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Curcumin, a bioactive compound, showed versatile in anti-inflammatory and anti-cancer ability, while their biological fate in elderly is unclear. In this study, curcumin-loaded nanoparticles based on octyl succinate hydrate (OSA) starch and sodium caseinate were prepared and the in vitro elderly digestion and absorption fate was investigated. The loading capacity of curcumin-loaded nanoparticles prepared from OSA starch (HI), sodium caseinate (SC) and OSA starchsodium caseinate (HS) were all higher than 15%. Curcumin release behavior of the three nanoparticles during in vitro digestion conformed to first-order kinetics. Meanwhile, the transport efficiency of curcumin for HI, SC, and HS increased significantly than the free curcumin (near 1-fold), and the permeability were 1.9, 2.0, and 2.0 times, respectively. The gene expressions of TNF-α, SREBP2 and NPC1L1 in the organoids were enhanced than control group. This study provided scientific reference and guidance for encapsulation of curcumin and digestion and absorption properties in elderly.
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Curcumina , Digestão , Nanopartículas , Curcumina/química , Curcumina/metabolismo , Curcumina/farmacologia , Nanopartículas/química , Humanos , Organoides/metabolismo , Portadores de Fármacos/química , Amido/química , Amido/metabolismoRESUMO
In this study, carrageenan (CG), xanthan gum (XG) and locust bean gum (LBG), which can be used in infant formulas in China national standards, were selected to prepare LF-polysaccharide complexes to improve the stability of lactoferrin. The results showed that LF interacted more strongly with polysaccharides and did not affect the LF structure to a large extent when the pH and protein/polysaccharide mass ratio were 7 and 10:1 for LF-CG, 8 and 5:1 for LF-XG, 7 and 15:1 for LF-LBG. The zeta potential and fluorescence intensity of the LF-polysaccharide complexes displayed a decreasing trend with the increase in pH. When pH < 6, LF-CG and LF-XG exhibited precipitation and increased UV absorbance. Complexation between LF and CG/XG mainly attributed to electrostatic interactions, while LF and LBG form complexes based on hydrogen bonding or hydrophobic interactions. This study could provide a reference for the practical application of LF in infant formula.
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Fórmulas Infantis , Lactoferrina , Polissacarídeos , Lactoferrina/química , Concentração de Íons de Hidrogênio , Polissacarídeos/química , Fórmulas Infantis/química , Galactanos/química , Polissacarídeos Bacterianos/química , Gomas Vegetais/química , Mananas/química , Humanos , Carragenina/químicaRESUMO
Thymic stromal lymphopoietin (TSLP) is a member of the IL-2 cytokine family and has been widely recognized as a master regulator of type 2 inflammatory responses at barrier surfaces. Recent studies found dysregulation of the TSLP-TSLP receptor (TSLPR) pathway is associated with the pathogenesis of not only allergic diseases but also a wide variety of cancers including both solid tumors and hematological tumors. Thus, the blockade of TSLP represents an attractive therapeutic strategy for allergic diseases and cancer. In this study, we report the development of a novel humanized anti-TSLP monoclonal antibody (mAb) HZ-1127. Binding affinity, specificity, and ability of HZ-1127 in inhibiting TSLP were tested. HZ-1127 selectively binds to the TSLP cytokine with high affinity and specificity. Furthermore, HZ-1127 dramatically inhibits TSLP-dependent STAT5 activation and is more potent than Tezepelumab, which is an FDA-approved humanized mAb against TSLP for severe asthma treatment in inhibiting TSLP-induced CCL17 and CCL22 chemokines secretion in human peripheral blood mononuclear cells. Our pre-clinical study demonstrates that HZ-1127 may serve as a potential therapeutic agent for allergic diseases and cancer.
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Controlling the structure and viscosity of food can influence the development of diet-related diseases. Food viscosity has been linked with health through its impact on human digestion and gastrointestinal transit, however, there is limited understanding of how the viscosity of food regulates gastric emptying. Here, we used model food preparations with different viscosities using guar gum, to explore the mechanism underlying the influence of viscosity on gastric motility, gastric emptying and postprandial blood glucose. Based on experiments in human volunteers and animals, we demonstrated that high viscosity meals increased gastric antrum area and gastric retention rate. Viscosity also affected gut hormone secretion, reduced the gene expression level of interstitial cells of Cajal, resulting in a delay of gastric emptying and limiting the increase in postprandial glucose. This improved mechanistic understanding of food viscosity during gastric digestion is important for designing new foods to benefit human health.
