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1.
J Sci Food Agric ; 103(2): 811-819, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36036167

RESUMO

BACKGROUND: Using transglutaminase (TGase) is a new method to improve protein properties in order to promote protein glycosylation. This article mainly studies soy protein isolate (SPI) and glucosamine to improve the freeze-thaw stability of emulsion under the action of TGase. The degree of glycosylation was studied by the content of free amino groups and the degree of conjugation. The optimal conditions for preparing soy protein isolate-glucosamine (SPI-G) conjugate were determined by a response surface optimization model based on single-factor experiments using the creaming index of the emulsion after the first freeze-thaw cycle as the response value. RESULTS: The results showed that the emulsion had the lowest creaming index when the conditions of protein concentration was 20 g L-1 , mass ratio of SPI-G was 5:3 (w/w), enzyme addition amount was 10 U g-1 , and reaction time was 2 h. The optimized modified product was measured for the creaming index after the first freeze-thaw cycle. It was found that the creaming index of the modified product SPI-G after the first freeze-thaw cycle was 9.02%, which was less than and close to the optimized model predicted value. The creaming index and optical microscopy results after three freeze-thaw cycles confirmed that the freeze-thaw stability of the SPI-G samples was significantly enhanced after optimization of the response surface model. CONCLUSION: It showed that glycosylation promoted by TGase could improve the freeze-thaw stability of SPI emulsion, thereby broadening the application of SPI in food. © 2022 Society of Chemical Industry.


Assuntos
Glucosamina , Proteínas de Soja , Proteínas de Soja/química , Emulsões/química , Glucosamina/química , Congelamento , Fenômenos Químicos , Transglutaminases
2.
Carbohydr Polym ; 301(Pt A): 120304, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36436867

RESUMO

Chitin is an essential structural component of complex and dynamic fungal cell walls. It may be converted by partial or full deacetylation to yield chitosan. Here, we describe a method to quantify N-acetyl d-glucosamine (GlcNAc, A) and d-glucosamine (GlcN, D) units and, thus, total amount and average fraction of acetylation (x̅ FA) of the chitinous polymers by complete enzyme hydrolysis of the polymers followed by mass spectrometric analyses of the monomers. First, the native polymers were isotopically N-acetylated, then enzymatically hydrolyzed to A and R (2H3N-acetyl-d-glucosamine - former D) monomers. Relative abundances of A and R units were used to calculate x̅ FA, and a double-isotopically labeled internal standard R* ([13C2,2H3] N-acetyl-d-glucosamine) monomer was used to calculate the absolute amounts of GlcNAc and GlcN units present in the fungal samples. The method was validated using known chitosan polymers and is suitable for both purified cell walls and whole mycelia.


Assuntos
Quitina , Quitosana , Quitina/química , Quitosana/química , Polímeros , Acetilglucosamina , Glucosamina/química , Parede Celular
3.
Nat Commun ; 13(1): 7404, 2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36456551

RESUMO

T cell development requires the coordinated rearrangement of T cell receptor (TCR) gene segments and the expression of either αß or γδ TCR. However, whether and how de novo synthesis of nutrients contributes to thymocyte commitment to either lineage remains unclear. Here, we find that T cell-specific deficiency in glutamine:fructose-6-phosphate aminotransferase 1 (GFAT1), the rate-limiting enzyme of the de novo hexosamine biosynthesis pathway (dn-HBP), attenuates hexosamine levels, blunts N-glycosylation of TCRß chains, reduces surface expression of key developmental receptors, thus impairing αß-T cell ontogeny. GFAT1 deficiency triggers defects in N-glycans, increases the unfolded protein response, and elevates  γδ-T cell numbers despite reducing γδ-TCR diversity. Enhancing TCR expression or PI3K/Akt signaling does not reverse developmental defects. Instead, dietary supplementation with the salvage metabolite, glucosamine, and an α-ketoglutarate analogue partially restores αß-T cell development in GFAT1T-/- mice, while fully rescuing it in ex vivo fetal thymic organ cultures. Thus, dn-HBP fulfils, while salvage nutrients partially satisfy, the elevated demand for hexosamines during early T cell development.


