Search results for: enzyme treated

Authors: Devadrita Dey Sarkar

Abstract:

Oxidosqualene cyclase is a membrane bound enzyme in which helps in the formation of steroid scaffold in higher organisms. In a highly selective cyclization reaction oxidosqualene cyclase forms LANOSTEROL with seven chiral centres starting from the linear substrate 2,3-oxidosqualene. In humans OSC in cholesterol biosynthesis it represents a target for the discovery of novel anticholesteraemic drugs that could complement the widely used statins. The enzyme oxidosqualene: lanosterol cyclase (OSC) represents a novel target for the treatment of hypercholesterolemia. OSC catalyzes the cyclization of the linear 2,3-monoepoxysqualene to lanosterol, the initial four-ringed sterol intermediate in the cholesterol biosynthetic pathway. OSC also catalyzes the formation of 24(S), 25-epoxycholesterol, a ligand activator of the liver X receptor. Inhibition of OSC reduces cholesterol biosynthesis and selectively enhances 24(S),25-epoxycholesterol synthesis. Through this dual mechanism, OSC inhibition decreases plasma levels of low-density lipoprotein (LDL)-cholesterol and prevents cholesterol deposition within macrophages. The recent crystallization of OSC identifies the mechanism of action for this complex enzyme, setting the stage for the design of OSC inhibitors with improved pharmacological properties for cholesterol lowering and treatment of atherosclerosis. While studying and designing the inhibitor of oxidosqulene cyclase, I worked on the pdb id of 1w6k which was the most worked on pdb id and I used several methods, techniques and softwares to identify and validate the top most molecules which could be acting as an inhibitor for oxidosqualene cyclase. Thus, by partial blockage of this enzyme, both an inhibition of lanosterol and subsequently cholesterol formation as well as a concomitant effect on HMG-CoA reductase can be achieved. Both effects complement each other and lead to an effective control of cholesterol biosynthesis. It is therefore concluded that 2,3-oxidosqualene cyclase plays a crucial role in the regulation of intracellular cholesterol homeostasis. 2,3-Oxidosqualene cyclase inhibitors offer an attractive approach for novel lipid-lowering agents.

Keywords: anticholesteraemic, crystallization, statins, homeostasis

Procedia PDF Downloads 319

Authors: B. D. Nkosi, T. F. Mutavhatsindi, J. J. Baloyi, R. Meeske, T. M. Langa, I. M. M. Malebana, M. D. Motiang

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Potato hash (PH), a by-product from food production industry, contains 188.4 g dry matter (DM)/kg and 3.4 g water soluble carbohydrate (WSC)/kg DM, and was mixed with wheat bran (70:30 as is basis) to provide 352 g DM/kg and 315 g WSC/kg DM. The materials were ensiled with or without silage additives in 1.5L anaerobic jars (3 jars/treatment) that were kept at 25-280 C for 3 months. Four types of silages were produced which were: control (no additive, denoted as T1), celluclast enzyme (denoted as T2), emsilage bacterial inoculant (denoted as T3) and silosolve bacterial inoculant (denoted as T4). Three jars per treatment were opened after 3 months of ensiling for the determination of nutritive values, fermentation characteristics and aerobic stability. Aerobic stability was done by exposing silage samples to air for 5 days. The addition of enzyme (T2) was reduced (P<0.05) silage pH, fiber fractions (NDF and ADF) while increasing (P < 0.05) residual WSC and lactic acid (LA) production, compared to other treatments. Silage produced had pH of < 4.0, indications of well-preserved silage. Bacterial inoculation (T3 and T4) improved (P < 0.05) aerobic stability of the silage, as indicated by increased number of hours and lower CO2 production, compared to other treatments. However, the aerobic stability of silage was worsen (P < 0.05) with the addition of an enzyme (T2). Further work to elucidate these effects on nutrient digestion and growth performance on ruminants fed the silage is needed.

Keywords: by-products, digestibility, feeds, inoculation, ruminants, silage

Procedia PDF Downloads 411

Authors: Yasser Gaber, Mohamed Ismail, Serena Bisagni, Mohamad Takwa, Rajni Hatti-Kaul

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Enzymes are safe and selective catalysts. They skillfully catalyze chemical reactions; however, the native form is not usually suitable for industrial applications. Enzymes are therefore engineered by several techniques to meet the required catalytic task. Clopidogrel is recorded among the five best selling pharmaceutical in 2010 under the brand name Plavix. The commonly used route for production of the drug on an industrial scale is the synthesis of the racemic mixture followed by diastereomeric resolution to obtain the pure S isomer. The process consumes a lot of solvents and chemicals. We have evaluated a biocatalytic cleaner approach for asymmetric hydrolysis of racemic clopidogrel. Initial screening of a selected number of hydrolases showed only one enzyme EST to exhibit activity and selectivity towards the desired stereoisomer. As the crude EST is a mixture of several isoenzymes, a homology model of EST-1 was used in molecular dynamic simulations to study the interaction of the enzyme with R and S isomers of clopidogrel. Analysis of the geometric hindrances of the tetrahedral intermediates revealed a potential site for mutagenesis in order to improve the activity and the selectivity. Single point mutation showed dramatic increase in activity and inversion of the enantioselectivity (400 fold change in E value).

Keywords: biocatalysis, biotechnology, enzyme, protein engineering, molecular modeling

