Search results for: enzyme linked immunosorbent assay

Authors: Jyh-Cherng Gu, Ming-Ta Yang, Dai-Ling Tsai

Abstract:

Both sparse lashing and dense lashing are applied to secure overhead XLPE (cross-linked polyethylene) covered power lines on ceramic insulators or HDPE polymer insulators. The distribution of electric field in and among the lashing wires, the XLPE power lines and insulators in normal clean condition and when conducting materials such as salt, metal particles, dust, smoke or acidic smog are present is studied in this paper. The ANSYS Maxwell commercial software is used in this study for electric field analysis. Although the simulation analysis is performed assuming ideal conditions due to the constraints of the simulation software, the result may not be the same as in real situation but still be of sufficient practical values.

Keywords: electric field intensity, insulator, XLPE covered aerial line, empty

Procedia PDF Downloads 261

Authors: Natarajan Ramasamy, Gurulingamurthy Haralur, Ramesh Nivarthu, Nikhil Kumar Singha

Abstract:

Elastomers are polymeric materials with varied backbone architectures ranging from linear to dendrimeric structures and wide varieties of monomeric repeat units. These elastomers show strongly viscous and weakly elastic when it is not cross-linked. But when crosslinked, based on the extent the properties of these elastomers can range from highly flexible to highly stiff nature. Lightly cross-linked systems are well studied and reported. Understanding the nature of highly cross-linked rubber based upon chemical structure and architecture is critical for varieties of applications. One of the critical parameters is cross-link density. In the current work, we have studied the highly cross-linked state of linear, lightly branched to star-shaped branched elastomers and determined the cross-linked density by using different models. Change in hardness, shift in Tg, change in modulus and swelling behavior were measured experimentally as a function of the extent of curing. These properties were analyzed using varied models to determine cross-link density. We used hardness measurements to examine cure time. Hardness to the extent of curing relationship is determined. It is well known that micromechanical transitions like Tg and storage modulus are related to the extent of crosslinking. The Tg of the elastomer in different crosslinked state was determined by DMA, and based on plateau modulus the crosslink density is estimated by using Nielsen’s model. Usually for lightly crosslinked systems, based on equilibrium swelling ratio in solvent the cross link density is estimated by using Flory–Rhener model. When it comes to highly crosslinked system, Flory-Rhener model is not valid because of smaller chain length. So models based on the assumption of polymer as a Non-Gaussian chain like 1) Helmis–Heinrich–Straube (HHS) model, 2) Gloria M.gusler and Yoram Cohen Model, 3) Barbara D. Barr-Howell and Nikolaos A. Peppas model is used for estimating crosslink density. In this work, correction factors are determined to the existing models and based upon it structure-property relationship of highly crosslinked elastomers was studied.

Keywords: dynamic mechanical analysis, glass transition temperature, parts per hundred grams of rubber, crosslink density, number of networks per unit volume of elastomer

Procedia PDF Downloads 160

Authors: Amarjyoti Das Mahapatra, Umesh Kumar, Bhaskar Datta

Abstract:

A pseudo peptide can mimic the biological or structural properties of natural peptides. They have become an increasing attention in medicinal chemistry because of their interesting advantages like more bioavailability and less biodegradation than compare to the physiologically active native peptides which increase their therapeutic applications. Many biologically active compounds contain urea as functional groups, and they have improved pharmacokinetic properties because of their bioavailability and metabolic stability. Recently we have reported a single-step synthesis of sulfonyl urea and sulfonyltriuret from sulfonyl chloride and sodium cyanate. But the yield of sulfonyltriuret was less around 40-60% because of the formation of other products like sulfonamide and sulfonylureas. In the present work, we mainly focused on the selective synthesis of sulfonyltriuret using tetrabutylammonium cyanate and sulfonyl chloride. More precisely, we are interested in the controlled synthesis of oligomeric urea mainly sulfonyltriuret as a new class of pseudo peptide and their application as protease modulators. The distinctive architecture of these molecules in the form of their pseudo-peptide backbone offers promise as a potential pharmacophore. The synthesized molecules have been screened on trypsin enzyme, and we observed that these molecules are the efficient modulator of trypsin enzyme.

Keywords: pseudo peptide, pharmacophore, sulfonyltriuret, trypsin

Procedia PDF Downloads 160

Authors: Fadime Eroglu

Abstract:

Acanthamoeba genus belongs to kingdom protozoa, and it is known as free-living amoebae. Acanthamoeba genus has been isolated from human bodies, swimming pools, bottled mineral water, contact lens solutions, dust, and soil. The members of the genus Acanthamoeba causes Acanthamoeba Keratitis which is a painful sight-threatening disease of the eyes. In recent years, the prevalence of Acanthamoeba keratitis has been high rate reported. The eight different Acanthamoeba species are known to be effective in Acanthamoeba keratitis. These species are Acanthamoeba castellanii, Acanthamoeba polyphaga, Acanthamoeba griffini, Acanthamoeba hatchetti, Acanthamoeba culbertsoni and Acanhtamoeba rhysodes. The conventional diagnosis of Acanthamoeba Keratitis has relied on cytological preparations and growth of Acanthamoeba in culture. However molecular methods such as real-time PCR has been found to be more sensitive. The real-time PCR has now emerged as an effective method for more rapid testing for the diagnosis of infectious disease in decade. Therefore, a real-time PCR assay for the detection of Acanthamoeba keratitis and Acanthamoeba species have been developed in this study. The 18S rRNA sequences from Acanthamoeba species were obtained from National Center for Biotechnology Information and sequences were aligned with MEGA 6 programme. Primers and probe were designed using Custom Primers-OligoPerfectTMDesigner (ThermoFisherScientific, Waltham, MA, USA). They were also assayed for hairpin formation and degree of primer-dimer formation with Multiple Primer Analyzer ( ThermoFisherScientific, Watham, MA, USA). The eight different ATCC Acanthamoeba species were obtained, and DNA was extracted using the Qiagen Mini DNA extraction kit (Qiagen, Hilden, Germany). The DNA of Acanthamoeba species were analyzed using newly designed primer and probe set in real-time PCR assay. The early definitive laboratory diagnosis of Acanthamoeba Keratitis and the rapid initiation of suitable therapy is necessary for clinical prognosis. The results of the study have been showed that new primer and probes could be used for detection and distinguish for Acanthamoeba species. These new developing methods are helpful for diagnosis of Acanthamoeba Keratitis.

