International Science Index

International Journal of Biotechnology and Bioengineering

482
58864
Potential Activities of Human Endogenous Retroviral kDNA in Melanoma Pathogenesis and HIV-1 Infection
Abstract:
Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.
481
56779
Sorbitol Galactoside Synthesis Using β-Galactosidase Immobilized on Functionalized Silica Nanoparticles
Abstract:
Nowadays, considering the growing awareness of functional food beneficial effects on human health, due attention is dedicated to the research in the field of obtaining new prominent products exhibiting improved physiological and physicochemical characteristics. Therefore, different approaches to valuable bioactive compounds synthesis have been proposed. β-Galactosidase, for example, although mainly utilized as hydrolytic enzyme, proved to be a promising tool for these purposes. Namely, under the particular conditions, such as high lactose concentration, elevated temperatures and low water activities, reaction of galactose moiety transfer to free hydroxyl group of the alternative acceptor (e.g. different sugars, alcohols or aromatic compounds) can generate a wide range of potentially interesting products. Up to now, galacto-oligosaccharides and lactulose have attracted the most attention due to their inherent prebiotic properties. The goal of this study was to obtain a novel product sorbitol galactoside, using the similar reaction mechanism, namely transgalactosylation reaction catalyzed by β-galactosidase from Aspergillus oryzae. By using sugar alcohol (sorbitol) as alternative acceptor, a diverse mixture of potential prebiotics is produced, enabling its more favorable functional features. Nevertheless, an introduction of alternative acceptor into the reaction mixture contributed to the complexity of reaction scheme, since several potential reaction pathways were introduced. Therefore, the thorough optimization using response surface method (RSM), in order to get an insight into different parameter (lactose concentration, sorbitol to lactose molar ratio, enzyme concentration, NaCl concentration and reaction time) influences, as well as their mutual interactions on product yield and productivity, was performed. In view of product yield maximization, the obtained model predicted optimal lactose concentration 500 mM, the molar ratio of sobitol to lactose 9, enzyme concentration 0.76 mg/ml, concentration of NaCl 0.8M, and the reaction time 7h. From the aspect of productivity, the optimum substrate molar ratio was found to be 1, while the values for other factors coincide. In order to additionally, improve enzyme efficiency and enable its reuse and potential continual application, immobilization of β-galactosidase onto tailored silica nanoparticles was performed. These non-porous fumed silica nanoparticles (FNS)were chosen on the basis of their biocompatibility and non-toxicity, as well as their advantageous mechanical and hydrodinamical properties. However, in order to achieve better compatibility between enzymes and the carrier, modifications of the silica surface using amino functional organosilane (3-aminopropyltrimethoxysilane, APTMS) were made. Obtained support with amino functional groups (AFNS) enabled high enzyme loadings and, more importantly, extremely high expressed activities, approximately 230 mg proteins/g and 2100 IU/g, respectively. Moreover, this immobilized preparation showed high affinity towards sorbitol galactoside synthesis. Therefore, the findings of this study could provided a valuable contribution to the efficient production of physiologically active galactosides in immobilized enzyme reactors.
480
59614
Purification of Bacterial Lipopeptide Antimicrobials Effective against Tuberculosis
Abstract:
Lipopeptides are microbially-synthesized surfactants that find application in pharmaceutical, agricultural, food, bioremediation and cosmetic industries. Specifically, the potential for the Bacillus surfactin lipopeptide as an alternative antibacterial agent effective against the causative agent of tuberculosis (TB), Mycobacterium tuberculosis, has prompted the systematic investigation of upstream and downstream processing presented in this study. It is envisaged that this research will lead to improved bioprocessing economy of the surfactin lipopeptide for use as a disinfecting agent against tuberculosis, thereby reducing the deaths due to TB globally. The study investigates the kinetics of Bacillus subtilis ATCC 21332 with respect to biomass growth and its associated lipopeptide production in both shake-flask and bioreactor culture, and extends to develop a purification procedure for optimal recovery and purification of surfactin from the culture supernatant. The production studies were performed in a glucose mineral salt medium optimised according to nitrogen source and dissolved oxygen availability. Lipopeptides were recovered from the culture supernatant by acid precipitation at pH 2 using 4 N HCl. The acid precipitate containing lipopeptides was further purified by extraction using the solvents methanol, acetonitrile, acetone, propanol, chloroform, petroleum ether, hexane, tert-butyl-methy ether and ethyl acetate. The concentrations of surfactin recovered in the acid precipitates and the solvent extracts, as well as the purities with respect to surfactin, were determined using thin layer chromatography, with results further validated by reverse phase-high performance liquid chromatography. Finally, the antimicrobial efficacy of the purified lipopeptides was tested against M. aurum, a GRAS surrogate of M. tuberculosis. The nitrogen source and dissolved oxygen availability showed significant effect on the surfactin production thereby facilitating the quantification of upstream conditions for optimal surfactin production. The subsequent purification of the lipopeptide–containing supernatant by acid precipitation yielded about 95 % of surfactin recovery with a purity of approx. 50%. Subsequent solvent extraction of the resolubilised acid precipitate showed varying recoveries from 5±1% in petroleum ether to 100±8% in methanol, with % purity varying upwards of 48±10% (in a chloroform:methanol=50:50 mixture) in the different solvents. These studies informed on a two-staged solvent extraction procedure to improve the overall purity of surfactin whilst maximising its recovery. For instance, solvent extraction using petroleum ether could successfully remove the lipid-impurities in the first stage, followed by extraction with methanol to improve the overall purity. Using this or similar purification strategies, it is envisaged to produce purified surfactin suitable for a wide range of disinfection applications. While these studies have specific significance for tuberculosis disinfection, they also have broad-based generic implications for other antimicrobial applications of lipopeptides. Of particular importance in this context is the antifungicidal efficacy of the coproduced iturin and fengycin against agricultural disease. In this case, analogous purification studies showed a range of recovery of these anti-fungicidal lipopeptides from 16±1% to 100±0% using octanol and methanol as solvents respectively, whilst a maximum of 64±12% purity was obtained in isopropanol.