Procedia PDF Downloads 414

Authors: A. K. M. Kafi, S. N. Nina, Mashitah M. Yusoff

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In this work, we have described a new 3-dimensional (3D) network of cross-linked Horseradish Peroxidase/Carbon Nanotube (HRP/CNT) on a thiol-modified Au surface in order to build up the effective electrical wiring of the enzyme units with the electrode. This was achieved by the electropolymerization of aniline-functionalized carbon nanotubes (CNTs) and 4-aminothiophenol -modified-HRP on a 4-aminothiophenol monolayer-modified Au electrode. The synthesized 3D HRP/CNT networks were characterized with cyclic voltammetry and amperometry, resulting the establishment direct electron transfer between the redox active unit of HRP and the Au surface. Electrochemical measurements reveal that the immobilized HRP exhibits high biological activity and stability and a quasi-reversible redox peak of the redox center of HRP was observed at about −0.355 and −0.275 V vs. Ag/AgCl. The electron transfer rate constant, KS and electron transfer co-efficient were found to be 0.57 s-1 and 0.42, respectively. Based on the electrocatalytic process by direct electrochemistry of HRP, a biosensor for detecting H2O2 was developed. The developed biosensor exhibits excellent electrocatalytic activity for the reduction of H2O2. The proposed biosensor modified with HRP/CNT 3D network displays a broader linear range and a lower detection limit for H2O2 determination. The linear range is from 1.0×10−7 to 1.2×10−4M with a detection limit of 2.2.0×10−8M at 3σ. Moreover, this biosensor exhibits very high sensitivity, good reproducibility and long-time stability. In summary, ease of fabrication, a low cost, fast response and high sensitivity are the main advantages of the new biosensor proposed in this study. These obvious advantages would really help for the real analytical applicability of the proposed biosensor.

Keywords: redox enzyme, nanomaterials, biosensors, electrical communication

Procedia PDF Downloads 424

Authors: A. K. M. Kafi, S. N. Nina, Mashitah M. Yusoff

Abstract:

In this work, we have described a new 3-dimensional (3D) network of crosslinked Horseradish Peroxidase/Carbon Nanotube (HRP/CNT) on a thiol-modified Au surface in order to build up the effective electrical wiring of the enzyme units with the electrode. This was achieved by the electropolymerization of aniline-functionalized carbon nanotubes (CNTs) and 4-aminothiophenol -modified-HRP on a 4-aminothiophenol monolayer-modified Au electrode. The synthesized 3D HRP/CNT networks were characterized with cyclic voltammetry and amperometry, resulting the establishment direct electron transfer between the redox active unit of HRP and the Au surface. Electrochemical measurements reveal that the immobilized HRP exhibits high biological activity and stability and a quasi-reversible redox peak of the redox center of HRP was observed at about −0.355 and −0.275 V vs. Ag/AgCl. The electron transfer rate constant, KS and electron transfer co-efficient were found to be 0.57 s-1 and 0.42, respectively. Based on the electrocatalytic process by direct electrochemistry of HRP, a biosensor for detecting H2O2 was developed. The developed biosensor exhibits excellent electrocatalytic activity for the reduction of H2O2. The proposed biosensor modified with HRP/CNT 3D network displays a broader linear range and a lower detection limit for H2O2 determination. The linear range is from 1.0×10−7 to 1.2×10−4M with a detection limit of 2.2.0×10−8M at 3σ. Moreover, this biosensor exhibits very high sensitivity, good reproducibility and long-time stability. In summary, ease of fabrication, a low cost, fast response and high sensitivity are the main advantages of the new biosensor proposed in this study. These obvious advantages would really help for the real analytical applicability of the proposed biosensor.

Keywords: biosensor, nanomaterials, redox enzyme, thiol-modified Au surface

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Authors: Simbiat O. Ayilara-Akande, Soji Fakoya

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Quality and yield are always the target of farmers, including mushroom farmers. This study investigated how better Pleurotus pulmonarius can be obtained with the induction of mutagens into the process of spawn production in order to improve both the quality and the yield. Mushroom spawns were treated with ultraviolet radiation (UV) and hydroxylamine hydrochloride (HA) at different exposure times (2, 6, and 10 minutes) and different concentrations (10, 30, and 50Mm), respectively. The treated spawns were used to cultivate mushrooms on five substrates in the family of Gramineae viz: sorghum, rice, bamboo, sugarcane, and corn straws. Matured fruit bodies were harvested after a few weeks, and their parameters were taken and recorded. This study reveals a significant yield increase in mushroom grown on all the substrates when treated with ultraviolet radiation (UV) for 10 minutes and 6 minutes, respectively. Mushroom spawns treated with hydroxylamine hydrochloride showed a negative correlation in the yield with an increased in mutagen concentration. Hence, Ultraviolet light could be employed to enhance the quality and yield of mushroom production.

Keywords: mushroom, protein, mutagens, yield

Procedia PDF Downloads 113

Authors: Mahmoud M. Tash, S. Alkahtani

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The present study was undertaken to investigate the effect of pre-aging and aging parameters (time and temperature) on the mechanical properties of Al-Mg-Zn (7075) alloys. Ultimate tensile strength, 0.5% offset yield strength and % elongation measurements were carried out on specimens prepared from cast and heat treated 7075 alloys. Aging treatments were carried out for the as solution treated (SHT) specimens (after quenching in warm water). The specimens were aged at different conditions; Natural aging was carried out at room temperature for different periods of time. Double aging was performed for SHT conditions (pre-aged at different time and temperature followed by high temperature aging). Ultimate tensile strength, yield strength and % elongation as a function of different pre-aging and aging parameters are analysed to acquire an understanding of the effects of these variables and their interactions on the mechanical properties of Be-treated 7075 alloys.

Keywords: duplex aging treatment, mechanical properties, Al-Mg-Zn (7075) alloys, manufacturing

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Authors: Salina Y. Saddick

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Mild gestational hyperglycemia (MGH) is a very common complication of pregnancy that is characterized by intolerance to glucose. The association of angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism to MGH has been previously reported. In this study, we evaluated the association between ACE polymorphism and the risk of MGH in a Saudi population. We conducted a case-control study in a population of 100 MGH patients and 100 control subjects. ACE gene polymorphism was analyzed by the novel approach of tetraprimer amplification refractory mutation system (ARMS)-polymerase chain reaction (PCR). The frequency of ACE polymorphism was not associated with either alleles or genotypes in MGH patients. Glucose concentration was found to be significantly associated with the MGH group. Our study suggests that ACE genotypes were not associated with ACE polymorphism in a Saudi population.

