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1.
Carbohydr Res ; 545: 109256, 2024 Sep 03.
Artículo en Inglés | MEDLINE | ID: mdl-39241666

RESUMEN

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related deaths. Saliva diagnosis is an essential approach for clinical applications owing to its noninvasive and material-rich features. The purpose of this study was to investigate differences in wheat germ agglutinin (WGA)-based recognition of salivary protein N-linked glycan profiles to distinguish non-small cell lung cancer (NSCLC) patients from controls. We used WGA-magnetic particle conjugates to isolate glycoproteins in the pooled saliva of healthy volunteers (HV, n = 35), patients with benign pulmonary disease (BPD, n = 35), lung adenocarcinoma (ADC, n = 35), and squamous cell carcinoma (SCC, n = 35), following to release the N-linked glycans from the isolated proteins with PNGase F, and further identified and annotated the released glycans by MALDI-TOF/TOF-MS, respectively. The results showed that 34, 35, 39, and 44 N-glycans recognized by WGA were identified and annotated from pooled saliva samples of HV, BPD, ADC, and SCC, respectively. Furthermore, the proportion of N-glycans recognized by WGA in BPD (81.2 %), ADC (90.1 %), and SCC (88.7 %), increased compared to HV (71.9 %). Two N-glycan peaks (m/z 2286.799, and 3399.211) specifically recognized by WGA were present only in NSCLC. These findings suggest that altered salivary glycopatterns such as sialic acids and GlcNAc containing N-glycans recognized by WGA might serve as potential personalized biomarkers for the diagnosis of NSCLC patients.

2.
Int J Mol Sci ; 25(14)2024 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-39063110

RESUMEN

Current treatments for lymphoma are plagued by substantial toxicity and the inability to overcome drug resistance, leading to eventual relapse and rationalizing the development of novel, less toxic therapeutics and drug combinations. Histone deacetylase inhibitors (HDACis) are a broad class of epigenetic modulators that have been studied in multiple tumor types, including lymphoma. Currently, HDACis are FDA-approved for treating relapsed T-cell lymphomas and multiple myeloma, with ongoing trials in other lymphomas and solid tumors. As single agents, HDACis frequently elicit toxic side effects and have limited efficacy; therefore, many current treatment strategies focus on combinations to boost efficacy while attempting to minimize toxicity. Fermented wheat germ extract (FWGE) is a complementary agent that has shown efficacy in several malignancies, including lymphoma. Here, we utilize a more potent FWGE derivative, known as fermented wheat germ protein (FWGP), in combination with the HDACi AR42, to assess for enhanced activity. We report increased in vitro killing, cell cycle arrest, and in vivo efficacy for this combination compared to each agent alone with minimal toxicity, suggesting a potentially new, minimally toxic treatment modality for lymphoma.


Asunto(s)
Inhibidores de Histona Desacetilasas , Linfoma , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Animales , Línea Celular Tumoral , Ratones , Linfoma/tratamiento farmacológico , Linfoma/patología , Linfoma/metabolismo , Fermentación , Ensayos Antitumor por Modelo de Xenoinjerto , Triticum/química , Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Proteínas de Plantas/farmacología , Apoptosis/efectos de los fármacos , Aminopiridinas , Benzamidas , Extractos Vegetales
3.
Nan Fang Yi Ke Da Xue Xue Bao ; 44(6): 1024-1032, 2024 Jun 20.
Artículo en Chino | MEDLINE | ID: mdl-38977331

RESUMEN

OBJECTIVE: To investigate the mechanism of 2, 6-dimethoxy-1, 4-benzoquinone (DMQ), an active ingredients in fermented wheat germ extract, for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice. METHODS: Cultured murine bone marrow-derived macrophages (BMDM) stimulated with lipopolysaccharide (LPS) were treated with DMQ, followed by treatment with Nigericin, ATP, and MSU for activating the canonical NLRP3 inflammasome; the noncanonical NLRP3 inflammasome was activated by intracellular transfection of LPS, and AIM2 inflammasome was activated using Poly A: T.In human monocytic THP-1 cells, the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ, the levels of IL-1ß and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA, and the survival time of the mice within 36 h was observed. RESULTS: Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM, but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock, DMQ treatment significantly reduced the levels of IL-1ß in the serum and peritoneal fluid and obviously prolonged survival time of the mice. CONCLUSION: DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPSinduced septic shock in mice.


