Search

Optimal transport conditions for the live black rockfish and changes in their characteristics during long-distance transport were investigated. Oxygen consumption by black rockfish was lowe...

Context:Allium senescens Linn. (Liliaceae) (ASL) has been traditionally used in Korea and other Asian countries for improving digestive and liver functions. Objective: The anti-hepatofibrosis effect of ASL ethanol extract in cellular and experimental fibrosis rat model was investigated. Materials and methods:In vitro cell viability, cell cycle and apoptosis in hepatic stellate cells (HSCs) were studied using MTT assay, flow cytometry and Annexin V-FITC/PI staining. Thioacetamide (TAA; 200 mg/kg, i.p.)-induced liver fibrosis model using Sprague Dawley rats (n = 10) was developed in vivo by injecting TAA twice per week for 13 weeks. ASL (25 and 100 mg/kg) and silymarin (50 mg/kg) were administered through oral gavage 2 times per week from 7th to 13th week. Specific fibrotic-related biomarkers such as aspartate transaminase (AST), alanine transaminase (ALT), glutathione and hydroxyproline levels in serum were analyzed by spectrophotometer using commercial kits. Morphological, histopathological and fibrotic-related gene expression such as TGF-β, Col1α1 and α-SMA in liver tissues was estimated by hematoxylin and eosin staining, Picrosirius red stain and quantitative real-time polymerase chain reaction, respectively. Results: ASL (0.1 mg/mL) and silymarin (0.05 mg/mL) treatment induced apoptosis (4.06% and 8.67%) in activated HSC-T6 cells, compared with control group (3.7%). The altered morphology in activated primary HSCs was also restored by ASL (0.1 mg/mL) treatment. Further, ASL (100 and 25 mg/kg) ameliorated the TAA-induced altered fibrotic-related biomarkers, histopathological changes and fibrotic-related gene expression significantly (p

There are many types of scars. They vary according to the shape of the scar surface, shape of the scar itself, color, and chronology. Scars may be classified by scar surface (flat, elevated, depressed, or irregular) by morphology (atrophic, normorophic, hypertrophic, or keloid) by color (depigmented, hypopigmented, hyperpigmented, or discolored) or by chronology (acute, chronic, immature, or mature) [1].

Context: Ampelopsis brevipedunculata Maxim (Vitaceae) is a traditional medicinal herb used for treating liver disorders. Objective: The hepatoprotective effects of A. brevipedunculata ethanol extract (ABE) was investigated in experimental models of fibrosis. Materials and methods: Hepatic stellate cells (HSCs) system in vitro and thioacetamide (TAA)-induced liver fibrosis rat model in vivo were used. Sprague–Dawley rats were divided into five groups of eight each (control, TAA, TAA with ABE 10 mg/kg, ABE 100 mg/kg and silymarin 50 mg/kg groups, respectively). Fibrosis was induced except to the control group by TAA (200 mg/kg, i.p.) twice per week for 13 weeks. ABE and silymarin was administered orally six times per week from the 7th week to the 13th week. Results: In HSC-T6 cells, ABE (0.1 mg/mL) and silymarin (0.05 mg/mL) significantly (p

This study was performed to obtain basic data regarding the development of fish steak products using olive flounder and pork leg. Olive flounder and pork leg were ground separately in a chopper. The methods used for processing were as follows. Chopped olive flounder (100 g) and other ingredients (bread crumbs, 13 g; onion, 12 g; garlic, 4 g; egg wash, 18 g; salt, 0.05 g; pepper, 0.05 g) were mixed in a chopper. The mixture was molded into a steak shape () and roasted in an oven at for 12 min (OF). FP consisting of a mixture of olive flounder (70 g) and pork leg meat (30 g) and OP consisting of pork leg meat alone (100 g) were processed according to the same procedure as described for OF. Various factors (viable bacterial count, chemical composition, pH, salinity, hardness value, color value, total amino acid content, free amino acid content, fatty acid composition, mineral content) were measured, and sensory evaluation was conducted. Based on the results of the sensory evaluation and hardness value, OP was deemed to be the most desirable, followed in order by FP and OF. There was a slight but significant difference between OP and FP.

Rhus javanica Linn, a traditional medicinal herb from the family Anacardiaceae, has been used in the treatment of liver diseases, cancer, parasitic infections, malaria and respiratory diseases in China, Korea and other Asian countries for centuries. In the present study, the protective effects of R. javanica ethanolic extract (RJE) on hydrogen peroxide (H2O2)-induced oxidative stress in human Chang liver cells was investigated. The cell cytotoxicity and viability were assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The activities of superoxide dismutase (SOD) and catalase (CAT) were measured using respective enzymatic kits. Cell cycle analysis was performed using flow cytometric analysis. The protein expression levels of p53, B-cell lymphoma (Bcl)-2, Bcl-2-associated X protein (Bax) and caspase-3 were assessed by western blotting. Human Chang liver cells were treated with different concentrations (0.1, 0.3 or 0.5 mg/ml) of RJE, and were subsequently exposed to H2O2 (30 µM). Treatment with H2O2 (30 µM) significantly induced cytotoxicity (P

This study was undertaken to investigate the quality changes of salted-fermented anchovy meat made by varying the amount of salt during fermentation at 15℃. Anchovy (11.0-14.0 cm of length, 10.7-17.5 g of weight) added with 15-25% of salt was filled in a round form plastic container (i.d. 10.5x11 cm), and then fermented at 15℃ for 110 days. The factors such as proximate composition, pH, color value (L, a, b), TBA value, amino-N content, salinity, hardness value, free amino acid content and sensory evaluation of salted-fermented anchovy meat were measured. Ash content, color value (redness), TBA value, amino-N content. salinity and hardness value of salted-fermented anchovy meat were increased, but color value (lightness), and moisture content were decreased during fermentation at 15℃. A salted-fermented anchovy meat added with 15% of salt was shown higher content of moisture, amino-N content and free amino acid, TBA value than those of 20 or 25% of salt. Ash content, salinity and hardness value were highest in a product added with 25% of salt. From the result of sensory evaluation, Addition amount of 15% salt and fermentation periods of 110 days were determined to be the most desirable palatability of salted-fermented anchovy meat. Key words : Anchovy, Salt concentration, Fermenting temperature, Salted-fermented anchovy meatCorresponding author : 055-772-9141, kimjg@gnu.ac.kr

