Mode of Action of Antimicrobial Potential Protease SH21 Derived from Bacillus siamensis.

Global public health is facing a major issue with emerging resistance to antimicrobial agents. Antimicrobial agents that are currently on the market are strong and efficient, but it has not been ruled out that these medications will eventually cause resistance to bacteria. Exploring novel bioactive compounds derived from natural sources is therefore, crucial to meet future demands. The present study evaluated the mode of action of the antimicrobial potential protease enzyme SH21. Protease SH21 exhibited antimicrobial activity, strong heat stability (up to 100 °C), and pH stability (pH 3.0 to 9.0). In terms of mode of action, we found that protease SH21 was able to disrupt the bacterial cell membrane as the results of the nucleotide leakage and cell membrane permeability assay. In addition, we also checked inner membrane permeability by PI uptake assay which suggested that protease SH21 has the ability to enter the bacterial cell membrane. Our results revealed that the antimicrobial protease SH21 might be a promising candidate for treating microbial infections.

Attenuation of Oxidative Damage via Upregulating Nrf2/HO-1 Signaling Pathway by Protease SH21 with Exerting Anti-Inflammatory and Anticancer Properties In Vitro.

Oxidative damage and inflammation are among the very significant aspects interrelated with cancer and other degenerative diseases. In this study, we investigated the biological activities of a 25 kDa protease (SH21) that was purified from Bacillus siamensis. SH21 exhibited very powerful antioxidant and reactive oxygen species (ROS) generation inhibition activity in a dose-dependent approach. The mRNA and protein levels of antioxidant enzymes such as superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione peroxidase 1 (GPx-1) were enhanced in the SH21-treated sample. SH21 also increased the transcriptional and translational activities of NF-E2-related factor 2 (Nrf2) with the subsequent development of detoxifying enzyme heme oxygenase-1 (HO-1). In addition, SH21 showed potential anti-inflammatory activity via inhibition of nitric oxide (NO) and proinflammatory cytokines, such as TNF-α, IL-6, and IL-1ß, production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. At concentrations of 60, 80, and 100 µg/mL, SH21 potentially suppressed nitric oxide synthase (iNOS) and cytokine gene expressions. Furthermore, SH21 significantly released lactate dehydrogenase (LDH) enzyme in cancer cell supernatant in a concentration-dependent manner and showed strong activity against three tested cancer cell lines, including HL-60, A549, and Hela. Our results suggest that SH21 has effective antioxidant, anti-inflammatory, and anticancer effects and could be an excellent therapeutic agent against inflammation-related diseases.

Synergistic Effect, Improved Cell Selectivity, and Elucidating the Action Mechanism of Antimicrobial Peptide YS12.

Antimicrobial peptides (AMPs) have attracted considerable attention as potential substitutes for traditional antibiotics. In our previous research, a novel antimicrobial peptide YS12 derived from the Bacillus velezensis strain showed broad-spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria. In this study, the fractional inhibitory concentration index (FICI) indicated that combining YS12 with commercial antibiotics produced a synergistic effect. Following these findings, the combination of YS12 with an antibiotic resulted in a faster killing effect against bacterial strains compared to the treatment with the peptide YS12 or antibiotic alone. The peptide YS12 maintained its antimicrobial activity under different physiological salts (Na+, Mg2+, and Fe3+). Most importantly, YS12 exhibited no cytotoxicity towards Raw 264.7 cells and showed low hemolytic activity, whereas positive control melittin indicated extremely high toxicity. In terms of mode of action, we found that peptide YS12 was able to bind with LPS through electrostatic interaction. The results from fluorescent measurement revealed that peptide YS12 damaged the integrity of the bacterial membrane. Confocal laser microscopy further confirmed that the localization of peptide YS12 was almost in the cytoplasm of the cells. Peptide YS12 also exhibited anti-inflammatory activity by reducing the release of LPS-induced pro-inflammatory mediators such as TNF-α, IL-1ß, and NO. Collectively, these properties strongly suggest that the antimicrobial peptide YS12 may be a promising candidate for treating microbial infections and inflammation.

Biochemical Characterization and Application of a Detergent Stable, Antimicrobial and Antibiofilm Potential Protease from Bacillus siamensis.

