Search results for: plasma protein electrophoresis
1663 Inhibition of Glutamate Carboxypeptidase Activity Protects Retinal Ganglionic Cell Death Induced by Ischemia-Reperfusion by Reducing the Astroglial Activation in Rat
Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo
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Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell
Procedia PDF Downloads 3401662 Determination of the Element Contents in Turkish Coffee and Effect of Sugar Addition
Authors: M. M. Fercan, A. S. Kipcak, O. Dere Ozdemir, M. B. Piskin, E. Moroydor Derun
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Coffee is a widely consumed beverage with many components such as caffeine, flavonoids, phenolic compounds, and minerals. Coffee consumption continues to increase due to its physiological effects, its pleasant taste, and aroma. Robusta and Arabica are two basic types of coffee beans. The coffee bean used for Turkish coffee is Arabica. There are many elements in the structure of coffee and have various effect on human health such as Sodium (Na), Boron (B), Magnesium (Mg) and Iron (Fe). In this study, the amounts of Mg, Na, Fe, and B contents in Turkish coffee are determined and effect of sugar addition is investigated for conscious consumption. The analysis of the contents of coffees was determined by using inductively coupled plasma optical emission spectrometry (ICP-OES). From the results of the experiments the Mg, Na, Fe and B contents of Turkish coffee after sugar addition were found as 19.83, 1.04, 0.02, 0.21 ppm, while without using sugar these concentrations were found 21.46, 0.81, 0.008 and 0.16 ppm. In addition, element contents were calculated for 1, 3 and 5 cups of coffee in order to investigate the health effects.Keywords: health effect, ICP-OES, sugar, Turkish coffee
Procedia PDF Downloads 4981661 Home Made Rice Beer Waste (Choak): A Low Cost Feed for Sustainable Poultry Production
Authors: Vinay Singh, Chandra Deo, Asit Chakrabarti, Lopamudra Sahoo, Mahak Singh, Rakesh Kumar, Dinesh Kumar, H. Bharati, Biswajit Das, V. K. Mishra
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The most widely used feed resources in poultry feed, like maize and soybean, are expensive as well as in short supply. Hence, there is a need to utilize non-conventional feed ingredients to cut down feed costs. As an alternative, brewery by-products like brewers’ dried grains are potential non-conventional feed resources. North-East India is inhabited by many tribes, and most of these tribes prepare their indigenous local brew, mostly using rice grains as the primary substrate. Choak, a homemade rice beer waste, is an excellent and cheap source of protein and other nutrients. Fresh homemade rice beer waste (rice brewer’s grain) was collected locally. The proximate analysis indicated 28.53% crude protein, 92.76% dry matter, 5.02% ether extract, 7.83% crude fibre, 2.85% total ash, 0.67% acid insoluble ash, 0.91% calcium, and 0.55% total phosphorus. A feeding trial with 5 treatments (incorporating rice beer waste at the inclusion levels of 0,10,20,30 & 40% by replacing maize and soybean from basal diet) was conducted with 25 laying hens per treatment for 16 weeks under completely randomized design in order to study the production performance, blood-biochemical parameters, immunity, egg quality and cost economics of laying hens. The results showed substantial variations (P<0.01) in egg production, egg mass, FCR per dozen eggs, FCR per kg egg mass, and net FCR. However, there was not a substantial difference in either body weight or feed intake or in egg weight. Total serum cholesterol reduced significantly (P<0.01) at 40% inclusion of rice beer waste. Additionally, the egg haugh unit grew considerably (P<0.01) when the graded levels of rice beer waste increased. The inclusion of 20% rice brewers dried grain reduced feed cost per kg egg mass and per dozen egg production by Rs. 15.97 and 9.99, respectively. Choak (homemade rice beer waste) can thus be safely incorporated into the diet of laying hens at a 20% inclusion level for better production performance and cost-effectiveness.Keywords: choak, rice beer waste, laying hen, production performance, cost economics
Procedia PDF Downloads 591660 Antioxidant Capacity of Maize Corn under Drought Stress from the Different Zones of Growing
Authors: Astghik R. Sukiasyan
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The semidental sweet maize of Armenian population under drought stress and pollution by some heavy metals (HMs) in sites along the river Debet was studied. Accordingly, the objective of this work was to investigate the antioxidant status of maize plant in order to identify simple and reliable criteria for assessing the degree of adaptation of plants to abiotic stress of drought and HMs. It was found that in the case of removal from the mainstream of the river, the antioxidant status of the plant varies. As parameters, the antioxidant status of the plant has been determined by the activity of malondialdehyde (MDA) and Ferric Reducing Ability of Plasma (FRAP), taking into account the characteristics of natural drought of this region. The possibility of using some indicators which characterized the antioxidant status of the plant was concluded. The criteria for assessing the extent of environmental pollution could be HMs. This fact can be used for the early diagnosis of diseases in the population who lives in these areas and uses corn as the main food.Keywords: antioxidant status, maize corn, drought stress, heavy metal
Procedia PDF Downloads 2701659 Dietary Diversification and Nutritional Education: A Strategy to Improve Child Food Security Status in the Rural Mozambique
Authors: Rodriguez Diego, Del Valle Martin, Hargreaves Matias, Riveros Jose Luis
