Search results for: mechanochemical activation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 968

Search results for: mechanochemical activation

788 Curcumin Attenuates Angiogenesis in Liver Fibrosis and Inhibits Angiogenic Properties of Hepatic Stellate Cells

Authors: Feng Zhang, Li Chen, Desong Kong, Xiaoping Zhang, Xiaojing Zhu, Yin Lu, Shizhong Zheng

Abstract:

Sinusoidal pathological angiogenesis is a novel therapeutic target for liver fibrosis. We demonstrated that curcumin ameliorated fibrotic injury and sinusoidal angiogenesis in rat liver with fibrosis caused by carbon tetrachloride. Curcumin reduced the expression of angiogenic markers in fibrotic liver. Experiments in vitro showed that the viability and vascularization of rat liver sinusoidal endothelial cells (LSECs) were not impaired by curcumin. Further investigations showed that curcumin inhibited VEGF expression in hepatic stellate cells (HSCs) by disrupting PDGF-βR/ERK and mTOR pathways. HSC motility and vascularization were also suppressed by curcumin via blocking PDGF-βR/FAK/RhoA cascade. Gain- or loss-of-function analyses revealed that activation of PPARγ was required for curcumin to inhibit angiogenic properties of HSCs. We concluded that curcumin attenuated sinusoidal angiogenesis in liver fibrosis possibly by targeting HSCs via a PPARγ activation-dependent mechanism. PPARγ could be a target molecule for reducing pathological angiogenesis during liver fibrosis.

Keywords: angiogenesis, hepatic stellate cell, curcumin, peroxisome proliferator-activated receptor-γ

Procedia PDF Downloads 474
787 Atmospheric Oxidation of Carbonyls: Insight to Mechanism, Kinetic and Thermodynamic Parameters

Authors: Olumayede Emmanuel Gbenga, Adeniyi Azeez Adebayo

Abstract:

Carbonyls are the first-generation products from tropospheric degradation reactions of volatile organic compounds (VOCs). This computational study examined the mechanism of removal of carbonyls from the atmosphere via hydroxyl radical. The kinetics of the reactions were computed from the activation energy (using enthalpy (ΔH**) and Gibbs free energy (ΔG**). The minimum energy path (MEP) analysis reveals that in all the molecules, the products have more stable energy than the reactants, which implies that the forward reaction is more thermodynamically favorable. The hydrogen abstraction of the aromatic aldehyde, especially without methyl substituents, is more kinetically favorable compared with the other aldehydes in the order of aromatic (without methyl or meta methyl) > alkene (short chain) > diene > long-chain aldehydes. The activation energy is much lower for the forward reaction than the backward, indicating that the forward reactions are more kinetically stable than their backward reaction. In terms of thermodynamic stability, the aromatic compounds are found to be less favorable in comparison to the aliphatic. The study concludes that the chemistry of the carbonyl bond of the aldehyde changed significantly from the reactants to the products.

Keywords: atmospheric carbonyls, oxidation, mechanism, kinetic, thermodynamic

Procedia PDF Downloads 11
786 Synthesis of 2-Aminoisocoumarinoselenazoles via Transition Metal-Free Alkylation and Ru(II)-Catalyzed [4+2] Alkyne Annulation

Authors: Sunil Kumar, Sandip Dhole, Deepak Salunke, Chung-ming Sun

Abstract:

Heterocycles bearing nitrogen, oxygen, and selenium are present in innumerable biologically active compounds. For instance, coumarin containing dicoumarol acts as naturally occurring anticoagulant. 2-Acylamido selenazole works as Store-Operated Calcium (SOC) channel regulator. Therefore, due to biologically significance of selenazole and coumarin and our quest to develop efficient methodologies for the synthesis of complex heterocycles, the trisubstituted angular isocoumarinoselenazole synthesis was proposed and achieved by starting from nitrobenzoic acid derivative, available commercially. Synthetic procedure involves three steps: i) the construction of 2-aminobenzoselenazoles, ii) their regioselective N-alkylation at position-2 and iii) alkyne insertion via Ru catalyzed C-H activation. Transition metal free synthesis of benzoselenazoles was successfully brought about by the addition/elimination reaction via intramolecular C-Se bond formation. In the next step, N-alkylation of selenazole furnished two regioisomers. Both the isomers exhibited different reactivity towards [4+2] alkyne annulation reaction. The fusion of α-pyrone ring on the benzo[1,3-d]selenazole skeleton was achieved via Ru(II)-catalyzed C-H activation and alkyne insertion. As evident from mechanism, the selenazole 'N' plays an important role for the experiential selectivity.

Keywords: alkylation, alkyne insertion, coumarin, selenazole

Procedia PDF Downloads 92
785 Correlation Mapping for Measuring Platelet Adhesion

Authors: Eunseop Yeom

Abstract:

Platelets can be activated by the surrounding blood flows where a blood vessel is narrowed as a result of atherosclerosis. Numerous studies have been conducted to identify the relation between platelets activation and thrombus formation. To measure platelet adhesion, this study proposes an image analysis technique. Blood samples are delivered in the microfluidic channel, and then platelets are activated by a stenotic micro-channel with 90% severity. By applying proposed correlation mapping, which visualizes decorrelation of the streaming blood flow, the area of adhered platelets (APlatelet) was estimated without labeling platelets. In order to evaluate the performance of correlation mapping on the detection of platelet adhesion, the effect of tile size was investigated by calculating 2D correlation coefficients with binary images obtained by manual labeling and the correlation mapping method with different sizes of the square tile ranging from 3 to 50 pixels. The maximum 2D correlation coefficient is observed with the optimum tile size of 5×5 pixels. As the area of the platelet adhesion increases, the platelets plug the channel and there is only a small amount of blood flows. This image analysis could provide new insights for better understanding of the interactions between platelet aggregation and blood flows in various physiological conditions.

