CONCLUSIONS: This is the first known example of research on the evolutionary history and transmission of this probiotic species. Our results propose a new mechanistic insight for promoting host longevity via the informed use of specific probiotics or molecules. Video abstract.


Assuntos
Bifidobacterium longum , Microbioma Gastrointestinal , Probióticos , Sulfaleno , Envelhecimento , Animais , Galactose , Microbioma Gastrointestinal/genética , Humanos , Camundongos
2.
Appl Microbiol Biotechnol ; 105(19): 7161-7170, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34515844

RESUMO

D-Galactose is an abundant carbohydrate monomer in nature and widely exists in macroalgae, plants, and dairy wastes. D-Galactose is useful as a raw material for biomass fuel production or low-calorie sweetener production, attracting increased attention. This article summarizes the studies on biotechnological processes for galactose utilization. Two main research directions of microbial fermentation and enzyme-catalyzed conversion from galactose-rich biomass are extensively reviewed. The review provides the recent discoveries for biofuel production from macroalgae, including the innovative methods in the pretreatment process and technological development in the fermentation process. As modern people pay more attention to health, enzyme technologies for low-calorie sweetener production are more urgently needed. D-Tagatose is a promising low-calorie alternative to sugar. We discuss the recent studies on characterization and genetic modification of L-arabinose isomerase to improve the bioconversion of D-galactose to D-tagatose. In addition, the trends and critical challenges in both research directions are outlined at the end. KEY POINTS: • The value and significance of galactose utilization are highlighted. • Biofuel production from galactose-rich biomass is accomplished by fermentation. • L-arabinose isomerase is a tool for bioconversion of D-galactose to D-tagatose.


Assuntos
Biotecnologia , Galactose , Catálise , Fermentação , Humanos
3.
J Med Food ; 24(9): 997-1009, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34524027

RESUMO

Oxidative stress has been demonstrated to be associated with numerous aging-related diseases. Ethyl acetate fraction of Abelmoschus manihot (L.) Medic (EA) had been reported to possess strong radical-scavenging activity due to its rich content of flavonoids. This work aimed to determine the protective effects of EA against oxidative injuries in vivo and in vitro, as well as to explore the relevant mechanisms behind these effects. Pretreatment with EA significantly elevated cell viability of H2O2-induced HepG2 cells, reduced the reactive oxygen species level, decreased apoptotic cells, and inhibited activities of caspase 3/9. Meanwhile, EA pretreatment elevated the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), while reduced malondialdehyde (MDA) generation and lactate dehydrogenase (LDH) release dose-dependently. In addition, EA modulated key marker genes expression of antioxidation and apoptosis-related signaling pathways at the messenger RNA (mRNA) and protein levels. In the animal studies, EA also significantly enhanced the antioxidant activity and reduced MDA generation in serum, liver, and brain of the D-galactose (D-gal)-induced mice. Furthermore, the histological analysis indicated that EA effectively alleviated liver and brain injury of mice induced by D-gal, dose-dependently. EA as a potential antioxidant agent promoted health and reduced the risk of aging-associated diseases.


Assuntos
Abelmoschus , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Abelmoschus/química , Acetatos , Envelhecimento , Animais , Antioxidantes/farmacologia , Flores/química , Galactose , Células Hep G2 , Humanos , Peróxido de Hidrogênio , Malondialdeído , Camundongos , Superóxido Dismutase/metabolismo
4.
Anal Chem ; 93(34): 11843-11851, 2021 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-34410102