Keywords: Acathamoeba Keratitis, Acanthamoeba species, fast diagnosis, Real-Time PCR

Procedia PDF Downloads 115

Authors: Busra Gundede, Ozal Mutlu, Nagihan Gulsoy

Abstract:

Background: Over the years, material research has led to the development of numerous metals and alloys for using in biomedical applications. One of the major tasks of biomaterial research is the functionalization of the material surface to improve the biocompatibility according to a specific application. 316L and 316L alloys are excellent for various bio-applications. This research was investigated the effect of titanium (Ti), niobium (Nb), and zirconium (Zr) additives on injection molded austenitic grade 316L stainless steels in vitro biocompatibility. For this purpose, cytotoxic tests were performed to evaluate the potential biocompatibility of the specimens. Materials and Methods: 3T3 fibroblast were cultivated in DMEM supplemented with 10% fetal bovine serum and %1 penicillin-streptomycin at 37°C with 5% CO2 and 95%humidity. Trypsin/EDTA solution was used to remove cells from the culture flask. Cells were reseeded at a density of 1×105cell in 25T flasks. The medium change took place every 3 days. The trypan blue assay was used to determine cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the cell counter machine counted the number of blue staining cells and the number of total cells. Cell viability should be at least 95% for healthy log-phase cultures. MTT assay was assessed for 96-hours. Cells were cultivated in 6-well flask within 5 ml DMEM and incubated as same conditions. 0,5mg/ml MTT was added for 4-hours and then acid-isoprohanol was added for solubilize to formazan crystals. Cell morphology after 96h was investigated by SEM. The medium was removed, samples were washed with 0.15 M PBS buffer and fixed for 12h at 4- 8°C with %2,5 gluteraldehyte. Samples were treated with 1% osmium tetroxide. Samples were then dehydrated and dried, mounted on appropriate stubs with colloidal silver and sputter-coated with gold. Images were collected using a scanning electron microscope. ROS assay is a cell viability test for in vitro studies. Cells were grown for 96h, ROS solution added on cells in 6 well plate flask and incubated for 1h. Fluorescence signal indicates ROS generation by cells. Results: Trypan Blue exclusion assay results were 96%, 92%, 95%, 90%, 91% for negative control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Results were found nearly similar to each other when compared with control group. Cell viability from MTT analysis was found to be 100%, 108%, 103%, 107%, and 105% for the control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Fluorescence microscopy analysis indicated that all test groups were same as the control group in ROS assay. SEM images demonstrated that the attachment of 3T3 cells on biomaterials. Conclusion: We, therefore, concluded that Ti, Nb and Zr additives improved physical properties of 316L stainless. In our in vitro experiments showed that these new additives did not modify the cytocompatibility of stainless steel and these additives on 316L might be useful for biomedical applications.

Keywords: 316L stainles steel, biocompatibility, cell culture, Ti, Nb, Zr

Procedia PDF Downloads 511

Authors: Lorena C. Cintra, Izadora M. De Oliveira, Amanda G. Fernandes, Francieli Colussi, Rosália S. A. Jesuíno, Fabrícia P. Faria, Cirano J. Ulhoa

Abstract:

Xylan is the main component of hemicellulose and for its complete degradation is required cooperative action of a system consisting of several enzymes including endo-xylanases (XYN), β-xylosidases (XYL) and α-L-arabinofuranosidases (ABF). The recombinant hemicellulolytic enzymes an endoxylanase (HXYN2), β-xylosidase (HXYLA), and an α-L-arabinofuranosidase (ABF3) were used in hydrolysis tests. These three enzymes are produced by filamentous fungi and were expressed heterologously and produced in Pichia pastoris previously. The aim of this work was to evaluate the effect of recombinant hemicellulolytic enzymes on the enzymatic hydrolysis of sugarcane bagasse (SCB). The interaction between the three recombinant enzymes during SCB pre-treated by steam explosion hydrolysis was performed with different concentrations of HXYN2, HXYLA and ABF3 in different ratios in according to a central composite rotational design (CCRD) 23, including six axial points and six central points, totaling 20 assays. The influence of the factors was assessed by analyzing the main effects and interaction between the factors, calculated using Statistica 8.0 software (StatSoft Inc. Tulsa, OK, USA). The Pareto chart was constructed with this software and showed the values of the Student’s t test for each recombinant enzyme. It was considered as response variable the quantification of reducing sugars by DNS (mg/mL). The Pareto chart showed that the recombinant enzyme ABF3 exerted more significant effect during SCB hydrolysis, with higher concentrations and with the lowest concentration of this enzyme. It was performed analysis of variance according to Fisher method (ANOVA). In ANOVA for the release of reducing sugars (mg/ml) as the variable response, the concentration of ABF3 showed significance during hydrolysis SCB. The result obtained by ANOVA, is in accordance with those presented in the analysis method based on the statistical Student's t (Pareto chart). The degradation of the central chain of xylan by HXYN2 and HXYLA was more strongly influenced by ABF3 action. A model was obtained, and it describes the performance of the interaction of all three enzymes for the release of reducing sugars, and can be used to better explain the results of the statistical analysis. The formulation capable of releasing the higher levels of reducing sugars had the following concentrations: HXYN2 with 600 U/g of substrate, HXYLA with 11.5 U.g-1 and ABF3 with 0.32 U.g-1. In conclusion, the recombinant enzyme that has a more significant effect during SCB hydrolysis was ABF3. It is noteworthy that the xylan present in the SCB is arabinoglucoronoxylan, due to this fact debranching enzymes are important to allow access of enzymes that act on the central chain.

Keywords: experimental design, hydrolysis, recombinant enzymes, sugar cane bagasse

Procedia PDF Downloads 225

Authors: Daniela N. Correa-Llantén, Sebastián A. Muñoz-Ibacache, Mathilde Maire, Jenny M. Blamey

Abstract:

