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Development of facile polymerization routes and new monomers is attractive for the development of polymer chemistry and materials. This study integrates the dual concepts in one work. Design and synthesis of Meldrum's acid (MA)-amine compounds as monomers and their corresponding self-polymerization based on ketene-mediated chemistry have been reported. An effective synthetic route to polyamides under a melt process in 20 minutes is demonstrated. Self-polymerization of the MA-amine compounds results in both linear and hyperbranched polyamides with high molecular weights, good film formability, and high mechanical strength. Flexible polyamide films prepared through a solution casting method show a high tensile strength of 122 MPa and an elongation at break of 6.7%. The designed monomers and synthetic route open a new window to both polymer chemistry and high-performance polymer materials.

A major obstacle to nanodrugs-mediated cancer therapy is their rapid uptake by the reticuloendothelial system that decreases the systemic exposure of the nanodrugs to tumors and also increases toxicities. Intralipid has been shown to reduce nano-oxaliplatin-mediated toxicity while improving bioavailability. Here, we have found that Intralipid reduces the cytotoxicity of paclitaxel for human monocytic cells, but not for breast, lung, or pancreatic cancer cells. Intralipid also promotes the polarization of macrophages to the anti-cancer M1-like phenotype. Using a xenograft breast cancer mouse model, we have found that Intralipid pre-treatment significantly increases the amount of paclitaxel reaching the tumor and promotes tumor apoptosis. The combination of Intralipid with half the standard clinical dose of Abraxane reduces the tumor growth rate as effectively as the standard clinical dose. Our findings suggest that pre-treatment of Intralipid has the potential to be a powerful agent to enhance the tumor cytotoxic effects of Abraxane and to reduce its off-target toxicities.

Furan-functionalized Meldrum's acid (MAF) has been utilized as a monomer for the preparation of self-crosslinkable polymers which possess Meldrum's acid (MA) moieties as the reactive sites. As MAF contains acidic C–H and furan groups, both are reactive toward maleimide groups, and polymerization has been carried out between MAF and multi-functional maleimide compounds. Both linear and hyperbranched MA-containing self-crosslinkable polymers have been prepared. The prepared polymers show good solubility in aprotic high polar solvents and high film formability. Self-standing and highly flexible crosslinked films have been obtained. The polymer film shows high solvent resistance, a high glass transition temperature of 298 °C, a high thermal stability above 340 °C, and attractive mechanical properties with a Young's modulus of 3.58 GPa and a stress at break of 34.4 MPa. Moreover, the crosslinked polymer films have relatively low-dielectric constants of about 2.85–2.78. An effective approach has been demonstrated for the preparation of high molecular weight MA-containing polymers and the corresponding thermosetting resin based self-standing and flexible polymer films.

Experimental studies were done with a small pipe with a diameter of 0.043 m and a large pipe with a diameter of 0.49 m to demonstrate the flame propagation suppression with inertia isolation in a long duct. Tests were carried in an ignition section containing propylene/air mixture near stoichiometric concentration and generating a peak flame propagation speed of approximately 100 m/s. The ignition section is connected to a section filled with an inert gas, another section with flammable mixtures, and finally a sufficiently long, ambient section to accommodate flame propagation. The critical length of the inert gas section required for successful suppression of flame from the igniting the flammable section is found to be 0.6 m for CO2 and 0.9 m for N2 in the large pipe and 0.2 m for CO2 and 0.3 m for N2 in the small pipe. Additional tests with a 3 m of ignition section and peak flame propagation speed of 225 m/s showed that the critical length for successful suppression by CO2 is only increased slightly to 0.9 m, confirming that the suppression is a result of inertia isolation rather than inert gas dilution. Finally, application of the results in responding to large-scale leak into a long, underground duct is discussed.

Meldrum’s acid (MA) is utilized as a monomer to polymerize with diisocyanates to result in polyamides, containing MA moieties at polymer chains. This reaction is also employed to prepare isocyanate-terminated polyamide segments which are utilized as a precursor for preparation of MA-containing polyurethanes. Based on the thermolysis reaction of MA groups, followed by ketene dimerization reaction, the reactive polyamides and polyurethanes show self-cross-linkable features. The cross-linked polyurethanes exhibit good film formability, thermal stability, and mechanical properties. A new MA-based polymerization method and a novel synthesis route for preparation of reactive polyamides and polyurethanes are demonstrated.

In this work, the shock sensitivity of hexachlorodisilane (HCDS) hydrolysis products was studied. The hydrolysis conditions included vapor and liquid HCDS hydrolysis in moist air. Shock sensitivity was determined by using a Fall hammer apparatus. Extensive infrared studies were done for the hydrolysis products. It was found that the Si–Si bond in HCDS during hydrolysis is preserved and can be cleaved by shock, leading to intramolecular oxidation of the neighboring silanol (Si–OH) groups to form a networked Si–O–Si structure and hydrogen gas. The limiting impact energy for shock sensitivity was also found proportional to the oxygen/silicon ratio in the deposit. Finally, recommendations are given for controlling the shock sensitivity of the hydrolyzed deposit.

Meldrum's acid (MA) could be considered as a precursor of a highly reactive ketene group, and consequently is a suitable group for building up crosslinkable small molecular compounds and polymer chains. In this work, the Michael addition reaction of MA is applied for the preparation of multifunctional MA-containing compounds, exploring an effective route to synthesize multifunctional MA compounds, used as monomers for the corresponding MA-based thermosetting resins. Moreover, MA is utilized as a difunctional monomer to be polymerized with a bismaleimide (BMI) through the Michael addition reaction. A polymerization method directly using MA as a monomer for the synthesis of reactive and crosslinkable MA-containing polymers (poly(BMI-MA)) is demonstrated. The corresponding crosslinked CR-poly(BMI-MA) sample shows a high glass transition temperature (315 °C), high mechanical strength and a low dielectric constant of 3.15 at 1 MHz. A new type of MA reactive route and the corresponding high performance crosslinked resins are demonstrated.

Bone tissue engineering holds great promise and clinical efficacy for the regeneration of bone defects. In this study, an amphoteric N,O-carboxymethyl chitosan (NOCC) and fucoidan (FD) were covalently cross-linked via an amidation reaction to synthesize NOCC/FD composite hydrogels. The hydrogels were lyophilized and then three-dimensional scaffolds with interconnected macropores were obtained. To enhance the mechanical properties and osteogenic activity, the NOCC/FD scaffolds were biomineralized for the growth of hydroxyapatite crystals. A comparative assessment of the structures, morphologies, and physical properties of the original and mineralized scaffolds were performed by SEM, EDS, X-ray diffraction and FT-IR analysis. FD regulated the growth of hydroxyapatite nanocrystallites (n-HAp) and thus the NOCC/FD scaffolds showed better mineralization efficiency than NOCC scaffolds. The compressive strength of the scaffolds was greatly enhanced after mineralization with n-HAp. The n-HAp/NOCC/FD scaffolds enhanced the proliferation, ALP activity, and mineralization of osteoblast cells more strongly than the original and mineralized NOCC scaffolds. Hence, the n-HAp-mineralized NOCC/FD scaffolds may prove to be an excellent and versatile scaffold for bone tissue engineering.