Background: This study aims to evaluate the efficacy, safety, and appropriate dose of Hanslim, a Korean traditional herbal medicine, for obese patients, when compared to a placebo. Methods/design: This study is a randomized, double-blinded, multicenter, multidose, placebo-controlled, phase IIb clinical trial. A total of 165 obese patients with a body mass index (BMI) of more than 30 kg/m2 or obese patients with a BMI of 27 to 29.9 kg/m2 and one or more risk factors such as hypertension, diabetes, or hyperlipidemia will be enrolled. Participants will be randomly assigned to 1 of 3 groups (high-dose, low-dose, or placebo) with a 1:1:1 allocation ratio and will have 4 scheduled visits during the 12-week treatment period. The participants will be administered 2 tablets of Hanslim or placebo, 2 times per day. The difference in the proportion of participants who lost weight by more than 5% from their baseline at 12 weeks compared to the placebo group will be examined as the primary efficacy outcome. Secondary efficacy outcomes include differences in body weight, BMI, body-fat percentage, fat mass, skeletal-muscle mass, edema index, waist circumference, hip circumference, waist-hip ratio, serum lipid, blood glucose, C-reactive protein, and total score of Korean version of obesity-related quality of life after 12 weeks of treatment. Adverse events, laboratory test results, vital sings, and electrocardiography will be recorded to evaluate safety. Discussion: This is the first prospective clinical trial to explore the efficacy and safety of Hanslim for obese patients. If the results provide the appropriate dosage of Hanslim, this study would contribute to the confirmatory evidence for the use of Hanslim as a treatment for obesity needed to conduct a large-scale, phase III clinical trial. The results will be published in a peer-reviewed journal. Trial registration: Clinical Research Information Service, ID: KCT0002193. Registered on January 6, 2017. https://cris.nih.go.kr/cris/search/search_result_st01_en.jsp?seq=7468

Aquaculture of olive flounder started in the middle of 1980's and now farming has been taken place in many places along the coastal line in Korea. The taste of olive flounder has a good chewy texture because of high collagen content, low fat content, so it is popular for sliced raw fish. Olive flounder is popular among Koreans but the consumption pattern is uniformly so as to be used as sliced raw fish but not other ways. So, now there needs to develop high valued-processed food using olive flounder.This study was set to investigate the processing of terrine by using olive flounder, in which terrine is French style meat loaf that is well favored around the world. In this study, terrine was prepared by chopping olive flounder meat with 39 g egg white and 10 mL fresh cream (per 50 g fillet) and then seasoned with 5 mL lemon juice, 5 mL brandy, 0.05 g salt and 0.05 g pepper. The 25 g of dough was placed on a vinyl wrap, put with 2 g cheese, and layered an another 25 g dough, and then rolled up and wrapped by aluminium foil. Two different cooking methods were used for terrine processing in this study. Terrine-1 was cooked by vacuum sealed in polyethylene film (20×30×0.05 mm) after boiling for 5 min and stored at -20℃ for 7 days. Terrine-2 was prepared by vacuum sealed in polyethylene film (20×30×0.05 mm) and stored at -20℃ for 7 days. After 7 days, Terrine-1 was thawed and then heated up in microwave for 2 min (Sample-1), while Terrine-2 was thawed and then boiled in water for 5 min (Sample-2). Viable bacterial count, chemical composition, pH, salinity, hardness, TBA, free amino acid content, and sensory evaluation were measured for both Sample-1 and Sample-2. Especially, the scores of sensory evaluation of Sample-2 is slightly higher than that of Sample-1. On the other hand, there were no significant differences on color, odor, taste, texture, and overall acceptance between Sample-1 and Sample-2.Key words : Terrine, Olive flounder, Sensory evaluation, BoilingCorresponding author : 055-772-9141, kimjeonggyun@nate.com

Pachymic acid (PA) is a lanostane-type triterpenoid derived from Poria cocos mushroom that possess various biological effects such as anti-cancer, antiinflammatory and anti-metastasis effects. In this study, we investigated the anti-cancer effects of PA in EJ bladder cancer cells. The results showed that PA significantly inhibited proliferation of EJ cells in a dose-dependent manner. PA induced accumulation of sub-G1 DNA content (apoptotic cell population), apoptotic bodies and chromatin condensation and DNA fragmentation in EJ cells in a dose-dependent manner. PA also induces activation of caspase-3, -8 and -9, and subsequent cleavage of poly (ADP-ribose) polymerase, and significantly suppressed the inhibitor of apoptosis protein family proteins in a dose-dependent manner. Furthermore, PA activates Bid and induced the loss of mitochondrial membrane potential (ΔΨm ) with up-regulated pro-apoptotic proteins (Bax and Bad), down-regulated anti-apoptotic proteins (Bcl-2 and Bcl-xL) and cytochrome c release. In turn, PA increased the generation of reactive oxygen species (ROS); also, the ROS production was blocked by N-acetyl-L-cysteine. The expressions of TNF-related apoptosis inducing ligand and death receptor 5 were up-regulated by PA in a dose-dependent manner, suggesting extrinsic pathway also involved in PA-induced apoptosis. This study provides evidence that PA might be useful in the treatment of human bladder cancer.

Pachymic acid (PA) is a lanostane-type triterpenoid derived from the Poria cocos mushroom. Several beneficial biological features of PA provide medicine with a wide variety of valuable effects, such as anticancer and anti-inflammatory activity; it also has antioxidant effects against oxidative stress. Nonetheless, the biological properties and mechanisms that produce this anti-cancer action of PA remain largely undetermined. In this study, we investigated the pro-apoptotic effects of PA in T24 human bladder cancer cells. It was found that PA could inhibit the cell growth of T24 cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies and chromatin condensation and accumulation of cells in the sub-G1 phase. The induction of apoptotic cell death by PA was connected with an up-regulation of pro-apoptotic Bax and Bad protein expression and down-regulation of anti-apoptotic Bcl-2 and Bcl-xL proteins, and inhibition of apoptosis family proteins. In addition, apoptosis-inducing concentrations of PA induced the activation of caspase-9, an initiator caspase of the mitochondrial-mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose)-polymerase. PA also induced apoptosis via a death receptor-mediated extrinsic pathway by caspase-8 activation, resulting in the truncation of Bid and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. Taken together, the present results suggest that PA may be a potential chemotherapeutic agent for the control of human bladder cancer cells.