The biosynthesis of nanoparticles by microorganisms, on the contrary to chemical synthesis, is an environmentally-friendly process which has low energy requirements. In this investigation, we used the microorganism Geobacillus wiegelii, strain GWE1, an aerobic thermophile belonging to genus Geobacillus, isolated from a drying oven. This microorganism has the ability to reduce selenite evidenced by the change of color from colorless to red in the culture. Elemental analysis and composition of the particles were verified using transmission electron microscopy and energy-dispersive X-ray analysis. The nanoparticles have a defined spherical shape and a selenium elemental state. Previous experiments showed that the presence of the whole microorganism for the reduction of selenite was not necessary. The results strongly suggested that an intracellular NADPH/NADH-dependent reductase mediates selenium nanoparticles synthesis under aerobic conditions. The enzyme was purified and identified by mass spectroscopy MALDI-TOF TOF technique. The enzyme is a 1-pyrroline-5-carboxylate dehydrogenase. Histograms of nanoparticles sizes were obtained. Size distribution ranged from 40-160 nm, where 70% of nanoparticles have less than 100 nm in size. Spectroscopic analysis showed that the nanoparticles are composed of elemental selenium. To analyse the effect of pH in size and morphology of nanoparticles, the synthesis of them was carried out at different pHs (4.0, 5.0, 6.0, 7.0, 8.0). For thermostability studies samples were incubated at different temperatures (60, 80 and 100 ºC) for 1 h and 3 h. The size of all nanoparticles was less than 100 nm at pH 4.0; over 50% of nanoparticles have less than 100 nm at pH 5.0; at pH 6.0 and 8.0 over 90% of nanoparticles have less than 100 nm in size. At neutral pH (7.0) nanoparticles reach a size around 120 nm and only 20% of them were less than 100 nm. When looking at temperature effect, nanoparticles did not show a significant difference in size when they were incubated between 0 and 3 h at 60 ºC. Meanwhile at 80 °C the nanoparticles suspension lost its homogeneity. A change in size was observed from 0 h of incubation at 80ºC, observing a size range between 40-160 nm, with 20% of them over 100 nm. Meanwhile after 3 h of incubation at size range changed to 60-180 nm with 50% of them over 100 nm. At 100 °C the nanoparticles aggregate forming nanorod structures. In conclusion, these results indicate that is possible to modulate size and shape of biologically synthesized nanoparticles by modulating pH and temperature.

Keywords: genus Geobacillus, NADPH/NADH-dependent reductase, selenium nanoparticles, biosynthesis

Procedia PDF Downloads 309

Authors: Gajanan M. Sonwane

Abstract:

Nowadays, the entire pharmaceutical industry is facing the challenge of increasing efficiency and innovation. The major hurdles are the growing cost of research and development and a concurrent stagnating number of new chemical entities (NCEs). Hence, the challenge is to select the most druggable targets and to search the equivalent drug-like compounds, which also possess specific pharmacokinetic and toxicological properties that allow them to be developed as drugs. The present research work includes the studies of developing new anticancer heterocycles by using molecular modeling techniques. The heterocycles synthesized through such methodology are much effective as various physicochemical parameters have been already studied and the structure has been optimized for its best fit in the receptor. Hence, on the basis of the literature survey and considering the need to develop newer anticancer agents, new phenazinamine derivatives were designed by subjecting the nucleus to molecular modeling, viz., GQSAR analysis and docking studies. Simultaneously, these designed derivatives were subjected to in silico prediction of biological activity through PASS studies and then in silico toxicity risk assessment studies. In PASS studies, it was found that all the derivatives exhibited a good spectrum of biological activities confirming its anticancer potential. The toxicity risk assessment studies revealed that all the derivatives obey Lipinski’s rule. Amongst these series, compounds 4c, 5b and 6c were found to possess logP and drug-likeness values comparable with the standard Imatinib (used for anticancer activity studies) and also with the standard drug methotrexate (used for antimitotic activity studies). One of the most notable mutations is the threonine to isoleucine mutation at codon 315 (T315I), which is known to be resistant to all currently available TKI. Enzyme assay planned for confirmation of target selective activity.

Keywords: drug design, tyrosine kinases, anticancer, Phenazinamine

Procedia PDF Downloads 110

Authors: S. Beekrum, B. Odhav, R. Lalloo, E. O. Amonsou

Abstract:

Marine microalgae are rich sources of the novel and biologically active metabolites; therefore, they may be used in the food industry as natural food ingredients and functional foods. They have several biological applications related with health benefits, among others. In the past decades, food scientists have been searching for natural alternatives to replace synthetic antioxidants. The use of synthetic antioxidants has decreased due to their suspected activity as promoters of carcinogenesis, as well as consumer rejection of synthetic food additives. The aim of the study focused on screening of phytochemicals from Cymbella biomass extracts, and to examine the in vitro antioxidant and antimicrobial potential. Cymbella biomass was obtained from CSIR (South Africa), and four different solvents namely methanol, acetone, n-hexane and water were used for extraction. To take into account different antioxidant mechanisms, seven different antioxidant assays were carried out. These include free radical scavenging (DPPH assay), Trolox equivalent antioxidant capacity (TEAC assay), radical cation (ABTS assay), superoxide anion radical scavenging, reducing power, determination of total phenolic compounds and determination of total flavonoid content. The total content of phenol and flavonoid in extracts were determined as gallic acid equivalent, and as rutin equivalent, respectively. The in vitro antimicrobial effect of extracts were tested against some pathogens (Staphylococcus aureus, Listeria monocytogenes, Bacillus subtilis, Salmonella enteritidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans), using the disc diffusion assay. Qualitative analyses of phytochemicals were conducted by chemical tests to screen for the presence of tannins, flavonoids, terpenoids, phenols, steroids, saponins, glycosides and alkaloids. The present investigation revealed that all extracts showed relatively strong antibacterial activity against most of the tested bacteria. The methanolic extract of the biomass contained a significantly high phenolic content of 111.46 mg GAE/g, and the hexane extract contained 65.279 mg GAE/g. Results of the DPPH assay showed that the biomass contained strong antioxidant capacity, 79% in the methanolic extract and 85% in the hexane extract. Extracts have displayed effective reducing power and superoxide anion radical scavenging. Results of this study have highlighted potential antioxidant activity in the methanol and hexane extracts. The obtained results of the phytochemical screening showed the presence of terpenoids, flavonoids, phenols and saponins. The use of Cymbella as a natural antioxidant source and a potential source of antibacterial compounds and phytochemicals in the food industry appears promising and should be investigated further.

Keywords: antioxidants, antimicrobial, Cymbella, microalgae, phytochemicals

Procedia PDF Downloads 448

Authors: Aouissi Nora, Meksem Leila

Abstract:

This work focused on the study of air pollution generated by the transport sector in the region of Annaba. Our study is based on two parts: the first one concerns an epidemiological investigation in the area of Annaba situated in the east Algerian coast, which deals with the development of the fleet and its impact on public health. To get a more precise idea of the impact of road traffic on public health, we consulted the computing center office of the National Social Insurance Fund. The information we were given by this office refers to the number of reported asthma and heart disease after medical examination during the period 2006-2010. The second part was devoted to the study of the toxicity of exhaust gases on some physical and biochemical parameters of durum wheat (Triticum durum Desf.). After germination and three-leaf stage, the pots are placed in a box of volume (0,096 m3) having an input which is linked directly to the exhaust pipe of a truck, and an outlet to prevent asphyxiation plant. The experience deals with 30 pots: 10 pots are exposed for 5 minutes to exhaust smoke; the other 10 are exposed for 15 minutes, and the remaining 10 for 30 minutes. The epidemiological study shows that the levels of pollutants emitted by the fleet are responsible for the increase of people respiratory and cardiovascular diseases. As for biochemical analyses of vegetation, they clearly show the toxicity of pollutants emitted by the exhaust gases, with an increase in total protein, proline and stimulation of detoxification enzyme (catalase).

