Enhancement of Immune Functions by Limosilactobacillus reuteri KBL346: In Vitro and In Vivo Studies.

Lactobacilli have been widely used as probiotics because of their benefits for intestinal health and physiological functions. Among a variety of Lactobacillus genera, Limosilactobacillus reuteri has been studied for its ability to exert anti-inflammatory functions and its role in controlling metabolic disorders, as well as the production of the antimicrobial compound reuterin. However, the effects and mechanisms of L. reuteri on enhancing immune responses in the immunosuppressed states have been relatively understudied. In this study, we isolated an immunomodulatory strain, namely, L. reuteri KBL346 (KBL346), from a fecal sample of a 3-month-old infant in Korea. We evaluated the immunostimulatory activity and hematopoietic function of KBL346 in macrophages and cyclophosphamide (CPA)-induced immunosuppressed mice. KBL346 increased the phagocytic activity against Candida albicans MYA-4788 in macrophages, and as biomarkers for this, increased secretions of nitric oxide (NO) and prostaglandin E2 (PGE2) were confirmed. Also, the secretions of innate cytokines (TNF-α, IL-1ß, and IL-6) were increased. In CPA-induced immunosuppressed mice, KBL346 at a dosage of 1010 CFU/kg protected against spleen injury and suppressed levels of immune-associated parameters, including NK cell activity, T and B lymphocyte proliferation, CD4+ and CD8+ T cell abundance, cytokines, and immunoglobulins in vivo. The effects were comparable or superior to those in the Korean red ginseng positive control group. Furthermore, the safety assessment of KBL346 as a probiotic was conducted by evaluating its antibiotic resistance, hemolytic activity, cytotoxicity, and metabolic characteristics. This study demonstrated the efficacy and safety of KBL346, which could potentially be used as a supplement to enhance the immune system.

Antibacterial Photodynamic Inactivation of Fagopyrin F from Tartary Buckwheat (Fagopyrum tataricum) Flower against Streptococcus mutans and Its Biofilm.

The objective of this study was to determine reactive oxygen species (ROS) produced by fagopyrin F-rich fraction (FFF) separated from Tartary buckwheat flower extract exposed to lights and to investigate its antibacterial photodynamic inactivation (PDI) against Streptococcus mutans and its biofilm. ROS producing mechanisms involving FFF with light exposure were determined using a spectrophotometer and a fluorometer. S. mutans and its biofilm inactivation after PDI treatment of FFF using blue light (BL; 450 nm) were determined by plate count method and crystal violet assay, respectively. The biofilm destruction by ROS produced from FFF after exposure to BL was visualized using confocal laser scanning microscopy (CLSM) and field emission scanning electron microscope (FE-SEM). BL among 3 light sources produced type 1 ROS the most when applying FFF as a photosensitizer. FFF exposed to BL (5 and 10 J/cm2) significantly more inhibited S. mutans viability and biofilm formation than FFF without the light exposure (p < 0.05). In the PDI of FFF exposed to BL (10 J/cm2), an apparent destruction of S. mutans and its biofilm were observed by the CLSM and FE-SEM. Antibacterial PDI effect of FFF was determined for the first time in this study.

Flavonoids in common and tartary buckwheat hull extracts and antioxidant activity of the extracts against lipids in mayonnaise.

Buckwheat hulls, generally discarded as waste, have been known to possess various flavonoids and high antioxidant activities. The objective of this study was to determine effect of extracting solvents [water, ethanol (20%, 50%, 80%, and 100%), methanol, and acetone] on total phenolic content, flavonoid content and composition, and antioxidant activities of common and tartary buckwheat hull extracts. Antioxidative effect of common and tartary buckwheat hull extracts on lipids in mayonnaise was also investigated. Vitexin, isovitexin, isoorientin, orientin, rutin, isoquercetin, and quercetin were identified in the common buckwheat hull extracts, while rutin, quercetin, isoorientin, and isoquercetin were in the tartary buckwheat hull extracts. The methanol and 80% ethanol extracts had more flavonoids than the others, while the aqueous ethanol extracts from both of the hulls had more total phenolics and antioxidant activities. Oxidative stability of lipids in mayonnaises added with common and tartary buckwheat hull extracts (0.02 and 0.08%, w/w) prepared by 50% ethanol were higher than that in the mayonnaise with butylated hydroxytoluene (0.02%) and control. Oxidative stability was not significantly different between the mayonnaises added with the two buckwheat hull extracts.