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Nutrient deficiencies due to a diet low in quantitative and qualitative terms, are prevalent throughout the developing world, especially in sub-Saharan Africa. Children and women of childbearing age are especially vulnerable. Limited availability, access and intake of animal foods at home and lack of knowledge about their value in the diet and the role they play in health, contribute to poor diet quality. Poor bioavailability of micronutrients in diets based on foods high in fiber and phytates, the low content of some micronutrients in these foods are further factors to consider. Goats are deeply embedded in almost every Sub-Saharan African rural culture, generally kept for their milk, meat, hair or leather. Goats have played an important role in African social life, especially in food security. Goat meat has good properties for human wellbeing, with a special role in lower income households. It has a high-quality protein (20 protein g/100 meat g) including all essential amino acids, good unsaturated/satured fatty acids relationship, and it is an important B-vitamin source with high micronutrients bioavailability. Mozambique has major food security problems, with poor food access and utilization, undiversified diets, chronic poverty and child malnutrition. Our objective was to design a nutritional intervention based on a dietary diversification, nutritional education, cultural beliefs and local resources, aimed to strengthen food security of children at Barrio Broma village (15°43'58.78"S; 32°46'7.27"E) in Chitima, Mozambique. Two surveys were conducted first of socio-productive local databases and then to 100 rural households about livelihoods, food diversity and anthropometric measurements in children under 5 years. Our results indicate that the main economic activity is goat production, based on a native breed with two deliveries per year in the absence of any management. Adult goats weighted 27.2±10.5 kg and raised a height of 63.5±3.8 cm. Data showed high levels of poverty, with a food diversity score of 2.3 (0-12 points), where only 30% of households consume protein and 13% iron, zinc, and B12 vitamin. The main constraints to food security were poor access to water and low income to buy food. Our dietary intervention was based on improving diet quality by increasing the access to dried goat meat, fresh vegetables, and legumes, and its utilization by a nutritional education program. This proposal was based on local culture and living conditions characterized by the absence of electricity power and drinkable water. The drying process proposed would secure the food maintenance under local conditions guaranteeing food safety for a longer period. Additionally, an ancient local drying technique was rescued and used. Moreover, this kind of dietary intervention would be the most efficient way to improve the infant nutrition by delivering macro and micronutrients on time to these vulnerable populations.Keywords: child malnutrition, dietary diversification, food security, goat meat
Procedia PDF Downloads 3021658 Square Wave Anodic Stripping Voltammetry of Copper (II) at the Tetracarbonylmolybdenum(0) MWCNT Paste Electrode
Authors: Illyas Isa, Mohamad Idris Saidin, Mustaffa Ahmad, Norhayati Hashim
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A highly selective and sensitive electrode for determination of trace amounts of Cu (II) using square wave anodic stripping voltammetry (SWASV) was proposed. The electrode was made of the paste of multiwall carbon nanotubes (MWCNT) and 2,6–diacetylpyridine-di-(1R)–(-)–fenchone diazine tetracarbonylmolybdenum(0) at 100:5 (w/w). Under optimal conditions the electrode showed a linear relationship with concentration in the range of 1.0 × 10–10 to 1.0 × 10– 6 M Cu (II) and limit of detection 8.0 × 10–11 M Cu (II). The relative standard deviation (n = 5) of response to 1.0 × 10–6 M Cu(II) was 0.036. The interferences of cations such as Ni(II), Mg(II), Cd(II), Co(II), Hg(II), and Zn(II) (in 10 and 100-folds concentration) are negligible except from Pb (II). Electrochemical impedance spectroscopy (EIS) showed that the charge transfer at the electrode-solution interface was favorable. Result of analysis of Cu(II) in several water samples agreed well with those obtained by inductively coupled plasma-optical emission spectrometry (ICP-OES). The proposed electrode was then recommended as an alternative to spectroscopic technique in analyzing Cu (II).Keywords: chemically modified electrode, Cu(II), Square wave anodic stripping voltammetry, tetracarbonylmolybdenum(0)
Procedia PDF Downloads 2621657 LncRNA-miRNA-mRNA Networks Associated with BCR-ABL T315I Mutation in Chronic Myeloid Leukemia
Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang
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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation
Procedia PDF Downloads 1591656 Inflammatory Changes Caused by Lipopolysaccharide in Odontoblasts
Authors: Virve Pääkkönen, Heidi M. Cuffaro, Leo Tjäderhane
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Objectives: Odontoblasts are the outermost cell layer of dental pulp and form the dentin. Importance of bacterial products, e.g. lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria and lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, have been indicated in the pathogenesis of pulpitis. Gram-positive bacteria are more prevalent in superficial carious lesion while the amount gram-negative is higher in the deep lesions. Objective of this study was to investigate the effect of these bacterial products on inflammatory response of pulp tissue. Interleukins (IL) were of special interest. Various ILs have been observed in the dentin-pulp complex of carious tooth in vivo. Methods: Tissue culture method was used for testing the effect of LTA and LPS on human odontoblasts. Enzymatic isolation technique was used to extract living odontoblasts for cell cultures. DNA microarray and quantitative PCR (qPCR) were used to characterize the changes in the expression profile of the tissue cultured odontoblasts. Laser microdissection was used to cut healthy and affected dentin and odontoblast layer directly under carious lesion for experiments. Cytokine array detecting 80 inflammatory cytokines was used to analyze the protein content of conditioned culture media as well as dentin and odontoblasts from the carious teeth. Results: LPS caused increased gene expression IL-1α, and -8 and decrease of IL-1β, 12 , -15 and -16 after 1h treatment, while after 24h treatment decrease of IL-8, -11 and 23 mRNAs was observed. LTA treatment caused cell death in the tissue cultured odontoblasts but in in the cell culture but not in cell culture. Cytokine array revealed at least 2-fold down-regulation of IL-1β, -10 and -12 in response to LPS treatment. Cytokine array of odontoblasts of carious teeth, as well as LPS-treated tissue-cultured odontoblasts, revealed increased protein amounts of IL-16, epidermal growth factor (EGF), angiogenin and IGFBP-1 as well as decreased amount of fractalkine. In carious dentin, increased amount of IL-1β, EGF and fractalkine was observed, as well as decreased level of GRO-1 and HGF. Conclusion: LPS caused marked changes in the expression of inflammatory cytokines in odontoblasts. Similar changes were observed in the odontoblasts cut directly under the carious lesion. These results help to shed light on the inflammatory processes happening during caries.Keywords: inflammation, interleukin, lipoteichoic acid, odontoblasts
Procedia PDF Downloads 2111655 Development of High Temperature Eutectic Oxide Ceramic Matrix Composites