479
56326
Introduction of Electronic Health Records to Improve Data Quality in Emergency Department Operations
Abstract:
In its simplest form, data quality can be defined as 'fitness for use' and it is a concept with multi-dimensions. Emergency Departments (ED) require information to treat patients and on the other hand it is the primary source of information regarding accidents, injuries, emergencies etc. Also, it is the starting point of various patient registries, databases and surveillance systems. This interventional study was carried out to improve data quality at the ED of the National Hospital of Sri Lanka (NHSL) by introducing an e-health solution to improve data quality. The NHSL is the premier trauma care centre in Sri Lanka. The study consisted of three components. A research study was conducted to assess quality of data in relation to selected five dimensions of data quality namely accuracy, completeness, timeliness, legibility and reliability. The intervention was to develop and deploy an electronic emergency department information system (eEDIS). Post assessment of the intervention confirmed that all five dimensions of data quality had improved. The most significant improvement noticed in accuracy and timeliness dimensions.
478
56328
Introduction of Digital Radiology to Improve the Timeliness in Availability of Radiological Diagnostic Images for Trauma Care
Abstract:
In an emergency department ‘where every second counts for patient’s management’ timely availability of X- rays play a vital role in early diagnosis and management of patients. Trauma care centers rely heavily on timely radiologic imaging for patient care and radiology plays a crucial role in the emergency department (ED) operations. A research study was carried out to assess timeliness of availability of X-rays and total turnaround time at the Accident Service of National Hospital of Sri Lanka which is the premier trauma centre in the country. Digital Radiology system was implemented as an intervention to improve the timeliness of availability of X-rays. Post-implementation assessment was carried out to assess the effectiveness of the intervention. Reduction in all three aspects of waiting times namely waiting for initial examination by doctors, waiting until X –ray is performed and waiting for image availability was observed after implementation of the intervention. However, the most significant improvement was seen in waiting time for image availability and reduction in time for image availability had indirect impact on reducing waiting time for initial examination by doctors and waiting until X –ray is performed. The most significant reduction in time for image availability was observed when performing 4-5 X-rays with DR system. The least improvement in timeliness was seen in patients who categorized as critical.
477
59932
Development of Recombinant Bacillus subtilis Strain for the Production of 3-Hydroxypropionic Acid
Abstract:
3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. Recombinant strains containing the heterologous genes without any codon optimization failed to produce any 3-HP. By contrast, the recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Knocking out the glycerol kinase (glpk), combined with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the successful report of the introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.
476
48916
Exploitation of Terpenes as Guardians in Plant Biotechnology
Abstract:
Plants are always being threatened by biotic and abiotic elements in their abode. Although they have inherited mechanisms to defend themselves, sometimes due to overpowering of their enemies or weakening of themselves, they just suffer from those elements. Human, as to help plants defend themselves, have developed several methods among which application of terpenes via plant biotechnology is promising. Terpenes are the most frequent and diverse secondary metabolites in plants. In these plants, terpenes are involved in different protective aspects. In this field, by utilizing biotechnological approaches on them, a delicate, precise, and an economic intervention will be achieved. In this review, first, the importance of terpenes as guardians in plants, which include their allelopathy effect, a call for alliances, and a mitigation impact on abiotic stresses will be pointed out. Second, problems concerning terpenes application in plant biotechnology comprising: damage to cell, undesirable terpene production and undesirable concentration and proportion of terpenes will be discussed. At the end, the approaches in plant biotechnology of terpenes including tampering with terpene gene sequences, compartmentalization, and localization and utilization of membrane transporters will be expressed. It is concluded with some useful notions concerning the topic.
475
56843
Microbial Fuel Cells and Their Applications in Electricity Generating and Wastewater Treatment
Authors:
Abstract:
This research is an experimental research which was done about microbial fuel cells in order to study them for electricity generating and wastewater treatment. These days it is very important to find new, clean and sustainable ways for energy supplying. Because of this reason there are many researchers around the world who are studying about new and sustainable energies. There are different ways to produce these kind of energies like: solar cells, wind turbines, geothermal energy, fuel cells and many other ways. Fuel cells have different types one of these types is microbial fuel cell. In this research a MFC was built in order to study how it can be used for electricity generating and wastewater treatment. The microbial fuel cell which was used in this research is a reactor that has two tanks with a catalyst solution. The chemical reaction in microbial fuel cells is a redox reaction. This paragraph explains the methodology and materials. The microbial fuel cell in this research is a two chamber MFC. Anode chamber is an anaerobic one (ABR reactor) and the other chamber is a cathode chamber. Anode chamber consists of stabilized sludge which is the source of microorganisms that do redox reaction. The main microorganisms here are: Propionibacterium and Clostridium. The electrodes of anode chamber are graphite pages. Cathode chamber consists of graphite page electrodes and catalysts like: O2, KMnO4 and C6N6FeK4. The membrane which separates the chambers is Nafion117. The reason of choosing this membrane is explained in the complete paper. The main goal of this research as it is clear in the topic is to generate electricity and treating wastewater. Some results and conclusions which were found: it was found that when you use electron receptor compounds like: O2 , MnO4 , C6N6FeK4 the velocity of electron receiving speeds up and in a less time more current will be achieved . It was found that the best compounds for this purpose are compounds which have iron in their chemical formula. It is also important to pay attention to the amount of nutrition which enters to bacteria chamber. By adding extra nutrition in some cases the result will be reverse. By using ABR the amount of chemical oxidation demand reduces per day till it arrives to a stable amount.