Nitroxide-functionalized polymers are attractive macroinitiators for nitroxide mediated polymerization (NMP) and further modification of polymers. In this work, a facile reaction method to chemically bond nitroxide groups to polymer chains has been demonstrated with incorporation of 2,2,6,6-tetramethylpiperydinyl-1-oxyl (TEMPO) groups into poly(2,6-dimethyl-1,4-phenylene oxide) (PPO) through the atom transfer radical coupling reaction. The TEMPO-functionalized PPO (PPO-TEMPO) has been used as a macroinitiator for NMP of styrene to result in the corresponding PPO-g-PS graft copolymers. Moreover, PPO-TEMPO could be used to carry out a thermally-induced self-crosslinking reaction through radical coupling. The crosslinked PPO displays enhanced mechanical properties, reduced oxygen permeability coefficient (from 22.5 barrer to 3.5 barrer), low dielectric constant (2.40 at 10 GHz) and low dissipation factor (0.008 at 10 GHz), so as to have high potential for application in dielectric layers of microelectronics.

A universal tetherable initiator, derived from the structure of fatty acids, for surface-initiated atom transfer radical polymerization (SI-ATRP) from metal oxide surfaces was prepared. A simple amidation between 2-bromoisobutyryl bromide and ω-aminolauric acid allowed preparation of 12-(2-bromoisobutyramido)dodecanoic acid (BiBADA). After facile purification, BiBADA was used as a tetherable initiator for a broad range of metal oxide nanoparticles. The modified nanoparticles were grafted with methyl methacrylate or n-butyl acrylate via SI-ATRP with a high grafting density. This is the first report of successful SI-ATRP from a selection of different metal oxide nanoparticles. Sub-10 nm Fe3O4 nanoparticles with an intrinsically tethered initiator were also prepared using BiBADA as a surfactant template. Additional experiments demonstrated successful modification of an aluminum foil surface with polymer brushes using BiBADA as a tetherable initiator.

Clinically, high levels of acrolein (a highly reactive α, β-unsaturated aldehyde) and acrolein adducts are detected in the brain of patients with CNS neurodegenerative diseases, including Alzheimer’s disease and spinal cord injury. Our previous study supports this notion by showing acrolein as a neurotoxin in a Parkinsonian animal model. In the present study, the effect of AZD6244 (an ATP non-competitive MEK1/2 inhibitor) on acrolein-induced neuroinflammation was investigated using BV-2 cells and primary cultured microglia. Our immunostaining study showed that lipopolysaccharide (LPS, an inflammation inducer as a positive control) increased co-localized immunoreactivities of phosphorylated ERK and ED-1 (a biomarker of activated microglia) in the treated BV-2 cells. Similar elevation in co-localized immunoreactivities of phosphorylated ERK and ED-1 was detected in the acrolein-treated BV-2 cells. Furthermore, Western blot assay showed increases in phosphorylated ERK in BV-2 cells subjected to LPS (1 μg/mL) or acrolein (30 μM); these increases were blocked by AZD6244 (10 μM). At the same time, AZD6244 attenuated LPS-induced TNF-α (a pro-inflammatory cytokine) and cyclooxygenase-II (COX II, a pro-inflammatory enzyme). Consistently, AZD6244 reduced acrolein-induced elevations in COX-II mRNA and COX-II protein expression. In addition, AZD6244 inhibited acrolein-induced increases in activated caspase 1 (a biomarker of inflammasome activation) and heme oxygenase-1 (a redox-regulated chaperone protein) in BV-2 cells. Using a transwell migration assay, AZD6244 attenuated acrolein (5 μM)-induced migration of BV-2 cells and primary cultured microglia. In conclusion, our study shows that acrolein is capable of inducing neuroinflammation which involved ERK activation in microglia. Furthermore, AZD6244 is capable of inhibiting acrolein-induced neuroinflammation. Our study suggests that ERK inhibition may be a neuroprotective target against acrolein-induced neuroinflammation in the CNS neurodegenerative diseases.

Background and purpose Ischemic stroke is the most common type of cerebrovascular event and is responsible for approximately 85% of all strokes in Taiwan. Neurons contain high concentrations of polyamines, which are prone to various pathological states in the brain and are perturbed after cerebral ischemia. Acrolein, an α,β-unsaturated aldehyde, has been suggested as the primary culprit of neuronal damage in stroke patients. However, the mechanism by which acrolein induces neuronal damage during ischemic stroke is not clear. Methods Urinary 3-hydroxypropyl mercapturic acid (3-HPMA), an acrolein-glutathione (GSH) metabolite, plasma acrolein-protein conjugates (Acr-PC) and plasma GSH levels were analyzed to correlate disease severity and prognosis of stroke patients compared with control subjects. In vivo middle cerebral artery occlusion (MCAO) animal models and an in vitro oxygen glucose deprivation (OGD) stroke model were used to investigate the mechanisms of acrolein-induced neuronal damage. Results A deregulated acrolein metabolism, including significantly increased plasma Acr-PC levels, decreased urinary 3-HPMA levels and decreased plasma GSH levels, was found in stroke patients compared to control subjects. We further observed that acrolein was produced during ischemia resulting in brain damage in in vivo MCAO animal model. The induction of acrolein in neuronal cells during OGD occurred due to the increased expression of spermidine/spermine N1-acetyltransferase (SSAT) by NF-kB pathway activation. In addition, acrolein elicited a vicious cycling of oxidative stress resulting in neurotoxicity. Finally, N-acetylcysteine effectively prevented OGD-induced neurotoxicity by scavenging acrolein. Conclusion Overall, our current results demonstrate that acrolein is a culprit of neuronal damage through GSH depletion in stroke patients. The mechanism underlying the role of acrolein in stroke-related neuronal damage occurs through SSAT-induced polyamine oxidation by NF-kB pathway activation. These results provide a novel mechanism of neurotoxicity in stroke patients, aid in the development of neutralizing or preventive measures, and further our understanding of neural protection.

Platelet-rich plasma (PRP) is rich in growth factors and has commonly been utilized in&nbsp, the repair and regeneration of damaged articular cartilage. However, the major drawbacks of direct PRP injection are unstable biological fixation and fast or burst release of growth factors. Fucoidan is a heparinoid compound that can bind growth factors to control their release rate. Furthermore, fucoidan can reduce arthritis through suppressing inflammatory responses and thus it has been reported to prevent the progression of osteoarthritis, promote bone regeneration and accelerate healing of cartilage injury. Injectable hydrogels can be used to deliver cells and growth factors for an alternative, less invasive treatment of cartilage defects. In this study, hyaluronic acid (HA) and fucoidan (FD) was blended with gelatin (GLT) and the GLT/HA/FD hybrid was further cross-linked with genipin (GP) to prepare injectable GP-GLT/HA/FD hydrogels. The gelation rate was affected by the GP, GLT, HA and FD concentrations, as well as the pH values. The addition of HA and FD to GLT networks improved the mechanical strength of the hydrogels and facilitated the sustained release of PRP growth factors. The GP-GLT/HA/FD hydrogel showed adequate injectability, shape-persistent property and strong adhesive ability, and was more resistant to enzymatic degradation. The PRP-loaded GP-GLT/HA/FD hydrogel promoted cartilage regeneration in rabbits, which may lead to an advanced PRP therapy for enhancing cartilage repair.