Keywords: air pollution, toxicity, epidemiology, biochemistry

Procedia PDF Downloads 329

Authors: Prince Kumar, Shamseer Kulangara Kandi, Diwan S. Rawat, Kasturi Mukhopadhyay

Abstract:

Staphylococcus aureus is the most pathogenic of all staphylococci, a major cause of nosocomial infections, and known for acquiring resistance towards various commonly used antibiotics. Due to the widespread use of synthetic drugs, clinicians are now facing a serious threat in healthcare. The increasing resistance in staphylococci has created a need for alternatives to these synthetic drugs. One of the alternatives is a natural plant-based medicine for both disease prevention as well as the treatment of chronic diseases. Among such natural compounds, curcumin is one of the most studied molecules and has been an integral part of traditional medicines and Ayurveda from ancient times. It is a natural polyphenolic compound with diverse pharmacological effects, including anti-inflammatory, antioxidant, anti-cancerous and antibacterial activities. In spite of its efficacy and potential, curcumin has not been approved as a therapeutic agent yet, because of its low solubility, low bioavailability, and rapid metabolism in vivo. The presence of central β-diketone moiety in curcumin is responsible for its rapid metabolism. To overcome this, in the present study, curcuminoids were designed by modifying the central β-diketone moiety of curcumin into mono carbonyl moiety and their antibacterial potency against S. aureus ATCC 29213 was determined. Further, the mode of action and hemolytic activity of the most potent curcuminoids were studied. Minimum inhibitory concentration (MIC) and in vitro killing kinetics were used to study the antibacterial activity of the designed curcuminoids. For hemolytic assay, mouse Red blood cells were incubated with curcuminoids and hemoglobin release was measured spectrophotometrically. The mode of action of curcuminoids was analysed by membrane depolarization assay using membrane potential sensitive dye 3,3’-dipropylthiacarbocyanine iodide (DiSC3(5)) through spectrofluorimetry and membrane permeabilization assay using calcein-AM through flow cytometry. Antibacterial screening of the designed library (61 curcuminoids) revealed excellent in vitro potency of six compounds against S. aureus (MIC 8 to 32 µg/ml). Moreover, these six compounds were found to be non-hemolytic up to 225 µg/ml that is much higher than their corresponding MIC values. The in vitro killing kinetics data showed five of these lead compounds to be bactericidal causing >3 log reduction in the viable cell count within 4 hrs at 5 × MIC while the sixth compound was found to be bacteriostatic. Depolarization assay revealed that all the six curcuminoids caused depolarization in their corresponding MIC range. Further, the membrane permeabilization assay showed that all the six curcuminoids caused permeabilization at 5 × MIC in 2 hrs. This membrane depolarization and permeabilization caused by curcuminoids found to be in correlation with their corresponding killing efficacy. Both these assays point out that membrane perturbations might be a primary mode of action for these curcuminoids. Overall, the present study leads us six water soluble, non-hemolytic, membrane-active curcuminoids and provided an impetus for further research on therapeutic use of these lead curcuminoids against S. aureus.

Keywords: antibacterial, curcumin, minimum inhibitory concentration , Staphylococcus aureus

Procedia PDF Downloads 166

Authors: Tiantian Min, Chuanxiang Cheng, Jin Yue

Abstract:

The growth and reproduction of microorganisms in food packaging can cause food decay and foodborne diseases, which pose a serious threat to the health of consumers and even cause serious economic losses. Active food packaging containing antibacterial bioactive compounds is a promising strategy for extending the shelf life of products and maintaining the food quality, as well as reducing the food waste. However, most active packaging can only act as slow-release effect for antimicrobials, which causes the release rate of antimicrobials not match the growth rate of microorganisms. Stimuli-responsive active packaging materials based on biopolymeric substrates and bioactive substances that respond to some biological and non-biological trigger factors provide more opportunities for fresh food preservation. The biological stimuli factors such as relative humidity, pH and enzyme existed in the exudate secreted by microorganisms have been expected to design food packaging materials. These stimuli-responsive materials achieved accurate release or delivery of bioactive substances at specific time and appropriate dose. Recently, metal-organic-frameworks (MOFs) nanoparticles become attractive carriers to enhance the efficiency of bioactive compounds or drugs. Cellulose nanofibrils have been widely applied for film substrates due to their biodegradability and biocompatibility. The abundant hydroxyl groups in cellulose can be oxidized to carboxyl groups by TEMPO, making it easier to anchoring MOFs and to be further modification. In this study, a pH and enzyme dual-responsive CAR@ZIF-8/TOCNF/PE film was fabricated by in-situ growth of ZIF-8 nanoparticles onto TEMPO-oxidized cellulose (TOCNF) film and further coated with pectin (PE) for stabilization and controlled release of carvacrol (CAR). The enzyme triggered release of CAR was achieved owing to the degradation of pectin by pectinase secreted by microorganisms. Similarly, the pH-responsive release of CAR was attributed to the unique skeleton degradation of ZIF-8, further accelerating the release of CAR from the topological structure of ZIF-8. The composite film performed excellent crystallinity and adsorb ability confirmed by X-ray diffraction and BET analysis, and the inhibition efficiency against Escherichia coli, Staphylococcus aureus and Aspergillus niger reached more than 99%. The composite film was capable of releasing CAR when exposure to dose-dependent enzyme (0.1, 0.2, and 0.3 mg/mL) and acidic condition (pH = 5). When inoculated 10 μL of Aspergillus niger spore suspension on the equatorial position of mango and raspberries, this composite film acted as packaging pads effectively inhibited the mycelial growth and prolonged the shelf life of mango and raspberries to 7 days. Such MOF-TOCNF based film provided a targeted, controlled and sustained release of bioactive compounds for long-term antibacterial activity and preservation effect, which can also avoid the cross-contamination of fruits.