Asunto(s)
Benzoquinonas , Inflamasomas , Interleucina-1beta , Lipopolisacáridos , Ratones Endogámicos C57BL , Proteína con Dominio Pirina 3 de la Familia NLR , Choque Séptico , Animales , Choque Séptico/tratamiento farmacológico , Choque Séptico/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Ratones , Inflamasomas/metabolismo , Masculino , Humanos , Benzoquinonas/farmacología , Benzoquinonas/uso terapéutico , Interleucina-1beta/metabolismo , Macrófagos/metabolismo , Macrófagos/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo , Células THP-1 , Modelos Animales de Enfermedad
4.
Methods Mol Biol ; 2824: 105-120, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39039409

RESUMEN

The Rift Valley fever virus is one of the bunyaviruses on the WHO's priority list of pathogens that may cause future pandemics. A better understanding of disease progression and viral pathogenesis is urgently needed to develop treatments. The non-structural proteins NSs and NSm of human pathogenic bunyaviruses represent promising therapeutic targets, as they are often key virulence factors. However, their function is still poorly understood, and their structure is yet unknown, mainly because no successful production of these highly complex proteins has been reported. Here we propose a powerful combination of wheat germ cell-free protein synthesis and NMR to study the structure of these proteins and in particular detail cell-free synthesis and lipid reconstitution methods that can be applied to complex membrane proteins.


Asunto(s)
Sistema Libre de Células , Humanos , Espectroscopía de Resonancia Magnética/métodos , Proteínas no Estructurales Virales/metabolismo , Proteínas no Estructurales Virales/química , Triticum/metabolismo , Resonancia Magnética Nuclear Biomolecular/métodos , Biosíntesis de Proteínas , Virus de la Fiebre del Valle del Rift , Proteínas Virales/metabolismo , Proteínas Virales/química
5.
Foods ; 13(12)2024 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-38928894

RESUMEN

Its high dietary fiber and protein contents and nutritional quality make defatted wheat germ (DWG) a valuable cereal by-product, yet its negative impact on food structure limits its use as a food ingredient. In this research, DWG underwent air classification, which identified two fractions with high fiber (HF) and low fiber/high protein (LF) contents, and a bioprocessing protocol, involving treatment with xylanase and fermentation with selected lactic acid bacterial strains. The degree of proteolysis was evaluated through electrophoretic and chromatographic techniques, revealing differences among fractions and bioprocessing options. Fermentation led to a significant increase in free amino acids (up to 6 g/kg), further enhanced by the combination with xylanase. When HF was used as an ingredient in bread making, the fiber content of the resulting bread exceeded 3.6 g/100 g, thus reaching the threshold required to make a "source of fiber" claim according to Regulation EC No.1924/2006. Meanwhile, all breads could be labeled a "source of protein" since up to 13% of the energy was provided by proteins. Overall, bioprocessed ingredients lowered the glycemic index (84 vs. 89) and increased protein digestibility (80 vs. 63%) compared to control breads. Technological and sensory analysis showed that the enzymatic treatment combined with fermentation also conferred a darker and more pleasant color to the bread crust, as well as better crumb porosity and elasticity.

6.
Food Chem ; 457: 140129, 2024 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-38908242

RESUMEN

In this study, chlorogenic acid (CA), piceatannol (PIC), epigallocatechin-3-gallate (EGCG) and ferulic acid (FA) was selected to explore the influence of polyphenol on the structural properties of wheat germ albumin (WGA) and wheat germ globulin (WGG). The emulsifying properties of the emulsions prepared by WGA-EGCG complex were also evaluated. The results indicated that all polyphenols could significantly enhance the antioxidant capacity of WGA and WGG. In particular, EGCG increased the ratio of random coil in WGA and WGG, resulting in protein unfolding and shifting from an order to disorder structure. In addition, lipid oxidation and protein oxidation of the soybean oil emulsion was significantly slowed down by WGA-EGCG. The stability of the emulsions under various environmental stress and the storage time was significantly improved by WGA-EGCG. These findings can provide a reference for expanding the application of wheat germ protein in food industry.


Asunto(s)
Emulsiones , Globulinas , Polifenoles , Triticum , Triticum/química , Polifenoles/química , Polifenoles/farmacología , Globulinas/química , Emulsiones/química , Albúminas/química , Conformación Proteica , Proteínas de Plantas/química , Antioxidantes/química , Antioxidantes/farmacología
7.
J Microsc ; 2024 Jun 17.
Artículo en Inglés | MEDLINE | ID: mdl-38881512

