Search results for: protein extraction

Authors: Zainab S. Ahmed, Mohammed S. Ali, Nadia A. Elshiekh, Sami Adam Ibrahim, Ghada M. El-Tayeb, Ahmed H. Elsadig, Rihab A. Omer, Sofia B. Mohamed

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Maple syrup urine (MSUD) is an autosomal recessive disease that causes a deficiency in the enzyme branched-chain alpha-keto acid (BCKA) dehydrogenase. The development of disease has been associated with SNPs in the DBT gene. Despite that, the computational analysis of SNPs in coding and noncoding and their functional impacts on protein level still remains unknown. Hence, in this study, we carried out a comprehensive in silico analysis of missense that was predicted to have a harmful influence on DBT structure and function. In this study, eight different in silico prediction algorithms; SIFT, PROVEAN, MutPred, SNP&GO, PhD-SNP, PANTHER, I-Mutant 2.0 and MUpo were used for screening nsSNPs in DBT including. Additionally, to understand the effect of mutations in the strength of the interactions that bind protein together the ELASPIC servers were used. Finally, the 3D structure of DBT was formed using Mutation3D and Chimera servers respectively. Our result showed that a total of 15 nsSNPs confirmed by 4 software (R301C, R376H, W84R, S268F, W84C, F276C, H452R, R178H, I355T, V191G, M444T, T174A, I200T, R113H, and R178C) were found damaging and can lead to a shift in DBT gene structure. Moreover, we found 7 nsSNPs located on the 2-oxoacid_dh catalytic domain, 5 nsSNPs on the E_3 binding domain and 3 nsSNPs on the Biotin Domain. So these nsSNPs may alter the putative structure of DBT’s domain. Furthermore, we detected all these nsSNPs are on the core residues of the protein and have the ability to change the stability of the protein. Additionally, we found W84R, S268F, and M444T have high significance, and they affected Leucine, Isoleucine, and Valine, which reduces or disrupt the function of BCKD complex, E2-subunit which the DBT gene encodes. In conclusion, based on our extensive in-silico analysis, we report 15 nsSNPs that have possible association with protein deteriorating and disease-causing abilities. These candidate SNPs can aid in future studies on Maple Syrup Urine Disease type II base in the genetic level.

Keywords: DBT gene, ELASPIC, in silico analysis, UCSF chimer

Procedia PDF Downloads 187

Authors: J. J. Lee, S. R. Sung, E. J. Yum, S. C. Kim, B. H. Hyun, M. Her, H. S. Lee

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Canine brucellosis is a critical problem in dogs leading to reproductive diseases which are mainly caused by Brucella canis. There are, nonetheless, not clear symptoms so that it may go unnoticed in most of the cases. Serodiagnosis for canine brucellosis has not been confirmed. Moreover, it has substantial difficulties due to broad cross-reactivity between the rough cell wall antigens of B. canis and heterospecific antibodies present in normal, uninfected dogs. Thus, this study was conducted to characterize the immunogenic proteins in cytoplasmic antigen (CPAg) of B. canis, which defined the antigenic sensitivity of the humoral antibody responses to B. canis-infected dogs. In analysis of B. canis CPAg, first, we extracted and purified the cytoplasmic proteins from cultured B. canis by hot-saline inactivation, ultrafiltration, sonication, and ultracentrifugation step by step according to the sonicated antigen extract method. For characterization of this antigen, we checked the sort and range of each protein on SDS-PAGE and verified the immunogenic proteins leading to reaction with antisera of B. canis-infected dogs. Selected immunodominant proteins were identified using MALDI-MS/MS. As a result, in an immunoproteomic assay, several polypeptides in CPAg on one or two-dimensional electrophoresis (DE) were specifically reacted to antisera from B. canis-infected dogs but not from non-infected dogs. The polypeptides with approximate 150, 80, 60, 52, 33, 26, 17, 15, 13, 11 kDa on 1-DE were dominantly recognized by antisera from B. canis-infected dogs. In the immunoblot profiles on 2-DE, ten immunodominant proteins in CPAg were detected with antisera of infected dogs between pI 3.5-6.5 at approximate 35 to 10 KDa, without any nonspecific reaction with sera in non-infected dogs. Ten immunodominant proteins identified by MALDI-MS/MS were identified as superoxide dismutase, bacteroferritin, amino acid ABC transporter substrate-binding protein, extracellular solute-binding protein family3, transaldolase, 26kDa periplasmic immunogenic protein, Rhizopine-binding protein, enoyl-CoA hydratase, arginase and type1 glyceraldehyde-3-phosphate dehydrogenase. Most of these proteins were determined by their cytoplasmic or periplasmic localization with metabolism and transporter functions. Consequently, this study discovered and identified the prominent immunogenic proteins in B. canis CPAg, highlighting that those antigenic proteins may accomplish a specific serodiagnosis for canine brucellosis. Furthermore, we will evaluate those immunodominant proteins for applying to the advanced diagnostic methods with high specificity and accuracy.

