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This study examined the effect of Flavourzyme and Lactobacillus plantarum (L. plantarum) on protein degradation and flavor development during grass carp fermentation. The control groups comprised natural fermentation and fermentation with L. plantarum. Compared with the two control samples, those exposed to combined Flavourzyme and L. plantarum fermentation exhibited lower moisture content and enhanced protein hydrolysis, which accelerated the production of water-soluble taste substances (trichloroacetic acid-soluble peptides and free amino acids). The electronic tongue and electronic nose results indicated that the grass carp subjected to combined fermentation way displayed a more intense umami taste and aroma. Moreover, the sensory evaluation results confirmed that the combined fermentation method significantly improved the taste and odor attributes of fermented grass carp. In conclusion, combined fermentation with Flavourzyme and L. plantarum may effectively reduce fermentation time and enhance the flavor of fermented grass carp products.

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BACKGROUND: The present study aimed to investigate the cryoprotective effect of epigallocatechin gallate (EGCG) replacing sucrose on surimi during frozen storage. Substitution or partial substitution of 0.1% EGCG for sucrose (1.5%) was added to surimi, and the surimi samples without and with commercial cryoprotectants (4% sucrose and 4% sorbitol) were used as the control group. RESULTS: The results obtained suggest that, with the increase in frozen storage time, the structural performance of surimi protein gradually weakened (e.g. the decrease in the surface hydrophobicity, the increase in the total sulfhydryl and solubility, and the protein myosin heavy chain bands became shallow) and surimi gel quality gradually deteriorated (e.g. the decrease in water-holding capacity, gel strength and all texture profile attributes). However, compared with the other three group surimi samples during the frozen period, the surimi proteins with partial replacement of sucrose by EGCG had a higher total sulfhydryl group content and solubility of proteins, as well as lower surface hydrophobicity of protein, suggesting that the addition of EGCG as a partial substitute for sucrose can enhance the antifreeze ability of surimi. Meanwhile, the surimi gel with the partial replacement of sucrose by EGCG had a higher water retention capacity, gel strength and texture attributes (e.g. hardness, springiness, cohesiveness, chewiness, and resilience), indicating that the addition of EGCG as a partial substitute for sucrose can inhibit the deterioration of surimi gel quality. CONCLUSION: Overall, EGCG partially replacing sucrose can play an alternative cryoprotectant with a lower sweetness to prevent the quality of surimi from deteriorating. © 2024 Society of Chemical Industry.

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Gut microbiota (GM) is an invisible organ that plays an important role in human health. Increasing evidence suggests that polyphenols in pomegranate (punicalagin, PU) could serve as prebiotics to modulate the composition and function of GM. In turn, GM transform PU into bioactive metabolites such as ellagic acid (EA) and urolithin (Uro). In this review, the interplay between pomegranate and GM is thoroughly described by unveiling a dialog in which both actors seem to affect each other's roles. In a first dialog, the influence of bioactive compounds from pomegranate on GM is described. The second act shows how the GM biotransform pomegranate phenolics into Uro. Finally, the health benefits of Uro and that related molecular mechanism are summarized and discussed. Intake of pomegranate promotes beneficial bacteria in GM (e.g. Lactobacillus spp., Bifidobacterium spp.) while reducing the growth of harmful bacteria (e.g. Bacteroides fragilis group, Clostridia). Akkermansia muciniphila, and Gordonibacter spp., among others, biotransform PU and EA into Uro. Uro contributes to strengthening intestinal barrier and reducing inflammatory processes. Yet, Uro production varies greatly among individuals and depend on GM composition. Uro-producing bacteria and precise metabolic pathways need to be further elucidated therefore contributing to personalized and precision nutrition.

Gut microbiota plays a critical role in maintaining host health.Pomegranate is rich in bioactive components.Consumption of pomegranate positively modulates gut microbiota.Gut microbiota can transform ellagitannins in pomegranate into urolithin.Urolithin has high bioavailability and multiple health benefits.