RESUMO

A thermally reversible nanogel is used in capillary electrophoresis to create discrete regions for a galactosyltransferase reaction and separation. The ß1-4 galactosyltransferase enzyme, donor, and co-factor were patterned in the capillary. The substrate was driven through these zones and converted to galactosylated products, which were separated and identified. Using this capillary electrophoresis method, the degree of glycosylation was discernible for a pentasaccharide and for biantennary N-glycans. With the ability to distinguish between reaction products for which either one or two galactose residues were transferred, the capillary nanogel electrophoresis system was used to determine the Michaelis-Menten value, KM. For the ß1-4 galactosyltransferase, the KM value obtained for a pentasaccharide substrate was 1.23 ± 0.08 mM. Once KM was established, the enzyme/substrate ratio was evaluated to add a single galactose residue or to fully galactosylate a biantennary N-glycan. Additionally, capillary nanogel electrophoresis was adapted to transfer galactose residues to protein. The applicability of the method for real-time online modification of whole protein was demonstrated with the Herceptin glycoprotein. Complete retardation by Erythrina cristagalli lectin after enzymatic modification confirmed the addition of galactose residues to the Herceptin. This demonstrated the potential of the method to be used for online modification of other glycoproteins.


Assuntos
Galactose , Polissacarídeos , Eletroforese Capilar , Galactosiltransferases , Glicoproteínas , Nanogéis
5.
Bioresour Technol ; 340: 125697, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34358984

RESUMO

The effect of donor substrate and products partitioning on the performance of butyl-ß-galactoside synthesis with Aspergillus oryzae ß-galactosidase was studied. Firstly, the partition coefficient of the donor substrate (lactose) and the reaction products (glucose, galactose and butyl-ß-galactoside) were determined in the aqueous and organic phases of the reaction medium. In the temperature range studied (30 to 50 °C), butyl ß-galactoside was roughly 130 and 30-fold more soluble in the organic phase than lactose and the monosaccharides, respectively. Afterward, the effect of the 1-butanol/ aqueous phase ratio (α) on the reaction was evaluated in the range from 0.25 to 4. Results show that higher values of α reduce the incidence of secondary hydrolysis by favoring the extraction of butyl-ß-galactoside into the organic phase where it is not hydrolyzed, leading to higher yields. Also, major interfacial properties for butyl-ß-galactoside were determined at 25 °C.


Assuntos
Aspergillus oryzae , Galactose , Galactosídeos , Hidrólise , Lactose , beta-Galactosidase
6.
Int J Mol Sci ; 22(15)2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34360627

RESUMO

Fucosylation is involved in a wide range of biological processes from cellular adhesion to immune regulation. Although the upregulation of fucosylated glycans was reported in diseased corneas, its implication in ocular surface disorders remains largely unknown. In this study, we analyzed the expression of a fucosylated glycan on the ocular surface in two mouse models of dry eye disease (DED), the NOD.B10.H2b mouse model and the environmental desiccating stress model. We furthermore investigated the effects of aberrant fucosylation inhibition on the ocular surface and DED. Results demonstrated that the level of type 2 H antigen, an α(1,2)-fucosylated glycan, was highly increased in the cornea and conjunctiva both in NOD.B10.H2b mice and in BALB/c mice subjected to desiccating stress. Inhibition of α(1,2)-fucosylation by 2-deoxy-D-galactose (2-D-gal) reduced corneal epithelial defects and increased tear production in both DED models. Moreover, 2-D-gal treatment suppressed the levels of inflammatory cytokines in the ocular surface and the percentages of IFN-γ+CD4+ cells in draining lymph nodes, whereas it did not affect the number of conjunctival goblet cells, the MUC5AC level or the meibomian gland area. Together, the findings indicate that aberrant fucosylation underlies the pathogenesis of DED and may be a novel target for DED therapy.


Assuntos
Túnica Conjuntiva/metabolismo , Córnea/metabolismo , Síndromes do Olho Seco/etiologia , Galactose/análogos & derivados , Antígenos H-2/metabolismo , Animais , Túnica Conjuntiva/efeitos dos fármacos , Córnea/efeitos dos fármacos , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Síndromes do Olho Seco/tratamento farmacológico , Síndromes do Olho Seco/metabolismo , Fucose/metabolismo , Galactose/farmacologia , Galactose/uso terapêutico , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Polissacarídeos/metabolismo
7.
Elife ; 102021 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-34369878

RESUMO

Bimodal gene expression by genetically identical cells is a pervasive feature of signaling networks and has been suggested to allow organisms to hedge their 'bets' in uncertain conditions. In the galactose-utilization (GAL) pathway of Saccharomyces cerevisiae, gene induction is unimodal or bimodal depending on natural genetic variation and pre-induction conditions. Here, we find that this variation in modality arises from regulation of two features of the pathway response: the fraction of cells that show induction and their level of expression. GAL3, the galactose sensor, controls the fraction of induced cells, and titrating its expression is sufficient to control modality; moreover, all the observed differences in modality between different pre-induction conditions and among natural isolates can be explained by changes in GAL3's regulation and activity. The ability to switch modality by tuning the activity of a single protein may allow rapid adaptation of bet hedging to maximize fitness in complex environments.


