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The need to discover new types of antimicrobial agents has grown since the emergence of antibiotic-resistant pathogens that threaten human health. The world's oceans, comprising complex niches of biodiversity, are a promising environment from which to extract new antibiotics-like compounds. In this study, we newly isolated

Antibiotics have been used in various fields such as livestock farm and fish farm as well as hospital in order to treat diseases caused by bacteria. However, the antibiotics that are not completely decomposed, but remains as residue and discharge to aquatic environment, can cause an imbalance in the gut flora of host, as well as regulate abnormal host gene regulatory system. We investigated the effects of chronic exposure with the low concentrations of erythromycin and ampicillin on gut microbiome and immune and stress-related gene expression using Korea native ricefish (Oryzias latipes). As a result of microbiome analysis, the proportion of Proteobacteria was increased in the ricefish when exposed to erythromycin and ampicillin chronically, whereas the proportion of other bacterial phyla decreased. In addition, the immune and stress-related genes were significantly influenced in the ricefish under the chronic antibiotics exposure. These results show that the internal microbial flora and the host gene expression are susceptible even in the low concentration of chronic antibiotic existing environments. This study provides the importance of the appropriate use of antibiotics dose to maintain the sustainable and healthy aquaculture industry and water ecosystem.

No effective cryopreservation technique exists for fish eggs and embryos; thus, the cryopreservation of germ cells (spermatogonia or oogonia) and subsequent generation of eggs and sperm would be an alternative solution for the long-term preservation of piscine genetic resources. Nevertheless, in our previous study using rainbow trout, we showed that recipients transplanted with XY spermatogonia or XX oogonia produced unnatural sex-biased F1 offspring. To overcome these obstacles, we transplanted immature germ cells (XX oogonia or XY spermatogonia; frozen for 33 days) into the body cavities of triploid hatchlings, and the transplanted germ cells possessed a high capacity for differentiating into eggs and sperm in the ovaries and testes of recipients. Approximately 30% of triploid recipients receiving frozen germ cells generated normal salmon that displayed the donor-derived black body color phenotype, although all triploid salmon not receiving transplants were functionally sterile. Furthermore, F1 offspring obtained from insemination of the oogonia-derived eggs and spermatogonia-derived sperm show a normal sex ratio of 1:1 (female:male). Thus, this method presented a critical technique for practical conservation projects for other teleost fish species and masu salmon.

The goal of this study was to evaluate the influence of various supplementary feeds on the chemical composition and production of bioactive substances in Protaetia brevitarsis larvae. The primary feed-oak-fermented sawdust-was supplemented with a variety of substances, including aloe, apple, banana, sweet persimmon (S. persimmon) and sweet pumpkin (S. pumpkin). Crude protein and fat content were the highest in the control and S. pumpkin group, respectively. Supplementary feeds increased the content of unsaturated fatty acids, except in the group receiving S. pumpkin, in which oleic acid was the most abundant (58.2%-64.5%). Free essential amino acids in larvae receiving supplementary aloe were higher compared with the control group except for Lys and His. Polyphenol and flavonoid contents and the antioxidant activities of ABTS and DPPH were higher in all treated groups compared with the control group. Although supplementary feeds led to a decreased crude protein content in the treated larvae when compared with the control group, these treatments generally improved the levels of unsaturated fatty acids and antioxidative activity. Therefore, we suggest that among the supplementary foods tested, aloe is a better resource for P. brevitarsis based on crude protein content, free amino acids and other bioactive compounds such as unsaturated fatty acids and antioxidants.

In this study, genetic engineering methods, ranging from gene transformation to gene editing, were efficiently conducted on cotyledon explants in Solanum nigrum. Organogenic calli were observed on the explants following 10 days of growth on medium supplemented with 2 mg/L zeatin and 0.2 mg/L indole acetic acid (IAA), which we suggest to be an efficient shoot initiation medium (SM1). In vitro shoot production was enhanced in explants grown on media supplemented with 1 mg/L zeatin and 0.1 mg/L IAA, which we used as a proper shoot differentiation medium (SM2) in S. nigrum shoot regeneration. Direct infection of explants with Agrobacterium harboring CRISPR/Cas9 constructs can simplify the method of Agrobacterium-mediated T-DNA transformation by skipping the preincubation step. The method was applied to deliver transgenes for genome editing, such as CRISPR/Cas9 DNA. SnLazy1 locus, considered to be orthologs of tomato Lazy1, was edited using the CRISPR/Cas9 system in S. nigrum. Two independent deletion snlazy1-cr alleles were successively inherited and showed stem growth in a relatively downward direction. Our method offers rapid and simple procedure for gene transformation and genome editing that could be applicable for the enhancement of beneficial traits for precision plant breeding and engineering quantitative trait loci in S. nigrum.