Keywords: platelet activation, correlation coefficient, image analysis, shear rate

Procedia PDF Downloads 304
784 Phorbol 12-Myristate 13-Acetate (PMA)-Differentiated THP-1 Monocytes as a Validated Microglial-Like Model in Vitro

Authors: Amelia J. McFarland, Andrew K. Davey, Shailendra Anoopkumar-Dukie

Abstract:

Microglia are the resident macrophage population of the central nervous system (CNS), contributing to both innate and adaptive immune response, and brain homeostasis. Activation of microglia occurs in response to a multitude of pathogenic stimuli in their microenvironment; this induces morphological and functional changes, resulting in a state of acute neuroinflammation which facilitates injury resolution. Adequate microglial function is essential for the health of the neuroparenchyma, with microglial dysfunction implicated in numerous CNS pathologies. Given the critical role that these macrophage-derived cells play in CNS homeostasis, there is a high demand for microglial models suitable for use in neuroscience research. The isolation of primary human microglia, however, is both difficult and costly, with microglial activation an unwanted but inevitable result of the extraction process. Consequently, there is a need for the development of alternative experimental models which exhibit morphological, biochemical and functional characteristics of human microglia without the difficulties associated with primary cell lines. In this study, our aim was to evaluate whether THP-1 human peripheral blood monocytes would display microglial-like qualities following an induced differentiation, and, therefore, be suitable for use as surrogate microglia. To achieve this aim, THP-1 human peripheral blood monocytes from acute monocytic leukaemia were differentiated with a range of phorbol 12-myristate 13-acetate (PMA) concentrations (50-200 nM) using two different protocols: a 5-day continuous PMA exposure or a 3-day continuous PMA exposure followed by a 5-day rest in normal media. In each protocol and at each PMA concentration, microglial-like cell morphology was assessed through crystal violet staining and the presence of CD-14 microglial / macrophage cell surface marker. Lipopolysaccharide (LPS) from Escherichia coli (055: B5) was then added at a range of concentrations from 0-10 mcg/mL to activate the PMA-differentiated THP-1 cells. Functional microglial-like behavior was evaluated by quantifying the release of prostaglandin (PG)-E2 and pro-inflammatory cytokines interleukin (IL)-1β and tumour necrosis factor (TNF)-α using mediator-specific ELISAs. Furthermore, production of global reactive oxygen species (ROS) and nitric oxide (NO) were determined fluorometrically using dichlorodihydrofluorescein diacetate (DCFH-DA) and diaminofluorescein diacetate (DAF-2-DA) respectively. Following PMA-treatment, it was observed both differentiation protocols resulted in cells displaying distinct microglial morphology from 10 nM PMA. Activation of differentiated cells using LPS significantly augmented IL-1β, TNF-α and PGE2 release at all LPS concentrations under both differentiation protocols. Similarly, a significant increase in DCFH-DA and DAF-2-DA fluorescence was observed, indicative of increases in ROS and NO production. For all endpoints, the 5-day continuous PMA treatment protocol yielded significantly higher mediator levels than the 3-day treatment and 5-day rest protocol. Our data, therefore, suggests that the differentiation of THP-1 human monocyte cells with PMA yields a homogenous microglial-like population which, following stimulation with LPS, undergo activation to release a range of pro-inflammatory mediators associated with microglial activation. Thus, the use of PMA-differentiated THP-1 cells represents a suitable microglial model for in vitro research.

Keywords: differentiation, lipopolysaccharide, microglia, monocyte, neuroscience, THP-1

Procedia PDF Downloads 349
783 Fabrication of Activated Carbon from Palm Trunksfor Removal of Harmful Dyes

Authors: Eman Alzahrani

Abstract:

Date palm trees are abundant and cheap natural resources in Saudi Arabia. In this study, an activated carbon was prepared from palm trunks by chemical processes. The chemical activation was performed by impregnation of the raw materials after grinding with H3PO4 solution (63%), followed by placing of the sample solution on a muffle furnace at 400ºC for 30 min, and then at 800ºC for 10 min. The morphology of the fabricated material was checked using scanning electron microscopy that showed the rough surfaces on the carbon samples. The use of fabricated activated carbon for removal of eosin dye from aqueous solutions at different contact time, initial dye concentration, pH and adsorbent doses was investigated. The experimental results show that the adsorption process attains equilibrium within 20 min. The adsorption isotherm equilibrium was studied by means of the Langmuir and Freundlich isotherms, and it was found that the data fit the Langmuir isotherm equation with maximum monolayer adsorption capacity of 126.58 mg g-1. The results indicated that the home made activated carbon prepared from palm trunks has the ability to remove eosin dye from aqueous solution and it will be a promising adsorbent for the removal of harmful dyes from waste water.

Keywords: activated carbon, date palm trunks, H3PO4 activation, adsorption, dye removal, eosin dye, isotherm

Procedia PDF Downloads 327
782 Development of 3D Neck Muscle to Analyze the Effect of Active Muscle Contraction in Whiplash Injury

Authors: Nisha Nandlal Sharma, Julaluk Carmai, Saiprasit Koetniyom, Bernd Markert

Abstract:

Whiplash Injuries are mostly experienced in car accidents. Symptoms of whiplash are commonly reported in studies, neck pain and headaches are two most common symptoms observed. The whiplash Injury mechanism is poorly understood. In present study, hybrid neck muscle model were developed with a combination of solid tetrahedral elements and 1D beam elements. Solid tetrahedral elements represents passive part of the muscle whereas, 1D beam elements represents active part. To simulate the active behavior of the muscle, Hill-type muscle model was applied to beam elements. To simulate non-linear passive properties of muscle, solid elements were modeled with rubber/foam material model. Some important muscles were then inserted into THUMS (Total Human Model for Safety) THUMS was given a boundary conditions similar to experimental tests. The model was exposed to 4g and 7g rear impacts as these load impacts are close to low speed impacts causing whiplash. The effect of muscle activation level on occupant kinematics during whiplash was analyzed.

Keywords: finite element model, muscle activation, THUMS, whiplash injury mechanism

Procedia PDF Downloads 302
781 Polyimide Supported Membrane Made of 2D-Coordination-Crosslinked Polyimide for Rapid Molecular Separation in Multi-Solvent Environments

Authors: Netsanet Kebede Hundessa

Abstract:

Substrate modification of thin film composite (TFC) membranes with various crosslinkers is typically necessary for organic solvent nanofiltration (OSN) applications. This modification is aimed at enhancing membrane stability and solvent resistance, but it often results in a decline in permeance. This study introduces a distinct approach by developing a coordination-crosslinked polyimide substrate, which differs from the covalently-crosslinked substrates traditionally used. This developed substrate achieves enhanced solvent resistance, improved hydrophilicity, and optimized porous microstructure simultaneously. The study investigates the effects of an alkaline coagulation bath, subsequent ion exchange, and further solvent activation. The resulting TFC membrane successfully overcomes the typical permeability-selectivity trade-off of OSN membranes. It demonstrates significantly improved solvent permeance (1.5–2 times higher than previously reported data) with values of 65.2 LMH/bar for methanol, 33.1 LMH/bar for ethanol, and 59.1 LMH/bar for acetone while maintaining competitive solute rejection (>98% for Rose Bengal). This research is expected to provide a new direction for developing high-performance OSN composite membranes and other separation applications.