Authors: Yağmur Can Gündoğan, Kübra Gürcan Bayrak, Ece Özerdem, Buse Katipoğlu, Erhan Ayas, Rifat Yılmaz
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Eutectic oxide based ceramic matrix composites have a unique microstructure that does not include grain boundary in the form of a continuous network. Because of this, these materials have the properties of perfect high-temperature strength, creep strength, and high oxidation strength. Mechanical properties of them are much related to occurring solidification structures during eutectic reactions. One of the most important production methods of this kind of material is the process of vacuum arc melting. Within scope of this studying, it is aimed to investigate the production of Al₂O₃-YAG-based eutectic ceramics by Arc melting and Spark Plasma Sintering methods for use in aerospace and defense industries where high-temperature environments play an important role and to examine the effects of ZrO₂ and LiF additions on microstructure development and mechanical properties.Keywords: alumina, composites, eutectic, YAG
Procedia PDF Downloads 1181654 Effect of Environmental Stress Factors on the Degradation of Display Glass
Authors: Jinyoung Choi, Hyun-A Kim, Sunmook Lee
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The effects of environmental stress factors such as storage conditions on the deterioration phenomenon and the characteristic of the display glass were studied. In order to investigate the effect of chemical stress on the glass during the period of storage, the respective components of commercial glass were first identified by XRF (X-ray fluorescence). The glass was exposed in the acid, alkali, neutral environment for about one month. Thin film formed on the glass surface was analyzed by XRD (X-ray diffraction) and FT-IR (Fourier transform infrared). The degree of corrosion and the rate of deterioration of each sample were confirmed by measuring the concentrations of silicon, calcium and chromium with ICP-OES (Inductively coupled plasma-optical emission spectrometry). The optical properties of the glass surface were confirmed by SEM (Scanning electron microscope) before and after the treatment. Acknowledgement—The authors gratefully acknowledge the financial support from the Ministry of Trade, Industry and Energy (Grant Number: 10076817)Keywords: corrosion, degradation test, display glass, environmental stress factor
Procedia PDF Downloads 4591653 Polyclonal IgG glycosylation in Patients with Pediatric Appendicitis
Authors: Dalma Dojcsák, Csaba Váradi, Flóra Farkas, Tamás Farkas, János Papp, Béla Viskolcz
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Background: Appendicitis is a common acute inflammatory condition in both children and adults, but current laboratory markers such as C-reactive protein (CRP), white blood cell count (WBC), absolute neutrophil count (ANC), and red blood cell count (RNC) lack specificity in detecting appendicitis-related inflammation. N-glycosylation, an asparagine-linked glycosylation process, plays a vital role in cellular interactions, angiogenesis, immune response, and effector functions. Altered N-glycosylation impacts tumor growth and both acute and chronic inflammatory processes. IgG, the second most abundant glycoprotein in serum, shows altered glycosylation patterns during inflammation, suggesting that IgG glycan modifications may serve as potential biomarkers for appendicitis. Specifically, increased levels of agalactosylated IgG glycans are a known feature of various inflammatory conditions, potentially including appendicitis. Identifying pediatric appendicitis remains challenging due to the absence of specific biomarkers, which makes diagnosis reliant on clinical symptoms, imaging such as ultrasound, and nonspecific lab indicators (e.g., CRP, WBC, ANC). In this study, we analyzed the IgG derived N-glycome in pediatric patients with appendicitis compared with healthy controls. Methodology: The N-glycome was analyzed by high-performance liquid-chromatography combined with mass spectrometry. IgG was isolated from serum samples by Protein G column. The IgG derived glycans were released by enzymatic deglycosylation and fluorescent tags were attached to each glycan moiety, which made necessitates the sample clean-up for further reliable quantitation. Overall, 38 controls and 40 serum samples diagnosed with pediatric appendicitis were analyzed by HILIC-MS methods. Multivariate statistical tests were performed with area percentage under the peak data derived from the integrated peaks, which were obtained from the chromatograms. Conclusions: Our results represented the altered N-glycome of IgG in pediatric appendicitis is similar with other observations. The glycosylation pattern reported so far for IgG is characterized by decreased galactosylation and sialylation, and an increase in fucosylation.Keywords: N-glycosylation, liquid chromatography, mass spectrometry, inflammation, appendicitis, immunoglobulin G
Procedia PDF Downloads 111652 Quantitative Elemental Analysis of Cyperus rotundus Medicinal Plant by Particle Induced X-Ray Emission and ICP-MS Techniques
Authors: J. Chandrasekhar Rao, B. G. Naidu, G. J. Naga Raju, P. Sarita
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Particle Induced X-ray Emission (PIXE) and Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) techniques have been employed in this work to determine the elements present in the root of Cyperus rotundus medicinal plant used in the treatment of rheumatoid arthritis. The elements V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Rb, and Sr were commonly identified and quantified by both PIXE and ICP-MS whereas the elements Li, Be, Al, As, Se, Ag, Cd, Ba, Tl, Pb and U were determined by ICP-MS and Cl, K, Ca, Ti and Br were determined by PIXE. The regional variation of elemental content has also been studied by analyzing the same plant collected from different geographical locations. Information on the elemental content of the medicinal plant would be helpful in correlating its ability in the treatment of rheumatoid arthritis and also in deciding the dosage of this herbal medicine from the metal toxicity point of view. Principal component analysis and cluster analysis were also applied to the data matrix to understand the correlation among the elements.Keywords: PIXE, CP-MS, elements, Cyperus rotundus, rheumatoid arthritis
Procedia PDF Downloads 3331651 Unlocking the Health Benefits of Goat Meat
Authors: K. Makangali, G. Tokysheva, A. Shoman