The claim that organic agriculture produces higher levels of phytochemicals has been controversial for decades. Using strawberries as a model crop in field conditions, a preharvest leaf wounding stress was applied to study the production of phytochemicals in fruits. As a result phenolic compounds (PCs) and total soluble sugars increased significantly, where specific phenylpropanoids showed increment up to 137% and several genes related to PCs biosynthesis and sugar transport were overexpressed. It was observed that the accumulation of PCs on fruits can be triggered by the application of wounding stress in a distant tissue and this accumulation is directly related to carbon partition and associated gene expression. This supports the idea that higher levels of healthy phytochemicals reported in organic fruits and vegetables could be due to the wounding component of the biotic stress attributed to insects to which the plant are exposed to.

Solanum nigrum, known as black nightshade, is a medicinal plant that contains many beneficial metabolites in its fruit. The molecular mechanisms underlying the synthesis of these metabolites remain uninvestigated due to limited genetic information. Here, we identified 47,470 unigenes of S. nigrum from three different tissues by de novo transcriptome assembly, and 78.4% of these genes were functionally annotated. Moreover, gene ontology (GO) analysis using 18,860 differentially expressed genes (DEGs) revealed tissue-specific gene expression regulation. We compared gene expression patterns between S. nigrum and tomato (S. lycopersicum) in three tissue types. The expression patterns of carotenoid biosynthetic genes were different between the two species. Comparison of the expression patterns of flavonoid biosynthetic genes showed that 9 out of 14 enzyme-coding genes were highly upregulated in the fruit of S. nigrum. Using CRISPR-Cas9-mediated gene editing, we knocked out the R2R3-MYB transcription factor SnAN2 gene, an ortholog of S. lycopersicum ANTHOCYANIN 2. The mutants showed yellow/green fruits, suggesting that SnAN2 plays a major role in anthocyanin synthesis in S. nigrum. This study revealed the connection between gene expression regulation and corresponding phenotypic differences through comparative analysis between two closely related species and provided genetic resources for S. nigrum.

This study characterized the antimicrobial substances produced by the radiation-resistant bacterium Nibribacter radioresistens. The antimicrobial substances showed activity against Salmonella Gallinarum, pathogenic Escherichia coli, Bacillus cereus, Streptococcus iniae, and Saccharomyces cerevisiae. The substances showed higher activity against Gram-positive bacteria than against Gram-negative bacteria and yeast. N. radioresistens showed the best growth rate in LB liquid medium at 37oC; however, production of the antimicrobial substances was not associated with growth. Since the activity of the antimicrobial substances was affected by proteinase K and EDTA, the substances were presumed to be antimicrobial peptides (AMPs). The antimicrobial substances produced by N. radioresistens were unstable at higher temperatures and in acidic and basic pH ranges, and most of the activity was attributed to either low ( 30 kDa) molecules. When S. Gallinarum was treated with the antimicrobial substances, the cell destruction was acted on the cell envelope. Therefore, we concluded that N. radioresistens produces broad-spectrum and very unstable antimicrobial substances that mostly consist of low- and high-molecular weight peptides.

Self-assembly of peptides containing both l- and d-isomers often results in nanostructures with enhanced properties compared to their enantiomeric analogues, such as faster kinetics of formation, higher mechanical strength, and enzymatic stability. However, occurrence and consequences of the heterochiral assembly in the cellular microenvironment are unknown. In this study, we monitored heterochiral assembly of amphiphilic peptides inside the cell, specifically mitochondria of cancer cells, resulting in nanostructures with refined morphological and biological properties owing to the superior interaction between the backbones of opposite chirality. We have designed a mitochondria penetrating tripeptide containing a diphenyl alanine building unit, named as Mito-FF due to their mitochondria targeting ability. The short peptide amphiphile, Mito-FF co-assembled with its mirror pair, Mito-ff, induced superfibrils of around 100 nm in diameter and 0.5-1 μm in length, while enantiomers formed only narrow fibers of 10 nm in diameter. The co-administration of Mito-FF and Mito-ff in the cell induced drastic mitochondrial disruption both

