Search results for: iodine affinity

Authors: Khaled S. Al Salhen

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Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (Km = 78 ± 7.6 µM) for hepatic aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: aldehyde oxidase, fish, phenanthridine, specificity

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Authors: Natasha Mandal, Rakesh Singh Moirangthem

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The present work reports a theoretical model to develop a plasmonic biosensor for the detection of glucose concentrations in human blood and urine as the abnormality of glucose label is the major cause of diabetes which becomes a life-threatening disease worldwide. This study is based on the surface plasmon resonance (SPR) sensor applications which is a well-established, highly sensitive, label-free, rapid optical sensing tool. Here we have introduced a sandwich assay of two dielectric spacer layers of MgF2 and BaTiO3which gives better performance compared to commonly used SiO2 and TiO2 dielectric spacers due to their low dielectric loss and higher refractive index. The sensitivity of our proposed sensor was found as 3242 nm/RIU approximately, with an excellent linear response of 0.958, which is higher than the conventional single-layer Au SPR sensor. Further, the sensitivity enhancement is also optimized by coating a few layers of two-dimensional (2D) nanomaterials (e.g., Graphene, h-BN, MXene, MoS2, WS2, etc.) on the sensor chip. Hence, our proposed SPR sensor has the potential for the detection of glucose concentration in blood and urine with enhanced sensitivity and high affinity and could be utilized as a reliable platform for the optical biosensing application in the field of medical diagnosis.

Keywords: biosensor, surface plasmon resonance, dielectric spacer, 2D nanomaterials

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Authors: Oktira Roka Aji, Maelita R. Moeis, Ihsanawati, Ernawati A. Giri-Rachman

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Globally, tuberculosis (TB) remains a leading cause of death. The emergence of multidrug-resistant strains and extensively drug-resistant strains have become a major public concern. One of the potential candidates for drug target is the cytoplasmic domain of PhoR Histidine Kinase, a part of the Two Component System (TCS) PhoR-PhoP in Mycobacterium tuberculosis (Mtb). TCS PhoR-PhoP relay extracellular signal to control the expression of 114 virulent associated genes in Mtb. The 3D structure of PhoR cytoplasmic domain is needed to screen novel drugs using structure based drug discovery. The PhoR cytoplasmic domain from Mtb H37Rv was overexpressed in E. coli BL21(DE3), then purified using IMAC Ni-NTA Agarose his-tag affinity column and DEAE-ion exchange column chromatography. The molecular weight of the purified protein was estimated to be 37 kDa after SDS-PAGE analysis. This sample was used for pre-crystallization screening by applying sitting drop vapor diffusion method using Natrix (HR2-116) 48 solutions crystal screen kit at 25ºC. Needle-like crystals were observed after the seventh day of incubation in test solution No.47 (0.1 M KCl, 0.01 M MgCl2.6H2O, 0.05 M Tris-Cl pH 8.5, 30% v/v PEG 4000). Further testing is required for confirming the crystal.

Keywords: tuberculosis, two component system, histidine kinase, needle-like crystals

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Authors: Jain Jyoti, Jain Shorab, Sinha Shishir

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In the current scenario, development of new biodegradable composites with the reinforcement of some plant derived natural fibers are in major research concern. Abundant quantity of these natural plant derived fibers including sisal, ramp, jute, wheat straw, pine, pineapple, bagasse, etc. can be used exclusively or in combination with other natural or synthetic fibers to augment their specific properties like chemical, mechanical or thermal properties. Among all natural fibers, wheat straw, bagasse, kenaf, pineapple leaf, banana, coir, ramie, flax, etc. pineapple leaf fibers have very good mechanical properties. Being hydrophilic in nature, pineapple leaf fibers have very less affinity towards all types of polymer matrixes. Not much work has been carried out in this area. Surface treatments like alkaline treatment in different concentrations were conducted to improve its compatibility towards hydrophobic polymer matrix. Pineapple leaf fiber epoxy composites have been prepared using hand layup method. Effect of variation in fiber loading up to 20% in epoxy composites has been studied for mechanical properties like tensile strength and flexural strength. Analysis of fiber morphology has also been studied using FTIR, XRD. SEM micrographs have also been studied for fracture surface.