Assuntos
Glucosamina , Hexosaminas , Animais , Camundongos , Fosfatidilinositol 3-Quinases , Nutrientes , Receptores de Antígenos de Linfócitos T gama-delta
4.
Pestic Biochem Physiol ; 188: 105273, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36464378

RESUMO

The chitin metabolic pathway is one of the most lucrative targets for designing pest management regimes. Inhibition of the chitin synthesis pathway causes detrimental effects on the normal growth and development of insects. Phospho-N-acetylglucosamine mutase (AGM) and UDP-N-acetylglucosamine pyrophosphorylase (UAP) are two key chitin biosynthesis enzymes in insects including Helicoverpa armigera, a pest of global significance. In the present study, we have identified, cloned and recombinantly expressed AGM and UAP from H. armigera (HaAGM and HaUAP). Biochemical characterization of recombinant HaAGM and HaUAP exhibited high affinities for their natural substrates N-acetyl glucosamine-6-phosphate (Km 38.72 ± 2.41) and N-acetyl glucosamine-1-phosphate (Km 3.66 ± 0.13), respectively. In the coupled enzyme-catalytic assay, HaAGM and HaUAP yielded the end-products, inorganic pyrophosphate and UDP-GlcNAc, confirming their active participation in the chitin synthesis pathway of H. armigera. Gene expression profiling revealed that HaAGM and HaUAP genes were expressed in all developmental stages and key tissues. These genes also showed substantial responses towards the moulting hormone 20-hydroxyecdysone and chitin biosynthesis inhibitor, novaluron. Remarkably, the RNAi-mediated knockdown of either HaAGM or HaUAP led to severe developmental deformities and significant mortality ranging from 65.61 to 72.54%. Overall findings suggest that HaAGM and HaUAP play crucial roles in the ecdysis and survival of H. armigera. Further, these genes could serve as potential targets for designing pest management strategies for H. armigera.


Assuntos
Muda , Mariposas , Animais , Muda/genética , Quitina , Ecdisterona/farmacologia , Glucosamina , Mariposas/genética
5.
Nanoscale ; 14(45): 17053-17064, 2022 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-36367284

RESUMO

Despite advances in diagnostic and therapeutic methods, the prognosis of patients with hepatocellular carcinoma (HCC) remains poor due to the delay in diagnosis. Herein, we aimed to discover a highly sensitive and specific biomarker for HCC based on genomic big data analysis and create an HCC-targeted imaging probe using carbon nanodots (CNDs) as contrast agents. In genomic analysis, we selected glucose transporter 2 (GLUT2) as a potential imaging target for HCC. We confirmed the target suitability by immunohisto-chemistry tests of 339 patient samples, where 81.1% of the patients exhibited underexpression of GLUT2, i.e., higher GLUT2 intensity in non-tumor tissues than in tumor tissues. To visualize GLUT2, we conjugated CNDs with glucosamine (GLN) as a targeting ligand to yield glucosamine-labeled CNDs (GLN-CNDs). A series of in vitro and in vivo experiments were conducted on GLUT2-modified HepG2 cells to confirm the specificity of the GLN-CNDs. Since the GLUT2 expression is higher in hepatocytes than in HCC cells, the GLUT2-targeted contrast agent is highly attached to normal cells. However, it is possible to produce images in the same form as the images obtained with a cancer cell-targeted contrast agent by inverting color scaling. Our results indicate that GLUT2 is a promising target for HCC and that GLN-CNDs may potentially be used as targeted imaging probes for diagnosing HCC.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carbono , Carcinoma Hepatocelular/diagnóstico por imagem , Meios de Contraste , Neoplasias Hepáticas/diagnóstico por imagem , Glucosamina
6.
Artigo em Russo | MEDLINE | ID: mdl-36440775

RESUMO

The dominant collagen of the cartilaginous matrix in adults is type II collagen. The amount of type II collagen in the intercellular matrix of cartilage is significantly reduced against the background of musculoskeletal system diseases. The basis of articular cartilage is hyaline cartilage tissue consisting of chondrocytes with tissue-specific antigens that induce the production of antibodies in patients with osteoarthritis (OA). Today, new approaches are being considered in the treatment of OA with the use of udenatured type II collagen (UC-II). Such molecular mechanisms of action of UC-II as the formation of a systemic response through oral tolerance are discussed, since the induction of tolerance is the immune pathway, by default, in the intestine. A number of experimental, preclinical (on volunteers) and clinical studies have shown the effectiveness and safety of the use of UC-II in OA. Standardized extracts of UC-II exhibit anti-inflammatory, immunoregulatory, chondroprotective effects, contributing to the reduction of pain symptoms of OA. Against the background of taking UC-II with induced OA, there is a statistically significant decrease in the level of proinflammatory cytokines, such as interleukin (IL-1ß, IL-6), tumor necrosis factor alpha (TNF), C-reactive protein (CRP) in serum and the level of max proteinases (MMP-3), nucleated factor «kappa-bi¼ (NF-κB) in the knee joint. UC-II significantly inhibits the production of prostaglandin E2 (by 20%) and the expression of genes encoding proinflammatory proteins. In experimental models and in OA patients, a decrease in the severity of pain syndrome, an increase in endurance, mobility and an improvement in the functional state of the joints were noted. Clinically, no changes in the structure of the muscle fiber were detected with increased physical exertion. With OA on the background of UC-II (10-40 mg/s), there was a statistically significant decrease in joint pain according to WOMAC. A promising direction of OA therapy is the combination of UC-II with chondroitin sulfate and glucosamine sulfate.