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Galactanos , Esvaziamento Gástrico , Mananas , Gomas Vegetais , Humanos , Viscosidade , Mananas/química , Mananas/farmacologia , Gomas Vegetais/química , Galactanos/química , Galactanos/farmacologia , Animais , Masculino , Período Pós-Prandial , Adulto , Glicemia/metabolismo , Feminino , Alimentos , Camundongos , DigestãoRESUMO
BACKGROUND: Hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs) play a pivotal role in maintaining lifelong hematopoiesis. The distinction between stem cells and other progenitors, as well as the assessment of their functions, has long been a central focus in stem cell research. In recent years, deep learning has emerged as a powerful tool for cell image analysis and classification/prediction. METHODS: In this study, we explored the feasibility of employing deep learning techniques to differentiate murine HSCs and MPPs based solely on their morphology, as observed through light microscopy (DIC) images. RESULTS: After rigorous training and validation using extensive image datasets, we successfully developed a three-class classifier, referred to as the LSM model, capable of reliably distinguishing long-term HSCs, short-term HSCs, and MPPs. The LSM model extracts intrinsic morphological features unique to different cell types, irrespective of the methods used for cell identification and isolation, such as surface markers or intracellular GFP markers. Furthermore, employing the same deep learning framework, we created a two-class classifier that effectively discriminates between aged HSCs and young HSCs. This discovery is particularly significant as both cell types share identical surface markers yet serve distinct functions. This classifier holds the potential to offer a novel, rapid, and efficient means of assessing the functional states of HSCs, thus obviating the need for time-consuming transplantation experiments. CONCLUSION: Our study represents the pioneering use of deep learning to differentiate HSCs and MPPs under steady-state conditions. This novel and robust deep learning-based platform will provide a basis for the future development of a new generation stem cell identification and separation system. It may also provide new insight into the molecular mechanisms underlying stem cell self-renewal.
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Aprendizado Profundo , Animais , Camundongos , Células-Tronco Hematopoéticas/metabolismo , Hematopoese , Células-Tronco Multipotentes , Diferenciação CelularRESUMO
Lactoferrin (LF) is a thermally sensitive iron-binding globular glycoprotein. Heat treatment can induce its denaturation and aggregation and thus affect its functional activity. In this study, carrageenan (CG), xanthan gum (XG) and locust bean gum (LBG), allowed to apply in infant food, were used to form protein-polysaccharide complexes to improve the thermal stability of LF. Meanwhile, in vitro simulated infant digestion and absorption properties of LF were also estimated. The results showed that the complexes formed by CG and XG with LF (LF-CG and LF-XG) could significantly inhibit the loss of α-helix structure of LF against heating. LF-CG and LF-LBG could protect LF from digestion in simulated infant gastric fluid and slow down the degradation of LF under the simulated intestinal conditions. Besides, LF, LF-CG and LF-XG showed no adverse effects on the growth of Caco-2 cells in the LF concentration range of 10-300 µg/mL, and LF-XG exhibited better beneficial to improve the cell uptake of the digestive product than the other protein-polysaccharides at the LF concentration of 100 µg/mL. This study may provide a reference for the enhancement of thermal processing stability of LF and development infant food ingredient with high nutrients absorption efficiency in the gastrointestinal environment in the future.