Proteases are important enzymes that are engaged in a variety of essential physiological functions and have a significant possible use in industrial applications. In this work, we reported the purification and biochemical characterization of a detergent stable, antimicrobial, and antibiofilm potential protease (SH21) produced by Bacillus siamensis CSB55 isolated from Korean fermented vegetable kimchi. SH21 was purified to obtain homogeneity via ammonium sulfate precipitation (40-80%), Sepharose CL-6B, and Sephadex G-75 column. By analyzing the SDS-PAGE and zymogram, it was determined that the molecular weight was around 25 kDa. The enzyme activity was almost completely inhibited in the presence of PMSF and DFP, which indicated that it was a member of the serine protease family. SH21 showed excellent activity with a broad range of pH and temperature, with its maximum pH of 9.0 and temperature of 55 °C. The enzyme had estimated Km and Vmax values of 0.197 mg/mL and 1.22 × 103 U/mg, respectively. In addition, it preserved good activity in the presence of different organic solvents, surfactants, and other reagents. This enzyme showed good antimicrobial activity that was evaluated by MIC against several pathogenic bacteria. Furthermore, it exhibited strong antibiofilm activity as determined by MBIC and MBEC assay and degraded the biofilms, which were analyzed by confocal microscopic study. These properties established that SH21 is a potent alkaline protease that can be used in industrial and therapeutic applications.

A novel antimicrobial peptide YS12 isolated from Bacillus velezensis CBSYS12 exerts anti-biofilm properties against drug-resistant bacteria.

Nowadays, the abuse of antibiotics has led to the rise of multi-drug-resistant bacteria. Antimicrobial peptides (AMPs), with broad-spectrum antimicrobial activity have attracted considerable attention as possible alternatives to traditional antibiotics. In this work, we aimed to evaluate the antimicrobial and anti-biofilm activity of an antimicrobial peptide designed as YS12 derived from Bacillus velezensis CBSYS12. The strain CBSYS12 was isolated from Korean food kimchi and purified followed by ultrafiltration and sequential chromatographic methodology. Hereafter, Tricine SDS-PAGE revealed a single protein band of around 3.3 kDa that was further confirmed in situ inhibitory activity of the gel. A similar molecular weight (~ 3348.4 Da) protein also appeared in MALDI-TOF confirming the purity and homogeneity of peptide YS12. Intriguingly, YS12 revealed a strong antimicrobial activity with a minimum inhibitory concentration (MIC) value ranging from 6 to 12 µg/ml for both Gram-positive and Gram-negative bacteria, such as E. coli, P. aeruginosa, MRSA 4-5, VRE 82, and M. smegmatis. We also determined the mode of action of the peptide against pathogenic microorganisms using different fluorescent dyes. In addition, the anti-biofilm assay demonstrated that peptide YS12 was able to inhibit biofilm formation  around 80% for both bacterial strains E. coli and P. aeruginosa at 80 µg/ml. Notably, YS12 exhibited a greater biofilm eradication activity than commercial antibiotics. In summary, our study proposed that peptide YS12 may be used as a promising therapeutic agent to overcome drug and biofilm-related infections.

Activation of Nrf2/HO-1 by Peptide YD1 Attenuates Inflammatory Symptoms through Suppression of TLR4/MYyD88/NF-κB Signaling Cascade.

In this study, we investigate the immunomodulatory effects of a novel antimicrobial peptide, YD1, isolated from Kimchi, in both in vitro and in vivo models. We establish that YD1 exerts its anti-inflammatory effects via up-regulation of the Nrf2 pathway, resulting in the production of HO-1, which suppresses activation of the NF-κB pathway, including the subsequent proinflammatory cytokines IL-1ß, IL-6, and TNF-α. We also found that YD1 robustly suppresses nitric oxide (NO) and prostaglandin E2 (PGE2) production by down-regulating the expression of the upstream genes, iNOS and COX-2, acting as a strong antioxidant. Collectively, YD1 exhibits vigorous anti-inflammatory and antioxidant activity, presenting it as an interesting potential therapeutic agent.

Reduction of Oxidative Stress through Activating the Nrf2 mediated HO-1 Antioxidant Efficacy Signaling Pathway by MS15, an Antimicrobial Peptide from Bacillus velezensis.