BACKGROUND/OBJECTIVES Vascular inflammation is an important feature in the atherosclerotic process. Recent studies report that leaves and branches of Carpinus turczaninowii (C. turczaninowii) have antioxidant capacity and exert anti-inflammatory effects. However, no study has reported the regulatory effect of C. turczaninowii extract on the arterial inflammatory response. This study therefore investigated modulation of the arterial inflammatory response after exposure to C. turczaninowii extract, using human aortic vascular smooth muscle cells (HAoSMCs). MATERIALS/METHODS Scavenging activity of free radicals, total phenolic content (TPC), cell viability, mRNA expressions, and secreted levels of cytokines were measured in LPS-stimulated (10 ng/mL) HAoSMCs treated with the C. turczaninowii extract. RESULTS C. turczaninowii extract contains high amounts of TPC (225.6 ± 21.0 mg of gallic acid equivalents/g of the extract), as well as exerts time-and dose-dependent increases in strongly scavenged free radicals (average 14.8 ± 1.97 µg/mL IC50 at 40 min). Cell viabilities after exposure to the extracts (1 and 10 µg/mL) were similar to the viability of non-treated cells. Cytokine mRNA expressions were significantly suppressed by the extracts (1 and 10 µg/mL) at 6 hours (h) after exposure. Interleukin-6 secretion was dose-dependently suppressed 2 h after incubation with the extract, at 1-10 µg/mL in non-stimulated cells, and at 5 and 10 µg/mL in LPS-stimulated cells. Similar patterns were also observed at 24 h after incubation with the extract (at 1-10 µg/mL in non-stimulated cells, and at 10 µg/mL in the LPS-stimulated cells). Soluble intracellular vascular adhesion molecules (sICAM-1) secreted from non-stimulated cells and LPS-stimulated cells were similarly suppressed in a dose-dependent manner after 24 h exposure to the extracts, but not after 2 h. In addition, sICAM-1 concentration after 24 h treatment was positively related to IL-6 levels after 2 h and 24 h exposure (r = 0.418, P = 0.003, and r = 0.524, P < 0.001, respectively). CONCLUSIONS This study demonstrates that C. turczaninowii modulates the arterial inflammatory response, and indicates the potential to be applied as a therapeutic use for atherosclerosis.

Antibiotics have been widely used to inhibit microbial growth and to control bacterial infection; however, they can trigger an imbalance in the gut flora of the host and dysregulate the host gene regulatory system when discharged into the aquatic environment. We investigated the effects of chronic exposure to a low concentration of erythromycin and ampicillin, focusing on gut microbiome and global gene expression profiles from Korea native ricefish (Oryzias latipes). The proportion of Proteobacteria (especially the opportunistic pathogen Aeromonas veronii) was significantly increased in the ricefish under the chronic exposure to erythromycin and ampicillin, whereas that of other bacterial phyla (i.e., Fusobacteria) decreased. In addition, the expression of genes involved in immune responses such as chemokines and immunocyte chemotaxis was significantly influenced in ricefish in the aquatic environment with antibiotics present. These results show that the internal microbial flora and the host gene expression are susceptible even at a low concentration of chronic antibiotics in the environment, supporting the importance of the appropriate use of antibiotic dose to maintain the sustainable and healthy aquaculture industry and water ecosystem.

The recent pandemic of coronavirus disease 2019 (COVID-19) has increased demand for chemical disinfectants, which can be potentially hazardous to users. Here, we suggest that the cell-free supernatant from Lactobacillus plantarum NIBR97, including novel bacteriocins, has potential as a natural alternative to chemical disinfectants. It exhibits significant antibacterial activities against a broad range of pathogens, and was observed by scanning electron microscopy (SEM) to cause cellular lysis through pore formation in bacterial membranes, implying that its antibacterial activity may be mediated by peptides or proteins and supported by proteinase K treatment. It also showed significant antiviral activities against HIV-based lentivirus and influenza A/H3N2, causing lentiviral lysis through envelope collapse. Furthermore, whole-genome sequencing revealed that NIBR97 has diverse antimicrobial peptides, and among them are five novel bacteriocins, designated as plantaricin 1 to 5. Plantaricin 3 and 5 in particular showed both antibacterial and antiviral activities. SEM revealed that plantaricin 3 causes direct damage to both bacterial membranes and viral envelopes, while plantaricin 5 damaged only bacterial membranes, implying different antiviral mechanisms. Our data suggest that the cell-free supernatant from L. plantarum NIBR97, including novel bacteriocins, is potentially useful as a natural alternative to chemical disinfectants.

Phenol and formalin are major water pollutants that are frequently discharged into the aquatic milieu. These chemicals can affect broad domains of life, including microorganisms. Aquatic pollutants, unlike terrestrial pollutants, are easily diluted in water environments and exist at a sub-inhibitory concentration (sub-IC), thus not directly inhibiting bacterial growth. However, they can modulate gene expression profiles. The sub-IC values of phenol and formalin were measured by minimal inhibitory concentration (MIC) assay to be 0.146% (1.3 mM) and 0.0039% (0.38 mM), respectively, in Edwardsiella piscicida CK108, a Gram-negative fish pathogen. We investigated the differentially expressed genes (DEG) by RNA-seq when the cells were exposed to the sub-ICs of phenol and formalin. DEG analyses revealed that genes involved in major virulence factors (type I fimbriae, flagella, type III and type VI secretion system) and various cellular pathways (energy production, amino acid synthesis, carbohydrate metabolism and two-component regulatory systems) were up- or downregulated by both chemicals. The genome-wide gene expression data corresponded to the results of a quantitative reverse complementary-PCR and motility assay. This study not only provides insight into how a representative fish pathogen, E. piscicida CK108, responds to the sub-ICs of phenol and formalin but also shows the importance of controlling chemical pollutants in aquatic environments.