Keywords: composite, mechanical, natural fiber, pineapple leaf fiber

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Authors: K. Haggag, N. S. Elshemy

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Due to limited fossil resource and an increased need for environmentally friendly, sustainable technologies, the importance of using renewable feed stocks in textile industry area will increase in the decades to come. This research highlights some of the perspectives in this area. Alkyd resins for high characterization and reactive properties, completely based on commercially available renewable resources (sunflower and/or soybean oil) were prepared and characterized. In this work, we present results on the synthesis of various alkyd resins according to the alcoholysis – polyesterification process under different preparation conditions using a microwave synthesis as energy source to determine suitable reaction conditions. Effects of polymerization parameters, such as catalyst ratio, reaction temperature and microwave power level have been studied. The prepared binder was characterized via FT-IR, scanning electron microscope (SEM) and transmission electron microscope (TEM), in addition to acid value (AV), iodine value (IV), water absorbance, weight loss, and glass transition temperature. The prepared binder showed high performance physico-mechanical properties. TEM analysis showed that the polymer latex nanoparticle within range of 20–200 nm. The study involved the application of the prepared alkyd resins as binder for pigment printing process onto cotton fabric by using a flat screen technique and the prints were dried and thermal cured. The optimum curing conditions were determined, color strength and fastness properties of pigment printed areas to light, washing, perspiration and crocking were evaluated. The rheological properties and apparent viscosity of prepared binders were measured in addition roughness of the prints was also determined.

Keywords: nano-binder, microwave heating, renewable resource, alkyd resins, sunflower oil, soybean oil

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Authors: Cíntia Martins Sanches

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In the first part of Phoenissae of Seneca, Antigone is a guide to Oedipus, and they leave Thebes: he is blind searching for death (inflicting the punishment himself wished on the killer of Laius, ie exile and death); she is trying to convince him to give up such punishment and bring him back to Thebes. Concerning Oedipus lines, we observed a high frequency of Latin neuter in the treatment the protagonist gave to his daughter Antigone. We considered in this study that such frequency may be related to the sanctification of the daughter, who is seen by him as an enlightened being and without defects, free of the human condition (which takes on the existence of failures by essence). This study, thus, puts forward an analysis of the passages the said feature is present, relating them to the effect of meaning found in each occurrence. As part of a doctorate, this study investigates the stylistic idiom of Seneca in the Oedipus and Phoenissae tragedies, aiming at translating both tragedies expressively. The concept of stylistic idiom concerns the stylistic affinity required for a translation to be equivalent to the source text. In this wise, this study inquires into how the Latin text is organized poetically, pointing out the expressive features frequently appearing in both dramas. The method we used is based on the Semiotics theory — observing how connotation, ie a language use in which prevails the poetic function, naturally polysemous, acts to achieve each expressive effect.

Keywords: antigone, neuter, Oedipus, Phoenissae, Seneca

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Authors: H. Mokaddem, D. Miroud, N. Azouaou, F. Si-Ahmed, Z. Sadaoui

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The synthesis and characterization of activated carbon from local lignocellulosic precursor (Algerian alfa) was carried out for the removal of cationic dyes from aqueous solutions. The effect of the production variables such as impregnation chemical agents, impregnation ratio, activation temperature and activation time were investigated. Carbon obtained using the optimum conditions (CaCl2/ 1:1/ 500°C/2H) was characterized by various analytical techniques scanning electron microscopy (SEM), infrared spectroscopic analysis (FTIR) and zero-point-of-charge (pHpzc). Adsorption tests of methylene blue on the optimal activated carbon were conducted. The effects of contact time, amount of adsorbent, initial dye concentration and pH were studied. The adsorption equilibrium examined using Langmuir, Freundlich, Temkin and Redlich–Peterson models reveals that the Langmuir model is most appropriate to describe the adsorption process. The kinetics of MB sorption onto activated carbon follows the pseudo-second order rate expression. The examination of the thermodynamic analysis indicates that the adsorption process is spontaneous (ΔG ° < 0) and endothermic (ΔH ° > 0), the positive value of the standard entropy shows the affinity between the activated carbon and the dye. The present study showed that the produced optimal activated carbon prepared from Algerian alfa is an effective low-cost adsorbent and can be employed as alternative to commercial activated carbon for removal of MB dye from aqueous solution.