Assuntos
Cartilagem Articular , Dor Musculoesquelética , Osteoartrite , Adulto , Humanos , Colágeno Tipo II/uso terapêutico , Dor Musculoesquelética/tratamento farmacológico , Osteoartrite/tratamento farmacológico , Glucosamina/uso terapêutico , Cartilagem Articular/patologia
7.
Sci Rep ; 12(1): 19813, 2022 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-36396950

RESUMO

The aim of current study was to develop the transdermal transfersomes of glucosamine for better drug delivery. Stretch ability and plasticity of transfersomes membranes mitigate the risk of vesicle rupture in the skin and allows the drug carrying transfersome to pass through the epidermis following the natural water gradient. Transdermal delivery of Glucosamine has an advantage over oral route, having greater local concentration and fewer systemic effects. Thin Film Rotary method was use to prepare transfersomes, and characterization was carried out physio-chemically using electron microscopic studies, zeta potential evaluation, entrapment efficiency studies. To add on in the stability, development of a secondary topical vehicle using Carbopol 940 was carried out to enhance the shelf life of transfersomes. Furthermore, in vivo studies on rabbits were also carried out using the papain induced arthritis model to support the effectiveness of treatment. The radiology studies of knee joint of rabbits proved the effectiveness of glucosamine loaded transfersomes in healing the osteoarthritis with the blood plasma analysis remain unaltered. In vitro characterization showed the successful development of nano-deformable entities with good entrapment efficiency but with little stability, therefore modified into a gel. In a nut shell this modified new dosage from can be best alternative to other conventional options that owe lot of demerits.


Assuntos
Artrite , Absorção Cutânea , Animais , Coelhos , Papaína , Glucosamina , Administração Cutânea
8.
Int J Mol Sci ; 23(19)2022 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-36232791

RESUMO

Jellyfishes are considered a new potential resource in food, pharmaceutical and biomedical industries. In these latter cases, they are studied as source of active principles but are also exploited to produce marine collagen. In the present work, jellyfish skin polysaccharides (JSP) with glycosaminoglycan (GAG) features were extracted from Rhizostoma pulmo, a main blooming species of Mediterranean Sea, massively augmented by climate leaded "jellyfishication" of the sea. Two main fractions of R. pulmo JSP (RP-JSPs) were isolated and characterized, namely a neutral fraction (RP-JSP1) and a sulphate rich, negatively charged fraction (RP-JSP2). The two fractions have average molecular weights of 121 kDa and 590 kDa, respectively. Their sugar composition was evaluated through LC-MS analysis and the result confirmed the presence of typical GAG saccharides, such as glucose, galactose, glucosamine and galactosamine. Their use as promoters of wound healing was evaluated through in vitro scratch assay on murine fibroblast cell line (BALB/3T3 clone A31) and human keratinocytes (HaCaT). Both RP-JSPs demonstrated an effective confluency rate activity leading to 80% of scratch repair in two days, promoting both cell migration and proliferation. Additionally, RP-JSPs exerted a substantial protection from oxidative stress, resulting in improved viability of treated fibroblasts exposed to H2O2. The isolated GAG-like polysaccharides appear promising as functional component for biomedical skin treatments, as well as for future exploitation as pharmaceutical excipients.


Assuntos
Cnidários , Cifozoários , Animais , Carboidratos , Colágeno/metabolismo , Excipientes , Fibroblastos/metabolismo , Galactosamina , Galactose , Glucosamina , Glucose , Glicosaminoglicanos , Humanos , Peróxido de Hidrogênio , Camundongos , Polissacarídeos/farmacologia , Cifozoários/metabolismo , Sulfatos , Cicatrização
9.
Carbohydr Polym ; 298: 120131, 2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36241332

RESUMO

With the purpose of investigating the bio-interaction between CNC and cells, the end-fluorescence CNC (FCNC) and surface-adsorbed glucosamine [(g)FCNC] were prepared by the region-selective modification and electrostatic adsorption strategy in this study. The cell growth was arrested in a time dependent manner when incubated with CNC in the low glucose environment. The small-size FCNC was preferred by the cells with the observation of higher affinity. Specifically, the affinity of (g)FCNC to the phagocytic cell RAW264.7 cell and kidney derived cell 293 t was generally increased in response to the "Warburg effect", which on the contrary was observed with the weak effect in the tumor cells. We confirmed the exocytosis of internalized rod-like nanocrystal was achieved by the packaging effect of exosomes. Our results revealed the underappreciated bio-interaction between CNC and different cells, which contributed the evidence of an inspiration for this natural nanomaterial in the cell-level application.