Assuntos
Regulação Fúngica da Expressão Gênica , Variação Genética , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Transdução de Sinais/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Galactose/metabolismo , Aptidão Genética
8.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(3): 276-280, 2021 May.
Artigo em Chinês | MEDLINE | ID: mdl-34374240

RESUMO

Objective: To investigate the protective effects and mechanisms of the polysaccharide from Balanophora involucrata HK.f (BIH) on liver injury induced by D-galactose in rats. Methods: Sixty male SD rats were randomly divided into 5 groups: the control group (n=12), the D-gal group (n=12), the BIH-L treatment group (D-gal+50 mg/kg BIH, n=12), the BIH-M treatment group (D-gal+100 mg/kg BIH, n=12), and the BIH-H treatment group (D-gal+200 mg/kg BIH, n=12). The rats were injected into the back of the neck with D-gal of 100 mg/kg/d subcutaneously except for the control group. The BIH treatment group were divided into BIH-L group (50 mg/(kg·d)), BIH-M group (100 mg/(kg·d)), and BIH-H group (200 mg/(kg·d)), respectively. The rats in the BIH group were intragastrically administrated with the relative BIH solution, while the rats in the control and D-gal group were treated with saline solution for 42 days. The serum contents of ALT, AST and DBIL c were tested by automatic biochemical analyzer, the content of MDA was determined by thiobarbital acid method and the SOD activity was detected by xanthine oxidase method. Expressions of Caspase-3, Bax, and Bcl-2 in liver were measured by Western blot, and morphological changes by HE staining and immunohistochemistry. Results: The serum contents of ALT, AST and DBIL in the D-gal group were significantly increased compared with those in Con group (P<0.01) and were decreased in the BIH group as compared with the D-gal group (P<0.01). Cell apoptosis, the Caspase-3 and Bax levels, and the MDA content in the D-gal group were increased compared with those in the control group (P<0.01). And BIH treatment could attenuate these effects induced by D-gal. Meanwhile, the Bcl-2 level and SOD activity in the BIH group were increased compared with that in the D-gal group (P<0.05, 0.01). Conclusion: BIH can protective liver injury through reducing cell apoptosis and inhibiting oxidative stress.


Assuntos
Galactose , Fígado , Animais , Galactose/metabolismo , Galactose/toxicidade , Fígado/metabolismo , Masculino , Estresse Oxidativo , Polissacarídeos , Ratos , Ratos Sprague-Dawley
9.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(4): 433-438, 2021 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-34374266

RESUMO

Objective: To study the effects and mechanisms of astaxanthin combined with aerobic exercise on renal senescence of rat induced by D-galactose. Methods: Sixty 3-month-old SPF SD rats were divided into control group (C group), acute senescence group (S group), astaxanthin+acute senescence group (AS group), aerobic exercise+acute senescence group (ES group), astaxanthin+aerobic exercise+acute senescence group (AES group), by two-factor two-level 2×2 factorial design with 12 rats in each group. Acute senescence model of rat was establshed by intraperitoneal injection with 100 mg/(kg·d) D-galactose, and the intervention was conducted with 20 mg/(kg·d) astaxanthin and/or aerobic exercise with 60% VO2max for 6 weeks. The histopathological/ultrastructural changes of the kidney were observed by light microscope/electron microscope; the levels of SOD, γ-GCS and MDA were detected by ELISA, and LDF in kidney was determined by fluorescence colorimetry; the protein expression of Nrf2 signaling pathway was detected by immunohistochemistry. Results: Compared with AS and ES group, in AES group, the improvement of renal tissue morphology/ultrastructure was more significant; LDF was decreased significantly (P<0.01); SOD activity was significantly increased (P<0.01); γ-GCS was significantly higher than that of AS group, but not significantly different from that of ES group (P>0.05); there was no significant difference in MDA between groups (P>0.05); the levels of Nrf2 and p-Nrf2 were increased significantly (P<0.05, P<0.01); HO-1 was significantly higher than that of ES group(P<0.05), but not significantly different compared with that of AS group(P>0.05). Conclusion: Astaxanthin combined with aerobic exercise can delay aging process of kidney, its mechanism may be that the combination regulate the protein expression in Nrf2 signaling pathway, Ⅱ detoxifying enzymes and antioxidant enzyme activity, and improve oxidative stress in kidney of rat induced by D-galactose.