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Trato Gastrointestinal , Lactoferrina , Lactente , Humanos , Lactoferrina/química , Células CACO-2 , Fenômenos Químicos , Trato Gastrointestinal/metabolismoRESUMO
Inspired by membrane structure of breast milk and infant formula fat globules, four liposomes with different particle size (large and small) and compositions (Single phospholipids contained phosphatidylcholine, complex phospholipids contained phosphatidylcholine, phosphatidylethanolamine and sphingomyelin) were fabricated to deliver lactoferrin and DHA. In vitro infant semi-dynamic digestive behavior and absorption in intestinal organoids of liposomes were investigated. Liposomal structures were negligible changed during semi-dynamic gastric digestion while damaged in intestine. Liposomal degradation rate was primarily influenced by particle size, and complex phospholipids accelerated DHA hydrolysis. The release rate of DHA (91.7 ± 1.3 %) in small-sized liposomes (0.181 ± 0.001 µm) was higher than free DHA (unencapsulated, 64.6 ± 3.4 %). Complex phospholipids liposomal digesta exhibited higher transport efficiency (3.4-fold for fatty acids and 2.0-fold for amino acids) and better organoid growth than digesta of bare nutrients. This study provided new insights into membrane structure-functionality relationship of liposomes and may aid in the development of novel infant nutrient carriers.
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Lactoferrina , Lipossomos , Lactente , Feminino , Humanos , Animais , Suínos , Lipossomos/química , Lactoferrina/química , Fosfolipídeos/química , Fosfatidilcolinas , Digestão , Ácidos Docosa-HexaenoicosRESUMO
A smart film was developed to detect the freshness of pork by incorporating blueberry anthocyanins (BAs) and hinokitiol (HIN) loaded zeolite-imidazolium framework (HIN@ZIF-8) with into a sodium alginate matrix, and its microstructure and physicochemical properties were studied. The SA matrix was doped with BAs and HIN@ZIF-8 nanoparticles (SA-BAs/HIN@ZIF-8) to increase its tensile strength and reduce its water vapor permeability. HIN@ZIF-8 has low cytotoxicity, and SA-BAs/HIN@ZIF-8 membranes have long-lasting antimicrobial and highly sensitive color development properties against Escherichia coli and Staphylococcus aureus. The results of pork preservation experiments showed that SA-BA/HIN@ZIF-8 could extend the shelf life of pork to 6 days at 4 â. E-nose evaluation experiments showed that SA-BAs/HIN@ZIF-8 could inhibit compounds that cause unpleasant and irritating odours. Therefore, SA-BAs/HIN@ZIF-8 was considered to be an effective method to improve the freshness of pork, and the results showed that it has a promising application in food preservation.
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Mirtilos Azuis (Planta) , Monoterpenos , Nanopartículas , Carne de Porco , Carne Vermelha , Tropolona/análogos & derivados , Suínos , Animais , Alginatos/farmacologia , Antocianinas/farmacologia , Antibacterianos/farmacologia , Escherichia coli , Embalagem de Alimentos , Concentração de Íons de HidrogênioRESUMO
Gallic acid (GA) and epigallocatechin gallate (EGCG), cooperated at varied ratios (1:0, 3:1, 1:1, 1:3, and 0:1), were employed to modify gel properties of calcium induced-whey protein emulsion gel. The effects of GA/EGCG on emulsion morphology, as well as gel properties and in vitro digestive behavior of the emulsion gels were investigated. Compared with emulsions without phenolics, GA/EGCG induced slightly smaller particle size and stronger electrostatic repulsion between emulsion droplets. Moreover, GA/EGCG, notably at a ratio of 3:1, promoted electrostatic and hydrophobic interactions between protein molecules and the formation of a compact and filamentous gel microstructure, resulting in a remarkable increment in the gel strength (up to 106 %). Furthermore, in vitro oral digestion, dynamic gastric digestion (using an artificial gastric digestive system, AGDS), and intestinal digestion of the emulsion gels were simulated. Particle size and protein hydrolysis results revealed that GA/EGCG was prone to weaken the physical disintegration of gels, reduce protein hydrolysis, and enhance the stability of emulsified oil droplets during dynamic gastric digestion. As a consequence, delayed release of oil droplets was observed in the gels and more free fatty acids were released in the intestinal digestion, particularly in the gel with GA/EGCG (3:1). These findings would provide novel strategies for application of phenolic compounds in developing protein gel-based delivery systems.