RESUMEN

Breast cancer is one of the leading causes of mortality among women. The tumour microenvironment, consisting of host cells and extracellular matrix, has been increasingly studied for its interplay with cancer cells, and the resulting effect on tumour progression. While the breast is one of the most innervated organs in the body, the role of neurons, and specifically sensory neurons, has been understudied, mostly for technical reasons. One of the reasons is the anatomy of sensory neurons: sensory neuron somas are located in the spine, and their axons can extend longer than a meter across the body to provide innervation in the breast. Next, neurons are challenging to culture, and there are no cell lines adequately representing the diversity of sensory neurons. Finally, sensory neurons are responsible for transporting several different types of signals to the brain, and there are many different subtypes of sensory neurons. The subtypes of sensory neurons, which innervate and interact with breast tumours, are unknown. To establish the tools for labelling and subtyping neurons that interact with breast cancer cells, we utilised two retrograde tracer's standards in neuroscience, wheat-germ agglutinin (WGA) and cholera toxin subunit B (CTB). In vitro, we employed primary sensory neurons isolated from mouse dorsal root ganglia, cultured in a custom-built microfluidic device DACIT, that mimics the anatomical compartmentalisation of the sensory neuron's soma and axons. In vivo, we utilised both syngeneic and transgenic mouse models of mammary carcinoma. We show that CTB and WGA trace different but overlapping sensory neuronal subpopulations: while WGA is more efficient in labelling CGRP+ neurons, CTB is superior in labelling the NF200+ neurons. Surprisingly, both tracers are also taken up by a significant population of breast cancer cells, both in vitro and in vivo. In summary, we have established methodologies for retrograde tracing of sensory neurons interacting with breast cancer cells. Our tools will be useful for future studies of breast tumour innervation, and development of therapies targeting breast cancer-associated neuron subpopulations of sensory neurons. Lay description: Breast cancer is an aggressive disease that affects both women and men throughout the world. While it has been reported that the increasing size of nerves in breast cancer correlates to bad prognosis in patients, the role of nerves, especially sensory nerves, in breast cancer progression, has remained largely understudied. Sensory nerves are responsible for delivering signals such as pain, mechanical forces (pressure, tension, stretch, touch) and temperature to the brain. The human body is densely innervated, and nerves extending into peripheral organs can be as long as a few meters. Nerve classification and function can be very complex, as they contain bundles of extensions (axons) originating in different neuronal bodies (soma). Maintaining neurons and growing axons in cell culture conditions in order to mimic innervation is technically challenging, as it involves multiple organs of the human body. Here, we focus on tracing sensory axons from the breast tumours back to the neuronal soma, located in the dorsal root ganglia, inside the spine. To do so, we are using two different 'retrograde' tracers, WGA and CTB, which are proteins with a natural ability to enter axons and travel in a retrograde fashion, arriving at the soma, even if it means to travel distances longer than a meter. Both tracers are fluorescently labelled, making them visible using high-resolution fluorescent microscopy. We show that both WGA and CTB can label sensory neurons in tumours, or in cell culture conditions. The two tracers differ in efficiency of tracing different sensory neurons subpopulations: while WGA is more efficient in tracing small C-fibres (CGRP-positive), CTB is more efficient in tracing A-fibres (NF200+) of sensory neurons. In summary, we have successfully established retrograde tracing techniques for sensory neurons towards studying and targeting breast cancer innervation.

8.
Bioorg Chem ; 147: 107395, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38705105

RESUMEN

Fluorination of carbohydrate ligands of lectins is a useful approach to examine their binding profile, improve their metabolic stability and lipophilicity, and convert them into 19F NMR-active probes. However, monofluorination of monovalent carbohydrate ligands often leads to a decreased or completely lost affinity. By chemical glycosylation, we synthesized the full series of methyl ß-glycosides of N,N'-diacetylchitobiose (GlcNAcß(1-4)GlcNAcß1-OMe) and LacdiNAc (GalNAcß(1-4)GlcNAcß1-OMe) systematically monofluorinated at all hydroxyl positions. A competitive enzyme-linked lectin assay revealed that the fluorination at the 6'-position of chitobioside resulted in an unprecedented increase in affinity to wheat germ agglutinin (WGA) by one order of magnitude. For the first time, we have characterized the binding profile of a previously underexplored WGA ligand LacdiNAc. Surprisingly, 4'-fluoro-LacdiNAc bound WGA even stronger than unmodified LacdiNAc. These observations were interpreted using molecular dynamic calculations along with STD and transferred NOESY NMR techniques, which gave evidence for the strengthening of CH/π interactions after deoxyfluorination of the side chain of the non-reducing GlcNAc. These results highlight the potential of fluorinated glycomimetics as high-affinity ligands of lectins and 19F NMR-active probes.