The efficient culture and purification of antimicrobial peptides (AMPs), along with intense antioxidant activity, have drawn the interest to study antioxidant activity mechanism. We report the culture conditions optimization, efficient biosynthesis, and purification of an antioxidant peptide MS15 from Bacillus velezensis obtained from fermented food that would generate heme oxygenase-1 (HO-1) expression and lead to nuclear factor erythroid 2-related factor-2 (Nrf2) nuclear translocation. We explored the ability of kinetics and potency for the bacterial killing to work against various pathogenic bacteria. A bioassay showed the lysis zone of MS15 by tricine SDS-PAGE near at 6 kDa. MALDI-TOF/MS verified molecular weight, and the existence of a molecular mass of 6091 Da was reported by purity. The MIC of MS15 ranged from 2.5-160 µg/mL for many pathogenic bacteria, showing greater potency. In macrophage RAW 264.7 cells, MS15 was exposed to assess its inhibitory effect against the generation of reactive oxygen species (ROS) in oxidative stress. In the sample treated group, the translation, and transcriptional levels of CAT (catalase), GPx (glutathione peroxidase), and SOD (superoxide dismutase) were significantly greater. In short, MS15 has significant antioxidant properties, reducing ROS production in RAW 264.7 cells, and raising the translation and transcriptional rates of antioxidant enzymes with stimulating HO-1 induction facilitated by Nrf2.

A Novel Peptide Oligomer of Bacitracin Induces M1 Macrophage Polarization by Facilitating Ca2+ Influx.

Antimicrobial peptides (AMPs) are components of the innate immune system and form the first defense against pathogens for various organisms. In the present study, we assessed whether CSP32, a novel AMP oligomer of bacitracin isolated from a strain of Bacillus spp., regulates the polarization of murine macrophage-like RAW 264.7 cells. CSP32 stimulated phagocytosis while inducing the appearance of the typical M1 polarized macrophage phenotype; these M1 macrophages play a role in host defense against pathogens. Furthermore, our results showed that CSP32 enhanced the expression and production of pro-inflammatory mediators, such as cytokines and chemokines. In addition, the CSP32-stimulated inflammatory mediators were induced mainly by the mitogen-activated protein kinase/nuclear factor kappa B (MAPK/NF-κB) signaling pathway during M1 macrophage polarization. In particular, CSP32 markedly increased the numbers of Ca2+-positive macrophages while upregulating phospholipase C and activating protein kinase Cε. Furthermore, the inhibition of intracellular Ca2+ by BAPTA-AM, a Ca2+ chelator, significantly suppressed the CSP32-mediated phagocytosis, inflammatory mediator production, and NF-κB activation. In conclusion, our data suggested that CSP32-stimulated M1 macrophage polarization is dependent on the calcium signaling pathway and may result in enhanced immune capacities.

Industrial attributes of ß-glucanase produced by Bacillus sp. CSB55 and its potential application as bio-industrial catalyst.

The ß-glucanase produced from Bacillus sp. CSB55 not only depicts the potent industrial characteristics but also relates as bio-industrial catalyst supporting the spontaneous formation of the products, high hydrolytic efficiency, and feasibility of the enzymatic reaction. A homogeneous ß-glucanase (GluB55) was purified via various purification processes resulting in 11.69% yield and 14.24-fold purity. Biochemical characterization of the purified enzyme revealed the molecular mass of approximately 40 kDa, which was verified by zymography. The optimum activity of GluB55 was determined at pH 7.2 and 55 °C. GluB55 could highly hydrolyze carboxymethylcellulose and was stable over a wide range of pH, retaining more than 70% residual activity at pH 5.8-11.0 and carried 100% thermostability as high as 60 °C. In addition, it showed 68% residual activity at 70 °C. The N-terminal amino acid sequence of GluB55 was Ala-Asn-Pro-Glu-Leu-Val-Asn-X-Gln-Ala-X-X-Ala-X-Gln-Gly. The enzyme activity was stimulated by Co2+ (158.6%), Zn2+ (211.1%), Mn2+ (264.4%), and Ba2+ (211.4%). Enzyme kinetics showed Km and Vmax values of 0.022 mg mL-1 and 994.56 ± 3.72 U mg-1, respectively. Q10 was calculated to be 1.12. ∆H, ∆G, and ∆S were low revealing that the formation of the transition phase and conversion to the product is very well organized. The lower the free energy change (∆G), the more feasible is the reaction.

Comparative Analysis Of Anticholinergics Prescribed To Elderly Patients At A Korean Long-Term Care Facility According To Beers Criteria 2003, 2012, And 2015 And Anticholinergic-Burden Rating Scales: A Cross-Sectional Retrospective Study.