Anti-inflammatory activities of black raspberry seed ellagitannins and their structural effects on the stimulation of glucagon-like peptide-1 secretion and intestinal bitter taste receptors.

This study aimed to investigate the anti-inflammatory effects of ellagitannins from black raspberry seeds (BS) in vivo and the structural effects of ellagitannins on glucagon-like peptide-1 (GLP-1) secretion and intestinal bitter taste receptor (TAS2R) stimulation. For animal study, BS ellagitannin fraction (BSEF) was orally administered to mice with colitis induced by dextran sulfate sodium (DSS). The BSEF supplementation alleviated colonic inflammation, regulated inflammation-related cytokine levels in the mice with colitis, and increased the total GLP-1 secretion and GLP-1 receptor mRNA level in the inflamed gut. It also augmented the colonic gene expressions of mouse TAS2R (mTAS2R) 108, 119, 126, 131, 138, and 140; meanwhile, only mTAS2R108 expression was downregulated by DSS treatment. Six BS ellagitannins (sanguiin H-6, casuarictin, pedunculagin, acutissimin A, castalagin, and vescalagin) induced GLP-1 secretion in STC-1 cells and upregulated mTAS2R108, 119, 126, and 138 gene expressions. The major ellagitannins in BS (sanguiin H-6, casuarictin, pedunculagin, and acutissimin A) upregulated the gene expressions of mTAS2R131 and/or 140 known to be specifically distributed in mouse colon. Through molecular docking with mTAS2R108, the hexahydroxydiphenoyl, flavan-3-ol, glucose, and nonahydroxytriphenoyl moieties of the six BS ellagitannins were predicted to be involved in interacting with the receptor. BS ellagitannins could be promising candidates for preventing colon inflammation, likely via GLP-1 secretion induced by intestine-specific TAS2Rs.

Profiling of phenolic acids, flavonoids, terpenoids, and steroid derivatives in coconut (Cocos nucifera L.) haustorium.

Coconut haustorium (CH) is formed inside coconut shell during coconut germination. This study aimed to investigate the compositions and contents of CH phytochemicals. Phytochemical compositions and contents in CH were analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and spectrophotometrical method. Five phenolic acids and four flavonoids were identified in CH. Ferulic acid and myricetin were the most abundant among phenolic acids and flavonoids identified in CH, respectively. Nepetariaside and 1-methylene-5α-androstan-3α-ol-17-one glucuronide were the most abundant terpenoids and steroid derivatives identified in CH, respectively. To our knowledge, this study screened several classes of phytochemicals in CH for the first. Terpenoids and steroid derivatives were likely to be more major phytochemicals than phenolic acids and flavonoids in CH. The functionality of CH itself and the compounds found in CH might be utilized in functional foods or cosmetics. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01300-6.

Effects of black raspberry extract on gut microbiota, microbial metabolites, and expressions of the genes involved in cholesterol and bile acid metabolisms in rats fed excessive choline with a high-fat diet.

In our previous study, black raspberry (BR) reduced the serum levels of trimethylamine-N-oxide and cholesterol in rats fed excessive choline with a high-fat diet (HFC). We hypothesized that gut microbiota could play a crucial role in the production of trimethylamine and microbial metabolites, and BR could influence gut microbial composition. This study aimed to elucidate the role of BR on changes in gut microbiota and microbial metabolites in the rats. The phylogenetic diversity of gut microbiota was reduced in the rats fed HFC, while that in the BR-fed group was restored. The BR supplementation enriched Bifidobacterium and reduced Clostridium cluster XIVa. In the BR-fed group, most cecal bile acids and hippuric acid increased, while serum lithocholic acid was reduced. The BR supplementation upregulated Cyp7a1 and downregulated Srebf2. These results suggest that BR extract may change gut bacterial community, modulate bile acids, and regulate gene expression toward reducing cholesterol. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01267-4.

Distinct Changes in Microbiota-Mediated Intestinal Metabolites and Immune Responses Induced by Different Antibiotics.

