Reduced NO Production Research Articles

Dendropanax morbifera Extracts for Cosmetic Applications: Systematic Review and Meta-Analysis

We have conducted a systematic review and meta-analysis to evaluate the cosmetic applications of Dendropanax morbifera extracts (DMEs). A total of 261 articles were screened; however, after eliminating inappropriate studies, only 16 individual studies were eligible. The comparative standardized mean difference (SMD) between the DME treatment and control groups was used to evaluate the cosmetic properties of DME, including its biocompatibility, whitening effects, and anti-inflammatory and antimicrobial properties. DME treatment exhibited positive results in controlling hyperpigmentation, including effective inhibition of the production of tyrosinase and melanin, with SMDs of 6.85 [4.27, 9.44] and 23.38 [12.94, 33.82], respectively. Moreover, the results confirmed the anti-inflammatory properties in terms of suppressing the expression of interleukin markers (ILs) (SMD = 5.22 [3.12, 7.33]) and reducing NO production (SMD = 6.92 [2.89, 10.96]). DME treatment also effectively inhibited bacteria growth, which causes skin disorders. According to the results, DMEs are shown to be highly biocompatibility, with excellent anti-hyperpigmentation, anti-inflammatory, and antimicrobial properties that contribute significantly to improving skin appearance. The findings provide strong evidence for further research into the in vivo effects of DMEs and their potential cosmetic applications, which could lead to clinical trials in the future.

Genotoxic and Anti-Genotoxic Assessments of Fermented Houttuynia cordata Thunb. Leaf Ethanolic Extract and Its Anti-Cancer Effect in a Dual-Organ Carcinogenesis Model of Colon and Liver in Rats.

The incidence of multiple-organ cancers has recently increased due to simultaneous exposure to various environmental carcinogens. Houttuynia cordata Thunb. (H. cordata) is recognized for its many health benefits, including its anti-cancer properties. The fermentation of its leaves has been shown to significantly enhance the bioflavonoid content and its bioactivities. This study aimed to evaluate the effectiveness of fermented H.cordata leaf (FHCL) extracts against combined carcinogens and investigate the underlying mechanisms. The crude ethanolic extract of FHCL was partitioned to obtain hexane- (HEX), dichloromethane- (DCM), ethyl acetate- (ETAC), butanol- (nBA), and residue fractions. The crude ethanolic extract (200-250 μg/mL) and the DCM fraction (50 μg/mL) significantly reduced NO production in RAW264.7 macrophages. In addition, the crude extract and the DCM and ETAC fractions showed anti-genotoxicity against aflatoxin B1 and 2-amino-3,4-dimethylimidazo [4,5-f]quinoline (MeIQ) in Salmonella typhimurium assays (S9+). Despite demonstrating genotoxicity in the Salmonella mutation assay (with and without S9 activation), oral administration of the crude extract at 500 mg/kg of body weight (bw) for 40 days in rats did not induce micronucleated hepatocytes, indicating that the extract is non-genotoxic in vivo. Moreover, the crude extract significantly decreased Phase I but increased Phase II xenobiotic-metabolizing enzyme activities in the rats. Next, the anti-cancer effects of FHCL were evaluated in a dual-organ carcinogenesis model of the colon and liver in rats induced by 1,2-dimethylhydrazine (DMH) and diethylnitrosamine (DEN), respectively. The crude extract significantly reduced not only the number and size of glutathione S-transferase placental form positive foci in the liver (at doses of 100 and 500 mg/kg bw) but also the number of aberrant crypt foci in rat colons (at 500 mg/kg bw). Furthermore, FHCL significantly reduced the expression of proliferating cell nuclear antigen (PCNA) in the colon (at 100 and 500 mg/kg bw) and liver (at 500 mg/kg bw) of the treated rats. In conclusion, FHCL exhibits significant preventive properties against colon and liver cancers in this dual-organ carcinogenesis model. Its mechanisms of action may involve anti-inflammatory effects, the prevention of genotoxicity, the modulation of xenobiotic-metabolizing enzymes, and the inhibition of cancer cell proliferation. These findings support the use of FHCL as a natural supplement for preventing cancer.

Alkaloids from Piper longum L and their Anti-inflammatory Properties.