Keywords: activated carbon, adsorption, cationic dyes, Algerian alfa

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Authors: S. Kerakra, S. Bouhelal, M. Poncot

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The aim of this study is to give a comprehensive overview of the effect of short hollow and solid recycled polyethylene terephthalate (PET) fibers in different breaking tenacities reinforced isotactic polypropylene (iPP) composites on the mechanical and morphological properties. Composites of iPP/3, 7and 10 wt% of solid and hollow recycled PET fibers were prepared by batched melt mixing in a Brabender. The incorporation of solid recycled-PET fibers in isotactic polypropylene increase Young’s modulus of iPP relatively, meanwhile it increased proportionally with hollow fibers content. An improvement of the storage modulus, and a shift up in glass transition temperatures of hollow fibers/iPP composites was determined by DMA results. The morphology of composites was determined by scanning electron microscope (SEM) and optical polarized microscopy (OM) showing a good dispersion of the hollow fibers. Also, their flexible aspect (folding, bending) was observed. But, one weak interaction between the polymer/fibers phases was shown. Polymers can be effectively reinforced with short hollow recycled PET fibers due to their characteristics like recyclability, lightweight and the flexible aspect, which allows the absorbance of the energy of a striker with a minimum damage of the matrix. Aiming to improve the affinity matrix–recycled hollow PET fibers, it is suggested the addition of compatibilizers, as maleic anhydride.

Keywords: isotactic polypropylene, hollow recycled PET fibers, solid recycled-PET fibers, composites, short fiber, scanning electron microscope

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Authors: Sanja O. Podunavac-Kuzmanović, Lidija R. Jevrić, Strahinja Z. Kovačević

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In last decade, metal-organic complexes have captured intensive attention because of their wide range of biological activities such as antibacterial, antifungal, anticancerous, antimicrobial and antiHIV. Therefore, it is of great importance for the development of coordination chemistry to explore the assembly of functional organic ligands with metal ion and to investigate the relationship between the structure and property. In view of our studies, we reasoned that benzimidazoles complexed to metal ions could act as a potent antibacterial agents. Thus, we have bioassayed the inhibitory potency of benzimidazoles and their metal salts (Co or Ni) against Gram negative bacteria Escherichia coli. In order to validate our in vitro study, we performed in silico studies using molecular docking software’s. The investigated compounds and their metal complexes (Co, Ni) showed good antibacterial activity against Escherichia coli. In silico docking studies of the synthesized compounds suggested that complexed benzimidazoles have a greater binding affinity and enhanced antibacterial activity in comparison with noncomplexed ligands. In view of their enhanced inhibitory properties we propose that the studied complexes can be used as potential pharmaceuticals. This study is financially supported by COST action CM1306 and the project No. 114-451-347/2015-02, financially supported by the Provincial Secretariat for Science and Technological Development of Vojvodina.

Keywords: benzimidazoles, complexes, antibacterial, Escherichia coli, metal

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Authors: Dalia. G. Aseel

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Begomoviruses are economically important plant viruses that infect dicotyledonous plants and exclusively transmitted by the whitefly Bemisia tabaci. Here, replicative form was isolated from Okra, Cotton, Tomato plants and whitefly infected with Begomoviruses. Using coat protein specific primers (AV1), the viral infection was verified with amplicon at 450 bp. The sequence of OLCuV-AV1 gene was recorded and received an accession number (FJ441605) from Genebank. The phylogenetic tree of OLCuV was closely related to Okra leaf curl virus previously isolated from Cameroon and USA with nucleotide sequence identity of 92%. The protein purification was carried out using His-Tag methodology by using Affinity Chromatography. The purified protein was separated on SDS-PAGE analysis and an enriched expected size of band at 30 kDa was observed. Furthermore, RAPD and SDS-PAGE were used to detect genetic variability between different hosts of okra leaf curl virus (OLCuV), cotton leaf curl virus (CLCuV), tomato yellow leaf curl virus (TYLCuV) and the whitefly vector. Finally, the present study would help to understand the relationship between the whitefly and different economical crops in Egypt.