The cocktails of antibiotics are utilized to study the functions of microbiota. There have been studies on the alteration of not only the microbiota composition but also the host's metabolism or immunity. However, the bacterial species associated with these altered physiologic markers are still unclear. Therefore, we supplied mice with drinking water containing ampicillin (AMP), vancomycin (VAN), neomycin (NEO), or metronidazole (MET) to observe the effect of each antibiotic on helper T cells and inflammation-related gene expression and metabolism, including amino acid metabolism and changes in gut microbiota. We observed major changes in gut microbiota in mice treated with AMP and VAN, respectively, immediately after administration. The abundance of the genera Parabacteroides and Akkermansia increased in the AMP and VAN groups, while Prevotella almost disappeared from both groups. The compositional changes in intestinal metabolites in the AMP and VAN groups were more distinct than those in the NEO and MET groups, which was similar to the microbiome results. In particular, the most distinct changes were observed in amino acid related metabolism in AMP and VAN groups; the amounts of phenylalanine and tyrosine were increased in the AMP group while those were decreased in the VAN group. The changed amounts of intestinal amino acids in each of the AMP and VAN groups were correlated with increases in the abundance of the genera Parabacteroides and Akkermansia in the AMP and VAN groups, respectively. The most distinctive changes in intestinal gene expression were observed in the ileum, especially the expression Th17-related genes such as rorgt, il17a, and il17f, which decreased dramatically in the guts of most of the antibiotic-treated groups. These changes were also associated with a significant decrease in Prevotella in both the AMP and VAN groups. Taken together, these findings indicate that changes in gut microbiota as well as host physiology, including host metabolism and immunity, differ depending on the types of antibiotics, and the antibiotic-induced gut microbiota alteration has a correlation with host physiology such as host metabolic or immunological status. Thus, the immune and metabolic status of the host should be taken into account when administering antibiotics.

Black raspberry extract can lower serum LDL cholesterol via modulation of gut microbial composition and serum bile acid profile in rats fed trimethylamine-N-oxide with a high-fat diet.

Blood trimethylamine-N-oxide (TMAO) has been associated with cardiovascular disease. Black raspberry (Rubus occidentalis, BR) has been regarded to be beneficial for cardiovascular health. This study aimed to investigate how BR extract affects serum lipid profile, gut microbial composition, metabolites in rats fed TMAO with a high-fat diet. Dietary TMAO increased serum LDL cholesterol, while BR extract decreased its level. α-Diversity of gut microbiota was not changed; however, in the rats fed TMAO, Macellibacteroides and Mucispirillum were enriched, while Ruminococcaceae was reduced. The BR supplementation could restore Macellibacteroides, Clostridium, and Ruminococcaceae. The BR supplementation increased cecal hippuric acid and serum farnesoid X receptor-antagonistic bile acids, including ursodeoxycholic acid (UDCA), tauro-α-muricholic acid, and tauro-UDCA. The BR supplementation tended to upregulate Cyp7a1 and Abcg5 expressions while downregulating Srebf2 and Hmgcr expressions. BR extract affects the gut bacterial community and microbial metabolites, lowering serum LDL cholesterol in rats with elevated serum TMAO. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01079-y.

Co-administration of Lactobacillus gasseri KBL697 and tumor necrosis factor-alpha inhibitor infliximab improves colitis in mice.

Inflammatory bowel disease (IBD) refers to disorders involving chronic inflammation of the gastrointestinal tract. Well-established treatments for IBD have not yet to be suggested. To address this gap, we investigated the effects of co-administration of Lactobacillus gasseri (L. gasseri) KBL697 and infliximab (IFX), the first approved tumor necrosis factor (TNF)-alpha inhibitor, on the dextran sodium sulfate-induced colitis mouse model. 2 × 109 colony-forming units/g of L. gasseri KBL697 were administered to seven-week-old female C57BL/6J mice daily by oral gavage. On day three, IFX (5 mg/kg) suspended in 1 × PBS (200 µL) was intravenously injected in the IFX-treated group and all mice were sacrificed on day nine. Co-administration of L. gasseri KBL697 and IFX improved colitis symptoms in mice, including body weight, disease activity index, colon length, and histology score. Additionally, pro-inflammatory cytokines, such as interferon-gamma, interleukin (IL)-2, IL-6, IL-17A, and TNF were significantly decreased, while IL-10, an anti-inflammatory cytokine, was increased. Expression levels of tight junction genes and CD4 + CD25 + Foxp3 + T regulatory cells in the mesenteric lymph nodes were synergistically upregulated with the combined treatment. Furthermore, co-administered mice displayed altered cecum microbial diversity and composition with increases in the genus Prevotella. Related changes in the predicted amino and nucleic acid metabolic pathways were also evident, along with increased acetate and butyrate level. Therefore, the synergistic effect of L. gasseri KBL697 and IFX co-administration is a possible method of prevention and treatment for IBD.