Asunto(s)
Disacáridos , Aglutininas del Germen de Trigo , Disacáridos/química , Disacáridos/síntesis química , Aglutininas del Germen de Trigo/química , Aglutininas del Germen de Trigo/metabolismo , Halogenación , Estructura Molecular , Acetilglucosamina/química , Acetilglucosamina/metabolismo , Lactosa/análogos & derivados
9.
Nutr Res ; 127: 13-26, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38820937

RESUMEN

Wheat germ (WG), a by-product of flour milling, is rich in bioactive substances that may help improve health complications associated with increased adiposity. This study investigated the effects of WG on gut health, metabolic, and inflammatory markers in adults classified as overweight. We hypothesized that WG, because of its many bioactive components, would improve gut health and metabolic, and inflammatory markers in overweight adults. Forty adults (18-45 years old) and with a body mass index between 25 and 30 kg/m2 participated in this single-blinded randomized controlled pilot study. Participants consumed the study supplements containing 30 g of either cornmeal (control, CL) or WG daily for 4 weeks. Primary outcome variables were gut health markers including gut microbiota, gut integrity markers, and fecal short-chain fatty acids, whereas secondary outcome variables included metabolic and inflammatory parameters assessed at baseline and at the end of supplementation. Thirty-nine participants (n = 19 and 20 for CL and WG group, respectively) completed the study. The genus Faecalibacterium was significantly higher in the WG group compared to CL post-supplementation but no significant changes in other gut health markers, short-chain fatty acids, inflammatory markers, and lipid profiles were observed. Compared with baseline, WG improved markers of glucose homeostasis including insulin (P = .02), homeostatic model assessment of insulin resistance (P = .03), glycated hemoglobin (P = .07), and the pro-inflammatory adipokine, resistin (P = .04). However, these parameters after intervention were not different with control. Our findings suggest that WG supplementation have modest effects on gut health but may provide an economical option for individuals to improve glycemic control.


Asunto(s)
Biomarcadores , Glucemia , Suplementos Dietéticos , Microbioma Gastrointestinal , Homeostasis , Sobrepeso , Triticum , Humanos , Adulto , Proyectos Piloto , Masculino , Femenino , Persona de Mediana Edad , Biomarcadores/sangre , Adulto Joven , Glucemia/metabolismo , Método Simple Ciego , Adolescente , Heces/microbiología , Heces/química , Ácidos Grasos Volátiles , Índice de Masa Corporal , Resistencia a la Insulina
10.
Photosynth Res ; 161(1-2): 127-140, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38662326

RESUMEN

It has been thoroughly documented, by using 31P-NMR spectroscopy, that plant thylakoid membranes (TMs), in addition to the bilayer (or lamellar, L) phase, contain at least two isotropic (I) lipid phases and an inverted hexagonal (HII) phase. However, our knowledge concerning the structural and functional roles of the non-bilayer phases is still rudimentary. The objective of the present study is to elucidate the origin of I phases which have been hypothesized to arise, in part, from the fusion of TMs (Garab et al. 2022 Progr Lipid Res 101,163). We take advantage of the selectivity of wheat germ lipase (WGL) in eliminating the I phases of TMs (Dlouhý et al. 2022 Cells 11: 2681), and the tendency of the so-called BBY particles, stacked photosystem II (PSII) enriched membrane pairs of 300-500 nm in diameter, to form large laterally fused sheets (Dunahay et al. 1984 BBA 764: 179). Our 31P-NMR spectroscopy data show that BBY membranes contain L and I phases. Similar to TMs, WGL selectively eliminated the I phases, which at the same time exerted no effect on the molecular organization and functional activity of PSII membranes. As revealed by sucrose-density centrifugation, magnetic linear dichroism spectroscopy and scanning electron microscopy, WGL disassembled the large laterally fused sheets. These data provide direct experimental evidence on the involvement of I phase(s) in the fusion of stacked PSII membrane pairs, and strongly suggest the role of non-bilayer lipids in the self-assembly of the TM system.


Asunto(s)
Complejo de Proteína del Fotosistema II , Tilacoides , Complejo de Proteína del Fotosistema II/metabolismo , Tilacoides/metabolismo , Espectroscopía de Resonancia Magnética , Lípidos/química , Fusión de Membrana/fisiología
11.
Appl Environ Microbiol ; 90(5): e0012224, 2024 May 21.
Artículo en Inglés | MEDLINE | ID: mdl-38567954