The prevalence of antibiotic-resistant bacteria has become an immediate threat to public health. Antimicrobial peptides are attracting attention as a new source of antibiotics due to their ability to prevent drug-resistances with fewer side effects. Spider venom is composed of various bioactive substances with multiple functionalities such as antimicrobial and anti-inflammatory effects. Here, RNA sequencing was conducted on the venom gland of the spider Pardosa astrigera, and a potential toxin peptide with antibacterial properties was selected via homology and in silico analysis. A novel toxin, Lycotoxin-Pa4a, inhibited both gram-negative and gram-positive bacteria by disrupting the outer and bacterial cytoplasmic membrane. Moreover, the peptide downregulated the expression of pro-inflammatory mediators while upregulating the level of anti-inflammatory cytokine by inactivating mitogen-activated protein kinase signaling in a lipopolysaccharide-stimulated murine macrophage cell line. In this research, we identified a novel peptide toxin, Lycotoxin-pa4a, with antibacterial and anti-inflammatory properties, suggesting its potential for the development of a new antibiotics, as well as offering insights into the utilization of biological resources.

Phenol and formalin are major water pollutants that are frequently discharged into the aquatic milieu. These chemicals can affect broad domains of life, including microorganisms. Aquatic pollutants, unlike terrestrial pollutants, are easily diluted in water environments and exist at a sub-inhibitory concentration (sub-IC), thus not directly inhibiting bacterial growth. However, they can modulate gene expression profiles. The sub-IC values of phenol and formalin were measured by minimal inhibitory concentration (MIC) assay to be 0.146% (1.3 mM) and 0.0039% (0.38 mM), respectively, in Edwardsiella piscicida CK108, a Gram-negative fish pathogen. We investigated the differentially expressed genes (DEG) by RNA-seq when the cells were exposed to the sub-ICs of phenol and formalin. DEG analyses revealed that genes involved in major virulence factors (type I fimbriae, flagella, type III and type VI secretion system) and various cellular pathways (energy production, amino acid synthesis, carbohydrate metabolism and two-component regulatory systems) were up- or downregulated by both chemicals. The genome-wide gene expression data corresponded to the results of a quantitative reverse complementary-PCR and motility assay. This study not only provides insight into how a representative fish pathogen, E. piscicida CK108, responds to the sub-ICs of phenol and formalin but also shows the importance of controlling chemical pollutants in aquatic environments.

Edwardsiella piscicida CK41 is a fish-pathogenic Gram-negative bacterium isolated from diseased flounder in the Republic of Korea. Here, we report the genome sequence of E. piscicida CK41, comprising one chromosome of 3.76 Mbp and one plasmid of 72.7 kbp. A total of 3,406 protein-coding genes, 98 tRNAs, and 25 rRNAs are predicted to be present in the genome.

Ellagic acid (EA) is a component of ellagitannins, present in crops such as pecans, walnuts, and many berries, which metabolized by the gut microbiota forms urolithins A, B, C, or D. In this study, ellagic acid, as well as urolithins A and B, were tested on 3T3-L1 preadipocytes for differentiation and lipid accumulation. In addition, inflammation was studied in mature adipocytes challenged with lipopolysaccharide (LPS). Results indicated that EA and urolithins A and B did not affect differentiation (adipogenesis) and only EA and urolithin A attenuated lipid accumulation (lipogenesis), which seemed to be through gene regulation of glucose transporter type 4 (GLUT4) and adiponectin. On the other hand, gene expression of cytokines and proteins associated with the inflammation process indicate that urolithins and EA differentially inhibit tumor necrosis factor alpha (TNF&alpha, ), inducible nitric oxide synthase (iNOS), interleukin 6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1). Urolithins A and B were found to reduce nuclear levels of phosphorylated nuclear factor &kappa, B (p-NF-&kappa, B), whereas all treatments showed expression of nuclear phosphorylated protein kinase B (p-AKT) in challenged LPS cells when treated with insulin, indicating the fact that adipocytes remained insulin sensitive. In general, urolithin A is a compound able to reduce lipid accumulation, without affecting the protein expression of peroxisome proliferator-activated receptor-&gamma, (PPAR&gamma, ), CCAAT/enhancer binding protein-&alpha, (c/EBP&alpha, ), and PPAR&alpha, whereas EA and urolithin B were found to enhance PPAR&gamma, and c/EBP&alpha, protein expressions as well as fatty acid (FA) oxidation, and differentially affected lipid accumulation.