Lactobacillus acidophilus KBL409 Reduces Kidney Fibrosis via Immune Modulatory Effects in Mice with Chronic Kidney Disease.

SCOPE: Intestinal dysbiosis has been reported to play an important role in the pathogenesis of various diseases, including chronic kidney disease (CKD). Here, to evaluate whether probiotic supplements can have protective effects against kidney injury in an animal model of CKD is aimed. METHODS AND RESULTS: An animal model of CKD is established by feeding C57BL/6 mice a diet containing 0.2% adenine. These model mice are administered Lactobacillus acidophilus KBL409 daily for 4 weeks. Features of adenine-induce CKD (Ade-CKD) mice, such as prominent kidney fibrosis and higher levels of serum creatinine and albuminuria are improved by administration of KBL409. Ade-CKD mice also exhibit a disrupted intestinal barrier and elevate levels of TNF-α, IL-6, and 8-hydroxy-2'-deoxyguanosine. These changes are attenuated by KBL409. Administration of KBL409 significantly reduces macrophage infiltration and promotes a switch to the M2 macrophage phenotype and increasing regulatory T cells. Notably, the NLRP3 inflammasome pathway is activated in the kidneys of Ade-CKD and decreases by KBL409. In primary kidney tubular epithelial cells treated with p-cresyl sulfate, short-chain fatty acids significantly increase M2 macrophage polarization factors and decrease profibrotic markers. CONCLUSIONS: These results demonstrate that supplementation with the probiotic KBL409 has beneficial immunomodulating effects and protects against kidney injury.

Akkermansia muciniphila secretes a glucagon-like peptide-1-inducing protein that improves glucose homeostasis and ameliorates metabolic disease in mice.

The gut microbiota, which includes Akkermansia muciniphila, is known to modulate energy metabolism, glucose tolerance, immune system maturation and function in humans1-4. Although A. muciniphila is correlated with metabolic diseases and its beneficial causal effects were reported on host metabolism5-8, the molecular mechanisms involved have not been identified. Here, we report that A. muciniphila increases thermogenesis and glucagon-like peptide-1 (GLP-1) secretion in high-fat-diet (HFD)-induced C57BL/6J mice by induction of uncoupling protein 1 in brown adipose tissue and systemic GLP-1 secretion. We apply fast protein liquid chromatography and liquid chromatography coupled to mass spectrophotometry analysis to identify an 84 kDa protein, named P9, that is secreted by A. muciniphila. Using L cells and mice fed on an HFD, we show that purified P9 alone is sufficient to induce GLP-1 secretion and brown adipose tissue thermogenesis. Using ligand-receptor capture analysis, we find that P9 interacts with intercellular adhesion molecule 2 (ICAM-2). Interleukin-6 deficiency abrogates the effects of P9 in glucose homeostasis and downregulates ICAM-2 expression. Our results show that the interactions between P9 and ICAM-2 could be targeted by therapeutics for metabolic diseases.

Protective Effects of Black Raspberry (Rubus occidentalis) Extract against Hypercholesterolemia and Hepatic Inflammation in Rats Fed High-Fat and High-Choline Diets.