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Digestão , Ácido Gálico , Proteínas do Soro do Leite/química , Emulsões/química , Géis/químicaRESUMO
Background: Hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs) play a pivotal role in maintaining lifelong hematopoiesis. The distinction between stem cells and other progenitors, as well as the assessment of their functions, has long been a central focus in stem cell research. In recent years, deep learning has emerged as a powerful tool for cell image analysis and classification/prediction. Methods: In this study, we explored the feasibility of employing deep learning techniques to differentiate murine HSCs and MPPs based solely on their morphology, as observed through light microscopy (DIC) images. Results: After rigorous training and validation using extensive image datasets, we successfully developed a three-class classifier, referred to as the LSM model, capable of reliably distinguishing long-term HSCs (LT-HSCs), short-term HSCs (ST-HSCs), and MPPs. The LSM model extracts intrinsic morphological features unique to different cell types, irrespective of the methods used for cell identification and isolation, such as surface markers or intracellular GFP markers. Furthermore, employing the same deep learning framework, we created a two-class classifier that effectively discriminates between aged HSCs and young HSCs. This discovery is particularly significant as both cell types share identical surface markers yet serve distinct functions. This classifier holds the potential to offer a novel, rapid, and efficient means of assessing the functional states of HSCs, thus obviating the need for time-consuming transplantation experiments. Conclusion: Our study represents the pioneering use of deep learning to differentiate HSCs and MPPs under steady-state conditions. With ongoing advancements in model algorithms and their integration into various imaging systems, deep learning stands poised to become an invaluable tool, significantly impacting stem cell research.
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Salmonella can cause severe foodborne diseases. This study investigated the prevalence of Salmonella spp. in fresh foods in Hangzhou market and their harborage of antibiotic resistance and virulence genes, antibiotic susceptibility, and pathogenicity. A total of 500 samples (pork, n = 140; chicken, n = 128; vegetable, n = 232) were collected over a one-year period. Salmonella was found in 4.2% (21) of samples with the detection rate in pork, chicken and vegetables as 4.3% (6), 6.3% (8), and 3% (7), respectively. One Salmonella strain was recovered from each positive sample. The isolates were identified as six serotypes, of which S. Enteritidis (n = 7) and S. Typhimurium (n = 6) were the most predominant serotypes. The majority of isolates showed resistance to tetracycline (85.7%) and/or ciprofloxacin (71.4%). Tetracycline resistance genes showed the highest prevalence (90.5%). The occurrence of resistance genes for ß-lactams (blaTEM-1, 66.7%; and blaSHV, 9.5%) and aminoglycosides (aadA1, 47.6%; Aac(3)-Ia, 19%) was higher than sulfonamides (sul1, 42.9%) and quinolones (parC, 38.1%). The virulence gene fimA was detected in 57.1% of isolates. Gene co-occurrence analysis implied that resistance genes were associated with virulence genes. Furthermore, selected S. Typhimurium isolates (n = 4) carrying different resistance and virulence genes up-regulated the secretions of cytokines IL-6 and IL-8 by Caco-2 cells in different degrees, suggesting that virulence genes may play a role in inflammatory transcription. In in vivo virulence test, microbiological counts in mouse feces and tissues showed that all included S. Typhimurium were able to infect mice, with one strain showing significantly higher virulence than others. In conclusion, this study indicates Salmonella contamination in fresh foods in Hangzhou market poses a risk to public health and it should be closely monitored to prevent and control foodborne diseases.
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Antibacterianos , Doenças Transmitidas por Alimentos , Humanos , Animais , Camundongos , Antibacterianos/farmacologia , Virulência/genética , Células CACO-2 , Farmacorresistência Bacteriana/genética , Salmonella , Galinhas/microbiologia , Doenças Transmitidas por Alimentos/microbiologia , China/epidemiologia , Testes de Sensibilidade Microbiana , Farmacorresistência Bacteriana Múltipla/genéticaRESUMO
In this study, the effects of adding sodium tripolyphosphate during the extrusion of textured wheat protein (TWP)-based meat analogs were investigated. Five TWPs (TWP-C0, TWP-C0.10, TWP-C0.25, TWP-C0.50, and TWP-C0.75) were prepared with sodium tripolyphosphate concentrations of 0%, 0.10%, 0.25%, 0.50%, and 0.75%, respectively. The fibrous structure of TWPs was analyzed by determining their textural properties, degree of texturization, microstructure, and protein bonds. When the concentration of sodium tripolyphosphate increased from 0% to 0.75%, the fibers in TWPs became more regular and finer with smaller pores, the degree of texturization increased from 2.10 ± 0.09 to 2.73 ± 0.07, and the proportions of solubilized protein from the breaking of hydrophobic bonds and disulfide bonds increased from 2.06 ± 0.14% and 1.38 ± 0.11% to 3.42 ± 0.12% and 1.74 ± 0.05%, respectively. The results of particle size, soluble nitrogen content, and free amino acids of samples during digestion indicated that the disintegration rate and protein digestibility of TWPs increased with the increase in the concentration of sodium tripolyphosphate. After gastrointestinal digestion, the total free amino acids released in TWP-C0, TWP-C0.10, TWP-C0.25, TWP-C0.50, and TWP-C0.75 were 391.5 ± 2.2, 403.9 ± 1.5, 430.0 ± 3.6, 473.8 ± 2.9 and 485.3 ± 5.73 mg/10 g digesta, respectively. Sodium tripolyphosphate may improve the protein digestibility of TWPs by forming a finer fibrous structure with a more unfolded protein structure and more hydrophobic groups being exposed to enzymes.