Keywords: Brucella canis, Canine brucellosis, cytoplasmic antigen, immunogenic proteins

Procedia PDF Downloads 129

Authors: Seok Hong Min, Tae Kwon Ha

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With the increasing interest on Ti alloys, the extraction process of Ti from its typical ore, TiO₂, has long been and will be important issue. As an intermediate product for the production of pigment or titanium metal sponge, tetrachloride (TiCl₄) is produced by fluidized bed using high TiO₂ feedstock. The purity of TiCl₄ after chlorination is subjected to the quality of the titanium feedstock. Since the impurities in the TiCl₄ product are reported to final products, the purification process of the crude TiCl₄ is required. The purification process includes fractional distillation and chemical treatment, which depends on the nature of the impurities present and the required quality of the final product. In this study, thermodynamic analysis on the impurity effect in the chlorination process, which is the first step of extraction of Ti from TiO₂, has been conducted. All thermodynamic calculations were performed using the FactSage thermodynamical software.

Keywords: rutile, titanium, chlorination process, impurities, thermodynamic calculation, FactSage

Procedia PDF Downloads 290

Authors: Nadjiba Mezıou-Cheboutı, Amel Merabet, Yahia Cheboutı, Nassima Behidj, Fatima-Zohra Bıssaad et Salahedine Doumandjı

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Among the Anacardiaceae, the fruit (Pistacia vera L.) is the only species that produces edible fruits. The introduction of real pistachio was made in the early sixties by an FAO program in Algeria in several regions in the northern part of Algeria: Tlemcen, Sidi Bel Abbes, Batna, Bouira M'sila . Chemical analyzes of seeds pistachios were made on seeds from an orchard that localizes to Bechloul (Bouira) located in bioclimatic sub-humid temperate winter floor. Analyzes reveal dry matter content of 3.60 ± 0.45%, the water rate is 7.21 ± 0.36%. However, the fat content is 46.00 ± 0.90%, in average blood sugar, it is 4.02 ± 0.47%, the protein reached 29.88 ± 0.76%. Given the very interesting that high-fat food nutritional values, culture pistachio must be considered for its extension in Algeria.

Keywords: pistachio, dry matter, fat, sugar, protein

Procedia PDF Downloads 337

Authors: Nurlan Akhmetov, Abilmansur Yeshmuratov, Aliya Kurbanova, Gulnar Sugurbekova, Murat Baisariyev

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Extraction of the vanadium and nickel compounds is complex due to the high stability of porphyrin, nickel is catalytic poison which deactivates catalysis during the catalytic cracking of the oil, while vanadyl is abrasive and valuable metal. Thus, high concentration of the Ni and V in the crude oil makes their removal relevant. Two methods of the demetallization of crude oil were tested, therefore, the present research is conducted for comparative analysis of the deasphalting with organic solvents (cyclohexane, carbon tetrachloride, chloroform) and electrochemical method. Percentage of Ni extraction reached maximum of approximately 55% by using the electrochemical method in electrolysis cell, which was developed for this research and consists of three sections: oil and protonating agent (EtOH) solution between two conducting membranes which divides it from two capsules of 10% sulfuric acid and two graphite electrodes which cover all three parts in electrical circuit. Ions of metals pass through membranes and remain in acid solutions. The best result was obtained in 60 minutes with ethanol to oil ratio 25% to 75% respectively, current fits in to the range from 0.3A to 0.4A, voltage changed from 12.8V to 17.3V. Maximum efficiency of deasphalting, with cyclohexane as the solvent, in Soxhlet extractor was 66.4% for Ni and 51.2% for V. Thus, applying the voltammetry, ICP MS (Inductively coupled plasma mass spectrometry) and AAS (atomic absorption spectroscopy), these mentioned types of metal extraction methods were compared in this paper.

Keywords: electrochemistry, deasphalting of crude oil, demetallization of crude oil, petrolium engineering

Procedia PDF Downloads 216

Authors: Nitin Shravan, Sudarsun Santhiappan, B. Sivaselvan

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Subsection identification, in the context of Electronic Health Records (EHRs), is identifying the important sections for down-stream tasks like auto-coding. In this work, we classify the text present in EHRs according to their information, using machine learning and deep learning techniques. We initially describe briefly about the problem and formulate it as a text classification problem. Then, we discuss upon the methods from the literature. We try two approaches - traditional feature extraction based machine learning methods and deep learning methods. Through experiments on a private dataset, we establish that the deep learning methods perform better than the feature extraction based Machine Learning Models.