Assuntos
Galactose , Fator 2 Relacionado a NF-E2 , Envelhecimento , Animais , Rim/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Xantofilas
10.
Molecules ; 26(16)2021 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-34443300

RESUMO

Frutalin is a plant lectin with beneficial immunobiological action, although the access to its active form is still restricted. Moreover, there is a knowledge gap on isoform activity and glycosylation impact on its bioactivity, and recombinant production protocols were seen as ineffective. Here, a simpler and faster production and purification protocol was developed, attaining a yield of purified frutalin 3.3-fold higher than that obtained previously. Hemagglutination assays confirmed that this frutalin isoform could not agglutinate rabbit erythrocytes, while maintaining the native tetrameric structure, as indicated by DLS analysis, and strong interaction with methyl-alpha-galactose, in fluorescence spectroscopy studies. The cytotoxicity of the recombinant frutalin isoform was shown in a broad panel of human cancer cells: colon (HCT116), melanoma (A375), triple-negative breast cancer (MDA-MB-231), and ovarian (IGROV-1). Treatment with 8.5-11.8 µM TrxFTL reduced proliferation of all cancer cells to half in 48 h. This anti-proliferative effect encompasses the p53 pathway since it was significantly reduced in p53-null colon cancer cells (HCT116 p53-/-; GI50 of 25.0 ± 3.0 µM), when compared to the isogenic p53-positive cells (HCT116 p53+/+; GI50 of 8.7 ± 1.8 µM; p < 0.002). This recombinantly produced frutalin isoform has relevant cytotoxic effect and its biological activity is not dependent on glycosylation. The developed E. coli production and purification protocol generates high yield of non-glycosylated frutalin isoform with potent cytotoxic activity, enabling the development of novel anticancer p53-targeting therapies.


Assuntos
Galectinas/farmacologia , Neoplasias/patologia , Sequência de Aminoácidos , Animais , Antineoplásicos/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Difusão Dinâmica da Luz , Escherichia coli/metabolismo , Galactose/metabolismo , Galectinas/química , Galectinas/isolamento & purificação , Glicosilação/efeitos dos fármacos , Hemaglutinação/efeitos dos fármacos , Modelos Moleculares , Peso Molecular , Coelhos , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacologia , Espectrometria de Fluorescência
11.
Chemistry ; 27(56): 14100-14107, 2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34398494

RESUMO

Two major hurdles in NP-based catalysis are the aggregation of the NPs and their recycling. Immobilization of NPs onto a 2D support is the most promising strategy to overcome these difficulties. Herein, amphiphilicity-driven self-assembly of galactose-hexaphenylbenzene-based amphiphiles into galactose-decorated 2D nanosheet is reported. The extremely dense decoration of reducing sugar on the surface of the sheets is used for the in situ synthesis and immobilization of ultrafine catalytically active AgNPs by using Tollens' reaction. The potential of the system as a catalyst for the reduction of various nitroaromatics is demonstrated. Enhanced catalytic activity is observed for the immobilized AgNPs when compared to the corresponding discrete AgNPs. Recovery of the catalytic system from the reaction mixture by ultrafiltration and its subsequent recycling for several cycles without dropping its activity is shown. This is the first report demonstrating the in situ synthesis and immobilization of ultrafine AgNPs onto a 2D nanosheet that exhibits excellent catalytic performance for the reduction of nitroaromatics.