Piper longum L. (PL) is considered one of the most important species traditionally used for treating various ailments and has indicated the presence of alkaloids, flavonoids, and steroids. In this study, we isolated the chemical compounds of PL leaves, and measured NO, IL-6, iNOS, as well as COX-2 protein levels. In addition, molecular docking analysis was used to further understand the anti-inflammation effect of the compounds. We identified one new alkaloid named piperlongumine A (1) with ten known compounds (2-11). The new compound (1) and two other alkaloids 2E)-3-(4-hydroxy-3-methoxyphenyl)-1-(pyrrol-1-yl) propanone (7) and piperchabamide A (8) significantly reduced NO production in LPS-stimulated RAW 264.7 cells with the IC50 values of 0.97±0.05 μM, 0.91±0.07 μM, 1.63±0.14 μM, respectively. Moreover, at concentration of 2 μM, compound 1 inhibited approximately 98±0.64 % of IL-6 secretion, and decreased iNOS and COX-2 protein level by about 96 and 19 folds compared to LPS treatment alone, respectively. Furthermore, compounds 1, 7, and 8 were predicted to bind and inhibit IL-6, TNF-α, and iNOS, with compound 1 showing the highest binding energy of -7.09 kcal/mol. This study provides new insights for potential anti-inflammatory drug design and warrants further investigation.

Lack of AMP-activated protein kinase-α1 reduces nitric oxide synthesis in thoracic aorta perivascular adipose tissue

ObjectivePerivascular adipose tissue (PVAT) releases anti-contractile bioactive molecules including NO. PVAT anti-contractile activity is attenuated in mice lacking AMPKα1 (AMP-activated protein kinase-α1). As AMPK regulates endothelial NO synthase (eNOS) activity in cultured cells, NO synthesis was examined in PVAT from AMPKα1 knockout (KO) mice. Methods and resultsEndothelium-denuded thoracic or abdominal aortic rings were isolated from wild type (WT) and KO mice. NOS inhibition enhanced vasoconstriction in PVAT-intact thoracic aortic rings from mice of either genotype yet had no effect on abdominal rings as assessed by wire myography. Thoracic aorta PVAT exhibited increased NO production, NOS activity and levels of the brown adipose tissue marker uncoupling protein-1 (UCP1) compared to abdominal PVAT. In KO mice, NO production was significantly reduced in thoracic but not abdominal PVAT. Reduced NO production in KO thoracic PVAT was not due to altered levels or phosphorylation of eNOS but was associated with increased caveolin-1:eNOS association and caveolin-1 Tyr14 phosphorylation. A peptide that disrupts eNOS:caveolin-1 association increased NO synthesis and reduced vasoconstriction of PVAT-intact thoracic but not abdominal aortic rings. KO thoracic PVAT also exhibited reduced UCP1 levels. ConclusionsMurine thoracic aorta PVAT exhibits higher NO synthesis and UCP1 levels than abdominal aortic PVAT. Downregulation of AMPK suppresses NO synthesis which may contribute to the reduced anticontractile activity and reduced brown adipose tissue phenotype of KO thoracic PVAT. The mechanism underlying the effect of AMPK downregulation likely results from increased caveolin-1:eNOS association associated with caveolin-1 Tyr14 phosphorylation.

Prooxidant and anti-inflammatory potential of Garcinia xanthochymus fruit and its phytochemical characterisation by UHPLC-Q-Orbitrap HRMS

Pro-oxidants play a crucial role in cancer by causing oxidative stress that leads to apoptosis. The present study demonstrates the prooxidant and anti-inflammatory potential of ethyl acetate and methanolic extracts of Garcinia xanthochymus fruit. Oxidation of Trolox and NADH activity indicated the pro-oxidant capacity of the extracts. Significant decrease in cell viability in B16F10 and MDA-MB-231 cancer cell lines and significant increase in caspase 3 activity after treatment with extracts indicated pro-oxidant induced apoptosis. Pre-treatment with the extracts significantly inhibited ROS, reduced NO production, inhibited LPS-induced COX-2 and suppressed IL-6 and TNF-α expression. HRMS analysis showed the presence of compounds like biflavonoids, xanthones, phloroglucinols, benzophenones, etc. The fruit is rich in total phenolic and flavonoid contents, and have DPPH radical scavenging, ferric reducing antioxidant and metal chelating potential. This study report for the first time about the anticancer and anti-inflammatory properties of G. xanthochymus whole fruit.

Perspectives on NO X Emissions and Impacts from Ammonia Combustion Processes.