Oysters, especially are excellent source of several mineral including iron, zinc and selenium, which are often low in the mordern diet. They are also an excellent source of glycogen, vitamin B12 and considered the healthiest when eaten raw on the half shell in good tasted season from November to March.This study was investigated for the purpose of obtaining basic data which can be applied to processing of two kinds of canned boiled oyster (canned boiled oyster, canned boiled oyster in bamboo salt). Shucked oyster meat was cooked in steam (15min) after washing with water, filled 90g into can (301-3), added with salt solution and then precooked for 10 min. at 100℃. Canned boiled oyster was added 1.5% salt solution 60mL. Canned boiled oyster in bamboo salt was added 0.5% salt solution 30mL and 0.7% bamboo salt solution 30mL. The cans were seamed using a vacuum seamer, and then sterilized for various Fo values (Fo 8~12 min.) in a steam system retort at 116℃, 118℃. Viable bacterial count, proximate composition, pH, salinity, yield, VBN, amino-N, TBA, mineral, color value, free amino acid, hardness and sensory evaluation of two kinds of canned boiled oyster produced at various sterilization condition (Fo 8~12 min.) were measured after divide to meat and juice. The results showed that canned boiled oyster in bamboo salt sterilized at Fo 8 min. was the most desirable because this condition is the most economical and tasty.Key words : Oyster, Canned boiled oyster, Canned boiled oyster in bamboo salt, Sterilization, Fo value Corresponding author : 055-772-9141, kimjeonggyun@nate.com

To develop an effective use for poor-quality individually quick-frozen (IQF) oysters Crassostrea gigas stored for a long period, the extract conditions, quality characteristics, and optimum reaction flavoring (RF) conditions of a complex extract from these IQF oysters were investigated. The moisture, pH, and volatile basic nitrogen contents of IQF oysters stored for 18 months (18M-IQFO) were 77.9%, 6.32, and 17.9 mg/100 g, respectively. Three different kinds of extract were prepared from 18M-IQFO: a hot-water extract (HE), scrap enzymatic hydrolysate (EH), and complex extract (CE). The respective extracts contained 5.5, 8.6, and 6.6% crude protein and 281.7, 366.0, and 343.0 mg/100 g amino nitrogen, and had 811, 359, and 1,170 mL/kg extraction yields. The CE was superior to the traditional HE in terms of the extraction yield, amino-nitrogen content, and organoleptic qualities, except for the odor. To improve flavor via the Maillard reaction, the reaction system used to produce a desirable flavor comprised CE (Brix 30°), 0.4 M glucose, 0.4 M glycine, and 0.4 M cysteine solution (4:2:1:1, v/v). The reaction time and pH were the independent variables, and the sensory scores for baked potato odor, masking shellfish odor, and boiled meat odor were the dependent variables. The surface response methodology (RSM) analysis of the multiple responses optimization gave a reaction time of 120.6 minutes and pH 7.33 at 120°C. The reaction improved the flavor of CE considerably, as compared to that of the unreacted extract.

과메기는 조선시대 부터 생선의 보존성을 향상시키기 위해 경 북 동해안 일대에서 건조시켜서 만든 대표적인 전통·향토식품 으로 알려져 있다(Oh and Kim, 1995). 과메기는 1960년 이전 에는 주로 청어를 동절기에 자연건조 하여 만들었으나 어획량 의 감소와 기온의 상승으로 최근에는 청어보다 건조가 용이한 꽁치를 이용하여 제조하고 있다(Oh and Kim, 1998; Oh et al., 1998a). 꽁치나 청어는 고등어, 방어, 정어리, 전갱이, 다랑어 등 과 함께 등푸른 생선으로 불리며, 고도불포화지방산인 eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) 함량이 높아 심근경색, 뇌경색, 고혈압 등 주요 성인병 예방의 생리적 기능을 가지고 있는 것으로 알려져 있다(Oh et al., 1998a). 또 한 어린이 성장촉진 및 두뇌발달에 효과적인 성분과 여성피부 에 효과가 있는 성분이 다량 들어 있다는 연구가 발표되어 있다 (Uhei et al., 1990). 과메기에 관하여 1995년 이후부터 현재까지 많은 발표 논문 이 발표되어 있다. 즉, 과메기의 제조조건에 관한 연구로는 Oh and Kim (1995)이 꽁치 자연동결건조(과메기) 중 지방함량과 지방산조성의 변화, Oh et al. (1998a)이 과메기 제조 시 건조 조건에 따른 꽁치 근육의 성분변화, Oh et al. (1998b)이 꽁치 과메기의 건조조건에 따른 amine의 변화, Oh and Kim (1998) 이 핵산류 및 유리아미노산의 변화, Oh et al. (1996)이 콜레스 테롤 함량 변화에 대하여 보고하였으며, 과메기의 저장조건에 초고추장첨가 과메기통조림의 제조 및 특성

Context: Cuscuta chinensis Lam. (Convolvulaceae) has been used as a traditional herbal remedy for treating liver and kidney disorders. Objective: Anti-fibrotic effects of C. chinensis extract (CCE) in cellular and experimental animal models were investigated. Materials and methods: HSC-T6 cell viability, cell cycle and apoptosis were analysed using MTT assay, flow cytometry and Annexin V-FITC/PI staining techniques. Thioacetamide (TAA)-induced fibrosis model was established using Sprague Dawley rats (n = 10). Control, TAA, CCE 10 (TAA with CCE 10 mg/kg), CCE 100 (TAA with CCE 100 mg/kg) and silymarin (TAA with silymarin 50 mg/kg). Fibrosis was induced by TAA (200 mg/kg, i.p.) twice per week for 13 weeks. CCE and silymarin were administered orally two times per week from the 7th to 13th week. Fibrotic related gene expression (α-SMA, Col1α1 and TGF-β1) was measured by RT-PCR. Serum biomarkers, glutathione (GSH) and hydroxyproline were estimated by spectrophotometer using commercial kits. Results: CCE (0.05 and 0.1 mg/mL) and silymarin (0.05 mg/mL) treatment significantly (p

Jasin-hwan-gagambang (BHH10), a modified prescription of Jasin-hwan, contains Astragalus membranaceus, Cinnamomum cassia, and Phellodendron amurense, and it has been traditionally used to treat osteoporosis and other inflammatory diseases. In this study, we systematically investigated the protective effects of BHH10 in ovariectomy (OVX)-induced rats. Sprague–Dawley rats were randomly divided into sham and OVX subgroups. The rats in the OVX group were treated with vehicle, BHH10, alendronate (ALN), and 17β-estradiol (E2). BHH10 treatment significantly inhibited OVX-induced increases in body weight and uterus atrophy. In addition, it significantly increased the bone mineral density (BMD) and prevented a decrease in trabecular bone volume, connectivity density, trabecular number, thickness, and separation at the total femur and femur neck. The OVX rats showed significant decreases in the serum levels of calcium and phosphorous and significant increases in the serum levels of cholesterol, low-density lipoprotein cholesterol, alkaline phosphatase, osteocalcin, C-telopeptide type 1 collagen, and bone morphogenetic protein-2. These changes were significantly reduced to near sham levels by administration of BHH10 to OVX rats. BHH10-treated rats had a greater bone mass, a better structural architecture of the bone, and higher levels of biochemical markers of the bone than did the ALN-treated or E2-treated rats. These results suggest that BHH10 reverses osteoporosis in OVX rats by stimulating bone formation or regulating bone resorption and is not associated with toxicity. © 2014 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.

