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A continuous purification process can be beneficial to the purification of biologics due to its higher productivity and efficiency than a conventional batch purification process. However, regulatory issues and lack of established cases render deployment of the continuous process difficult in industrial settings. Here we report a case study for design and optimization of an advanced continuous process for purifying a low-titer enzyme as a model biologic. To convert a conventional batch process to an advanced continuous one in purification of biologics, conventional unit operations (UOs), including ultrafiltration/diafiltration (UF/DF) and batch chromatography, were replaced by advanced ones such as in-line dilution conditioning (IDC) and periodic counter-current chromatography (PCC). The UF/DF UO was changed to IDC UO to adjust pH and conductivity. The mixing ratio of the sample and the conditioning buffer in IDC was determined by experiments with three buffers. PCC was optimized with two variables, column height and sample loading residence time, as the delta pressure in the columns was less than 1.0 bar. A graph indicating the operating area was plotted to efficiently control the PCC. Although the sample volume increased in IDC, PCC had a complementary advantage in that purification was performed faster than batch chromatography. We observed at least 25% increase in economic advantage when the advanced continuous process was applied to purify a low-titer enzyme. We propose not only a continuous process with the substitution of UF/DF and batch chromatography with IDC and PCC but also a method to optimize PCC by plotting operating areas. [ABSTRACT FROM AUTHOR]

Oysters are rich nutrition sources that contain polysaccharides, proteins, peptides, phenolic compounds, minerals, and vitamins. In the search for compounds that exert anti-inflammatory effects in macrophages, we prepared hydrolyzed oyster (Magallana gigas) extracts (HOE) and evaluated their protective effects against inflammation and oxidative stress in RAW264.7 murine macrophages treated with lipopolysaccharide (LPS). As expected, LPS notably stimulated the generation of inflammatory molecules and associated regulatory proteins. However, HOE effectively blocked these effects in a concentration-dependent manner. This suppressive effect of HOE on inflammation appears to be responsible for blocking nuclear factor-κB (NF-κB) signaling. In addition, LPS-mediated reactive oxygen species generation RAW264.7 cells was highly inhibited by HOE treatment, which appears to be partially regulated by nuclear factor erythroid 2-related factor 2 (Nrf2). Moreover, HOE at high concentrations inhibited pro-inflammatory mediators and cytokines to a greater extent than BAY7085, a pharmacological NF-κB inhibitor, in LPS-treated RAW264.7 cells. Taken together, our results show that HOE effectively inhibits inflammation and oxidative stress via modulating the NF-κB and/or Nrf2 signaling in RAW264.7 macrophages and can be a potential therapeutic agent to prevent inflammation-related diseases. [ABSTRACT FROM AUTHOR]

The threats of air pollution mediated by particulate matter (PM) to human health are severe burdens to modern society. For example, PM is inhaled in the lungs or affects keratinocytes to induce reactive oxygen species that exert oxidative stress, inflammatory responses, and genotoxicity. Natural products are rich sources of the antioxidant and anti-inflammatory chemicals that might counteract what PM does in the human body. This review provides a compilation of knowledge on bioactive natural products that improve the health conditions from PM-induced health outcomes. The natural products were classified according to biosynthetic origins or resources: phenolic compounds including flavonoids and other phenolics, terpenoids, steroids, phytochemicals from marine algae, statins, lipids, alkaloids, and other nitrogen-containing natural products. This review article covers the structures and biological activity of natural products investigated with tissue cells or animal models. [ABSTRACT FROM AUTHOR]