474
59776
Industrial and Technological Applications of Brewer’s Spent Malt
Abstract:
During industrial processing of raw materials of animal and vegetable origin, large amounts of solid, liquid and gaseous wastes are generated. Solid residues are usually materials rich in carbohydrates, protein, fiber and minerals. Brewer’s spent grain (BSG) is the main waste generated in the brewing industry, representing 85% of the waste generated in this industry. It is estimated that world’s BSG generation is approximately 38.6 x 106 t per year and represents 20-30% (w/w) of the initial mass of added malt, resulting in low commercial value by-product, however, does not have economic value, but it must be removed from the brewery, as its spontaneous fermentation can attract insects and rodents. For every 100 grams in dry basis, BSG has approximately 68 g total fiber, being divided into 3.5 g of soluble fiber and 64.3 g of insoluble fiber (cellulose, hemicellulose and lignin). In addition to dietary fibers, depending on the efficiency of the grinding process and mashing, BSG may also have starch, reducing sugars, lipids, phenolics and antioxidants, emphasizing that its composition will depend on the barley variety and cultivation conditions, malting and technology involved in the production of beer. BSG demands space for storage, but studies have proposed alternatives such as the use of drying, extrusion, pressing with superheated steam, and grinding to facilitate storage. Other important characteristics that enhance its applicability in bioremediation, effluent treatment and biotechnology, is the surface area (SBET) of 1.748 m2 g-1, total pore volume of 0.0053 cm3 g-1 and mean pore diameter of 121.784 Å, characterized as a macroporous and possess fewer adsorption properties but have great ability to trap suspended solids for separation from liquid solutions. It has low economic value; however, it has enormous potential for technological applications that can improve or add value to this agro-industrial waste. Due to its composition, this material has been used in several industrial applications such as in the production of food ingredients, fiber enrichment by its addition in foods such as breads and cookies in bioremediation processes, substrate for microorganism and production of biomolecules, bioenergy generation, and civil construction, among others. Therefore, the use of this waste or by-product becomes essential and aimed at reducing the amount of organic waste in different industrial processes, especially in breweries.
473
59776
Industrial and Technological Applications of Brewer’s Spent Malt
Abstract:
During industrial processing of raw materials of animal and vegetable origin, large amounts of solid, liquid and gaseous wastes are generated. Solid residues are usually materials rich in carbohydrates, protein, fiber and minerals. Brewer’s spent grain (BSG) is the main waste generated in the brewing industry, representing 85% of the waste generated in this industry. It is estimated that world’s BSG generation is approximately 38.6 x 106 t per year and represents 20-30% (w/w) of the initial mass of added malt, resulting in low commercial value by-product, however, does not have economic value, but it must be removed from the brewery, as its spontaneous fermentation can attract insects and rodents. For every 100 grams in dry basis, BSG has approximately 68 g total fiber, being divided into 3.5 g of soluble fiber and 64.3 g of insoluble fiber (cellulose, hemicellulose and lignin). In addition to dietary fibers, depending on the efficiency of the grinding process and mashing, BSG may also have starch, reducing sugars, lipids, phenolics and antioxidants, emphasizing that its composition will depend on the barley variety and cultivation conditions, malting and technology involved in the production of beer. BSG demands space for storage, but studies have proposed alternatives such as the use of drying, extrusion, pressing with superheated steam, and grinding to facilitate storage. Other important characteristics that enhance its applicability in bioremediation, effluent treatment and biotechnology, is the surface area (SBET) of 1.748 m2 g-1, total pore volume of 0.0053 cm3 g-1 and mean pore diameter of 121.784 Å, characterized as a macroporous and possess fewer adsorption properties but have great ability to trap suspended solids for separation from liquid solutions. It has low economic value; however, it has enormous potential for technological applications that can improve or add value to this agro-industrial waste. Due to its composition, this material has been used in several industrial applications such as in the production of food ingredients, fiber enrichment by its addition in foods such as breads and cookies in bioremediation processes, substrate for microorganism and production of biomolecules, bioenergy generation, and civil construction, among others. Therefore, the use of this waste or by-product becomes essential and aimed at reducing the amount of organic waste in different industrial processes, especially in breweries.
472
59150
Chemical Synthesis of a cDNA and Its Expression Analysis
Abstract:
Synthetic cDNA (ScDNA) of granulocyte colony-stimulating factor (G-CSF) was constructed using a DNA synthesizer with the aim to increase its expression level. 5' end of the ScDNA of G-CSF coding region was modified by decreasing the GC content without altering the predicted amino acids sequence. The identity of the resulting protein from ScDNA was confirmed by the highly specific enzyme-linked immunosorbent assay. In conclusion, a synthetic G-CSF cDNA in combination with the recombinant DNA protocol offers a rapid and reliable strategy for synthesizing the target protein. However, the commercial utilization of this methodology requires rigorous validation and quality control.
471
58645
Cell Microarrays for the Design of Biomaterials to Support Human Endothelial Cell Growth
Abstract:
Endothelial progenitor cell (EPCs) based-therapies continue to be of interest for administration to treat ischemic events rather than organ and tissue transplantation due to the central role EPCs play in blood vessel formation and tissue re-vascularisation. One of the current strategies for the production of clinical-grade EPCs requires the in vitro isolation and proliferation of EPCs from peripheral blood samples to provide sufficient quantities of functional EPCs for further cell therapies. This study aims to examine the interaction of EPCs with different biomolecules to significantly improve the current protocols/strategies for EPC capture and expansion while maintaining their endothelial-like nature and functional capabilities. Readout assays including the uptake of acetylated low-density lipoprotein (ac-LDL) and binding of Ulex europaeus agglutinin (UEA-1) were performed to confirm the cells were viable and functional after 3 days culture on immobilised biomolecules. In this study, four different biomolecules were investigated for their capacity to support EPC capture and proliferation when immobilised on a surface. For reasons of confidentiality all printed biomolecules have been de-identified as biomolecules “a”, “b”, “c”, and “d” respectively. Firstly, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules were then arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were cultured on the microarray platform and cell numbers quantified after 1 h (to determine capture) and 3 days (to determine proliferation). All the printed biomolecules showed the ability to capture EPCs within 1 h of cell seeding with the least number of EPCs binding to immobilised biomolecule “d”. Among the first biomolecules, biomolecule ”b” exhibited the highest EPC attachment after 1 h in comparison to biomolecule ”a” and biomolecule ”c” which are averaged 31, 20 and 17 cells per spot, respectively. On the other hand, biomolecule ”c” exhibited a significant increase in EPC expansion after 3 days when compared to biomolecule ”a” and biomolecule ”b” with a 4-fold increase in cell number. In addition, the functionality of expanded EPCs on these biomolecules was successfully verified via uptake of ac-LDL and binding of UEA-1. These results provide a new and significant improvement in the capture and expansion of human EPCs, which provide advances in the application of autologous and/or allogeneic EPCs for cell therapy in several debilitating and life-threatening diseases.