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Goat meat and goat meat products have garnered increasing attention within the realm of nutrition and health due to their potential to provide a myriad of benefits. This scientific article presents a comprehensive review of the health advantages associated with goat meat consumption and the products derived from it. The paper explores the nutritional content of goat meat, highlighting its favorable composition in terms of protein, essential minerals, and amino acids. It delves into the intricate balance of macronutrients, with lower fat and cholesterol levels compared to other meats, making goat meat a desirable choice for individuals seeking healthier dietary options.Keywords: goat meat, amino acid, nutrition, meat products, meat
Procedia PDF Downloads 791650 A Turn-on Fluorescent Sensor for Pb(II)
Authors: Ece Kök Yetimoğlu, Soner Çubuk, Neşe Taşci, M. Vezir Kahraman
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Lead(II) is one of the most toxic environmental pollutants in the world, due to its high toxicity and non-biodegradability. Lead exposure causes severe risks to human health such as central brain damages, convulsions, kidney damages, and even death. To determine lead(II) in environmental or biological samples, scientists use atomic absorption spectrometry (AAS), inductively coupled plasma mass spectrometry (ICPMS), fluorescence spectrometry and electrochemical techniques. Among these systems the fluorescence spectrometry and fluorescent chemical sensors have attracted considerable attention because of their good selectivity and high sensitivity. The fluorescent polymers usually contain covalently bonded fluorophores. In this study imidazole based UV cured polymeric film was prepared and designed to act as a fluorescence chemo sensor for lead (II) analysis. The optimum conditions such as influence of pH value and time on the fluorescence intensity of the sensor have also been investigated. The sensor was highly sensitive with a detection limit as low as 1.87 × 10−8 mol L-1 and it was successful in the determination of Pb(II) in water samples.Keywords: fluorescence, lead(II), photopolymerization, polymeric sensor
Procedia PDF Downloads 6711649 Genome Analyses of Pseudomonas Fluorescens b29b from Coastal Kerala
Authors: Wael Ali Mohammed Hadi
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Pseudomonas fluorescens B29B, which has asparaginase enzymatic activity, was isolated from the surface coastal seawater of Trivandrum, India. We report the complete Pseudomonas fluorescens B29B genome sequenced, identified, and annotated from a marine source. We find the genome at most minuscule a 7,331,508 bp single circular chromosome with a GC content of 62.19% and 6883 protein-coding genes. Three hundred forty subsystems were identified, including two predicted asparaginases from the genome analysis of P. fluorescens B29B for further investigation. This genome data will help further industrial biotechnology applications of proteins in general and asparaginase as a target.Keywords: pseudomonas, marine, asparaginases, Kerala, whole-genome
Procedia PDF Downloads 2151648 Determination of Some Biochemical Values for the Liza klunzingeri in Coastal Water of Persian Gulf
Authors: Majid Afkhami, Maryam Ehsanpour
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Serum biochemical can be used for monitoring any changes in the physiological condition of fish and quality of waters. The aim of this paper was to determine of plasma sugar, triglycerides, cholesterol, iron, ALP (alkaline phosphatase) and LDH (lactate dehydrogenase) levels of Liza klunzingeri in Persian Gulf. Blood sample was collected from the caudal vessel with syringes coated with sodium heparin. Biochemical values were: sugar 110.37±28.46 mg/di, triglycerides 96.82±23.40 mg/di, cholesterol 177.28 ±40.75 mg/di, iron 104.74± 19.08 mg/di, ALP 117.62±34.49 u/l, LDH 1613.00±345.34 u/l. A significant positive correlation (P<0.01) was found between triglycerides and sugar. Triglycerides had a significant and positive relationship with cholesterol (P<0.01). ALP also had a significant and positive relationship with sugar (P<0.01) and triglycerides (P<0.05). LDH correlated positively with sugar, cholesterol, triglycerides (P<0.01) and ALP (P<0.05). The results revealed reverse correlation between iron with cholesterol, sugar, triglycerides, ALP, and LDH (P<0.01). This study represents a contribution to the referential biochemical values of the L. klunzingeri. In further studies, the established reference ranges might be useful for the health assessment of this species.Keywords: Liza klunzingeri, blood, ALP, LDH
Procedia PDF Downloads 6251647 Targeting Matrix Metalloprotease-9 to Reduce Coronary Artery Manifestations of Kawasaki’s Disease
Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian
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Kawasaki disease (KD) is the primary cause of acquired pediatric heart disease as an acute vasculitis. In children with prolonged fever, rash, and inflammation of the mucosa KD must be considered as a clinical diagnosis. There is a persuasive suggestion of immune-mediated damage as the pathophysiologic cascade of KD. For example, the invasion of cytotoxic T-cells supports a viral etiology and the inflammasome of the innate immune system is a critical component in the vasculitis formation in KD. Animal models of KD propose the cytokine profiles, such as increased IL-1 and GM-CSF, which cause vascular damage. CRP and IFN-γ elevated expression and the upregulation of IL-6, and IL-10 production are also described in previous studies. Untreated KD is a critical risk factor for coronary artery diseases and myocardial infarction. Vascular damage may encompass amplified T-cell activity. SMAD3 is an essential molecule in down-regulating T-cells and increasing expression of FoxP3. It has a critical effect in the differentiation of regulatory T-cells. The discrepancy of regulatory T-cells and pro-inflammatory Th17 has been studied in acute coronary syndrome during KD. However in the coronary artery damaged lymphocytes and IgA plasma cells are seen at the lesion locations, the major immune cells in the coronary lesions are monocytes/macrophages and neutrophils. These cells secrete TNF-α, and activates matrix metalloprotease (MMP)-9, reducing the integrity of vessels and prompting patients to arise aneurysm. MMPs can break down the components of the extracellular matrix and assist immune cell movement. IVIG as an effective form of treatment clarified the role of the immune system, which may target pathogenic antigens and regulate cytokine production. Several reports have revealed that in the coronary arteries, high expression of MMP-9 in monocyte/macrophage results in pathologic cascades. Curcumin is a potent antioxidant and