Particulate matter 2.5 (PM2.5) is the biological toxic substances in the air pollutants, and causes to dry eye disease. To date, studies on the development of treatment for PM2.5-mediated dry eye disease are extremely limited. Schisandrae Fructus (SF), the fruit of Schisandra chinensis Turcz. (Baillon), has been used as traditionally herbal medicine in Northeastern Asia. SF has various pharmacological effects, but its effect on the ocular system have not been well defined. The aim of the present study was to investigate the effect of SF ethanol extract (SFE) on topical PM2.5-mediated dry eye symptoms in rats. SFE ameliorated tear deficiency, corneal epithelium detachment, reduction of conjunctival goblet cell, and inflammation of cornea and lacrimal gland induced by PM2.5 exposure. In addition, SFE markedly suppressed PM2.5-induced changes in the retinal composition, including loss of ganglion cells. Furthermore, oral administration of SFE significantly restrained PM2.5-induced increasing levels of total cholesterol and low low-density lipoprotein cholesterol. In conclusion, oral application of SFE may have protective effects against dry eye disease by PM2.5 exposure that result from restoration of tear film and inhibition of inflammation, and may in part contribute to improvement of retinal disorder and dyslipidemia. [ABSTRACT FROM AUTHOR]

Orthogonal aminoacyl tRNA synthetase (aaRS)/tRNA pairs are an efficient tool for the site-specific introduction of para-azido-L-phenylalanine (pAzF), a non-canonical amino acid, into the amber codon of proteins. In the present study, to improve amber suppression by aaRS, random mutagenesis using error-prone polymerase chain reaction method was carried out, and mutants with enhanced pAzF-incorporation ability were selected using fluorescence-activated cell sorting (FACS). For screening, the mutant superfolder green fluorescent protein with an amber codon was used as the reporter. Furthermore, the predicted structure of resulting aaRS mutants were analyzed. Three aaRS mutants — K776, K801, and M320 — showed 2.9-, 1.5-, and 3.7-times greater pAzF-incorporation ability, respectively, than did the control. The mutants also exhibited increased selectivity for pAzF introduction. The predicted protein models for the aaRS mutants showed that mutations in the anticodon and acceptor stem of the tRNA recognition region affected the tRNA binding affinity. These results demonstrate that random mutagenesis can cover the missed possibility of rational design or site directed mutagenesis, and screening with FACS is an efficient and rapid method for the evolution of aaRS. It can be also utilized for the development of other proteins with a variety of non-canonical amino acids. [ABSTRACT FROM AUTHOR]

Agarwood is a resinous wood from the tree in the family Thymelaeaceae. It has been used as traditional medicine in China and Southeast Asia countries. This study was conducted to evaluate the potential of agarwood pill (AP) as a supplement to enhance immune function. In this study, immunity-enhancing activity was demonstrated in BALB/c male mice administered with AP for 10 days after intraperitoneal injection of cyclophosphamide (CPA). Results demonstrated that AP supplementation ameliorated spleen index and body weight changes of CPA-injected mice by increasing the proliferation of T- and B-lymphocytes as well as the activity of natural killer cells. Analysis of spleen-derived lymphocytes showed that counts of CD4+ and CD8+ T-cells were slightly recovered in the spleen of mice treated with a high dosage of AP. Serum levels of immune-related cytokines of tumor necrosis factor-α were notably restored after AP administration. In addition, serum levels of immunoglobulin (Ig) M and IgG were recovered by AP administration at a medium dosage. Furthermore, in analysis of ex vivo bone marrow-derived macrophages and dendritic cells, counts of immune-related interleukin-12, major histocompatibility complex (MHC)-I, MHC-II, CD80, and CD86 were significantly increased by AP treatment in concentration-dependent manner. This study provides reference data for the development of AP as a supplementation to enhance human immune function. [ABSTRACT FROM AUTHOR]