RESUMEN

Saprotrophic fungi that cause brown rot of woody biomass evolved a distinctive mechanism that relies on reactive oxygen species (ROS) to kick-start lignocellulosic polymers' deconstruction. These ROS agents are generated at incipient decay stages through a series of redox relays that shuttle electrons from fungus's central metabolism to extracellular Fenton chemistry. A list of genes has been suggested encoding the enzyme catalysts of the redox processes involved in ROS's function. However, navigating the functions of the encoded enzymes has been challenging due to the lack of a rapid method for protein synthesis. Here, we employed cell-free expression system to synthesize four redox or degradative enzymes, which were identified, by transcriptomic data, as conserved players of the ROS oxidation phase across brown rot fungal species. All four enzymes were successfully expressed and showed activities that enable confident assignment of function, namely, benzoquinone reductase (BQR), ferric reductase, α-L-arabinofuranosidase (ABF), and heme-thiolate peroxidase (HTP). Detailed analysis of their catalytic features within the context of brown rot environments allowed us to interpret their roles during ROS-driven wood decomposition. Specifically, we validated the functions of BQR as the driver redox enzyme of Fenton cycles and reconstructed its interactions with the co-occurring HTP or laccase and ABF. Taken together, this research demonstrated that the cell-free expression platform is adequate for synthesizing functional fungal enzymes and provided an alternative route for the rapid characterization of fungal proteins, escalating our understanding of the distinctive biocatalyst system for plant biomass conversion.IMPORTANCEBrown rot fungi are efficient wood decomposers in nature, and their unique degradative systems harbor untapped catalysts pursued by the biorefinery and bioremediation industries. While the use of "omics" platforms has recently uncovered the key "oxidative-hydrolytic" mechanisms that allow these fungi to attack lignocellulose, individual protein characterization is lagging behind due to the lack of a robust method for rapid synthesis of crucial fungal enzymes. This work delves into the studies of biochemical functions of brown rot enzymes using a rapid, cell-free expression platform, which allowed the successful depictions of enzymes' catalytic features, their interactions with Fenton chemistry, and their roles played during the incipient stage of brown rot when fungus sets off the reactive oxygen species for oxidative degradation. We expect this research could illuminate cell-free protein expression system's use to fulfill the increasing need for functional studies of fungal enzymes, advancing the discoveries of novel biomass-converting catalysts.


Asunto(s)
Biomasa , Proteínas Fúngicas , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Lignina/metabolismo , Sistema Libre de Células , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismo
12.
Artículo en Inglés | MEDLINE | ID: mdl-38656402

RESUMEN

The combination of shikonin (SKN) and gefitinib (GFB) can reverse the drug resistance of lung cancer cells by affecting energy metabolism. However, the poor solubility of SKN and GFB limits their clinical application because of low bioavailability. Wheat germ agglutinin (WGA) can selectively bind to sialic acid and N-acetylglucosamine on the surfaces of microfold cells and enterocytes, and is a targeted biocompatible material. Therefore, we created a co-delivery micelle system called SKN/GFB@WGA-micelles with the intestinal targeting functions to enhance the oral absorption of SKN and GFB by promoting mucus penetration for nanoparticles via oral administration. In this study, Caco-2/HT29-MTX-E12 co-cultured cells were used to simulate a mucus/enterocyte dual-barrier environment, and HCC827/GR cells were used as a model of drug-resistant lung cancer. We aimed to evaluate the oral bioavailability and anti-tumor effect of SKN and GFB using the SKN/GFB@WGA-micelles system. In vitro and in vivo experimental results showed that WGA promoted the mucus penetration ability of micelles, significantly enhanced the uptake efficiency of enterocytes, improved the oral bioavailability of SKN and GFB, and exhibited good anti-tumor effects by reversing drug resistance. The SKN/GFB@WGA-micelles were stable in the gastrointestinal tract and provided a novel safe and effective drug delivery strategy.

13.
Medicina (Kaunas) ; 60(4)2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38674285

RESUMEN

Background and Objectives: TAA is potent hepatic/renal toxicant. Conversely, WGO is a potent dietary supplement with impressive antioxidant properties. Olmutinib is an apoptotic chemotherapy drug that does not harm the liver or kidney. This study investigated the impact of olmutinib and wheat germ oil (WGO) on Thioacetamide (TAA)-induced gene alterations in mice liver and kidney tissues. Materials and Methods: Adult male C57BL/6 mice were exposed to 0.3% TAA in drinking water for 14 days, followed by the oral administration of olmutinib (30 mg/kg) and WGO (1400 mg/kg) for 5 consecutive days. Treatment groups included the following: groups I (control), II (TAA-exposed), III (TAA + olmutinib), IV (TAA + WGO), and V (TAA + olmutinib + WGO). Results: The findings revealed that TAA exposure increased MKi67 and CDKN3 gene expression in liver and kidney tissues. Olmutinib treatment effectively reversed these TAA-induced effects, significantly restoring MKi67 and CDKN3 gene expression. WGO also reversed MKi67 effects in the liver but exhibited limited efficacy in reversing CDKN3 gene alterations induced by TAA exposures in both the liver and kidney. TAA exposure showed the tissue-specific expression of TP53, with decreased expression in the liver and increased expression in the kidney. Olmutinib effectively reversed these tissue-specific alterations in TP53 expression. While WGO treatment alone could not reverse the gene alterations induced by TAA exposure, the co-administration of olmutinib and WGO exhibited a remarkable potentiation of therapeutic effects in both the liver and kidney. The gene interaction analysis revealed 77.4% of physical interactions and co-localization between MKi67, CDKN3, and TP53 expressions. Protein-protein interaction networks also demonstrated physical interactions between MKi67, TP53, and CDKN3, forming complexes or signaling cascades. Conclusions: It was predicted that the increased expression of the MKi67 gene by TAA leads to the increase in TP53, which negatively regulates the cell cycle via increased CDKN3 expression in kidneys and the restoration of TP53 levels in the liver. These findings contribute to our understanding of the effects of olmutinib and WGO on TAA-induced gene expression changes and highlight their contrasting effects based on cell cycle alterations.