Choline is converted to trimethylamine by gut microbiota and further oxidized to trimethylamine-N-oxide (TMAO) by hepatic flavin monooxygenases. Positive correlation between TMAO and chronic diseases has been reported. Polyphenols in black raspberry (BR), especially anthocyanins, possess various biological activities. The objective of this study was to determine the effects of BR extract on the level of choline-derived metabolites, serum lipid profile, and inflammation markers in rats fed high-fat and high-choline diets. Forty female Sprague-Dawley (SD) rats were randomly divided into four groups and fed for 8 weeks as follows: CON (AIN-93G diet), HF (high-fat diet), HFC (HF + 1.5% choline water), and HFCB (HFC + 0.6% BR extract). Serum levels of TMAO, total cholesterol, and low-density lipoprotein (LDL)-cholesterol and cecal trimethylamine (TMA) level were significantly higher in the HFC than in the HFCB. BR extract decreased mRNA expression of pro-inflammatory genes including nuclear factor-κB (NF-κB), interleukin (IL)-1ß, IL-6, and cyclooxygenase-2 (COX-2), and protein expression of NF-κB and COX-2 in liver tissue. These results suggest that consistent intake of BR extract might alleviate hypercholesterolemia and hepatic inflammation induced by excessive choline with a high-fat diet via lowering elevated levels of cecal TMA and serum TMAO in rats.

Roseburia spp. Abundance Associates with Alcohol Consumption in Humans and Its Administration Ameliorates Alcoholic Fatty Liver in Mice.

Although a link between the gut microbiota and alcohol-related liver diseases (ALDs) has previously been suggested, the causative effects of specific taxa and their functions have not been fully investigated to date. Here, we analyze the gut microbiota of 410 fecal samples from 212 Korean twins by using the Alcohol Use Disorders Identification Test (AUDIT) scales to adjust for host genetics. This analysis revealed a strong association between low AUDIT scores and the abundance of the butyrate-producing genus Roseburia. When Roseburia spp. are administered to ALD murine models, both hepatic steatosis and inflammation significantly improve regardless of bacterial viability. Specifically, the flagellin of R. intestinalis, possibly through Toll-like receptor 5 (TLR5) recognition, recovers gut barrier integrity through upregulation of the tight junction protein Occludin and helps to restore the gut microbiota through elevated expression of IL-22 and REG3γ. Our study demonstrates that Roseburia spp. improve the gut ecosystem and prevent leaky gut, leading to ameliorated ALDs.

Lactobacillus fermentum species ameliorate dextran sulfate sodium-induced colitis by regulating the immune response and altering gut microbiota.

We evaluated immunometabolic functions of novel Lactobacillus fermentum strains (KBL374 and KBL375) isolated from feces of healthy Koreans. The levels of inflammatory cytokines, such as interleukin (IL)-2, interferon-γ, IL-4, IL-13, and IL-17A, were decreased, and that of the anti-inflammatory cytokine IL-10 was increased, in human peripheral blood mononuclear cells (PBMCs) treated with the L. fermentum KBL374 or KBL375 strain. When these strains were orally administered to mice with dextran sulfate sodium (DSS)-induced colitis, both L. fermentum KBL374 and KBL375 showed beneficial effects on body weight, disease activity index score, colon length, cecal weight, and histological scores. Furthermore, both L. fermentum KBL374 and KBL375 modulated the innate immune response by improving gut barrier function and reducing leukocyte infiltration. Consistent with the PBMC data, both L. fermentum KBL374- and KBL375-treated DSS mice demonstrated decreased Th1-, Th2-, and Th17-related cytokine levels and increased IL-10 in the colon compared with the DSS control mice. Administration of L. fermentum KBL374 or KBL375 to mice increased the CD4+CD25+Foxp3+Treg cell population in mesenteric lymph nodes. Additionally, L. fermentum KBL374 or KBL375 administration reshaped and increased the diversity of the gut microbiota. In particular, L. fermentum KBL375 increased the abundance of beneficial microorganisms, such as Lactobacillus spp. and Akkermansia spp. Both L. fermentum KBL374 and KBL375 may alleviate inflammatory diseases, such as inflammatory bowel disease, in the gut by regulating immune responses and altering the composition of gut microbiota.

Black Raspberry Seed Oil Improves Lipid Metabolism by Inhibiting Lipogenesis and Promoting Fatty-Acid Oxidation in High-Fat Diet-Induced Obese Mice and db/db Mice.