Keywords: deep learning, machine learning, semantic clinical classification, subsection identification, text classification

Procedia PDF Downloads 195

Authors: Issam Ben Jami

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Information retrieval in digital libraries is very important because most famous historical documents occupy a significant value. The word spotting in historical documents is a very difficult notion, because automatic recognition of such documents is naturally cursive, it represents a wide variability in the level scale and translation words in the same documents. We first present a system for the automatic recognition, based on the extraction of interest points words from the image model. The extraction phase of the key points is chosen from the representation of the image as a synthetic description of the shape recognition in a multidimensional space. As a result, we use advanced methods that can find and describe interesting points invariant to scale, rotation and lighting which are linked to local configurations of pixels. We test this approach on documents of the 15th century. Our experiments give important results.

Keywords: feature matching, historical documents, pattern recognition, word spotting

Procedia PDF Downloads 261

Authors: Amr Mohamed Bakry Ibrahim, Yingzhou Ni, Hao Cheng, Li Liang

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Tuna oil (omega-3 oil) has become increasingly popular in the last ten years, because it is considered one of the treasures of food which has many beneficial health effects for the humans. Nevertheless, the susceptibility of omega-3 oils to oxidative deterioration, resulting in the formation of oxidation products, in addition to organoleptic problems including “fishy” flavors, have presented obstacles to the more widespread use of tuna oils in the food industry. This study sought to evaluate the potential impact of Mentha piperita oil on physicochemical characteristics and oxidative stability of tuna oil microcapsules formed by spray drying using the partial substitution to whey protein isolate by carboxymethyl cellulose and pullulan. The emulsions before the drying process were characterized regarding size and ζ-potential, viscosity, surface tension. Confocal laser scanning microscopy showed that all emulsions were sphericity and homogeneous distribution without any visible particle aggregation. The microcapsules obtained after spray drying were characterized regarding microencapsulation efficiency, water activity, color, bulk density, flowability, scanning surface morphology and oxidative stability. The microcapsules were spherical shape had low water activity (0.11-0.23 aw). The microcapsules containing both tuna oil and Mentha piperita oil were smaller than others and addition of pullulan into wall materials improved the morphology of microcapsules. Microencapsulation efficiency of powdered oil ranged from 90% to 94%. Using Mentha piperita oil in the process of microencapsulation tuna oil enhanced the oxidative stability using whey protein isolate only or with carboxymethyl cellulose or pullulan as wall materials, resulting in improved storage stability and mask fishy odor. Therefore, it is foreseen using tuna-Mentha piperita oil mixture microcapsules in the applications of the food industries.

Keywords: Mentha piperita oil, microcapsule, tuna oil, whey protein isolate

Procedia PDF Downloads 336

Authors: Jean Paul Hategekimana, Bampire Claudine, Niyonsenga Nadia, Irakoze Josiane

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Peas, soybeans and rice are crops which are grown in Rwanda and are available in rural and urban local markets and they give contribution in reduction of health problems especially in fighting malnutrition and food insecurity in Rwanda. Several research activities have been conducted on how cereals flour can be mixed with legumes flour for developing baked products which are rich in protein, fiber, minerals as they are found in legumes. However, such activity was not yet well studied in Rwanda. The aim of the present study was to develop bread and biscuit products from peas, soybeans and rice as functional ingredients combined with wheat flour and then analyze the nutritional content and consumer acceptability of new developed products. The malnutrition problem can be reduced by producing bread and biscuits which are rich in protein and are very accessible for every individual. The processing of bread and biscuit were made by taking peas flour, soybeans flour and rice flour mixed with wheat flour and other ingredients then a dough was made followed by baking. For bread, two kind of products were processed, for each product one control and three experimental samples in different three ratios of peas and rice were prepared. These ratios were 95:5, 90:10 and 80:20 for bread from peas and 85:5:10, 80:10:10 and 70:10:20 for bread from peas and rice. For biscuit, two kind of products were also processed, for each product one control sample and three experimental samples in three different ratios were prepared. These ratios are 90:5:5,80:10:10 and 70:10:20 for biscuit from peas and rice and 90:5:5,80:10:10 and 70:10:20 for biscuit from soybean and rice. All samples including the control sample were analyzed for the consumer acceptability (sensory attributes) and nutritional composition. For sensory analysis, bread from of peas and rice flour with wheat flour at ratio 85:5:10 and bread from peas only as functional ingredient with wheat flour at ratio 95:5 and biscuits made from a of soybeans and rice at a ratio 90:5:5 and biscuit made from peas and rice at ratio 90:5:5 were most acceptable compared to control sample and other samples in different ratio. The moisture, protein, fat, fiber and minerals (Sodium and iron.) content were analyzed where bread from peas in all ratios was found to be rich in protein and fiber compare to control sample and biscuit from soybean and rice in all ratios was found to be rich in protein and fiber compare to control sample.