The goal of this study was to evaluate the influence of various supplementary feeds on the chemical composition and production of bioactive substances in Protaetia brevitarsis larvae. The primary feed—oak-fermented sawdust—was supplemented with a variety of substances, including aloe, apple, banana, sweet persimmon (S. persimmon) and sweet pumpkin (S. pumpkin). Crude protein and fat content were the highest in the control and S. pumpkin group, respectively. Supplementary feeds increased the content of unsaturated fatty acids, except in the group receiving S. pumpkin, in which oleic acid was the most abundant (58.2%–64.5%). Free essential amino acids in larvae receiving supplementary aloe were higher compared with the control group except for Lys and His. Polyphenol and flavonoid contents and the antioxidant activities of ABTS and DPPH were higher in all treated groups compared with the control group. Although supplementary feeds led to a decreased crude protein content in the treated larvae when compared with the control group, these treatments generally improved the levels of unsaturated fatty acids and antioxidative activity. Therefore, we suggest that among the supplementary foods tested, aloe is a better resource for P. brevitarsis based on crude protein content, free amino acids and other bioactive compounds such as unsaturated fatty acids and antioxidants.

To identify and compare venom components and expression patterns, venom gland-specific transcriptome analyses were conducted for 14 Aculeate bees and wasps. TPM (transcripts per kilobase million) values were normalized using the average transcription level of a reference housekeeping gene (dimethyladenosine transferase). Orthologous venom component genes across the 14 bee and wasp species were identified, and their relative abundance in each species was determined by comparing normalized TPM values. Based on signal sequences in the transcripts, the genes of novel venom components were identified and characterized to encode potential allergens. Most of the allergens and pain-producing factors (arginine kinase, hyaluronidase, mastoparan, phospholipase A1, phospholipase A2, and venom allergen 5) showed extremely high expression levels in social wasps. Acid phosphatase, neprilysin, and tachykinin, which are known allergens and neurotoxic peptides, were found in the venom glands of solitary wasps more often than in social wasps. In the venom glands of bumblebees, few or no transcripts of major allergens or pain-producing factors were identified. Taken together, these results indicate that differential expression patterns of the venom genes in some Aculeate species imply that some wasps and bumblebee species have unique groups of highly expressed venom components. Some venom components reflected the Aculeate species phylogeny, but others did not. This unique evolution of specific venom components in different groups of some wasps and bumblebee species might have been shaped in response to both ecological and behavioral influences.

Endophytes, important plant-associated mycobionts, have attracted a great deal of attention because of their bioactive secondary metabolites. Even though halophytes have been reported to overcome salt stress via associations with their endophytes, few studies have investigated the metabolites produced by the endophytes from halophytes. In this study, a dark septate endophytic fungal strain (JS0464), identified as Gaeumannomyces sp. by ITS sequencing, was isolated from the rhizome of a halophyte, Phragmites communis, in Suncheon bay, South Korea. This strain was cultured on a large scale and extracted with ethyl acetate. Chemical investigations of extracts of JS0464 led to the isolation of two glycosylated dialkylresorcinol derivatives (1-2), an anthraquinone derivative (3) and eight known compounds (4-11), which were identified by spectroscopic analyses incorporating one-dimensional/2D NMR and MS. Nine compounds showed significant nitric oxide reduction activity in lipopolysaccharide-stimulated microglia BV-2 cells, seven of which did not impair cell viability. The results suggest that endophytes from the halophytes could be potential resources for bioactive natural products.

This study was done to analyze the antimicrobial activity of gut-derived microbes of Protaetiabrevitarsisseulensis larvae. Among the lactic acid bacterial strains isolated from the intestinal microbes of P. brevitarsisseulensis larvae, 10 strains were firstly selected from results of antimicrobial activity against Salmonella Typhimurium χ3339 strain as a susceptible strain. As a result, since the Minimum Inhibitory Concentration (MIC) was the best result in the K9 strain, further tests were performed on the K9 strain in subsequent studies. The K9 strain was identified as Lactobacillus plantarum from 16 SrDNA sequencing, named as L. plantarum K9. The antibacterial activity according to the amount of susceptible strain showed no or lower activity at 108 and 107 CFU/ml, whereas stronger antibacterial activity below 106 CFU/ml. The antibacterial activity test according to the cultural temperature showed the strongest antibacterial activity at 37℃. In the thermal stability test, it was confirmed that the antimicrobial activity was maintained up to 50℃ but significantly decreased from 60℃ or higher. However, some of the AMPs exhibited thermal stability because of remaining the activity above 60℃ or higher. Since proteinase K treatmentdecreases the antibacterial activity, we confirm that antibacterial activity is done by peptide-based bioactive substances. In summary, we suggest that AMPs in L. plantarum K9 strain are a potential of application as a natural antibacterial agent owing to thermal stability.