Keywords: active food packaging, controlled release, fruit preservation, in-situ growth, stimuli-responsive

Procedia PDF Downloads 56

Authors: Nadine Wiesmann, Melanie Viel, Christoph Buhr, Rachel Tanner, Wolfgang Tremel, Juergen Brieger

Abstract:

Recidivation of tumors and the development of resistances against the classical anti-tumor approaches represent a major challenge we face when treating cancer. In order to master this challenge, we are in desperate need of new treatment options beyond the beaten tracks. Zinc oxide nanoparticles (ZnO NPs) represent such an innovative approach. Zinc oxide is characterized by a high level of biocompatibility, concurrently ZnO NPs are able to exert anti-tumor effects. By concentration of the nanoparticles at the tumor site, tumor cells can specifically be exposed to the nanoparticles while low zinc concentrations at off-target sites are tolerated well and can be excreted easily. We evaluated the toxicity of ZnO NPs in vitro with the help of immortalized tumor cell lines and primary cells stemming from healthy tissue. Additionally, the Chorioallantoic Membrane Assay (CAM Assay) was employed to gain insights into the in vivo behavior of the nanoparticles. We could show that ZnO NPs interact with tumor cells as nanoparticulate matter. Furthermore, the extensive release of zinc ions from the nanoparticles nearby and within the tumor cells results in overload with zinc. Beyond that, ZnO NPs were found to further the generation of reactive oxygen species (ROS). We were able to show that tumor cells were more prone to the toxic effects of ZnO NPs at intermediate concentrations compared to fibroblasts. With the help of ZnO NPs covered by a silica shell in which FITC dye was incorporated, we were able to track ZnO NPs within tumor cells as well as within a whole organism in the CAM assay after injection into the bloodstream. Depending on the applied concentrations, selective tumor cell killing seems feasible. Furthermore, the combinational treatment of tumor cells with radiotherapy and ZnO NPs shows promising results. Still, further investigations are needed to gain a better understanding of the interaction between ZnO NPs and the human body to be able to pave the way for their application as an innovative anti-tumor agent in the clinics.

Keywords: metal oxide nanoparticles, nanomedicine, overcome resistances against classical treatment options, zinc oxide nanoparticles

Procedia PDF Downloads 123

Authors: Asma Meliani, S. Lakehal, F. Z. Benrebiha, C. Chaouia

Abstract:

There is an increasing interest in phytochemicals as new source of natural antioxidant and antimicrobial agents. Plants essential oils have come more into the focus of phytomedicine. Many researchers have reported various biological and/or pharmacological properties of Artemisia herba alba Asso essential oil. The present study describes antimicrobial and antioxidant properties of Artemisia herba alba Asso essential oil. Artemisia herba alba Asso essential oil obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (M’sila) was analyzed by GC-MS. The essential oil yield of the study was 0.7%. The major components were found to be camphor, chrysanthenone et 1,8-cineole. The antimicrobial activity of the essential oil was tested against four bacteria (Gram-negative and Gram-positive) and three fungi using the diffusion method and by determining the inhibition zone. The oil was found to have significant antibacterial activity. In addition, antioxidant activity was determined by 1, 1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and β-carotene bleaching test, and high activity was found for Artemisia herba-alba oil.

Keywords: Artemisia herba-alba, essential oil, antibacterial activity, antioxidant activity

Procedia PDF Downloads 323

Authors: Afshin Farahbakhsh, Hoda Khodadadi

Abstract:

In this paper, we compare five methods of biological fuel cell fabrication by combining a Shewanella oneidensis microbial anode and a laccase-modified air-breathing cathode. As a result of biofuel cell laccase with graphite nanofibers, carbon surface (PAMAN) on the pt/hpg electrode, graphite sheets MWCNT and with (PG) and (MWCNT) showed, respectively. Describes methods for creating controllable and reproducible bio-anodes and demonstrates the versatility of hybrid biological fuel cells. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. The laccase-based biocathodes prepared either with the crude extract or with the purified enzyme can provide electrochemically active and stable biomaterials. When the device was fed with transdermal extracts, containing only 30μM of glucose, the average peak power was proportionally lower (0.004mW). The result of biofuel cell with graphite nanofibers showed the enzymatic fuel cell reaches 0.5 V at open circuit voltage with both, ethanol and methanol and the maximum current density observed for E2electrode was 228.94mAcm.

Keywords: enzymatic electrode, fuel cell, immobilization, laccase

Procedia PDF Downloads 255

Authors: Asma Meliani, S. Lakehal, F. Z. Benrebiha, C. Chaouia

Abstract:

There is an increasing interest in phytochemicals as new source of natural antioxidant and antimicrobial agents. Plants essential oils have come more into the focus of phytomedicine. Many researchers have reported various biological and/or pharmacological properties of Artemisia herba alba Asso essential oil. The present study describes antimicrobial and antioxidant properties of Artemisia herba alba Asso essential oil. Artemisia herba alba Asso essential oil obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (M’sila) was analyzed by GC-MS. The essential oil yield of the study was 0.7 %. The major components were found to be camphor, chrysanthenone et 1,8-cineole. The antimicrobial activity of the essential oil was tested against four bacteria (Gram-negative and Gram-positive) and one fungi using the diffusion method and by determining the inhibition zone. The oil was found to have significant antibacterial activity. In addition, antioxidant activity was determined by 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and β-carotene bleaching test, and high activity was found for Artemisia herba-alba oil.

Keywords: Artemisia herba-alba, essential oil, antibacterial activity, antioxidant activity

Procedia PDF Downloads 459

Authors: Fadia Gafri, Kathryn Mckintosh, Louise Young, Alan Harvey, Simon Mackay, Andrew Paul, Robin Plevin

Abstract:

Nuclear factor-kappa B (NF-κB) is a ubiquitous transcription factor found originally to play a key role in regulating inflammation. However considerable evidence links this pathway to the suppression of apoptosis, cellular transformation, proliferation and invasion (Aggarwal et al., 2006). Moreover, recent studies have also linked inflammation to cancer progression making NF-κB overall a promising therapeutic target for drug discovery (Dobrovolskaia & Kozlov, 2005). In this study we examined the effect of the natural product canthin-6-one (SU182) as part of a CRUK small molecule drug discovery programme for effects upon the NF-κB pathway. Initial studies demonstrated that SU182 was found to have good potency against the inhibitory kappa B kinases (IKKs) at 30M in vitro. However, at concentrations up to 30M, SU182 had no effect upon TNFα stimulated loss in cellular IκBα or p65 phosphorylation in the keratinocyte cell line NCTC2544. Nevertheless, 30M SU182 reduced TNF-α / PMA-induced NF-κB-linked luciferase reporter activity to (22.9 ± 5%) and (34.6± 3 %, P<0.001) respectively, suggesting an action downstream of IKK signalling. Indeed, SU182 neither decreased NF-κB-DNA binding as assayed by EMSA nor prevented the translocation of p65 (NF-κB) to the nucleus assessed by immunofluorescence and subcellular fractionation. In addition to the inhibition of transcriptional activity of TNFα-induced NF-κB reporter activity SU182 significantly reduced PMA-induced AP-1-linked luciferase reporter activity to about (48± 9% at 30M, P<0.001) . This mode of inhibition was not sufficient to prevent the activation of NF-κB dependent induction of other proteins such as COX-2 and iNOS, or activated MAP kinases (p38, JNK and ERK1/2) in LPS stimulated RAW 264.7 macrophages. Taken together these data indicate the potential for SU182 to interfere with the transcription factors NF-κB and AP-1 at transcriptional level. However, no potential anti-inflammatory effect was indicated, further investigation for other NF-κB dependent proteins linked to survival are also required to identify the exact mechanism of action.