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BACKGROUND: Ultrasound is widely used as a novel non-thermal processing technique to improve protein properties. In recent decades, applying ultrasound-assisted emulsification (UAE) to produce protein-stabilized emulsion has attracted people's attention. Instead of applying ultrasound to treat a single protein solution, UAE treatment refers to the use of sonication to a mixture of protein and oil. The purpose of this study was to compare the different effects of ultrasound treatment on the properties of myofibrillar protein (MP) in the presence or absence of soybean oil. A suitable sonication power was selected based on the change in emulsion properties. RESULTS: 300 W sonication power was selected because of its most effectively decreased emulsion droplet size and increased absolute zeta potential. Sonication more significantly increased the protein carbonyl content and disulfide bonds of the MP-soybean oil sample compared with the MP sample. Due to the presence of oil, ultrasound could unfold more protein molecules, illustrated by a lower α-helix content and intrinsic fluorescence intensity, and a higher surface hydrophobicity. Results of liquid chromatography-tandem mass spectrometry illustrated that sonication enhanced the myosin heavy chain and actin content at the soybean oil interface as well as accelerated the myosin light chain to separate from myosin in the MP-soybean oil system. CONCLUSION: Ultrasound treatment could lead to a higher level of protein oxidation and greater protein molecule exposure in the MP in the presence of oil system than in the oil-free MP system. © 2023 Society of Chemical Industry.

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NaCl is the main curing agent in dry-cured meat products, and a large amount of NaCl addition leads to high salt content of final products. Salt content and composition are important factors affecting the activity of endogenous proteases, which in turn could affect proteolysis as well as the quality of dry-cured meat products. With the increasing emphasis on the relationship between diet and health, reducing sodium content without sacrificing quality and safety of products is a great challenge for dry-cured meat industry. In this review, the change of endogenous proteases activity during processing, the potential relationship between sodium reduction strategy, endogenous proteases activity, and quality were summarized and discussed. The results showed that sodium replacement strategy and mediated-curing had a complementary advantage in influencing endogenous proteases activity. In addition, mediated-curing had the potential to salvage the negative effects of sodium substitution by affecting endogenous proteases. Based on the results, a sodium reduction strategy that sodium replacement in conjunction with mediated-curing based on endogenous proteases was proposed for the future perspective.

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The ultrasound-induced impacts on the peptide characteristics and taste of unsmoked bacon have been evaluated through the use of peptidomics and bioinformatics approaches. Furthermore, the effect of such ultrasound-induced changes on the main endogenous proteases responsible for peptide generation was also investigated. In fact, the activity of main endogenous proteases was significantly increased after ultrasonic treatment during the processing of unsmoked bacon, and contributed to an increased number and an enhanced LFQ intensity of peptides. Besides, such increased amount of peptides and LFQ intensity with up to 500 W ultrasonic treatment were beneficial for the taste improvement of the final products as shown by taste prediction analysis. Nevertheless, an excessive ultrasonic power like 750 W hindered protein hydrolysis and further exerted a negative effect on peptide generation. Therefore, ultrasound under controlled conditions could be considered as a promising way to improve the taste of unsmoked bacon.

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This study aimed to investigate the effects of nitric oxide (NO) and its induced protein S-nitrosylation on the structures and digestion properties of beef myofibrillar protein (MP). The MP was treated with 0, 50, 250, 500, and 1000 µM concentrations of NO-donor S-nitrosoglutathione (GSNO) for 30 min at 37 °C. The results indicated that GSNO treatment significantly decreased the sulfhydryl contents whereas the carbonyl contents increased. Meanwhile, compared with the control group, the surface hydrophobicity, the intrinsic fluorescence intensity, and the α-helix content of proteins were decreased significantly with the enhancement of GSNO concentrations. In addition, 250 µM GSNO treatment increased the gastric digestibility of MP, while the gastrointestinal digestibility and the release of peptides were both inhibited by 500 and 1000 µM GSNO treatments. These data demonstrate that protein S-nitrosylation can affect the in vitro digestion properties of proteins by altering the physicochemical properties and structure of MP.