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Digestão , Triticum , Triticum/química , Carne , Aminoácidos/metabolismoRESUMO
Chewing ability has a strong effect on food digestion. However, little is known about the relationship between the food mastication degree and the subsequent gastric emptying. This study was to explore the effects of individual chewing ability (strong and weak) on the in vivo oral processing characteristics and in vitro dynamic gastrointestinal starch hydrolysis of three types of rice (japonica rice, indica rice and waxy rice). Results showed that the swallowable bolus in the weak chewing group had larger holes and a looser microstructure with more small rice particles, while the strong chewing ones obtained a bolus with higher saliva content (up to 28%) and starch hydrolysis degree (up to 13.55%). Moreover, the gastric retention and starch hydrolysis of the strong chewing ability group were higher in the artificial gastric dynamic system (AGDS). The indica rice particles with the higher degree of fragmentation contacted enzymes easier and hydrolyzed quicker, thus emptying through the stomach faster (81.76%). However, the oral chewing properties of rice mainly influenced the starch digestion in the stomach and the initial stage of the small intestine (â¼5 min). This study suggested that the chewing ability and rice variety can influence the bolus properties, which in turn affected the gastric emptying and the degree of starch hydrolysis during digestion.
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Oryza , Amido , Amido/química , Hidrólise , Mastigação , Oryza/química , DigestãoRESUMO
In recent years, the blaNDM gene, which mediate resistance to carbapenems, has disseminated all over the world, and has also been detected in animals. Understanding the dissemination and accumulation of antibiotic resistance genes (ARGs) in a human-impacted environment is essential to solve the food safety problems caused by antibiotics. In this study, two strains of carbapenem bacteria carrying blaNDM were screened from 244 strains isolated from two T. sinensis farms in Zhejiang province, China. After their plasmids were isolated and sequenced, their structure and gene environment were analyzed and the mechanism of blaNDM gene transfer was explored. The study measured the fitness cost of plasmids carrying different blaNDM subtypes by four biological characteristics experiments. The results showed that the fitness cost of IncC plasmid carrying blaNDM-1 was higher than that of IncX3 plasmid carrying blaNDM-5. Furthermore, the real-time PCR showed that the decrease of transcription level of fitness-related genes lead to the different fitness cost of plasmids carrying different blaNDM subtypes. Fitness of many blaNDM-harboring plasmids enhanced the further dissemination of this gene and increase the risk of blaNDM gene spreading in aquatic environment, and thus further investigation of carbapenem-resistant bacterias among food animals are in urgent need.
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Escherichia coli , beta-Lactamases , Animais , Humanos , beta-Lactamases/genética , Plasmídeos/genética , Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Fazendas , Aquicultura , Testes de Sensibilidade MicrobianaRESUMO
Liposomes have become a research hotspot in recent years as food delivery systems with attractive properties, including the bilayer structure assembled like the cell membrane, reducing the side-effect and improving environmental stability of cargos, controlling release, extending duration of functional ingredients, and high biodegradable and biocompatible abilities in the body. However, the conventional liposomes lack stability during storage and are weak in targeted absorption in the gastrointestinal track. At present, surface modification has been approved to be an effective platform to shield these barricades and help liposomes deliver the agents safely and effectively to the ideal site. In this review, the gastrointestinal stability of conventional liposomes, cargo release models from liposomes, and the biological fate of the core materials after release were emphasized. Then, the strategies in both physical and chemical perspectives to improve the stability and utilization of liposomes in the gastrointestinal tract, and the emerging approaches for improving gut targeting by specifically modified liposomes and the intestinal receptors relative to liposomes/cargos absorption were highlighted. Last but not the least, the safety, challenges, and opportunities for the improvement of liposomal bioavailability were also discussed to inspire new applications of liposomes as oral carriers.