This study was conducted to investigate the physicochemical properties and anti-inflammatory effects of the enzymatic extracts of Sargassum patens C. Agardh (SP) using crude enzyme from Shewanella oneidensis PKA 1008. SP is a brown alga containing 30–67% carbohydrates and polysaccharides, such as alginate, laminaran, and fucoidan. The contents of moisture, crude protein, crude lipid, ash, carbohydrate, and alginate in SP were 8.29, 18.10, 0.60, 21.64, 51.37, and 32.37%, respectively. The S. oneidensis PKA 1008 strain was isolated from Ulva pertusa and used to degrade alginate. To obtain the optimum degradation conditions, SP and crude enzyme from S. oneidensis PKA 1008 were prepared in a ratio of 1:1(v/v). The mixture was incubated at 30°C for 0–48 h. The changes in pH, color value, reducing sugar, viscosity, and TLC of SP-degrading extract were confirmed. The pH and viscosity significantly decreased at 24 h compared to those at 0 h. The highest reducing sugar concentration of the enzymatic extract was 571.91 µg/mL at 24 h. The enzymatic extracts were visualized by thin-layer chromatography (TLC) using a solvent system of 1-butanol: methanol: water, 4:1:2 (v/v) and degraded into monomers at 24 h. The anti-inflammatory effects of the enzymatic extracts at 0 and 48 h were measured using LPS-induced RAW 264.7 cells. These results indicate that crude enzyme from S. oneidensis PKA 1008 can be used to enhance the polysaccharide degradation of SP and its anti-inflammatory effect. [ABSTRACT FROM AUTHOR]

Tremella fuciformis TFC6 mycelia was isolated from its fruiting body. T. fuciformis TFCUV5 was obtained from TFC6 mycelia by ultraviolet mutagenesis to improve extracellular-polysaccharide (EPS) yield. In addition, we investigated the physicochemical properties and functionality of EPS for industrial applications and the effect of EPS on enhancement of the moisturizing factor and antiwrinkle effect in skin cells. The EPS yield was improved upon fermentation by the mutant strain through optimization of the medium composition and culture conditions to a maximum of 8.4 g/L; the productivity was 1.9 g/L-day under optimal culture conditions (initial pH 7.0 at 25°C, 300 rpm, 2 vvm). EPS with a molecular weight of 1.79 × 106 (± 0.721%) comprised mannose, xylose, glucuronic acid, fucose, and glucose. EPS extract increased both moisture and the antiwrinkle factor in skin cells. Therefore, EPS from TFCUV5 might have potential to be used in the cosmetics industry. [ABSTRACT FROM AUTHOR]

Mevastatin (HMG-CoA reductase inhibitor) isolated from Penicillium citrinum is known to the first statin that inhibitor of cholesterol synthesis. Recently, several clinical reports of other statins suggest its role on reducing the severity of psoriasis, but the available evidence on cellular/molecular level of mevastatin is unclear. Thus, this study was to determine the molecular/cellular mechanism of mevastatin using TNF-α/IL-17A stimulated HaCaT cells. First, the safety and effectiveness of mevastatin were confirmed through the comparative experiment of five statin derivatives. Mevastatin decreased mRNA levels of cytokines (IL-1α/β, IL-6, and IL-8), chemokine (CC motif) ligand 20 (CCL20), and keratin proteins (Keratin 5/14/6/16) known to be overexpressed in psoriasis. In addition, mevastatin significantly reduced phosphorylation of NF-κB, MAPKs and STAT3 in TNF-α/IL-17 signaling pathway. Moreover, mevastatin specifically suppressed keratin 5/14 proteins expressed in the keratinocyte differentiation process of the basal layer and keratin 6A/16 overexpressed in patients with psoriasis. These results imply that mevastatin not only can inhibit psoriasis-induced inflammatory cytokines and chemokines, but also can suppress psoriasis-related keratin proteins. [ABSTRACT FROM AUTHOR]

Microglial activation is critical for obesityinduced hypothalamic inflammation and is closely associated with pathologies of metabolic complications. In this study, we investigated the effect of filbertone, a main aroma compound of hazelnuts, on microglia-mediated inflammatory responses in vitro and obesity-induced hypothalamic inflammation in vivo. BV2 microglial cells were stimulated with lipopolysaccharide (LPS) in the presence or absence of filbertone. Meanwhile, C57BL/6 mice were fed for 10- weeks on a high-fat diet (HFD) supplemented with 0.2% filbertone. Levels of inflammatory mediators in microglia or hypothalamus were measured using enzyme-linked immunosorbent assays or quantitative real-time PCR. Filbertone significantly inhibited nitrite oxide production, inducible nitric oxide synthase expression, and inflammatory cytokine production in LPS-stimulated microglia. Filbertone also inhibited LPS-stimulated activation of inflammatory signaling molecules, mitogen-activated protein kinases (MAPK) such as extracellular signal-regulated kinases and p38, and the degradation of inhibitory nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in microglia. Moreover, filbertone supplementation markedly suppressed the expression of inflammatory cytokines and microglia activation marker in the hypothalamus of obese mice fed a HFD. These results suggest that filbertone reduces HFD-induced microglial activation through inhibition of the MAPK and NF-κB pathways, and thus protects obesityinduced hypothalamic inflammation. Filbertone may be useful for protection of microglia-mediated hypothalamic inflammation in obese condition and related metabolic complications. [ABSTRACT FROM AUTHOR]