Assuntos
Galactose , Nanopartículas Metálicas , Catálise , Prata
12.
Int J Mol Sci ; 22(12)2021 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-34205463

RESUMO

Violacein is a naturally occurring purple pigment, widely used in cosmetics and has potent antibacterial and antiviral properties. Violacein can be produced from tryptophan, consequently sufficient tryptophan biosynthesis is the key to violacein production. However, the complicated biosynthetic pathways and regulatory mechanisms often make the tryptophan overproduction challenging in Escherichia coli. In this study, we used the adaptive laboratory evolution (ALE) strategy to improve violacein production using galactose as a carbon source. During the ALE, a tryptophan-responsive biosensor was employed to provide selection pressure to enrich tryptophan-producing cells. From the biosensor-assisted ALE, we obtained an evolved population of cells capable of effectively catabolizing galactose to tryptophan and subsequently used the population to obtain the best violacein producer. In addition, whole-genome sequencing of the evolved strain identified point mutations beneficial to the overproduction. Overall, we demonstrated that the biosensor-assisted ALE strategy could be used to rapidly and selectively evolve the producers to yield high violacein production.


Assuntos
Evolução Biológica , Técnicas Biossensoriais , Galactose/metabolismo , Indóis/metabolismo , Engenharia Metabólica , Escherichia coli , Proteínas de Escherichia coli , Triptofano/metabolismo
13.
Molecules ; 26(11)2021 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-34198754

RESUMO

BACKGROUND: There is an increasing need for botanicals to be used as an alternative and complementary medicine in the management of male infertility. Male infertility has been a major health/social challenge to people all over the world. This study, therefore, investigated the ameliorative potential of hydroethanolic leaf extract of Parquetina nigrescens (HELEPN) against d-galactose-induced testicular injury. METHODS: Thirty male Wistar rats were randomly allotted into six groups (n = 5). Group I (Normal control), Group II (300 mg/kg b.w. d-galactose), Group III and IV (250 and 500 mg/kg b.w. HELEPN, respectively), Group V and VI (both received 300 mg/kg b.w. of d-galactose with 250 and 500 mg/kg b.w of HELEPN, respectively). d-galactose administration started two weeks prior to HELEPN treatment which lasted for six weeks. All assays were carried out using established protocols. RESULTS: Administration of HELEPN at 250mg/kg and 500mg/kg concomitantly with d-galactose improved paired and relative testicular weights, levels of gonadotropins (LH and FSH) and testosterone, and poor sperm quality. HELEPN treatment reduced the levels of oxidative stress biomarkers (MDA, 8-OHDG, and AGEs) and inflammatory response (TNF-alpha and NO) to normal, as well as restoring the reduced activities of antioxidant enzymes (glutathione peroxidase, superoxide dismutase, and catalase). In addition, HELEPN treatment mitigated testicular DNA fragmentation and down-regulated caspase 3-activities. HELEPN at 500 mg/kg was observed to have the greatest ameliorative effect. CONCLUSION: HELEPN protects against d-galactose-induced testicular injury through antioxidative, anti-inflammatory, and antiapoptotic mechanisms.


Assuntos
Apocynaceae/química , Galactose/efeitos adversos , Infertilidade Masculina/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Testículo/lesões , Animais , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Etanol/química , Gonadotropinas/metabolismo , Humanos , Infertilidade Masculina/induzido quimicamente , Infertilidade Masculina/metabolismo , Masculino , Tamanho do Órgão/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Distribuição Aleatória , Ratos , Ratos Wistar , Análise do Sêmen , Testículo/efeitos dos fármacos , Testículo/metabolismo , Testosterona/metabolismo
14.
Nutrients ; 13(6)2021 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-34205338