Keywords: MGH, ACE, insertion polymorphism, deletion polymorphism

Procedia PDF Downloads 286

Authors: Muhammad Saiful Islam Khan, Yun Ji Kim

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Organophosphorus pesticides (OPPs) have been widely used to replace more persistent organochlorine pesticides because OPPs are more soluble in water and decompose rapidly in aquatic systems. Extensive uses of OPPs in modern agriculture are the major cause of the contamination of surface water. Regardless of the advantages gained by the application of pesticides in modern agriculture, they are a threat to the public health environment. With the aim of reducing possible health threats, several physical and chemical treatment processes have been studied to eliminate biological and chemical poisons from food stuff. In the present study, a micro-plasma device was used to reduce pesticides from the surface of food stuff. Pesticide free food items chosen in this study were perilla leaf, tomato, broccoli and blueberry. To evaluate the removal efficiency of pesticides, different washing methods were followed such as soaking with water, washing with bubbling water, washing with plasma-treated water and washing with chlorine water. 2 mL of 2000 ppm pesticide samples, namely, diazinone and chlorpyrifos were individuality inoculated on food surface and was air dried for 2 hours before treated with plasma. Plasma treated water was used in two different manners one is plasma treated water with bubbling the other one is aerosolized plasma treated water. The removal efficiency of pesticides from food surface was studied using HPLC. Washing with plasma treated water, aerosolized plasma treated water and chlorine water shows minimum 72% to maximum 87 % reduction for 4 min treatment irrespective to the types of food items and the types of pesticides sample, in case of soaking and bubbling the reduction is 8% to 48%. Washing with plasma treated water, aerosolized plasma treated water and chlorine water shows somewhat similar reduction ability which is significantly higher comparing to the soaking and bubbling washing system. The temperature effect of the washing systems was also evaluated; three different temperatures were set for the experiment, such as 22°C, 10°C and 4°C. Decreasing temperature from 22°C to 10°C shows a higher reduction in the case of washing with plasma and aerosolized plasma treated water, whereas an opposite trend was observed for the washing with chlorine water. Further temperature reduction from 10°C to 4°C does not show any significant reduction of pesticides, except for the washing with chlorine water. Chlorine water treatment shows lesser pesticide reduction with the decrease in temperature. The color changes of the treated sample were measured immediately and after one week to evaluate if there is any effect of washing with plasma treated water and with chlorine water. No significant color changes were observed for either of the washing systems, except for broccoli washing with chlorine water.

Keywords: chlorpyrifos, diazinone, pesticides, micro plasma

Procedia PDF Downloads 156

Authors: Promil Mehra, Nanthi Bolan, Jack Desbiolles, Risha Gupta

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Agricultural practices affect the soil physical and chemical properties, which in turn influence the soil microorganisms as a function of the soil biological environment. On the return to conventional tillage (CT) from continuing no-till (NT) cropping system, a very little information is available from the impact caused by the intermittent tillage on the soil biochemical properties from a short-term (2-year) study period. Therefore, the contribution made by different microorganisms (fungal, bacteria) was also investigated in order to find out the effective changes in the soil microbial activity under a South Australian dryland faring system. This study was conducted to understand the impact of microbial dynamics on the soil organic carbon (SOC) under NT and CT systems when treated with different levels of mulching (0, 2.5 and 5 t/ha). Our results demonstrated that from the incubation experiment the cumulative CO2 emitted from CT system was 34.5% higher than NT system. Relatively, the respiration from surface layer (0-10 cm) was significantly (P<0.05) higher by 8.5% and 15.8 from CT; 8% and 18.9% from NT system w.r.t 10-20 and 20-30 cm respectively. Further, the dehydrogenase enzyme activity (DHA) and microbial biomass carbon (MBC) were both significantly lower (P<0.05) under CT, i.e., 7.4%, 7.2%, 6.0% (DHA) and 19.7%, 15.7%, 4% (MBC) across the different mulching levels (0, 2.5, 5 t/ha) respectively. In general, it was found that from both the tillage system the enzyme activity and MBC decreased with the increase in depth (0-10, 10-20 and 20-30 cm) and with the increase in mulching rate (0, 2.5 and 5 t/ha). From the perspective of microbial stress, there was 28.6% higher stress under CT system compared to NT system. Whereas, the microbial activity of different microorganisms like fungal and bacterial activities were determined by substrate-induced inhibition respiration using antibiotics like cycloheximide (16 mg/gm of soil) and streptomycin sulphate (14 mg/gm of soil), by trapping the CO2 using an alkali (0.5 M NaOH) solution. The microbial activities were confirmed through platting technique, where it was that found bacterial activities were 46.2% and 38.9% higher than fungal activity under CT and NT system. In conclusion, it was expected that changes in the relative abundance and activity of different microorganisms (bacteria and fungi) under different tillage systems could significantly affect the C cycling and storage due to its unique structures and differential interactions with the soil physical properties.

Keywords: tillage, soil respiration, MBC, fungal-bacterial activity

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Authors: Iveta Nováková, Martin-Andrè S. Husby, Boy-Arne Buyle

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The building industry has one of the most significant contributions to global warming due to the production of building materials, transportation, building activities, and demolition of structures when they reach the end of their life. Implementation of circular material flow and circular economy can significantly reduce greenhouse gasses and simultaneously reduce the need for natural resources. The use of recycled concrete aggregates (RCA) is one of the possibilities for reducing the depletion of raw materials for concrete production. Concrete is the most used building material worldwide, and aggregates constitute 70% of its volume. RCA can replace a certain amount of natural aggregates (NA), and concrete will still perform as required. The aim of this scientific paper is to evaluate RCA properties with and without mechanical treatment. Analysis of RCA itself will be followed by compressive strength of concrete containing various amounts of treated and non-treated RCA. Results showed improvement in compressive strength of the mix with mechanically treated RCA compared to standard RCA, and even the strength of concrete with mechanically treated RCA in dose 50% of coarse aggregates was higher than the reference mix by 4%. Based on obtained results, it can be concluded that integration of RCA in industrial concrete production is feasible, at a replacement ratio of 50% for mechanically treated RCA and 30% if untreated RCA is used, without affecting the compressive strength negatively.