As an attempt to establish a method for efficient and safe administration of therapeutic carbon monoxide (CO) to the human body, supramolecular nanoplatforms incorporated with CO-releasing molecules (CORMs) have recently been developed. In particular, hydrogel scaffolds have attracted considerable attention due to the possibility of site-specific and controlled liberation of CO. However, it would be greatly beneficial to enhance the mechanical strength of hydrogels to widen their applicability in biomedical, pharmaceutical, and surgical sectors. Herein, we report a visible light-mediated crosslinkable supramolecular CO-releasing hydrogel (CORH), based on the fibrillar assembly of elastomeric protein-derived tyrosine-containing short peptides. A photo-driven dimerization of tyrosine moieties located on the fibrillar surface of CORH, accelerated by a Ru-based catalyst, results in the entanglement and bundling of nanofibrils that significantly increases the mechanical strength and stability of the CORH, which allows prolonged CO-liberation through limiting the contact of CORMs with water molecules. The contact probability of a CORM with water determined by the spatial position of the CORM on the fibrils containing a crosslinkable tyrosine moiety that affects CO-releasing behavior was confirmed by adjusting the CORM position closer to or farther from the tyrosine in the peptide sequence. A bulky CORM closely located to the tyrosine in a peptide inhibited the effective dityrosine formation of tyrosine on the fibril surface, resulting in loose bundling of nanofibrils in the CORH and facilitating the release of CO through the exchange with water. The photo-crosslinked CORH demonstrated a potent cytoprotective effect on oxidatively stressed cardiomyocytes, as expected. This work could provide a useful insight for the practical application of gasotransmitters as functional nanomaterials in pharmaceutical and biomedical fields.

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Exposure to particulate matter (PM) in ambient air is known to increase the risk of cardiovascular disorders and mortality. The cytotoxicity of PM is mainly due to the abnormal increase of reactive oxygen species (ROS), which damage cellular components such as DNA, RNA, and proteins. The correlation between PM exposure and human disorders, including mortality, is based on long-term exposure. In this study we have investigated acute responses of mucus-secreting goblet cells upon exposure to PM derived from a heavy diesel engine. To this end, we employed the mucociliary epithelium of amphibian embryos and human Calu-3 cells to examine PM mucotoxicity. Our data suggest that acute exposure to PM significantly impairs mucus secretion and results in the accumulation of mucus vesicles in the cytoplasm of goblet cells. RNA-seq analysis revealed that acute responses to PM exposure significantly altered gene expression patterns; however, known regulators of mucus production and the secretory pathway were not significantly altered. Interestingly, pretreatment with α-tocopherol nearly recovered the hyposecretion of mucus from both amphibian and human goblet cells. We believe this study demonstrates the mucotoxicity of PM and the protective function of α-tocopherol on mucotoxicity caused by acute PM exposure from heavy diesel engines.

Hyperglycemia-induced oxidative stress triggers severe vascular damage and induces an inflammatory vascular state, and is, therefore, one of the main causes of atherosclerosis. Recently, interest in the natural compound Carpinus turczaninowii has increased because of its reported antioxidant and anti-inflammatory properties. We investigated whether a C. turczaninowii extract was capable of attenuating high glucose-induced inflammation and arterial damage using human aortic vascular smooth muscle cells (hASMCs). mRNA expression levels of proinflammatory response [interleukin-6 (IL-6), tumor necrosis factor-&alpha, (TNF-&alpha, )], endoplasmic reticulum (ER) stress [CCAAT-enhancer-binding proteins (C/EBP) homologous protein (CHOP)], and adenosine monophosphate (AMP)-protein activated kinase &alpha, 2 (AMPK &alpha, 2)], and DNA damage [phosphorylated H2.AX (p-H2.AX)] were measured in hASMCs treated with the C. turczaninowii extracts (1 and 10 &mu, g/mL) after being stimulated by high glucose (25 mM) or not. The C. turczaninowii extract attenuated the increased mRNA expression of IL-6, TNF-&alpha, and CHOP in hASMCs under high glucose conditions. The expression levels of p-H2.AX and AMPK &alpha, 2 induced by high glucose were also significantly decreased in response to treatment with the C. turczaninowii extract. In addition, 15 types of phenolic compounds including quercetin, myricitrin, and ellagic acid, which exhibit antioxidant and anti-inflammatory properties, were identified in the C. turczaninowii extract through ultra-performance liquid chromatography-quadrupole-time of flight (UPLC-Q-TOF) mass spectrometry. In conclusion, C. turczaninowii may alleviate high glucose-induced inflammation and arterial damage in hASMCs, and may have potential in the treatment of hyperglycemia-induced atherosclerosis.

