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Mycobacterium abscessus(MAB) is one of the rapidly growing, multidrug-resistant non-tuberculous mycobacteria (NTM) causing various diseases including pulmonary disorder. Although it has been known that type I interferons (IFNs) contribute to host defense against bacterial infections, the role of type I IFNs against MAB infection is still unclear. In the present study, we show that rIFN-β treatment reduced the intracellular growth of MAB in macrophages. Deficiency of IFN-α/β receptor (IFNAR) led to the reduction of nitric oxide (NO) production in MAB-infected macrophages. Consistently, rIFN-β treatment enhanced the expression of iNOS gene and protein, and NO production in response to MAB. We also found that NO is essential for the intracellular growth control of MAB within macrophages in an inhibitor assay using iNOS-deficient cells. In addition, pretreatment of rIFN-β before MAB infection in mice increased production of NO in the lungs at day 1 after infection and promoted the bacterial clearance at day 5. However, when alveolar macrophages were depleted by treatment of clodronate liposome, rIFN-β did not promote the bacterial clearance in the lungs. Moreover, we found that a cytosolic receptor nucleotide-binding oligomerization domain 2 (NOD2) is required for MAB-induced TANK binding kinase 1 (TBK1) phosphorylation and IFN-β gene expression in macrophages. Finally, increase in the bacterial loads caused by reduction of NO levels was reversed by rIFN-β treatment in the lungs of NOD2-deficient mice. Collectively, our findings suggest that type I IFNs act as an intermediator of NOD2-induced NO production in macrophages and thus contribute to host defense against MAB infection.

Sophora flavescens, also known as Kushen, has traditionally been used as a herbal medicine. In the present study we evaluated the ameliorative effects of kushenol C (KC) from S. flavescens against tBHP (tert-Butyl hydroperoxide)-induced oxidative stress in hepatocellular carcinoma (HEPG2) cells and acetaminophen (APAP)-induced hepatotoxicity in mice. KC pretreatment protected the HEPG2 cells against oxidative stress by reducing cell death, apoptosis and reactive oxygen species (ROS) generation. KC pretreatment also upregulated pro-caspase 3 and GSH (glutathione) as well as expression of 8-Oxoguanine DNA Glycosylase (OGG1) in the HEPG2 cells. The mechanism of action was partly related by KC’s activation of Akt (Protein kinase B (PKB)) and Nrf2 (Nuclear factor (erythroid-derived 2)-like 2) in the HepG2 cells. In in vivo investigations, coadministration of mice with KC and APAP significantly attenuated APAP-induced hepatotoxicity and liver damage, as the serum enzymatic activity of aspartate aminotransferase and alanine aminotransferase, as well as liver lipid peroxidation and cleaved caspase 3 expression, were reduced in APAP-treated mice. Coadministration with KC also up-regulated antioxidant enzyme expression and prevented the production of proinflammatory mediators in APAP-treated mice. Taken together, these results showed that KC treatment has potential as a therapeutic agent against liver injury through the suppression of oxidative stress.

Problem Chorioamnionitis is caused by a bacterial infection that ascends from the vagina and can cause adverse pregnancy outcomes (APOs). Fusobacterium nucleatum (F. nucleatum) is a periodontal pathogen associated with the occurrence of APOs. In this study, we evaluated whether receptor-interacting protein kinase 2 (Ripk2), an adaptor protein of the cytosolic receptors nucleotide-binding oligomerization domain (NOD)1 and NOD2, in macrophages and human decidual stromal cells (hDSCs) contributes to immune responses against F. nucleatum. Method of study Bone marrow-derived macrophages (BMDMs) isolated from wild-type (WT) and Ripk2-deficient mice and hDSCs were cultured with F. nucleatum (MOI 1, 10, 100). BMDMs and hDSCs were assessed using enzyme-linked immunosorbent assay, Western blot analysis, real-time PCR, and nitrite assay. Results Fusobacterium nucleatum-induced production of IL-6, but not of TNF-α and IL-10, was lower in Ripk2-deficient BMDMs than in WT cells. Western blotting revealed a decrease in F. nucleatum-induced p65 phosphorylation in Ripk2-deficient macrophages, whereas mitogen-activated protein kinases activation was comparable between WT and Ripk2-deficient cells. The production of nitric oxide (NO) in response to F. nucleatum and the gene and protein expression of inducible NO synthase was impaired in Ripk2-deficient BMDMs. In hDSCs, F. nucleatum upregulated the gene and protein expression of NOD1, NOD2, and Ripk2 in a time-dependent manner. F. nucleatum also increased the production of IL-6, CXCL8, and CCL2, whereas this production was decreased by the Ripk2 inhibitors SB203580 and PP2. Conclusions In conclusion, Ripk2 signaling appears to contribute to the F. nucleatum-induced immune response and can be a preventive and therapeutic target against APOs.