A fibroin-degrading bacterium was isolated in a medium containing fibroin as a sole nitrogen source, and then identified and characterized. The strain was designated Bacillus subtilis strain Bs5C, and the culture medium containing the extracellular proteolytic enzymes was found to hydrolyze cocoon fiber. A decomposition rate of 35% was achieved with the culture medium in optimized cocoon-degrading condition. In a second round of decomposition performed on the same fibers with additional culture medium, the decomposition rate reached 45.9%. To investigate the proteolytic enzymes degrading cocoon, the enzymes was purified from culture medium using ion exchange column and identified using LC-MS/MS analysis system. As a result, it was uncovered that the cocoon degradation was due to the neutral protease NprE of the strain Bs5C. Moreover, the cocoon fibers treated with semi-purified NprE enzyme solution were clearly split and degraded, as assessed by SEM, and solubilized peptides from fibroin and sericin were detected by LC-MS/MS. In conclusion, this study is the first report that peptides could be produced from cocoon by cultured medium of B. subtilis, and the newly isolated strain Bs5C and the NprE protease from the strain Bs5C could clearly be valuable for the production of silk peptides, which have various pharmacological and industrial applications. [ABSTRACT FROM AUTHOR]

Stress is an emerging and inevitable entity. Stress causes anxiety, depression, imbalance of homeostasis, hormone imbalance, and sleep disturbances that are closely related to the onset of metabolic syndrome. The objective of this study was to investigate effects of Ocimum sanctum extract (TZ-OSE) on anti-anxiety, anti-stress, and control of physiological changes due to sleep disturbance in a rat model with restraint stress and sleep disorder induced. After inducing the rat model with restraint stress, three behavioral tests were performed and blood cortisol concentration was determined. In Elevated plus maze test, the test substance TZ-OSE at dose of 200 mg/kg showed the best effect in increasing time that stayed on the open arms. In tail suspension test (TST) and forced swimming test (FST), TZ-OSE significantly reduced immobility duration time (IMT). TZ-OSE also decreased plasma cortisol concentration. In the model after inducing sleep disturbance, ELISA, Western blotting, and Immuno-histochemistry (IHC) were conducted. TZ-OSE treatment decreased plasma cortisol, norepinephrine, glucose, and acylated ghrelin concentrations in the rat model with sleep disorders induced. Decreases of melatonin and leptin levels in the plasma of the rat model with sleep disorders induced were increased by TZ-OSE treatment. Finally, TZ-OSE treatment decreased metabolism-related markers (p-AMPK, Glut-4, p-ACC) in stomach and muscle tissues and decreased the expression of cortisol in adrenal gland tissues. Taken together, these results suggest that TZ-OSE can decrease anxiety, depression, and stress in vivo, regulate sleep-related factors, and control energy metabolism index factors. [ABSTRACT FROM AUTHOR]

The anti-inflammatory activity of benzopyran derivatives extracted from the Myagropsis myagroides against lipopolysaccharide (LPS)-induced RAW 264.7 murine macrophage cell line was examined. The three constituents of M. myagroides extracts were identified as the sargachromanol G, sargachromanol I, and saringosterol by 1H and 13C NMR. Sargachromanol G, sargachromanol I, and saringosterol at concentrations of 50 μg/mL showed inhibitory effects on LPS induced interleukin-6 production (by 84%, 90%, and 96%, respectively) and tumor necrosis factor (by 74%, 82%, and 98%, respectively). These results confirm the potential anti-inflammatory effects of sarga-chromanol G, sargachromanol I, and saringosterol. [ABSTRACT FROM AUTHOR]