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Alimentos , Lipossomos , Lipossomos/química , Disponibilidade BiológicaRESUMO
As powerful bioactive compounds found in a variety of plant-based foods, (epi)catechins have been identified to be associated with an abundant array of health benefits. While their adverse impacts have also been gaining increasing attention, their intestinal impact is still unclear. In this study, intestinal organoids were used as an in vitro model to analyze the effects of four (epi)catechins on the development of the intestinal epithelial structure. Morphological characteristics, oxidative stress, and endoplasmic reticulum (ER) stress assays with (epi)catechins treatment showed that (epi)catechins promoted intestinal epithelial apoptosis and stress response. These effects had dose-dependent and structural differences (EGCG > EGC > ECG > EC). Furthermore, GSK2606414, a protein kinase RNA (PKR)-like ER kinase (PERK) pathway inhibitor, confirmed that the PERK-eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4)-C/EBP-homologous protein (CHOP) pathway is closely related to the damage. In addition, the results for the intestinal inflammatory mouse model further verified that (epi)catechins significantly delayed intestinal repair. Taken together, these findings revealed that overdosage of (epi)catechins has damage potential on the intestinal epithelium and may increase the risk of intestinal damage.
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Catequina , Estresse do Retículo Endoplasmático , Mucosa Intestinal , Estresse Oxidativo , Catequina/farmacologia , Animais , Camundongos , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/fisiopatologia , Fator de Iniciação 2 em Eucariotos , Organoides/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Transdução de Sinais , Enterite/fisiopatologiaRESUMO
Dysregulated elevation of interleukin-6 (IL-6) signaling is implicated in the pathogenesis of multiple pathophysiological states, and the functional neutralization of the IL-6 pathway with monoclonal antibodies has been proven an effective therapeutic method in treating various diseases with abnormally enhanced IL-6 signaling, and its clinical indications are expanding. Here, we report that by using the conventional hybridoma technology and humanization mutation method, we develop a novel humanized anti-IL-6 receptor (IL-6R) antibody-namely, HZ0412a. In our study, we found that HZ0412a exhibits higher binding affinity to soluble recombinant human IL-6R than tocilizumab. Importantly, in contrast to tocilizumab-a humanized anti-IL-6R antibody approved by the US Food and Drug Administration for the treatment of rheumatoid arthritis, juvenile idiopathic arthritis, giant cell arteritis and Castleman's disease-HZ0412a does not significantly affect the binding of IL-6 to IL-6R. Further analysis revealed that HZ0412a prevents IL-6R from binding to gp130 in vitro, while tocilizumab has a minimal effect under the same condition. Using various cell-based assays, we demonstrate that HZ0412a is noninferior to tocilizumab in inhibiting IL-6 signaling. Finally, we showed that HZ0412a is well tolerated in cynomolgus monkeys after a single subcutaneous injection at a dose of 1 or 5 mg/kg. Taken together, our results indicated that HZ0412a targets an epitope on human IL-6R that is different from that of tocilizumab, and the epitope region is essential for the interaction between IL-6R and gp130. This distinctive mode of action plus its high affinity to IL-6R led to the high potency of HZ0412a in suppressing in vitro IL-6 signaling.