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This study aimed to investigate the influence of protein oxidation on the digestive properties of beef myofibrillar protein (MP). MP was treated with a hydroxyl radical-generating system containing various concentrations of H2O2. The increased content in a free sulfhydryl group and surface hydrophobicity indicated that oxidation treatment with 1 mM H2O2 induced unfolding of MP. Reducing and nonreducing SDS-PAGE results suggested that 10 mM H2O2 oxidation treatment resulted in aggregation of MP; meanwhile, the disulfide bond was the major covalent bond involved in aggregation. Peptidomics showed that peptides in the digestion products of MP were mainly derived from myosin tail. Moderate oxidation (1 mM H2O2) facilitated the release of peptide in the rod portion (S2) of myosin, whereas excessive oxidation (10 mM H2O2) inhibited peptide release in the light meromyosin region. This work presents insightful information for the crucial impact of oxidation on meat protein digestibility from the peptidomics perspective.

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This study aimed to investigate the effect of oxidized beef protein on colon health. C57BL/6 mice were fed diets containing in vitro oxidized beef protein (carbonyl content 5.83/9.02 nmol/mg protein) or normal beef protein (control group, carbonyl content 2.27 nmol/mg protein) for 10 weeks. Histological observations showed that oxidized beef protein diet induced notable inflammatory cell infiltrations in colon. The analysis of high-throughput sequencing indicated oxidized beef protein largely altered the composition of gut microbiota (GM) by increasing proinflammatory bacteria (Desulfovibrio, Bacteroides, Enterorhabdus) while reducing beneficial bacteria (Lactobacillus, Akkermansia). In addition, oxidized beef protein remarkably increased protein fermentation in the colon, which was evidenced by the elevated i-butyrate, i-valerate, and ammonia levels in feces. Furthermore, consuming oxidized beef protein destroyed colon barrier functions by decreasing tight junction proteins expression. These changes in colonic ecosystem activated the proinflammatory pathway of lipopolysaccharide/toll-like receptor-4/nuclear factor kappa B (LPS/TLR-4/NF-κB), eventually leading to colonic inflammatory damage in mice. Taken together, these results imply that consuming oxidized beef protein detrimentally regulates GM and impairs colon health.

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Lipid oxidation and protein oxidation occur side by side in meat. Here, the effect of malondialdehyde (MDA), the major product of lipid oxidation, on the digestibility of beef myofibrillar proteins (MP) was studied. MP samples were incubated with 0, 0.1, 0.3, 0.5, and 0.7 mM MDA at 4 °C for 12 h and then subjected to in vitro gastrointestinal digestion. The result showed that MDA remarkably reduced the digestibility of MP (p < 0.05). MDA treatments significantly increased carbonyl and Schiff base contents in MP (p < 0.05). The microstructure observed by atomic force microscopy showed that MDA treatments resulted in the aggregation of MP. Non-reducing and reducing electrophoresis suggested the aggregation was mainly caused by covalent bonds including disulfide bond and carbonyl−amine bond. Proteomics analysis proved that the myosin tail was the main target of MDA attack, meanwhile, lysine residues were the major modification sites. Taken together, the above results imply that MDA induces protein oxidation, aggregation, and blockage of hydrolysis sites, consequently leading to the decrease in both gastric and gastrointestinal digestibility of MP.