Asunto(s)
Riñón , Hígado , Ratones Endogámicos C57BL , Tioacetamida , Animales , Ratones , Riñón/efectos de los fármacos , Riñón/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Ciclo Celular/efectos de los fármacos , Triticum
14.
Chem Pharm Bull (Tokyo) ; 72(3): 258-265, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38432907

RESUMEN

Glycated albumin (GA) is one of the proteins that replaces several sugar moieties and can be used as an indicator of diabetes mellitus. We developed a sensing system that uses GA in the early detection of diabetes mellitus. In this study, H6Y4C acetylated (Ac-) at the N-terminals of the peptide was combined with wheat germ agglutinin (WGA) to recognize glucose moieties. The Ac-H6Y4C-WGA was constructed as a GA-sensing probe. The tyrosine residues of Y4C exhibited an oxidation peak, and His-tag moieties were introduced to separate Ac-H6Y4C-WGA in the synthesis of the probe. The Ac-H6Y4C-WGA probe binds with the 1-2 molecules of Ac-H6Y4C per WGA using matrix assisted laser desorption/ionization-time of flight (MALDI-TOF)-MS. Next, the functions of Ac-H6Y4C-WGA were evaluated using voltammetry. The number of electron-transfers was calculated based on the relationship between the peak potential and logarithm of scan rate and was 3.03. In the electrochemical measurements with mannose and bovine serum albumin, the peak currents were similar to that of GA alone. By contrast, a decrease in the peak current was suppressed when glucose was added to the solution containing the probe. As a result, Ac-H6Y4C-WGA was selectively bound to the glucose moieties of GA. The calibration curve via differential pulse voltammetry was proportional to the concentrations of GA and ranged from 1.0 × 10-12 to 2.0 × 10-11 M with a detection limit of 3.3 × 10-13 M.


Asunto(s)
Diabetes Mellitus , Albúmina Sérica , Humanos , Diabetes Mellitus/diagnóstico , Electrones , Glucosa , Péptidos , Albúmina Sérica/química , Técnicas Biosensibles/métodos
15.
Mikrochim Acta ; 191(4): 209, 2024 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-38499840

RESUMEN

A novel colorimetric platform was designed for the determination of S. aureus by utilizing a dual-recognition strategy, where wheat germ agglutinin (WGA)-functionalized magnetic beads were served as separation elements to capture and enrich S. aureus efficiently from the matrix. Horseradish peroxidase (HRP) labeled chicken anti-protein A IgY (HRP-IgY) was used to label the captured S. aureus. A chicken IgY was introduced as a signal tracer to bind with staphylococcal protein A (SPA) on the surface of S. aureus, which can circumvent the interference from protein G-producing Streptococcus. Subsequently, the colorimetric signal was achieved by an HRP-catalyzed reaction, which was amplified by HRP-IgY bound by approximately 80,000 SPA molecules on one S. aureus. The entire detection process could be accomplished within 90 min. Under optimal conditions, the linear response of different S. aureus concentrations ranged from 7.8 × 102 to 2.0 × 105 CFU/mL and the limit of detection reached down to 3.9 × 102 CFU/mL. Some common non-target bacteria yielded negative results, indicating the excellent specificity of the method. The developed strategy was successfully applied to the determination of S. aureus in various types of samples with satisfactory recoveries. Therefore, the novel dual-recognition strategy possessed the advantages of high sensitivity, specificity, and low cost and exhibited considerable potential as a promising tool to defend public health.