Keywords: recycled concrete aggregates, mechanical treatment, aggregate properties, compression strength

Procedia PDF Downloads 206

Authors: Nathalie Baeza-Kallee, Raphaël Bergès, Victoria Hein, Stéphanie Cabaret, Jeremy Garcia, Abigaëlle Gros, Emeline Tabouret, Aurélie Tchoghandjian, Carole Colin, Dominique Figarella-Branger

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Glioblastoma (GBM) contains cancer stem cells that are resistant to treatment. GBM cancer stem cell expresses glycolipids recognized by the A2B5 antibody. A2B5, induced by the enzyme ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialyl transferase 3 (ST8Sia3), plays a crucial role in the proliferation, migration, clonogenicity, and tumorigenesis of GBM cancer stem cells. Our aim was to characterize the resulting effects of neuraminidase that remove A2B5 in order to target GBM cancer stem cells. To this end, we set up a GBM organotypic slice model; quantified A2B5 expression by flow cytometry in U87-MG, U87-ST8Sia3, and GBM cancer stem cell lines, treated or not by neuraminidase; performed RNAseq and DNA methylation profiling; and analyzed the ganglioside expression by liquid chromatography-mass spectrometry in these cell lines, treated or not with neuraminidase. Results demonstrated that neuraminidase decreased A2B5 expression, tumor size, and regrowth after surgical removal in the organotypic slice model but did not induce a distinct transcriptomic or epigenetic signature in GBM CSC lines. RNAseq analysis revealed that OLIG2, CHI3L1, TIMP3, TNFAIP2, and TNFAIP6 transcripts were significantly overexpressed in U87-ST8Sia3 compared to U87-MG. RT-qPCR confirmed these results and demonstrated that neuraminidase decreased gene expression in GBM cancer stem cell lines. Moreover, neuraminidase drastically reduced ganglioside expression in GBM cancer stem cell lines. Neuraminidase, by its pleiotropic action, is an attractive local treatment against GBM.

Keywords: cancer stem cell, ganglioside, glioblastoma, targeted treatment

Procedia PDF Downloads 47

Authors: M. Tash, S. Alkahtani

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The present study was undertaken to investigate the effect of pre-aging and aging parameters (time and temperature) on the mechanical properties of Al-Mg-Zn (7075) alloys. Ultimate tensile strength, 0.5% offset yield strength and % elongation measurements were carried out on specimens prepared from cast and heat treated 7075 alloys. Duplex aging treatments were carried out for the as solution treated (SHT) specimens (pre-aged at different time and temperature followed by high temperature aging). A statistical design of experiments (DOE) approach using fractional factorial design was applied to determine the influence of controlling variables of pre-aging and aging treatment parameters and any interactions between them on the mechanical properties of 7075 alloys. A mathematical models are developed to relate the alloy ultimate tensile strength, yield strength and % elongation with the different pre-aging and aging parameters i.e. Pre-aging Temperature (PA T0C), Pre-aging time (PA t h), Aging temperature (AT0C), Aging time (At h), to acquire an understanding of the effects of these variables and their interactions on the mechanical properties of be-treated 7075 alloys.

Keywords: aging heat Treatment, tensile properties, be-treated cast Al-Mg-Zn (7075) alloys, experimental correlation

Procedia PDF Downloads 247

Authors: L. L. Tundisi, A. Pessoa Jr., E. B. Tambourgi, E. Silveira, P. G. Mazzola

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L-asparaginase (L-ASNase) is the gold standard treatment for acute lymphoblastic leukemia that mainly affects pediatric patients; treatment increases survival from 20% to 90%. The characterization of other L-Asparaginases, apart from the most used from Escherichia coli and Erwinia chrysanthemi, has been reported, but the choice of the most appropriate is still under debate. This choice should be based on its pharmacokinetics, immune hypersensitivity, doses, prices, pharmacodynamics. The main factors influencing the antileukemic activity of ASNase are enzymatic activity, Km, glutaminase activity, clearance of the enzyme and development of resistance. However, most of the commercialized enzyme present an intrinsic glutaminase activity, which is responsible for some side effects. In this study, glutaminase free asparaginase produced from Aspergillus oryzae was precipitated in different percentages of ethanol (0–80%), until optimum ethanol concentration of 60% (w/w) was found. Following, precipitation of crude L-ASNase was performed in a single step, using 60% (w/w) ethanol, under constant agitation and temperature. It presented activity of 135.45 U/mg and after gel filtration chromatography with Sephadex G-the enzymatic activity was 322.02 U/mg. The apparent molecular mass of the purified L-ASNase fraction was estimated by 10% SDS-PAGE. Proteins were stained with Coomassie Brilliant Blue R-250. The molar mass range was from 10 kDa to 250 kDa. L-ASNase from Aspergillus oryzae was characterized aiming possible therapeutic use. Four different buffers (phosphate-citrate buffer pH 2.6 to 5.8; phosphate buffer pH 5.8 to 7.4; Tris - HCl pH 7.4 to 9.0; and carbonate buffer pH 9.8 to 10.6) were used to measure the optimum pH for L-ASNase activity. The optimum temperature for enzyme activity was measured at optimal pH conditions (Tris-HCl and phosphate buffer, pH 7.4) at different temperatures ranging from 5 to 55°C. All activities were calculated by quantifying the free ammonia, using the Nessler reagent. The kinetic parameters calculation, e.g. Michaelis-Menten constant (Km), maximum velocity (Vmax) and Hills coefficient (n), were performed by incubating the enzyme in different concentrations of the substrate at optimum conditions of pH and fitted on Hill’s equation. This glutaminase free asparaginase showed a low Km (3.39 mM and 3.81 mM) and enzymatic activity of 135.45 U/mg after precipitation with ethanol. After gel filtration chromatography it rose to 322.02 U/mg. Optimum activity was found between pH 5.8 - 9.0, best activity results with phosphate buffer pH 7.4 and Tris-HCl pH 7.4 and showed activity from 5°C to 55°C. These results indicate that L-ASNase from A. oryzae has the potential for human use.