Casticin, a polymethoxyflavone occurring in natural plants, has been shown to have anticancer activities. In the present study, we aims to investigate the anti-skin cancer activity of casticin on melanoma cells in vitro and the antitumor effect of casticin on human melanoma xenografts in nu/nu mice in vivo. A flow cytometric assay was performed to detect expression of viable cells, cell cycles, reactive oxygen species production, levels of [Formula: see text] and caspase activity. A Western blotting assay and confocal laser microscope examination were performed to detect expression of protein levels. In the in vitro studies, we found that casticin induced morphological cell changes and DNA condensation and damage, decreased the total viable cells, and induced G2/M phase arrest. Casticin promoted reactive oxygen species (ROS) production, decreased the level of [Formula: see text], and promoted caspase-3 activities in A375.S2 cells. The induced G2/M phase arrest indicated by the Western blotting assay showed that casticin promoted the expression of p53, p21 and CHK-1 proteins and inhibited the protein levels of Cdc25c, CDK-1, Cyclin A and B. The casticin-induced apoptosis indicated that casticin promoted pro-apoptotic proteins but inhibited anti-apoptotic proteins. These findings also were confirmed by the fact that casticin promoted the release of AIF and Endo G from mitochondria to cytosol. An electrophoretic mobility shift assay (EMSA) assay showed that casticin inhibited the NF-[Formula: see text]B binding DNA and that these effects were time-dependent. In the in vivo studies, results from immuno-deficient nu/nu mice bearing the A375.S2 tumor xenograft indicated that casticin significantly suppressed tumor growth based on tumor size and weight decreases. Early G2/M arrest and mitochondria-dependent signaling contributed to the apoptotic A375.S2 cell demise induced by casticin. In in vivo experiments, A375.S2 also efficaciously suppressed tumor volume in a xenotransplantation model. Therefore, casticin might be a potential therapeutic agent for the treatment of skin cancer in the future.

Hexachlorodisilane (Si2Cl6, HCDS) is an important precursor used in semiconductor device manufacturing. It is a flammable as well as a water reactive liquid which hydrolyzes rapidly upon contact with water or moisture. The hydrolyzed deposits are also known to be shock-sensitive with explosion energy equivalent to trinitrotoluene (TNT). In this work, two phases of test program including disposal of HCDS and disposal of the shock sensitive HCDS hydrolyzed deposits were conducted. The first phase of the program was to find an agent that can completely dissolve/react the HCDS vapor without forming shock sensitive deposits. The second phase of the program attempted to find a suitable agent to suppress the Si–Si bonds, one of the essential roles of chemical functional groups in shock sensitivity of the HCDS hydrolyzed deposits to suppress the shock sensitivity. A variety of agents such as sulfuric acid solutions, aqueous sodium hydroxide (NaOH) solutions, aqueous potassium hydroxide (KOH) solutions, KOH/alcohol solutions were utilized as the suppressants in this work. Samples mixed with suppressants were not only tested for shock sensitivity by a Fall-hammer apparatus but also analyzed for chemical functional groups to identify the effect of each agent. Concentrated sulfuric acid was found to suppress the shock sensitivity of the liquid HCDS hydrolyzed deposits by acting as a medium that helps the hydrolyzed deposit to retain moisture. KOH/alcohol solutions can turn HCDS vapor into non-hazardous silica, so that, it provided a safe way to dispose HCDS. Finally, practical recommendations about handling and eliminating the risk of shock sensitivity are given for HCDS liquid spill, HCDS vapor vent and HCDS hydrolyzed deposits.

Hydroxypropyl chitosan (HPCS) has recently attracted increasing attention in biomedical applications because it has enhanced water solubility, excellent biocompatibility, and better antioxidant and antibacterial activities compared with chitosan. However, HPCS doesn't meet the mechanical strength requirement in bone tissue engineering and is not suitable for cell adhesion and growth because of its hydrophilic nature and low crystallinity. In this study, nano-scaled hydroxyapatite (n-HA) and HPCS were synthesized, respectively, and then n-HA/HPCS nanocomposite scaffolds were developed by incorporating n-HA into HPCS matrix accompanied with crosslinking of HPCS by a naturally occurring compound, genipin (GP), which in turn greatly altered the hydrophilicity and mechanical properties. The nanocomposite scaffolds showed an open structure with interconnected pores and a rough morphology with n-HA inserted in the GP-crosslinked HPCS matrix. The porosity, swelling capacity, compressive strength, fluorescence emission and degradation rate can be regulated by varying GP concentrations and n-HA contents. An osteoconductive and osteogenic marine algae polysaccharide, fucoidan, was further adsorbed to the composite scaffolds via electrostatic interactions. Incorporation of n-HA and adsorption of FD into the composite scaffolds increased ALP activity in 7F2 osteoblast cells and promoted their mineralization. The FD-adsorbed n-HA/HPCS composite scaffolds can be a potential biomaterial for BTE applications.

Oral administration is a highly attractive approach for the delivery of protein drugs. However, oral protein therapeutics typically exhibit extremely poor bioavailability due to the harsh gastrointestinal (GI) environments and low permeability of protein across the intestinal barrier. Trimethyl chitosan (TMC) shows excellent mucoadhesive and absorption-enhancing properties while fucoidan (FD) has hypoglycemic effects and can prevent diabetes-related complications. Here we report, for the first time, that TMC combined with FD can be developed to a mutlifunctional nanoplatform for enhancing the transepithelial permeation of insulin through the intestinal epithelial cell barrier and inhibiting the α-glucosidase activity. TMC and FD self-assembled into spherical nanoparticles (NPs) for insulin encapsulation. TMC/FD NPs protected insulin against degradation by releasing insulin in a pH-dependent manner in the gastrointestinal tract fluids. The NPs were able to modulate the barrier function of the Caco-2 intestinal epithelial cell monolayer, and enhance paracellular transport of insulin across the intestinal barrier. TMC/FD NPs also showed α-glucosidase inhibitory activity, with an inhibition ratio of 33.2% at 2 mg/mL. The superior transepithelial absorption enhancing property of the TMC/FD NPs is expected to combine in the future with the functions of fucoidan against diabetes-related complications for development of advanced mutlifunctional therapeutic platforms for diabetes.

Self-assembled nanoparticles (NPs) from arginine-modified chitosan (CS-N-Arg) and thiolated fucoidan (THL-fucoidan) were synthesized to enhance the transport of dextran and curcumin across intestinal epithelial cell layer. CS-N-Arg/THL-fucoidan NPs exhibited a pH-sensitive assembly-disassembly and drug release property. Evaluations of the NPs in enhancing the transport of a hydrophilic macromolecule (FITC-dextran) and a hydrophobic drug (curcumin) were investigated in Caco-2 cell monolayers. The cationic CS-N-Arg in the NPs induced disruption of intestinal epithelial tight junctions as indicated by the decrease of transepithelial electrical resistance (TEER). Permeation studies revealed that the NPs enhanced the paracellular permeation of macromolecular dextran through the monolayer barrier. In addition, the multifunctional NPs increased the permeability of rhodamine 123 because the thiomer THL-fucoidan in the NPs inhibited P-glycoprotein. Cellular uptake and permeability of curcumin encapsulated in the NPs were improved due to increasing their water solubility and stability.

The unliganded tetrameric Hb S has axial and lateral contacts with neighbors and can polymerize in solution. Novel recombinants of Hb S with single amino acid substitutions at the putative axial (recombinant Hb (rHb) (βE6V/αH20R) and rHb (βE6V/αH20Q)) or lateral (rHb (βE6V/αH50Q)) or double amino acid substitutions at both the putative axial and lateral (rHb (βE6V/αH20R/αH50Q) and rHb (βE6V/αH20Q/αH50Q)) contact sites were expressed in Escherichia coli and purified for structural and functional studies. The (1)H NMR spectra of the CO and deoxy forms of these mutants indicate that substitutions at either αHis-20 or αHis-50 do not change the subunit interfaces or the heme pockets of the proteins. The double mutants show only slight structural alteration in the β-heme pockets. All mutants have similar cooperativity (n50), alkaline Bohr effect, and autoxidation rate as Hb S. The oxygen binding affinity (P50) of the single mutants is comparable with that of Hb S. The double mutants bind oxygen with slightly higher affinity than Hb S under the acidic conditions. In high salt, rHb (βE6V/αH20R) is the only mutant that has a shorter delay time of polymerization and forms polymers more readily than Hb S with a dextran-Csat value of 1.86 ± 0.20 g/dl. Hb S, rHb (βE6V/αH20Q), rHb (βE6V/αH50Q), rHb (βE6V/αH20R/αH50Q), and rHb (βE6V/αH20Q/αH50Q) have dextran-Csat values of 2.95 ± 0.10, 3.04 ± 0.17, 11.78 ± 0.59, 7.11 ± 0.66, and 10.89 ± 0.83 g/dl, respectively. rHb (βE6V/αH20Q/αH50Q) is even more stable than Hb S under elevated temperature (60 °C).

