Echinatin induces reactive oxygen species-mediated apoptosis via JNK/p38 MAPK signaling pathway in colorectal cancer cells.

Colorectal cancer (CRC) is a very common and deadly cancer worldwide, and oxaliplatin is used as first-line chemotherapy. However, resistance usually develops, limiting treatment. Echinatin (Ech) is the main component of licorice and exhibits various therapeutic effects on inflammation-mediated diseases and cancer, ischemia/reperfusion, and liver injuries. The present study elucidated the underlying molecular mechanism of Ech-induced apoptosis in both oxaliplatin-sensitive (HT116 and HT29) and -resistant (HCT116-OxR and HT29-OxR) CRC cells. To evaluate the antiproliferative activities of Ech, we performed MTT and soft agar assays. Ech reduced viability, colony size, and numbers of CRC cells. The underlying molecular mechanisms were explored by various flow cytometry analyses. Ech-induced annexin-V stained cells, reactive oxygen species (ROS) generation, cell cycle arrest, JNK/p38 MAPK activation, endoplasmic reticulum (ER) stress, mitochondrial membrane potential depolarization, and multi-caspase activity. In addition apoptosis-, cell cycle-, and ER stress-related protein levels were confirmed by western blotting. Moreover, we verified ROS-mediated cell death by treatment with inhibitors such as N-acetyl-L-cysteine, SP600125, and SB203580. Taken together, Ech exhibits anticancer activity in oxaliplatin-sensitive and -resistant CRCs by inducing ROS-mediated apoptosis through the JNK/p38 MAPK signaling pathway. This is the first study to show that Ech has the potential to treat drug-resistant CRC, providing new directions for therapeutic strategies targeting drug-resistant CRC.

Isolinderalactone sensitizes oxaliplatin-resistance colorectal cancer cells through JNK/p38 MAPK signaling pathways.

BACKGROUND: Isolinderalactone (ILL), a sesquiterpene lactone compound, can be extracted from the root of Lindera aggregate. Physiological activities of ILL, including anti-inflammatory and anti-proliferative effects, have been investigated in multiple diseases. Nevertheless, little is known regarding its anti-cancer activities and the mechanism of action of ILL in targeting human CRC cells. PURPOSE: To determine ILL-mediated anti-proliferative effects on oxaliplatin (Ox)-sensitive and resistant colorectal cancer (CRC) cells and underlying mechanisms involved in its effects focusing on signal transduction. METHODS: Inhibitory effect of ILL on CRC cells was evaluated by analyzing mitochondrial membrane potential (MMP) dysfunction and multi-caspase activity. Apoptosis-regulating proteins and JNK/p38 signaling molecules were monitored by Western blotting. ROS-dependent physiological modifications by ILL were confirmed by pretreatment with N-acetylcysteine (NAC). Moreover, the involvement of JNK/p38 signaling in ROS-mediated apoptosis was verified by treatment with SP600125 (JNK inhibitor) and SB203580 (p38 inhibitor). RESULTS: ILL decreased cell viability and colony formation in both CRC Ox-sensitive (HCT116 and HT29) and Ox-resistant (OxR) (HCT116-OxR and HT29-OxR) cells. ILL induced G2/M phase cell cycle arrest, ROS generation, phosphorylated (p)JNK/p38 MAPK activation, mitochondrial membrane potential (MMP) depolarization, and multi-caspase activation, which eventually triggered apoptotic cell death of CRC cells. In addition, combined treatment with ILL and SP600125, SB203580, or pan-caspase inhibitor (Z-VAD-FMK) prevented decreases in cell viability seen after treatment with ILL alone. Pretreatment with NAC attenuated ILL-mediated apoptosis, ROS production, and p-JNK/p38 expression. CONCLUSION: Taken together, our results suggest that ILL can exert its anticancer effect in CRC Ox-sensitive and OxR cells by inducing ROS-mediated apoptosis through JNK/p38 MAPK signaling pathways. This is the first study demonstrating that ILL has a potential to improve drug efficacy against resistance mechanisms, providing a new insight into therapeutic strategies targeting drug-resistant CRC.