Keywords: biopharmaceuticals, bioprocessing, bioproducts, biotechnology, enzyme activity, ethanol precipitation

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Authors: Sanjiv Kumar Soni, Chetna Janveja

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The approach of utilizing zero cost kitchen waste residues for growing suitable strains of fungi for the induction of a cocktail of hydrolytic enzymes and ethanol generation has been validated in the present study with the objective of developing an indigenous biorefinery for low cost bioethanol production with the generation of zero waste. Solid state fermentation has been carried out to evaluate the potential of various steam pretreated kitchen waste residues as substrates for the co-production of multiple carbohydrases including cellulases, hemicellulases, pectinase and amylases by a locally isolated strain of Aspergillus niger C-5. Of all the residues, potato peels induced the maximum yields of all the enzyme components corresponding to 64.0±1.92 IU of CMCase, 17.0±0.54 IU of FPase , 42.8±1.28 IU of β-glucosidase, 990.0±28.90 IU of xylanase, 53.2±2.12 IU of mannanase, 126.0±3.72 IU of pectinase, 31500.0±375.78 IU of α-amylase and 488.8±9.82 IU of glucoamylase/g dry substrate respectively. Saccharification of various kitchen refuse residues using inhouse produced crude enzyme cocktail resulted in the release of 610±10.56, 570±8.89, 435±6.54, 475±4.56, 445±4.27, 385±4.49, 370±6.89, 490±10.45 mg of total reducing sugars/g of dried potato peels, orange peels, pineapple peels, mausami peels, onion peels, banana stalks, pea pods and composite mixture respectively revealing carbohydrate conversion efficiencies in the range of 97.0-99.4%. After fermentation of released hexoses by Saccharomyces cerevisae, ethanol yields ranging from 80-262 mL/ kg of dry residues were obtained. The study has successfully evaluated the valorization of kitchen garbage, a highly biodegradable component in Municipal Solid Waste by using it as a substrate for the in-house co-production of multiple carbohydrases and employing the steam treated residues as a feed stock for bioethanol production. Such valorization of kitchen garbage may reduce the level of Municipal Solid Waste going into land-fills thus lowering the emissions of greenhouse gases. Moreover, the solid residue left after the bioconversion may be used as a biofertilizer for improving the fertility of the soils.

Keywords: kitchen waste, bioethanol, solid waste, bioconversion, waste management

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Authors: Haneef Ur Rehman, Afsheen Aman, Abdul Hameed Baloch, Shah Ali Ul Qader

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Pectinase is a heterogeneous group of enzymes that catalyze the hydrolysis of pectin substances and widely has been used in food and textile industries. In current study, pectinase from B. licheniformis KIBGE-IB21 was immobilized within different polymers (calcium alginate beads, polyacrylamide gel and agar-agar matrix) to enhance its catalytic properties. Polyacrylamide gel was found to be most promising one and gave maximum (89%) immobilization yield. While less immobilization yield was observed in case of calcium alginate beads that only retained 46 % activity. The reaction time for maximum pectinolytic activity was increased from 5.0 to 10 minutes after immobilization. The temperature of pectinase for maximum enzyme activity was increased from 45 °C to 50 °C and 55 °C when it was immobilized within agar-agar and calcium alginate beads, respectively. The optimum pH of pectinase didn’t alter when it was immobilized within polyacrylamide gel and calcium alginate beads, but in case of agar-agar it was changed from pH 10 to pH 9.0. Thermal stability of pectinase was improved after immobilization and immobilized pectinase showed higher toleration against different temperatures as compared to free enzyme. It can be concluded that the entrapment is a simple, single step and promising procedure to immobilized pectinase within different synthetic and non-synthetic polymers and enhanced its catalytic properties.

Keywords: pectinase, characterization immobilization, polyacrylamide, agar-agar, calcium alginate beads

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Authors: Fatima Zohra Ibn Majdoub Hassani, Ivan Lavandera, Joseph Kreit

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This study explored the effects of different organic solvents, temperature, and the amount of glycerol on the alcohol dehydrogenase (ADH)-catalysed stereoselective reduction of different ketones. These conversions were then analyzed by gas chromatography. It was found that when the amount of deep eutectic solvents (DES) increases, it can improve the stereoselectivity of the enzyme although reducing its ability to convert the substrate into the corresponding alcohol. Moreover, glycerol was found to have a strong stabilizing effect on the ADH from Ralstonia sp. (E. coli/ RasADH). In the case of organic solvents, it was observed that the best conversions into the alcohols were achieved with DMSO and hexane. It was also observed that temperature decreased the ability of the enzyme to convert the substrates into the products and also affected the selectivity. In addition to that, the recycling of DES up to three times gave good conversions and enantiomeric excess results and glycerol showed a positive effect in the stability of various ADHs. Using RasADH, a good conversion and enantiomeric excess into the S-alcohol were obtained. It was found that an enhancement of the temperature disabled the stabilizing effect of glycerol and decreased the stereoselectivity of the enzyme. However, for other ADHs a temperature increase had an opposite positive effect, especially with ADH-T from Thermoanaerobium sp. One of the objectives of this study was to see the effect of cofactors such as NAD(P) on the biocatlysis activities of ADHs.

Keywords: alcohol dehydrogenases, DES, gas chromatography, RasADH

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Authors: Salem Abdalla

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Objective: Dipyrone is a widely used, well tolerated analgesic drug which, however, is compromised by agranulocytosis as an adverse effect. Subsequent to no enzymatic hydrolysis, the primary metabolic step is N-demethylation of 4-methylaminoantipyrine (4-MAA) to 4-aminoantipyrine (4-AA). The aim of the present study was to identify the cytochrome P-450 enzyme (CYP) mediating this reaction. Methods: We identified the relevant CYP using virus expressed isolated rat liver microsomes with chemical inhibition studies. The substrate of 4-methylaminantipyrine was employed at six different concentrations (25, 50, 100, 400, 800, and 1200 µmol/l) with varying concentrations of selective inhibitors of CYP1A2 (furafylline, fluvoxamine), CYP3A4 (ketoconazole), CYP2A6 (coumarin), CYP2D6 (quinidine), CYP2C19 (omeprazole, fluvoxamine, tranylcypromine), CYP2C9 (sulfaphenazole), and CYP1A1 (alpha-naphthoflavone). 4-MAA and 4-AA were analyzed by HPLC, and enzyme kinetic parameters (Km and Vmax) were determined by regression (Sigma plot 9.0). Results: The N-demethylation of 4-MAA by microsomes prepared from baculovirus-expressing human CYP was pronounced with CYP2C19. Intrinsic clearances of the most active enzymes were 0.092, 0.027, and 0.026 for the CYP enzymes 2C19, 2D6, and 1A2, respectively. Metabolism by rat liver microsomes was strongly inhibited by omeprazole (IC50 of 0.05). Conclusion: The enzyme CYP2C19 apparently has an important role in N-demethylation of 4-methylaminoantipyrine which should be further analyzed in clinical studies and which may also be interesting concerning the agranulocytosis.