// Yu-Tang Tung 1, * , Pin-Wu Huang 1, * , Yu-Ching Chou 1 , Cheng-Wei Lai 1 , Hsiu-Po Wang 1 , Heng-Chien Ho 2 , Chih-Ching Yen 1, 3 , Chih-Yen Tu 1, 3 , Tung-Chou Tsai 1 , Dah-Cherng Yeh 4 , Jiun-Long Wang 1, 5 , Kowit-Yu Chong 6, 7 , Chuan-Mu Chen 1, 8 1 Department of Life Sciences, and Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan 2 Department of Medicine, China Medical University Hospital, Taichung 404, Taiwan 3 Department of Internal Medicine, China Medical University Hospital, Taichung 404, Taiwan 4 Department of General Surgery and Department of Internal Medicine, Taichung Veterans General Hospital, Taichung 407, Taiwan 5 Division of Chest Medicine, Department of Internal Medicine, Taichung Veterans General Hospital, Taichung 407, Taiwan 6 Department of Medical Biotechnology and Laboratory Sciences, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan 7 Molecular Medicine Research Center, College of Medicine, Chang Gung University, Tao-Yuan 333, Taiwan 8 Rong-Hsing Translational Medicine Center and iEGG Center, National Chung Hsing University, Taichung 402, Taiwan * These authors have contributed equally to this work Correspondence to: Chuan-Mu Chen, e-mail: chchen1@dragon.nchu.edu.tw Keywords: VEGF, transgenic mice, pulmonary tumorigenesis, magnetic resonance imaging (MRI), miRNA therapy Received: November 03, 2014 Accepted: April 10, 2015 Published: April 22, 2015 ABSTRACT Many studies have shown that vascular endothelial growth factor (VEGF), especially the human VEGF-A 165 (hVEGF-A 165 ) isoform, is a key proangiogenic factor that is overexpressed in lung cancer. We generated transgenic mice that overexpresses hVEGF-A 165 in lung-specific Clara cells to investigate the development of pulmonary adenocarcinoma. In this study, three transgenic mouse strains were produced by pronuclear microinjection, and Southern blot analysis indicated similar patterns of the foreign gene within the genomes of the transgenic founder mice and their offspring. Accordingly, hVegf-A 165 mRNA was expressed specifically in the lung tissue of the transgenic mice. Histopathological examination of the lung tissues of the transgenic mice showed that hVEGF-A 165 overexpression induced bronchial inflammation, fibrosis, cysts, and adenoma. Pathological section and magnetic resonance imaging (MRI) analyses demonstrated a positive correlation between the development of pulmonary cancer and hVEGF expression levels, which were determined by immunohistochemistry, qRT-PCR, and western blot analyses. Gene expression profiling by cDNA microarray revealed a set of up-regulated genes ( hvegf-A 165 , cyclin b1 , cdc2 , egfr , mmp9 , nrp-1 , and kdr ) in VEGF tumors compared with wild-type lung tissues. In addition, overexpressing hVEGF-A 165 in Clara cells increases CD105, fibrogenic genes ( collagen α1 , α-SMA , TGF-β1 , and TIMP1 ), and inflammatory cytokines (IL-1, IL-6, and TNF- α ) in the lungs of hVEGF-A 165 -overexpressing transgenic mice as compared to wild-type mice. We further demonstrated that the intranasal administration of microRNA-16 (miR-16) inhibited lung tumor growth by suppressing VEGF expression via the intrinsic and extrinsic apoptotic pathways. In conclusion, hVEGF-A 165 transgenic mice exhibited complex alterations in gene expression and tumorigenesis and may be a relevant model for studying VEGF-targeted therapies in lung adenocarcinoma.

Piperlongumine (PL), a natural product of Piper longum, inhibits multiple malignant phenotypes. Therefore, the present study examined whether PL suppresses cancer stemness in oral cancer. The cellular effects of PL were determined by examining alterations in tumor sphere formation, cell migration, invasion, proliferation ability, chemosensitivity and radiosensitivity. Reverse transcription-quantitative polymerase chain reaction analysis and western blotting were performed in order to determine molecular expression levels. The present study revealed that PL inhibited cancer stem cell-forming ability and suppressed the expression of the stemness-related transcription factors SRY-Box 2, POU class 5 homeobox 1, and Nanog homeobox. However, it increased the expression of the differentiation marker cytokeratin 18. PL also suppressed cell migration and invasion, resulting in the elimination of the epithelial-mesenchymal transition. Furthermore, PL increased chemo- and radiosensitivity and suppressed tumor growth in vitro and in vivo. The results of the present study suggested that PL inhibits malignant phenotypes via the suppression of cancer stemness in oral cancer. Thus, PL may serve as an effective therapeutic agent for oral cancer.

Numerous reports illustrate the diverse effects of chewing the areca nut, most of which are harmful and have been shown to be associated with oral cancer. Nearly all of the studies are focused on the extract and/or low molecular weight ingredients in the areca nut. The purpose of this report is to identify the major protein component in the areca nut. After ammonium sulfate fractionation, the concentrated areca nut extract is subjected to DEAE-cellulose chromatography. A colored protein is eluted at low NaCl concentration and the apparently homogeneous eluent represents the major protein component compared to the areca nut extract. The colored protein shares partial sequence identity with the royal palm tree peroxidase and its peroxidase activity is confirmed using an established assay. In the study, the natural substrates of areca nut peroxidase are identified as catechin, epicatechin, and procyanidin B1. The two former substrates are similarly oxidized to form a 576 Da product with concomitant removal of four hydrogen atoms. Interestingly, oxidation of procyanidin B1 occurs only in the presence of catechin or epicatechin and an additional product with an 864 Da molecular mass. In addition, procyanidin B1 is identified as a peroxidase substrate for the first time.

The haemoglobin (Hb) of the extinct woolly mammoth has been recreated using recombinant genes expressed inEscherichia coli. The globin gene sequences were previously determined using DNA recovered from frozen cadavers. Although highly similar to the Hb of existing elephants, the woolly mammoth protein shows rather different responses to chloride ions and temperature. In particular, the heat of oxygenation is found to be much lower in mammoth Hb, which appears to be an adaptation to the harsh high-latitude climates of the Pleistocene Ice Ages and has been linked to heightened sensitivity of the mammoth protein to protons, chloride ions and organic phosphates relative to that of Asian elephants. To elucidate the structural basis for the altered homotropic and heterotropic effects, the crystal structures of mammoth Hb have been determined in the deoxy, carbonmonoxy and aquo-met forms. These models, which are the first structures of Hb from an extinct species, show many features reminiscent of human Hb, but underline how the delicate control of oxygen affinity relies on much more than simple overall quaternary-structure changes.