Keywords: bakery products, peas and rice flour, wheat flour, sensory evaluation, proximate composition

Procedia PDF Downloads 47

Authors: Arash Rafiei, Carroll Moore

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Leaching process as the first stage of hydrometallurgy is a multidisciplinary system including material properties, chemistry, reactor design, mechanics and fluid dynamics. Therefore, doing leaching system optimization by pure scientific methods need lots of times and expenses. In this work, a mixture of two titanium ores and one titanium slag are used for extracting titanium for leaching stage of TiO2 pigment production procedure. Optimum titanium extraction can be obtained from following strategies: i) Maximizing titanium extraction without selective digestion; and ii) Optimizing selective titanium extraction by balancing between maximum titanium extraction and minimum impurity digestion. The main difference between two strategies is due to process optimization framework. For the first strategy, the most important stage of production process is concerned as the main stage and rest of stages would be adopted with respect to the main stage. The second strategy optimizes performance of more than one stage at once. The second strategy has more technical complexity compared to the first one but it brings more economical and technical advantages for the leaching system. Obviously, each strategy has its own optimum operational zone that is not as same as the other one and the best operational zone is chosen due to complexity, economical and practical aspects of the leaching system. Experimental design has been carried out by using Taguchi method. The most important advantages of this methodology are involving different technical aspects of leaching process; minimizing the number of needed experiments as well as time and expense; and concerning the role of parameter interactions due to principles of multifactor-at-time optimization. Leaching tests have been done at batch scale on lab with appropriate control on temperature. The leaching tank geometry has been concerned as an important factor to provide comparable agitation conditions. Data analysis has been done by using reactor design and mass balancing principles. Finally, optimum zone for operational parameters are determined for each leaching strategy and discussed due to their economical and practical aspects.

Keywords: titanium leaching, optimization, experimental design, performance analysis

Procedia PDF Downloads 355

Authors: U. Ravi Babu, Mohd Mastan

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The present paper proposes a novel approach for handwritten digit recognition system. The present paper extract digit image features based on distance measure and derives an algorithm to classify the digit images. The distance measure can be performing on the thinned image. Thinning is the one of the preprocessing technique in image processing. The present paper mainly concentrated on an extraction of features from digit image for effective recognition of the numeral. To find the effectiveness of the proposed method tested on MNIST database, CENPARMI, CEDAR, and newly collected data. The proposed method is implemented on more than one lakh digit images and it gets good comparative recognition results. The percentage of the recognition is achieved about 97.32%.

Keywords: handwritten digit recognition, distance measure, MNIST database, image features

Procedia PDF Downloads 445

Authors: Mario Kubek, Herwig Unger

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Due to the large amount of information in the World Wide Web (WWW, web) and the lengthy and usually linearly ordered result lists of web search engines that do not indicate semantic relationships between their entries, the search for topically similar and related documents can become a tedious task. Especially, the process of formulating queries with proper terms representing specific information needs requires much effort from the user. This problem gets even bigger when the user's knowledge on a subject and its technical terms is not sufficient enough to do so. This article presents the new and interactive search application DocAnalyser that addresses this problem by enabling users to find similar and related web documents based on automatic query formulation and state-of-the-art search word extraction. Additionally, this tool can be used to track topics across semantically connected web documents

Keywords: DocAnalyser, interactive web search, search word extraction, query formulation, source topic detection, topic tracking

Procedia PDF Downloads 382

Authors: Nadjiba Meziou-Chebouti, Amel Merabet, Yahia Chebouti, Nassima Behidj, Salahedine Doumandji

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Among the Anacardiaceae, the fruit (Pistacia vera L.) is the only species that produces edible fruits. The introduction of real pistachio was made in the early sixties by an FAO program in Algeria in several regions in the northern part of Algeria: Tlemcen, Sidi Bel Abbes, Batna, Bouira M'sila. Chemical analyzes of seeds pistachios were made on seeds from an orchard that localizes to Bechloul (Bouira) located in bioclimatic sub-humid temperate winter floor. Analyzes reveal dry matter content of 3.60±0.45%, the water rate is 7.21±0.36%. However, the fat content is 46.00±0.90%, in average blood sugar, it is 4.02±0.47%, the protein reached 29.88±0.76%. Given the very interesting that high-fat food nutritional values, culture pistachio must be considered for its extension in Algeria.

Keywords: pistachio, dry matter, fat, sugar, protein

Procedia PDF Downloads 353

Authors: S. Zouainia, R. Djebar, H. Berrebah, A. Sayeb

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This work focused on the study of physiological and biochemical changes observed in tadpoles exposed to fungicide Rana saharica Artea 330ec recently introduced in Algeria. For this, we tested the effect of xenobiotics on growth and development of tadpoles; among the studied parameters: total protein, glutathione and respiratory activity. The study of physiological parameters showed that the tadpoles change perfectly in the absence of toxic and in favorable conditions (pH, temperature). Our results showed an increased rate of protein and GSH in the presence of the fungicide Artea 330ec. The latter causes uninhibited very highly significant respiratory activity of tadpoles treated. The presence of xenobiotics in the breeding tadpoles water causes disturbances in behavior and food metabolism.