Keywords: dipyrone, 4-methylaminoantipyrine (4-MAA), 4- aminoantipyrine (4-AA), metabolism, human CYP2C19

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Authors: A. Goma Amira, U. E. Mahrous

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Measuring the effect of perinatal lead exposure on learning ability of offspring is considered as a sensitive and selective index for providing an early marker for central nervous system damage produced by this toxic metal. A total of 35 Sprague-Dawley adult rats were used to investigate the effect of lead acetate toxicity on behavioral patterns of adult female rats and learning ability of offspring. Rats were allotted into 4 groups, group one received 1g/l lead acetate (n=10), group two received 1.5g/l lead acetate (n=10), group three received 2g/l lead acetate in drinking water (n=10), and control group did not receive lead acetate (n=5) from 8th day of pregnancy till weaning of pups. The obtained results revealed a dose-dependent increase in the feeding time, drinking frequency, licking frequency, scratching frequency, licking litters, nest building, and retrieving frequencies, while standing time increased significantly in rats treated with 1.5g/l lead acetate than other treated groups and control. On the contrary, lying time decreased gradually in a dose-dependent manner. Moreover, movement activities were higher in rats treated with 1g/l lead acetate than other treated groups and control. Furthermore, time spent in closed arms was significantly lower in rats given 2g/l lead acetate than other treated groups, while they spent significantly much time spent in open arms than other treated groups which could be attributed to occurrence of adaptation. Furthermore, number of entries in open arms was-dose dependent. However, the ratio between open/closed arms revealed a significant decrease in rats treated with 2g/l lead acetate than the control group.

Keywords: lead toxicity, rats, learning ability, behavior

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Authors: Wegene Demisie Jima

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Leather is a durable and flexible material used for various purposes including clothing, footwear, upholstery and gloves. However, the use of leather for smart product applications is a challenge since it is electrically insulating material. Here, we report a simple method to produce conducting glove leathers using an in-situ polymerization of pyrrole. The concentrations of pyrrole, ferric chloride and anthraquinone-2-sulfonic acid sodium salt monohydrate were optimized to produce maximum conductivity in the treated leathers. The coating of polypyrrole in the treated leathers was probed using FT-IR, X-ray diffraction and electron microscopic analysis. FTIR confirms that the formation of polypyrrole on the leather surface as well as presence of prominent N-C stretching band. X-ray diffraction analysis suggests para-crystallinity in the PPy-treated leathers.We further demonstrate that the treated leathers, with maximum conductivity of 7.4 S/cm, can be used for making conductive gloves for operating touch-screen devices apart from other smart product applications.

Keywords: electrical conductivity, in-situ polymerization, pyrrole, smart product

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Authors: Hina Gul Rajpoot, Wazeer Hussain Solangi

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Experimental Design (DOE) economically maximizes information; we deliberately change one or more process variables (looms) in order to observe the effect the changes have on one or more response fabric properties. In DOE obtained data can be analyzed to yield valid and objective conclusions. An Experimental Design is lying out of a detailed experimental plan in advance and maximizes the amount of "information" that can be obtained for a given amount of experimental. Fabric of 36 inches having following weaves was used. 3/1 twill, warp cotton (10.5 den), weft Lycra (16 spandex * 70 den) Ends per inch86, Picks per inch 52 and washes process includes Stone wash, Rinse wash, Bleaching and Enzyme wash. Once the samples were ready, they were subjected to tensile and tear strength tests, for these two kinds of samples were considered. One washed fabric samples of warp direction type and other type of the samples was weft direction. Then five samples from each were considered for tensile and teat strength tests separately then takes the mean value. The results found that the lowest strength damaged in the weft direction observed by tensile strength test & Enzyme wash. Maximum breaking load of the enzyme washed fabric sample was 42 kg.

Keywords: twill, indigo dye, tear strength, loom, ball warp, denier or den, seam, waist band, pilling, selvage

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Authors: Supawadee Phetkhajone, Wisuwat Songnuan

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Metalaxyl is one of the most common and effective fungicides used to control Phytophthora palmivora infection in durian (Durio zibethinus L.). The efficacy of metalaxyl residue in durian under greenhouse condition was evaluated using bioassay. Durian seedlings were treated with 2 methods of application, spraying, and soil drenching of metalaxyl, at recommended concentration (1000 mg/L). Mock treated samples were treated with 0.1% Tween20 and water for spraying and soil drenching methods, respectively. The experiment was performed in triplicates. Leaves were detached from treated plants at 0, 1, 7, 15, 20, 30, and 60 days after application, inoculated with metalaxyl-resistant and metalaxyl-sensitive isolates of P. palmivora, and incubated in a high humidity chamber for 5 days at room temperature. Metalaxyl efficacy was determined by measuring the lesion size on metalaxyl treated and mock treated samples. The results showed that metalaxyl can control metalaxyl-sensitive isolate of P. palmivora for at least 30 days after application in both methods of application. The metalaxyl-resistant isolate was not inhibited in all treatments. Leaf samples from spraying method showed larger lesions compared to soil drench method. These results demonstrated that metalaxyl applications, especially soil drenching methods showed high efficacy to control metalaxyl-sensitive isolates of P. palmivora, although it cannot control metalaxyl-resistant isolates of P. palmivora in all treatments. These qualitative data indicate that metalaxyl may suitable to control metalaxyl-sensitive isolates of P. palmivora infection.