Climate change and global warming necessitate the shift toward low-emission, carbon-free fuels. Although hydrogen boasts zero carbon content and high performance, its utilization is impeded by the complexities and costs involved in liquefaction, preservation, and transportation. Ammonia has emerged as a viable alternative that offers potential as a renewable energy storage medium and supports the global economy's decarbonization. With its broader applicability in large power output applications, decentralized energy sources, and industrial locations off the grid, ammonia is increasingly regarded as an essential fuel for the future. Although ammonia provides a sustainable solution for future low-carbon energy fields, its wide-scale adoption is limited by NO X emissions and poor combustion performance under certain conditions. As research on ammonia combustion expands, recent findings reveal factors impacting the chemical reaction pathways of ammonia-based fuels, including the equivalence ratio, fuel mixture, pressure, and temperature. Investigations into ammonia combustion and NO X emissions, at both laboratory and industrial scales, have identified NO X production peaks at equivalence ratios of 0.8-0.9 for ammonia/hydrogen blends. The latest studies about the NO X emissions of the ammonia flame at different conditions and their generating pathways are reviewed in this work. Effective reduction in NO production from ammonia-based flames can be achieved with richer blends, which generate more NH i radicals. Other advanced NO X mitigation techniques such as plasma-assisted combustion have been also explored. Further research is required to address these challenges, reduce emissions, and improve efficiencies of ammonia-based fuel blends. Finally, the extinction limit of ammonia turbulent flame, its influential factors, and different strategies to promote the ammonia flame stability were discussed. The present review contributes to disseminating the latest advancements in the field of ammonia combustion and highlights the importance of refining reaction mechanisms, computational models, and understanding fundamental phenomena for practical implications.

Phytosynthesis of Silver Nanoparticles Using Mansoa alliacea (Lam.) A.H. Gentry (Bignoniaceae) Leaf Extract: Characterization and Their Biological Activities

Background. Mansoa alliacea is a native plant renowned for its medicinal properties in traditional healing in the Amazon Region. This plant is rich in polyphenols, flavonoids, anthocyanins, phenolic acids, tannins, ketones, triterpenes, as well as other bioactive compounds. Objectives. This study aims to develop an innovative, eco-friendly method for synthesizing silver nanoparticles using an aqueous extract of M. alliacea (Ma-AgNPs), enhancing the biological activities of AgNPs by leveraging the therapeutic potential of the plant’s bioactive compounds. Methods. Silver nanoparticles were synthesized using the aqueous extract of M. alliacea. The biological activities of Ma-AgNPs were assessed, including antibacterial, anti-inflammatory, antioxidant, antitumor, and anti-biofilm effects, along with evaluating their hemolytic activity. Results. Quantitative analysis revealed that Ma-AgNPs exhibit potent antibacterial activity against multidrug and non-multidrug-resistant bacteria, with MIC values ranging from 1.3 to 10.0 µg/mL. The Ma-AgNPs significantly reduced NO production by 86.9% at 4 µg/mL, indicating strong anti-inflammatory effects. They demonstrated robust antioxidant activity with an IC50 of 5.54 ± 1.48 µg/mL and minimal hemolytic activity, with no hemolysis observed up to 20 µg/mL and only 4.5% at 40 µg/mL. Their antitumor properties were notable, with IC50 values between 2.9 and 5.4 µg/mL across various cell lines, and they achieved over 50% biofilm inhibition at concentrations of 30–40 µg/mL. Conclusions. These findings underscore the potential of Ma-AgNPs for biomedical applications, particularly in developing new antimicrobial agents and bioactive coatings with reduced toxicity. This research highlights a sustainable approach that not only preserves but also amplifies the inherent biological activities of plant extracts, paving the way for innovative therapeutic solutions.

Inhibitory Effects of Naringenin on LPS-Induced Skin Inflammation by NF-κB Regulation in Human Dermal Fibroblasts.

Flavonoids are important natural compounds characterized by their extensive biological activities. Citrus flavonoids represent a significant segment of the broader flavonoid category. Naringenin, an integral part of this series, is recognized for its powerful anti-inflammatory and antioxidant properties. In addition, considering the lack of existing research on naringenin's potential effectiveness and intracellular mechanisms of action in skin-related applications, especially as a cosmetic ingredient, this study aimed to explore naringenin's role in reducing the fundamental generation of reactive oxygen species. This was achieved by examining its inhibitory effects on the expression levels of NADPH oxidase and iNOS, ultimately leading to a reduction in NO production. This research examined the anti-inflammatory and antioxidant capacities of naringenin by employing a cellular senescence model of LPS-induced HDFs. The evaluation of naringenin's efficacy was validated through several investigative procedures, including the NF-κB luciferase assay, ELISA assay, and qRT-PCR. To verify the anti-inflammatory effectiveness of naringenin, we measured the responsive elements of NF-κB using a luciferase reporter assay. This assessment revealed that naringenin could decrease the concentration of genes activated by NF-κB. Moreover, we found that naringenin inhibited the transcriptional expression of known NF-κB-regulated inflammatory cytokines, including IL-1β, IL-6, and IL-8. In addition, results from the qRT-PCR analysis indicated that naringenin facilitated a reduction in iNOS expression. Based on the data gathered and analyzed in this study, it can be conclusively inferred that naringenin possesses promising potential as a cosmetic ingredient, offering both anti-inflammatory and antioxidant benefits.