We identified vespid chemotactic peptide (VCP) and vespakinin (Vespk) from the lesser paper wasp, Parapolybia varia . The cDNA, genomic DNA, and mature peptide sequences of P. varia VCP ( Pv VCP) and Vespk ( Pv Vespk) were determined. To investigate the pharmacological and toxicological properties of Pv VCP and Pv Vespk, their hemolytic, anti-microbial, anti-fungal, and anti-tumor activities were evaluated and compared with those of Vespa mandarina VCP ( Vm VCP) and Vespk ( Vm Vespk). Pv VCP, Pv Vespk, and Vm Vespk showed little to low hemolytic activities. Only Vm VCP showed hemolytic activity at a high concentration. Among the four peptides tested, Vm VCP showed both anti-microbial and anti-fungal activities, whereas Pv VCP showed only anti-fungal activity to Candida albicans . Interestingly, Pv VCP showed significantly stronger anti-tumor activities to two ovarian cancer cell lines compared with Vm VCP. Vespks only showed anti-tumor activity to SK-OV-3 cells but not to NIH-OVCAR-3 cells. These differences in anti-tumor activity might have been caused by the differences in secondary structure among peptides. A circular dichroism spectrometry analysis revealed that VCPs have more amphiphilic α-helix structures than Vespks. Taken together, the low hemolytic but strong anti-tumor activities of Pv VCP suggest that this peptide could be a candidate for developing a new anti-tumor peptide drug or drug carrier in the future.

Multifunctional nanomedicines with imaging and multimodal therapies have become a new trend in the current development of cancer therapy. Herein, we report the use of a Flash NanoPrecipitation approach for fabricating a multifunctional nanoparticulate theranostic system (MNTS) simultaneously encapsulating paclitaxel, gold nanoparticles, and iron oxide nanoparticles in poly(ethylene oxide)-block-poly(e-caprolactone) (PEO-b-PCL). The combination of these building blocks in a single system translates into an advance nanomedicine with concurrent chemotherapeutic, photothermal therapeutic, and MRI contrast properties. Using dynamic light scattering, the hydrodynamic diameter of MNTS was found to range from 50 nm to 500 nm with the mean size of 190 ± 14 nm. The co-localization of paclitaxel, gold nanoparticles, and iron oxide nanoparticles were confirmed via scanning- and cryo-transmission electron microscopy. At a paclitaxel concentration of 100 pM, the cell viability after a 72 h treatment of MNTS was 35 ± 3% and 31 ± 1% for MCF-7 and MDA-MB-231 cell lines, respectively while that of bare paclitaxel was 77 ± 2% and 91 ± 3%, respectively. Irradiation of MNTS with a 200 mW laser (532 nm) for 5 min during the treatment stage resulted in a further 20% and 226% decrease in the viability of MCF-7 and MDA-MB-231, respectively. MRI relaxivity measurements revealed that the T2 relaxation times of MNTS was 47 ± 5 ms and significantly different from that of most human anatomical parts susceptible to cancer.

Despite the versatile metabolic functions of peroxisomes such as lipid synthesis and fatty acid oxidation and their relevance to genetically inherited diseases, namely, peroxisome biogenesis disorders and peroxisomal enzyme deficiency, there is not much research on peroxisome-targeting therapeutics. Herein we present supramolecular nanostructured probes based on the self-assembly of peptide amphiphiles (PAs) having peroxisome-targeting ability in mammalian cells. The PA was designed to include the peroxisome-targeting tripeptide (SKL) and a fluorescent dye (pyrene). It was revealed that the presence of the SKL-appended carboxyl terminal group of PA, the extent of α-helical nature of the peptide block, and the fibrillar morphology of nano-assemblies affected the targeting efficiency of PA supramolecular nanoprobe. The simple modification of PAs by the peroxisome-targeting strength prediction showed an enhanced peroxisome specificity, as expected. This work provides important insights into designing subcellular organelle-targeting nanoparticles for next-generation nanomedicines.

The Goal of this study was to identify whether the extract of Liriope platyphylla tuber can induce antiencephalitis. The yield of the hot water extraction was 1.5 times higher than the extract by simple homogenization. When the methanol fractionation after the hot water extract was done, the precipitated pellet in 70% methanol showed the highest antiencephalitis activity. In organic solvent partition after 70% methanol extract, the water phases in hexane, chloroform and ethyl acetate were exhibited higher activities, whereas its activity in butanol partition was divided into water and butanol layers. The eluents from the Sep-Pak R18 cartridge by fraction derived from the butanol and water phases in butanol partition showed the highest activity in the fraction of the hexane: ethyl acetate (5: 5) and ethyl acetate, respectively. It is assumed that the antiencephalitic activity is maintained not only in the hydrophilic materials but also in the hydrophobic ones. Therefore, we suggest that specific metabolites retained in L. platyphylla tuber are a high possibility to be applied as antiencephalitic agents.