RESUMO

This study investigated the antioxidant effects of whey protein peptide on learning and memory in aging C57BL/6N mice. A total of 72 SPF male C57BL/6N mice were used. Twelve mice were randomly selected as the control group, and the other mice were intraperitoneally injected with D-galactose (100 mg/kg body weight for 6 weeks), during which, the mice in the control group were intraperitoneally injected with the same amount of normal saline. After 6 weeks, the blood was taken from the epicanthus and the serum MDA level was measured, according to which, the mice were randomly divided into the model control group, the whey protein group (1.5 g/kg body weight), and three Whey protein peptide (WHP) intervention groups (0.3 g/kg body weight, 1.5 g/kg body weight, 3.0 g/kg body weight). The water solution of the test sample was administered by oral gavage every day. The intervention period was 30 days, during which, the model control group, the whey protein group, and the whey protein peptide group continued receiving intraperitoneal injections of D-galactose, while the control group continued receiving intraperitoneal injections of normal saline. After the intervention, behavioral experiments were conducted in the following order: open field test, water maze test, and new object recognition test. After the behavioral experiment, the morphology of hippocampal formation was observed by HE staining and TUNEL labeling. Oxidative stress-related indexes in the serum, liver, and brain were detected. Expression levels of the cholinergic system-related enzymes and proinflammatory cytokines in brain tissue were detected. Western blot was used to detect the expression of synaptic plasticity-related proteins in the mouse brain. The results showed that WHP could significantly improve the accumulation of MDA and PC, increase the activities of SOD and GSH-Px, resist oxidative stress injury, and enhance the potential of endogenous antioxidant defense mechanisms. WHP can significantly improve the decline of aging-related spatial exploration, body movement, and spatial and non-spatial learning/memory ability. Its specific mechanism may be related to reducing the degeneration of hippocampal nerve cells, reducing the apoptosis of nerve cells, improving the activity of AChE, reducing the expression of inflammatory factors (TNF-α and IL-1ß) in brain tissue, reducing oxidative stress injury, and improving the expression of p-CaMKⅡ and BDNF synaptic plasticity protein. These results indicate that WHP can improve aging-related oxidative stress, as well as learning and memory impairment.


Assuntos
Envelhecimento/fisiologia , Antioxidantes/farmacologia , Aprendizagem/efeitos dos fármacos , Memória/efeitos dos fármacos , Peptídeos/administração & dosagem , Proteínas do Soro do Leite/química , Envelhecimento/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Encéfalo/fisiologia , Galactose/administração & dosagem , Hipocampo/citologia , Marcação In Situ das Extremidades Cortadas , Masculino , Malondialdeído/sangue , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Estresse Oxidativo/efeitos dos fármacos
15.
Int J Mol Sci ; 22(14)2021 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-34298945

RESUMO

Increased oxidative stress is a crucial factor for the progression of cellular senescence and aging. The present study aimed to investigate the effects of licochalcone D (Lico D) on oxidative stress-induced senescence, both in vitro and in vivo, and explore its potential mechanisms. Hydrogen peroxide (200 µM for double time) and D-galactose (D-Gal) (150 mg/kg) were used to induce oxidative stress in human bone marrow-mesenchymal stem cells (hBM-MSCs) and mice, respectively. We performed the SA-ß-gal assay and evaluated the senescence markers, activation of AMPK, and autophagy. Lico D potentially reduced oxidative stress-induced senescence by upregulating AMPK-mediated activation of autophagy in hBM-MSCs. D-Gal treatment significantly increased the expression levels of senescence markers, such as p53 and p21, in the heart and hippocampal tissues, while this effect was reversed in the Lico D-treated animals. Furthermore, a significant increase in AMPK activation was observed in both tissues, while the activation of autophagy was only observed in the heart tissue. Interestingly, we found that Lico D significantly reduced the expression levels of the receptors for advanced glycation end products (RAGE) in the hippocampal tissue. Taken together, our findings highlight the antioxidant, anti-senescent, and cardioprotective effects of Lico D and suggest that the activation of AMPK and autophagy ameliorates the oxidative stress-induced senescence.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Senescência Celular/efeitos dos fármacos , Chalconas/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Envelhecimento/efeitos dos fármacos , Envelhecimento/metabolismo , Animais , Antioxidantes/farmacologia , Autofagia/efeitos dos fármacos , Cardiotônicos/farmacologia , Células Cultivadas , Galactose/metabolismo , Coração/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Humanos , Peróxido de Hidrogênio/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Regulação para Cima/efeitos dos fármacos
16.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201662