Keywords: bioassay, degradation, durian, metalaxyl

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Authors: Ji Hye Im, Nguyen T. T. Phuong, Keon Wook Kang

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Estrogens are important for the development and growth of estrogen receptor (ER)-positive breast cancer, for which anti-estrogen therapy is one of the most effective treatments. However, its efficacy can be limited by either de novo or acquired resistance. Aromatase is a key enzyme for the biosynthesis of estrogens, and inhibition of this enzyme leads to profound hypoestrogenism. Here, we found that the basal expression and activity of aromatase were significantly increased in tamoxifen (TAM)-resistant human breast cancer (TAMR-MCF-7) cells compared to control MCF-7 cells. We further revealed that aromatase immunoreactivity in tumor tissues was increased in recurrence group after TAM therapy compared to non-recurrence group after TAM therapy. Phosphorylation of Akt, extracellular signal-regulated kinase (ERK), and p38 kinase were all increased in TAMR-MCF-7 cells. Inhibition of phosphoinositide 3-kinase (PI3K) suppressed the transactivation of the aromatase gene and its enzyme activity. Furthermore, we have also shown that PI3K/Akt-dependent cAMP-response element binding protein (CREB) activation was required for the enhanced expression of aromatase in TAMR-MCF-7 cells. Our findings suggest that aromatase expression is up-regulated in TAM-resistant breast cancer via PI3K/Akt-dependent CREB activation.

Keywords: TAMR-MCF-7, CREB, estrogen receptor, aromatase

Procedia PDF Downloads 385

Authors: Hamid Hatami

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This experiment was included two cultivars i.e. Habbit and L17 (Main factor) with six fertilizer treatments i.e. control, seed inoculated with rhyzobium, base nitrogen + top-dress urea at R2 stage, base nitrogen + seed inoculated with rhyzobium + top-dress nitrogen at R2 stage, seed treated with humax + top-dress humax at R2 stage, base nitrogen + seed treated with humax + top-dress humax at R2 stage (sub factors ), as split-plot on the basis of RCBD with 3 replications at 2014. Treatment fertilizer of base nitrogen + seed treated with humax + top- dress humax at R2 stage and base nitrogen + top-dress urea in R2 stage had a significant superiority than the other fertilizer treatment in biological yield. L17 and Habbit with base nitrogen + seed treated with humax + top-dress humax in R2 stage and yield economical 5600 and 5767 kg/ha respectively, showed the most economical yield and Habbit cultivar with control and economical yield 3085 kg/ha showed the least economical yield among all the treatments. Results showed that fertilizer treatment of base nitrogen + seed treated with humax + top-dress humax in R2 stage and Habbit variety were suitable in this study.

Keywords: soybean, humax, rhyzobium, habbit

Procedia PDF Downloads 420

Authors: Md. Z. Alam, Amal A. Elgharbawy, Muhammad Moniruzzaman, Nassereldeen A. Kabbashi, Parveen Jamal

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The enzymatic hydrolysis of lignocellulosic biomass is one of the obstacles in the process of sugar production, due to the presence of lignin that protects the cellulose molecules against cellulases. Although the pretreatment of lignocellulose in ionic liquid (IL) system has been receiving a lot of interest; however, it requires IL removal with an anti-solvent in order to proceed with the enzymatic hydrolysis. At this point, introducing a compatible cellulase enzyme seems more efficient in this process. A cellulase enzyme that was produced by Trichoderma reesei on palm kernel cake (PKC) exhibited a promising stability in several ILs. The enzyme called PKC-Cel was tested for its optimum pH and temperature as well as its molecular weight. One among evaluated ILs, 1,3-diethylimidazolium dimethyl phosphate [DEMIM] DMP was applied in this study. Evaluation of six factors was executed in Stat-Ease Design Expert V.9, definitive screening design, which are IL/ buffer ratio, temperature, hydrolysis retention time, biomass loading, cellulase loading and empty fruit bunches (EFB) particle size. According to the obtained data, IL-enzyme system shows the highest sugar concentration at 70 °C, 27 hours, 10% IL-buffer, 35% biomass loading, 60 Units/g cellulase and 200 μm particle size. As concluded from the obtained data, not only the PKC-Cel was stable in the presence of the IL, also it was actually stable at a higher temperature than its optimum one. The reducing sugar obtained was 53.468±4.58 g/L which was equivalent to 0.3055 g reducing sugar/g EFB. This approach opens an insight for more studies in order to understand the actual effect of ILs on cellulases and their interactions in the aqueous system. It could also benefit in an efficient production of bioethanol from lignocellulosic biomass.

Keywords: cellulase, hydrolysis, lignocellulose, pretreatment

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Authors: Frieda Eivazi, Nikita L. Mullings

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Soils are recipient for surfactants in herbicide formulations. Surfactants entering the soil environment can possibly disrupt different chemical, physical and biological interactions. Therefore, it is critical that we understand the fate, behavior and transport of surfactants upon entering the soil. A comprehensive study was conducted to examine effect of surfactants on nutrient uptake, microbial community, and enzyme activity. The research was conducted in the greenhouse growing corn (Zea mays) as a test plant in a factorial experiment (three surfactants at two different rates with control, and three herbicides) organized as randomized blocked design. Surfactants evaluated were Activator 90, Agri-Dex, and Thrust; herbicides were glyphosate, atrazine, and bentazon. Treatments examined were surfactant only, herbicide only, and surfactant + herbicide combinations. Corn was planted in fertilized soils (silt loam and silty clay) with moisture content maintained at the field capacity for optimum growth. This paper will report results of above mentioned treatments on acid phosphatase, beta-glucosidase, arylsulfatase, beta-glucosaminidase, and dehydrogenase activities. In general, there were variations in the enzyme activities with some inhibition and some being enhanced by the treatments. Activator 90 appeared to have the highest inhibitory effect on enzymatic activities. Atrazine application significantly decreased the activities of acid phosphatase, beta-glucosidase, and dehydrogenase in both soils; however, combination of Atrazine + Agridex increased the acid phosphatase activity while significantly inhibiting the other enzyme activities in soils. It was concluded that long-term field studies are needed to validate changes in nutrient uptake, microbial community and enzyme activities due to surfactant-herbicide combination effects.