Asunto(s)
Infecciones Estafilocócicas , Staphylococcus aureus , Humanos , Aglutininas del Germen de Trigo , Colorimetría/métodos , Inmunoglobulinas , Infecciones Estafilocócicas/diagnóstico , Infecciones Estafilocócicas/microbiología , Peroxidasa de Rábano Silvestre/metabolismo
16.
Cureus ; 16(1): e52432, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38371095

RESUMEN

BACKGROUND AND OBJECTIVE: Nigella sativa (NS) oil has been used as an ointment for relief from abscesses, nasal ulcers, orchitis, eczema, and swollen joints. The nutritional and biological values of wheat germ oil (WGO) are imperative points for testing its wound healing properties in traumatic ulcer. The aim of the study was to evaluate and compare the ability of NS versus WGO in promoting the healing of induced traumatic ulcer in albino rats clinically and histologically. MATERIALS AND METHODS: This study was carried out after the approval of the Research Ethics Committee (REC) of the Faculty of Dentistry, Suez Canal University, in Ismailia, Egypt, on 60 albino rats with induced labial ulcer according to calculated sample size. All animals were anaesthetized with an intraperitoneal injection of 10% ketamine. The ulcer was produced on the labial mucosa corresponding to the midline between the lower two incisors of each rat. After induction of the ulcer, rats were randomly divided into four groups according to the treatment medicament: Group A (negative control group): 15 rats which remained without treatment; Group B (positive control): 15 rats which received daily a topical application of 1 ml of cetylpyridinium chloride (CPC) and lidocaine gel; Group C (NS group): 15 rats which received a daily topical application of 1 mm of NS oil painted by a brush covering the whole area of the ulcer; and Group D (WGO group): 15 rats which received 1 mm of WGO. The ulcers were measured using a digital caliper and were recorded using a digital camera at days 0, 3, 7, and 9, the largest (D) and smallest (d) diameters of the lesion were recorded, and the ulcer area was calculated using the following formula: A=π×D/2×d/2. Tissue samples were taken for histological examination, and the labial mucosa was dissected out and embedded in paraffin wax blocks. The blocks were cut with microtome to obtain sections of 4-5 µm thickness to be stained with hematoxylin and eosin stain and Masson's trichrome stain. All sections were examined under a light microscope, and the presence of inflammatory cells and collagen tissue remodeling were evaluated. RESULTS: Within the control group, there are statistically non-significant changes in the mean of the surface area of ulcer when comparing changes in 10 rats who survived till the seventh day and inflammatory cell count when comparing changes in five rats who were sacrificed at the seventhday. There was a significant decrease in surface area and inflammatory cell count in five rats who survived till the ninth day. Within the WGO group only, all survived rats had healed ulcer at the ninth day. There is a significant decrease in inflammatory cell count in five rats who survived till the ninth day. CONCLUSION: WGO was significantly more effective in the treatment of animal-induced ulcer compared to NS oil or CPC and lidocaine oral gel.

17.
J Anim Physiol Anim Nutr (Berl) ; 108(3): 806-815, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38311826

RESUMEN

This experiment aimed to evaluate the replacement of cottonseed meal (CSM) with wheat germ meal (WGM) in diets of growing lambs on feed utilization and growth performance. Twenty-eight Ossimi male lambs (38 ± 0.8 kg weight), and 180 ± 5 days were divided randomly into four experimental groups in a complete randomized design for 105 days. Cottonseed meal was replaced with WGM at 0 (WGM0 treatment), 50 (WGM50 treatment), 75 (WGM75 treatment) and 100% (WGM100 treatment). The chemical analysis of the total essential and non-essential amino acids showed an increase at the WGM diet compared to CSM. The replacement of CSM with WGM linearly and quadratically improved (p ˂ 0.05) lambs' growth performance and feed conversion. The WGM50 and WGM100 treatments lowered (p ˂ 0.05) feed intake, without affecting nutrient digestibility or diets' nutritive. Feeding WGM increased (p ˂ 0.05) total protein, albumin, and urea-N concentrations in blood of lambs. The WGM100 treatment showed the highest relative percentage of net revenue compared to the other treatments. It is concluded that the complete replacement of CSM with WGM showed positive effects on lambs' performance and economic efficiency.