Therapeutic potential of α,ß-thujone through metabolic reprogramming and caspase-dependent apoptosis in ovarian cancer cells.

The therapeutic potential of α,ß-thujone, a functional compound found in many medicinal plants of the Cupressaceae, Asteraceae, and Lamiaceae families, has been demonstrated, including in inflammation and cancers. However, its pharmacological functions and mechanisms of action in ovarian cancer remain unclear. We investigated the anticancer properties of α,ß-thujone in ES2 and OV90 human ovarian cancer cells and its effect on sensitization to cisplatin. α,ß-thujone inhibited cancer cell proliferation and induced cell death through caspase-dependent intrinsic apoptotic pathways. Moreover, α,ß-thujone-mediated endoplasmic reticulum stress was associated with the loss of mitochondrial functions and altered metabolic landscape of ovarian cancer cells. α,ß-Thujone attenuated blood vessel formation in transgenic zebrafish, implying it has significant antiangiogenic potential. In addition, α,ß-thujone sensitized ovarian cancer cells to cisplatin, causing synergistic pharmacological effects. Collectively, our results suggest that α,ß-thujone has therapeutic potential in human ovarian cancer and functions via regulating multiple intracellular stress-associated metabolic reprogramming and caspase-dependent apoptotic pathways.

Lower Serum n-3 Fatty Acid Level in Older Adults with Sarcopenia.

The n-3 fatty acid (FA) has evoked considerable interest as a modifiable factor for maintenance of muscle health owing to its anti-inflammatory properties. To clarify this possibility, we investigated circulating n-3 FA level, a reliable biomarker of FA status in the body, in relation to sarcopenia in a cohort of Asian older adults. Blood samples were collected from 125 participants who underwent comprehensive assessment of muscle mass and function. Serum FA level was measured by gas chromatography/mass spectrometry. Sarcopenia was diagnosed using the cut-off points specified for the Asian population. After adjusting for sex, age, and body mass index, subjects with sarcopenia and those with low muscle strength had 36.5% and 32.4% lower serum n-3 levels (P = 0.040 and 0.030), respectively, than controls. The odds ratios per standard deviation increment in serum n-3 level for sarcopenia and low muscle strength were 0.29 and 0.40 (P = 0.015 and 0.028), respectively. A higher serum n-3 level was significantly associated with greater muscle strength (P = 0.038). These findings suggest a possible protective effect of n-3 FA on human muscle homeostasis. Further well-designed large-scale longitudinal studies are necessary to understand the definite role of circulating n-3 FA level in sarcopenia risk assessment.

Bavachin suppresses human placental choriocarcinoma cells by targeting electron transport chain complexes and mitochondrial dysfunction.

Phytoestrogens are naturally derived estrogen-like therapeutic compounds that have long been studied for their role as anti-cancer agents and supplements during chemotherapy. Bavachin is a therapeutic phytoestrogen used to treat cancer, inflammation, and diabetes mellitus. Though the therapeutic effects of bavachin on various diseases have been explored, its anti-cancer effects and related mechanisms in human placental choriocarcinoma remain unknown. This is the first study to identify the anti-cancer potential of bavachin on human placental choriocarcinoma cell lines JEG3 and JAR. Placental mitochondria support the elevated energy production required for placental development, through oxidative phosphorylation (OXPHOS). Based on this concept, we hypothesized that mitochondrial targeting by bavachin may contribute to anti-cancer activities in high-OXPHOS subtypes of cancer such as placental choriocarcinoma. Apoptosis and caspase activities were increased by bavachin in placental choriocarcinoma cells. Bavachin altered metabolic phenotypes by regulating electron transport chain complex and OXPHOS to suppress choriocarcinoma cell proliferation. It also led to calcium disruption and endoplasmic reticulum stress accompanied by mitochondrial membrane potential depolarization. It showed synergistic anti-cancer effects with paclitaxel on placental choriocarcinoma cells. Taken together, we suggest that bavachin has therapeutic potential against placental choriocarcinoma and may be used to counter paclitaxel-induced toxicity.