This study was conducted to explore the antiadipogenic effect and possible mechanism of Gambisan on 3T3-L1 cells. For quality control, Gambisan was standardized by HPLC and the standard compounds ephedrine, epigallocatechin-3-gallate, and caffeine were screened. Cultured 3T3-L1 cells that had been induced to differentiate were treated with various concentrations of Gambisan or its major component extracts (Ephedra intermediaSchrenk,Atractylodes lanceaDC., andThea sinensisL.) for 72 hours for MTT assay to determine cell viability or 10 days for LDH assay, triglyceride assay, DNA content measurement, Oil red O staining, RT-PCR, and western blot. Gambisan significantly inhibited adipogenesis in 3T3-L1 cells by reducing triglyceride contents and lipid accumulation in a dose-dependent manner without obvious cytotoxicity. Viability and DNA content in 3T3-L1 cells treated with Gambisan were significantly higher than cells treated with the major component extracts at every concentration. The anti-adipogenic effects of Gambisan appeared to be mediated by a significant downregulation of the expression of lipoprotein lipase mRNA and PPARγ, C/EBPα, and SREBP-1 protein apart from the expression of hormone-sensitive lipase. Gambisan could act as a possible therapeutic agent for obesity. However, further studies includingin vivoassays and clinical trials are needed to confirm the efficacy, safety and mechanisms of the antiobesity effects of Gambisan.

We investigated the effect of galangin, a natural flavonoid, on osteoclastic bone destruction in collagen-induced arthritis and examined the molecular mechanisms by which galangin affects osteoclastogenesis in bone marrow derived macrophages. In mice with collagen-induced arthritis, administration of galangin significantly reduced the arthritis clinical score, edema and severity of disease without toxicity. Interestingly, galangin treatment during a later stage of collagen-induced arthritis, using mice with a higher clinical arthritis score, still significantly slowed the progression of the disease. Extensive cartilage and bone erosive changes as well as synovial inflammation, synovial hyperplasia and pannus formation were dramatically inhibited in arthritic mice treated with galangin. Furthermore, galangin-treated arthritic mice showed a significant reduction in the concentrations of IL-1β, TNF-α and IL-17. We found that galangin inhibited osteoclastogenic factors and osteoclast formation in bone marrow-derived macrophages and osteoblast co-cultured cells, and increased osteoprotegerin (OPG) levels in osteoblasts. Galangin and NF-κB siRNA suppressed RANKL-induced phosphorylation of the c-jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK), but not AKT and extracellular signal-regulated kinase 1/2 (ERK1/2). Also, the JNK inhibitor SP600125 and p38 inhibitor SB203580 reduced RANKL-induced expressions of phospho-c-Jun, c-fos and NFATc1 genes during osteoclast development. In addition, galangin suppressed RANKL-induced phosphorylation of NF-κB, phospho-IκBα, inflammatory cytokines and osteoclast formation in bone marrow-derived macrophages. Our data suggest that galangin prevented osteoclastic bone destruction and osteoclastogenesis in osteoclast precursors as well as in collagen-induced arthritis mice without toxicity via attenuation of RANKL-induced activation of JNK, p38 and NF-κB pathways.

Melittin (1) is a major polypeptide in honey bee venom that has been used traditionally against chronic inflammation and cancer. However, its molecular mechanism has not been determined. In this study, the antitumor effect of 1 was compared with that of NS398, a cyclooxygenase-2 (COX-2) inhibitor, in vivo and in vitro. Subcutaneous injection of 1 at 0.5 and 5 mg/kg suppressed significantly vascular endothelial growth factor (VEGF)-A-transfected highly metastatic Lewis lung cancer (VEGF-A-hm LLC) tumor growth by 25% and 57%, respectively. Also, 1 inhibited significantly the number of vessels around VEGF-A-hm LLC cells. The results were superior to those obtained in the mice treated with NS398. Compound 1 dose-dependently inhibited proliferation and tube formation in human umbilical vein endothelial cells (VEGF-A-HUVECs), without affecting cell viability in native HUVECs. In addition, 1 decreased the expression of VEGF receptor-2 (VEGFR-2), COX-2, and prostaglandin E2 (PGE2) in VEGF-A-transfected HUVECs. These effects were accompanied by a reduction of the phosphorylation of extracellular signal-regulated kinase 1/2 and c-jun N-terminal kinase, whereas it increased the phosphorylation of p38 mitogen-activated protein kinase (MAPK). SB203580 abolished the downregulation of COX-2 and VEGFR-2 and the inhibition of cell proliferation by 1. The antitumor activity of 1 may be associated with antiangiogenic actions via inhibiting VEGFR-2 and inflammatory mediators involved in the MAPK signaling pathway.

Ethnopharmacological relevance The dried flowers of Lonicera japonica Thunb and dried roots of Anemarrhena asphodeloides BUNGE have been used for the treatment of a variety of inflammatory diseases in traditional Korean medicine. Objective The aim of the study is to evaluate the anti-inflammatory effects of WIN-34B, a new herbal medicine, in fibroblast-like synoviocytes (FLS) obtained from patients with osteoarthritis (OA). Materials and methods WIN-34B is isolated from the n-butanol fraction of dried flowers of L. japonica and dried roots of A. asphodeloides. The anti-inflammatory effects of WIN-34B on cell viability, the production and release of inflammatory mediators, matrix metalloproteinases (MMPs), aggrecanases, tissue inhibitor of matrix proteinases (TIMP) is compared with celecoxib in IL-1β-stimulated human OA FLS. Furthermore, the effect of WIN-34B on inhibitory kappa B-α (IκB-α) phosphorylation and mitogen-activated protein kinases (MAPK) in the IL-1β-stimulated OA FLS was also evaluated. Results WIN-34B significantly inhibited the IL-1β-induced cell viability in human OA FLS without cytotoxicity. Compared to celecoxib, WIN-34B exhibited similar or better anti-inflammatory effects through significant suppression of inflammatory mediators (IL-1β, TNF-α, PGE2 and NO), MMPs (MMP-1, MMP-3 and MMP-13) and aggrecanases (ADAMTS-4 and ADAMTS-5), and enhancement of TIMPs (TIMP-1 and TIMP-3). Moreover, WIN-34B reduced the phosphorylation of IκB-α, ERK1/2, p38 and JNK1/2 in IL-1β-stimulated OA FLS. Conclusions WIN-34B exhibited similar or better anti-inflammatory properties in IL-1β-stimulated OA FLS compared to celecoxib. The anti-inflammatory effects of WIN-34B are due to inhibition of inflammatory mediators (IL-1β, TNF-α, PGE2 and NO) and regulation of MMPs, ADAMTSs and TIMPs via the inhibition of IκB-α and MAPK phosphorylation in IL-1β-stimulated OA FLS.