Keywords: amphibians, fungicides, pesticides, pollution

Procedia PDF Downloads 390

Authors: Simona Dostalova, Dita Munzova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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The boom of nanomedicine in recent years has led to the development of numerous new nanomaterials that can be used as nanocarriers in the drug delivery. These nanocarriers can either be synthetic or natural-based. The disadvantage of many synthetic nanocarriers is their toxicity in patient’s body. Protein cages that can naturally be found in human body do not exhibit such disadvantage. However, the release of cargo from some protein cages in target cells can be problematic. As a special type of protein cages can serve the capsid of many viruses, including phage. Phages infect bacterial cells; therefore they are not harmful to human cells. The targeting of phage particles to cancer cells can be solved by producing of empty phage capsids during which the targeting moieties (e.g. peptides) can be cloned into genes of phage capsid to decorate its surface. Moreover, the produced capsids do not contain viral nucleic acid and are therefore not infectious to beneficial bacteria in the patient’s body. The protein cage composed of viral capsid is larger than other frequently used apoferritin cage but its size is still small enough to benefit from passive targeting by Enhanced Permeability and Retention effect. In this work, bacteriophage λ was used, both whole and its empty capsid for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin, etoposide and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2 % maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000 g and 4 °C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 µg/ml). Free molecules of drugs were removed by dialysis. The encapsulation was verified using spectrophotometric and electrochemical methods. The amount of encapsulated drug linearly increased with the amount of applied drug (determination coefficient R2=0.8013). 76% of applied drug was encapsulated in phage λ particles (concentration of 10 µg/ml), even with the highest applied concentration of drugs, 200 µg/ml. Only 1% of encapsulated drug was detected in phage DNA. Similar results were obtained with encapsulation in phage empty capsid. Therefore, it can be concluded that the encapsulation of drugs into phage particles is efficient and mostly occurs by interaction of drugs with protein capsid.

Keywords: cytostatics, drug delivery, nanocarriers, phage capsid

Procedia PDF Downloads 475

Authors: Versha Sharma

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Juvenile hormone analogue, fenoxycarb and ecdysterone, when applied at varying concentrations in the adult females of Dysdercus similis, in situ histochemical observations of treated ovarian and adipose tissues during the first gonotrophic cycle elicited drastic histomorphological changes in both tissues. The action and effect of both JHa and ecdysterone on ovarian development, vitellogenesis, the activity of follicular epithelium, chorion formation all were monitored in detail. SDS-PAGE electrophoretic analysis showed drastic downregulation on the protein profile of differently treated tissue samples. After exogenous JHa supply, resorption of the developing oocytes was also often noticed. Gradational decline and disappearance of different protein bands in treated both ovarian and adipose tissues noticed could be due to the depletion of specific metabolites essential for oocyte development and maturation. Natural products support both crop production and the environment that being effective in pest control, less toxic to non-target organisms and at the same time biodegradable. Hence, these could be utilized as an attractive alternative to the synthetic chemical insecticides for at least cotton bug pest management. Increasing IGR dosages is found to elicit both qualitative and quantitative depletion of protein metabolites and drastic histochemical changes in the gonads of the treated forms brought forth the production of a large number of immature mal-formed oocytes. Findings in greater detail could be discussed.

Keywords: juvenile hormone, ecdysone, P. picta, Dysdercus similis

Procedia PDF Downloads 236

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat

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We hypothesized that rexinoids such as bexarotene, a selective retinoid X receptor α (RXRα) agonist, may be beneficial for preventing mortality due to inflammation associated with increased expression/activity of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide (LPS). Therefore, we investigated effects of bexarotene on the changes in circulating protein levels of iNOS (an index for systemic iNOS expression), myeloperoxidase (MPO) (an index for systemic inflammation), and lactate dehydrogenase (LDH) (an index for systemic tissue injury) in LPS-induced systemic inflammation model resulting in septic shock in rats. Rats were injected with saline (4 ml/kg; i.p.), LPS (10 mg/kg; i.p.), dimethylsulphoxide (4 ml/kg, 0.1%; s.c.) at time 0. Mean arterial blood pressure and heart rate were measured using a tail-cuff device. Bexarotene (0.03, 0.1, 0.3, and 1 mg/kg; s.c.) was administered to separate groups of rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and blood was collected for measurement of serum iNOS, MPO, and LDH protein levels. Blood pressure decreased by 31 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. Bexarotene at 0.3 and 1 mg/kg doses caused 20% mortality 4 h after LPS injection. In the LPS-treated rats, serum iNOS, MPO, and LDH protein levels were increased. Bexarotene only at 0.1 mg/kg dose prevented the LPS-induced hypotension and increased in iNOS, MPO, and LDH protein levels. These data are consistent with the view that a decrease in systemic iNOS levels contributes to the beneficial effect of bexarotene to prevent the hypotension associated with inflammation and tissue injury during rat endotoxemia. [This work was financially supported by The Scientific and Technological Research Council of Turkey (SBAG-109S121)].