Assuntos
Exossomos , Nanopartículas , Celulose/química , Celulose/farmacologia , Exocitose , Glucosamina , Glucose , Nanopartículas/química
10.
Chem Commun (Camb) ; 58(89): 12479-12482, 2022 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-36278312

RESUMO

In the present study, we synthesized a novel near-infrared turn-on BODIPY probe and a new norbornene-modified glucosamine derivative. The probe exhibits a significant NIR fluorescence emission with a turn-on response and can perform tumour-specific imaging in tumour-bearing mice. The non-natural glucosamine provides metabolic glycoengineering labelling. It can be expressed on cells as chemical tags and further reacted with fluorescence dyes for cell labelling. The combination of the two derivatives enables quick and sensitive cell imaging in vitro and in vivo using the iEDDA reaction.


Assuntos
Química Click , Neoplasias , Camundongos , Animais , Glucosamina , Imagem Óptica , Corantes Fluorescentes , Norbornanos
11.
Sci Rep ; 12(1): 17834, 2022 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-36284223

RESUMO

Soil microorganisms are critical for soil carbon (C) cycling. They primarily regulate the turnover of the soil organic C (SOC) by adjusting their community structure, and contributing residues with a considerable amount to the resistant SOC. Nevertheless, how long-term fertilization (e.g., the combination of manure and chemical fertilizer) affects the spatial distribution of both living microbial communities and dead microbial residue within soil aggregate fractions remains largely unclear. In this study, we analyzed changes in microbial community (lipid biomarkers) and microbial residue retention (amino sugar biomarkers), and also calculated the contribution of microbial residue to organic C in bulk soil and different soil aggregates (> 2 mm, 1-2 mm, 0.25-1 mm, and < 0.25 mm) in Alfisols treated with 29 years fertilization or no fertilization (control). Our results showed that long-term fertilization significantly increased the mean weight diameter (MWD) of aggregates and organic C contents in all aggregate fractions. The fertilization treatment increased the contents of PLFAs and microbial residue C, but the relative contribution of microbial residue to SOC was higher in the control (56.8% vs. 49.0%), due to the low SOC background caused by much lower level of non-microbially derived C input. These results suggested that long-term fertilization could increase SOC by accumulating both plant- and microbial-derived C, while the C deficient soil is more dependent on the accumulation of microbial residues. Long-term fertilization promoted the enrichment of bacterial-derived muramic acid in micro aggregates, but increased the proportion of fungal-derived glucosamine in macro aggregates. Meanwhile, the contribution of bacterial residue to organic C in the fertilization treatment was higher in micro aggregates (7.6% for > 2 mm vs. 9.2% for < 0.25 mm aggregate), while the contribution of fungal residue was higher in macro aggregate fractions (40.9% for > 2 mm vs. 35.7% for < 0.25 mm aggregate). The above results indicated that long-term fertilization could drive the differentiation of heterogeneous microbial residue accumulation patterns that significantly alter the contribution of fungal- versus bacterial-derived C to organic C within soil aggregate fractions.


Assuntos
Fertilizantes , Solo , Solo/química , Fertilizantes/análise , Esterco , Ácidos Murâmicos , Carbono , Microbiologia do Solo , Bactérias , Amino Açúcares , Glucosamina , Lipídeos
12.
Mar Drugs ; 20(10)2022 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-36286469

RESUMO

Excessive inorganic ions in vivo may lead to electrolyte disorders and induce damage to the human body. Therefore, preparation of enhanced bioactivity compounds, composed of activated organic cations and organic anions, is of great interest among researchers. In this work, glucosamine-heparin salt (GHS) was primarily synthesized with positively charged glucosamine hydrochloride (GAH) and negatively charged heparin sodium (Heps) by ion exchange method. Then, the detailed structural information of the GHS was characterized by FTIR, 1H NMR spectroscopy and ICP-MS. In addition, its anticoagulant potency and antioxidant properties were evaluated, respectively. The results demonstrated that GHS salt achieved enhanced antioxidant activities, including 98.78% of O2•- radical scavenging activity, 91.23% of •OH radical scavenging rate and 66.49% of DPPH radical scavenging capacity at 1.6 mg/mL, severally. Meanwhile, anticoagulant potency (ATTP) of GHS strengthened from 153.10 ± 17.14 to 180.03 ± 6.02 at 0.75 µmol/L. Thus, introducing cationic glucosamine residues into GHS could improve its anticoagulant activity. The findings suggest that GHS product with a small amount of inorganic ions can greatly abate the prime cost of antioxidants and anticoagulants, and has significant economic benefits and practical significance.