Cystic fibrosis transmembrane conductance regulator (CFTR) gene and sodium-independent Cl?/HCO₃? anion exchanger (AE) genes are expressed in a wide variety of mammalian tissues including cholangiocytes. They play an important role in the regulation of intracellular pH (pHi) as well as in transepithelial acid/base transport necessary for biliary bicarbonate secretion. The aim of this study was to examine the expression level of CFTR gene and AE genes (AE1, AE2 and AE3) in the cholangiocytes and the hepatocytes, and also measure AE2 gene expression level after bile duct ligation (BDL). As we previously described, isolated hepatocytes and cholangiocytes from the liver of normal and BDL rats were prepared and gene expression levels were measured by using RT-PCR. We found that AE1, AE2, and AE3 genes were expressed in both hepatocytes and cholangiocytes, but CFTR was only in cholangiocytes. AE2 gene expression level was higher in the BDL hepatocytes than normal hepatocytes, which was significantly different between two groups. AE2 gene expression level was lower in the BDL cholangiocytes than normal cholangiocytes. However, AE2 gene expression level in both hepatocytes and cholangiocytes were not changed with a longer duration of BDL. These results suggest that CFTR and AE2 may play an important role in the pathogenetic mechanism of biliary cholestatic liver disease.

Probiotics have many effects such as antihypertensive, prevention of cancer, antioxidation, reduction of dermatitis symptoms, improvement of mineral absorption, reduction of allergic symptoms, and decrease of cholesterol, However, the main role of probiotics is that they balance intestinal microbials proportion. L. acidophilus is one of probiotics and microflora in intestine. It has an acidification activ-ity, aroma production, texture formation and probiotics properties. We studied on the roles of L. acid-ophilus in mice. In this study, body weights of mice were decreased when administration of L. acid-ophilus (1×10 10 CFU) and swimming ability has been raised than a normal group after feeding on L. acidophilus (1×10 10 CFU). After taking L. acidophilus (1×10 10 CFU), total white cells were increased than a normal group; hemoglobin and thrombocytes were increased. The level of cholesterol and tri-glyceride were decreased in blood analysis. We knew L. acidophilus is related to innate immune system. We found out the secretion of cationic peptide was increased in the Lysoplate assays as a re-sult of

Cordycepin (3'-deoxyadenosin), a specific polyadenylation inhibitor, is the main functional component in Cordyceps militaris, one of the top three renowned traditional Chinese medicines. Cordycepin has been shown to possess many pharmacological activities including immunological stimulation, and anti-bacterial, anti-viral, and anti-tumor effects. However, the mechanisms underlying its anti-cancer mechanisms are not yet understood. In this study, the apoptotic effects of cordycepin were investigated in human leukemia cells. Treatment with cordycepin significantly inhibited cell growth in a concentration-dependent manner by inducing apoptosis but not necrosis. This induction was associated with generation of reactive oxygen species (ROS), mitochondrial dysfunction, activation of caspases, and cleavage of poly(ADP-ribose) polymerase protein. However, apoptosis induced by cordycepin was attenuated by caspase inhibitors, indicating an important role for caspases in cordycepin responses. Administration of N-acetyl-l-cysteine, a scavenger of ROS, also significantly inhibited cordycepin-induced apoptosis and activation of caspases. These results support a mechanism whereby cordycepin induces apoptosis of human leukemia cells through a signaling cascade involving a ROS-mediated caspase pathway.

Human neuroblastoma is the most commonly diagnosed solid tumor in children. The existence and development of S-type cells is important for the prognosis and malignant properties in neuroblastoma patients. However, their origin is controversial and the relationship between S- and N-type cells in neuroblastoma has not yet been clarified. To investigate the feasibility of inter-conversion and characteristic features between S-type cells lacking malignant potential and N-type cells having metastatic potential, the genomic profiles of neuroblastoma SH-EP (S-type) and SH-SY5Y (N-type) cells were compared at high resolution. Common gain segments (>10 Mb) between SH-EP and SH-SY5Y cells were observed at 1q21.1–q44 and 17q21.32–q25.3. The results of the fluorescent in situ hybridization (FISH) analysis showed good agreement with the array-CGH data. Genome-wide inspection of SH-EP and SH-SY5Y cells successfully identified not only common chromosomal aberrations but also genomic variations, suggesting that interconversion could not take place between S- and Ntype cells. The identified differences between less aggressive S-type cells (SH-EP) and highly aggressive neuroblastic N-type cells (SH-SY5Y) might be useful for understanding tumorigenicity and discovery of potential new markers in neuroblastoma. This is the first effort to compare genomic profiles between less aggressive S-type cells (SH-EP) and highly aggressive neuroblastic N-type cells (SH-SY5Y) at high resolution.

The present study aims to evaluate Cordyceps militaris water extract (CMWE) with a view to develop melanogenesis inhibitors. Inhibitory activities of CMWE against tyrosinase, L-DOPA(L-3,4-dihydroxyphenylalanine) oxidation, and melanin biosynthesis in B16 mouse melanoma cells were investigated. CMWE, at 5000 μg/ml, inhibited tyrosinase activity of 71% and DOPA oxidation of 40% as reacting with L-DOPA. Furthermore, B16 mouse melanoma cell survived over 50% from low to high dose on MTT assay, and CMWE markedly inhibited (>50%) melanin synthesis at 5000 μg/ml. The inhibitory effect of CMWE on melanogenesis was attributed to enhancement of tyrosinase degradation. Key enzyme of melanin biosynthesis is tyrosinase which catalyses a beginning step from tyrosine to DOPA quinine and melanin formation step, respectively. These results indicated that CMWE may be a potential source of novel whitening agents for cosmetic or therapeutic application.