Keywords: bexarotene, inflammation, iNOS, lipopolisaccharide, RXRa

Procedia PDF Downloads 297

Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

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The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

Procedia PDF Downloads 148

Authors: Rafia Asghar, Abdul Hafeez

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The current research was related with natural dyes’ extraction from the powder of Neem (Azadirachta indica) bark and studied characterization of this dye under microwave radiation’s influence. Both cotton fabric and dyeing powder were exposed to microwave rays for different time intervals (2minutes, 4 minutes, 6 minutes, 8 minutes and 10 minutes) using conventional oven. Aqueous, 60% Methanol and Ethyl Acetate solubilized extracts obtained from Neem (Azadirachta indica) bark were also exposed to different time intervals (2minutes, 4 minutes, 6 minutes, 8 minutes and 10 minutes) of microwave rays exposure. Pre, meta and post mordanting with Alum (2%, 4%, 6%, 8%, and 10%) was done to improve color strength of the extracted dye. Exposure of Neem (Azadirachta indica) bark extract and cotton to microwave rays enhanced the extraction process and dyeing process by reducing extraction time, dyeing time and dyeing temperature. Microwave rays treatment had a very strong influence on color fastness and color strength properties of cotton that was dyes using Neem (Azadirachta indica) bark for 30 minutes and dyeing cotton with that Neem bark extract for 75 minutes at 30°C. Among pre, meta and post mordanting, results indicated that 5% concentration of Alum in meta mordanting exhibited maximum color strength.

Keywords: dyes, natural dyeing, ecofriendly dyes, microwave treatment

Procedia PDF Downloads 677

Authors: Jana Chakrabarti, Madhushrita Das, Ankhi Haldar, Roshni Chatterjee, Tanmoy Dey, Pubali Dhar

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High incidences of Protein Energy Malnutrition as a consequence of low protein intake are quite prevalent among the children in developing countries. Thus prevention of under-nutrition has emerged as a critical challenge to India’s developmental Planners in recent times. Increase in population over the last decade has led to greater pressure on the existing animal protein sources. But these resources are currently declining due to persistent drought, diseases, natural disasters, high-cost of feed, and low productivity of local breeds and this decline in productivity is most evident in some developing countries. So the need of the hour is to search for efficient utilization of unconventional low-cost animal protein resources. Molluscs, as a group is regarded as under-exploited source of health-benefit molecules. Bivalve is the second largest class of phylum Mollusca. Annual harvests of bivalves for human consumption represent about 5% by weight of the total world harvest of aquatic resources. The freshwater mussel Lamellidens marginalis is widely distributed in ponds and large bodies of perennial waters in the Indian sub-continent and well accepted as food all over India. Moreover, ethno-medicinal uses of the flesh of Lamellidens among the rural people to treat hypertension have been documented. Present investigation thus attempts to evaluate the potential of Lamellidens marginalis as functional food. Mussels were collected from freshwater ponds and brought to the laboratory two days before experimentation for acclimatization in laboratory conditions. Shells were removed and fleshes were preserved at- 20oC until analysis. Tissue homogenate was prepared for proximate studies. Fatty acids and amino acids composition were analyzed. Vitamins, Minerals and Heavy metal contents were also studied. Mussel Protein hydrolysate was prepared using Alcalase 2.4 L and degree of hydrolysis was evaluated to analyze its Functional properties. Ferric Reducing Antioxidant Power (FRAP) and DPPH Antioxidant assays were performed. Anti-hypertensive property was evaluated by measuring Angiotensin Converting Enzyme (ACE) inhibition assay. Proximate analysis indicates that mussel meat contains moderate amount of protein (8.30±0.67%), carbohydrate (8.01±0.38%) and reducing sugar (4.75±0.07%), but less amount of fat (1.02±0.20%). Moisture content is quite high but ash content is very low. Phospholipid content is significantly high (19.43 %). Lipid constitutes, substantial amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which have proven prophylactic values. Trace elements are found present in substantial amount. Comparative study of proximate nutrients between Labeo rohita, Lamellidens and cow’s milk indicates that mussel meat can be used as complementary food source. Functionality analyses of protein hydrolysate show increase in Fat absorption, Emulsification, Foaming capacity and Protein solubility. Progressive anti-oxidant and anti-hypertensive properties have also been documented. Lamellidens marginalis can thus be regarded as a functional food source as this may combine effectively with other food components for providing essential elements to the body. Moreover, mussel protein hydrolysate provides opportunities for utilizing it in various food formulations and pharmaceuticals. The observations presented herein should be viewed as a prelude to what future holds.