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Objective To observe the anti-inflammatory effect and mechanism of bone marrow mesenchymal stem cell (BMSC) on multiple organ dysfunction syndrome (MODS) rats. Methods SD rats were randomly divided into control group, model group and BMSC group, with 10 rats in each group. Rats in model group and BMSC group were intraperitoneally injected with 7 mg/kg lipopolysaccharide (LPS) to establish MODS models. Rat BMSCs were cultured in vitro, and the fourth generation cells were used for experiments. The rats in BMSC group were injected with 1×106 BMSCs through the tail vein, and the rats in control group and model group were injected with the same amount of normal saline through the tail vein. 72 hours after MODS model was established, blood oxygen partial pressure (PaO2) and arterial carbon dioxide partial pressure (PaCO2) were detected by blood gas analyzer. ELISA were used to detect the bilirubin (TB), albumin (ALB), serum creatinine (Scr), blood urea nitrogen (BUN), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and cyclic adenosine monophosphate (cAMP). HE staining was performed to observe pathological changes in lung, liver and kidney tissues. Western blot analysis was used to detect the protein expression of protein kinase A (PKA) and nuclear factor kappa-B p65 (NF-κB p65) in tissues of lung, liver and kidney. Results Compared with control group, PaO2 in model group and BMSC group significantly decreased, while the PaCO2 significantly increased. TB, Cr, BUN, TNF-α and IL-6 levels was found increased, and ALB and cAMP levels was found decreased, along with the decreased PKA protein expression levels in lung, liver and kidney tissues, and increased NF-κB p65 protein expression levels. Compared with model group, PaO2 in BMSC group increased significantly, while its PaCO2 decreased markedly. It's also detected decreased TB, Cr, BUN, TNF-α and IL-6 levels, increased ALB and cAMP levels, as well as increased PKA protein expression levels in lung, liver and kidney tissues, and decreased NF-κB p65 protein expression levels. Conclusion BMSCs can reduce the damaged organ function and inhibit the release of inflammatory factors in MODS rats. The mechanism may be related to the change of cAMP/PKA/NF-κB signaling pathway.

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The bioactive peptides hydrolyzed from bone collagen have been found to possess health-promoting effects by regulating chronic diseases such as arthritis and hypertension. In the current study, the anti-inflammatory effect of bovine bone gelatin peptides (GP) was evaluated in 264.7 macrophages cells and followed by animal trials to investigate their interference on inflammatory cytokines and gut microbiota compositions in dextran sodium sulfate (DSS)-induced C57BL/6 mice. The GP was demonstrated to alleviate the extra secretion of interleukin-6 (IL-6), nitric oxide (NO) and tumor necrosis factor-α(TNF-α) in lipopolysaccharide (LPS)-induced RAW264.7 cells. In DSS-induced colitis mice, the gavage of GP was demonstrated to ameliorate the IBD symptoms of weight loss, hematochezia and inflammatory infiltration in intestinal tissues. In serum, the proinflammatory cytokines (TNF-α,IL-6, MCP-1, IL-1ß) were suppressed along with the decreasing effect on toll-like receptor 4 and cyclooxygenase-2 by GP treatment. In the analysis of gut microbiota, the GP was checked to modulate the abundance of Akkermansia, Parasutterella, Peptococcus, Bifidobacterium and Saccharibacteria. The above results imply that GP could attenuate DSS-induced colitis by suppressing the inflammatory cytokines and regulating the gut microbiota.

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The current study aimed to investigate the effects of ultrasound-assisted emulsification on the emulsifying and rheological properties of myofibrillar protein (MP) pork fat emulsions under different protein/fat ratios. Changes in emulsion profile, confocal laser scanning microscope images, cryo-scanning microscope images, particle size, protein solubility, surface hydrophobicity and free sulfhydryl groups were determined. Ultrasound significantly increased the emulsifying activity, the emulsifying stability and the flow index for all emulsions, while it decreased the viscosity coefficient of emulsions except for the treatment of protein/fat ratio of 1:15. The results showed that sonication reduced the particle size of the fat particles and evenly distributed the emulsion droplets. Sonication moved the distribution curve of droplet size to the smaller particle size direction and decreased the D3,2 and D4,3 values of emulsion. Sonication resulted in increased bindings between protein hydrophobic groups and fat particles. After ultrasound treatment, more sulfhydryl groups were exposed to aqueous solution, which might decrease the protein solubility in aqueous solution. Ultrasound-assisted emulsification could directly enhance the emulsifying and rheological properties of MP-stabilized pork fat emulsions at different protein/fat ratios, in particular at the ratio of 1:10.