Keywords: Canthin-6-one, NF-κB, AP-1, phosphorylation, Nuclear translocation, DNA-binding activity, inflammatory proteins.

Procedia PDF Downloads 451

Authors: J. Janicki, M. Rom, C. Slusarczyk, J. Fabia, M. Siika-aho, K. Marjamaa, K. Kruus, K. Langfelder, C. Steel, M. Paloheimo, T. Puranen, S. Mäkinen, D. Wawro

Abstract:

The cellulose is one of the most abundant polymers in the world, however, its application in the high-end value products such as films or fibres, it triggered by the cellulose properties. The noticeable presence of hydrogen bonding reflected with partially crystalline structure makes the cellulose insoluble in common solvents and not meltable. The existing technologies, such as viscose process, suffer from environmental and economical problems, because of the risk of harmful chemicals liberation during the spinning process. The enzymatic modification of cellulose with endoglucanase makes it directly alkali soluble in NaOH solution, giving the opportunities for film and fibers formation. As the effect of enzymatic treatment, there are observed changes in crystalline structure and accompanying changes of the affinity of cellulose to water, demonstrated by water retention value. The objective of the project ELMO - Novel carbohydrate modifying enzymes for fibre modification is is to develop new enzyme products for modification of dissolving grade pulps. The aim is to increase the reactivity of dissolving grade pulps and remove residual hemicellulose. The scientific aim of this paper is to present the effect of enzymatic treatment on the crystallinity and affinity to water of cellulose pulps modified with enzymes.

Keywords: cellulose, crystallinity, WAXS, enzyme

Procedia PDF Downloads 230

Authors: Hoo Cheol Lee, Dae Seung Kim, Sang Myung Jung, Gwang Heum Yoon, Hwa Sung Shin

Abstract:

Loss of bone tissue can occur due to a bone tissue disease and aging or fracture. Renewable formation of bone is mainly made by its differentiation and metabolism. For this reason, osteoblasts have been studied for regeneration of bone tissue. So, tissue engineering has attracted attention as a recovery means. In tissue engineering, a particularly important factor is a scaffold that supports cell growth. For osteoblast scaffold, we used the cellulose nanowhisker (CNW) extracted from marine organism. CNW is one of an abundant material obtained from a number of plants and animals. CNW is polymer consisting of monomer cellulose and this composition offers biodegradability and biocompatibility to CNW. Mechanical strength of CNW is superior to the existing natural polymers. In addition, substances of marine origin have a low risk of secondary infection by bacteria and pathogen in contrast with those of land-derived. For evaluating its suitability as an osteoblast scaffold, we fabricate CNW film for osteoblast culture and performed the MTT assay and ALP assay to confirm its cytotoxicity and effect on differentiation. Taking together these results, we assessed CNW is a potential candidate of a material for bone tissue regeneration.

Keywords: bone regeneration, cellulose nanowhisker, marine derived material, osteoblast

Procedia PDF Downloads 340

Authors: A. T. Ramachandra Naik, H. Shivananda Murthy, H. n. Anjanayappa

Abstract:

The freshwater prawn, Macrobrachium rosenbergii is a more popular crustacean cultured widely in monoculture system in India. It has got high nutritional value in the human diet. Hence, understanding its enzymatic and body composition is important in order to judge its flesh quality. Fish oil specially derived from Indian oil sardine is a good source of highly unsaturated fatty acid and lipid source in fish/prawn diet. A 35% crude protein diet with graded levels of Sardine oil as a source of fat was incorporated at four levels viz, 2.07, 4.07, 6.07 and 8.07% maintaining a total lipid level of feed at 8.11, 10.24, 12.28 and 14.33% respectively. Diet without sardine oil (6.05% total lipid) was served as basal treatment. The giant freshwater prawn, Macrobrachium rosenbergii was used as test animal and the experiment was lost for 112 days. Significantly, higher gain in weight of prawn was recorded in the treatment with 6.07% sardine oil incorporation followed by higher specific growth rate, food conversion rate and protein efficiency ratio. The 8.07% sardine oil diet produced the highest RNA: DNA ratio in the prawn muscle. Digestive enzyme analyses in the digestive tract and mid-gut gland showed the greatest activity in prawns fed the 8.07% diet.

Keywords: digestive enzyme, fish diet, Macrobrachium rosenbergii, sardine oil

Procedia PDF Downloads 320

Authors: Paula K. Novelli, Margarida M. Barros, Luciana F. Flueri

Abstract:

Utilization of agricultural by-products, wheat ban and soybean bran, as substrate for solid state fermentation (SSF) was studied, aiming the achievement of different enzymes from Aspergillus sp. with distinct biological characteristics and its application and improvement on animal nutrition. Aspergillus niger and Aspergillus oryzea were studied as they showed very high yield of phytase and protease production, respectively. Phytase activity was measure using p-nitrophenilphosphate as substrate and a standard curve of p-nitrophenol, as the enzymatic activity unit was the quantity of enzyme necessary to release one μmol of p-nitrophenol. Protease activity was measure using azocasein as substrate. Activity for phytase and protease substantially increased when the different biochemical characteristics were considered in the study. Optimum pH and stability of the phytase produced by A. niger with wheat bran as substrate was between 4.0 - 5.0 and optimum temperature of activity was 37oC. Phytase fermented in soybean bran showed constant values at all pHs studied, for optimal and stability, but low production. Phytase with both substrates showed stable activity for temperatures higher than 80oC. Protease from A. niger showed very distinct behavior of optimum pH, acid for wheat bran and basic for soybean bran, respectively and optimal values of temperature and stability at 50oC. Phytase produced by A. oryzae in wheat bran had optimum pH and temperature of 9 and 37oC, respectively, but it was very unstable. On the other hand, proteases were stable at high temperatures, all pH’s studied and showed very high yield when fermented in wheat bran, however when it was fermented in soybean bran the production was very low. Subsequently the upscale production of phytase from A. niger and proteases from A. oryzae were applied as an enzyme additive in fish fed for digestibility studies. Phytases and proteases were produced with stable enzyme activity of 7,000 U.g-1 and 2,500 U.g-1, respectively. When those enzymes were applied in a plant protein based fish diet for digestibility studies, they increased protein, mineral, energy and lipids availability, showing that these new enzymes can improve animal production and performance. In conclusion, the substrate, as well as, the microorganism species can affect the biochemical character of the enzyme produced. Moreover, the production of these enzymes by SSF can be up to 90% cheaper than commercial ones produced with the same fungi species but submerged fermentation. Add to that these cheap enzymes can be easily applied as animal diet additives to improve production and performance.