Beyond the fear: Nurses' experiences caring for patients with Middle East respiratory syndrome: A phenomenological study.

AIM AND OBJECTIVES: To explore the experiences of Korean nurses who had directly cared for patients with Middle East respiratory syndrome (MERS) and to derive the structure and meaning of these experiences. BACKGROUND: In 2015, the MERS epidemic struck Korea, and ill-prepared nurses had to care for patients with MERS. Nurses experienced conflict between their fear of the disease and their work and professional ethic. DESIGN: We employed a phenomenological qualitative approach. METHODS: Inductive, qualitative, in-depth interviews were performed with 17 nurses. The study process followed the Consolidated Criteria for Reporting Qualitative Research (COREQ) checklist. RESULTS: The qualitative inductive content analysis generated seven theme clusters and 18 themes. The theme clusters were "Fear of Uncertainty," "Beyond Hesitation," "A Scene Like a Battlefield," "Chaotic Nursing Identity," "Buttresses for Sustainability," "Lingering Trauma" and "Expanded Horizon of Nursing." The final analysis revealed that the core theme was "Beyond the fear of uncertainty." CONCLUSIONS: This study contrives a more in-depth, holistic understanding by describing the experiences of nurses who directly cared for patients with MERS-the first large-scale infectious disease in Korea. Although nurses saw themselves as vital caregivers, they were frightened of the disease, had to work in a harsh environment, experienced various internal conflicts and had to deal with varying forms of uncertainty. RELEVANCE TO CLINICAL PRACTICE: This study sheds light on the nursing situation during crises involving serious infectious diseases; to combat these, more medical facilities are needed, and staff should be proactively guided on how to care for patients. It can serve as part of a good foundation for further study of medical staff during recurring epidemics.

α,ß-Thujone suppresses human placental choriocarcinoma cells via metabolic disruption.

α,ß-Thujone is a natural terpenoid found in many medicinal herbs, such as Artemisia absinthium (wormwood), that exhibits antioxidant, anti-diabetic, and anti-tumorigenic effects. α,ß-Thujone has numerous functions; it serves as a food ingredient, cosmetic additive, and medicinal remedy. Although the therapeutic properties of α,ß-thujone were previously revealed, a comprehensive description of the mechanisms of its anti-cancer potential in choriocarcinoma is yet to be provided. To our knowledge, this study is the first to demonstrate that α,ß-thujone attenuates JEG3 and JAR choriocarcinoma cells through a caspase-dependent intrinsic apoptotic pathway. Moreover, α,ß-thujone was demonstrated to induce a global mitochondrial defect and ER stress in choriocarcinoma by causing mitochondrial depolarization, calcium overload, and metabolic alterations, thereby leading to energy deprivation, which eventually contributes to the increase in apoptosis of choriocarcinoma cells. Herein, we also revealed the synergistic anti-cancer activity of α,ß-thujone via its sensitization effect on paclitaxel in choriocarcinoma cells. Altogether, our findings suggest that α,ß-thujone is a novel, natural pharmacological compound that can be used to treat human placental choriocarcinoma.

Effect of ß-caryophyllene from Cloves Extract on Helicobacter pylori Eradication in Mouse Model.