Keywords: metal coordinatiom, thin film composite membrane, organic solvent nanofiltration, solvent activation

Procedia PDF Downloads 19
780 Arsenic and Mercury Levels in Scalp Hair of School Children of Three Villages in Kandal Province, Cambodia

Authors: Alireza Yavar, Sukiman Sarmani, Khoo Kok Siong

Abstract:

The residents of villages in Kandal province of Cambodia, because of dietary habits, lifestyle and ecological conditions, are exposed to toxic elements like arsenic (As) and mercury (Hg). For comparison purpose, scalp hair samples of 12-17 school children from three villages of Anglong Romiot (AR), Svay Romiot (SR) and Kampong Kong (KK) in Kandal province of Cambodia were considered using k0- instrumental neutron activation method (k0-INAA). The samples irradiated 6 hours with 750 kW power in Malaysian nuclear agency (MNA) research reactor and subsequently found gamma peaks of radionuclides in samples using HPGe detector. The average values of arsenic and mercury were 0.0 and 3.52 (mg/kg) in AR; 1.88 and 4.26 (mg/kg) in SR; 2.81 and 3.37 (mg/kg) in KK, respectively. The results indicate KK, SR, and AR villages were in high, medium and control level of arsenic pollution, respectively. However, Hg concentration were highest in SR, then KK and AR villages, respectively. The accuracy of the method was assessed by analyzing ERM-DB001-human hair as certified reference materials (CRMs), which experimental result of ERM-DB001 was consistent with certified values. In addition, correlation between As and Hg levels was found by Pearson’s correlation test.

Keywords: Kandal province of Cambodia, k0- instrumental neutron activation method., scalp human hair, arsenic and mercury

Procedia PDF Downloads 60
779 Study of Functional Relevant Conformational Mobility of β-2 Adrenoreceptor by Means of Molecular Dynamics Simulation

Authors: G. V. Novikov, V. S. Sivozhelezov, S. S. Kolesnikov, K. V. Shaitan

Abstract:

The study reports about the influence of binding of orthosteric ligands as well as point mutations on the conformational dynamics of β-2-adrenoreceptor. Using molecular dynamics simulation we found that there was a little fraction of active states of the receptor in its apo (ligand free) ensemble corresponded to its constitutive activity. Analysis of MD trajectories indicated that such spontaneous activation of the receptor is accompanied by the motion in intracellular part of its alpha-helices. Thus receptor’s constitutive activity directly results from its conformational dynamics. On the other hand the binding of a full agonist resulted in a significant shift of the initial equilibrium towards its active state. Finally, the binding of the inverse agonist stabilized the receptor in its inactive state. It is likely that the binding of inverse agonists might be a universal way of constitutive activity inhibition in vivo. Our results indicate that ligand binding redistribute pre-existing conformational degrees of freedom (in accordance to the Monod-Wyman-Changeux-Model) of the receptor rather than cause induced fit in it. Therefore, the ensemble of biologically relevant receptor conformations is encoded in its spatial structure, and individual conformations from that ensemble might be used by the cell in conformity with the physiological behaviour.

Keywords: seven-transmembrane receptors, constitutive activity, activation, x-ray crystallography, principal component analysis, molecular dynamics simulation

Procedia PDF Downloads 222
778 Effects of β-Glucan on the Release of Nitric Oxide by RAW264.7 Cells Stimulated with Escherichia coli Lipopolysaccharide

Authors: Eun Young Choi, So Hui Choe, Jin Yi Hyeon, Ji Young Jin, Bo Ram Keum, Jong Min Lim, Hyung Rae Cho, Kwang Keun Cho, In Soon Choi

Abstract:

This research analyzed the effect of β-glucan that is expected to alleviate the production of inflammatory mediator in macrophagocyte, which was processed by the lipopolysaccharide (LPS) of Escherichia, a pathogen related to allergy. The incubated layer was used for nitric oxide (NO) analysis. The DNA-binding activation of the small unit of NF-κB was measured using ELISA-based kit. In RAW264.7 cells that were vitalized by E.coli LPS, β-glucan inhibited both the combatant and rendering phases of inducible NO synthase (iNOS)-derived NO. β-glucan increased the expression of heme oxygenase-1 (HO-1) in the cell that was stimulated by E.coli LPS, and HO-1 activation was inhibited by SnPP. This shows that NO production induced by LPS is related to the inhibition effect of β-glucan. The phosphorylation of JNK and p38 induced by LPS were not influenced by β-glucan, and IκB-α decomposition was not influenced either. Instead, β-glucan remarkably inhibited the phosphorylation of STAT1 that was induced by E.coli LPS. Overall, β-glucan inhibited the production of NO in macrophagocyte that was vitalized by E.coli LPS through HO-1 induction and STAT1 pathways inhibition in this research. As the host inflammation reaction control by β-glucan weakens the progress of allergy, β-glucan can be used as an effective treatment method.

Keywords: β-glucan, lipopolysaccharide (LPS), nitric oxide (NO), RAW264.7 cells, STAT1

Procedia PDF Downloads 371
777 The Negative Impact of Mindfulness on Creativity: An Experimental Test

Authors: Marine Agogue, Beatrice Parguel, Emilie Canet

Abstract:

Defined as receptive attention to and awareness of present events and experience, mindfulness has grown in popularity over the past 30 years to become a trendy buzzword in business media, which regularly reports on its organizational benefits. Mindfulness would enhance or impede creative thinking depending on the type of meditation. Specifically, focused-attention meditation (focusing attention on one object instead of being open to perceive and observe any sensation or thought) would not be or negatively correlated to creativity. This research explores whether mood, in its two dimensions (i.e., hedonic tone, activation level), could mediate this potentially negative effect. The rationale is that focused-attention meditation is likely to improve hedonic tone but, in the meantime, damage activation level, resulting in opposite effects on creativity through the mediation effect of creative self-efficacy, i.e., the belief that one can perform successfully in an ideation setting. To test this conceptual model, a survey was administered to 97 subjects (53% women, mean age: 25 years), randomly assigned to three conditions (a 10-minute focused-attention meditation session vs. a 10-minute psychometric tests session vs. a control condition) and asked to participate in the egg creative task. Creativity was measured in terms of fluency, expansivity, and originality, the other variables using existing scales: hedonic tone (e.g., joyful, happy), activation level (e.g., passive, sluggish), creative self-efficacy (e.g., ‘I felt confident in my ability to do the task effectively’) and self-perceived creativity (e.g., ‘I have lots of original ideas’). The chains of mediation were tested using PROCESS macro (model 6) and controlled for subjects’ gender, age, and self-perceived creativity. Comparing the mindfulness and the control conditions, no difference appeared in terms of creativity, nor any mediation chain by hedonic tone. However, subjects who participated in the meditation session felt less active than those in the control condition, which decreased their creative self-efficacy, and creativity (whatever the indicator considered). Comparing the mindfulness and the psychometric tests conditions, analyses showed that creativity was higher in the psychometric tests condition. As previously, no mediation chain appeared by hedonic tone. However, subjects who participated in the meditation session felt less active than those in the psychometric tests condition, which decreased their creative self-efficacy, and creativity. These findings confirm that focused-attention meditation does not enhance creativity. They demonstrate an emotional underlying mechanism based on activation level and suggest that both positive and active mood states have the potential to enhance creativity through creative self-efficacy. In the end, they should discourage organizations from trying to nudge creativity using mindfulness ad hoc devices.

Keywords: creativity, mindfulness, creative self-efficacy, experiment

Procedia PDF Downloads 98
776 Roles of Lysine-63-Linked Ubiquitination in Cell Decision Fate between Cell Proliferation and Apoptosis

Authors: Chargui Abderrahman, Nehdi Afef , BelaïD Amine , Djerbi Nadir, Tauc Michel, Hofman Paul, Mograbi Baharia, El May MichèLe

Abstract:

K63-linked ubiquitination — i.e. conjugation of a chain of ubiquitins (Ub) linked through lys63 — has emerged as a key mechanism regulating signalling transduction pathways. Although critical, very little information is currently available about how subversion of K63 ubiquitination might contribute to cancers and inflammatory diseases. The present study provides the first evidence that Cadmium (Cd), a widespread environmental carcinogen and toxicant, is a powerful activator of K63 ubiquitination. Indeed, Cd induces accumulation of K63 polyUb proteins. Importantly, Cd-induced ubiquitination does not stem on oxidative damage or proteasome impairment. Rather, we demonstrate that Cd not only activates K63 ubiquitination but also amplifies their accumulation by overloading the capacity of autophagy pathway. At molecular level, Cd-induced ubiquitination is correlated with stabilization of HIF-1 and the activation of NF-B, two transcription factors. Strikingly, prolonged cell exposure to high Cd concentrations induces an exaggerated K63 ubiquitination that fosters aggresome formation, thus precluding these proteins from interacting with their downstream nuclear targets. We therefore propose that the aberrant activation of K63 ubiquitination by the carcinogen Cadmium could promote cell proliferation and inflammation at low levels while high levels committed cell to death.

Keywords: cadmium, environmental exposure, Lysine-63-ubiquitination, kidney, apoptosis, proliferation, autophagy

Procedia PDF Downloads 172
775 Production of Amorphous Boron Powder via Chemical Vapor Deposition (CVD)

Authors: Meltem Bolluk, Ismail Duman

Abstract:

Boron exhibits the properties of high melting temperature (2273K to 2573 K), high hardness (Mohs: 9,5), low density (2,340 g/cm3), high chemical resistance, high strength, and semiconductivity (band gap:1,6-2,1 eV). These superior properties enable to use it in several high-tech areas from electronics to nuclear industry and especially in high temperature metallurgy. Amorphous boron and crystalline boron have different application areas. Amorphous boron powder (directly amorphous and/or α-rhombohedral) is preferred in rocket firing, airbag inflating and in fabrication of superconducting MgB2 wires. The conventional ways to produce elemental boron with a purity of 85 pct to 95 prc are metallothermic reduction, fused salt electrolysis and mechanochemical synthesis; but the only way to produce high-purity boron powders is Chemical Vapour Deposition (Hot Surface CVD). In this study; amorphous boron powders with a minimum purity of 99,9 prc were synthesized in quartz tubes using BCl3-H2 gas mixture by CVD. Process conditions based on temperature and gas flow rate were determined. Thermodynamical interpretation of BCl3-H2 system for different temperatures and molar rates were performed using Fact Sage software. The characterization of powders was examined by using Xray diffraction (XRD), Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM), Stereo Microscope (SM), Helium gas pycnometer analysis. The purities of final products were determined by titration after lime fusion.

Keywords: amorphous boron, CVD, powder production, powder characterization

Procedia PDF Downloads 182
774 An Experimental Approach of the Reuse of Dredged Sediments in a Cement Matrix by Physical and Heat Treatment

Authors: Mahfoud Benzerzour, Mouhamadou Amar, Nor-edine Abriak

Abstract:

In this study, a sediment was used as a secondary raw material in cement substitution with prior treatment. The treatment adopted is a physical treatment involving grinding and separation to obtain different fractions, using a dry method (1 mm, 250µm, 120µm) and washing method (250µm and 120µm). They were subsequently heat treated at temperatures of 650°C, 750°C and 850°C for 1 hour and 3 hours, in order to enable chemical activation by decarbonation or by pozzolanic activation of the material. Different characterization techniques were performed. The determination of main physical and chemical characteristics was obtained through multiple tests: particle size distribution, specific density, the BET surface area, the initial setting time and hydration heat calorimetry Langavant. The chemical tests include: ATG analysis, X-ray diffractometry (XRD) and X-ray fluorescence (XRF) which were used to quantify the fractions, phases and chemical elements present. Compression tests were performed conforming NF EN 196-1 French standard, over terms of 7 days - 14 days - 28 days and 60 days on all formulated mortars: reference mortar based on 100% CEM I 52.5N binder and cement substituted mortars with 8% and 15% by treated sediment. This clearly evidenced contribution due to the chemical activity which was confirmed by calorimetry monitoring and strength investigation.