Keywords: agricultural by-products, animal nutrition, enzymes production, solid state fermentation

Procedia PDF Downloads 321

Authors: Yomna Ibrahim El-Gazzar, Hussien Ibrahim El-Subbagh, Hanan Hanaa Georgey, Ghada S. Hassan Hassan

Abstract:

Dihydrofolate reductase (DHFR) is an enzyme that has pivotal importance in biochemistry and medicinal chemistry. It catalyzes the reduction of dihydrofolate to tetrahydrofolate and intimately couples with thymidylate synthase. Thymidylate synthase is a crucial enzyme that catalyzes the reductive methylation of (dUMP) to (dTMP) utilizing N5, N10-methylenetetrahydrofolate as a cofactor. A new series of 2-substituted thio-quinazolin-4-one analogs was designed that possessed electron withdrawing or donating functional groups (Cl or OCH3) at position 6- or 7-, 4-methoxyphenyl function at position 3-.The thiol function is used to connect to either 1,2,4-triazole, or 1,3,4-thiadiazole via a methylene bridge. Most of the functional groups designed to be accommodated on the quinazoline ring such as thioether, alkyl to increase lipid solubility of polar compounds, a character very much needed in the nonclassical DHFR inhibitors. The target compounds were verified with spectral data and elemental analysis. DHFR inhibitions, as well as antitumor activity, were applied on three cell lines (MCF-7, CACO-2, HEPG-2).

Keywords: nonclassical antifolates, DHFR Inhibitors, antitumor activity, quinazoline ring

Procedia PDF Downloads 385

Authors: Archana. S, Usha Rani. G, Anand Balakrishnan, Sanjana.R, Solomon F, Vijayalakshmi. J

Abstract:

Background: There is evidence that male factors contribute to the infertility of up to 50% of couples, who are evaluated and treated for infertility using advanced assisted reproductive technologies. Genetic abnormalities, including sperm chromosome aneuploidy as well as structural aberrations, are one of the major causes of male infertility. Recent advances in technology expedite the evaluation of sperm aneuploidy. The purpose of the study was to de-termine the prevalence of sperm aneuploidy in infertile males and the degree of association between DNA fragmentation and sperm aneuploidy. Methods: In this study, 75 infertile men were included, and they were divided into four abnormal groups (Oligospermia, Terato-spermia, Asthenospermia and Oligoasthenoteratospermia (OAT)). Men with children who were normozoospermia served as the control group. The Fluorescence in situ hybridization (FISH) method was used to test for sperm aneuploidy, and the Sperm Chromatin Dispersion Assay (SCDA) was used to measure the fragmentation of sperm DNA. Spearman's correla-tion coefficient was used to evaluate the relationship between sperm aneuploidy and sperm DNA fragmentation along with age. P < 0.05 was regarded as significant. Results: 75 partic-ipants' ages varied from 28 to 48 years old (35.5±5.1). The percentage of spermatozoa bear-ing X and Y was determined to be statistically significant (p-value < 0.05) and was found to be 48.92% and 51.18% of CEP X X 1 – nucish (CEP XX 1) [100] and CEP Y X 1 – nucish (CEP Y X 1) [100]. When compared to the rate of DNA fragmentation, it was discovered that infertile males had a greater frequency of sperm aneuploidy. Asthenospermia and OAT groups in sex chromosomal aneuploidy were significantly correlated (p<0.05). Conclusion: Sperm FISH and SCDA assay results showed increased sperm aneuploidy frequency, and DNA fragmentation index in infertile men compared with fertile men. There is a significant relationship observed between sperm aneuploidy and DNA fragmentation in OAT patients. When evaluating male variables and idiopathic infertility, the sperm FISH screening method can be used as a valuable diagnostic tool.

Keywords: ale infertility, dfi (dna fragmentation assay) (scd-sperm chromatin dispersion).art (artificial reproductive technology), trisomy, aneuploidy, fish (fluorescence in-situ hybridization), oat (oligoasthoteratospermia)

Procedia PDF Downloads 47

Authors: Tooba N. Shamsi, Romana Perveen, Sadaf Fatima

Abstract:

Protease Inhibitors (PIs) are widespread in nature, produced by animals, plants and microorganisms. They play vital role in various biological activities by keeping a check on activity of proteases. Present study aims to investigate antioxidant and anti-inflammatory properties of PPI from Cajanus cajan (CCTI) and Phaseolus limensis (LBTI). PPI was purified from C. cajan (PUSA-992) by ammonium sulfate precipitation followed by ion exchange chromatography. The anti-oxidant activity was analyzed by two most common radical scavenging assays of FRAP (ferric reducing antioxidant power) and DPPH (1,1- diphenyl-2-picrylhydrazyl). Also, in-vitro anti-inflammatory activity was evaluated using albumin denaturation assay and membrane stabilization assay at different concentrations. Ascorbic acid and aspirin were used as a standards for antioxidant and anti-inflammatory assays respectively. The PPIs were also checked for antimicrobial activity against a number of bacterial strains. The CCTI and LBTI showed DPPH radical scavenging activity in a concentration–dependent manner with IC50 values 544 µg/ml and 506 µg/ml respectively comparative to ascorbic acid which was 258 µg/ml. Following FRAP assay, it was evaluated that LBTI had 87.5% and CCTI showed 84.4% antioxidant activity, taking value of standard ascorbic acid to be 100%. The PPIs also showed in-vitro anti‐inflammatory activity by inhibiting the heat induced albumin denaturation with IC50 values of 686 µg/ml and 615 µg/ml for CCTI and LBTI respectively compared to the standard (aspirin) which was 70.8 µg/ml. Red blood cells membrane stabilization with IC50 values of 641 µg/ml and 587 µg/ml for CCTI and LBTI respectively against aspirin which showed IC50 value of 70.4 µg/ml. PPIs showed antibacterial activity against 7 known strains while there was apparently no action against fungi.