Three anti-inflammatory polysaccharides from Lonicera japonica Thunb.: Insights into the structure-function relationships

This study demonstrates the feasibility of producing three polysaccharides (neutral LJP-1, acidic LJP-2 and acidic LJP-3) with significant in vitro and in vivo anti-inflammatory activities from the flowers of Lonicera japonica. The three polysaccharides differed in chemical composition, molecular weight distribution, glycosidic linkage pattern, functional groups and morphology. They exhibited excellent protective effects (in a dose-dependent manner) in lipopolysaccharide-injured RAW264.7 macrophages and CuSO4-damaged zebrafish via reducing NO production and inhibiting the overexpressions of inflammation-related transcription factors, inflammatory proteins and cytokines in the NF-κB/MAPK signaling pathways. Their anti-inflammatory effects varied owing to their different molecular characteristics and chemical compositions. Overall, LJP-2 at 400μg/mL was the most effective. LJP-2 consisted mainly of → 5)-α-L-Araf (1→, →4)-α-L-GalpA (1→ and →2)-α-L-Rhap (1→ residues with terminal T-β-D-Glcp. Thus, honeysuckle flowers are good sources of anti-inflammatory polysaccharides, and precise fractionation enables the production of potent anti-inflammatory agents for the development of functional foods and healthcare products.

Anti-inflammatory activity and structural analysis of fucoidan extracted from Sargassum fusiforme via ESI-TOF MS spectrometry

This study used Celluclast-assisted hydrolysis and DEAE-cellulose anion-exchange chromatography to extract, purify, and assess the anti-inflammatory activity of fucoidan derived from Sargassum fusiforme. The Celluclast-assisted hydrolysate was used for obtaining ethanol-precipitated S. fusiforme fucoidans (SFF) which were rich in polysaccharides and sulfates. Five fractions obtained through DEAE-cellulose chromatography, notably SFF-F5, displayed high sulfate content (29.16 ± 0.15%). The anti-inflammatory potential of these fractions was tested in lipopolysaccharide-induced RAW 264.7 macrophages. SFF-F5 was found to significantly reduce NO production at non-toxic concentrations of 12.5, 25, and 50 μg/mL. Structural analysis using Matrix assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS) revealed repetitive patterns in the fucoidan fractions. And Electrospray ionization mass spectrometry (ESI-TOF MS) spectrum of SFF fractions identified various structures of glycans bound with fucose and sulfate (Fuc1(SO3)1, Fuc2(SO3)1, Hex5Fuc1(SO3)1). In-silico evaluation based on the detected glycan structures indicated that fucoidan fractions could inhibit the dimerization of the TLR4-MD2 complex, demonstrating their potential as anti-inflammatory agents. This result confirm that the anti-inflammatory activity varies depending on the glycan composition of fucoidan. This study provides a foundation for further exploration into the relationship between the structure and anti-inflammatory activity of S. fusiforme-derived fucoidan, facilitating its effective industrial application.

Millingtonia hortensis L.f. ethanol extract exerts in vivo and in vitro anti-inflammatory activities through inhibition of Syk kinase in NF-κB pathway

Ethnopharmacological relevanceMillingtonia hortensis L.f., commonly known as tree jasmine or Indian cork tree, is native to South Asia and Southeast Asia. Traditionally, its stem bark, leaves, and roots are employed for pulmonary, gastrointestinal, and antimicrobial purposes, while the flowers are used in treating asthma and sinusitis.Aim of the study: The underlying anti-inflammatory mechanisms of M. hortensis remain relatively unexplored. Therefore, we studied the anti-inflammatory effects of M. hortensis and the molecular mechanisms of its ethanol extracts (Mh-EE) both in vitro and in vivo. Materials and methodsNitric oxide (NO) production was assessed using Griess reagent, while cell viability of RAW264.7 cell and HEK293T cells were determined via the MTT assay. Constituent analysis of Mh-EE using GC/MS-MS and HPLC, and mRNA expression of inflammatory cytokines was measured through PCR and RT-PCR. Protein levels were analyzed using western blotting. The thermal stability of Mh-EE was evaluated by CESTA. Lastly, a gastritis in vivo model was induced by HCl/EtOH, and protein expression levels were measured using western blotting. ResultsMh-EE significantly reduced NO production in LPS-induced RAW264.7 cells without substantially affecting cell viability. Additionally, Mh-EE decreased the expression of proinflammatory factors, such as iNOS, IL-1β and COX2. Furthermore, Mh-EE downregulated TLR4 expression, altered MyD88 recruitment, and suppressed phosphorylation of Syk, IKKα, IκBα and AKT. Simultaneously, Mh-EE also attenuated NF-κB signaling in HCl/EtOH-induced mice. ConclusionsMh-EE exerts anti-inflammatory effects by suppressing p-Syk in the NF-κB pathway, and it has potential as a novel treatment agent for inflammatory diseases.