Ethnopharmacological relevance : Lonicera japonica Thunb and Anemarrhena asphodeloides BUNGE have been used for the treatment of a variety of inflammatory diseases, cold and infective diseases in many countries, including Korea and China. Aim of the study : This study aimed to assess the anti-nociceptive and anti-inflammatory activities of n-butanol fraction (WIN-34B) prepared from dried flowers of Lonicera japonica and dried roots of Anemarrhena asphodeloides as potential novel treatment of osteoarthritis. Materials and methods : Anti-nociceptive activities of WIN-34B (100, 200 and 400 mg/kg, p.o.) were measured using acetic acid-induced writhing response, formalin-induced paw licking, hot plate, radiant heat tail-flick, carrageenan-induced paw pressure, and Hargreaves tests, respectively. Anti-inflammatory activities of WIN-34B (100, 200 and 400 mg/kg, p.o.) were assessed using acetic acid-induced vascular permeability, carrageenan-induced paw edema, and croton oil-induced ear edema. Anti-osteoarthritis effect of WIN-34B was analyzed using monosodium iodoacetate (MIA)-induced osteoarthritis animal model. Results : WIN-34B exhibited better anti-inflammatory activity than that of celecoxib in carrageenan at the dose of 200 mg/kg and croton oil-induced paw edema and ear edema at the doses of 200 and 400 mg/kg. WIN-34B exhibited significant anti-inflammatory effects on vascular permeability. WIN-34B also exhibited significant anti-nociceptive activities in the late phase of formalin-induced paw licking and writhing response model in mice. In radiant heat tail-flick and carrageenan-induced paw pressure tests, WIN-34B at the dose of 400 mg/kg and at the doses of 200 and 400 mg/kg presented similar activities to indomethacin and celecoxib. Compared to indomethacin WIN-34B at 400 mg/kg showed similar or better anti-nociceptive activities after 1 and 2 h of theraphy in the hot plate test and better anti-nociceptive activity than that of celecoxib in Hargreves test. In the MIA-induced osteoarthritis animal models, WIN-34B at 400 mg/kg exhibited similar or better anti-nociceptive property than that of celecoxib throughout the pain measurement periods. Conclusion : When compared to celecoxib, WIN-34B exhibited similar or better anti-nociceptive and anti-inflammatory activities in osteoarthritic animal models, which may become a potential novel treatment for osteoarthritis.

Bee venom (BV) treatment is the therapeutic application of honeybee venom (HBV) for treating various diseases in Oriental medicine. In the present work, the authors investigated the functional specificity of BV as an angiogenesis inhibitor using in vitro models and in vivo mouse angiogenesis and lung metastasis models. BV significantly inhibited the viability of Lewis lung carcinoma (LLC) cells but did not affect peripheral blood mononuclear lymphocytes (PBML) cells. BV also inhibited vascular endothelial growth factor (VEGF)-induced proliferation, migration and capillary-like tube formation of human umbilical vein endothelial cells (HUVECs). Western blotting analysis showed that BV inhibited AKT and MAPK phosphorylation in LLC cells and HUVECs and down regulated expression of VEGF and VEGFR-2 of LLC cells and HUVECs. Also, BV effectively disrupted VEGF-induced neovascularization in Matrigel plugs in our in vivo angiogenesis assay. When given subcutaneously, BV also significantly suppressed tumor angiogenesis through inhibition of VEGF and VEGFR-2 in LLC model. Mice bearing subcutaneous LLC tumors were treated with 1mug/ml or 10mug/ml of BV. They showed reductions ranging between 49% and 62% in primary tumor volume and reduction of spontaneous pulmonary metastasis occurrences. Furthermore, BV treatment in the spontaneous lung metastases model after primary tumor excision prolonged their median survival time from 27 to 58days. These results suggest that the tumor-specific anti-angiogenic activity of BV takes effect during different stages of tumor progression by blocking the tyrosine phosphorylation of VEGFR-2, and validate the application of BV in lung cancer treatment.

The concentration of phenolics in Phellinus pini (CY001) extracts, expressed as mg of GAEs per g of P. pini fractions, and the EtOAc fraction (436.5 ㎎ GAEs/g) of P. pini had a higher phenolic content than other fractions. Several biochemical assays were used to screen antioxidant properties such as reducing power, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, NBT/XO superoxide system and inhibition of DCF/AAPH peroxyl radicals. Among the six mushroom extracts, the EtOAc fraction from P. pini (CY001) showed the most potent DPPH radical, superoxide radical, and peroxyl radical scavenging activities, with IC 50 values of 11.49 ㎍/ml, 8.32 ㎍/㎖, and 1.91 μg/㎖, respectively. The EtOAc fraction of P. pini (CY001) significantly inhibited enzymatic lipid peroxidation and effectively attenuated LPS-induced NO production of RAW 264.7 cells without cytotoxicity. We also found that the EtOAc fraction had a significant hepato-protectant effect on tacrine-induced cytotoxicity in HepG2 cells. These findings suggest that P. pini (CY001) may have potential as a natural antioxidant, which contains compound(s) with radical scavenging activity.

The aim of this study was to determine the in vitro anti-inflammatory effect of hot water extract from Cordyceps militaris fruiting bodies (CMWE) on lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) release in RAW 264.7 cells. The treatment of macrophages with various concentrations of hot CMWE significantly reduced LPS-induced production as well as NO, TNF-α and IL-6 secretion in a concentration-dependent manner. These results suggest that CMWE have potent inhibitory effects on the production of these inflammatory mediators.

Because tumor necrosis factor-alpha (TNF-alpha) is well-known to induce inflammatory responses, thus its clinical use is limited in cancer treatment. Rosmarinic acid (RA), a naturally occurring polyphenol flavonoid, has been reported to inhibit TNF-alpha-induced NF-kappaB activation in human dermal fibroblasts. However, the precise mechanisms of RA have not been well elucidated in TNF-alpha-mediated anti-cancer therapy. In this study, we found that RA treatment significantly sensitizes TNF-alpha-induced apoptosis in human leukemia U937 cells through the suppression of nuclear transcription factor-kappaB (NF-kappaB) and reactive oxygen species (ROS). Activation of caspases in response to TNF-alpha was markedly increased by RA treatment. However, pretreatment with the caspase-3 inhibitor, z-DEVD-fmk, was capable of significantly restoring cell viability in response to combined treatment. RA also suppressed NF-kappaB activation through inhibition of phosphorylation and degradation of IkappaBalpha, and nuclear translocation of p50 and p65. This inhibition was correlated with suppression of NF-kappaB-dependent anti-apoptotic proteins (IAP-1, IAP-2, and XIAP). RA treatment also normalized TNF-alpha-induced ROS generation. Additionally, ectopic Bcl-2 expressing U937 reversed combined treatment-induced cell death, cytochrome c release into cytosol, and collapse of mitochondrial potential. These results demonstrated that RA inhibits TNF-alpha-induced ROS generation and NF-kappaB activation, and enhances TNF-alpha-induced apoptosis.