RESUMO

Gene expression and phytohormone contents were measured in response to elevating ascorbate in the absence of other confounding stimuli such as high light and abiotic stresses. Young Arabidopsis plants were treated with 25 mM solutions of l-galactose pathway intermediates l-galactose (l-gal) or l-galactono-1,4-lactone (l-galL), as well as L-ascorbic acid (AsA), with 25 mM glucose used as control. Feeding increased rosette AsA 2- to 4-fold but there was little change in AsA biosynthetic gene transcripts. Of the ascorbate recycling genes, only Dehydroascorbate reductase 1 expression was increased. Some known regulatory genes displayed increased expression and included ANAC019, ANAC072, ATHB12, ZAT10 and ZAT12. Investigation of the ANAC019/ANAC072/ATHB12 gene regulatory network revealed a high proportion of ABA regulated genes. Measurement of a subset of jasmonate, ABA, auxin (IAA) and salicylic acid compounds revealed consistent increases in ABA (up to 4.2-fold) and phaseic acid (PA; up to 5-fold), and less consistently certain jasmonates, IAA, but no change in salicylic acid levels. Increased ABA is likely due to increased transcripts for the ABA biosynthetic gene NCED3. There were also smaller increases in transcripts for transcription factors ATHB7, ERD1, and ABF3. These results provide insights into how increasing AsA content can mediate increased abiotic stress tolerance.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Ácido Ascórbico/metabolismo , Glutationa Transferase/genética , Reguladores de Crescimento de Plantas/metabolismo , Estresse Fisiológico/fisiologia , Ácido Abscísico/metabolismo , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Ascorbato Oxidase/genética , Ascorbato Oxidase/metabolismo , Ácido Ascórbico/genética , Ciclopentanos/metabolismo , Galactose/farmacologia , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Glutationa Transferase/metabolismo , Ácidos Hexurônicos/metabolismo , Ácidos Indolacéticos/metabolismo , Oxilipinas/metabolismo , Reguladores de Crescimento de Plantas/genética , Sesquiterpenos/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
17.
Int J Mol Sci ; 22(14)2021 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-34298952

RESUMO

In light of recent research, there is increasing evidence showing that extracellular semen components have a significant impact on the immune reaction of the female partner, leading to the tolerogenic response enabling the embryo development and implantation as well as further progress of healthy pregnancy. Seminal plasma glycoproteins are rich in the unique immunomodulatory glycoepitopes that may serve as ligands for endogenous lectins that decorate the surface of immune cells. Such interaction may be involved in modulation of the maternal immune response. Among immunomodulatory glycans, Lewis type antigens have been of interest for at least two decades, while the importance of T/Tn antigens and related structures is still far from understanding. In the current work, we applied two plant lectins capable of distinguishing glycoepitopes with terminal GalNAc and Gal to identify glycoproteins that are their efficient carriers. By means of lectin blotting and lectin affinity chromatography followed by LC-MS, we identified lactotransferrin, prolactin inducible protein as well as fibronectin and semenogelins 1 and 2 as lectin-reactive. Net-O-glycosylation analysis results indicated that the latter three may actually carry T and/or Tn antigens, while in the case of prolactin inducible protein and lactotransferrin LacdiNAc and lactosamine glycoepitopes were more probable. STRING bioinformatics analysis linked the identified glycoproteins in the close network, indicating their involvement in immune (partially innate) processes. Overall, our research revealed potential seminal plasma ligands for endogenous Gal/GalNAc specific lectins with a possible role in modulation of maternal immune response during fertilization.