New antibacterial treatments against Helicobacter pylori are needed as H. pylori is acquiring antibiotic resistance. ß-caryophyllene is a natural bicyclic sesquiterpene, with anti-inflammatory and antimicrobial effects. This study investigates the effects of H-002119-00-001 from ß-caryophyllene on the eradication of H. pylori in a mouse model, and its effects on the inflammation of the gastric mucosa. To evaluate the anti-H.pylori efficacy of ß-caryophyllene, a total of 160 mice were divided into eight groups (n = 10 each) and were administered different treatments for 2 and 4 weeks. H. pylori eradication was assessed using a Campylobacter-like organism (CLO) test and H. pylori qPCR of the gastric mucosa. The levels of inflammation of gastric mucosa were assessed using histology and immunostaining. H-002119-00-001 decreased bacterial burden in vitro. When H-002119-00-001 was administered to mice once daily for 2 weeks, cure rates shown by the CLO test were 40.0%, 60.0%, and 70.0% in groups 6, 7, and 8, respectively. H. pylori levels in gastric mucosa decreased dose-dependently after H-002119-00-001 treatment. H-002119-00-001 also reduced levels of inflammation in gastric mucosa. H-002119-00-001 improved inflammation and decreased bacterial burden in H. pylori-infected mouse models. H-002119-00-001 is a promising and effective therapeutic agent for the treatment of H. pylori infection.

Laminarin-Derived from Brown Algae Suppresses the Growth of Ovarian Cancer Cells via Mitochondrial Dysfunction and ER Stress.

Ovarian cancer (OC) is difficult to diagnose at an early stage and leads to the high mortality rate reported in the United States. Standard treatment for OC includes maximal cytoreductive surgery followed by platinum-based chemotherapy. However, relapse due to chemoresistance is common in advanced OC patients. Therefore, it is necessary to develop new anticancer drugs to suppress OC progression. Recently, the anticancer effects of laminarin, a beta-1,3-glucan derived from brown algae, have been reported in hepatocellular carcinoma, colon cancer, leukemia, and melanoma. However, its effects in OC are not reported. We confirmed that laminarin decreases cell growth and cell cycle progression of OC cells through the regulation of intracellular signaling. Moreover, laminarin induced cell death through DNA fragmentation, reactive oxygen species generation, induction of apoptotic signals and endoplasmic reticulum (ER) stress, regulation of calcium levels, and alteration of the ER-mitochondria axis. Laminarin was not cytotoxic in a zebrafish model, while in a zebrafish xenograft model, it inhibited OC cell growth. These results suggest that laminarin may be successfully used as a novel OC suppressor.

Environmentally friendly approach to recover vanadium and tungsten from spent SCR catalyst leach liquors using Aliquat 336.

This research paper deals with an environmentally friendly approach for the treatment of spent selective catalytic reduction (SCR) catalyst. To recover vanadium (V) and tungsten (W) from spent SCR catalyst, leach liquors from hydrometallurgical processing were utilized to develop a proper methodology for extraction and possible separation of vanadium and tungsten from each other. This study investigated the solvent extraction (also called liquid-liquid extraction) of vanadium and tungsten utilizing the alkaline roasted leached solution containing approximately ∼7 g L-1 of tungsten and ∼0.7 g L-1 of vanadium. The commercial extractant, N-methyl-N,N,N-tri-octyl-ammonium chloride [R3NCH3]+Cl- (commercial name Aliquat 336), was dissolved in Exxsol™ D80 (diluent) system and adopted in this research. Solvent extraction studies were performed to determine the following experimental parameters: equilibrium pH, extractant concentration, diluent influence, chloride ion concentration, temperature, and stripping reagent concentration, which were systematically scanned to ascertain the optimum conditions for quantitative extraction of both title metals. An anion exchange mechanism was proposed using the quaternary ammonium chloride solvent reagent after slope analysis. Excess supplement of chloride proved to have adverse effects, further supporting the extraction mechanism. Thermodynamics results show positive values for enthalpy (ΔH) for vanadium and tungsten, favoring the endothermic nature of the extraction reaction towards the uptake of either metal. McCabe-Thiele plots for extraction were constructed, suggesting 2 and 3 stages for vanadium and tungsten extraction, respectively, at the aqueous (A) to organic (O) phase ratio of 7 : 1, ensuring more than 99.9% and 7-fold enrichment of both title metals. The stripping trend follows the order: (NaOH + NaCl) > (NaOH + NaNO3) > NaOH > NaNO3 > NaCl. Stripping isotherm followed by stripping counter-current (CCS) study was carried out for quantitative stripping of the metals.