Flame characteristics and NO emission behaviors of methane/ammonia counterflow premixed flames with opposed N2 or air stream

Behaviors of methane/ammonia counterflow premixed flames and NO emission are numerically and experimentally investigated with opposed N2 or air stream by varying strain rate (ag) and equivalence ratio (ϕ). The premixed stream is 80% methane and 20% ammonia by volume mixed with air considering the utilization of ammonia by blending hydrocarbon fuels. The opposed stream is either N2 or air considering the potential application in gas turbines having single- or two-stage combustion. The simulations are conducted by increasing ag up to flame extinction for a specified ϕ and an experiment is conducted to evaluate various kinetic mechanisms. The extinction strain rate increases monotonically with ϕ with opposed air stream (AS), while increases and then decreases with opposed nitrogen stream (NS). The extinction mechanisms are explained by adopting the concepts of the local equilibrium temperature and heat loss ratio. For lean and rich mixtures with NS and lean mixtures with AS, the extinction occurs due to comparable effects of incomplete reaction and heat loss from the flame to the downstream burnt region. While for rich mixtures with AS, the extinction is attributed mainly to incomplete reaction. The contribution of thermal NO as compared with prompt NO in NO production is weak for all cases with NS and AS. The emission index (EINO) for lean and rich mixtures with NS and for lean mixtures with AS decreases monotonically with the increase of strain rate for all ϕ. Increasing equivalence ratio at a specified ag decreases NO production for lean and rich mixtures with NS and lean mixtures with AS. While for rich mixtures with AS, the EINO increases monotonically with ag. From the reaction pathway analysis, for lean mixtures, the effect of increasing ag on the reduction of NO production is attributed by weakening NH-related reaction paths, while NH2-related reaction paths change little. When ϕ increases for a specified ag with NS, NO production is reduced by weakening NH2-related reaction paths and strengthening NH-related reaction paths. For rich mixtures with AS, increasing ag increases NO production by strengthening NH2-related reaction paths.

Endothelial cell HDAC1 mediates aortic stiffness in diet-induced obese mice

Histone deacetylase 1 (HDAC1) is an important regulator of cell differentiation and is associated with progression of cardiovascular disease (CVD). Our lab previously showed that over-expression of HDAC1 in cultured endothelial cells reduces NO production. Other investigators have shown increased HDAC1 protein in models of CVD. However, there is a gap in knowledge of whether endothelial-specific HDAC1 mediates vascular dysfunction in models of CVD. Diet induced obesity (DIO) is a leading risk factor for CVD, thus our studies are focused on a chronic high fat feeding mouse model. Our studies utilize a mouse model of DIO where mice are fed a normal diet (ND, 10% fat) or high fat diet (DIO, 45% fat) beginning at 8 weeks of age for 20 weeks. Previously, we showed that this model of DIO in male mice significantly increases aortic stiffness (measured by pulse wave velocity, PWV) and increases aortic thickness compared to ND (PWV: unpaired T test, n=9-12, normal diet (ND) vs DIO, p=<0.0001; Aortic Thickness: unpaired T test, n=6, ND vs DIO, p=0.005). We hypothesized that endothelial-specific HDAC1 mediates aortic dysfunction in a mouse model of DIO. We generated inducible endothelial-Scl-Cre-ER-HDAC1 knockout (KO) mice (iEC-HDAC1KO), which specifically deletes HDAC1 in endothelial cells after tamoxifen injection, and the flox control mice (iEC-HDAC1flox), which does not delete HDAC1 after tamoxifen injection. We assessed aortic stiffness of iEC-HDAC1 KO and iEC-HDAC1flox in ND and DIO mice. We treated with tamoxifen for 5 days when ND mice were 20 weeks old and measured PWV 2 weeks later. Interestingly, iEC-HDAC1KO mice had significantly reduced PWV compared to iEC-HDAC1flox mice on ND (unpaired T test; n=4-5, p=0.009). At week 18 of the DIO protocol, mice were given daily tamoxifen injections for 5 days. After week 20 of the DIO protocol, iEC-HDAC1KO DIO mice had significantly reduced PWV compared to iEC-HDAC1flox DIO mice (unpaired T test; n=4-5; p=0.008). These data indicate that endothelial-specific HDAC1 mediates vascular stiffness and remodeling in high fat fed mice. Further investigation is needed to understand the relationship of endothelial HDAC1 and NO in the context of DIO and aortic health. Funding: R01 DK134562, F31HL167626. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A Characterization of Biological Activities and Bioactive Phenolics from the Non-Volatile Fraction of the Edible and Medicinal Halophyte Sea Fennel (Crithmum maritimum L.).