anti-inflammatory molecule. Curcumin decreases the production of reactive oxygen and nitrogen species and inhibits transcription factors like AP-1 and NF-κB. Curcumin also contains the characteristics of inhibitory effects on MMPs, especially MMP-9. The upregulation of MMP-9 is an important cellular response. Curcumin treatment caused a reverse effect and down-regulates MMP-9 gene expression which may fund the anti-inflammatory effect. Curcumin inhibits MMP-9 expression via PKC and AMPK-dependent pathways in Human monocytes cells. Elevated expression and activity of MMP-9 are correlated with advanced vascular lesions. AMPK controls lipid metabolism and oxidation, and protein synthesis. AMPK is also necessary for the MMP-9 activity and THP-1 cell adhesion to endothelial cells. Curcumin was shown to inhibit the activation of AMPKα. Compound C (AMPK inhibitor) inhibits MMP-9 expression level. Therefore, through inactivating AMPKs and PKC, curcumin decreases the MMP-9 level, which results in inhibiting monocyte/macrophage differentiation. Compound C also suppress the phosphorylation of three major classes of MAP kinase signaling, suggesting that curcumin may suppress MMP-9 level by inactivation of MAPK pathways. MAPK cascades are activated to induce the expression of MMP-9. Curcumin inhibits MAPKs phosphorylation, which contributes to the down-regulation of MMP-9. This study demonstrated that the potential inhibitory properties of curcumin over MMP-9 lead to a therapeutic strategy to reduce the risk of coronary artery involvement during KD.Keywords: MMP-9, coronary artery aneurysm, Kawasaki’s disease, curcumin, AMPK, immune system, NF-κB, MAPK
Procedia PDF Downloads 3041646 Anti-Aging Effects of Two Agricultural Plant Extracts and Their Underlying Mechanism
Authors: Shwu-Ling Peng, Chiung-Man Tsai, Chia-Jui Weng
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Chronic micro-inflammation is a hallmark of many aging-related neurodegenerative and metabolic syndrome-driven diseases. In high glucose (HG) environment, reactive oxygen species (ROS) is generated and the ROS induced inflammation, cytokines secretion, DNA damage, and cell cycle arrest to lead to cellular senescence. Water chestnut shell (WCS) is a plant hull which containing polyphenolic compounds and showed antioxidant and anticancer activities. Orchid, which containing a natural polysaccharide compound, possesses many physiological activities including anti-inflammatory and neuroprotective effects. These agricultural plants might be able to reduce oxidative stress and inflammation. This study was used HG-induced human normal dermal fibroblasts (HG-HNDFs) as an in vitro model to disclose the effects of water extract of Phalaenopsis orchid flower (WEPF) and ethanol extract of water chestnut shell (EEWCS) on the anti-aging and their underlying molecular mechanisms. The toxicity of extracts on human normal dermal fibroblasts (HNDFs) was determined by MTT method. The senescence of cells was assayed by β-galactosidase (SA-β-gal) kit. ROS and nitrate production was analyzed by Intracellular ROS contents and ELISA, respectively. Western blotting was used to detect the proteins in cells. The results showed that the exposure of HNDFs to HG (30 mM) for 72 h were caused cellular senescence and arrested cells at G0/G1 phase. Indeed, the treatment of HG-HNDFs with WEPF (200 μg/ml) and EEWCS (10 μg/ml) significantly released cell cycle arrest and promoted cell proliferation. The G1/S phase transition regulatory proteins such as protein retinoblastoma (pRb), p53, and p16ᴵᴺᴷ⁴ᵃ depressed by WEPF and EEWCS were also observed. Additionally, the treatment of WEPF and EEWCS increased the activity of HO-1 through upregulating Nrf2 as well as decreased the ROS and NO of HG-HNDFs. Therefore, the senescence marker protein-30 (SMP30) in cells was diminished. In conclusion, the WEPF and EEWCS might inhibit HG-induced aging of HNDFs by reducing oxidative stress and free radicals.Keywords: agricultural plant extract, anti-aging, high glucose, Phalaenopsis orchid flower, water chestnut shell
Procedia PDF Downloads 1541645 Synthesis and Characterization of Mixed ligand complexes of Bipyridyl and Glycine with Different Counter Anions as Functional Antioxidant Enzyme Mimics
Authors: Mohamed M. Ibrahim, Gaber A. M. Mersal, Salih Al-Juaid, Samir A. El-Shazly
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A series of mixed ligand complexes, viz., [Cu(BPy)(Gly)X]Y {X = Cl (1), Y = 0; X = 0, Y = ClO4- (2); X = H2O, Y = NO3- (3); X = H2O, Y = CH3COO- (4); and [Cu(BPy)(Gly)-(H2O)]2(SO4) (5) have been synthesized. Their structures and properties were characterized by elemental analysis, thermal analaysis, IR, UV–vis, and ESR spectroscopy, as well as electrochemical measurements including cyclic voltammetry, electrical molar conductivity, and magnetic moment measurements. Complexes 1 and 2 formed slightly distorted square-pyramidal coordination geometries of CuN3OCl and CuN3O2, respectively in which the N,O-donor glycine and N,N-donor bipyridyl bind at the basal plane with chloride ion or water as the axial ligand. Complex 3 shows square planar CuN3O coordination geometry, which exhibits chemically significant hydrogen bonding interactions besides showing coordination polymer formation. The superoxide dismutase and catalase-like activities of all complexes were tested and were found to be promising candidates as durable electron-transfer catalyst being close to the efficiency of the mimicking enzymes displaying either catalase or tyrosinase activity to serve for complete reactive oxygen species (ROS) detoxification, both with respect to superoxide radicals and related peroxides. The DNA binding interaction with super coiled pGEM-T plasmid DNA was investigated by using spectral (absorption and emission) titration and electrochemical techniques. The results revealed that DNA intercalate with complexes 1 and 2 through the groove binding mode. The calculated intrinsic binding constant (Kb) of 1 and 2 were 4.71 and 2.429 × 105 M−1, respectively. Gel electrophoresis study reveals the fact that both complexes cleave super coiled pGEM-T plasmid DNA to nicked and linear forms in the absence of any additives. On the other hand, the interaction of both complexes with DNA, the quasi-reversible CuII/CuI redox couple slightly improves its reversibility with considerable decrease in current intensity. All the experimental results indicate that the bipyridyl mixed copper(II) complex (1) intercalate more effectively into the DNA base pairs.Keywords: enzyme mimics, mixed ligand complexes, X-ray structures, antioxidant, DNA-binding, DNA cleavage