Phenolic compounds and antioxidant activity in sweet potato after heat treatment.

BACKGROUND: The ability of heat treatment with a soaking solvent to increase soluble phenolic compounds due to the liberation or breakdown of the cell matrix has been investigated in various plants. This study investigated the changes in phenolic compounds and antioxidant activities of 12 sweet potato cultivars after heat treatment with distilled water or prethanol A. RESULTS: The highest total polyphenol content (134.67 mg gallic acid equivalents/g extract residue) and flavonoid content (65.43 mg catechin equivalents/g extract residue) was observed in the 'Jami' (JM) cultivar after heat treatment with prethanol A. Higher polyphenol and flavonoid content was generally observed in the purple sweet potato cultivars. Salicylic acid was the major phenolic acid, followed by protocatechuic acid or chlorogenic acid in almost all untreated sweet potato cultivars. The salicylic acid, vanillic acid, gallic acid, and caffeic acid content of the sweet potatoes increased after the heat treatment, whereas the protocatechuic acid and chlorogenic acid content decreased. The highest 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azinobis(3-ethyl benzothiazoline)-6-sulfonic acid (ABTS) radical scavenging activity levels were observed in the JM cultivar subjected to heat treatment with prethanol A (48.15 and 80.00 mg TE/g extract residue, respectively). CONCLUSION: These results suggest that heat treatment with a soaking solvent is an efficient method to enhance the antioxidant characteristics of Korean sweet potato cultivars. © 2019 Society of Chemical Industry.

Studies on Core-Shell Nanocapsules of Felodipine: In Vitro-In Vivo Evaluations.

The present study aimed for in vitro-in vivo-in silico simulation studies of experimentally designed (32-factorial) Capmul PG-8-cored, Eudragit RSPO-Lutrol F 127 nanocapsules to ferry felodipine using GastroPlus™. The in silico parameter sensitivity analysis for pharmacokinetic parameters was initially assessed to justify the preparation of felodipine-loaded nanocapsules (FLNs) with enhanced solubility to overcome the bioavailability issues of felodipine. The overall integrated desirability ranged between 0.8187 and 0.9488 for three optimized FLNs when analyzed for mean particle size, zeta potential, encapsulation efficiency, and in vitro dissolution parameters. The morphological evaluation (SEM, TEM, and AFM) demonstrated spherical nanoparticles (200-300 nm). Validated LC-MS/MS analysis demonstrated enhanced relative bioavailability (13.37-fold) of optimized FLN as compared to suspension. The simulated regional absorption of the FLN presented significant absorption from the cecum (26.3%) and ascending colon (20.1%) with overall absorption of 67.4% from the GIT tract. Furthermore, in vitro-in vivo correlation demonstrated the Wagner-Nelson method as the preferred model as compared to mechanistic and numerical deconvolution on the basis of least mean absolute prediction error, least standard error of prediction, least mean absolute error, and maximum correlation coefficient (r 2 = 0.920). The study demonstrated enhanced oral absorption of felodipine-loaded nanocapsules, and GastroPlus™ was found to be an efficient simulation tool for in vitro-in vivo-in silico simulations.

Celastrol blocks binding of lipopolysaccharides to a Toll-like receptor4/myeloid differentiation factor2 complex in a thiol-dependent manner.