Although the biochemical composition and biological properties of the volatile fraction of the halophyte sea fennel (Crithmum maritimum L.) have been largely described, little is known about its polar constituents and bioactivities. Here, a hydromethanolic extract of Crithmum maritimum (L.) leaves was fractionated, and the fractions were evaluated in vitro for antioxidant (using DPPH, ABTS, and FRAP bioassays), anti-inflammatory (inhibition of NO production in RAW 264.7 macrophages), antidiabetic (alpha-glucosidase inhibition), neuroprotective (inhibition of acetylcholinesterase), and skin-protective (tyrosinase and melanogenesis inhibitions) activities. Polar fractions of the extract were rich in phenolics and, correlatively, displayed a strong antioxidant power. Moreover, fractions eluted with MeOH20 and MeOH80 exhibited a marked inhibition of alpha-glucosidase (IC50 = 0.02 and 0.04 mg/mL, respectively), MeOH60 fractions showed a strong capacity to reduce NO production in macrophages (IC50 = 6.4 μg/mL), and MeOH80 and MeOH100 fractions had strong anti-tyrosinase activities (630 mgKAE/gDW). NMR analyses revealed the predominance of chlorogenic acid in MeOH20 fractions, 3,5-dicaffeoylquinic acid in MeOH40 fractions, and 3-O-rutinoside, 3-O-glucoside, 3-O-galactoside, and 3-O-robinobioside derivatives of quercetin in MeOH60 fractions. These compounds likely account for the strong antidiabetic, antioxidant, and anti-inflammatory properties of sea-fennel polar extract, respectively. Overall, our results make sea fennel a valuable source of medicinal or nutraceutical agents to prevent diabetes, inflammation processes, and oxidative damage.

Anti-inflammatory potential via the MAPK signaling pathway of Lactobacillus spp. isolated from canine feces.

Two probiotic candidates, Lactobacillus reuteri C1 (C1) and Lactobacillus acidophilus C5 (C5), which were previously isolated from canines, were evaluated in the present study. L. reuteri and L. acidophilus have anti-oxidant, anti-inflammatory, immune-enhancing, and anti-cancer properties and exhibit various probiotic effects in humans and animals. The strains C1 and C5 demonstrated good tolerance to acid and bile salt exposure, exhibited effective adhesion to HT-29 cell monolayer, and displayed sensitivity to antibiotics, thus affirming their probiotic characteristics. Moreover, C1 and C5 exhibited the ability to downregulate the expression of inducible NO synthase (iNOS), an immunomodulatory factor, leading to a reduction in NO production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. These strains also demonstrated potent anti-inflammatory effects in LPS-stimulated RAW 264.7 cells, achieved through the augmentation of anti-inflammatory cytokine IL-10 expression and the inhibition of pro-inflammatory cytokine IL-1β expression. These anti-inflammatory effects of C1 and C5 were closely associated with the mitogen-activated protein kinase (MAPK) signaling pathway. The results of the present study suggest that the C1 and C5 probiotic candidates attenuate LPS-induced inflammation via the MAPK signaling pathway and the strains can be used as probiotics considering their anti-inflammatory potential.