Gadolinium (Gd[III])-based nanoaggregates are potential noninvasive magnetic resonance imaging (MRI) probes with excellent spatial and temporal resolution for cancer diagnosis. Peptides conjugated with Gd3+ can aid in supramolecular scaffolding for MRI nanoagents because of their inherent biocompatibility and degradability. We report here a strategy to tune the MR relaxivity of tumor cell-targeted nanoagents and enhance the antimicrobial and anticancer activities of nanoagents based on rationally designed antimicrobial peptide (AMP) assembly. A tripeptide with glycyl-l-histidyl-l-lysine (GHK) capable of Gd3+ chelation was attached to short AMPs containing pyrazole amino acids that spontaneously assembled as a function of the number of hydrophobic amino acid residues and the peptide length of AMPs. Aqueous coassembly of GHK with tumor-targeting, cyclic arginine-glycine-aspartic acid (cRGD)-tagged AMPs resulted in the formation of micelles, fibrils, vesicles, sheets, and planar networks. Interestingly, the ...

Mastoparans from the venom of social wasps have attracted considerable attention as effective antibiotic candidates. In this study, mastoparan V1 (MP-V1) from Vespula vulgaris was first disclosed to have a peptide amino acid sequence distinct from typical mastoparans and its biochemical properties and antimicrobial effects were compared with those of typical mastoparans MP-L, -X(V) and -B. Circular dichroism (CD) spectroscopy revealed that MP-V1 and -X(V) form more stable α-helical conformations in lipid membrane-like environments than MP-L and -B. In parallel, these two also showed more effective antimicrobial activities against the pathogens than did MP-L and -B. Although MP-V1 had a less stable α-helical conformation than MP-X(V), it showed stronger antimicrobial effects against Streptococcus mutans and Salmonella enterica than MP-X(V). In the meantime, analysis of hemolytic activity revealed a range of doses (~50 μM) that exhibited little potent cytotoxicity on human erythrocytes. Finally, the atypical MP-V1 peptide amino acid sequence provided important clues to understanding its antimicrobial mechanism from a structural perspective. Therefore, it has been concluded that MP-V1 is a de novo type of mastoparan with superior antimicrobial activities against both pathogenic bacteria and fungi, which may be useful in developing multipurpose antimicrobial drugs against infectious diseases.

Seventeen polyphenols (e.g, apigenin, genistein, and luteolin glycosides) and 11 lipophilic compounds (e.g., fatty acids, sterols, and terpenes) were detected by LC-MS/MS-ESI and GC-MS, respectively, in Jatropha platyphylla. Extracts from pulp, kernel, and leaves and fractions were studied to know their effect on some pro-inflammatory mediators. Phenolic and lipophilic extracts showed significant inhibitory effects on ROS and NO production while not affecting mitochondrial activity or superoxide generation rate in lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. In addition, NO production was also diminished by lipophilic leaf fractions F1 and F2 with the latter fraction showing a greater effect and composed mainly of sterols and terpene. Furthermore, total extracts showed nonselective inhibitions against cyclooxygenase COX-1 and COX-2 activities. All together, these results suggest that J. platyphylla extracts have potential in treating inflammatory diseases and their activity is mediated by flavonoids and lipophilic compounds.

Mexican oregano infusions have been traditionally used in Mexico for the treatment of inflammation-related diseases, such as respiratory and digestive disorders, headaches and rheumatism, among others. Nevertheless, there is limited information regarding the phenolic compounds, terpenes and composition as well as biological activity of Mexican oregano.To determine the phenolic and terpene composition and to evaluate the anti-inflammatory potential of three species of Mexican oregano (Lippia graveolens (LG), Lippia palmeri (LP) and Hedeoma patens (HP)) in order to provide a scientific basis for their use.We obtained methanol and chloroform extracts from dried oregano leaves of each species. We used LC-DAD-ESI-MS/MS and GC-MS to determine the phenolic and terpene profiles of the leaves, respectively. We evaluated anti-inflammatory potential by measuring the effect of Mexican oregano extracts on some pro-inflammatory mediators, such as nitric oxide (NO) and reactive oxygen species (ROS) using lipopolysaccharide(LPS)-stimulated RAW 264.7 macrophage cells and evaluating cyclooxygenase activity (COX-1, COX-2).Nine phenolic compounds (phenolic acids and flavonoids) and 22 terpenes (monoterpenes and sesquiterpenes) were detected in LG, LP and HP. We studied extracts from LG, LP and HP, and fractions from LG and LP in order to know their effect on some pro-inflammatory mediators. The phenolic and terpene extracts from LG, LP and HP exhibited significant inhibitory effect on ROS and NO production and mitochondrial activity in LPS-induced inflammation in RAW 264.7 macrophage cells. Nitric oxide production was also diminished by the terpene LG fraction LGF2 and the LP fractions LPF1, LPF2 and LPF3, confirming that both monoterpenes and sesquiterpenes are active compounds of oregano. Furthermore, the total extracts of LG, LP and HP exhibited non-selective inhibitions against the activity of the cyclooxygenases COX-1 and COX-2.Our results suggest that Lippia graveolens, Lippia palmeri and Hedeoma patens extracts have the potential to treat inflammatory diseases; their activity is mediated by polyphenols and terpenes. These findings support the claim for their traditional use in the treatment of inflammation-related diseases.