Keywords: sediment, characterization, grinding, heat treatment, substitution

Procedia PDF Downloads 170
773 Activation of NLRP3 Inflammasomes by Helicobacter pylori Infection in Innate Cellular Model and Its Correlation to IL-1β Production

Authors: Islam Nowisser, Noha Farag, Mohamed El Azizi

Abstract:

Helicobacter pylori is a highly important human pathogen which inhabits about 50% of the population worldwide. Infection with this bacteria is very hard to treat, with high probability of recurrence. H. pylori causes severe gastric diseases, including peptic ulcer, gastritis, and gastric cancer, which has been linked to chronic inflammation. The infection has been reported to be associated with high levels of pro-inflammatory cytokines, especially IL-1β and TNF-α. The aim of the current study is to investigate the molecular mechanisms by which H. pylori activates NLRP3 inflammasome and its contribution to Il-1 β production in an innate cellular model. H. pylori PMSS1 and G27 standard strains, as well as the PMSS1 isogenic mutant strain PMSS1ΔVacA and G27ΔVacA, G27ΔCagA in addition to clinical isolates obtained from biopsy samples from the antrum and corpus mucosa of chronic gastritis patients, were used to establish infection in RAW-264.7 macrophages. The production levels of TNF-α and IL-1β was assessed using ELISA. Since expression of these cytokines is often regulated by the transcription factor complex, nuclear factor-kB (NF-kB), the activation of NF-κB in H. pylori infected cells was also evaluated by luciferase assay. Genomic DNA was extracted from bacterial cultures of H. pylori clinical isolates as well as the standard strains and their corresponding mutants, where they were evaluated for the cagA pathogenicity island and vacA expression. The correlation between these findings and expression of the cagA Pathogenicity Island and vacA in the bacteria was also investigated. The results showed IL-1β, and TNF-α production significantly increased in raw macrophages following H. pylori infection. The cagA+ and vacA+ H. pylori strains induced significant production of IL-1β compared to cagA- and vacA- strains. The activation pattern of NF-κB was correlated in the isolates to their cagA and vacA expression profiles. A similar finding could not be confirmed for TNF-α production. Our study shows the ability of H. pylori to activate NF-kB and induce significant IL-1β production as a possible mechanism for the augmented inflammatory response seen in subjects infected with cagA+ and vacA+ H. pylori strains that would lead to the progression to more severe form of the disease.

Keywords: Helicobacter pylori, IL-1β, inflammatory cytokines, nuclear factor KB, TNF-α

Procedia PDF Downloads 90
772 Comparison of Different Activators Impact on the Alkali-Activated Aluminium-Silicate Composites

Authors: Laura Dembovska, Ina Pundiene, Diana Bajare

Abstract:

Alkali-activated aluminium-silicate composites (AASC) can be used in the production of innovative materials with a wide range of properties and applications. AASC are associated with low CO₂ emissions; in the production process, it is possible to use industrial by-products and waste, thereby minimizing the use of a non-renewable natural resource. This study deals with the preparation of heat-resistant porous AASC based on chamotte for high-temperature applications up to 1200°C. Different fillers, aluminium scrap recycling waste as pores forming agent and alkali activation with 6M sodium hydroxide (NaOH) and potassium hydroxide (KOH) solution were used. Sodium hydroxide (NaOH) is widely used for the synthesis of AASC compared to potassium hydroxide (KOH), but comparison of using different activator for geopolymer synthesis is not well established. Changes in chemical composition of AASC during heating were identified and quantitatively analyzed by using DTA, dimension changes during the heating process were determined by using HTOM, pore microstructure was examined by SEM, and mineralogical composition of AASC was determined by XRD. Lightweight porous AASC activated with NaOH have been obtained with density in range from 600 to 880 kg/m³ and compressive strength from 0.8 to 2.7 MPa, but for AAM activated with KOH density was in range from 750 to 850 kg/m³ and compressive strength from 0.7 to 2.1 MPa.

Keywords: alkali activation, alkali activated materials, elevated temperature application, heat resistance

Procedia PDF Downloads 151
771 Effects of Active Muscle Contraction in a Car Occupant in Whiplash Injury

Authors: Nisha Nandlal Sharma, Julaluk Carmai, Saiprasit Koetniyom, Bernd Markert

Abstract:

Whiplash Injuries are usually associated with car accidents. The sudden forward or backward jerk to head causes neck strain, which is the result of damage to the muscle or tendons. Neck pain and headaches are the two most common symptoms of whiplash. Symptoms of whiplash are commonly reported in studies but the Injury mechanism is poorly understood. Neck muscles are the most important factor to study the neck Injury. This study focuses on the development of finite element (FE) model of human neck muscle to study the whiplash injury mechanism and effect of active muscle contraction on occupant kinematics. A detailed study of Injury mechanism will promote development and evaluation of new safety systems in cars, hence reducing the occurrence of severe injuries to the occupant. In present study, an active human finite element (FE) model with 3D neck muscle model is developed. Neck muscle was modeled with a combination of solid tetrahedral elements and 1D beam elements. Muscle active properties were represented by beam elements whereas, passive properties by solid tetrahedral elements. To generate muscular force according to inputted activation levels, Hill-type muscle model was applied to beam elements. To simulate non-linear passive properties of muscle, solid elements were modeled with rubber/foam material model. Material properties were assigned from published experimental tests. Some important muscles were then inserted into THUMS (Total Human Model for Safety) 50th percentile male pedestrian model. To reduce the simulation time required, THUMS lower body parts were not included. Posterior to muscle insertion, THUMS was given a boundary conditions similar to experimental tests. The model was exposed to 4g and 7g rear impacts as these load impacts are close to low speed impacts causing whiplash. The effect of muscle activation level on occupant kinematics during whiplash was analyzed.

Keywords: finite element model, muscle activation, neck muscle, whiplash injury prevention

Procedia PDF Downloads 320
770 Antiplatelet Activity of Nitrated Fatty Acids from Tomato Pomace

Authors: Lyanne Rodriguez, Eduardo Fuente, Andrés Trostchansky, Ivan Palomo

Abstract:

Cardiovascular diseases (CVD) are the leading cause of death in the world. The development of platelet-rich thrombi has been considered a trigger for acute cardiovascular events. A healthy diet, rich in fruit and vegetables, has been related to increased protection against cardiovascular events. Previous studies have observed that tomato pomace has a potent antiplatelet activity, due could be attributed to its high content of fatty acids (> 30%). It has been shown that unsaturated fatty acids can undergo endogenous intracellular nitration reactions during digestion after lipid consumption. Additionally, nitrated fatty acids (NO2-FA) can significantly reduce atherosclerotic lesion formation, inhibiting the expression of adhesion molecules on dysfunctional endothelium and platelet activation. In this work, we have proposed the nitration of fatty acids present in tomato pomace to improve its antiplatelet action. The gastric digestion of the tomato pomace allowed the nitration of the fatty acids, while by HPLC/MS/MS we were able to identify and quantify the nitrated fatty acids. The nitrated tomase extracts showed antiplatelet potential when platelets were stimulated with TRAP-6 and collagen. This activity was related to the presence of nitrated linoleic acid, which inhibited platelet activation by flow cytometry. The knowledge about the antiplatelet activity of nitrated fatty acids from tomato pomace will further develop new and more effective agents.