ETHNOPHARMACOLOGICAL RELEVANCE: Tripterygium wilfordii (lei gong teng; Thunder of God Vine), which belongs to the Celastraceae family, has long been used in traditional Chinese medicine to treat inflammation and rheumatoid arthritis. Celastrol is a bioactive compound isolated from T. wilfordii. AIM OF THE STUDY: We investigated whether celastrol suppressed binding of lipopolysaccharides (LPS) to myeloid differentiation factor 2 (MD2), thereby downregulating Toll-like receptor4 (TLR4) activation in mouse primary macrophages. MATERIALS AND METHODS: Cytokine expression was determined by polymerase chain reaction analysis and enzyme-linked immunosorbent assay in bone marrow-derived primary macrophages (BMDMs). The kinase activity of tank-binding kinase 1 (TBK1) was examined by a luciferase reporter assay and an in vitro kinase assay. LPS binding to MD2 was examined by an in vitro binding assay and confocal microscopy analysis. RESULTS: Celastrol reduced LPS-induced expression of inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-12, and IL-1ß, at both the mRNA and protein levels in BMDMs. Celastrol suppressed LPS binding to MD2, as shown by the in vitro binding assay, whereas it did not inhibit TBK1. In addition, co-localization of LPS with MD2 in BMDMs was blocked by celastrol. The inhibitory effects of celastrol on LPS binding to MD2 were reversed by thiol donors (N-acetyl-L-cysteine and dithiothreitol), suggesting that the thiol reactivity of celastrol contributes to its inhibitory effects on TLR4 activation in macrophages. CONCLUSION: Our results demonstrate that celastrol suppresses TLR4 activation through the inhibition of LPS binding to the TLR4/MD2 complex. These results provide a novel mechanism of action by which celastrol contributes to the anti-inflammatory activity of T. wilfordii.

PLC-δ1-Lf, a novel N-terminal extended phospholipase C-δ1.

Phospholipase C-δ (PLC-δ), a key enzyme in phosphoinositide turnover, is involved in a variety of physiological functions. The widely expressed PLC-δ1 isoform is the best characterized and the most well understood phospholipase family member. However, the functional and molecular mechanisms of PLC-δ1 remain obscure. Here, we identified that the N-terminal region of mouse PLC-δ1 gene has two variants, a novel alternative splicing form, named as long form (mPLC-δ1-Lf) and the previously reported short form (mPLC-δ1-Sf), having exon 2 and exon 1, respectively, while both the gene variants share exons 3-16 for RNA transcription. Furthermore, the expression, identification and enzymatic characterization of the two types of PLC-δ1 genes were compared. Expression of mPLC-δ1-Lf was found to be tissue specific, whereas mPLC-δ1-Sf was widely distributed. The recombinant mPLC-δ1-Sf protein exhibited higher activity than recombinant mPLC-δ1-Lf protein. Although, the general catalytic and regulatory properties of mPLC-δ1-Lf are similar to those of PLC-δ1-Sf isozyme, the mPLC-δ1-Lf showed some distinct regulatory properties, such as tissue-specific expression and lipid binding specificity, particularly for phosphatidylserine.

Ginkgo biloba extract attenuates hyperalgesia in a rat model of vincristine-induced peripheral neuropathy.

BACKGROUND: Chemotherapy-induced peripheral neuropathy is a common, dose-limiting side effect of cancer chemotherapeutic drugs. Hyperalgesia is a common component of neuropathic pain. Ginkgo biloba extract (GBE) is an oriental herbal medicine that has various pharmacological actions. In this study, we evaluated the effects of oral GBE on hyperalgesia in a rat model of vincristine-induced neuropathy. METHODS: Male Sprague-Dawley rats (200-250 g) were injected intraperitoneally with vincristine or saline (0.1 mg/kg/d) using a 5-day-on, 2-day-off schedule over 12 days. All the behavioral tests for mechanical, cold, and heat hyperalgesia were conducted before the daily injection during the course of vincristine treatment. Rats that developed hyperalgesia 14 days after vincristine injection were randomly assigned into 4 groups. Distilled water and GBE (50, 100, and 150 mg/kg) were administered, respectively, to the individual groups. We examined the hyperalgesia at preadministration and at 15, 30, 60, 90, 120, 150, and 180 minutes after oral drug administration. RESULTS: Saline injection did not have any significant effect on mechanical, cold, and heat hyperalgesia. Vincristine injection produced mechanical and cold hyperalgesia. For the GBE groups, the paw withdrawal threshold to mechanical stimuli was significantly increased and withdrawal frequency to cold stimuli was significantly reduced versus the control group dose-dependently (P < 0.05). CONCLUSIONS: This study demonstrates that oral administration of GBE is associated with a dose-dependent antihyperalgesic effect on mechanical and cold stimuli in a rat model of vincristine-induced neuropathy.