Assuntos
Anticoagulantes , Heparina , Humanos , Heparina/farmacologia , Heparina/química , Anticoagulantes/farmacologia , Anticoagulantes/química , Antioxidantes/farmacologia , Antioxidantes/química , Glucosamina/farmacologia , Glucosamina/química , Cloreto de Sódio , Íons , Eletrólitos
13.
Int J Mol Sci ; 23(20)2022 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-36293092

RESUMO

The Gram-positive bacterium Staphylococcus aureus is responsible for serious acute and chronic infections worldwide and is well-known for its biofilm formation ability. Recent findings of biofilms on dry hospital surfaces emphasise the failures in current cleaning practices and disinfection and the difficulty in removing these dry surface biofilms (DSBs). Many aspects of the formation of complex DSB biology on environmental surfaces in healthcare settings remains limited. In the present study, we aimed to determine how the protein component varied between DSBs and traditional hydrated biofilm. To do this, biofilms were grown in tryptic soy broth (TSB) on removable polycarbonate coupons in the CDC biofilm reactor over 12 days. Hydrated biofilm (50% TSB for 48 h, the media was then changed every 48 h with 20% TSB, at 37 °C with 130 rpm). DSB biofilm was produced in 5% TSB for 48 h at 35 °C followed by extended periods of dehydration (48, 66, 42 and 66 h at room temperature) interspersed with 6 h of 5% TSB at 35 °C. Then, we constructed a comprehensive reference map of 12-day DSB and 12-day hydrated biofilm associated proteins of S. aureus using a high-throughput tandem mass tag (TMT)-based mass spectrometry. Further pathway analysis of significantly differentially expressed identified proteins revealed that proteins significantly upregulated in 12-day DSB include PTS glucose transporter subunit IIBC (PtaA), UDP-N-acetylmuramate-L-alanine ligase (MurC) and UDP-N-acetylenolpyruvoylglucosamine (MurB) compared to 12-day hydrated biofilm. These three proteins are all linked with peptidoglycan biosynthesis pathway and are responsible for cell-wall formation and thicker EPS matrix deposition. Increased cell-wall formation may contribute to the persistence of DSB on dry surfaces. In contrast, proteins associated with energy metabolisms such as phosphoribosyl transferase (PyrR), glucosamine--fructose-6-phosphate aminotransferase (GlmS), galactose-6-phosphate isomerase (LacA), and argininosuccinate synthase (ArgG) were significantly upregulated whereas ribosomal and ABC transporters were significantly downregulated in the 12-day hydrated biofilm compared to DSB. However, validation by qPCR analysis showed that the levels of gene expression identified were only partially in line with our TMT-MS quantitation analysis. For the first time, a TMT-based proteomics study with DSB has shed novel insights and provided a basis for the identification and study of significant pathways vital for biofilm biology in this reference microorganism.


Assuntos
Infecções Estafilocócicas , Staphylococcus aureus , Humanos , Proteômica , Argininossuccinato Sintase , Quebras de DNA de Cadeia Dupla , Peptidoglicano , Biofilmes , Glucosamina , Transferases , Transportadores de Cassetes de Ligação de ATP , Proteínas Facilitadoras de Transporte de Glucose , Transaminases , Alanina , Difosfato de Uridina
14.
Chemosphere ; 309(Pt 2): 136724, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36208803