BACKGROUND: The Beers Criteria is used as a reference to identify potentially inappropriate medications (PIMs) prescribed to older people, and anticholinergic risk measurement scales (ARMSs) have been continuously made for measuring the anticholinergic burden. This study aimed to evaluate the concordance between any anticholinergics among PIMs identified by the Beers Criteria and those assessed by 9 different ARMSs. METHODS: This study was retrospectively conducted with Korean older patients hospitalized in the long-term care facility between March 2014 and August 2015. The data were collected through the chart review of electronic medical records of the patients. The Beers Criteria 2003, 2012, and 2015 were used to detect PIMs, and the following ARMSs were also employed to assess their potential anticholinergic effects: Anticholinergic Cognitive Burden Scale (2008), Anticholinergic Risk Scale (2008), Chew's Scale (2008), Anticholinergic Drug Scale (ADS; 2006), Anticholinergic Activity Scale (AAS; 2010), Anticholinergic Load Scale (2011), Clinician-Rated Anticholinergic Scale (2008), Duran's Scale (2013), and Anticholinergic Burden Classification (2006). RESULTS: The eligible patients who met inclusion and exclusion criteria were 216 during the study period. Most patients were females (70.4%), and the mean age was 81.0 ± 6.7 years. Approximately 70%, 86%, and 87% of the patients included were identified as using at least one PIM according to the Beers Criteria 2003, 2012, and 2015, respectively. Compared with the Beers Criteria 2003, the versions of 2012 and 2015 showed more improved concordance associated with the ARMSs. When the Beers Criteria 2015 was compared with the ARMSs, the lowest concordance was found for AAS (κ = 0.153; 95% CI, 0.079-0.227), whereas the highest concordance was observed for ADS (κ = 0.530; 95% CI, 0.406-0.654). CONCLUSION: The heterogeneity between the Beers Criteria and the ARMSs was observed. Compared with the Beers Criteria 2003, the versions of 2012 and 2015 showed more enhanced concordance associated with the ARMSs.

Production of Novel Polygalacturonase from Bacillus paralicheniformis CBS32 and Application to Depolymerization of Ramie Fiber.

Polygalacturonase (EC. 3.2.1.15) is an enzyme that hydrolyzes the alpha-1,4 glycosidic bonds between galacturonic acid. In this study, an alkaline polygalacturonase producer Bacillus paralicheniformis CBS32 was isolated from kimchi (conventional Korean fermented food). The 16S rRNA sequence analysis of the isolated strain revealed that it was 99.92% identical to B. paralicheniformis KJ 16LBMN01000156. The polygalacturonase from B. paralicheniformis CBS32 was named PN32, and the purified PN32 showed a 16.8% yield and a 33-fold purity compared to the crude broth. The molecular mass, 110 kDa, was determined by SDS-PAGE, and the active band was confirmed by zymography analysis. The N-terminal amino acid sequence residues of PN32 were determined to be Gly-Val-Lys-Glu-Val-X-Gln-Thr-Phe. In the sequence comparison, PN32 was suggested as a novel polygalacturonase, since the sequence was not matched with the previous reports. In an application study, enzymatic depolymerization of ramie was performed for fiber degumming, and the result showed that the PN32 had a 28% higher depolymerization compared to the commercial pectinase. Overall, based on the results, PN32 has high potential for industrial applications.

Immobilization of an alkaline endopolygalacturonase purified from Bacillus paralicheniformis exhibits bioscouring of cotton fabrics.

Pectin degrading enzyme has been increasing interest in an industrial application as biocatalysts, such as juice, textile, and wine industry. Bacillus paralicheniformis CBS3, isolated from popular traditional Korean food (kimchi), produced a novel extracellular thermostable alkaline endopolygalacturonase (BPN3). In this study, BPN3 was purified to 22.04-fold with a recovery yield of 18.93% and specific activity of 2216.41 U/mg by gel filtration and anion exchange column chromatography. The molecular mass of BPN3 was approximately 53 kDa as analyzed by SDS-PAGE and pectic zymography. The N-terminal sequence of BPN3 was AIPVILAX. BPN3 was stable over a broad pH range (8.14-11.47), was thermally stable at 50-60 °C, and functioned optimally in pH 9.1 at 60 °C. BPN3 had Km and Vmax values of 0.039 mg/mL and 747.9 ± 1.2 U mg- 1, respectively, whereas pectin from apple as substrate. BPN3 activity was remarkably affected by metal ions, modulators, and detergents. Digalacturonic acid (GA2) was the major oligosaccharide produced by hydrolysis of BPN3. Immobilized BPN3 was active over a pH range (8.1-11.5), temperature (50-60)°C, and remained stable with 63.34 and 43.41% of its relative activity during second and third cycle, respectively. Desized cotton exhibited highest reducing sugar liberation through optimized conditions of bioscouring. Bioscouring effectiveness of BPN3 was characterized by the comparison of weight loss for purified BPN3 with commercial pectinase and comparison of BPN3 with grey fabric. BPN3 was simple to purify, had high thermal stability, and was stable over a broad pH range that suggests its suitability for bioscouring application as an industrial catalyst.