Keywords: cardiovascular, tomato extracts, nitrated fatty acids, antiplatelet activity

Procedia PDF Downloads 34
769 Kinetic Study of the Esterification of Unsaturated Fatty Acids from Salmon Oil (Salmosalar L.)

Authors: André Luis Lima de Oliveira, Vera Lúcia Viana do Nascimento, Victória Maura Silva Bermudez, Mauricio Nunes Kleinberg, João Carlos da Costa Assunção, José Osvaldo Beserra Carioca

Abstract:

The objective of this study was to synthesize a triglyceride with high content of unsaturated fatty acids from salmon oil (Salmo salar L.) by esterification with glycerol catalyzed dealuminized zeolite. A kinetic study was conducted to determine the reaction order and the activation energy. A statistical study was conducted to determine optimal reaction conditions. Initially, the crude oil was refined salmon physically and chemically. The crude oil was hydrolyzed and unsaturated free fatty acids were separated by urea complexation method. An experimental project to verify the parameters (temperature, glycerin and catalyst) with the greatest impact on the reaction was developed. In experiments aliquots were taken at predetermined times to measure the amount of free fatty acids. Pareto, surface, contour and hub graphs were used to determine the factors that maximized the reaction. According to the graphs the best reaction conditions were: temperature 80 ° C, the proportion glycerine/oil 5: 1 and 1% of catalyst. The kinetic data showed that the system was compatible with a second-order reaction. After analyzing the rate constant versus temperature charts a value of 85.31 kJ/mol was obtained for the reaction activation energy.

Keywords: esterification, kinect, oil, salmon

Procedia PDF Downloads 483
768 The Influence of Temperature on the Corrosion and Corrosion Inhibition of Steel in Hydrochloric Acid Solution: Thermodynamic Study

Authors: Fatimah Al-Hayazi, Ehteram. A. Noor, Aisha H. Moubaraki

Abstract:

The inhibitive effect of Securigera securidaca seed extract (SSE) on mild steel corrosion in 1 M HCl solution has been studied by weight loss and electrochemical techniques at four different temperatures. All techniques studied provided data that the studied extract does well at all temperatures, and its inhibitory action increases with increasing its concentration. SEM images indicate thin-film formation on mild steel when corroded in solutions containing 1 g L-1 of inhibitor either at low or high temperatures. The polarization studies showed that SSE acts as an anodic inhibitor. Both polarization and impedance techniques show an acceleration behaviour for SSE at concentrations ≤ 0.1 g L-1 at all temperatures. At concentrations ≥ 0.1 g L-1, the efficiency of SSE is dramatically increased with increasing concentration, and its value does not change appreciably with increasing temperature. It was found that all adsorption data obeyed Temkin adsorption isotherm. Kinetic activation and thermodynamic adsorption parameters are evaluated and discussed. The results revealed an endothermic corrosion process with an associative activation mechanism, while a comprehensive adsorption mechanism for SSE on mild steel surfaces is suggested, in which both physical and chemical adsorption are involved in the adsorption process. A good correlation between inhibitor constituents and their inhibitory action was obtained.

Keywords: corrosion, inhibition of steel, hydrochloric acid, thermodynamic study

Procedia PDF Downloads 59
767 A Prospective Study on Alkali Activated Bottom Ash-GGBS Blend in Paver Blocks

Authors: V. Revathi, J. Thaarrini, M. Venkob Rao

Abstract:

This paper presents a study on use of alkali activated bottom ash (BA) and ground granulated blast furnace slag (GGBS) blend in paver blocks. A preliminary effort on alkali-activated bottom ash, blast furnace slag based geopolymer (BA-GGBS-GP) mortar with river sand was carried out to identify the suitable mix for paver block. Several mixes were proposed based on the combination of BA-GGBS. The percentage ratio of BA:GGBS was selected as 100:0, 75:25, 50:50, 25:75 and 0:100 for the source material. Sodium based alkaline activators were used for activation. The molarity of NaOH was considered as 8M. The molar ratio of SiO2 to Na2O was varied from 1 to 4. Two curing mode such as ambient and steam curing 60°C for 24 hours were selected. The properties of paver block such as compressive strength split tensile strength, flexural strength and water absorption were evaluated as per IS15658:2006. Based on the preliminary study on BA-GGBS-GP mortar, the combinations of 25% BA with 75% GGBS mix for M30 and 75% BA with 25% GGBS mix for M35 grade were identified for paver block. Test results shows that the combination of BA-GGBS geopolymer paver blocks attained remarkable compressive strength under steam curing as well as in ambient mode at 3 days. It is noteworthy to know BA-GGBS-GP has promising future in the construction industry.

Keywords: bottom ash, GGBS, alkali activation, paver block

Procedia PDF Downloads 324
766 Molecular Simulation Study on the Catalytic Role of Silicon-Doped Graphene in Carbon Dioxide Hydrogenation

Authors: Wilmer Esteban Vallejo Narváez, Serguei Fomine

Abstract:

The theoretical investigation of Si-doped graphene nanoflakes (NFs) was conducted to understand their catalytic impact on CO₂ reduction using molecular hydrogen at the Density Functional Theory (DFT) level. The introduction of silicon by substituting carbon induces defects in the NF structure, resulting in a polyradical ground state. This silicon defect significantly boosts reactivity towards substrates, making Si-doped graphene NFs more catalytically active in CO₂ reduction to formic acid compared to silicene. Notably, Si-doped graphene demonstrates a preference for formic acid over carbon monoxide, mirroring the behavior of silicene. Furthermore, investigations into formic acid-to-formaldehyde and formaldehyde-to-methanol conversions reveal instances where Si-doped graphene outperforms silicene in terms of efficacy. In the final reduction step, the methanol-to-methane reaction unfolds in four stages, with the rate-determining step involving hydrogen transfer from silicon to methyl. Notably, the activation energy for this step is lower in Si-doped graphene compared to silicene. Consequently, Si-doped graphene NFs emerge as superior catalysts with lower activation energies overall. Remarkably, throughout these catalytic processes, Si-doped graphene maintains environmental stability, further highlighting its enhanced catalytic activity without compromising graphene's inherent stability.