RESUMO

Antibiotics are detected worldwide in the aquatic environment, with continuously rising concentrations. Antibiotics in the environment have the potential to damage ecosystems and contribute to the development of resistance. Whilst a few antibiotics, such as some ß-lactams, are eliminated by effluent treatment, others, such as fluoroquinolones, are not or just partially removed and enter the environment. Therefore, approaches are needed to tackle those problems at the compound level. Benign by design (BbD), an important part of green pharmacy, has the goal to integrate environmental fate and end-of-use considerations at the very beginning, i.e., into the design of active pharmaceutical ingredients. Hence, pharmaceuticals should be designed to be sufficiently active and stable during storage and usage but should degrade after excretion into the environment, so that they cannot cause any adverse effects. Fluoroquinolones (FQs) are important broad-spectrum antibiotics. They are known to be persistent in the environment and to be neither inactivated nor degraded or even mineralized during sewage treatment. The addition of new substituents via amidation, like glucosamine moieties, at the carboxylic group of FQs, led to better antimicrobial activity compared to its parent compounds against various microorganisms. To investigate if the addition of sugar moieties could improve the overall environmental biodegradability of FQs, eight novel quinolone and fluoroquinolone analogs conjugated with 1,3,4,6-Tetra-O-acetyl-ß-d-glucosamine and 2-deoxy-d-glucopyranose have been investigated regarding their ready biodegradability (OECD 301D/F) and their degradation pathways have been analyzed. According to the OECD 301D test, none of the substances could be classified as readily biodegradable. However, the O-acetyl analogs did undergo a partial degradation of the O-acetyl glucosamine moiety, via stepwise deacetylation and the degradation of the whole glucosamine moiety. The degradation resulted in Fluoroquinolone-3-carboxamide derivatives. Those insights could be further used as input for fragment-based design of benign APIs that will degrade once they reached the environment.


Assuntos
Fluoroquinolonas , Quinolonas , Fluoroquinolonas/toxicidade , Glucosamina , Esgotos , Ecossistema , Antibacterianos/farmacologia , Antibacterianos/metabolismo , beta-Lactamas , Açúcares , Preparações Farmacêuticas
15.
Nanoscale ; 14(44): 16467-16478, 2022 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-36305892

RESUMO

The study of biologically relevant molecules and their interaction with external stimuli on a single molecular scale is of high importance due to the availability of distributed rather than averaged information. Surface enhanced Raman scattering (SERS) provides direct chemical information, but is rather challenging on the single molecule (SM) level, where it is often assumed to require a direct contact of analyte molecules with the metal surface. Here, we detect and investigate the molecular states of single hemin by SM-SERS. A DNA aptamer based G-quadruplex mediated recognition of hemin directs its placement in the SERS hot-spot of a DNA Origami Nanofork Antenna (DONA). The configuration of the DONA structure allows the molecule to be trapped at the plasmonic hot-spot preferentially in no-contact configuration with the metal surface. Owing to high field enhancement at the plasmonic hot spot, the detection of a single folded G-quadruplex becomes possible. For the first time, we present a systematic study by SM-SERS where most hemin molecule adopt a high spin and oxidation state (III) that showed state crossover to low spin upon strong-field-ligand binding. The present study therefore, provides a platform for studying biologically relevant molecules and their properties at SM sensitivity along with demonstrating a conceptual advancement towards successful monitoring of single molecular chemical interaction using DNA aptamers.


Assuntos
Nanopartículas Metálicas , Análise Espectral Raman , Ouro/química , Hemina , Nanopartículas Metálicas/química , DNA/química , Glucosamina
16.
Arch Microbiol ; 204(11): 659, 2022 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-36190580

RESUMO

Two strains (TRM95111T and TRM95001T) of Gram-stain-negative, aerobic, rod-shaped microbes were isolated from the nodule and rhizosphere of Lotus japonicus grown in the campus of Tarim University in Alar, Xinjiang, China. Strain TRM95111T and strain TRM95001T shared 93.1% 16S rRNA gene sequences similarity with each other and had 98.2 and 97.9% 16S rRNA gene sequence similarity to the closest species Rhizobium subbaraois JC85T and R. halotolerans AB21T by EzBioCloud blast, respectively. Phylogenetic analyses based on 16S rRNA gene sequences, housekeeping gene sequences and core-proteome average amino acid identity (cpAAI) showed that two strains belonged to the genus Rhizobium. The value of digital DNA-DNA hybridization (dDDH) between strain TRM95111T and the closest strain R. subbaraonis JC85T was 21.8%, respectively. The dDDH value between strain TRM95001T and the closest strains R. tarimense PL-41T was 27.1%. Whole-genome average nucleotide identity (ANI) values of the strain TRM95111T were 75.6-79.3% and strain TRM95001T were 79.2-83.8%, compared to their closely related strains. The G + C content values of strain TRM95111T and TRM95001T were 65.1 and 60.7 mol%, respectively. Two isolates contained predominant quinone of Q-10 and the major fatty acids was C18:1ω7c and they were sensitive to 1 µg of amikacin and kanamycin. The polar lipids of strain TRM95111T included unidentified aminophospho lipids (APL1-3), unidentified phospholipids (PL1-2), phosphatidylcholine (PC), unidentified lipids (L1-5) and phospholipids of unknown structure containing glucosamine (NPG), compared to the polar lipids of strain TRM95001T including unidentified aminophospho lipids (APL1-3), unidentified phospholipids (PL1-2), phosphatidylcholine (PC), unidentified lipids (L2-5), hydroxy phosphatidyl ethanolamine (OH-PE) and phospholipids of unknown structure containing glucosamine (NPG). Nodulation tests showed that two strains could induce nodules formation in L. japonicus. Based on the genomic, phenotypic and phylogenetic analyses, strain TRM95111T and strain TRM95001T are suggested to represent two new species of the genus Rhizobium, whose names are proposed as Rhizobium alarense sp. nov. and Rhizobium halophilum sp. nov. The type strains are TRM95111T (=CCTCC AB 2021116T =JCM34826T) and TRM950011T (=CCTCC AB 2021095T =JCM34967T), respectively.