Progression detection capability of macular thickness in advanced glaucomatous eyes.

PURPOSE: To evaluate the progression rate of macular and circumpapillary retinal nerve fiber layer (RNFL) thickness in advanced glaucomatous eyes using spectral domain optical coherence tomography (Cirrus HD-OCT, Carl Zeiss Meditec, Dublin, CA). DESIGN: Longitudinal, observational study. PARTICIPANTS: A total of 98 eyes of 98 patients with advanced glaucoma (visual field [VF] mean deviation [MD] <-10 dB) with a mean follow-up time of 2.2 years. METHODS: Three glaucoma experts independently reviewed optic disc and RNFL photographs and classified patients into 3 groups: progressed, stable, and undetermined (criterion 1). Patients in the undetermined group could not be evaluated because of advanced optic disc cupping. The eyes were also classified into 2 groups, progressed and stable, by serial VF data (criterion 2). MAIN OUTCOME MEASURES: Progression rates as determined by linear regression analysis against patient age using serial macular and RNFL thickness parameters were compared among different groups. RESULTS: By criterion 1, 25 eyes (25.5%) were classified as stable, 13 eyes (13.3%) were classified as progressed, and 60 eyes (61.2%) were classified as undetermined. By criterion 2, 86 eyes (87.8%) were classified as stable, and 12 eyes (12.2%) were classified as progressed. By criterion 1, the mean progression rate of average macular thickness was significantly higher in the progressed group than in the stable and undetermined groups (-4.74±4.40, -0.53±1.44, and -2.72±4.75 µm/year, respectively; P = 0.01). The undetermined group showed a higher progression rate than the stable group (P = 0.045). However, the progression rate of average RNFL thickness did not differ significantly among the 3 groups (-1.19±2.62, -0.33±1.29, and -1.21±2.75 µm/year, respectively; P = 0.34). By criterion 2, the mean progression rate of average RNFL thickness did not differ significantly between the stable and progressed groups (-0.90±2.42 and -2.08±2.85 µm/year; P = 0.459). However, the progression rate as revealed by average macular thickness was significantly different between the 2 groups (-2.22±4.33 and -5.12±2.40 µm/year, respectively; P = 0.039). CONCLUSIONS: Exploration of changes over time in macular thickness may improve detection of progression in patients with advanced glaucoma.

Production of cephalosporin C using crude glycerol in fed-batch culture of Acremonium chrysogenum M35.

In this study, cephalosporin C production by Acremonium chrysogenum M35 cultured with crude glycerol instead of rice oil and methionine was investigated. The addition of crude glycerol increased cephalosporin C production by 6-fold in shake-flask culture, and also the amount of cysteine. In fed-batch culture without methionine, crude glycerol resulted only in overall improvement in cephalosporin C production (about 700%). In addition, A. chrysogenum M35 became highly differentiated in fed-batch culture with crude glycerol, compared with the differentiation in batch culture. The results presented here suggest that crude glycerol can replace methionine and plant oil as cysteine and carbon sources during cephalosporin C production by A. chrysogenum M35.

Vascular smooth muscle dysfunction and remodeling induced by ginsenoside Rg3, a bioactive component of ginseng.