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This study aimed to investigate theeffects of sous vide (SV), boiling (BO), and roasting (RO) on beef protein digestibility and peptide profiling in simulated gastrointestinal digestion. The results indicated that beef samples treated with SV had higher (p < 0.05) gastrointestinal digestibility (34.97%) than those treated with BO (27.59%) and RO (24.36%). Furthermore, SV (2450) resulted in more types of peptides released during gastrointestinal digestion than BO (2077) and RO (1896) and a higher proportion of 400-1200 Da peptides. The significant increase of carbonyl content, the decrease of sulfhydryl content, and the formation of covalent bonds coupled with the transformation of protein secondary structure from α-helix to ß-sheet suggested that excessive protein oxidation and aggregation occurred in BO and RO samples. These changes in BO and RO samples might explain the decreased protein digestibility and peptide release in gastrointestinal digestion. Thus, SV is a promising cooking method to improve beef protein digestibility.

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This study aimed to evaluate the effects of sous vide cooking (SV) on beef tenderness and its underlying potential mechanism. Beef semimembranosus (SM) were subjected to SV treatments at 45 °C, 55 °C and 65 °C for 4 h. Compared with control samples (CK, cooked at 75 °C until a core temperature of 72 °C was attained), SV treatment significantly promoted the release of cathepsin B and cathepsin L from lysosomes and decreased the shear force of beef SM (p < 0.05). In comparison with CK, samples treated with SV had more hydrolysis of myosin heavy chain and obtained higher myofibrillar fragmentation index, collagen solubility as well as longer sarcomere length (p < 0.05). The current study showed that the proteolysis of myofibrillar protein and collagen induced by cathepsin B and cathepsin L, and the limited longitudinal shrinkage together contributed to the improvement of beef tenderness upon SV.

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Renal cell carcinoma (RCC) has a high mortality rate among urological malignancies, and its underlying mechanisms remain unclear. Steroid receptor RNA coactivator (SRA) belongs to the long non­coding RNAs (lncRNAs) and has been demonstrated to be closely related to various types of cancer. In the present study, the decreased expression level of SRA was first confirmed in RCC tissues and cell lines by RT­qPCR. Using knockdown or overexpression systems, it was then found that SRA inhibited the proliferation of RCC cell lines and promoted their apoptosis. In addition, SRA suppressed the migration and invasion, and altered EMT­related markers in RCC cells. More importantly, it was demonstrated that SRA reduced percentage of CD44+/CD24­ cells and the sphere­forming efficiency. SRA also attenuated the expression levels of CD44, SOX­2, ABCG2 and OCT­4, which are all associated with cancer cell stemness characteristics. Although SRA increased the phosphorylation of extracellular­regulated protein kinase (ERK), the ERK1/2 pathway could not further interfere with the alteration of EMT­related markers mediated by SRA. Notably, the ERK inhibitor, PD98059, abolished ERK1/2 phosphorylation, whereas it did not exert any marked effects on cell proliferation and EMT­related markers mediated by SRA. Taken together, the findings of the present study indicate that SRA is an important molecule that inhibits the migration, invasion and stem cell characteristics of RCC cells; the ERK signaling pathway may not be involved in this process.