Keywords: Cajanus cajan, Phaseolus limensis, Lima beans, protein protease inhibitor, antioxidant, anti-inflammatory, antimicrobial activity

Procedia PDF Downloads 288

Authors: Mahsa Taghavi

Abstract:

In the treatment of breast cancer, tamoxifen is a regularly prescribed medication. The effect of tamoxifen on breast cancer patients' EMT pathways was studied. In this study to see if it had any effect on the cancer cells' resistance to tamoxifen and to look for specific miRNAs associated with EMT. In this work, we used continuous and integrated bioinformatics analysis to choose the optimal GEO datasets. Once we had sorted the gene expression profile, we looked at the mechanism of signaling, the ontology of genes, and the protein interaction of each gene. In the end, we used the GEPIA database to confirm the candidate genes. after that, I investigated critical miRNAs related to candidate genes. There were two gene expression profiles that were categorized into two distinct groups. Using the expression profile of genes that were lowered in the EMT pathway, the first group was examined. The second group represented the polar opposite of the first. A total of 253 genes from the first group and 302 genes from the second group were found to be common. Several genes in the first category were linked to cell death, focal adhesion, and cellular aging. Two genes in the second group were linked to cell death, focal adhesion, and cellular aging. distinct cell cycle stages were observed. Finally, proteins such as MYLK, SOCS3, and STAT5B from the first group and BIRC5, PLK1, and RAPGAP1 from the second group were selected as potential candidates linked to tamoxifen's influence on the EMT pathway. hsa-miR-1204 and hsa-miR-639 have a very close relationship with the candidates genes according to the node degrees and betweenness index. With this, the action of tamoxifen on the EMT pathway was better understood. It's important to learn more about how tamoxifen's target genes and proteins work so that we can better understand the drug.

Keywords: tamoxifen, breast cancer, bioinformatics analysis, EMT, miRNAs

Procedia PDF Downloads 126

Authors: Mohammad Shamim Hasan Mandal, Phu Minh, Do Tan Khang, Phung Thi Tuyen, Tran Dang Xuan

Abstract:

Rice (Oryza sativa L.) is one of the major crops of Vietnam which has more than thousands of varieties. Many of the local varieties have greater potentiality but they are in danger of extinct. Rice plant contains many secondary metabolites that are allelopathic to other plants. Seven rice varieties were cultivated in the field condition at Hiroshima University, Japan; stems and leaves from each variety were collected later, they were extracted with methanol, hexane, ethyl acetate, butanol, and water. Total phenolic content and total flavonoid contents were high in ethyl acetate extracts. DPPH antioxidant assay results showed that the ethyl acetate extracts had the higher IC50 value. Therefore, the ethyl acetate extracts were selected for laboratory experimentation through petri dish assay. Results showed that the two-local variety Re nuoc and Nan chon completely inhibited the germination of radish seedlings. Further laboratory bioassay and field experimentation will be conducted to validate the laboratory bioassay findings.

Keywords: allelopathy, bioassay, Oryza sativa, Raphanus sativus

Procedia PDF Downloads 355

Authors: Simona Moravcova, Jiri Novotny, Zdenka Bendova

Abstract:

The pineal gland of the vertebrate brain is a circumventricular organ which serves as a major neuroendocrine gland with the primary function of rhythmic secretion of neurohormone melatonin under the control of the hypothalamic suprachiasmatic nucleus (SCN). Soon after the onset of the darkness, the activity of the key rate-limiting enzyme for melatonin synthesis, arylalkylamine N-acetyltransferase (AANAT), raises due to the increased release of norepinephrine from sympathetic neurons terminating on the parenchymal cells where it binds to β-adrenergic receptors. Melatonin codes the length of the night, and it is well recognized for its anti-inflammatory effects. However, to our knowledge, less is known about the effect of the immune system on the melatonin biosynthesis and the precise role of the STAT3 in the signaling pathway leading to the expression of AANAT. Lipopolysaccharide (LPS) is the essential component in the outer surface membrane of gram-negative bacteria and acts as a strong stimulator of natural and innate immunity. STAT3 acts as an important factor in immune response. Here we investigated the effect of LPS on the components of the melatonergic synthetic pathway in the pineal gland. The experiments were performed both in vivo and in vitro. The changes in AANAT activity were determined by radioenzymatic assay. PCR analyses were carried out to detect aa-nat, icer, spi-3 and stat3 gene expression. From our results, it is apparent that the high basal level of phosphorylated forms of STAT3 can be elevated after systemic as well as in vitro administration of LPS. Our experiments have shown that LPS reduces melatonin synthesis, nevertheless, the activity of AANAT was increased. Moreover, the basal level of phosphorylated STAT3 counteracts β-adrenergic receptor-mediated aa-nat gene expression and sustains its own and spi-3 gene expression. In conclusion, LPS can affect immunomodulators such as melatonin in the pineal gland.

Keywords: AANAT, lipopolysaccharide, pineal gland, rat, STAT3

Procedia PDF Downloads 166

Authors: Noura El-Ahmady El-Naggar

Abstract:

Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

Procedia PDF Downloads 357

Authors: Jacky Bhagat, Baban S Ingole

Abstract:

In this study, in vivo experiments were carried out to investigate the effects of a toxic polycyclic aromatic hydrocarbon (PAH), benzo(k)fluoranthene (B[k]F), on marine gastropod, Morula granulata collected from Goa, west coast of India. Snails were exposed to different concentrations of B(k)F (1, 10, 25 and 50 µg/L) for 96 h. The genotoxic effects were evaluated by measuring DNA strand breaks using alkaline comet assay and oxidative stress were measured with the help of battery of biomarkers such as superoxide dismutase (SOD) catalase (CAT), glutathione-s-transferase (GST), and lipid peroxidation (LPO). Concentration-dependent increase in percentage tail DNA (TDNA) was observed in snails exposed to B(k)F. Exposure concentrations above 1 µg/L of B(k)F, showed significant increase in SOD activity and LPO value in snails. After 96 h, SOD activity were found to be doubled for 50 µg/L of B(k)F with reference to control. Significant increase in CAT and GST activity was observed at all exposure conditions at the end of the exposure time. Our study showed that B(k)F induces oxidative stress in snails which further lead to genotoxic damage.

Keywords: benzo(k)fluoranthene, comet assay, gastropod, oxidative stress

Procedia PDF Downloads 335

Authors: Jose Gonzalez, Fai Cheng, Ivy Fan

Abstract:

Shipping has a mature and ever expanding worldwide market. The future of the marine industry itself is not only irrevocably linked with the global economic, social, and political landscape; it is also subject to the technological developments in different fields. Some of them may have never been linked to the marine industry before. Companies in the marine sector are getting more dependent on technologies to achieve competitive advantage in an increasing open market. Technologies can be fused across different business functions and geopolitical influences. A successful marine business should be prepared to embrace such potential changes that lie ahead. The present paper intends to articulate long-term marine technology strategies from an industrial perspective. Methodology and current development are introduced. The paper will also provide insight into future technological trends demand for major commercial ship types. It may also assist different stakeholders in tailoring their long-term strategies to achieve a Sea Change and to uncap opportunity.

Keywords: commercial sector, marine, trends, technology

Procedia PDF Downloads 401