Assuntos
Acetilgalactosamina/imunologia , Fertilização/imunologia , Galactose/imunologia , Glicoproteínas/imunologia , Sêmen/imunologia , Proteínas de Plasma Seminal/imunologia , Feminino , Humanos
18.
Molecules ; 26(12)2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-34208735

RESUMO

Saccharides are the most common carbon source for Streptococcus thermophilus, which is a widely used bacterium in the production of fermented dairy products. The performance of the strain is influenced by the consumption of different saccharides during fermentation. Therefore, a precise measurement of the concentrations of saccharides in the fermentation media is essential. An 18-min long method with limits of quantitation in the range of 0.159-0.704 mg/L and with 13C labelled internal standards employing hydrophilic interaction chromatography coupled to mass spectrometric detection-(HILIC-LC-MS) allowed for simultaneous quantification of five saccharides: fructose, glucose, galactose, sucrose, and lactose in the fermentation samples. The method included a four-step sample preparation protocol, which could be easily applied to high-throughput analysis. The developed method was validated and applied to the fermentation samples produced by Streptococcus thermophilus.


Assuntos
Cromatografia Líquida/métodos , Fermentação/fisiologia , Espectrometria de Massas em Tandem/métodos , Dissacarídeos/química , Dissacarídeos/metabolismo , Frutose/metabolismo , Galactose/metabolismo , Glucose/metabolismo , Lactose/metabolismo , Monossacarídeos/química , Monossacarídeos/metabolismo , Streptococcus thermophilus/metabolismo , Sacarose/metabolismo
19.
Molecules ; 26(14)2021 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-34299429

RESUMO

Hybrid bioinorganic biocatalysts have received much attention due to their simple synthesis, high efficiency, and structural features that favor enzyme activity and stability. The present work introduces a biomineralization strategy for the formation of hybrid nanocrystals from ß-galactosidase. The effects of the immobilization conditions were studied, identifying the important effect of metal ions and pH on the immobilization yield and the recovered activity. For a deeper understanding of the biomineralization process, an in silico study was carried out to identify the ion binding sites at the different conditions. The selected ß-galactosidase nanocrystals showed high specific activity (35,000 IU/g biocatalyst) and remarkable thermal stability with a half-life 11 times higher than the soluble enzyme. The nanobiocatalyst was successfully tested for the synthesis of galacto-oligosaccharides, achieving an outstanding performance, showing no signs of diffusional limitations. Thus, a new, simple, biocompatible and inexpensive nanobiocatalyst was produced with high enzyme recovery (82%), exhibiting high specific activity and high stability, with promising industrial applications.


Assuntos
Enzimas Imobilizadas/química , Enzimas/química , beta-Galactosidase/química , Sítios de Ligação/fisiologia , Biomineralização/fisiologia , Simulação por Computador , Estabilidade Enzimática , Enzimas/metabolismo , Enzimas Imobilizadas/metabolismo , Galactose/química , Concentração de Íons de Hidrogênio , Nanopartículas/química , Oligossacarídeos/química , Temperatura , beta-Galactosidase/metabolismo
20.
Int J Mol Sci ; 22(13)2021 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-34202112

RESUMO

D-galactose (D-gal) administration causes oxidative disorder and is widely utilized in aging animal models. Therefore, we subcutaneously injected D-gal at 200 mg/kg BW dose to assess the potential preventive effect of thymoquinone (TQ) and curcumin (Cur) against the oxidative alterations induced by D-gal. Other than the control, vehicle, and D-gal groups, the TQ and Cur treated groups were orally supplemented at 20 mg/kg BW of each alone or combined. TQ and Cur effectively suppressed the oxidative alterations induced by D-gal in brain and heart tissues. The TQ and Cur combination significantly decreased the elevated necrosis in the brain and heart by D-gal. It significantly reduced brain caspase 3, calbindin, and calcium-binding adapter molecule 1 (IBA1), heart caspase 3, and BCL2. Expression of mRNA of the brain and heart TP53, p21, Bax, and CASP-3 were significantly downregulated in the TQ and Cur combination group along with upregulation of BCL2 in comparison with the D-gal group. Data suggested that the TQ and Cur combination is a promising approach in aging prevention.


Assuntos
Benzoquinonas/farmacologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Curcumina/farmacologia , Galactose/farmacologia , Miocárdio/metabolismo , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Benzoquinonas/química , Curcumina/química , Imuno-Histoquímica , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Especificidade de Órgãos , Ratos , Relação Estrutura-Atividade
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