The objective of this study was to evaluate the beneficial effect of α-linolenic acid-rich black raspberry seed (BRS) oil on lipid metabolism in high-fat diet (HFD)-induced obese and db/db mice. Five-week-old C57BL/6 mice were fed diets consisting of 50% calories from lard, 5% from soybean, and 5% from corn oil (HFD), or 50% calories from lard and 10% from BRS oil (HFD + BRS oil diet) for 12 weeks. Six-week-old C57BL/KsJ-db/db mice were fed diets consisting of 16% calories from soybean oil (standard diet), 8% from soybean, and 8% from BRS oil, or 16% from BRS oil for 10 weeks. The BRS oil diets lowered the levels of triacylglycerol, nonesterified fatty acids, and total cholesterol in serum and liver of both of the obese and db/db mice as compared with the HFD and standard diet, respectively. mRNA levels of lipogenesis markers including cluster of differentiation 36, fatty-acid-binding protein 1, sterol regulatory element binding protein 1c, fatty-acid synthase, and solute carrier family 25 member 1 in the liver of the BRS oil groups were lower than those in the liver of the HFD and standard groups in the obese and db/db mice, respectively. On the other hand, fatty-acid oxidation markers including carnitine palmitoyltransferase 1A, acyl-CoA dehydrogenase, hydroxylacyl-CoA dehydrogenase α, and acyl-CoA oxidase in the liver of the BRS oil groups were higher than those in the liver of the HFD and standard groups in the obese and db/db mice, respectively. Peroxisome proliferator-activated receptor α mRNA and protein levels increased in the liver and epididymal adipose tissue of the obese and db/db mice fed BRS oil compared with HFD and standard diet, respectively. BRS oil might improve lipid metabolism by inhibiting lipogenesis and promoting fatty-acid oxidation in HFD-induced obese and db/db mice.

Physicochemical characteristics and anti-oxidant activities of farm-cultivated and mountain-cultivated ginseng seeds.

Physicochemical characteristics and anti-oxidant capacities of seeds from two farm-cultivated and one mountain-cultivated ginsengs (Panax ginseng Meyer) (MG) were determined. The seeds had 17.9-22.1% (dry basis) crude lipids, 11.5-15.2% crude proteins, and 1.4-1.7% ash. Oleic acid (77.9-78.5%) was the predominant fatty acid in the seed oils, followed by linoleic acid (16.6-17.4%). The highest Hunter b value, carotenoids, (ß + γ)-tocotrienol, and δ-tocotrienol, and the lowest α-tocotrienol were observed in the seed oils from MG. Squalene was also the most abundant in the MG seed oils. ß-Sitosterol was the major phytosterol in the seed oils with MG the highest. Defatted seed meal extracts from MG possessed the most total phenolics and flavonoids, and the highest DPPH and ABTS radical scavenging activities. These results suggest that MG seeds may be a novel source of functional foods.

Dietary Black Raspberry Seed Oil Ameliorates Inflammatory Activities in db/db Mice.

The objective of this study was to evaluate the status of the markers related to inflammation in db/db mice fed black raspberry seed (BRS) oil, which is rich in α-linolenic acid. Mice were divided into four groups: (1) C57BL/6 mice fed 16 % calories from soybean oil (normal CON); (2) C57BL/KsJ-db/db mice fed 16 % calories from soybean oil (CON); (3) C57BL/KsJ-db/db mice fed 8 % calories from soybean and 8 % calories from BRS oil (BRS 50 %); and (4) C57BL/KsJ-db/db mice fed 16 % calories from BRS oil (BRS 100 %). After 10 weeks, n-6/n-3 fatty acid ratios were significantly (P < 0.05) lower in the livers and epididymal adipose tissues of the BRS 50 % and BRS 100 % mice than in the CON. Serum TNFα and IL-6 were significantly (P < 0.05) lower in the BRS 50 % and BRS 100 % than in the CON. Serum IL-10 was significantly (P < 0.05) higher in the BRS 100 % than the CON. In the liver and epididymal adipose tissue, mRNA levels of pro-inflammatory markers in the BRS 50 % and BRS 100 % were lower than in the CON. Anti-inflammatory markers were higher in the epididymal adipose tissues of the BRS 50 % and BRS 100 % than in the CON. In the epididymal adipose tissue, macrophage infiltration markers (F4/80 and CD68) and leptin mRNA were significantly (P < 0.05) lower in the BRS 50 % and BRS 100 % than in the CON. Results of this study suggest that BRS oil may have anti-inflammatory effects in obese diabetic mice by ameliorating inflammatory responses.