A novel compound 1 and nine known compounds (2–10) were isolated by open column chromatography analysis of the root bark of Ulmus davidiana. Pure compounds (1–10) were tested in vitro to determine the inhibitory activity of the catalytic reaction of soluble epoxide hydrolase (sEH). Compounds 1, 2, 4, 6–8, and 10 had IC50 values ranging from 11.4 ± 2.3 to 36.9 ± 2.6 μM. We used molecular docking to simulate inhibitor binding of each compound and estimated the binding pose of the catalytic site of sEH. From this analysis, the compound 2 was revealed to be a potential inhibitor of sEH in vitro and in silico. Additionally, molecular dynamics (MD) study was performed to find detailed interaction signals of inhibitor 2 with enzyme. Finally, compound 2 is promising candidates for the development of a new sEH inhibitor from natural plants.

Four Korea-native wild vegetables belonging to the Allium genus (Allium tuberosum Rottl., Allium senescens L., Allium thunbergii G. Don., and Allium sacculiferum Maxim.) were investigated for their antioxidant and anti-obesity potential, in order to promote the utilization of wild edible plants for the development of potential new crops. In this study, we compared the antioxidant activity of the plant extracts by ABTS+ and DPPH scavenging method. We then compared the anti-adipogenic effects of the extracts in adipocytes. We quantified the phenolics in the extracts using high-performance liquid chromatography (HPLC) to identify the bioactive compounds. A. tuberosum and A. sacculiferum had a higher total phenolic content (19.0 ± 0.3 and 19.6 ± 0.7 mg GA/g extract, respectively) than A. senescens and A. thunbergii (16.0 ± 0.2 and 14.5 ± 0.7 mg GA/g extract). Caffeic acid, an antioxidant and anti-adipogenic compound, was detected in A. tuberosum and A. sacculiferum, but not in A. senescens and A. thunbergii. A. tuberosum, and A. sacculiferum showed superior antioxidative and anti-adipogenic activities compared to A. senescens and A. thunbergii. Therefore, A. tuberosum and A. sacculiferum, both of which contain antioxidant and anti-adipogenic phenolics, such as caffeic acid, present a therapeutic potential against obesity. Various phenolic compounds, which have been found to differ among Allium spp., may partially account for differences among the Allium spp. with respect to their antioxidant and anti-obesity effects.

Purpose: To introduce a case of corneal endothelial toxicity caused by Asclepias curassavica (Milkweed) in Korea. Observations: A 37-year-old Asian man presented with decreased vision and redness in the right eye, which developed after contact with Asclepias curassavica. At presentation, best-corrected visual acuity (BCVA) was 20/60 in the right eye. Slit lamp examination demonstrated severe corneal stromal edema with Descemet's folds and conjunctival hyperemia. We prescribed topical prednisolone acetate 1% eye drops (8 times a day), cyclosporine 0.1% (once a day) and oral prednisolone (30 mg a day for 3 days). One day later, the BCVA improved to20/40 and marked improvement in corneal edema was observed. At 5 days, BCVA was 20/22 and anterior segment examination showed minimal corneal edema with resolution of Descemet's folds. At 2 weeks, BCVA was 20/20 in the right eye and corneal edema completely resolved. Conlcusions and importance: This case suggests that high index of suspicion for toxicity from Asclepias species is necessary when encountered with patients who present with corneal edema after exposure to these plants. Aggressive anti-inflammatory treatment might be helpful for early recovery, at least for young patients. Keywords: Asclepias, Corneal edema, Corneal endothelial dysfunction, Inflammation, Millkweed