Numerical investigation on the effects of air-staged strategy and ammonia co-firing ratios on NO emission characteristics using the Conjugate heat transfer method

Ammonia co-firing is recognized as a promising approach within the realm of achieving carbon neutrality. This method not only effectively mitigates carbon emissions but also augments the stability of ammonia combustion. However, achieving low NO emissions in ammonia combustors remains a formidable challenge. The present study employed numerical simulations to assess the impact of an air-staged strategy and varying ammonia co-firing ratios on NO emissions within a 10-kW industrial ammonia co-firing furnace. To accurately represent the thermal conditions at the furnace walls, a Conjugate Heat Transfer (CHT) model encompassing both solid materials and reacting flow regions was utilized. Initially, the numerical results were compared with experimentally measured NO emissions, validating the accuracy of the customized solver. Subsequently, the effectiveness of the air-staged strategy under the parallel injection system was verified. This approach created a fuel-rich region near the burner primary nozzle, significantly reducing NO production. Under different ammonia co-firing ratios, with ammonia injection levels increasing from 0% to 100%, NO emissions at the furnace outlet displayed an initial increase, followed by a decrease, peaking at a co-firing ratio of 50%. Further analysis of the Rate of Production (ROP) of NO within the furnace revealed that at higher ammonia co-firing ratios, ammonia serves both as a reducing agent and as a fuel for heat release. These findings provide valuable insights for the development and application of ammonia-based industrial furnaces.

The Antioxidant and Anti-Inflammatory Properties of Wild Bilberry Fruit Extracts Embedded in Mesoporous Silica-Type Supports: A Stability Study.

Polyphenolic extracts from wild bilberries (Vaccinium myrtillus L.) have shown antioxidant and anti-inflammatory effects, but they are prone to degradation when exposed to environmental factors, limiting their use in biomedical applications. To overcome this issue, this study proposed the embedding of wild bilberry fruit ethanolic extracts in pristine mesoporous silica functionalized with organic groups (mercaptopropyl and propionic acid), as well as coated with fucoidan, a biopolymer. Herein, we report a stability study of free and incorporated extracts in mesoporous silica-type supports in high-humidity atmospheres at 40 °C up to 28 days, using HPLC analysis, thermal analysis, and radical scavenging activity determination. Better chemical and thermal stability over time was observed when the extracts were incorporated in mesoporous silica-type supports. After 12 months of storage, higher values of antioxidant activity were determined for the extract embedded in the supports, silica modified with mercaptopropyl groups (MCM-SH), and fucoidan-coated silica (MCM-SH-Fuc) than that of the free extract due to a synergistic activity between the support and extract. All encapsulated extracts demonstrated remarkable effects in reducing NO production in LPS-stimulated RAW 264.7 cells. The treatment with extract embedded in MCM-SH-Fuc in a dose of 10 μg/mL surpassed the effect of free extract in the same concentration. For the extract encapsulated in an MCM-SH support, a lower IC50 value (0.69 μg/mL) towards COX-2 was obtained, comparable with that of Indomethacin (0.6 μg/mL). Also, this sample showed a higher selectivity index (2.71) for COX-2 than the reference anti-inflammatory drug (0.98). The developed formulations with antioxidant and anti-inflammatory properties could be further used in nutraceuticals.

Effectiveness of Ya-Samarn-Phlae in diabetic wound healing: Evidence from in vitro studies and a multicenter randomized controlled clinical trial

Ethnopharmacological relevanceYa-Samarn-Phlae (YaSP) has traditionally been widely used in southern Thailand for treating chronic and infected wounds, including diabetic foot ulcers. However, there are only a limited number of clinical studies supporting the use of this polyherbal formulation. Therefore, the present work aims to provide clinical evidence to support the application of YaSP, prepared according to a standardized traditional procedure (T-YaSP). Additionally, its potential chemical markers and wound healing-related biological activities were examined. Materials and methodsThe in vitro wound healing-related biological activities of YaSP ethanol extract and T-YaSP, including antibacterial activity against Staphylococcus epidermidis, inhibition and eradication of staphylococcal biofilm, anti-inflammatory effects, and enhancement of human dermal fibroblast migration in scratch wounds, were examined using well-established protocols. The chemical profiles of the ethanol extract of YaSP and T-YaSP were compared, and with promising chemical markers, arecoline, alpha-mangostin, and curcumin were selected and quantified using the HPLC method. A prospective, multicenter, randomized, controlled, parallel-group study was conducted over 12 weeks to evaluate the efficacy of the YaSP solution as an adjunct therapy, combined with standard wound care, for diabetic ulcers compared to standard treatment. ResultsThe YaSP extract reduces NO production and can scavenge NO radicals in LPS-induced RAW 264.7 macrophage cells. Additionally, in a scratch assay, this extract and one of its herbal components, Curcuma longa, enhance the migration of human dermal fibroblasts. T-YaSP, containing 2.412 ± 0.002 mg/g of arecoline, 2.399 ± 0.005 mg/g of curcumin, and 0.017 ± 0.000 mg/g of α-mangostin, has shown the ability to inhibit the development and eradicate the mature biofilm of S. epidermidis. The use of T-YaSP as an adjunct therapy led to a significantly higher proportion of patients achieving healing within six weeks compared to the standard treatment group (36%/9 patients vs. 4%/1 patient; p = 0.013). After 12 weeks, 19 out of 25 patients in the T-YaSP group experienced complete healing, whereas only four patients in the standard treatment group achieved complete wound healing (76% in the T-YaSP group vs. 16% in the control group; p < 0.001). ConclusionThe results presented here represent the first randomized controlled trial to demonstrate the effectiveness of the traditional polyherbal solution, T-YaSP, which exhibits a wide range of wound healing-related activities. Utilizing T-YaSP as an adjunctive treatment resulted in a significant improvement in the number of type 2 diabetic patients achieving complete healing. However, to explore and utilize YaSP further, conducting a double-blind, randomized controlled trial with a larger population is necessary.