470
57656
Nanomechanical Characterization of Healthy and Tumor Lung Tissues at Cell and Extracellular Matrix Level
Abstract:
The study of the biophysics of living cells drew attention to the pivotal role of the cytoskeleton in many cell functions, such as mechanics, adhesion, proliferation, migration, differentiation and neoplastic transformation. In particular, during the complex process of malignant transformation and invasion cell cytoskeleton devolves from a rigid and organized structure to a more compliant state, which confers to the cancer cells a great ability to migrate and adapt to the extracellular environment. In order to better understand the malignant transformation process from a mechanical point of view, it is necessary to evaluate the direct crosstalk between the cells and their surrounding extracellular matrix (ECM) in a context which is close to in vivo conditions. In this study, human biopsy tissues of lung adenocarcinoma were analyzed in order to define their mechanical phenotype at cell and ECM level, by using particle tracking microrheology (PTM) technique. Polystyrene beads (500 nm) were introduced into the sample slice. The motion of beads was obtained by tracking their displacements across cell cytoskeleton and ECM structures and mean squared displacements (MSDs) were calculated from bead trajectories. It has been already demonstrated that the amplitude of MSD is inversely related to the mechanical properties of intracellular and extracellular microenvironment. For this reason, MSDs of particles introduced in cytoplasm and ECM of healthy and tumor tissues were compared. PTM analyses showed that cancerous transformation compromises mechanical integrity of cells and extracellular matrix. In particular, the MSD amplitudes in cells of adenocarcinoma were greater as compared to cells of normal tissues. The increased motion is probably associated to a less structured cytoskeleton and consequently to an increase of deformability of cells. Further, cancer transformation is also accompanied by extracellular matrix stiffening, as confirmed by the decrease of MSDs of matrix in tumor tissue, a process that promotes tumor proliferation and invasiveness, by activating typical oncogenic signaling pathways. In addition, a clear correlation between MSDs of cells and tumor grade was found. MSDs increase when tumor grade passes from 2 to 3, indicating that cells undergo to a trans-differentiation process during tumor progression. ECM stiffening is not dependent on tumor grade, but the tumor stage resulted to be strictly correlated with both cells and ECM mechanical properties. In fact, a greater stage is assigned to tumor spread to regional lymph nodes and characterized by an up-regulation of different ECM proteins, such as collagen I fibers. These results indicate that PTM can be used to get nanomechanical characterization at different scale levels in an interpretative and diagnostic context.
469
58283
Effects of Zeolite and Zinc on Quality of Canola (Brassica napus L.) under Late Season Drought Stress
Abstract:
Canola (Brassica napus L.) is one of the most important oil crops severely affected by drought. Drought stress, due to low precipitation and soil water reserve, has resulted in severe decline in crop yield in Iran. The decline in yield is aggravated by zinc deficiency in the soil and the lack of drought stress tolerant cultivars. In Iran, soil alkalinity caused widespread zinc deficiencies which severely affect canola planted under adequate fertilizer regime. This study was conducted between 2010 to 2012 growing seasons to investigate the effect of zeolite and foliar zinc application on growth, yield and yield components of three (3) canola cultivars at two (2) irrigation regimes. The research was conducted using factorial split-plot experiment based on the randomized complete block design (RCBD) with three replications. The treatments used in the study including: 1) irrigation (I): complete (normal irrigation) (I1), and restricted irrigation (I2) applied at the pod formation stage, 2) zeolite (Z): 0 (Z1), and 15 t ha-1 (Z2), and 3) Zn: zinc sulphate concentrations of 0%, 0.1%, and 0.2 % (Zn1, Zn2, and Zn3) on Opera, Licord, and RGS003 cultivars. The results showed that the simple effects of treatments were highly significant for all characteristics at P < 0.01. Normal irrigation, application of zeolite, and zinc had a positive effect on the oil content, and oil yield. The combined application of zeolite and zinc decreased the yield and yield components, oil content and oil yield were obtained by applying Z2Zn2 (15 t ha-1 zeolite and 0.1% zinc sulphate) in both irrigation regimes. Opera cultivar in normal irrigation and RGS003 cultivar in stress conditions had the highest yield and yield components, oil yield, and oil content. Iran needs more than one solution to improve oil yield of canola planted during late season drought stress. Therefore, based on the low cost of natural zeolite and moderate dose application of zinc, these treatments are recommended to enhance the growth of canola in regions prone to drought stress. Other economical plants of Iran such as wheat, saffron and pistachio are expected to benefit from the findings from this study.
Keywords:
468
58283
Effects of Zeolite and Zinc on Quality of Canola (Brassica napus L.) under Late Season Drought Stress
Abstract:
Canola (Brassica napus L.) is one of the most important oil crops severely affected by drought. Drought stress, due to low precipitation and soil water reserve, has resulted in severe decline in crop yield in Iran. The decline in yield is aggravated by zinc deficiency in the soil and the lack of drought stress tolerant cultivars. In Iran, soil alkalinity caused widespread zinc deficiencies which severely affect canola planted under adequate fertilizer regime. This study was conducted between 2010 to 2012 growing seasons to investigate the effect of zeolite and foliar zinc application on growth, yield and yield components of three (3) canola cultivars at two (2) irrigation regimes. The research was conducted using factorial split-plot experiment based on the randomized complete block design (RCBD) with three replications. The treatments used in the study including: 1) irrigation (I): complete (normal irrigation) (I1), and restricted irrigation (I2) applied at the pod formation stage, 2) zeolite (Z): 0 (Z1), and 15 t ha-1 (Z2), and 3) Zn: zinc sulphate concentrations of 0%, 0.1%, and 0.2 % (Zn1, Zn2, and Zn3) on Opera, Licord, and RGS003 cultivars. The results showed that the simple effects of treatments were highly significant for all characteristics at P < 0.01. Normal irrigation, application of zeolite, and zinc had a positive effect on the oil content, and oil yield. The combined application of zeolite and zinc decreased the yield and yield components, oil content and oil yield were obtained by applying Z2Zn2 (15 t ha-1 zeolite and 0.1% zinc sulphate) in both irrigation regimes. Opera cultivar in normal irrigation and RGS003 cultivar in stress conditions had the highest yield and yield components, oil yield, and oil content. Iran needs more than one solution to improve oil yield of canola planted during late season drought stress. Therefore, based on the low cost of natural zeolite and moderate dose application of zinc, these treatments are recommended to enhance the growth of canola in regions prone to drought stress. Other economical plants of Iran such as wheat, saffron and pistachio are expected to benefit from the findings from this study.