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Background: Homeobox B5 (HOXB5) is associated with the poor prognosis of various cancer types. However, the specific mechanism by which HOXB5 promotes the malignant progression of pancreatic cancer (PC) remains to be determined.Methods: The Cancer Genome Atlas database indicated HOXB5 expression level correlated to PC prognosis. The biological functions of HOXB5 was confirmed by colony formation, migration, and invasion assays. The effects of HOXB5 on the expression of cancer stem cell and epithelial-mesenchymal transition markers were evaluated. The downstream target of HOXB5 was miR-6723, which was detected by transcriptional assay. A xenograft tumor model was established in nude mice for the assessment of the role of HOXB5 in tumor growth and metastasis.Results: PC tissues had higher HOXB5 expression levels than noncancerous tissues, and high HOXB5 expression was significantly associated with poor PC prognosis. HOXB5 knockdown suppressed clone formation and the proliferation, invasion, and migration of PC cells in vitro. Conversely, these activities were enhanced by HOXB5 overexpression. The HOXB5 that bound two synergy motifs regulated miR-6723 expression and contributed to PC malignant progression. The role of HOXB5 in promoting tumor growth and metastasis was verified in vivo. Further investigation revealed that Twist1 and Zeb1 expression levels were increased by HOXB5.Conclusions: HOXB5 overexpression was significantly correlated with poor PC prognosis. HOXB5 accelerated the malignant progression of PC by up-regulating miR-6723, which afforded PC cells stem-like properties and facilitated the epithelial-mesenchymal transition of PC cells.

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This work investigated the influence of enzymatic tenderization on digestibility changes of beef semimembranosus proteins using peptidomics methods. Hydrolysis by proteinase K and bromelain elevated the average bitterness index of identified peptides by generating high-Q values peptides (1714-1790 Cal/mol), including KDLFDPIIQ, LIDDHFLFDKPVSPL, and QLIDDHFLFDKPVSPLLL. Proteolysis during enzymatic tenderization acted as a "pre-digestion" step and significantly elevated the degree of hydrolysis of beef protein (by 4.5-17.3%) in subsequent simulated gastrointestinal digestion. Peptidomics analysis of digests revealed large variations in the peptide composition, which was positively correlated with the degree of proteolysis during enzymatic tenderization. Enzymatic tenderization with proteinase K- (for 0.5 h) or bromelain-treated samples largely increased the survival rate (by 65.5 or 82.8%) of peptides during simulated digestion, possibly because of the "secondary enzyme-substrate interaction" effect. This work could provide a new sight into the possible influence of enzymatic tenderization on meat nutrition.

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Breast cancer (BC) is a type of malignant tumor originating from the epithelial tissue of the mammary gland, and about 20% of breast cancers are human epidermal growth factor receptor 2 positive (HER2+), which is a subtype with more aggression. Recently, HER2-positive breast cancer is often accompanied by poor prognosis of patients, and targeted therapy showed a promising prospect. To combat this disease, novel therapeutic targets are still needed. Adenylate kinase 4 (AK4) is a member of the adenylate kinase family and is expressed in the mitochondrial matrix. AK4 is involved in multiple cellular functions such as energy metabolism homeostasis. Interestingly, AK4 was observed highly expressed in several tumor tissues, and the involvement of AK4 in cancer development was generally revealed. However, the possible role of AK4 on the growth and development of breast cancer is still unclear. Here, we investigated the possible functions of AK4 on the progression of HER2-positive breast cancer. We found the high expression of AK4 in HER2-positive breast cancer tissues from patients who received surgical treatment. Additionally, AK4 expression levels were obviously correlated with clinical-pathological features, including pTNM stage (P = 0.017) and lymph node metastasis (P = 0.046). We mechanically confirmed that AK4 depletion showed the obvious impairment of cell proliferation and invasion in MCF7 and MDA-MB-231 cells. AK4 also facilitates tumor growth and metastasis of HER2-positive breast cancer in vivo. In conclusion, we identified and mechanically confirmed that AK4 is a novel therapeutic target of HER2-positive breast cancer.

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Wnt5a belongs to the large WNT family of cysteine-rich secreted glycoproteins, which is involved in multiple signaling pathways that regulate a variety of cellular processes, including cell motility, proliferation differentiation and so on during development. The regulation and signaling transduction of Wnt5a have been reported to closely relate to inflammatory response, which indicates that Wnt5a plays a critical role in the occurrence and development of inflammatory diseases. In this review, we summarized data on Wnt5a and its signaling pathway, as well as their involvement in inflammatory response. Further comprehensive understanding of the function and relationship between Wnt5a and inflammatory response would help us to develop novel diagnostic and therapeutic strategies for prevention and treatment of inflammatory diseases.

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