Anti-inflammatory effect of Allium hookeri on carrageenan-induced air pouch mouse model.

Inflammation is a commonly observed immune reaction, and rheumatoid arthritis is a particularly severe inflammatory disease. In this study, we used an air pouch mouse model to evaluate the anti-inflammatory potential of Allium hookeri, which has both been used as a culinary material and a traditional medicine in south-eastern Asia for many years. Allium hookeri suppressed typical symptoms of inflammation, such as condensation of the air pouch membrane, and inhibited the expression of several inducible proinflammatory cytokines such as IL-1ß, IL-6, IL-13, and TNF-α. In order to determine the molecules modulating the inflammatory effect of carrageenan treatment, the components in Allium hookeri were analyzed by GC-MS, and linoleic acid, which have anti-inflammatory effect, was detected. From the results, we concluded that the anti-inflammatory effect of Allium hookeri might be attributed to linoleic acid, which could be promising candidates for anti-inflammatory drugs that have no adverse effects.

Opuntia humifusa modulates morphological changes characteristic of asthma via IL-4 and IL-13 in an asthma murine model.

Asthma is a chronic pulmonary disease that affects an estimated 235 million people worldwide, but asthma drugs have many adverse effects. Opuntia humifusa (eastern prickly pear) has been used as a food and traditional medicine worldwide; however, its anti-asthmatic effects have not been reported. We evaluated O. humifusa as a potential therapeutic or preventive component of anti-asthmatic drugs. We divided ovalbumin-sensitized mice into the following groups: normal control, asthma-induced control, dexamethasone-treated group (positive control), 50 mg/kg O. humifusa-treated group, 100 mg/kg O. humifusa-treated group, and 500 mg/kg O. humifusa-treated group. Levels of Th1/Th2/Th17-related cytokines were evaluated using RT-PCR, ELISA, and immunohistochemistry. O. humifusa dose-dependently suppressed the morphological changes typically observed in asthma, such as goblet cell hyperplasia, inflammatory cell infiltration, mucous hypersecretion, and relative basement membrane thickening in the respiratory system. These results may be attributable to regulation of Th1-/Th2-/Th17-related factors, especially interleukin (IL)-4 and IL-13. We conclude that O. humifusa is a potential anti-asthmatic functional food. Abbreviations: O. humifusa: Opuntia humifusa; Th: helper T; RT-PCR: real-time polymerase chain reaction; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; WHO: World Health Organization; IFN-γ: interferon gamma; TNF-α: tumor necrosis factor-alpha; IgE: immunoglobulin E; CD: cluster of differentiation; OVA: ovalbumin; DEX: dexamethasone; BALF: bronchoalveolar fluid; H&E: hematoxylin and eosin; PAS: periodic acid-schiff; PBS: phosphate-buffered saline; BM: basement membrane; cDNA: complementary deoxyribonucleic acid; RNA: ribo nucleic acid; RIPA: radioimmunoprecipitation assay; IHC: immunohistochemistry; HPLC: high-performance liquid chromatography; SD: standard deviation; WBC: white blood cells; APCs: antigen-presenting cells.

Anticancer activity and antioxidant potential of Aponogeton undulatus against Ehrlich ascites carcinoma cells in Swiss albino mice.