Keywords:
467
58283
Effects of Zeolite and Zinc on Quality of Canola (Brassica napus L.) under Late Season Drought Stress
Abstract:
Canola (Brassica napus L.) is one of the most important oil crops severely affected by drought. Drought stress, due to low precipitation and soil water reserve, has resulted in severe decline in crop yield in Iran. The decline in yield is aggravated by zinc deficiency in the soil and the lack of drought stress tolerant cultivars. In Iran, soil alkalinity caused widespread zinc deficiencies which severely affect canola planted under adequate fertilizer regime. This study was conducted between 2010 to 2012 growing seasons to investigate the effect of zeolite and foliar zinc application on growth, yield and yield components of three (3) canola cultivars at two (2) irrigation regimes. The research was conducted using factorial split-plot experiment based on the randomized complete block design (RCBD) with three replications. The treatments used in the study including: 1) irrigation (I): complete (normal irrigation) (I1), and restricted irrigation (I2) applied at the pod formation stage, 2) zeolite (Z): 0 (Z1), and 15 t ha-1 (Z2), and 3) Zn: zinc sulphate concentrations of 0%, 0.1%, and 0.2 % (Zn1, Zn2, and Zn3) on Opera, Licord, and RGS003 cultivars. The results showed that the simple effects of treatments were highly significant for all characteristics at P < 0.01. Normal irrigation, application of zeolite, and zinc had a positive effect on the oil content, and oil yield. The combined application of zeolite and zinc decreased the yield and yield components, oil content and oil yield were obtained by applying Z2Zn2 (15 t ha-1 zeolite and 0.1% zinc sulphate) in both irrigation regimes. Opera cultivar in normal irrigation and RGS003 cultivar in stress conditions had the highest yield and yield components, oil yield, and oil content. Iran needs more than one solution to improve oil yield of canola planted during late season drought stress. Therefore, based on the low cost of natural zeolite and moderate dose application of zinc, these treatments are recommended to enhance the growth of canola in regions prone to drought stress. Other economical plants of Iran such as wheat, saffron and pistachio are expected to benefit from the findings from this study.
Keywords:
466
54893
Anaerobic Fermentation Process for Production of Biohydrogen from Pretreated Fruit Wastes
Abstract:
Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.
465
54893
Anaerobic Fermentation Process for Production of Biohydrogen from Pretreated Fruit Wastes
Abstract:
Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.
464
54893
Anaerobic Fermentation Process for Production of Biohydrogen from Pretreated Fruit Wastes
Abstract:
Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.
463
54893
Anaerobic Fermentation Process for Production of Biohydrogen from Pretreated Fruit Wastes
Abstract:
Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.
462
54893
Anaerobic Fermentation Process for Production of Biohydrogen from Pretreated Fruit Wastes
Abstract:
Fruit waste was used as a feedstock to produce biohydrogen in this study. Fruit waste used in this study was collected from several fruit juice stalls in Singapore. Based on our observation, the fruit waste contained 35-40% orange, 10-20% watermelon, 10-15% apple, 10-15% pineapple, 1-5% mango. They were mixed with water (1:1 ratio based on wet biomass) and blended to attain homogenous mixtures. Later, fruit waste was subjected to one of the following pretreatments: autoclave (121 °C for 20min), microwave (20min) or both. After pretreatment, the total sugar concentration in the hydrolysate was high (>12g/l) when both autoclave and microwave were applied. In contrast, samples without pretreatment measured only less than 2g/l of sugar. While using these hydrolysates as carbon sources, Clostridium strain BOH3 produces 2526-3126 ml/l of hydrogen after 72h of anaerobic fermentation. The hydrogen yield was 295-300 ml/g of sugar which is close to the hydrogen yields from glucose (338 ml/gm) and xylose (330 ml/gm). Our HPLC analysis showed that fruit waste hydrolysate contained oligosugars (25-27%), sucrose (18-23%), fructose (25-30%), glucose (10-15%) and mannose (2-5%). Additionally, pretreatment led to the release of free amino acids (160-512 mg/l), calcium (7.8-12.9 ppm), magnesium (4.32-6.55 ppm), potassium (5.4-65.1 ppm) and sodium (0.4-0.5 ppm) into the hydrolysate. These nutrients were able to support strain-BOH3 to grow and produce high level of hydrogen. Notably, unlike other pretreatment methods (with strong acids and bases), these pretreatment techniques did not generate any inhibitors (e.g. furfural and phenolic acids) to suppress the hydrogen production. Interestingly, strain BOH3 can also ferment pretreated fruit waste slurry and produce hydrogen with a high yield (156-343 ml/gm fruit waste). While fermenting pretreated fruit waste slurry, strain-BOH3 excreted several saccharolytic enzymes majorly xylanase (1.84U/ml), amylase (1.10U/ml), pectinase (0.36U/ml) and cellulase (0.43U/ml). Due to expressions of these enzymes, strain BOH3 was able to directly utilize pretreated fruit waste hydrolysate and produces high-level of hydrogen.