The objective of this study was to determine the anticancer effects of the Pacific oyster (Crassostrea gigas) in vitro. For this study, the lipid extracts of C. gigas were prepared using several organic solvents: methanol, chloroform, hexane, methanol:chloroform=1:1, chloroform: hexane=1:1. The anticancer activity of the extracts was evaluated using cell cycle and apoptosis assays analyzed by a flow cytometry. Of all the extracts, the hexane extracts exhibited the highest anticancer activity compared to the other extracts. The hexane extracts were further separated and purified using thin layer chromatography. The final isolated lipid compounds were identified their components as palmitic acid, margaric acid, and stearic acid. These results indicate that this combination of lipids effectively inhibit in vitro human prostate tumor growth by inducing apoptosis of cancer cells.

Telomerase, a ribonucleoprotein that plays an important role in neoplastic immortality, is up-regulated in approximately 85% of cancers, especially in leukemia. The polyphenol, butein, has potent effects against various types of cancer cells, but its effects on telomerase activity have not been well characterized. In this study, we show that butein causes a down-regulation of hTERT gene expression and a concomitant decrease of telomerase activity. Butein also suppresses expression of c-Myc at the transcriptional level and down-regulates DNA-binding activity, regardless of cell type specificity, in leukemia cells. DNA-binding activities of c-Myc to the hTERT core promoter were decreased in butein-treated cells, as seen by chromatin immunoprecipitation assay. Treatment with butein also suppressed the activation of Akt, thereby inhibiting hTERT phosphorylation and translocation into the nucleus. In this process, butein also up-regulated the surface expression of CD11b in leukemia cells. Inhibition of telomerase activity by butein was followed by loss of proliferative capacity, induction of apoptosis, and differentiation. These findings demonstrate the effectiveness of butein at inhibiting telomerase activity by down-regulating hTERT gene expression in human leukemia cells.

The C. militaris belongs to entomopathogenic fungi, which have their specific sequences in internal transcribed spacer regions (ITS1 and ITS2) depending on species. In this study, to identify the phylogenetic relationship of the strain hybrided by mating of C. militaris, we compared DNA sequences of ITS regions and 5.8S ribosomal DNA (rDNA) repeat unit of hybrid strain and its parental strains. The result revealed that hybrid strains are C. militaris species. In addition, cordycepins produced by hybrid strains and other strains of C. militaris were analyzed by HPLC with 20mM of mobile phase and C-18 columns. The result indicated that the strain hybrided by mating produce higher concentration of phytochemical cordycepin than other C. militaris strains.

Anatomical features of both leaves and stems of the four mistletoes in Korea (Viscum album var. coloratum, Korthalsella japonica, Loranthus yadoriki, L. tanaka) and of their secondary haustorial structure within several host plants were investigated. Among the four mistletoes, there were diagnostic characters of the anatomy of leaves and stems which enabled us to distinguish the four taxa. Leaves were observed to have three distinct characters including unifacial or bifacial leaves, the number of vascular bundles in the midveins, and the level of development of sclerenchyma cells. There were four diagnostic characters of stems: overall morphology of stems in transverse view, degree of cuticle development, arrangement of vascular bundles, and features of the sclerenchyma and pith. In order to determine secondary haustorial traits, the research focused on the seven host plants of L. yadoriki and on the five host plants of K. japonica. The following features were found to be important: presence or absence of an aerial runner root, the shape of the haustorial strand and flange, the degree of penetration into host tissues, and their development of shaft in transverse view, the development both of secondary haustorial cells and short tracheid in hyphae. Korthalsella japonica and L. yadorki were clearly distinguished by these characters. The secondary haustorial forms in each host were somewhat different, due to varying degrees of development in the strength of the host plants' wood. However, qualitative characters like the final position of the secondary haustorial penetration into host tissues and the development of short tracheid cells were not only affected by the degree of development of the host plants, but also useful for the systematic study.

KHBJ-9B has been formulated by n-butanol fraction from 2 herbs known to have cartilage protection and anti-inflammatory effects. We elected to determine the osteoarthritic efficacy and mechanism of KHBJ-9B on human osteoarthritis cartilage explants culture and in a rabbit model of collagenase-induced osteoarthritis (CIA). The major chemical composition and quantification of KHBJ-9B was determined by high performance liquid chromatography. The efficacy of KHBJ-9B and its major compounds on cartilage protective effects such as inhibition of GAG release and type II collagen degradation, and their cytotoxicity in IL-1beta-treated human cartilage culture were examined. The mechanism of action of KHBJ-9B and its major compounds were evaluated by measuring inflammatory cytokines (IL-1beta and TNF-alpha) and matrix proteinases (ADAMTS-4, ADAMTS-5, MMP-1, MMP-13 and TIMP-3) in IL-1beta-treated human cartilage cultures. Also, the therapeutic effect of KHBJ-9B was confirmed using a collagenase-induced osteoarthritis (CIA) rabbit model. KHBJ-9B and 3 combined triterpenoids potently inhibited the release of proteoglycan and type II collagen in a dose dependent manner without cytotoxicity in IL-1beta-treated human cartilage explants culture, whereas its single major compounds (betulin, pimaradienoic acid and betulinic acid) and COX-2 inhibitor (NS398) showed little inhibition even at high concentrations. KHBJ-9B and the combination of 3 triterpenoids markedly inhibited the level of IL-1beta and TNF-alpha, and down-regulated the level of aggrecanases, ADAMTS-4, ADAMTS-5, MMP-1 and MMP-13, and up-regulated TIMP-3 in human cartilage explants culture. However, standard compounds and NS398 do not much affect the level of TNF-alpha, aggrecanases, and TIMP-3 in cartilage explants culture. In in vivo studies, KHBJ-9B significantly suppressed the stiffness level and global histologic score. Cartilage loss was significantly inhibited in the knee joint in a dose dependent manner, and this was associated with the finding that loss of proteoglycan, degradation of aggrecan and type II collagen was markedly reduced. These results suggest that the effect of KHBJ-9B is bigger than the effects of its single major compounds of triterpenoids or celecoxib inhibitors on cartilage protection and anti-inflammation in human cartilage and in in vivo model of osteoarthritis, and thus has potential for use in osteoarthritis treatment.