Keywords: silicon-doped graphene, CO₂ reduction, DFT, catalysis

Procedia PDF Downloads 14
765 Preparation of Activated Carbon from Lignocellulosic Precursor for Dyes Adsorption

Authors: H. Mokaddem, D. Miroud, N. Azouaou, F. Si-Ahmed, Z. Sadaoui

Abstract:

The synthesis and characterization of activated carbon from local lignocellulosic precursor (Algerian alfa) was carried out for the removal of cationic dyes from aqueous solutions. The effect of the production variables such as impregnation chemical agents, impregnation ratio, activation temperature and activation time were investigated. Carbon obtained using the optimum conditions (CaCl2/ 1:1/ 500°C/2H) was characterized by various analytical techniques scanning electron microscopy (SEM), infrared spectroscopic analysis (FTIR) and zero-point-of-charge (pHpzc). Adsorption tests of methylene blue on the optimal activated carbon were conducted. The effects of contact time, amount of adsorbent, initial dye concentration and pH were studied. The adsorption equilibrium examined using Langmuir, Freundlich, Temkin and Redlich–Peterson models reveals that the Langmuir model is most appropriate to describe the adsorption process. The kinetics of MB sorption onto activated carbon follows the pseudo-second order rate expression. The examination of the thermodynamic analysis indicates that the adsorption process is spontaneous (ΔG ° < 0) and endothermic (ΔH ° > 0), the positive value of the standard entropy shows the affinity between the activated carbon and the dye. The present study showed that the produced optimal activated carbon prepared from Algerian alfa is an effective low-cost adsorbent and can be employed as alternative to commercial activated carbon for removal of MB dye from aqueous solution.

Keywords: activated carbon, adsorption, cationic dyes, Algerian alfa

Procedia PDF Downloads 193
764 Novel Nickel Complex Compound Reactivates the Apoptotic Network, Cell Cycle Arrest and Cytoskeletal Rearrangement in Human Colon and Breast Cancer Cells

Authors: Nima Samie, Batoul Sadat Haerian, Sekaran Muniandy, M. S. Kanthimathi

Abstract:

Colon and breast cancers are categorized as the most prevalent types of cancer worldwide. Recently, the broad clinical application of metal complex compounds has led to the discovery of potential therapeutic drugs. The aim of this study was to evaluate the cytotoxic action of a selected nickel complex compound (NCC) against human colon and breast cancer cells. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, MCF-7 and Hs 190.T cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content , measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results showed that our nickel complex compound displayed a potent suppressive effect on HT-29, WiDr, MCF-7 and Hs 190.T after 24 h of treatment with IC50 value of 2.02±0.54, 2.13±0.65, 3.76±015 and 3.14±0.45 µM respectively. This cytotoxic effect on normal cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the nickel complex compound. Activation of this pathway was further evidenced by significant activation of caspase 9 and 3/7.The nickel complex compound (NCC) was also shown activate the extrinsic pathways of apoptosis by activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. The results of this study suggest that the nickel complex compound is a potent anti-cancer agent inducing both intrinsic and extrinsic pathways as well as cell cycle arrest in colon and breast cancer cells.

Keywords: nickel complex, apoptosis, cytoskeletal rearrangement, colon cancer, breast cancer

Procedia PDF Downloads 282
763 Ethanol Extract of Potentilla pradoxa Nutt Inhibits LPS-induced Inflammatory Responses via NF-κB and AP-1 Inactivation

Authors: Hae-Jun Lee, Ji-Sun Shin, Kyung-Tae Lee

Abstract:

Potentilla species (Rosasease) have been used in traditional medicine to treat different ailment, disease or malady. In this study, we investigated the anti-inflammatory effects of ethanol extracts of NUTT (EPP) in lipopolysaccharide (LPS)-induced Raw 264.7 macrophages and septic mice. EPP suppressed LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production in LPS-induced Raw 264.7 macrophages. Consistent with these observations, EPP reduced the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) by downregulation of their promoter activities. EPP inhibited tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) at production and mRNA levels. Molecularly, EPP attenuated the LPS-induced transcriptional activity, and DNA-binding activity of nuclear factor-κB (NF-κB), and this was associated with a decrease of translocation and phosphorylation of p65 NF-κB by inhibiting the inhibitory κB-α (IκB-α) degradation and IκB kinase-α/β (IKK-α/β) phosphorylation. Furthermore, EPP suppressed the LPS-induced activation of activator protein-1 (AP-1) by reducing the expression of c-Fos and c-Jun in nuclear. EPP also reduced the phosphorylation of mitogen-activated protein kinase (MAPK), such as p38 MAPK and c-Jun N-terminal kinase/stress-activated protein kinase (JNK). In a sepsis model, pretreatment with EPP reduced the LPS-induced lethality. Collectively, these results suggest that the anti-inflammatory effects of EPP were associated with the suppression of NF-κB and AP-1 activation, and support its possible therapeutic role for the treatment of sepsis.

Keywords: anti-inflammation, activator protein-1, nuclear factor κB, Potentilla paradoxa Nutt

Procedia PDF Downloads 285
762 Modeling of CREB Pathway Induced Gene Induction: From Stimulation to Repression

Authors: K. Julia Rose Mary, Victor Arokia Doss

Abstract:

Electrical and chemical stimulations up-regulate phosphorylaion of CREB, a transcriptional factor that induces its target gene production for memory consolidation and Late Long-Term Potentiation (L-LTP) in CA1 region of the hippocampus. L-LTP requires complex interactions among second-messenger signaling cascade molecules such as cAMP, CAMKII, CAMKIV, MAPK, RSK, PKA, all of which converge to phosphorylate CREB which along with CBP induces the transcription of target genes involved in memory consolidation. A differential equation based model for L-LTP representing stimulus-mediated activation of downstream mediators which confirms the steep, supralinear stimulus-response effects of activation and inhibition was used. The same was extended to accommodate the inhibitory effect of the Inducible cAMP Early Repressor (ICER). ICER is the natural inducible CREB antagonist represses CRE-Mediated gene transcription involved in long-term plasticity for learning and memory. After verifying the sensitivity and robustness of the model, we had simulated it with various empirical levels of repressor concentration to analyse their effect on the gene induction. The model appears to predict the regulatory dynamics of repression on the L-LTP and agrees with the experimental values. The flux data obtained in the simulations demonstrate various aspects of equilibrium between the gene induction and repression.