461
54915
Chitosan Doped Curcumin Gold Clusters Flexible Nanofiber for Wound Dressing and Anticancer Activities
Abstract:
The purpose of this study is to develop the chitosan doped curcumin gold cluster nanofiber for wound healing and skin cancer drug delivery applications. Chitosan is a typical marine polysaccharide composed of glucosamine and n-acetyl glucosamine biodegradable and biocompatible polymer. Curcumin is a natural bioactive molecule obtained from Curcuma longo, it mostly occurs in some Asian countries like India and China. It has naturally antioxidant, antimicrobial, wound healing and anticancer property. Due to this advantage, we prepared a combination of natural polymer chitosan with Curcumin and gold nanocluster nanofiber (CH-CUR-AuNCs nanofibers). The prepared nanofiber was characterized by using Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Antibacterial studies were performed with E.coli and S.aureus. Antioxidant assay, drug release test, and cytotoxicity will be evaluated. Prepared nanofiber emits low intensity of red fluorescent. The FTIR confirm the presence of chitosan and Curcumin in the nanofiber. In vitro study clearly shows the antibacterial activity against the gram negative and gram positive bacteria. Particularly, synthesised nanofibers provide better antibacterial activity against gram negative than gram positive. Cytotoxicity study also provides better killing rate in cancer cell, biocompatible with normal cell. Prepared CH-CUR-AuNCs nanofibers provide the better killing rate to bacterial strains and cancer cells. Finally, prepared nanofiber can be possible to use for wound healing dressing, patch for skin cancer and other biomedical applications.
460
54915
Chitosan Doped Curcumin Gold Clusters Flexible Nanofiber for Wound Dressing and Anticancer Activities
Abstract:
The purpose of this study is to develop the chitosan doped curcumin gold cluster nanofiber for wound healing and skin cancer drug delivery applications. Chitosan is a typical marine polysaccharide composed of glucosamine and n-acetyl glucosamine biodegradable and biocompatible polymer. Curcumin is a natural bioactive molecule obtained from Curcuma longo, it mostly occurs in some Asian countries like India and China. It has naturally antioxidant, antimicrobial, wound healing and anticancer property. Due to this advantage, we prepared a combination of natural polymer chitosan with Curcumin and gold nanocluster nanofiber (CH-CUR-AuNCs nanofibers). The prepared nanofiber was characterized by using Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Antibacterial studies were performed with E.coli and S.aureus. Antioxidant assay, drug release test, and cytotoxicity will be evaluated. Prepared nanofiber emits low intensity of red fluorescent. The FTIR confirm the presence of chitosan and Curcumin in the nanofiber. In vitro study clearly shows the antibacterial activity against the gram negative and gram positive bacteria. Particularly, synthesised nanofibers provide better antibacterial activity against gram negative than gram positive. Cytotoxicity study also provides better killing rate in cancer cell, biocompatible with normal cell. Prepared CH-CUR-AuNCs nanofibers provide the better killing rate to bacterial strains and cancer cells. Finally, prepared nanofiber can be possible to use for wound healing dressing, patch for skin cancer and other biomedical applications.
459
54915
Chitosan Doped Curcumin Gold Clusters Flexible Nanofiber for Wound Dressing and Anticancer Activities
Abstract:
The purpose of this study is to develop the chitosan doped curcumin gold cluster nanofiber for wound healing and skin cancer drug delivery applications. Chitosan is a typical marine polysaccharide composed of glucosamine and n-acetyl glucosamine biodegradable and biocompatible polymer. Curcumin is a natural bioactive molecule obtained from Curcuma longo, it mostly occurs in some Asian countries like India and China. It has naturally antioxidant, antimicrobial, wound healing and anticancer property. Due to this advantage, we prepared a combination of natural polymer chitosan with Curcumin and gold nanocluster nanofiber (CH-CUR-AuNCs nanofibers). The prepared nanofiber was characterized by using Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Antibacterial studies were performed with E.coli and S.aureus. Antioxidant assay, drug release test, and cytotoxicity will be evaluated. Prepared nanofiber emits low intensity of red fluorescent. The FTIR confirm the presence of chitosan and Curcumin in the nanofiber. In vitro study clearly shows the antibacterial activity against the gram negative and gram positive bacteria. Particularly, synthesised nanofibers provide better antibacterial activity against gram negative than gram positive. Cytotoxicity study also provides better killing rate in cancer cell, biocompatible with normal cell. Prepared CH-CUR-AuNCs nanofibers provide the better killing rate to bacterial strains and cancer cells. Finally, prepared nanofiber can be possible to use for wound healing dressing, patch for skin cancer and other biomedical applications.
458
54915
Chitosan Doped Curcumin Gold Clusters Flexible Nanofiber for Wound Dressing and Anticancer Activities
Abstract:
The purpose of this study is to develop the chitosan doped curcumin gold cluster nanofiber for wound healing and skin cancer drug delivery applications. Chitosan is a typical marine polysaccharide composed of glucosamine and n-acetyl glucosamine biodegradable and biocompatible polymer. Curcumin is a natural bioactive molecule obtained from Curcuma longo, it mostly occurs in some Asian countries like India and China. It has naturally antioxidant, antimicrobial, wound healing and anticancer property. Due to this advantage, we prepared a combination of natural polymer chitosan with Curcumin and gold nanocluster nanofiber (CH-CUR-AuNCs nanofibers). The prepared nanofiber was characterized by using Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Antibacterial studies were performed with E.coli and S.aureus. Antioxidant assay, drug release test, and cytotoxicity will be evaluated. Prepared nanofiber emits low intensity of red fluorescent. The FTIR confirm the presence of chitosan and Curcumin in the nanofiber. In vitro study clearly shows the antibacterial activity against the gram negative and gram positive bacteria. Particularly, synthesised nanofibers provide better antibacterial activity against gram negative than gram positive. Cytotoxicity study also provides better killing rate in cancer cell, biocompatible with normal cell. Prepared CH-CUR-AuNCs nanofibers provide the better killing rate to bacterial strains and cancer cells. Finally, prepared nanofiber can be possible to use for wound healing dressing, patch for skin cancer and other biomedical applications.