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In vitro intestinal epithelium models have drawn great attention to investigating intestinal biology in recent years. However, the difficulty to maintain the normal physiological status of primary intestinal epithelium in vitro limits the applications. Here, we designed patterned electrospun polylactic acid (PLA) nanofibrous membranes with crypt-like topography and mimic ECM fibrous network to support crypt culture and construct in vitro intestinal epithelium models. The patterned electrospun PLA nanofibrous membranes modified with Matrigels at 0 °C showed high biocompatibility and promoted cell growth and proliferation. The constructed duodenum epithelium models and colon epithelium models on the patterned electrospun PLA nanofibrous membranes expressed the typical differentiation markers of intestinal epithelia and the gene expression levels were close to the original tissues, especially with the help of probiotics. The constructed intestinal epithelium models could be used to assess probiotic adhesion and colonization, which were verified to show significant differences with the Caco-2 cell models due to the different cell types. These findings provide new insights and a better understanding of the roles of biophysical, biochemical, and biological signals in the construction of in vitro intestinal epithelium models as well as the potential applications of these models in the study of host-gut microbes interactions. KEY POINTS: ⢠Patterned electrospun scaffold has crypt-like topography and ECM nanofibrous network. ⢠Matrigels at 0°C modify scaffolds more effectively than at 37°C. ⢠Synergy of biomimic scaffold and probiotics makes in vitro model close to tissue.
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Nanofibras , Alicerces Teciduais , Humanos , Engenharia Tecidual , Células CACO-2 , Diferenciação Celular , Mucosa Intestinal/metabolismo , Poliésteres/metabolismoRESUMO
Liposomes have been received much attention during the past decades as bioactive compounds carriers in food field. However, the application of liposomes is extremely limited by the structural instability during processing such as freeze-drying. In addition, the protection mechanism of lyoprotectant for liposomes during freeze-drying remains controversial. In this study, lactose, fructooligosaccharide, inulin and sucrose were used as lyoprotectants for liposomes and the physicochemical properties, structural stability and freeze-drying protection mechanism were explored. The addition of oligosaccharides could significantly suppress the changes in size and zeta potential, and the amorphous state of liposomes was negligible changed from XRD. The Tg of the four oligosaccharides, especially for sucrose (69.50 °C) and lactose (95.67 °C), revealed the freeze-dried liposomes had formed vitrification matrix, which could prevent liposomes from fusion via increasing the viscosity and reducing membrane mobility. The decrease in Tm of sucrose (147.67 °C) and lactose (181.67 °C), and the changes in functional group of phospholipid and hygroscopic capacity of lyophilized liposomes indicated oligosaccharides replaced water molecules to interact with phospholipids by hydrogen bonds. It can be concluded that the protection mechanism of sucrose and lactose as lyoprotectant was attributed to the combination of vitrification theory and water replacement hypothesis, while the water replacement hypothesis was dominated by fructooligosaccharide and inulin.
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Inulina , Lipossomos , Lipossomos/química , Inulina/química , Lactose , Fosfolipídeos/química , Oligossacarídeos , Sacarose/química , Água , LiofilizaçãoRESUMO
BACKGROUND: Chlorogenic acid (CGA), as one of the most abundant naturally occurring phenolic acids, has been documented to be beneficial for intestinal health. However, the underlying mechanism is still not fully understood. The adult intestinal stem cell is the critical driver of epithelial homeostasis and regeneration. RESULTS: This study hypothesized that CGA exerted intestinal health effects by modulating intestinal stem-cell functions. Lgr5-EGFP mice were treated for 14 days, and intestinal organoids derived from these mice were treated for 3 days, using CGA solution. In comparison with the control group, CGA treatment increased intestinal villous height and crypt depth in mice and augmented the area expansion and the number of budding intestinal organoids. Quantitative polymerase chain reaction (qPCR) analysis revealed that CGA treatment significantly increased the expression of genes coding intestinal stem-cell markers in intestinal tissue and organoids, and upregulated the expression of genes coding secretory cell lineages and enterocytes, although not statistically significantly. Fluorescence-activated cell-sorting analysis further confirmed that CGA augmented the number of stem cells. 5-Ethynyl-2'-deoxyuridine (EdU) incorporation and Ki67 immunostaining results also demonstrated that CGA treatment enhanced intestinal stem-cell proliferation. CONCLUSION: Altogether, our findings indicate that CGA could activate intestinal stem-cell and epithelial regeneration, which could contribute to the improvement of intestinal morphology or organoid growth of mice. This highlights a promising mechanism for CGA as an excellent candidate for the formulation of dietary supplements and functional foods for intestinal protection. © 2023 Society of Chemical Industry.