Fueled by the recognition of hydrogen as a promising renewable energy source for the future, there have been many attempts to find greener and more economical ways for its production from various sources. In this study, Methylomonas sp. DH-1, a type I methanotroph, was found to produce hydrogen using methane as a sole carbon source, under micro-aerobic conditions; this is analogous to the partial oxidation of methane in a thermochemical process based on metal catalysts. Flask cultures of Methylomonas sp. DH-1 were used to investigate the effects of different culture conditions on hydrogen production, including oxygen levels, methane/oxygen ratios, and initial cell densities. Methylomonas sp. DH-1 could produce hydrogen at an oxygen level below 4%, regardless of the methane content in the flask, implying that the critical factor for hydrogen production is the oxygen level, rather than the methane/oxygen ratio. Moreover, Methylomonas sp. DH-1 shows reversibility in hydrogen production and uptake, because the strain produces hydrogen under micro-aerobic conditions, uptakes it when the oxygen levels increase, and restores the hydrogen production capability when conditions become microaerobic again. Under initial conditions of 30% methane, 70% air, and an OD600nm of 6, hydrogen production was 26.87 μmol and its yields per methane and dry cell weight were 14.98 mmol-H2/mol-CH4 and 101.53 μmol-H2/g DCW, respectively, after 24 h of cultivation. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus) produces a wide variety of toxins that cause various diseases in humans. S. aureus toxins are divided into three categories; (i) superantigens (SAgs) that interfere with receptor function and cause toxic shock syndrome (TSS) or staphylococcal food poisoning (SFP), (ii) exfoliative toxins (ETs) that destroy epidermal barrier functions and cause staphylococcal scalded skin syndrome (SSSS), and (iii) cytotoxins that cause cell lysis. Recently, anti-toxin neutralizing agents have been engineered and proven to be effective in preventing or treating staphylococcal diseases. However, neutralization of a single toxin may not sufficiently protect the host from S. aureus infection. Biofilm formation and the pathogenesis of S. aureus are closely associated with accessory gene regulator-quorum sensing (agr-QS) system that controls the expressions of many virulence factors including exotoxins. In addition to anti-toxin antibodies, indirect approaches involving targeting of the agr-QS system are considered a novel means of inhibiting the virulence of S. aureus and controlling various diseases caused by the pathogen. This review summarizes basic concepts of the pathogenic roles of S. aureus toxins, the molecular features of the agr-QS system, and of natural product-based anti-virulence therapies that target virulence, rather than growth. [ABSTRACT FROM AUTHOR]

Design and synthesis of a conjugate with high specificity and sensitivity constitutes a fundamental process for developing a highly sensitive immunoassay. In this study, we investigated the process of design, synthesis, and evaluation of gold nanoparticle (AuNP) conjugates that include both antibody and horseradish peroxidase (HRP) for application in a highly sensitive chemiluminescence immunoassay (hs-CLIA). To increase the labeling efficiency of HRP on the AuNP-based conjugates while maintaining process simplicity, two synthesis methods were suggested and evaluated using the physical adsorption-based conventional synthesis protocol. Specifically, the respective methods utilized adsorption or covalent coupling of aldehyde-activated (ald)HRP, which covalently binds to the primary amine group of a protein. The conjugates were characterized via spectroscopy and dynamic light scattering methods. Conjugate sensitivity was evaluated by not only analyzing the activity of HRP but also comparing the analytical sensitivity provided by the CL-based enzyme-linked immunosorbent assay for the detection of cardiac troponin I as a model target. Based on the results of this study, we demonstrated that the use of (ald)HRP for target labeling successfully enhanced the sensitivity of the AuNP-based conjugates; moreover, it could provide a promising labeling method for hs-CLIA. [ABSTRACT FROM AUTHOR]