457
54915
Chitosan Doped Curcumin Gold Clusters Flexible Nanofiber for Wound Dressing and Anticancer Activities
Abstract:
The purpose of this study is to develop the chitosan doped curcumin gold cluster nanofiber for wound healing and skin cancer drug delivery applications. Chitosan is a typical marine polysaccharide composed of glucosamine and n-acetyl glucosamine biodegradable and biocompatible polymer. Curcumin is a natural bioactive molecule obtained from Curcuma longo, it mostly occurs in some Asian countries like India and China. It has naturally antioxidant, antimicrobial, wound healing and anticancer property. Due to this advantage, we prepared a combination of natural polymer chitosan with Curcumin and gold nanocluster nanofiber (CH-CUR-AuNCs nanofibers). The prepared nanofiber was characterized by using Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Antibacterial studies were performed with E.coli and S.aureus. Antioxidant assay, drug release test, and cytotoxicity will be evaluated. Prepared nanofiber emits low intensity of red fluorescent. The FTIR confirm the presence of chitosan and Curcumin in the nanofiber. In vitro study clearly shows the antibacterial activity against the gram negative and gram positive bacteria. Particularly, synthesised nanofibers provide better antibacterial activity against gram negative than gram positive. Cytotoxicity study also provides better killing rate in cancer cell, biocompatible with normal cell. Prepared CH-CUR-AuNCs nanofibers provide the better killing rate to bacterial strains and cancer cells. Finally, prepared nanofiber can be possible to use for wound healing dressing, patch for skin cancer and other biomedical applications.
456
58532
Cardiolipin-Incorporated Liposomes Carrying Curcumin and Nerve Growth Factor to Rescue Neurons from Apoptosis for Alzheimer’S Disease Treatment
Abstract:
Curcumin (CRM) and nerve growth factor (NGF) were entrapped in liposomes (LIP) with cardiolipin (CL) to downregulate the phosphorylation of mitogen-activated protein kinases for Alzheimer’s disease (AD) management. AD belongs to neurodegenerative disorder with a gradual loss of memory, yielding irreversible dementia. CL-conjugated LIP loaded with CRM (CRM-CL/LIP) and that with NGF (NGF-CL/LIP) were applied to AD models of SK-N-MC cells and Wistar rats with an insult of β-amyloid peptide (Aβ). Lipids comprising 1,2-dipalmitoyl-sn-glycero-3- phosphocholine (Avanti Polar Lipids, Alabaster, AL), 1',3'-bis[1,2- dimyristoyl-sn-glycero-3-phospho]-sn-glycerol (CL; Avanti Polar Lipids), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] (Avanti Polar Lipids), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[carboxy(polyethylene glycol)-2000] (Avanti Polar Lipids) and CRM (Sigma–Aldrich, St. Louis, MO) were dissolved in chloroform (J. T. Baker, Phillipsburg, NJ) and condensed using a rotary evaporator (Panchum, Kaohsiung, Taiwan). Human β-NGF (Alomone Lab, Jerusalem, Israel) was added in the aqueous phase. Wheat germ agglutinin (WGA; Medicago AB, Uppsala, Sweden) was grafted on LIP loaded with CRM for (WGA-CRM-LIP) and CL-conjugated LIP loaded with CRM (WGA-CRM-CL/LIP) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (Sigma–Aldrich) and N-hydroxysuccinimide (Alfa Aesar, Ward Hill, MA). The protein samples of SK-N-MC cells (American Type Tissue Collection, Rockville, MD) were used for sodium dodecyl sulfate (Sigma–Aldrich) polyacrylamide gel (Sigma–Aldrich) electrophoresis. In animal study, the LIP formulations were administered by intravenous injection via a tail vein of male Wistar rats (250–280 g, 8 weeks, BioLasco, Taipei, Taiwan), which were housed in the Animal Laboratory of National Chung Cheng University in accordance with the institutional guidelines and the guidelines of Animal Protection Committee under the Council of Agriculture of the Republic of China. We found that CRM-CL/LIP could inhibit the expressions of phosphorylated p38 (p-p38), p-Jun N-terminal kinase (p-JNK), and p-tau protein at serine 202 (p-Ser202) to retard the neuronal apoptosis. Free CRM and released CRM from CRM-LIP and CRM-CL/LIP were not in a straightforward manner to effectively inhibit the expression of p-p38 and p-JNK in the cytoplasm. In addition, NGF-CL/LIP enhanced the quantities of p-neurotrophic tyrosine kinase receptor type 1 (p-TrkA) and p-extracellular-signal-regulated kinase 5 (p-ERK5), preventing the Aβ-induced degeneration of neurons. The membrane fusion of NGF-LIP activated the ERK5 pathway and the targeting capacity of NGF-CL/LIP enhanced the possibility of released NGF to affect the TrkA level. Moreover, WGA-CRM-LIP improved the permeation of CRM across the blood–brain barrier (BBB) and significantly reduced the Aβ plaque deposition and malondialdehyde level and increased the percentage of normal neurons and cholinergic function in the hippocampus of AD rats. This was mainly because the encapsulated CRM was protected by LIP against a rapid degradation in the blood. Furthermore, WGA on LIP could target N-acetylglucosamine on endothelia and increased the quantity of CRM transported across the BBB. In addition, WGA-CRM-CL/LIP could be effective in suppressing the synthesis of acetylcholinesterase and reduced the decomposition of acetylcholine for better neurotransmission. Based on the in vitro and in vivo evidences, WGA-CRM-CL/LIP can rescue neurons from apoptosis in the brain and can be a promising drug delivery system for clinical AD therapy.