Keywords: functional food, functional properties, Lamellidens marginalis, protein hydrolysate

Procedia PDF Downloads 407

Authors: Daniela Rivera-Tobar Mario Perez-Won, Roberto Lemus-Mondaca, Gipsy Tabilo-Munizaga

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Different dietary trends worldwide seek to consume foods with anti-inflammatory properties, rich in antioxidants, proteins, and unsaturated fatty acids that lead to better metabolic, intestinal, mental, and cardiac health. In this sense, food matrices with high protein content based on macro and microalgae are an excellent alternative to meet the new needs of consumers. An emerging and environmentally friendly technology for producing protein matrices is ultrasonic agglomeration. It consists of the formation of permanent bonds between particles, improving the agglomeration of the matrix compared to conventionally agglomerated products (compression). Among the advantages of this process are the reduction of nutrient loss and the avoidance of binding agents. The objective of this research was to optimize the ultrasonic agglomeration process in matrices composed of Spirulina (Arthrospira platensis) powder and Cochayuyo (Durvillae Antartica) flour, by means of the response variable (Young's modulus) and the independent variables were the process conditions (percentage of ultrasonic amplitude: 70, 80 and 90; ultrasonic agglomeration times and cycles: 20, 25 and 30 seconds, and 3, 4 and 5). It was evaluated using a central composite design and analyzed using response surface methodology. In addition, the effects of agglomeration on thermophysical and microstructural properties were evaluated. It was determined that ultrasonic compression with 80 and 90% amplitude caused conformational changes according to Fourier infrared spectroscopy (FTIR) analysis, the best condition with respect to observed microstructure images (SEM) and differential scanning calorimetry (DSC) analysis, was the condition of 90% amplitude 25 and 30 seconds with 3 and 4 cycles of ultrasound. In conclusion, the agglomerated matrices present good macro and microstructural properties which would allow the design of food systems with better nutritional and functional properties.

Keywords: ultrasonic agglomeration, physical properties of food, protein matrices, macro and microalgae

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Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: Pratima Kumari, Sukha Ranjan Samadder

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This research aims to contribute to the sustainable production of graphene materials by exploring the extraction of graphite from spent primary and secondary batteries. The increasing demand for graphene materials, a versatile and high-performance material, necessitates environmentally friendly methods for its synthesis. The process involves a well-planned methodology, beginning with the gathering and categorization of batteries, followed by the disassembly and careful removal of graphite from anode structures. The use of environmentally friendly solvents and mechanical techniques ensures an efficient and eco-friendly extraction of graphite. Advanced approaches such as the modified Hummers' method and chemical reduction process are utilized for the synthesis of graphene materials, with a focus on optimizing parameters. Various analytical techniques such as Fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, thermogravimetric analysis, and Raman spectroscopy were employed to validate the quality and structure of the produced graphene materials. The major findings of this study reveal the successful implementation of the methodology, leading to the production of high-quality graphene materials suitable for advanced material applications. Thorough characterization using various advanced techniques validates the structural integrity and purity of the graphene. The economic viability of the process is demonstrated through a comprehensive economic analysis, highlighting the potential for large-scale production. This research contributes to the field of sustainable production of graphene materials by offering a systematic methodology that efficiently transforms spent batteries into valuable graphene resources. Furthermore, the findings not only showcase the potential for upcycling electronic waste but also address the pressing need for environmentally conscious processes in advanced material synthesis.

Keywords: spent primary batteries, spent secondary batteries, graphite extraction, advanced material synthesis, circular economy approach

Procedia PDF Downloads 38

Authors: Nishal Parbhoo, John B. Dewar, Samantha Gildenhuys

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Rotavirus is the major cause of severe gastroenteritis worldwide in children aged 5 and younger. Death rates are high particularly in developing countries. The mature rotavirus is a non-enveloped triple-layered nucleocapsid containing 11 double-stranded RNA segments. Here a global view on the sequence and structure of the three main capsid proteins, VP7, VP6, and VP2 is taken by generating a consensus sequence for each of these rotavirus proteins, for each species obtained from published data of representative rotavirus genotypes from across the world and across species. The degree of conservation between species was represented on homology models for each of the proteins. VP7 shows the highest level of variation with 14 - 45 amino acids showing conservation of less than 60%. These changes are localized to the outer surface which is exposed to antibodies alluding to a possible mechanism in evading the immune system. The middle layer, VP6 shows lower variability with only 14-32 sites having lower than 70% conservation. The inner structural layer made up of VP2 showed the lowest variability with only 1-16 sites having less than 70% conservation across species. The results correlate with proteins’ multiple structural roles. Although the nucleotide sequences vary due to an error-prone replication and lack of proofreading, the corresponding amino acid sequence of VP2, 6 and 7 remains conserved. Sequence conservation maintained for the virus results in stable protein structures, fit for function. This can be exploited in drug design, molecular studies and biotechnological applications.