Ginseng, one of most well-known herbal medicines, is widely and indiscreetly used among the patients with cardiovascular disorders, raising concern over abuse of this medicine and unwanted effects. In this study, we investigated the effects of ginsenoside Rg3 (Rg3), an active ingredient of ginseng, on vascular contractility and structural integrity to explore its potential vascular toxicity. In isolated rat aorta, Rg3 suppressed the normal agonist-induced contractile response. This suppression persisted even after a rigorous washout. In the endothelium-denuded aortic ring, impairment of vascular contractility by Rg3 was retained, suggesting that vascular smooth muscle was affected. In primary vascular smooth muscle cells, Rg3 abolished agonist-induced Ca(2+) increase, indicating that Ca(2+) regulation was disrupted. Rg3 suppressed the contraction induced by Bay K8644, an L-type Ca(2+) channel activator, whereas store-operated Ca(2+) channel or intracellular Ca(2+) store-mediated contraction was not affected, suggesting that the L-type Ca(2+) channel was selectively impaired by Rg3. These in vitro results were further confirmed in vivo where Rg3 treatment significantly attenuated the agonist-induced pressor response. More importantly, 4-week repeated treatment with Rg3 in normal animals induced eutrophic outward remodeling in the thoracic aorta, that is, it brought about an increased luminal area without changes in the wall area. These results suggest that Rg3 can induce the vascular smooth muscle dysfunction by disturbing Ca(2+) influx from the L-type Ca(2+) channel, ultimately leading to impaired vascular contractility and structural remodeling.

Natural ortho-dihydroxyisoflavone derivatives from aged Korean fermented soybean paste as potent tyrosinase and melanin formation inhibitors.

Natural o-dihydroxyisoflavone (ODI) derivatives with variable hydroxyl substituent at the aromatic ring of isoflavone and three known isoflavones were isolated from five-year-old Korean fermented soybean paste (Doenjang) and evaluated as potent inhibitors on tyrosinase activity and melanin formation in melan-a cells comparing with other known isoflavones, 7,8,4'-trihydroxyisoflavone (1) and 7,3',4'-trihydroxyisoflavone (2) inhibited tyrosinase by 50% at a concentration of 11.21+/-0.8 microM and 5.23+/-0.6 microM (IC(50)), respectively, whereas, 6,7,4'-trihydroxyisoflavone (3), daidzein (4), glycitein (5) and genistein (6) showed very low inhibition activity. Furthermore, those compounds significantly suppressed the cellular melanin formation by 50% at a concentration of 12.23+/-0.7 microM (1), 7.83+/-0.7 microM (2), and 57.83+/-0.5(6) and show more activity than arbutin. But, compounds 3, 4, and 5 showed lower inhibition activity. This study shows that the position of hydroxyl substituent at the aromatic ring of isoflavone plays an important role in the intracellular regulation of melanin formation in cell-based assay system.

U-shaped dose response in vasomotor tone: a mixed result of heterogenic response of multiple cells to xenobiotics.

U-shaped response has been frequently encountered in various biological areas including epidemiology, toxicology, and oncology. Despite its frequent observation, the theory of U-shaped response has been crippled by the lack of a robust mechanism underlying and incomplete in vitro and in vivo correlation. In the present study, a novel mechanism is provided for a U-shaped response, based on the findings of agonist-induced vasomotor tone change affected by menadione (MEN) (synthetic vitamin K(3)), a reactive oxygen species generator, and arsenic, an environmental pollutant, which showed typical U-shaped responses in both in vitro aortic contractile response and in vivo blood pressure. U-shaped responses by MEN and arsenic were a combined result from heterogenic susceptibilities and responses of multiple target cells composing blood vessels, that is, endothelium and smooth muscle. Notably, endothelium, a regulator of vasomotor tone, was primarily affected by low-dose stimuli, whereas smooth muscle, an effector of vascular contraction, was affected later by high-dose. The dysfunction of smooth muscle was produced by high-dose MEN-induced hydrogen peroxide, resulting in the attenuation of vascular contractile reactivity, whereas low-dose MEN-induced superoxide led to the quenching of vasodilatory nitric oxide in endothelial cells, resulting in the enhancement of vasoconstriction. This mechanistic theory, the difference in susceptibilities and responses to a common stimulus between regulator and effector components of a system, could give a new insight into the explanation of various U-shaped responses and provide a new evidence for the need of the risk assessment of toxicants with a wider dose range.