455
59240
Engineering Escherichia coli for Production of Short Chain Fatty Acid by Exploiting Fatty Acid Metabolic Pathway
Abstract:
Worldwide demand of natural and sustainable fuels and chemicals have encouraged researchers to develop microbial platform for synthesis of short chain fatty acids as they are useful precursors to replace petroleum-based fuels and chemicals. In this study, we evaluated the role of fatty acid synthesis and β-oxidation cycle of Escherichia coli to produce butyric acid, a 4-carbon short chain fatty acid, with the help of three thioesterases, i.e., TesAT from Anaerococcus tetradius, TesBF from Bryantella formatexigens and TesBT from Bacteroides thetaiotaomicron. We found that E. coli strain transformed with gene for TesBT and grown in presence of 8 g/L glucose produced maximum butyric acid titer at 1.46 g/L, followed by that of TesBF at 0.85 g/L and TesAT at 0.12 g/L, indicating that these thioesterases were efficiently converting short chain fatty acyl-ACP intermediate of fatty acid synthesis pathway into the corresponding acid. The titer of butyric acid varied significantly depending upon the plasmid copy number and strain genotype. Deletion of genes for fatty acyl-CoA synthetase and acyl-CoA dehydrogenase, which are involved in initiating the fatty acid degradation cycle, and overexpression of FadR, which is a dual transcriptional regulator and exerts negative control over fatty acid degradation pathway, reduced up to 30% of butyric acid titer. This observation suggested that β-oxidation pathway is working synergistically with fatty acid synthesis pathway in production of butyric acid. Moreover, accelerating the fatty acid elongation cycle by overexpressing acetyl-CoA carboxyltransferase (Acc) and 3-hydroxy-acyl-ACP dehydratase (FabZ) or by deleting FabR, the transcription suppressor of elongation, did not improve the butyric acid titer, rather favored the long chain fatty acid production. Finally, a balance between cell growth and butyric acid production was achieved with the use of phosphorous limited growth medium and 14.3 g/L butyric acid, and 17.5 g/L total free fatty acids (FFAs) titer was achieved during fed-batch cultivation. We have engineered an E. coli strain which utilizes the intermediate of both fatty acid synthesis and degradation pathway, i.e. butyryl-ACP and -CoA, to produce butyric acid from glucose. The strategy used in this study resulted in highest reported titers of butyric acid and FFAs in engineered E. coli.
454
58452
Flavonoids and Phenolic Acids from the Aerial Parts of Alyssum alyssoides
Abstract:
Most of Alyssum species of Brassicaceae family have been mainly studied for their contribution in ecology. In this study, A. alyssoides was examined for its chemical substitutes. The methanol extract of its aerial parts was fractionated with liquid-liquid extraction (distribution) with four different solvents of increasing polarity: diethyl ether, ethyl acetate, 1-butanol and water. The diethyl ether and ethyl acetate extracts were further studied for their chemical composition. So far, secondary metabolites which belong to phenolics were isolated by using several chromatographic methods (C.C. and HPLC) and were identified by using spectroscopic methods (UV/Vis, NMR and MS): two phenolic acids (p-hydroxy-benzoic acid and 3-methoxy-4-hydroxy-benzoic acid (vanillic acid)), and five flavonoids, which are derivatives of flavonol: kaempferol 3-O-β-D-glucopyranoside (astragalin), kaempferol 3-O-(6′′-α-L-rhamnopyranosyl)-β-D-glucopyranoside (nicotiflorin), quercetin 3-O-β-D-glucopyranoside (isoquercetin), isorhamnetin-3-O-β-D-glucopyranoside, and isoramnetin 3-O-(6′′-α-L-rhamnopyranosyl)-β-D-glucopyranoside (narcissin).
453
55755
Antimicrobial and Antioxidant Activities of Actinobacteria Isolated from the Pollen of Pinus sylvestris Grown on the Lake Baikal Shore
Abstract:
Isolated ecosystems existing under specific environmental conditions have been shown to be promising sources of new strains of actinobacteria. The taiga forest of Baikal Siberia has not been well studied, and its actinobacterial population remains uncharacterized. The proximity between the huge water mass of Lake Baikal and high mountain ranges influences the structure and diversity of the plant world in Siberia. Here, we report the isolation of eighteen actinobacterial strains from male cones of Pinus sylvestris trees growing on the shore of the ancient Lake Baikal in Siberia. The actinobacterial strains were isolated on solid nutrient MS media and Czapek agar supplemented with cycloheximide and phosphomycin. Identification of actinobacteria was carried out by 16S rRNA gene sequencing and further analysis of the evolutionary history. Four different liquid and solid media (NL19, DNPM, SG and ISP) were tested for metabolite production. The metabolite extracts produced by the isolated strains were tested for antibacterial and antifungal activities. Also, antiradical activity of crude extracts was carried out. Strain Streptomyces sp. IB 2014 I 74-3 that active against Gram-negative bacteria was selected for dereplication analysis with using the high-yield liquid chromatography with mass-spectrometry. Mass detection was performed in both positive and negative modes, with the detection range set to 160–2500 m/z. Data were collected and analyzed using Bruker Compass Data Analysis software, version 4.1. Dereplication was performed using the Dictionary of Natural Products (DNP) database version 6.1 with the following search parameters: accurate molecular mass, absorption spectra and source of compound isolation. Thus, in addition to more common representative strains of Streptomyces, several species belonging to the genera Rhodococcus, Amycolatopsis, and Micromonospora were isolated. Several of the selected strains were deposited in the Russian Collection of Agricultural Microorganisms (RCAM), St. Petersburg, Russia. All isolated strains exhibited antibacterial and antifungal activities. We identified several strains that inhibited the growth of the pathogen Candida albicans but did not hinder the growth of Saccharomyces cerevisiae. Several isolates were active against Gram-positive and Gram-negative bacteria. Moreover, extracts of several strains demonstrated high antioxidant activity. The high proportion of biologically active strains producing antibacterial and specific antifungal compounds may reflect their role in protecting pollen against phytopathogens. Dereplication of the secondary metabolites of the strain Streptomyces sp. IB 2014 I 74-3 was resulted in the fact that a total of 59 major compounds were detected in the culture liquid extract of strain cultivated in ISP medium. Eight compounds were preliminarily identified based on characteristics described in the Dictionary of Natural Products database, using the search parameters Streptomyces sp. IB 2014 I 74-3 was found to produce saframycin A, Y3 and S; 2-amino-3-oxo-3H-phenoxazine-1,8-dicarboxylic acid; galtamycinone; platencin A4-13R and A4-4S; ganefromycin d1; the antibiotic SS 8201B; and streptothricin D, 40-decarbamoyl, 60-carbamoyl. Moreover, forty-nine of the 59 compounds detected in the extract examined in the present study did not result in any positive hits when searching within the DNP database and could not be identified based on available mass-spec data. Thus, these compounds might represent new findings.