Role of the CCL5 and Its Receptor, CCR5, in the Genesis of Aldosterone-Induced Hypertension, Vascular Dysfunction, and End-Organ Damage.

Aldosterone has been described to initiate cardiovascular diseases by triggering exacerbated sterile vascular inflammation. The functions of CCL5 (C-C motif chemokine ligand 5) and its receptor CCR5 (C-C motif chemokine receptor 5) are well known in infectious diseases, their contributions to aldosterone-induced vascular injury and hypertension remain unknown. We analyzed the vascular profile, blood pressure, and renal damage in wild-type (CCR5+/+) and CCR5 knockout (CCR5-/-) mice treated with aldosterone (600 µg/kg per day for 14 days) while receiving 1% saline to drink. Vascular function was analyzed in aorta and mesenteric arteries, blood pressure was measured by telemetry and renal injury and inflammation were analyzed via histology and flow cytometry. Endothelial cells were used to study the molecular signaling whereby CCL5 induces endothelial dysfunction. Aldosterone treatment resulted in exaggerated CCL5 circulating levels and vascular CCR5 expression in CCR5+/+ mice accompanied by endothelial dysfunction, hypertension, and renal inflammation and damage. CCR5-/- mice were protected from these aldosterone-induced effects. Mechanistically, we demonstrated that CCL5 increased NOX1 (NADPH oxidase 1) expression, reactive oxygen species formation, NFκB (nuclear factor kappa B) activation, and inflammation and reduced NO production in isolated endothelial cells. These effects were abolished by antagonizing CCR5 with Maraviroc. Finally, aorta incubated with CCL5 displayed severe endothelial dysfunction, which is prevented by blocking NOX1, NFκB, or CCR5. Our data demonstrate that CCL5/CCR5, through activation of NFκB and NOX1, is critically involved in aldosterone-induced vascular and renal damage and hypertension placing CCL5 and CCR5 as potential therapeutic targets for conditions characterized by aldosterone excess.

Roles and Mechanisms of Dopamine Receptor Signaling in Catecholamine Excess Induced Endothelial Dysfunctions.

Endothelial dysfunction may contribute to pathogenesis of Takotsubo cardiomyopathy, but mechanism underlying endothelial dysfunction in the setting of catecholamine excess has not been clarified. The study reports that D1/D5 dopamine receptor signaling and small conductance calcium-activated potassium channels contribute to high concentration catecholamine induced endothelial cell dysfunction. For mimicking catecholamine excess, 100 μM epinephrine (Epi) was used to treat human cardiac microvascular endothelial cells. Patch clamp, FACS, ELISA, PCR, western blot and immunostaining analyses were performed in the study. Epi enhanced small conductance calcium-activated potassium channel current (ISK1-3) without influencing the channel expression and the effect was attenuated by D1/D5 receptor blocker. D1/D5 agonists mimicked the Epi effect, suggesting involvement of D1/D5 receptors in Epi effects. The enhancement of ISK1-3 caused by D1/D5 activation involved roles of PKA, ROS and NADPH oxidases. Activation of D1/D5 and SK1-3 channels caused a hyperpolarization, reduced NO production and increased ROS production. The NO reduction was membrane potential independent, while ROS production was increased by the hyperpolarization. ROS (H2O2) suppressed NO production. The study demonstrates that high concentration catecholamine can activate D1/D5 and SK1-3 channels through NADPH-ROS and PKA signaling and reduce NO production, which may facilitate vasoconstriction in the setting of catecholamine excess.