Carotenoids, found in many fruits and vegetables, are antioxidants that protect human skin from UV radiation. In humans, fruit and vegetable intake increases carotenoid contents in skin, which are conventionally assessed by invasive blood tests. In this study, 47 healthy Korean subjects (volunteers) consumed fruit juice containing tomato, apple, strawberry, or grape three times per week for 6 weeks. Skin antioxidant levels were measured by non-invasive resonance Raman spectroscopy. The correlation between skin carotenoid (SC) score with demographic data (age, height, weight) and juice supplementation and changes in SC scores among groups were analyzed. Variations in skin antioxidant levels increased with juice supplementation (p < 0.05). Fruit juice intake was significantly correlated with SC score, indicating increased skin antioxidant levels. Grape and tomato increased skin antioxidant levels and showed higher antioxidant activity than other fruits. Fruit juices containing high levels of carotenoids and antioxidants may provide modest benefits to human health. [ABSTRACT FROM AUTHOR]

In recent years, nanofibers have been developed and widely used in many products, such as cosmetics and medical supplies. They can be fabricated from various synthetic or natural polymers and attached to bioactive compounds. In previous research, polycaprolactone (PCL) nanofibers containing Spirulina extract were demonstrated to be effective on dermal wound healing in a rat model. In this study, we fabricated Spirulina extract-alginate PCL nanofibers using alginate, which has hydrophilic structures capable of holding large amounts of water, to support the backbone of the nanofibers. The morphological characteristics, hydrophilicity, water absorbance, skin adhesiveness, toxicity to human keratinocyte cells (HaCaT), and Spirulina extract emission over time were assessed. Alginate improved the efficacy of Spirulina PCL nanofibers in moisture maintenance and adhesion ability, which highly affected recovery in the rat skin wound model. In conclusion, Spirulina extract-alginate PCL nanofibers could be considered a promising candidate for wound care. [ABSTRACT FROM AUTHOR]

Coal is one of the major sources of energy, fuel, and other related chemicals. The processes to utilize coal for energy, fuel and other chemicals such as coal combustion, liquefaction, carbonization, and gasification pose a great threat to the environment by emitting toxic particles and CO to the atmosphere. Thus, biological beneficiation of coal can be a good strategy to utilize coal with environmental sustainability. Here, we report the screening of microorganisms able to degrade or depolymerize coal. These host strains are potential candidates for the development of biological treatment process of coal. A total of 45 microbial strains were isolated from sludge enriched with coal and were identified based on 16S rRNA sequencing. Four strains of three genera, Cupriavidus sp., Pseudomonas sp., and Alcaligenes sp., were further characterized for their abilities to degrade coal. The degree of coal degradation was analyzed by measuring the increase in absorbance at 450 nm by UV spectroscopy. These microorganisms were also able to increase the pH of the culture media as a response to the acidic nature of coal. Laccase-like activity was also found in these strains when tested for RBBR dye degradation. Since biological degradation of coal through the use of microorganisms is a good alternative to chemical combustion of coal, microbial strains isolated in this study can be potential biological catalysts for coal conversion into valuable chemicals. [ABSTRACT FROM AUTHOR]

Organophosphate (OP) nerve agents are known as the most toxic chemical warfare agents that act by inhibiting the enzyme acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). Because BuChE is present at a level of about 3,900 times higher than AChE in plasma, most OP agents first react with BuChE in plasma, suggesting that OP-inactivated BuChE (OP-iBuChE) may act as a biomarker of OP exposure. In this study, we generated an anti-BuChE monoclonal antibody (mAb) that has reduced binding activity to VX-inactivated BuChE compared to native BuChE by phage display. We performed subtractive biopanning of a synthetic human Fab library against native BuChE and soman-iBuChE or VX-iBuChE. As the results, we isolated four Fab clones that showed differential binding activities to VX-iBuChE and native BuChE in ELISAs. To confirm the antigen-binding specificity of the selected clones, the Fabs were converted to IgG1s, and the IgG antibodies were expressed in HEK293F cells and purified. One of them (A2) showed approximately 30% reduced binding activity to VX-iBuChE compared to BuChE in a dose-dependent manner, whereas the other three antibodies showed almost the same binding activities to VX-iBuChE and BuChE. In addition, the A2 antibody did not show reduced binding activity to sarin-iBuChE or soman-iBuChE compared to native BuChE. The results indicate that A2 antibody shows reduced binding activity only to VX-iBuChE. A2 antibody may be applied to specific diagnosis of VX exposure. [ABSTRACT FROM AUTHOR]