Procedia PDF Downloads 5441644 Physicochemical and Biochemical Characterization of Olea europea Var. Oleaster Oil and Determination of Its Effects on Blood Parameters
Authors: Asma Gherib, Imen Merzougui, Cherifa Henchiri
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This present study has allowed to evaluate the physico chemical characteristics, fatty acid composition and the hypolipidemic effect of Oleaster oil Olea europea var. Oleaster, from the area of El Kala, "Eastern Algeria" on rats "Wistar albinos". The physico chemical characteristics: acidity (0,73%), peroxide value (14, 16 meqO2/kg oil) and iodine value (74,08 g iodine/100 g of oil) are consistent with international standards. The dosage of FA revealed a wealth of oil with UFA (76,7%), mainly composed of 65.43% of MUFA whose major fatty acid is oleic acid (63,57%). The experiment on rats receiving a diet rich in saturated fats and hydrogenated oils revealed that the consumption of Oleaster oil at the dose of 10 g and 20 g for 15 and 30 days improves plasma lipid profile by decreasing the rates of TC, TG, TL, and LDL-C with an increase in the rate of HDL-C serum. The importance of these effects depends on the dose and period of treatment.Keywords: oleaster oil, fatty acid, Olea europea, oleic acid, lipid profile
Procedia PDF Downloads 4881643 Investigating the Expression of NR1/NR2 Receptors in Boys Between 6 to 16 with ADHD Compared to a Healthy Controlled Group
Authors: Sajad Haghshenas
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Emerging evidence from clinical, genetic, and animal model studies suggests that the N-methyl-D-aspartate (NMDA) glutamate receptors (NMDAR) may contribute to the pathophysiology and aetiology of neurological and psychiatric disorders and the patients with impaired NMDR receptors experience psychological symptoms. Therefore, we hypothesised that NMDAR receptors play a key role in the development of attention deficit hyperactivity disorder (ADHD). In this comparative analytical study, we utilized western blotting method to assay the expression levels of NMDA subunits NR1 and NR2 in the blood plasma of 50 male individuals diagnosed with ADHD in comparison to 20 healthy controls. The findings from the western blotting analysis provide support for the hypothesis that individuals with ADHD exhibit significantly lower levels of NR1/2 receptors compared to those without the disorder. Further research is needed to explore the potential causal relationship between reduced NR1/NR2 receptor levels and the development of ADHD.Keywords: expression, glutamate receptors, NR1, NR2, ADHD
Procedia PDF Downloads 731642 Synthesis of Iron-Based Perovskite Type Catalysts from Rust Wastes as a Source of Iron
Authors: M. P. Joshi, F. Deganello, L. F. Liotta, V. La Parola, G. Pantaleo
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For the first time, commercial iron nitrate was replaced by rust wastes, as a source of Iron for the preparation of LaFeO₃ powders by solution combustion synthesis (SCS). A detailed comparison with a reference powder obtained by SCS, starting from a commercial iron nitrate, was also performed. Several techniques such as X-ray diffraction combined with Rietveld refinement, mass plasma atomic emission spectroscopy, nitrogen adsorption measurements, temperature programmed reduction, X-ray photoelectron spectroscopy, Fourier transform analysis and scanning electron microscopy were used for the characterization of the rust wastes as well as of the perovskite powders. The performance of this ecofriendly material was evaluated by testing the activity and selectivity in the propylene oxidation, in order to use it for the benefit of the environment. Characterization and performance results clearly evidenced limitations and peculiarities of this new approach.Keywords: perovskite type catalysts, solution combustion synthesis, X-ray diffraction, rust wastes
Procedia PDF Downloads 3331641 Metal-Semiconductor Transition in Ultra-Thin Titanium Oxynitride Films Deposited by ALD
Authors: Farzan Gity, Lida Ansari, Ian M. Povey, Roger E. Nagle, James C. Greer
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Titanium nitride (TiN) films have been widely used in variety of fields, due to its unique electrical, chemical, physical and mechanical properties, including low electrical resistivity, chemical stability, and high thermal conductivity. In microelectronic devices, thin continuous TiN films are commonly used as diffusion barrier and metal gate material. However, as the film thickness decreases below a few nanometers, electrical properties of the film alter considerably. In this study, the physical and electrical characteristics of 1.5nm to 22nm thin films deposited by Plasma-Enhanced Atomic Layer Deposition (PE-ALD) using Tetrakis(dimethylamino)titanium(IV), (TDMAT) chemistry and Ar/N2 plasma on 80nm SiO2 capped in-situ by 2nm Al2O3 are investigated. ALD technique allows uniformly-thick films at monolayer level in a highly controlled manner. The chemistry incorporates low level of oxygen into the TiN films forming titanium oxynitride (TiON). Thickness of the films is characterized by Transmission Electron Microscopy (TEM) which confirms the uniformity of the films. Surface morphology of the films is investigated by Atomic Force Microscopy (AFM) indicating sub-nanometer surface roughness. Hall measurements are performed to determine the parameters such as carrier mobility, type and concentration, as well as resistivity. The >5nm-thick films exhibit metallic behavior; however, we have observed that thin film resistivity is modulated significantly by film thickness such that there are more than 5 orders of magnitude increment in the sheet resistance at room temperature when comparing 5nm and 1.5nm films. Scattering effects at interfaces and grain boundaries could play a role in thickness-dependent resistivity in addition to quantum confinement effect that could occur at ultra-thin films: based on our measurements the carrier concentration is decreased from 1.5E22 1/cm3 to 5.5E17 1/cm3, while the mobility is increased from < 0.1 cm2/V.s to ~4 cm2/V.s for the 5nm and 1.5nm films, respectively. Also, measurements at different temperatures indicate that the resistivity is relatively constant for the 5nm film, while for the 1.5nm film more than 2 orders of magnitude reduction has been observed over the range of 220K to 400K. The activation energy of the 2.5nm and 1.5nm films is 30meV and 125meV, respectively, indicating that the TiON ultra-thin films are exhibiting semiconducting behaviour attributing this effect to a metal-semiconductor transition. By the same token, the contact is no longer Ohmic for the thinnest film (i.e., 1.5nm-thick film); hence, a modified lift-off process was developed to selectively deposit thicker films allowing us to perform electrical measurements with low contact resistance on the raised contact regions. Our atomic scale simulations based on molecular dynamic-generated amorphous TiON structures with low oxygen content confirm our experimental observations indicating highly n-type thin films.Keywords: activation energy, ALD, metal-semiconductor transition, resistivity, titanium oxynitride, ultra-thin film