Background: The root of Cynanchum auriculatum Royle ex Wight has been traditionally used as a folk medicine in the eastern Asia. Objectives: In this study, we explored the immune-enhancing effect of the high-molecular weight fraction of C. auriculatum (CAHF) root extracts in RAW 264.7 macrophages and cyclophosphamide (CYP)-induced immune-suppressed mice. Materials and Methods: To obtain the CAHF, ultrafiltration was performed using 30-kDa molecular weight cutoff polyethersulfone membranes in a cross-flow microfiltration system. In vitro and in vivo experiments were performed to evaluate the immunostimulatory effects of CAHF. Results: CAHF increased the production of nitric oxide and immunostimulatory cytokines (interleukin-6 and tumor necrosis factor-α) in recombinant interferon-γ primed macrophages. In addition, the phagocytic activity of macrophage cells was enhanced by CAHF. Furthermore, CAHF treatment markedly restored the decreased body weight, immune organs (spleen and thymus) weight, natural killer cell activity, and T- and B-lymphocyte proliferation in CYP-induced immune-suppressed mice. Conclusion: These results suggest that CAHF is an effective immunostimulatory agent.

Background : The aim of the present study was to identify an effective physicochemical control method to reduce Fusarium species infestation in adlay (Coix lacryma-jobi L.) before and after harvesting. Methods and Results : We observed that prochloraz emusifiable concentrate and hexaconazol prochloraz emusifiable concentrate strongly inhibited the mycelial growth of 10 Fusarium species. Strong growth inhibitions and cell lysis were observed following treatment with 4% NaOCl solution. The total number of fungi detected were lower follwing treatment with thiophanatemethyl triflumizole wettable powder (), hexaconazol prochloraz emulsifiable concentrate (), carboxin thiram dustable powder () and prochloraz emulsifiable concentrate () than in the non-treated control (). The reduction of Fusarium fungi varies with the concentration and soaking time of NaOCl solution. Fungal detection was not observed after soaking in NaOCl solution for 24 h and harmful effects were not observed for plant growth by NaOCl after soacking for 6 - 12 h. Conclusion : Soaking seed for 6 - 12 h in 4% NaOCl could be an effective method of disinfectant treatment for the control of Fusarium fungi in adlay seeds.

*Department of Herbal Crop Research, NIHHS, RDA, Eumsung 369-873, Korea.**Department of Plant Resources, College of Industrial Science, Kongju National University, Yesan 340-702, Korea.ABSTRACT : This study was conducted to obtain the basic data related to seed characteristics and germination conditionsof the Coriandrum sativum L. at different storage condition and temperature. The shape of fruit was oval with light browncolor. Fruit was mericarp, biloculate, with one ovule in each locus. The length and width of seed were 1.37±0.067 ㎜ and0.52 ±0.039 ㎜, respectively. Weight of 1,000 seeds was 6.55±0.15 g. Seeds which were stored at room (15℃) and cold tem-perature (4℃) in vinyl container showed the highest germination rate (93.3%) under the room temperature germinationcondition. Percentage of germination in cold and freeze temperature was 20% and 0%, respectively. Germination rate ofroom storage seeds with paper container increased to about 91.3%, however, decreased in cold (4℃) and freeze storage (−20℃)with paper container (76.7% and 78.0%, respectively). Germination rate of seed in vinyl stock container was better thanthat of paper stock container. Germination rate of seeds stored at room temperature for 4 years (2010 - 2013) ranged from80.0% to 91.3%. Therefore, coriander seeds are expected to be stored at room temperature for increasing the germinationrate and keeping a long term.Key Words : Coriandrum sativum L., Germination Temperature, Seed Characteristics, Storage Condition