Keywords: amino acid sequence conservation, capsid protein, protein structure, vaccine candidate

Procedia PDF Downloads 281

Authors: Endrick Barnacin, Jean-Luc Henry, Jimmy Nagau, Jack Molinie

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Palynology is a field of interest in many disciplines due to its multiple applications: chronological dating, climatology, allergy treatment, and honey characterization. Unfortunately, the analysis of a pollen slide is a complicated and time consuming task that requires the intervention of experts in the field, which are becoming increasingly rare due to economic and social conditions. That is why the need for automation of this task is urgent. A lot of studies have investigated the subject using different standard image processing descriptors and sometimes hand-crafted ones.In this work, we make a comparative study between classical feature extraction methods (Shape, GLCM, LBP, and others) and Deep Learning (CNN, Autoencoders, Transfer Learning) to perform a recognition task over 80 regional pollen species. It has been found that the use of Transfer Learning seems to be more precise than the other approaches

Keywords: pollens identification, features extraction, pollens classification, automated palynology

Procedia PDF Downloads 120

Authors: Othman E. Othman, Hassan R. Darwish, Amira M. Nowier

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Lactoferrin (LF) is a multifunctional protein involved in economically production traits like milk protein composition and skeletal structure in small ruminants including sheep and goat. So, LF gene - with its genetic polymorphisms associated with production traits - is considered a candidate genetic marker used in marker-assisted selection in goats. This study aimed to identify the different alleles and genotypes of this gene in three Egyptian goat breeds using PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing. Genomic DNA was extracted from 120 animals belonging to Barki, Zaraibi, and Damascus goat breeds. Using specific primers, PCR amplified 247-bp fragments from exon 2 of LF goat gene. The PCR products were subjected to Single-Strand Conformation Polymorphism (SSCP) technique. The results showed the presence of two genotypes GG and AG in the tested animals. The frequencies of both genotypes varied among the three tested breeds with the highest frequencies of GG genotype in all tested goat breeds. The sequence analysis of PCR products representing these two detected genotypes declared the presence of an SNP (single nucleotide polymorphisms) substitution (G/A) among G and A alleles of this gene. The association between different LF genotypes and milk composition as well as body measurement was estimated. The comparison showed that the animals possess AG genotypes are superior over those with GG genotypes for different parameters of milk protein compositions and skeletal structures. This finding declared that allele A of LF gene is considered the promising marker for the productive traits in goat. In conclusion, the Egyptian goat breeds will be needed to enhance their milk protein composition and growth trait parameters through the increasing of allele A frequency in their herds depending on the superior production traits of this allele in goats.

Keywords: lLactoferrin gene, PCR-SSCP, SNPs, Egyptian goat

Procedia PDF Downloads 139

Authors: Malgorzata Krysiak, Anna Wegrzyn, Maciej Garstka, Radoslaw Mazur

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Under constantly fluctuating environmental conditions, the thylakoid membrane protein network evolved the ability to dynamically respond to changing biotic and abiotic factors. One of the most important protective mechanism is rearrangement of the chlorophyll-protein (CP) complexes, induced by protein phosphorylation. In a temperate climate, low temperature is one of the abiotic stresses that heavily affect plant growth and productivity. The aim of this study was to determine the role of LHCII antenna complex phosphorylation in the dark-chilling response. The study included an experimental model based on dark-chilling at 4 °C of detached chilling sensitive (CS) runner bean (Phaseolus coccineus L.) and chilling tolerant (CT) garden pea (Pisum sativum L.) leaves. This model is well described in the literature as used for the analysis of chilling impact without any additional effects caused by light. We examined changes in thylakoid membrane protein phosphorylation, interactions between phosphorylated LHCII (P-LHCII) and CP complexes, and their impact on the dynamics of photosystem II (PSII) under dark-chilling conditions. Our results showed that the dark-chilling treatment of CS bean leaves induced a substantial increase of phosphorylation of LHCII proteins, as well as changes in CP complexes composition and their interaction with P-LHCII. The PSII photochemical efficiency measurements showed that in bean, PSII is overloaded with light energy, which is not compensated by CP complexes rearrangements. On the contrary, no significant changes in PSII photochemical efficiency, phosphorylation pattern and CP complexes interactions were observed in CT pea. In conclusion, our results indicate that different responses of the LHCII phosphorylation to chilling stress take place in CT and CS plants, and that kinetics of LHCII phosphorylation and interactions of P-LHCII with photosynthetic complexes may be crucial to chilling stress response. Acknowledgments: presented work was financed by the National Science Centre, Poland grant No.: 2016/23/D/NZ3/01276

Keywords: LHCII, phosphorylation, chilling stress, pea, runner bean

Procedia PDF Downloads 120

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

Procedia PDF Downloads 300

Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

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A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

Procedia PDF Downloads 251

Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

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Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.

Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome

Procedia PDF Downloads 187