Procedia PDF Downloads 2941640 Design of a Dietetic Food: Case of Lebanese Kishk
Authors: Henri El Zakhem, Dona Shalhoub, Elias Atallah, Jessica Koura
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Due to the increase of demand on dietetic food and the need for more types of diet food, the production of dietetic food is increasing and improving. This demand on dietetic food has triggered us to study the market in which we found that Kishk (Lebanese dairy product) diet is not available. Production of a low fat product which is diet Kishk was our concern. A strategy was followed to choose the right idea that will satisfy the need of the market. The whole process was studied and explained thoroughly. The percentage of fat was found to be 32.52 % in regular Kishk and 3.84 % in the diet Kishk produced. The new product has the advantage to be high in protein, low in fat.Keywords: design and industrialization, dietetic, diet Kishk, fat
Procedia PDF Downloads 3751639 Changes in Some Biochemical Parameters and Body Weight of Chicken Exposed to Cadmium
Authors: Khaled Saeed Ali
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This study was conducted with 3 week old domestic chicken to determine the effect of supplementation of cadmium to dietary. 10 mg/kg Cadmium chloride added to maize- sesame cake meal diet for 4 weeks. The additional cadmium to the diet induced a decreasing body weight and changes in biochemical parameters of chicken. Chicken were divided into two groups. The first group was given a diet containing the concentration of 10 mg cadmium /kg daily for a period of 30 days and the second group was given diet without cadmium and used as a control group. The result revealed decrease in the body weight of treated chicken by 12.7 % compared to control group, whose body weight increased. The plasma glucose concentration, creatinine, aspartate aminotranseferase (AST), and alanine aminotransferase (ALT) were increased significantly (P<0.05) in Cd treated chicken in comparison to the control group. Cadmium accumulation was observed in the intestine, kidney, liver and bone. The accumulation of cadmium was markedly higher (3-4 times) in cadmium-treated animals compared to the control.Keywords: cadmium, biochemical parameters, body weight, chicken
Procedia PDF Downloads 4721638 Management of Meskit (Prosopis juliflora) Tree in Oman: The Case of Using Meskit (Prosopis juliflora) Pods for Feeding Omani Sheep
Authors: S. Al-Khalasi, O. Mahgoub, H. Yaakub
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This study evaluated the use of raw or processed Prosopis juliflora (Meskit) pods as a major ingredient in a formulated ration to provide an alternative non-conventional concentrate for livestock feeding in Oman. Dry Meskit pods were reduced to lengths of 0.5- 1.0 cm to ensure thorough mixing into three diets. Meskit pods were subjected to two types of treatments; roasting and soaking. They were roasted at 150оC for 30 minutes using a locally-made roasting device (40 kg barrel container rotated by electric motor and heated by flame gas cooker). Chopped pods were soaked in tap water for 24 hours and dried for 2 days under the sun with frequent turning. The Meskit-pod-based diets (MPBD) were formulated and pelleted from 500 g/kg ground Meskit pods, 240 g/kg wheat bran, 200 g/kg barley grain, 50 g/kg local dried sardines and 10 g/kg of salt. Twenty four 10 months-old intact Omani male lambs with average body weight of 27.3 kg (± 0.5 kg) were used in a feeding trial for 84 days. They were divided (on body weight basis) and allocated to four diet combination groups. These were: Rhodes grass hay (RGH) plus a general ruminant concentrate (GRC); RGH plus raw Meskit pods (RMP) based concentrate; RGH plus roasted Meskit pods (ROMP) based concentrate; RGH plus soaked Meskit pods (SMP) based concentrate Daily feed intakes and bi-weekly body weights were recorded. MPBD had higher contents of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) than the GRC. Animals fed various types of MPBD did not show signs of ill health. There was a significant effect of feeding ROMP on the performance of Omani sheep compared to RMP and SMP. The ROMP fed animals had similar performance to those fed the GRC in terms of feed intake, body weight gain and feed conversion ratio (FCR).This study indicated that roasted Meskit pods based diet may be used instead of the commercial concentrate for feeding Omani sheep without adverse effects on performance. It offers a cheap alternative source of protein and energy for feeding Omani sheep. Also, it might help in solving the spread impact of Meskit trees, maintain the ecosystem and helping in preserving the local tree species.Keywords: growth, Meskit, Omani sheep, Prosopis juliflora
Procedia PDF Downloads 4761637 Impact of Different Rearing Diets on the Performance of Adult Mealworms Tenebrio molitor
Authors: Caroline Provost, Francois Dumont
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Production of insects for human and animal consumption is an increasingly important activity in Canada. Protein production is more efficient and less harmful to the environment using insect rearing compared to the impact of traditional livestock, poultry and fish farms. Insects are rich in essential amino acids, essential fatty acids and trace elements. Thus, insect-based products could be used as a food supplement for livestock and domestic animals and may even find their way into the diets of high performing athletes or fine dining. Nevertheless, several parameters remain to be determined to ensure efficient and profitable production that meet the potential of these sectors. This project proposes to improve the production processes, rearing diets and processing methods for three species with valuable gastronomic and nutritional potential: the common mealworms (Tenebrio molitor), the small mealworm (Alphitobius diaperinus), and the giant mealworm (Zophobas morio). The general objective of the project is to acquire specific knowledge for mass rearing of insects dedicated to animal and human consumption in order to respond to current market opportunities and meet a growing demand for these products. Mass rearing of the three species of mealworm was produced to provide the individuals needed for the experiments. Mealworms eat flour from different cereals (e.g. wheat, barley, buckwheat). These cereals vary in their composition (protein, carbohydrates, fiber, vitamins, antioxidant, etc.), but also in their purchase cost. Seven different diets were compared to optimize the yield of the rearing. Diets were composed of cereal flour (e.g. wheat, barley) and were either mixed or left alone. Female fecundity, larvae mortality and growing curves were observed. Some flour diets have positive effects on female fecundity and larvae performance while each mealworm was found to have specific diet requirements. Trade-offs between mealworm performance and costs need to be considered. Experiments on the effect of flour composition on several parameters related to performance and nutritional and gastronomic value led to the identification of a more appropriate diet for each mealworm.Keywords: mass rearing, mealworm, human consumption, diet