The in vivo anticancer and in vitro antioxidant effects of the crude methanol extract of Aponogeton undulatus, in addition to its various organic fractions, were investigated. Various assay methods, including the total antioxidant capacity assay, lipid peroxidation inhibition assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay and ferrous reducing power assessment were used to evaluate the antioxidant potential of the crude extract and its organic fractions. The ethyl acetate fraction (EAU) demonstrated the highest antioxidant capacity (175.80±0.41 mg/g equivalent ascorbic acid), which exhibited IC50 values of DPPH scavenging activity and maximum lipid peroxidation of 38.84±0.02 and 42.52±0.32 µg/ml, respectively. The reducing power of the extract was concentration dependent. The in vivo antitumor activity was evaluated against Ehrlich ascites carcinoma (EAC) cell-bearing Swiss albino mice. EAUs (50, 100 and 200 mg/kg body weight) were administered for nine consecutive days. On day 10, half of the mice were sacrificed and the remaining mice were evaluated for lifespan. Hematological profiles including, red blood cell, white blood cell and hemoglobin content were also evaluated. EAU treatment significantly (P<0.05) decreased tumor volume, packed cell volume and viable cell count, and increased the lifespan of EAC tumor-bearing mice. Hematological profiles were restored to normal levels in EAU-treated mice compared with EAC control mice. Taken together, these results suggest that the Aponogeton undulatus extract may have potent antioxidant and anticancer properties.

Antimicrobial peptide from Bacillus subtilis CSB138: characterization, killing kinetics, and synergistic potency.

We studied the prospect of synergy between the antimicrobial peptide p138c and non-peptide antibiotics for increasing the potency and bacterial killing kinetics of these agents. The production of p138c was maximized in the late exponential growth phase of Bacillus subtilis CSB138. Purification of p138c resulted in a total of 4800 arbitrary units (AU) with 19.15-fold and 3.2% recovery. Peptide p138c was thermo-tolerant up to 50 °C and stable at pH 5.8 to 11. The biochemical nature of p138c was determined by a bioassay, similar to tricine-SDS-PAGE, indicating inhibition at 3 kDa. The amino acid sequence of p138c was Gly-Leu-Glu-Glu-Thr-Val-Tyr-Ile-Tyr-Gly-Ala-Asn-Met-X-Ser. Potency and killing kinetics against vancomycin-resistant Staphylococcus aureus improved considerably when p138c was synergized with oxacillin, ampicillin, and penicillin G. The minimal inhibitory concentration (MIC) of p138c showed a 4-, 8-, and 16-fold improvement when p138c was combined with oxacillin, ampicillin, and penicillin G, respectively. The fractional inhibitory concentration index for the combination of p138c and oxacillin, ampicillin, and penicillin G was 0.3125, 0.25, and 0.09, respectively. Synergy with non-peptide antibiotics resulted in enhanced killing kinetics of p138c. Hence, the synergy between antimicrobial peptide and non-peptide antibiotics may enhance the potency and bacterial killing kinetics, providing more potent and rapidly acting agents for therapeutic use. [Int Microbiol 20(1):43-53 (2017)].

Identification of the biologically active constituents of Camellia japonica leaf and anti-hyperuricemic effect in vitro and in vivo.

Camellia japonica L. is a plant of which the seeds are used as a folk medicine, and it is native to South Korea, Japan and China. In previous study, triterpenes, flavonoids, tannins and fatty acids which have antiviral, antioxidant and anti inflammatory activity were reported from C. japonica leaf and flower. In Korea, the seed from this plant is used as a traditional medicine and in folk remedies for the treatment of bleeding and inflammation. However, the major issue associated with the use of the seed as a medicinal and/or functional food ingredient is its application to the pharmaceutical and food industry. First, the productivity of seed extract is very much less than that of the leaf. Second, the beneficial usage of the seed extract as an alternative medicine and functional source is not yet clear. Thus, in this study, we focused on another part of the plant, the leaf, and found that the extract of Camellia japonica leaf has a high concentration of vitamin E, rutin and other biologically active compounds related to hyperuricemia. We aimed to investigate the biological activities, namely the antioxidant activities, xanthine oxidase (XO) inhibitory activity and anti­hyperuricemic effects of extract from C. japonica leaf and the phytochemicals contained therein. Ethanol extracts of C. japonica leaf (ECJL) were prepared, and the extract was used with respect to antioxidant activities, total phenolic contents and XO inhibitory activity. The in vivo XO inhibitory activity and anti­hyperuricemic effects of the extract were evaluated in mice with potassium oxonate­induced hyperuricemia. To clarify the marker compounds that are responsible for the anti­hyperuricemic effects, several key constituents were identified using gas chromatography­mass spectrometry (GC­MS) and and liquid chromatography-mass spectrometry (LC-MS). ECJL was found to have strong antioxidant activities, and in vitro XO inhibitory activity. The results of the in vivo experiments using mice demonstrated that ECJL at the doses of 100 and 300 mg/kg inhibited hepatic XO activity and significantly attenuated hyperuricemia. To the best of our knowledge, the present study is the first report on the XO inhibitory and anti-hyperuricemic effects of ECJL, which can be therapeutically applied in the treatment of hyperuricemia and gout.