The carbohydrate polymer, hyaluronan, is a major component of the extracellular matrix in animal tissues. Exogenous hyaluronan has been used to treat osteoarthritis (OA), a degenerative joint disease involving inflammatory changes. The underlying mechanisms of hyaluronan in OA are not fully understood. Pro-inflammatory interleukin (IL)-1β downregulates peroxisome proliferator-activated receptor gamma (PPARγ), and increases expression of matrix metalloproteinases (MMPs) which are responsible for the degeneration of articular cartilage. The effects of low- and high-molecular-weight hyaluronan (oligo-HA and HMW-HA) on the inflammatory genes were determined in human SW-1353 chondrosarcoma cells. HMW-HA antagonized the effects of IL-1β by increasing PPARγ and decreasing cyclooxygenase (COX)-2, MMP-1, and MMP-13 levels. It promoted Akt, but suppressed mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NFκB) signaling, indicating anti-inflammatory effects. In contrast, the cells had overall opposite responses to oligo-HA. In conclusion, HMW-HA and oligo-HA exerted differential inflammatory responses via PPARγ in IL-1β-treated chondrosarcoma cells.

The purpose of this study was to identify the genes induced early in murine oral carcinogenesis. Murine tongue tumors induced by the carcinogen, 4-nitroquinoline 1-oxide (4-NQO), and paired non-tumor tissues were subjected to microarray analysis. Hierarchical clustering of upregulated genes in the tumor tissues revealed an association of induced genes with inflammation. Cytokines/cytokine receptors induced early were subsequently identified, clearly indicating their involvement in oral carcinogenesis. Hierarchical clustering also showed that cytokine-mediated inflammation was possibly linked with Mapk6. Cox2 exhibited the greatest extent (9-18 fold) of induction in the microarray data, and its early induction was observed in a 2h painting experiment by RT-PCR. MetaCore analysis showed that overexpressed Cox2 may interact with p53 and transcriptionally inhibit expression of several downstream genes. A painting experiment in transgenic mice also demonstrated that NF-κB activates early independently of Cox2 induction. MetaCore analysis revealed the most striking metabolic alterations in tumor tissues, especially in lipid metabolism resulting from the reduction of Pparα and Rxrg. Reduced expression of Mapk12 was noted, and MetaCore analysis established its relationship with decreased efficiency of Pparα phosphorylation. In conclusion, in addition to cytokines/cytokine receptors, the early induction of Cox2 and NF-κB activation is involved in murine oral carcinogenesis.

To elevate chemo-resistance of human cancer cells is a major obstacle in the treatment and management of malignant cancers. Diallyl sulfide (DAS), diallyl disulfide (DADS) and diallyl trisulfide (DATS) are presented in the Alliaceae family particularly in garlic. Although DAS, DADS and DATS have been shown to exhibit anticancer activities, there is little information on effects of these compounds on drug resistant genes in human colon cancer cells in vitro and in vivo. Herein, we are the first to show that DAS, DADS and DATS at 25 μM for 24-h and 48-h incubations promoted expression of drug resistant genes in colo 205 human colon cancer cells. In vitro experiments indicated that DATS promoted gene expression of multidrug resistant 1 (Mdr1) (p

Our earlier studies have demonstrated that gallic acid (GA) induced cytotoxic effects including induction of apoptosis and DNA damage and inhibited the cell migration and invasion in human cancer cells. However, GA-affected DNA damage and repair gene expressions in human prostate cancer cells are still unclear. In this study, we investigated whether or not GA induces DNA damage and inhibits DNA repair gene expression in a human prostate cancer cell line (PC-3). The results from flow cytometric assay indicated that GA decreased the percentage of viable PC-3 cells in a dose- and time-dependent manner. PC-3 cells after exposure to different doses (50, 100, and 200 μM) of GA and various periods of time (12, 24, and 48 h) led to a longer DNA migration smear (comet tail) occurred based on the single cell gel electrophoresis (comet assay). These observations indicated that GA-induced DNA damage in PC-3 cells, which also confirmed by 4,6-diamidino-2-phenylindole dihydrochloride staining and DNA agarose gel electrophoresis. Alternatively, results from real-time polymerase chain reaction assay also indicated that GA inhibited ataxia telangiectasia mutated, ataxia-telangiectasia and Rad3-related, O⁶-methylguanine-DNA methyltransferase, DNA-dependent serine/threonine protein kinase, and p53 mRNA expressions in PC-3 cells. Taken together, the present study showed that GA caused DNA damage and inhibited DNA repair genes as well as both effects may be the critical factors for GA-inhibited growth of PC-3 cells in vitro.

Curcumin from the rhizome of the Curcuma longa plant has been noted for its chemo-preventative and chemo-therapy activities, and it inhibits the growth of many types of human cancer cell lines. In this study, the mechanisms of cell death involved in curcumin-induced growth inhibition, including cell cycle arrest and induction of apoptosis in human tongue cancer SCC-4 cells, were investigated. Herein, we observed that curcumin inhibited cell growth of SCC-4 cells and induced cell death in a dose-dependent manner. Treatment of SCC-4 cells with curcumin caused a moderate and promoted the G(2) /M phase arrest, which was accompanied with decreases in cyclin B/CDK1 and CDC25C protein levels. Moreover, curcumin significantly induced apoptosis of SCC-4 cells with a decrease of the Bcl-2 level, reduction of mitochondrial membrane potential (ΔΨ(m) ), and promoted the active forms of caspase-3. Curcumin also promoted the releases of AIF and Endo G from the mitochondria in SCC-4 cells by using confocal laser microscope. Therefore, we suggest that curcumin induced apoptosis through a mitochondria-dependent pathway in SCC-4 cells. In addition, we also found that curcumin-induced apoptosis of SCC-4 cells was partly through endoplasmic reticulum stress. In conclusion, curcumin increased G(2) /M phase arrest and induced apoptosis through ER stress and mitochondria-dependent pathways in SCC-4 cells.

Many anticancer drugs are obtained from phytochemicals and natural products. However, some phytochemicals have mutagenic effects. Safrole, a component of Piper betle inflorescence, has been reported to be a carcinogen. We have previously reported that safrole induced apoptosis in human oral cancer cells in vitro and inhibited the human oral tumor xenograft growth in vivo. Until now, there is no information addressing if safrole promotes immune responses in vivo. To evaluate whether safrole modulated immune function, BALB/c mice were intraperitoneally injected with murine myelomonocytic WEHI-3 leukemia cells to establish leukemia and then were treated with or without safrole at 4 and 16 mg/kg. Animals were sacrificed after 2 weeks post-treatment with safrole for examining the immune cell populations, phagocytosis of macrophages and the natural killer (NK) cells' cytotoxicity. Results indicated that safrole increased the body weight, and decreased the weights of spleen and liver in leukemic mice. Furthermore, safrole promoted the activities of macrophages phagocytosis and NK cells' cytotoxicity in leukemic mice when compared with untreated leukemic mice. After determining the cell marker population, we found that safrole promoted the levels of CD3 (T cells), CD19 (B cells) and Mac-3 (macrophages), but it did not affect CD11b (monocytes) in leukemic mice. In conclusion, safrole altered the immune modulation and inhibited the leukemia WEHI-3 cells in vivo.

Diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS) are major organosulfur compounds exiting in garlic (Allium sativum). These compounds are reported to exhibit various pharmacological properties such as antibacteria, antiangiogenesis, anticancer, and anticoagulation, and they also induce cytotoxicity and induction of apoptosis in human cancer cells. Although these compounds show wide spectrum of biological activities, there are no reports to show that DAS, DADS, and DATS affected migration and invasion of human colon cancer cells, and their exact molecular mechanisms are not well investigated. Therefore, the purpose of this study was to determine whether DAS, DADS, and DATS affected the invasion and migration abilities of colo 205 human colon cancer cells. The results indicate that DAS, DADS, and DATS at 10 and 25 μM inhibited the migration and invasion of colo 205 cells in the order of DATS < DADS < DAS. DATS is the highest for inhibition of migration and invasion of colo 205 cells. DAS, DADS, and DATS induce downregulation expression of PI3K, Ras, MEKK3, MKK7, ERK1/2, JNK1/2, and p38 and then lead to the inhibition of MMP-2, -7, and -9. DAS, DADS, and DATS inhibited NF-κB and COX-2 for leading to the inhibition of cell proliferation. Taken together, these results demonstrated that application of DAS, DADS, and DATS might serve as potential antimetastatic drugs.

Neuroglobin is a highly conserved hemoprotein of uncertain physiological function that evolved from a common ancestor to hemoglobin and myoglobin. It possesses a six-coordinate heme geometry with proximal and distal histidines directly bound to the heme iron, although coordination of the sixth ligand is reversible. We show that deoxygenated human neuroglobin reacts with nitrite to form nitric oxide (NO). This reaction is regulated by redox-sensitive surface thiols, cysteine 55 and 46, which regulate the fraction of the five-coordinated heme, nitrite binding, and NO formation. Replacement of the distal histidine by leucine or glutamine leads to a stable five-coordinated geometry; these neuroglobin mutants reduce nitrite to NO ∼2000 times faster than the wild type, whereas mutation of either Cys-55 or Cys-46 to alanine stabilizes the six-coordinate structure and slows the reaction. Using lentivirus expression systems, we show that the nitrite reductase activity of neuroglobin inhibits cellular respiration via NO binding to cytochrome c oxidase and confirm that the six-to-five-coordinate status of neuroglobin regulates intracellular hypoxic NO-signaling pathways. These studies suggest that neuroglobin may function as a physiological oxidative stress sensor and a post-translationally redox-regulated nitrite reductase that generates NO under six-to-five-coordinate heme pocket control. We hypothesize that the six-coordinate heme globin superfamily may subserve a function as primordial hypoxic and redox-regulated NO-signaling proteins.

A new ruthenium complex, [Ru(L)(bpy) 2 ]·2PF 6 , (L = 3,5-di(2'-pyridyl)-1,2,4-oxadiazole), was synthesized and characterized. The assignment of chemical shift for [Ru(L)(bpy) 2 ]·2PF 6 was accomplished by HHCOSY technique. The cyclic voltammogram of [Ru(L)(bpy) 2 ]·2PF 6 shows a metal-centered Ru II /Ru III oxidation with potential at 1.35 V, as well as four reversible reduction waves with the former two potentials at -1.24 and -1.60 V, attributed to two bpy/bpy - process, and the latter two potentials at -1.86 and -2.16 V, attributed to L/L - and L - /L 2- , respectively. The absorption spectrum of the ruthenium complex was dominated by metal-to-ligand charge transfer (MLCT) band in the visible range, with absorption maxima appearing at 442 nm and molar extinction coefficient of 13 x 10 3 M -1 cm -1 . The phosphorescent emission band has been attributed to from MLCT triplet state.

A family of acyclic aza-bridged bis-1,10-phenanthroline compounds has been synthesized in a convenient way. The resulting compounds 2 and 2·HCl were fully characterized and their solid state structures and NMR spectroscopic properties were investigated to assess how the structural units affect the alkylation reactions. The results reveal the transoid structure for 2. The broadening NMR peak in 2 is shown to be due to an unusual intramolecular CH⋯N hydrogen bond. This unique conformation offers an efficient and regioselective method to prepare the amino-substituted bis-2,2′-1,10-phenanthroline derivatives and 1,10-phenanthrolino-N-alkylated compounds.

Background With respect to interleukin (IL)-1β concentration, the disease-modifying effects of glucosamine HCl (GLN) on IL-1β-treated chondrocytes followed a dose-dependent manner and were traditionally conducted under normoxic conditions not generally encountered by chondrocytes in vivo. Purposes To demonstrate the beneficial effects of GLN on IL-1β-induced matrix metalloproteinase (MMP) expression in chondrosarcoma cells in a dose-dependent manner, but at different concentration ranges of IL-1β; additionally, to determine how hypoxia affects GLN's beneficial effects after dose-dependent application. Methods Under normoxic and hypoxic conditions, human chondrosarcoma cells (SW1353) induced by high and low IL-1β concentrations were respectively treated with low and high concentration ranges of GLN. mRNA and protein expression for MMPs and glucose transporter protein (GLUT)-1 were examined by reverse transcription–polymerase chain reaction and Western blot analysis. Results High concentrations of GLN could reduce mRNA and protein expressions of MMPs induced by low concentration of IL-1β; whereas low concentrations of GLN could reduce the inflammatory response upregulated by high concentrations of IL-1β. The data might substantiate the beneficial effects in clinical application of lower plasma concentrations of GLN achievable by oral administration. Furthermore, the effects were compared under hypoxic conditions to mimic real conditions encountered in human cartilage under normoxia. Upregulation of HIF-1α protein levels under hypoxia was observed. The level of MMP protein expression induced by IL-1β was lower under hypoxic conditions. Instead, inhibition of MMPs by GLN appeared to be more apparent. Finally, GLUT-1 protein expression of SW1353 was upregulated under IL-1β and hypoxia treatment. Conclusion Glucose transporter, most likely GLUT-1, inducible by HIF-1α, might play an important role in the therapeutic efficacy of a lower plasma concentration of GLN on osteoarthritis achievable by oral administration.

Protein engineering strategies seek to develop a hemoglobin-based oxygen carrier with optimized functional properties, including (i) an appropriate O 2 affinity, (ii) high cooperativity, (iii) limited NO reactivity, and (iv) a diminished rate of auto-oxidation. The mutations alphaL29F, alphaL29W, alphaV96W and betaN108K individually impart some of these traits and in combinations produce hemoglobin molecules with interesting ligand-binding and allosteric properties. Studies of the ligand-binding properties and solution structures of single and multiple mutants have been performed. The aromatic side chains placed in the distal-heme pocket environment affect the intrinsic ligand-binding properties of the mutated subunit itself, beyond what can be explained by allostery, and these changes are accompanied by local structural perturbations. In contrast, hemoglobins with mutations in the alpha 1beta 1 and alpha 1beta 2 interfaces display functional properties of both "R"- and "T"-state tetramers because the equilibrium between quaternary states is altered. These mutations are accompanied by global structural perturbations, suggesting an indirect, allostery-driven cause for their effects. Combinations of the distal-heme pocket and interfacial mutations exhibit additive effects in both structural and functional properties, contribute to our understanding of allostery, and advance protein-engineering methods for manipulating the O 2 binding properties of the hemoglobin molecule.

The objective of this study was to evaluate the hepatoprotective activity against the ethanol-induced cytotoxicity by Antrodia cinnamomea which were produced from a large-scale fermentation. The production of biomass, extracellular polysaccharide (EPS), intracellular polysaccharide (IPS), and triterpenoids was 27.1, 1.2, 0.7, and 2.0 g/L, respectively, using a 5000-L agitated bioreactor with 3500 L of A. cinnamomea medium (ACM). The hepatoprotective effects of the water extract from the mycelia of A. cinnamomea (WAC) were evaluated in vitro using ethanol-induced cytotoxicity on AML12 hepatocytes. The cytotoxicity and the apoptosis-associated phosphatidyl serine redistribution of plasma membrane to AML12 cells induced by 300 mM ethanol were effectively reduced by adding 500 mg/L of WAC. From the compositional analysis, the major component in WAC was polysaccharides that showed a high galactose content (70.9 mg/g crude polysaccharide). Hence, the large-scale fermentation was efficient in producing cell mass and its metabolites which were able to protect hepatocytes from the damage by ethanol.