In this study, we found that pectenotoxin-2 (PTX-2) decreased cell viability and inhibited telomerase activity with downregulation of hTERT expression in human leukemia cells. PTX-2 treatment also reduced c-Myc and Sp1 gene expression and DNA binding activity. Further chromatin immunoprecipitation assay demonstrated that PTX-2 attenuated the binding of c-Myc and Sp1 to the regulatory regions of hTERT. We also observed that PTX-2 treatment attenuated the phosphorylation of Akt, thereby reducing the phosphorylation and nuclear translocation of hTERT. We concluded that PTX-2 suppressed telomerase activity through the transcriptional and post-translational suppression of hTERT and this process precedes cellular differentiation of human leukemia cells.Structured summaryMINT-6742762:hTERT (uniprotkb:O14746) physically interacts (MI:0218) with AKT (uniprotkb:P31749) by anti bait coimmunoprecipitation (MI:0006)

Siegesbeckia pubescens (S. pubescens) was widely used to alleviate symptoms of osteoarthritis (OA) in traditional medicine. However, the mechanism of action of S. pubescens remains unresolved. In the present study, we determined the physiological relevance of S. pubescens on cartilage protection in collagenase-induced osteoarthritis (CIA) in rabbits. The right knees of rabbits were injected intra-articularly with collagenase, and rabbits were orally administered with distilled water (vehicle), S. pubescens (100, 400 mg/kg) or celecoxib (100 mg/kg) once a day for 28 days after the initiation of the CIA. S. pubescens significantly suppressed the stiffness and global histological score including articular cartilage and synovial layer in CIA. Proteoglycan, aggrecan, and type II collagen expression was significantly increased in the rabbit knee joints of the S. pubescens–treated group. However, celecoxib had no effect on cartilage protection in CIA. The expression level of ADAMTS-4, ADAMTS-5, MMP-1, MMP-3, and MMP-13 were dose-dependently decreased in the S. pubescens–treated group. In contrast, the level of TIMP-1 dose-dependently increased. The pro-inflammatory cytokines involved in cartilage destruction, such as IL-1β, and inflammatory mediators containing PGE2 and NO were also inhibited in the S. pubescens–treated group. These results indicate that the cartilage protective effect of S. pubescens works through down-regulation of inflammatory mediators and aggrecanases and MMPs, while up-regulating TIMP-1 in the CIA rabbit model. Keywords:: Siegesbeckia pubescens, osteoarthritis, cartilage protection

Bee venom (BV), well known as a traditional Oriental medicine, has been shown to exhibit anti-arthritic and anti-carcinogenic effects. However, the molecular mechanisms responsible for the anti-inflammatory activity of BV have not been elucidated in microglia. In the present study, we investigated the anti-inflammatory effect of BV and its major component, melittin (MEL), on lipopolysaccharide (LPS)-stimulated BV2 microglia. Our results indicate that BV and MEL suppress LPS-induced nitric oxide (NO) and inducible NO synthase (iNOS) expression in a dose-dependent manner, without causing cytotoxicity in BV2 microglia. Moreover, BV and MEL suppressed LPS-induced activation of nuclear factor kappa B (NF-kappaB) by blocking degradation of IkappaBalpha and phosphorylation of c-Jun N-terminal kinase (JNK) and Akt, which resulted in inhibition of iNOS expression. Our data also indicate that BV and MEL exert anti-inflammatory effects by suppressing the transcription of cyclooxygenase (COX)-2 genes and proinflammatory cytokines, such as interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha. BV and MEL also attenuated the production of prostaglandin E(2) (PGE(2)). These results demonstrate that BV and MEL possess a potent suppressive effect on proinflammatory responses of BV2 microglia and suggest that these compounds may offer substantial therapeutic potential for treatment of neurodegenerative diseases that are accompanied by microglial activation.

The cytotoxic effect of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is limited in some cancer cells, including A549 lung adenocarcinoma cells. However, treatment with TRAIL in combination with subtoxic concentrations of sulforaphane (SFN) sensitizes TRAIL-resistant A549 cells to TRAIL-mediated apoptosis. Combined treatment with SFN and TRAIL induced chromatin condensation, DNA fragmentation, annexin V staining and sub-G(1) phase DNA content. These indicators of apoptosis correlate with the induction of caspase-3 activity that results in the cleavage of poly(ADP-ribose) polymerase and the release of lactate dehydrogenase. Both the cytotoxic effect and apoptotic characteristics induced by combined treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, demonstrating the important role of caspase-3 in the observed cytotoxic effect. Combined treatment also triggered the activation of p38 MAPK and JNK, and downregulation of ERK and Akt. Inhibitors of ERK (PD98059) or Akt (LY294002), but not p38 MAPK, resulted in significantly decreased cell viability. Although the activation of JNK was increased in response to combined treatment, inhibition of the JNK pathway significantly attenuated cell viability. These results indicate that caspase-3 is a key regulator of apoptosis in response to combined SFN and TRAIL in human lung adenocarcinoma A549 cells through downregulation of ERK and Akt.

Immunostimulating factor (ISTF) isolated from Actinobacillus actinomycetemcomitans which has been described previously, is distinct from lipopolysaccharide and induces proliferation of B cells. This study was undertaken to investigate whether ISTF might enhance the stimulation of other immune cells. Immunohistochemically, ISTF exhibited a profound stimulating effect on macrophages and dendritic cells as well as B cells in the spleen of BALB/c mice. ISTF was also recognized for its capacity to induce direct activation of mouse macrophages to produce IL-6, TNF-alpha, and NO and MHC class II expression. Therefore, it is postulated that ISTF stimulates macrophages and possibly other cells to produce a wide variety of proinflammatory mediators, which may be involved in the chronicity and tissue destruction of periodontal disease.

Although Fursarium oxysporum causes diseases in economically important plant hosts, identification of F. oxysporum formae speciales has been difficult due to confusing phenotypic classification systems. To resolve these complexity, we evaluated genetic relationship of nine formae speciales of F. oxysporum with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and translation elongation factor-1 alpha (EF-1α) gene. In addition, the correlation between mycotoxin content of fusaric acid and isolates based on molecular marker data was evaluated using the modified Mantel's test. According to these result, these fusaric acid-producing strains could not identify clearly, and independent of geographic locations and host specificities. However, in the identification of F. oxysporum formae speciales, especially, AFLP analysis showed a higher discriminatory power than that of a the RAPD and EF-1α analyses, all three techniques were able to detect genetic variability among F. oxysporum formae speciales in this study.