The promoter region of copper-inducible laccase gene, LCC1, from Pycnoporus coccineus was explored in the heterologous expression of foreign protein in Pichia pastoris. The promoter region (P) was isolated and used to replace the methanol-inducible AOXI promoter (P) of pPICHOLI-2, an episomal expression vector for P. pastoris, to generate a new copper-inducible expression vector. The promoter activity of P was compared with those of P and P, a copper-inducible promoter of a commercial vector pPICHOLI-C, using a laccase gene as a reporter gene in P. pastoris GS115. Reporter laccase activity of the culture broth reached 182 and 43 units/L for P and P, respectively, after induction with 0.2 mM CuSO at OD = 1 and culture for 120 h at 15°C in complex medium containing 1% glucose. For P activity, yeast cells harboring P-laccase plasmid were cultured for 120 h at 15°C in complex medium with intermittent feeding with 1% methanol every 12 h to avoid methanol toxicity. Laccase activity of culture broth was 124 units/L. Conclusively, P is a new copper-inducible promoter that shows superior performance in terms of efficiency of laccase production compared to commercial vectors. P is additionally superior to P since it does not require laborious feeding with a carbon source. [ABSTRACT FROM AUTHOR]

Herbal medicinal products have proven to be safe, economical and effective alternatives to synthetic chemical pharmaceuticals. The green tea plant ( Camellia sinensis) is of profound medicinal value due to the presence of potent bioactive constituents. The purpose of the present work is to investigate saponins from green tea seeds for potential use as anti-angiogenic, antimicrobial, and hemolytic agents. Green tea seed saponins were separated into six fractions by reverse phase HPLC. The presence of three aglycone chains in the saponins of each fraction was confirmed by acid hydrolysis. Anti-angiogenic activity was evaluated using saponin fractions at concentrations in the range of 2.5 ~ 25 μg/mL. Antimicrobial activity was evaluated by thin-layer chromatography (TLC) using E. coli; S. mutans, a zoonotic Salmonella species and the fungal strain, A. niger. Saponin fractions were more potent against E. coli (gram negative bacteria) than against S. mutans (gram positive bacteria) and strongly inhibited six strains of zoonotic Salmonella. Green tea saponins also showed potent anti-angiogenic effects. All saponin fractions exhibited hemolytic activity. Our results confirm that green tea saponins have antimicrobial, anti-angiogenic, and hemolytic activities; indicating their potential as natural pharmaceutical products. [ABSTRACT FROM AUTHOR]

Camellia sinensis L. O. Kuntze has traditionally been used in Asian countries as a stimulant, a diuretic, and to prevent obesity. Studies in green tea are well-documented in literature, though the potential of fruit from the plant has never been studied in details. In the present study, the anti-adipogenic effect of aqueous extract of green tea seed cake extract (GTE) were analysed in-vitro and the anti-obesity effects were determined in-vivo in murine model fed high-fat and high-carbohydrate diets. We found that GTE inhibited the lipid accumulation in 3T3-L1 adipocyte and significantly lowered body weight, body fat-pad weights, blood lipid, glucose, insulin, and leptin levels in mice fed high-fat and high-carbohydrate diets. GTE suppressed the expression of important adipogenic genes (PPARγ, C/EBPα, SREBP, LPL, aP2 and leptin) and lipid metabolism genes (FAS). Furthermore, GTE induced the levels of adiponectin in serum and adipose tissue, as well as AMP-activated protein kinase (AMPK) in liver tissue. These results suggested that GTE ameliorates its anti-obesity effect through multiple cells signaling pathways in diets induced obesity in mice. [ABSTRACT FROM AUTHOR]