Studies were designed to investigate the effects of baicalein on mouse-rat hybrid retina ganglion cells (N18) to better understand its effect on apoptosis and apoptosis-related genes in vitro. Cell viability, reactive oxygen species (ROS), cytoplasmic Ca2+, mitochondrial membrane potential (MMP), apoptosis induction, and caspases-3 activity were examined by flow cytometric assay. Apoptosis-associated proteins such as p53, Bax, Bcl-2, cytochrome c, and caspase-3 were examined by Western blot. We demonstrated the increase in the levels of p53, Bax, and cytochrome c and decrease in the level of Bcl-2, which are associated with the induction of apoptotic cell death after 24 h treatment with baicalein in N18 cells. Baicalein induced an increase in the cytoplasmic levels of ROS and Ca2+ in 1 h and reached their peak at 3 h, and thereafter a loss of MMP by flow cytometry. We also demonstrated a release of the cytochrome c from mitochondria into cytosol and an activation of caspase-3, which led to the occurrence of apoptosis in N18 cells treated with baicalein by Western blot. Pretreatment was conducted with BAPTA (intracellular calcium chelator) in baicalein-treated cells, the decline of MMP was recovered, and the increase in the level of cytoplasmic Ca2+ was suppressed, and the proportion of apoptosis was also markedly diminished. In conclusion, our data suggests that oxidative stress and cellular Ca2+ modulates the baicalein-induced cell death via a Ca2+-dependent mitochondrial death pathway in N18 cells.

Taiwanofungus camphoratus (T. camphoratus), a fungus and a Taiwan-specific, well-known traditional Chinese medicine, has long been used to treat diarrhea, hypertension, itchy skin, and liver cancer. To gain a large amount of T. camphoratus, several culture techniques have been developed, including solid-state culture and liquid-state fermentation. Peroxisome proliferator-activated receptor gamma (PPARgamma) has been described as a hypoglycemic agent that increases insulin sensitivity in peripheral tissues and results in reduced blood glucose, insulin, and triglyceride levels in insulin-resistant animals and in type-2 (non-insulin-dependent) diabetic patients. In this study, we investigate the possibility that T. camphoratus might activate PPARgamma in vitro and hypolipidemic activity in vivo. The results show that an aqueous extract of the wild fruiting bodies of T. camphoratus was able to increase the PPARgamma activity in cells transfected with the PPARgamma expression plasmid and the AOx-TK reporter plasmid. Based on the cell experiment, we examined the hypolipidemic effect of wild fruiting bodies (WFT) and a solid-state culture (SST) of T. camphoratus on SD rats fed on a high-cholesterol (HC) diet. The results show that WFT significantly decreased the serum triglyceride level, but could not affect the cholesterol level. SST only slightly decreased the serum triglyceride level. In addition, both WFT and SST significantly decreased the serum alanine transaminase (ALT) level and protected against the liver damage induced by the HC diet from the results of a histological examination. These results suggest that T. camphoratus might contain PPARgamma ligands and result in a hypotriglyceridemic effect, and that it also exhibits a liver protective activity.

The purpose of this study was to investigate the possible involvement of synovium in cartilage destruction in osteoarthritis (OA) patients. Using human primary synovial fibroblasts (HPSFs), we examined the effects of glucosamine (GLN) on the regulation of the expression of matrix metalloproteinases (MMP-1, -2, and -13) and chemokines (IL-8, MCP-1, and RANTES) as well as the involvement of MAPK signal pathways (JNK, ERK, and p-38) and the transcription factor of NF-kappaB on the present or absence of interleukin (IL)-1beta. Our experiments showed that protein production and mRNA expressions of MMP-1, MMP-3, MMP-13, IL-8, MCP-1, and RANTES were downregulated by treatment with glucosamine in HPSFs. The results further showed that GLN could inhibit IkappaBalpha phosphorylation and IkappaBalpha degradation leading to inhibition of the translocation of NF-kappaB to nuclei. However, GLN upregulated MAPKs pathways in HPSFs cells with or without IL-1beta. The results suggest that the inhibition of MMP-1, -3, and -13 expressions as well as IL-8, MCP-1, and RANTES productions by GLN might mediate suppression of NF-kappaB signal pathways, and HPSFs seem to have a potential functions as an alternative source of MMPs and chemokines for inducing the degradation of cartilage in OA.

The retention behaviors and efficiencies of the chromatographic separation of dextromethorphan (DM) and its metabolites (dextrorphan (DX), 3‐methoxymorphinan (MM), and 3‐hydroxymorphinan (HM)), on three different silica columns (Inertsil, μ‐Porasil, and Lichrospher), were compared using a simple aqueous mobile phase consisting of an organic solvent (methanol or acetonitrile) and water at different volume ratios (1∶9 to 9∶1) containing triethylamine (TEA) and acetic acid (ACH). Results demonstrated that the retention capacities for basic compounds with the same ionization conditions of the silanol group were the largest for the Lichrospher column, followed by the μ‐Porasil one, with the Inertsil column exhibiting the lowest level. Based on their physical characteristics, the larger retention capacities for DM and its three metabolites in the LiChrospher column compared to those of the Inertsil and μ‐Porasil columns may have been because the LiChrospher column has the largest surface area for inter...

Phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P(2)) has been known to serve as a substrate for phosphatidylinositol 3-kinase (PI(3)K) and phosphoinositide-specific phospholipase C (PI-PLC), which can produce PtdIns(3,4,5)P(3) and inositol 1,4,5-trisphosphate (Ins(1,4,5)P(3)) and diacylglycerol (DAG), respectively. In this study, we elucidated the role of PI-PLC during the LPS-activated mouse macrophages RAW264.7 treated with PI(3)K inhibitor wortmannin. First, wortmannin treatment enhanced Ins(1,4,5)P(3) production and iNOS expression in LPS-activated macrophages. Inhibition of PI(3)K by p85 siRNA also showed an enhancement of iNOS expression. On the other hand, overexpression of PI(3)K by ras-p110 expression plasmid significantly decreased iNOS expression in LPS-activated macrophages. In addition, overexpression of wild-type or dominant-negative Akt expression plasmid did not affect the iNOS expression in LPS-activated macrophages. Second, treatment of PI-PLC inhibitor U73122 reversed the enhancement of iNOS expression, the increase of phosphorylation level of ERK, JNK and p38, and the increase of AP-1-dependent gene expression in wortmannin-treated and LPS-activated macrophages. However, NF-kappaB activity determined by EMSA assay and reporter plasmid assay did not change during LPS-activated macrophages with or without wortmannin. We propose that the inhibition of PI(3)K by wortmannin in mouse macrophages enhances the PI-PLC downstream signals, and subsequently increases the LPS induction of iNOS expression independently of Akt pathway.