A novel multifunctional peptide oligomer of bacitracin with possible bioindustrial and therapeutic applications from a Korean food-source Bacillus strain.

CONCLUSION: CSP32 has stable characteristics and may find bio-industrial and therapeutic applications.

Antimicrobial and Anti-Inflammatory Effects of Ethanol Extract of Corylopsis coreana Uyeki Flos.

BACKGROUND: Corylopsis coreana Uyeki (Hamamelidaceae) is a medicinal plant cultivated in Northeast Asia. Previously, we have reported that an ethanol extract of Corylopsis coreana Uyeki flos (ECCF) contains four active compounds with antioxidant activity. OBJECTIVE: The aim of this study was to investigate the antimicrobial spectrum against infectious bacteria and anti-inflammatory effect of ECCF in a mouse model of acute local inflammation. MATERIALS AND METHODS: In vitro antimicrobial susceptibility was evaluated using standard plate assay technique. Antimicrobial activities (minimum inhibitory concentration, MIC; µg/mL) were determined with the serial dilution method. In vivo anti-inflammatory activity was studied using a mouse model of carrageenan-induced air pouch inflammation. RESULTS: The ECCF showed antimicrobial activities against general bacteria and drug-resistant bacteria including Staphylococcus aureus, Micrococcus luteus ATCC 9341, Mycrobacterium smegmatis ATCC 9341, Mycrobacterium smegmatis ATCC 9341, Salmonella typhimrium KCTC 1925, and nine methicillin-resistant Staphylococcus aureus strains, with MIC values ranging from 250 to 1000 µg/mL. In in vivo mouse model, inflammatory morphologic changes observed in the air pouch membrane were restored to its normal condition by the ECCF treatment. Moreover, the ECCF significantly reduced exudate volumes, protein contents, inflammatory cell counts, and pro-inflammatory cytokine levels in the exudates recovered from air pouches of the mouse model. Flavonoids in the ECCF were found to contain bergenin, quercitrin, and quercetin with reported anti-inflammatory activity via suppressing tumor necrosis factor-α production. CONCLUSION: To the best of our knowledge, this is the first report to demonstrate the antimicrobial and anti-inflammatory activities of ECCF. Our results suggest that the ECCF might potentially serve as an alternative or complementary medicine for treating inflammatory diseases caused by microbial infection. SUMMARY: ECCF showed antimicrobial activity against infectious bacteria and multidrug-resistant strains.ECCF exhibited anti-inflammatory activity in a carrageenan-induced air pouch mouse model.ECCF contained several active constituents such as bergenin, quercitrin, and quercetin. Abbreviations used:Corylopsis coreana CCF: Uyeki flos, ECCF: ethanol extract of CCF.

A multifunctional alanine-rich anti-inflammatory peptide BCP61 showed potent inhibitory effects by inhibiting both NF-κB and MAPK expression.

The purified BCP61 was reported to be a unique low-molecular-weight (MW) anti-microbial peptide because of its non-identical alanine-rich N-terminal sequence. In this study, we investigated the anti-inflammatory effects of BCP61 on induction of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), pro-inflammatory cytokines, nuclear factor-kappa B (NF-κB), and mitogen-activated protein kinases (MAPKs) in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. The treatment with BCP61, with varying concentrations of 10, 50, and 100 µg/mL, inhibited levels of expression of LPS-induced NF-κB and MAPKs (extracellular signal-related kinases (ERKs), c-Jun NH2-terminal kinase (JNK), and mitogen-activated protein (p38)) as well as production of pro-inflammatory mediators, such as nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6). The results suggested that BCP61 prevents inhibitor of kappa B (IκBα) phosphorylation and degradation, thereby inhibiting the nuclear translocation of the p65 protein. We do report that the use of BCP61 in the treatment of inflammation as well as microbial infection could be a potent therapeutic candidate.