Keywords: herbicides, nutrient cycling, soil enzymes, surfactant

Procedia PDF Downloads 219

Authors: Mallika Supa-Aksorn, Buaream Maneewan, Jiraporn Rojtinnakorn

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For animals, trypsin and chymotrypsin are the 2 proteases that play the important role in protein digestion and involving in growth rate. In many animals, these two enzymes are indicated as growth parameter by feed. Although enzyme assay at optimal condition is significant for its accuracy activity determination. There is less report of trypsin and chymotrypsin. Therefore, in this study, optimization of pH and temperature for trypsin (T) and chymotrypsin (C) in economic species; i.e. Nile tilapia (Oreochromis niloticus), sand goby (Oxyeleotoris marmoratus), giant freshwater prawn (Macrobachium rosenberchii) and native chicken (Gallus gallus) were investigated. Each enzyme of each species was assaying for its specific activity with variation of pH in range of 2-12 and temperature in range of 30-80 °C. It revealed that, for Nile tilapia, T had optimal condition at pH 9 and temperature 50-80 °C, whereas C had optimal condition at pH 8 and temperature 60 °C. For sand goby, T had optimal condition at pH 7 and temperature of 50 °C, while C had optimal condition at pH 11 and temperature of 70-75 °C. For juvenile freshwater prawn, T had optimal condition at pH 10-11 and temperature of 60-65 °C, C had optimal condition at pH 8 and temperature of 70°C. For starter native chicken, T has optimal condition at pH 7 and temperature of 70 °C, whereas C had o optimal condition at pH 8 and temperature of 60°C. This information of optimal conditions will be high valuable in further for, actual enzyme measurement of T and C activities that benefit for growth and feed analysis.

Keywords: trypsin, chymotrypsin, Oreochromis niloticus, Oxyeleotoris marmoratus, Macrobachium rosenberchii, Gallus gallus

Procedia PDF Downloads 232

Authors: Pardis Abolghasemi, Benyamin Hatamsaz

Abstract:

Background: Diabetic nephropathy is a serious health problem described by specific kidney structure and functional disturbance. Renoprotective effects of the stem cells secretase have been shown in many kidney diseases. The aim is to evaluate the capability of human Wharton’s jelly mesenchymal stem cells conditioned media (hWJMSCs-CM) to alleviate DN in streptozotocin (STZ)-induced diabetes. Methods: Diabetic nephropathy was induced by injection of STZ (60 mg/kg, IP) in twenty rats. Conditioned media was extracted from hWJMSCs at third passages. At week 8, diabetic rats were divided into two groups: treated (hWJMSCs-CM, 500 μl/rat for three weeks, IP) and not treated (DN). In the 11th week, three groups (control, DN and DN+hWJMSCs-CM) were kept in metabolic cages and urine was collected for 24h. Blood pressure (BP) and heart rate (HR) were continuously recorded. The serum samples were maintained for measuring BUN, Cr and angiotensin-converting enzyme (ACE) activity. The left kidney was kept at -80°C for ACE activity assessment. The right kidney and pancreas were used for histopathologic evaluation. Result: Diabetic nephropathy was detected by microalbuminuria and increased albumin/creatinine ratio, as well as the pancreas and renal structural disturbance. Glomerular filtration rate, BP and HR increased in the DN group. The ACE activity was elevated in the serum and kidneys of the DN group. Administration of hWJMSCs-CM modulated the renal functional and structural disturbance and decreased the ACE activity. Conclusion: Conditioned media was extracted from hWJMSCs may have a Renoprotective effect in diabetic nephropathy. This may happen through regulation of ACE activity and renin-angiotensin system inhibition.

Keywords: diabetic nephropathy, mesenchymal stem cells, immunomodulation, anti-inflammation

Procedia PDF Downloads 175

Authors: Arjun Dakuri, J. Hayavadana

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The study includes selection of 100 % bamboo fabric and cotton fabric for the study. The 100% bamboo fabrics were of 127 g/m², and 112 g/m² and 100% cotton grey fabric were of 104 g/m². The cotton fabric was desized, scoured, bleached and then treated with ZnO (as antimicrobial agent) with 1%, 2% and 3% using pad-dry cure method, whereas the bamboo fabrics were only desized. The antimicrobial activity of bamboo and ZnO treated cotton fabrics were evaluated and compared against E. coli and S. aureus as per the standard AATCC - 147. Moisture management properties of selected fabrics were also analyzed. Further, the selected fabric samples were tested for comfort properties like bending length, tearing strength, drape-ability, and specific handle force and air permeability. It was observed that bamboo fabrics show significant antibacterial activity and the same was shown by 3% ZnO treated cotton fabric. Both cotton and bamboo fabrics show improved moisture management properties than the cotton fabric. The comfort properties of bamboo fabrics are found to be superior to cotton fabrics making it more suitable for applications in place of cotton.

Keywords: antimicrobial activity, bamboo, cotton, comfort properties, moisture management, zinc oxide

Procedia PDF Downloads 318

Authors: Wen-Shyong Tzou, Chih-Ching Huang, Chin-Hwa Hu, Ying-Tsang Lo, Tun-Wen Pai, Chia-Yin Chiang, Chung-Hao Li, Hong-Jyuan Jian

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Plasmin plays an important role in the human circulatory system owing to its catalytic ability of fibrinolysis. The immediate injection of plasmin in patients of strokes has intrigued many scientists to design vectors that can transport plasmin to the desired location in human body. Here we predict the structure of human plasmin and investigate the interaction of plasmin with the gold-nanoparticle. Because the crystal structure of plasminogen has been solved, we deleted N-terminal domain (Pan-apple domain) of plasminogen and generate a mimic of the active form of this enzyme (plasmin). We conducted a simulated annealing process on plasmin and discovered a very large conformation occurs. Kringle domains 1, 4 and 5 had been observed to leave its original location relative to the main body of the enzyme and the original doughnut shape of this enzyme has been transformed to a V-shaped by opening its two arms. This observation of conformational change is consistent with the experimental results of neutron scattering and centrifugation. We subsequently docked the plasmin on the simulated gold surface to predict their interaction. The V-shaped plasmin could utilize its Kringle domain and catalytic domain to contact the gold surface. Our findings not only reveal the flexibility of plasmin structure but also provide a guide for the design of a plasmin-gold nanoparticle.

Keywords: docking, gold nanoparticle, molecular simulation, plasmin

Procedia PDF Downloads 450