Assuntos
Lotus , Rhizobium , Amicacina , Aminoácidos , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Etanolaminas , Ácidos Graxos/análise , Glucosamina , Humanos , Lotus/genética , Nucleotídeos , Fosfatidilcolinas , Fosfolipídeos/análise , Filogenia , Proteoma/genética , Quinonas , RNA Ribossômico 16S/genética , Rizosfera , Análise de Sequência de DNA
17.
Comput Math Methods Med ; 2022: 6078254, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36081430

RESUMO

Background: Osteoporosis (OP) associated with knee osteoarthritis (KOA) is common in older men and postmenopausal women, and it is important to find reliable and effective treatments for this disease to improve joint function and bone metabolism in this population. Objective: To clarify the clinical efficacy of glucosamine (GlcN) plus sodium hyaluronate (SH) for OP complicated by KOA (OP + KOA) and its influence on joint function and bone metabolic markers (BMMs). Methods: Admitted from July 2019 to July 2021, 126 patients with OP + KOA were selected, including 76 cases (observation group) treated with GlcN plus SH and 50 cases (control group) given GlcN alone. The pain, joint function, BMMs, and clinical efficacy were evaluated and compared. Pain and joint function assessments employed the Visual Analogue Scale (VAS) and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) plus Lysholm Knee Scoring Scale, respectively. BMMs mainly measured bone gla protein (BGP), serum tartrate-resistant acid phosphatase variant (TRACP)-5b, type I collagen cross-linked C-telopeptide (CTX-1), and bone-specific alkaline phosphatase (BALP). Results: Higher posttreatment VAS scores were determined in observation group as compared to control group; observation group showed lower WOMAC scores of joint function and higher Lysholm scores than control group; in terms of BMMs, TRACP-5b and CTX-1 were lower while BGP and BALP were higher in observation group; the curative effect was also higher in observation group. All the above differences were statistically significant. Conclusions: GlcN plus SH has definite clinical efficacy in the treatment of OP + KOA, which can not only significantly improve patients' joint function and bone metabolism but also relieve pain, with high clinical popularization value.


Assuntos
Osteoartrite do Joelho , Osteoporose , Idoso , Feminino , Glucosamina/uso terapêutico , Humanos , Ácido Hialurônico/uso terapêutico , Articulação do Joelho , Masculino , Osteoartrite do Joelho/tratamento farmacológico , Osteoporose/tratamento farmacológico , Dor , Fosfatase Ácida Resistente a Tartarato , Resultado do Tratamento
18.
J Immunol ; 209(9): 1674-1690, 2022 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-36150727

RESUMO

Immunomodulatory (IM) metabolic reprogramming in macrophages (Mϕs) is fundamental to immune function. However, limited information is available for human Mϕs, particularly in response plasticity, which is critical to understanding the variable efficacy of immunotherapies in cancer patients. We carried out an in-depth analysis by combining multiplex stable isotope-resolved metabolomics with reversed phase protein array to map the dynamic changes of the IM metabolic network and key protein regulators in four human donors' Mϕs in response to differential polarization and M1 repolarizer ß-glucan (whole glucan particles [WGPs]). These responses were compared with those of WGP-treated ex vivo organotypic tissue cultures (OTCs) of human non-small cell lung cancer. We found consistently enhanced tryptophan catabolism with blocked NAD+ and UTP synthesis in M1-type Mϕs (M1-Mϕs), which was associated with immune activation evidenced by increased release of IL-1ß/CXCL10/IFN-γ/TNF-α and reduced phagocytosis. In M2a-Mϕs, WGP treatment of M2a-Mϕs robustly increased glucose utilization via the glycolysis/oxidative branch of the pentose phosphate pathway while enhancing UDP-N-acetyl-glucosamine turnover and glutamine-fueled gluconeogenesis, which was accompanied by the release of proinflammatory IL-1ß/TNF-α to above M1-Mϕ's levels, anti-inflammatory IL-10 to above M2a-Mϕ's levels, and attenuated phagocytosis. These IM metabolic responses could underlie the opposing effects of WGP, i.e., reverting M2- to M1-type immune functions but also boosting anti-inflammation. Variable reprogrammed Krebs cycle and glutamine-fueled synthesis of UTP in WGP-treated OTCs of human non-small cell lung cancer were observed, reflecting variable M1 repolarization of tumor-associated Mϕs. This was supported by correlation with IL-1ß/TNF-α release and compromised tumor status, making patient-derived OTCs unique models for studying variable immunotherapeutic efficacy in cancer patients.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , beta-Glucanas , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Glucosamina/metabolismo , Glucose/metabolismo , Glutamina/metabolismo , Humanos , Interleucina-10 , Neoplasias Pulmonares/metabolismo , Macrófagos , NAD/metabolismo , Fagocitose , Triptofano/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Difosfato de Uridina/metabolismo , Uridina Trifosfato/metabolismo , beta-Glucanas/metabolismo
19.
Int J Mol Sci ; 23(18)2022 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-36142319