Keywords: CREB, L-LTP, mathematical modeling, simulation

Procedia PDF Downloads 250
761 Inhibition of Glutamate Carboxypeptidase Activity Protects Retinal Ganglionic Cell Death Induced by Ischemia-Reperfusion by Reducing the Astroglial Activation in Rat

Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

Abstract:

Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

Procedia PDF Downloads 314
760 Cell-free Bioconversion of n-Octane to n-Octanol via a Heterogeneous and Bio-Catalytic Approach

Authors: Shanna Swart, Caryn Fenner, Athanasios Kotsiopoulos, Susan Harrison

Abstract:

Linear alkanes are produced as by-products from the increasing use of gas-to-liquid fuel technologies for synthetic fuel production and offer great potential for value addition. Their current use as low-value fuels and solvents do not maximize this potential. Therefore, attention has been drawn towards direct activation of these aliphatic alkanes to more useful products such as alcohols, aldehydes, carboxylic acids and derivatives. Cytochrome P450 monooxygenases (P450s) can be used for activation of these aliphatic alkanes using whole-cells or cell-free systems. Some limitations of whole-cell systems include reduced mass transfer, stability and possible side reactions. Since the P450 systems are little studied as cell-free systems, they form the focus of this study. Challenges of a cell-free system include co-factor regeneration, substrate availability and enzyme stability. Enzyme immobilization offers a positive outlook on this dilemma, as it may enhance stability of the enzyme. In the present study, 2 different P450s (CYP153A6 and CYP102A1) as well as the relevant accessory enzymes required for electron transfer (ferredoxin and ferredoxin reductase) and co-factor regeneration (glucose dehydrogenase) have been expressed in E. coli and purified by metal affinity chromatography. Glucose dehydrogenase (GDH), was used as a model enzyme to assess the potential of various enzyme immobilization strategies including; surface attachment on MagReSyn® microspheres with various functionalities and on electrospun nanofibers, using self-assembly based methods forming Cross Linked Enzymes (CLE), Cross Linked Enzyme Aggregates (CLEAs) and spherezymes as well as in a sol gel. The nanofibers were synthesized by electrospinning, which required the building of an electrospinning machine. The nanofiber morphology has been analyzed by SEM and binding will be further verified by FT-IR. Covalent attachment based methods showed limitations where only ferredoxin reductase and GDH retained activity after immobilization which were largely attributed to insufficient electron transfer and inactivation caused by the crosslinkers (60% and 90% relative activity loss for the free enzyme when using 0.5% glutaraldehyde and glutaraldehyde/ethylenediamine (1:1 v/v), respectively). So far, initial experiments with GDH have shown the most potential when immobilized via their His-tag onto the surface of MagReSyn® microspheres functionalized with Ni-NTA. It was found that Crude GDH could be simultaneously purified and immobilized with sufficient activity retention. Immobilized pure and crude GDH could be recycled 9 and 10 times, respectively, with approximately 10% activity remaining. The immobilized GDH was also more stable than the free enzyme after storage for 14 days at 4˚C. This immobilization strategy will also be applied to the P450s and optimized with regards to enzyme loading and immobilization time, as well as characterized and compared with the free enzymes. It is anticipated that the proposed immobilization set-up will offer enhanced enzyme stability (as well as reusability and easy recovery), minimal mass transfer limitation, with continuous co-factor regeneration and minimal enzyme leaching. All of which provide a positive outlook on this robust multi-enzyme system for efficient activation of linear alkanes as well as the potential for immobilization of various multiple enzymes, including multimeric enzymes for different bio-catalytic applications beyond alkane activation.

Keywords: alkane activation, cytochrome P450 monooxygenase, enzyme catalysis, enzyme immobilization

Procedia PDF Downloads 189
759 Resveratrol Ameliorates Benzo(a)Pyrene Induced Testicular Dysfunction and Apoptosis: Involvement of p38 MAPK/ATF2/iNOS Signaling

Authors: Kuladip Jana, Bhaswati Banerjee, Parimal C. Sen

Abstract:

Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems along with carcinogenesis in skin, prostate, ovary, lung and mammary glands. Our study was focused on elucidating the molecular mechanism of B(a)P induced male reproductive toxicity and its prevention with phytochemical like resveratrol. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of Wistar rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased Reactive Oxygen Species (ROS), altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38 mitogen activated protein kinase (p38MAPK), cytosolic translocation of cytochrome-c, upregulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of poly ADP ribose polymerase (PARP) and down regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like steroidogenic acute regulatory protein (StAR), cytochrome P450 IIA1 (CYPIIA1), 3β hydroxy steroid dehydrogenase (3β HSD), 17β hydroxy steroid dehydrogenase (17β HSD) showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis. Next we investigated the role of resveratrol on B(a)P induced male reproductive toxicity. Our study highlighted that resveratrol co-treatment with B(a)P maintained testicular redox potential, increased serum testosterone level and prevented steroidogenic dysfunction with enhanced expression of major testicular steroidogenic proteins (CYPIIA1, StAR, 3β HSD,17β HSD) relative to treatment with B(a)P only. Resveratrol suppressed B(a)P-induced testicular activation of p38 MAPK, ATF2, iNOS and ROS production; cytosolic translocation of Cytochome c and Caspase 3 activation thereby prevented oxidative stress of testis and inhibited apoptosis. Resveratrol co-treatment also decreased B(a)P-induced AhR protein level, its nuclear translocation and subsequent CYP1A1 promoter activation, thereby decreased protein and mRNA levels of testicular cytochrome P4501A1 (CYP1A1) and prevented BPDE-DNA adduct formation. Our findings cumulatively suggest that resveratrol prevents activation of B(a)P by modulating the transcriptional regulation of CYP1A1 and acting as an antioxidant thus prevents B(a)P-induced oxidative stress and testicular apoptosis.

Keywords: benzo(a)pyrene, resveratrol, testis, apoptosis, cytochrome P450 1A1 (CYP1A1), aryl hydrocarbon receptor (AhR), p38 MAPK/ATF2/iNOS

Procedia PDF Downloads 191