In attempting to perturb the electronic properties of the spacer, we now describe an interesting example of Ru 2+ –tpy (tpy = terpyridine) complexes with 1,1 0 -bis(ethynyl)polyferrocenyl moiety attached directly to the 4 0 -position of the tpy ligand (tpy–C„C–(fc)n–C„C–tpy; fc = ferrocenyl; n = 2–3). Complexes of Ru 2+ –tpy have room-temperature luminescence in H2O/CH3CN (4/1) solution. The ground-state HOMO and LUMO energies were probed by electrochemical measurements and the excited-state photophysical properties were probed by UV–Vis absorption spectroscopy and luminescence spectroscopy. The redox behavior of [(tpy)Ru II –tpy–C„C–(fc)n–C„C–tpy– Ru II (tpy)] 4+ complex is dominated by the Ru 2+ /Ru 3+ redox couple (E1/2 from 1.35 to 1.39 V), Fe 2+ /Fe 3+ redox couples (E1/2 from 0.4 to 1.0 V) and tpy/tpy � /tpy 2� redox couples (E1/2 from � 1.3 to � 1.5 V). Electrochemical data, UV absorption and emission spectra indicate that the p-delocalization in the spacer is enhanced by the insertion of ethynyl unit. Interestingly, the insertion of ethynyl unit into the main chain causes a dramatic increase of phosphorescence yield (1.48 · 10 � 4 for n = 2; 1.13 · 10 � 4 for n = 3), triplet lifetime (67 ns for n =2 ; 24 ns for n = 3), and emission intensity. The biferrocenyl spacer can be converted into mixed-valence biferrocenium spacer, which gives a more effectivep-delocalization along main chain, by selective chemical oxidation of ferrocenyl unit. In deoxygenated H2O/CH3CN (4/1) solution at 25 � C, the oxidized complex of [(tpy)Ru II –tpy–C„C–(fc)2–C„C–tpy–Ru II (tpy)] 5+ is nonemissive. The presence of lower energy ferroce

Background Osteoarthritis (OA) is a common joint disease that causes disabilities in elderly. However, few agents with high efficacy and low side effects have been developed to treat OA. In this study, we evaluated the effects of the alginate extract named CTX in OA cell and rabbit models. Results CTX was formulated by hydrolyzing sodium alginate polymers with alginate lyase and then mixing with pectin. HPLC was used to analyze the CTX content. Human chondrosarcoma SW1353 cells treated with interleukin-1β were used as OA model cells to investigate the effects of CTX on chondrocyte inflammation and anabolism. CTX at concentrations up to 1000 μg/ml exerted low cytotoxicity. It inhibited the gene expression of proinflammatory matrix metalloproteinases (MMPs) including MMP1, MMP3 and MMP13 in a dose-dependent manner and increased the mRNA level of aggrecan, the major proteoglycan in articular cartilage, at 1000 μg/ml. Thirteen-week-old New Zealand White rabbits underwent a surgical anterior cruciate ligament transection and were orally treated with normal saline, glucosamine or CTX for up to 7 weeks. Examinations of the rabbit femur and tibia samples demonstrated that the rabbits taking oral CTX at a dosage of 30 mg/kg/day suffered lesser degrees of articular stiffness and histological cartilage damage than the control rabbits. Conclusions The gene expression profiles in the cell and the examinations done on the rabbit cartilage suggest that the alginate extract CTX is a pharmaco-therapeutic agent applicable for OA therapy.

The aim of this study was to purify protein(s) from Piper betle leaf for identification and further characterization. A functionally unknown protein was purified to apparent homogeneity with a molecular mass of 15.7 kDa and identified as Cu/Zn superoxide dismutase (SOD). The purified SOD appeared to be monomeric and converted to its dimeric form with increased enzymatic activity in betel nut oral extract. This irreversible conversion was mainly induced by slaked lime, resulting from the increase in pH of the oral cavity. Oral extract from chewing areca nut alone also induced SOD dimerization due to the presence of arginine. The enhanced activity of the SOD dimer was responsible for the continuous production of hydrogen peroxide in the oral cavity. Thus, SOD may contribute to oral carcinogenesis through the continuous formation of hydrogen peroxide in the oral cavity, in spite of its protective role against cancer in vivo.

One cuprous complex [Cu(L)(PPh 3 )I] was obtained by reaction of [Cu(PPh 3 ) 3 I] (PPh 3 = triphenylphosphine) with a new α-diimine ligand L (N-diphenylmethylene-3,5-di(2'-pyridyl)-4-amino-1,2,4-triazole). The complex has been characterized by UV-Vis, luminescence, 1 H NMR spectroscopy and cyclic voltammetric study. The structure of ligand L and complex are confirmed by single crystal X-ray diffraction study. The compound [Cu(L)(PPh 3 )I] crystallizes in the monoclinic P21/c space group with a = 19.9291(4), b = 10.0691(2), c =21.9245(5), a = y = 90, β = 92.481(10), and Z = 4. The complex shows a MLCT band in the 350∼545 nm region and exhibits luminescent emission at 600 nm at room temperature in dichloromethane solution. Cyclic voltammogram of [Cu(L)(PPh 3 )I] shows a quasi-reversible redox couple of the copper center at 0.67 V for Cu(I)/Cu(II) and an irreversible reduction potential at -0.98 V accompanied by a stripping peak of copper at -0.11 V for Cu(I)/Cu(0).

Pectin is composed of complex polysaccharides rich in galactoside residues, and it is most abundant in citrus fruits. Pectin and modified pectin have been found to exhibit anti-mutagenic activity and inhibit cancer metastasis and proliferation, with no evidence of toxicity or other serious side effects. In this study, we investigated the inhibitory effect of pectin at different degrees of esterification (DEs) on the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated macrophages. Western blot and RT-PCR analyses demonstrated that 30% esterified pectin (DE30), DE60 pectin, and DE90 pectin significantly inhibited the protein and mRNA expressions of iNOS and COX-2 in LPS-activated macrophages, and DE90 pectin was the most-potent inhibitor. To clarify the mechanisms involved, DE90 pectin was found to inhibit the phosphorylation of MAPKs and IKK kinase activity. In addition, DE90 pectin inhibited the activation of NF-kappaB and AP-1 by electrophoretic mobility shift assay and transient transfection experiments. Finally, we found that DE90 pectin could bind with LPS, and might result in decreased binding of LPS to its receptor. These results suggest that modulation of iNOS and COX-2 expressions by dietary pectin may be important in cancer chemoprevention and anti-inflammation.

Two cuprous complexes Cu(L1)(PPh3)I (1) and Cu(L2)(PPh3)I (2) were obtained by reaction of Cu(PPh3)3I (PPh3 = triphenylphosphine) with two different α-diimine ligands L 1 (3,5-di(2′-pyridyl)-4-amin...

Terbinafine (TB, lamisil ® ), a promising world widely used oral-anti-fungal agent, has been used in the treatment of superficial mycosis. In this study, we found that apoptosis but not cell growth arrest was induced by TB (1 μM, for 24 h) in human promyelocytic leukemia (HL60) cells. The apoptotic effect induced by TB in the HL60 cell was not through the general differentiation mechanisms evidenced by evaluation of three recognized markers, including CD11b, CD33, and morphological features. In addition, our results also revealed that TB-induced apoptosis was not through the cellular surface CD 95 receptor-mediated signaling pathway. We found that the mitochondria membrane in the TB-treated HL60 cells was dissipated by decreasing of the electrochemical gradient (Δ Ψ m ) led to leakage of cytochrome c from mitochondria into cytosol. Such effects were completely blocked by in vitro transfection of the HL60 cells with Bcl-2 overexpression plasmid (HL60/Bcl-2). However, our data found that TB-mediated apoptosis could not be completely prevented in the Bcl-2 over expressed (HL60/Bcl-2) cells. Such results implied that additional mediators (such as caspase-9) other than mitochondria membrane permeability might contribute to the TB-induced cellular apoptosis signaling. This hypothesis was supported by the evidence that administration of caspases-9 specific inhibitor (z-LEHD-fmk) blocked the TB-induced apoptosis. Our studies highlight the molecular mechanisms of TB-induced apoptosis in human promyelocytic leukemia (HL60) cells.