Procedia PDF Downloads 1471636 On the Other Side of Shining Mercury: In Silico Prediction of Cold Stabilizing Mutations in Serine Endopeptidase from Bacillus lentus
Authors: Debamitra Chakravorty, Pratap K. Parida
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Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design
Procedia PDF Downloads 1401635 A Fluorescent Polymeric Boron Sensor
Authors: Soner Cubuk, Mirgul Kosif, M. Vezir Kahraman, Ece Kok Yetimoglu
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Boron is an essential trace element for the completion of the life circle for organisms. Suitable methods for the determination of boron have been proposed, including acid - base titrimetric, inductively coupled plasma emission spectroscopy flame atomic absorption and spectrophotometric. However, the above methods have some disadvantages such as long analysis times, requirement of corrosive media such as concentrated sulphuric acid and multi-step sample preparation requirements and time-consuming procedures. In this study, a selective and reusable fluorescent sensor for boron based on glycosyloxyethyl methacrylate was prepared by photopolymerization. The response characteristics such as response time, pH, linear range, limit of detection were systematically investigated. The excitation/emission maxima of the membrane were at 378/423 nm, respectively. The approximate response time was measured as 50 sec. In addition, sensor had a very low limit of detection which was 0.3 ppb. The sensor was successfully used for the determination of boron in water samples with satisfactory results.Keywords: boron, fluorescence, photopolymerization, polymeric sensor
Procedia PDF Downloads 2831634 Fine Characterization of Glucose Modified Human Serum Albumin by Different Biophysical and Biochemical Techniques at a Range
Authors: Neelofar, Khursheed Alam, Jamal Ahmad
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Protein modification in diabetes mellitus may lead to early glycation products (EGPs) or amadori product as well as advanced glycation end products (AGEs). Early glycation involves the reaction of glucose with N-terminal and lysyl side chain amino groups to form Schiff’s base which undergoes rearrangements to form more stable early glycation product known as Amadori product. After Amadori, the reactions become more complicated leading to the formation of advanced glycation end products (AGEs) that interact with various AGE receptors, thereby playing an important role in the long-term complications of diabetes. Millard reaction or nonenzymatic glycation reaction accelerate in diabetes due to hyperglycation and alter serum protein’s structure, their normal functions that lead micro and macro vascular complications in diabetic patients. In this study, Human Serum Albumin (HSA) with a constant concentration was incubated with different concentrations of glucose at 370C for a week. At 4th day, Amadori product was formed that was confirmed by colorimetric method NBT assay and TBA assay which both are authenticate early glycation product. Conformational changes in native as well as all samples of Amadori albumin with different concentrations of glucose were investigated by various biophysical and biochemical techniques. Main biophysical techniques hyperchromacity, quenching of fluorescence intensity, FTIR, CD and SDS-PAGE were used. Further conformational changes were observed by biochemical assays mainly HMF formation, fructoseamine, reduction of fructoseamine with NaBH4, carbonyl content estimation, lysine and arginine residues estimation, ANS binding property and thiol group estimation. This study find structural and biochemical changes in Amadori modified HSA with normal to hyperchronic range of glucose with respect to native HSA. When glucose concentration was increased from normal to chronic range biochemical and structural changes also increased. Highest alteration in secondary and tertiary structure and conformation in glycated HSA was observed at the hyperchronic concentration (75mM) of glucose. Although it has been found that Amadori modified proteins is also involved in secondary complications of diabetes as AGEs but very few studies have been done to analyze the conformational changes in Amadori modified proteins due to early glycation. Most of the studies were found on the structural changes in Amadori protein at a particular glucose concentration but no study was found to compare the biophysical and biochemical changes in HSA due to early glycation with a range of glucose concentration at a constant incubation time. So this study provide the information about the biochemical and biophysical changes occur in Amadori modified albumin at a range of glucose normal to chronic in diabetes. Although many implicates currently in use i.e. glycaemic control, insulin treatment and other chemical therapies that can control many aspects of diabetes. However, even with intensive use of current antidiabetic agents more than 50 % of diabetic patient’s type 2 suffers poor glycaemic control and 18 % develop serious complications within six years of diagnosis. Experimental evidence related to diabetes suggests that preventing the nonenzymatic glycation of relevant proteins or blocking their biological effects might beneficially influence the evolution of vascular complications in diabetic patients or quantization of amadori adduct of HSA by authentic antibodies against HSA-EGPs can be used as marker for early detection of the initiation/progression of secondary complications of diabetes. So this research work may be helpful for the same.Keywords: diabetes mellitus, glycation, albumin, amadori, biophysical and biochemical techniques
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