The objective of this study was to investigate the immune-enhancing activity of a high molecular weight fraction (HMF) of Cynanchum wilfordii in RAW 264.7 macrophages and the cyclophosphamide (CYC)-induced mouse model of immunosuppression. To identify the bioactive substances of HMF, a crude polysaccharide (HMFO) was obtained and treated with sodium periodate (an oxidation agent) or digested with protease. In macrophages, HMF treatment enhanced the production of nitric oxide (NO) and cytokines (tumor necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), and interleukin 1 beta (IL-1 beta)), as well as phagocytic ability. In CYC-immunosuppressed mice, HMF improved relative spleen and thymus weights, natural killer (NK) cell activity, and splenic lymphocyte proliferation. These increases in NO and cytokines were mediated by up-regulation of nuclear factor kappa B (NF-kappa B) and mitogen-activated protein kinase (MAPK) signaling pathways. Periodate treatment, but not protease treatment, decreased the immune-enhancing activity of HMFO, suggesting that polysaccharides are the active ingredients in C. wilfordii extract.

Oligonucleotides that carry a detectable label can be used to probe for mRNA targets in in situ hybridization experiments. Oligonucleotide probes (OPs) have several advantages over cDNA probes and riboprobes. These include the easy synthesis of large quantities of probe, superior penetration of probe into cells and tissues, and the ability to design gene- or allele-specific probes. One significant disadvantage of OPs is poor sensitivity, in part due to the constraints of adding and subsequently detecting multiple labels per oligonucleotide. In this study, we compared OPs labeled with multiple detectable haptens (such as biotin, digoxigenin, or fluorescein) to those directly conjugated with horseradish peroxidase (HRP). We used branching phosphoramidites to add from two to 64 haptens per OP and show that in cells, 16-32 haptens per OP give the best detection sensitivity for mRNA targets. OPs were also made by directly conjugating the same oligonucleotide sequences to HRP. In general, the HRP-conjugated OPs were more sensitive than the multihapten versions of the same sequence. Both probe designs work well both on cells and on formaldehyde-fixed, paraffin-embedded tissues. We also show that a cocktail of OPs further increases sensitivity and that OPs can be designed to detect specific members of a gene family. This work demonstrates that multihapten-labeled and HRP-conjugated OPs are sensitive and specific and can make superior in situ hybridization probes for both research and diagnostic applications.

The plasma membrane-localized BRASSINOSTEROID-INSENSITIVE1 (BRI1) and BRI1-ASSOCIATED KINASE1 (BAK1) are a well-known receptor pair involved in brassinosteroids (BR) signaling in Arabidposis. The formation of a receptor complex in response to BRs and the subsequent activation of cytoplasmic domain kinase activity share mechanistic characteristics with animal receptor kinases. Here, we demonstrate that BRI1 and BAK1 are BR-dependently phosphorylated, and that phosphorylated forms of the two proteins persist for different lengths of time. Mutations of either protein abolished phosphorylation of the counterpart protein, implying transphosphorylation of the receptor kinases. To investigate the specific amino acids critical for formation of the receptor complex and activation of BAK1 kinase activity, we expressed several versions of BAK1 in yeast and plants. L32E and L46E substitutions resulted in a loss of binding of BAK1 to BRI1, and threonine T455 was essential for the kinase activity of BAK1 in yeast. Transgenic bri1 mutant plants overexpressing BAK1(L46E) displayed reduced apical dominance and seed development. In addition, transgenic wild type plants overexpressing BAK1(T455A) lost the phosphorylation activity normally exhibited in response to BL, leading to semi-dwarfism. These results suggest that BAK1 is a critical component regulating the duration of BR efficacy, even though it cannot directly bind BRs in plants.