Asunto(s)
Alimentación Animal , Fenómenos Fisiológicos Nutricionales de los Animales , Aceite de Semillas de Algodón , Dieta , Triticum , Animales , Masculino , Alimentación Animal/análisis , Aceite de Semillas de Algodón/administración & dosificación , Aceite de Semillas de Algodón/química , Dieta/veterinaria , Ovinos/fisiología , Ovinos/crecimiento & desarrollo , Triticum/química
18.
Drug Deliv Transl Res ; 14(3): 621-636, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-37787882

RESUMEN

This study aimed to enhance the oral absorption of Panax notoginseng saponins (PNS) via nanoparticles modified with thiolated trimethyl chitosan (TMC-Cys) and wheat germ agglutinin (WGA), termed PP-WT NPs. In vitro investigations revealed that PP-WT NPs exhibited delayed release of PNS and a strong tolerance to the gastric acids and digestive enzymes. Moreover, PP-WT NPs exhibited efficient cellular uptake and transport capabilities in the Caco-2/HT29-co-cultured cell model. In vivo animal experiments demonstrated that PP-WT NPs effectively overcame the mucus layer barrier, with the effective permeability coefficients of R1, Rg1, and Rb1 in the small intestine being 1.68, 1.64, and 1.63 times higher than those of free PNS, respectively. Taken together, thiolated trimethyl chitosan and wheat germ agglutinin-modified nanoparticles hold significant potential for improving the oral absorption of PNS, representing an attractive strategy for enhanced therapeutic efficacy.


Asunto(s)
Quitosano , Panax notoginseng , Saponinas , Ratas , Humanos , Animales , Ratas Sprague-Dawley , Células CACO-2
19.
Food Sci Nutr ; 11(11): 7242-7254, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37970387

RESUMEN

Polyamines have received a lot of attention since the 1990s because of their anti-aging, anti-chronic disease, and proliferative effects. Wheat germ was reported as one of the natural sources of high polyamine, especially spermidine. The current study used three types of wheat germ: group A was industrially separated germ from whole grain, group B was the commercially available germinated wheat germ, and group C was manually separated wheat germ from germinated grain. The polyamine content of putrescine, spermidine, and spermine has been determined using a simplified isocratic LC-MS/MS method. An optimized extraction procedure was performed on all seven samples for obtaining a polyamine-enriched extract. The three dominant carbomylated polyamines were identified by analyzing the extracted samples in order to determine their relative abundance. Wheat germ powders contain the highest amount of polyamines (220-337 µg/g) of which spermidine is one of the most important. Germinated wheat grains, on the other hand, contain the least amount of this polyamine. The commercially available separated wheat germs are suggested as a good nutrition source of these polyamines.

20.
Nutr Res ; 120: 135-144, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38000279

RESUMEN

Evidence has demonstrated that oxidative stress plays a crucial role in regulating cellular glucose metabolism. In previous studies, wheat germ peptide (WGP) was found to effectively mitigate oxidative stress induced by high glucose. Based on the information provided, we hypothesized that WGP could exhibit antihyperglycemic and anti-insulin-resistant effects in cells. The insulin-resistant cell model was established by insulin stimulation. The glucose consumption, glycogen content, and the activities of hexokinase and pyruvate kinase following WGP treatment were measured. The protein expression of SOCS3, phosphorylated insulin receptor substrate-1 (p-IRS1), IRS1, phosphorylated protein kinase B (p-Akt), Akt, glucose transporter 2 (GLUT2), phosphorylated GSK 3ß, GSK 3ß, FOXO1, G6P, and phosphoenolpyruvate carboxykinase were assessed by western blot analysis. Our results demonstrated that WGP treatment increased cellular glucose consumption and glycogen synthesis and enhanced hexokinase and pyruvate kinase activities. Additionally, WGP treatment was observed to cause a significant reduction in the expression of SOCS3, FOXO1, G6P, and phosphoenolpyruvate carboxykinase, as well as in the ratio of p-IRS1/IRS1. Conversely, the expression of GLUT2 and the ratios of p-Akt/Akt and p-GSK3ß/GSK3ß were upregulated by WGP. These findings suggested that WGP can activate the SOCS3/IRS1/Akt signaling pathway, thus promoting the phosphorylation of GSK-3ß and increasing the expression of FOXO1 and GLUT2, which contribute to enhancing glycogen synthesis, inhibiting gluconeogenesis, and promoting glucose transport in insulin-resistant HepG2 cells.


Asunto(s)
Resistencia a la Insulina , Proteínas Proto-Oncogénicas c-akt , Humanos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Glucógeno Sintasa Quinasa 3 beta/farmacología , Triticum , Proteínas Sustrato del Receptor de Insulina/metabolismo , Hexoquinasa/metabolismo , Hexoquinasa/farmacología , Piruvato Quinasa/metabolismo , Fosfoenolpiruvato/metabolismo , Fosfoenolpiruvato/farmacología , Hepatocitos/metabolismo , Glucosa/metabolismo , Insulina/metabolismo , Glucógeno/metabolismo , Proteína 3 Supresora de la Señalización de Citocinas/metabolismo
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