RESUMO

With osteoarthritis being the most common degenerative disease in pet animals, a very broad panel of natural health products is available on the market for its management. The aim of this systematic review and meta-analysis, registered on PROSPERO (CRD42021279368), was to test for the evidence of clinical analgesia efficacy of fortified foods and nutraceuticals administered in dogs and cats affected by osteoarthritis. In four electronic bibliographic databases, 1578 publications were retrieved plus 20 additional publications from internal sources. Fifty-seven articles were included, comprising 72 trials divided into nine different categories of natural health compound. The efficacy assessment, associated to the level of quality of each trial, presented an evident clinical analgesic efficacy for omega-3-enriched diets, omega-3 supplements and cannabidiol (to a lesser degree). Our analyses showed a weak efficacy of collagen and a very marked non-effect of chondroitin-glucosamine nutraceuticals, which leads us to recommend that the latter products should no longer be recommended for pain management in canine and feline osteoarthritis.


Assuntos
Produtos Biológicos , Canabidiol , Doenças do Gato , Doenças do Cão , Osteoartrite , Animais , Produtos Biológicos/uso terapêutico , Canabidiol/uso terapêutico , Gatos , Condroitina/uso terapêutico , Colágeno/uso terapêutico , Suplementos Nutricionais , Doenças do Cão/tratamento farmacológico , Cães , Glucosamina/uso terapêutico , Osteoartrite/tratamento farmacológico , Osteoartrite/veterinária
20.
J Pak Med Assoc ; 72(7): 1272-1277, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-36156542

RESUMO

OBJECTIVE: To determine the added benefits of short-term glucosamine and chondroitin sulfate supplementation in combination with manual therapy and resistance exercise training in the management of knee osteoarthritis. METHODS: A parallel-design, double-blind randomised controlled trial was conducted from January to September 2020 at the Foundation University Institute of Rehabilitation Sciences and Fauji Foundation Hospital, Rawalpindi, Pakistan, and comprised of knee osteoarthritis patients of either gender having radiological evidence of grade III or less on Kellgren classification. The subjects were randomly allocated to active comparator group A and experimental group B. Both the groups received manual therapy and resistance exercise training, while group B additionally received glucosamine and chondroitin sulfate supplementation for 4 weeks. Study outcomes included pain, function, quality of life, range of motion, strength, fall risk, skeletal muscle mass, visceral fat area, body fat, intracellular water ratio, and segmental lean and fat mass. Data was analysed using SPSS 21. RESULTS: Of the 24 subjects, there were 12(50%) in each of the two groups. Each groups had 9(75%) males and 3(25%) females. In terms knee osteoarthritis grade, there was no significant difference between the groups (p=1.00). No significant differences were observed in any of the outcome measures neither at 2 weeks, nor at 4 weeks post-intervention between the groups (p>0.05) except for percentage change in segmental lean mass of the right leg at 2nd week and of the left leg at 4th week (p<0.05). CONCLUSIONS: Manual therapy and resistance exercise training are effective in the management of knee osteoarthritis, however, glucosamine and chondroitin sulfate supplementation for 4 weeks showed no additional benefits. Clinical Trial Number: NCT04654871. https://www.clinicaltrials.gov/ct2/show/NCT04654871.


Assuntos
Manipulações Musculoesqueléticas , Osteoartrite do Joelho , Treinamento de Força , Água Corporal , Sulfatos de Condroitina/uso terapêutico , Suplementos Nutricionais , Terapia por Exercício , Feminino , Glucosamina/uso terapêutico , Humanos , Masculino , Osteoartrite do Joelho/tratamento farmacológico , Qualidade de Vida , Resultado do Tratamento
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