The commonly used trypan blue dye exclusion method and other modified cell viability methods, such as fluorescein dye and tetrazolium dye exclusion, artificially introduce toxic chemicals to cells and, thus, alter cellular organelles when measuring cell viability. Therefore, cell viability could be affected by the processes currently used to observe viability. In this study, the cell viability of Chinese hamster ovary (CHO) cells was measured by simply counting attached cells after the cultured CHO cells were attached on a Concanavalin A (Con A) substrate. The efficiency of cell attachment to Con A surfaces was different for live and dead cells allowing the cell viability of CHO cells to be measured without any chemical modifications to the cells. [ABSTRACT FROM AUTHOR]

Ethanol extracts of Psoraleae semen (PSE) are used in Korean folk medicine for tuberculostasis, nycturia, sarcoma, inflammation, and cancer. We investigated the utility of P. semen extract for the suppression of angiogenesis and adipogenesis. Eighteen natural products were screened using anti-angiogenesis experiments, and one candidate product was identified. Psoraleae semen provided dosedependent inhibition of angiogenesis, adipogenesis, and cell adhesion in vitro and weight loss in mice in vivo. The anti-angiogenenic efficacy of PSE was superior to that of epigallocatechin gallate (EGCG) obtained from green tea leaves. The expression of angiogenesis and adipogenesisrelated signal molecules was reduced by increasing the doses of PSE. A diet supplemented with PSE provided more effective weight loss in LB mice than did orlistat, a representative synthetic diet drug, which also caused hairloss in LB mice. P. semen may be a useful antiadipogenic compound if these in vitro results are replicated under in vivo conditions. [ABSTRACT FROM AUTHOR]

The fed-batch culture system was employed to enhance production of α-ketoglutarate (α-KG) by the strainsof Corynebacterium glutamicum, whose genes encoding the key enzymes responsible for the biosynthesis of L-glutamate from α-KG were deleted. In a shake flask fermentation, C. glutamicum JH110 in which the 3 genes, gdh (encoding glutamate dehydrogenase), gltB (encoding glutamate synthase), and aceA (encoding isocitrate lyase) were disrupted showed the highest production of α-KG (12.4 g/L) compared to the strains JH102 ( gdh mutant), JH103 ( gltB mutant), and JH107 ( gdh gltB double mutant). In the fed-batch cultures using a 5 L-jar fermenter, the strain JH107 produced more α-KG (19.5 g/L), but less glutamic acid (23.3 g/L) than those produced by the parent strain HH109, as well as JH102. The production of α-KG was significantly enhanced and the accumulation of glutamicacid was minimized by the ammonium-limited fed-batch cultures employing C. glutamicum JH107. Further improvement of α-KG production by the strain JH107 was achieved through the ammonium-limited fed-batch culture with the feeding of molasses, and the levels of α-KG and glutamic acid produced were 51.1 and 0.01 g/L, respectively. [ABSTRACT FROM AUTHOR]

In this study, we analyzed a bacterial community closely associated with Cochlodinium polykrikoides that caused harmful algal blooming in the sea. Filtration using a plankton mesh and percoll gradient centrifugation were performed to eliminate free-living bacteria. Attached bacteria were analyzed by culture-dependent and culture-independent methods. Five culturable bacterial strains were isolated and identified from the C. polykrikoides mixed bacterial community. The isolates belonged to α-Proteobacteria ( Nautella sp., Sagittula sp., and Thalassobius sp.) and γ-Proteobacteria ( Alteromonas sp. and Pseudoalteromonas sp.). All of the 5 isolates showed algicidal activity against C. polykrikoides and produced extracellular compounds responsible for algicidal properties after entering the stationary phase. The algicidal compounds produced by the 5 isolates were heat-stable and had molecular masses of less than 10,000 Da. Furthermore, the algicidal compounds were relatively specific for C. polykrikoides in terms of their algicidal activities. Culture-independent analysis of the bacterial community in association with C. polykrikoides was performed using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). On the basis of the PCR-DGGE profile, Sagittula sp. was identified as a dominant species in the bacterial community of C. polykrikoides. [ABSTRACT FROM AUTHOR]