Keywords: okra leaf curl virus, AV1 gene, sequencing, phylogenetic, cloning, purified protein, genetic diversity and viral proteins

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Authors: Angelis P. Barlampas

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Purpose or Learning Objective It is well known that previous examinations play a major role in futural diagnosis, thus avoiding unnecessary new exams that cost in time and money both for the patient and the health system. A case is presented in which past patient’s results, in combination with the least needed new tests, give an easy final diagnosis. Methods or Background A middle aged man visited the emergency department complaining of hard controlled, persisting fever for the last few days. Laboratory tests showed an elevated number of white blood cells with neutrophil shift and abnormal CRP. The patient was admitted to hospital a month ago for continuing lungs symptomatology after a recent covid-19 infection. Results or Findings Computed tomography scanning showed a solid mass with spiculating margins in right lower lobe. After intravenous iodine contrast administration, there was mildly peripheral enhancement and eccentric non enhancing area. A pneumonic cancer was suspected. Comparison with the patient’s latest computed tomography revealed no mass in the area of interest but only signs of recent post covid-19 lung parenchyma abnormalities. Any new mass that appears in a month’s time span can not be a cancer but a benign lesion. It was obvious that an abscess was the most suitable explanation. The patient was admitted to hospital, and antibiotic therapy was given, with very good results. After a few days, the patient was afebrile and in good condition. Conclusion In this case , a PET scan or a biopsy was avoided, thanks to the patient’s medical history and the availability of previous examinations. It is worthy encouraging the patients to keep their medical records and organizing more efficiently the health system with the current technology of archiving the medical examinations, too.

Keywords: covid-19, chest ct, cancer, abscess, fever

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Authors: Abujnah Dukali

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Natural honey was assessed in cell culture system for its anticancer activity. Human leukemic cell line HL 60 was treated with honey and cultured for 5 days and cytotoxicity was calculated by MTT assay. Honey showed cytotoxicity with CC50 value of 174.20 µg/ml. Radical modulation activities was assessed by lipid peroxidation assay using egg lecithin. Honey showed antioxidant activity with EC50 value of 159.73 µg/ml. In addition, treatment with HL60 cells also resulted in nuclear DNA fragmentation, as seen in agarose gel electrophoresis. This is a hallmark of cells undergoing apoptosis. Confirmation of apoptosis was performed by staining the cells with Annexin V and FACS analysis. Apoptosis is an active, genetically regulated disassembly of the cell form within. Disassembly creates changes in the phospholipid content of the cytoplasmic membrane outer leaflet. Phosphatidylserine (PS) is translocated from the inner to the outer surface of the cell for phagocytic cell recognition. The human anticoagulant, annexin V, is a Ca2+-dependent phospholipid protein with a high affinity for PS. Annexin V labeled with fluorescein can identify apoptotic cells in the population It is a confirmatory test for apoptosis. Annexin V-positive cells were defined as apoptotic cells. Since honey shows both antioxidant activity and cytotoxicity at almost the same concentration, it can prevent the free radical induced cancer as prophylactic agent and kill the cancer cells by apoptotic process as a chemotherapeutic agent. Everyday intake of honey can prevent the cancer induction.

Keywords: anticancer, cells, DNA, honey

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Authors: Henri El Zakhem, Anthony Dahdah, Lara Frangieh, Jessica Koura

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Nowadays, the removal of whey produced in the dairy processes has been the most important problem in the dairy industry. Every year, about 47% of the 115 million tons of whey produced world-wide are disposed in the environment. Whey is a nutritious liquid, containing whey proteins (β-lactoglobulin, α-lactalbumin, immunoglobulin-G, proteose pepton), lactose, vitamins (B5, B2, C, and B6), minerals (Calcium, Magnesium, Phosphorous, Potassium, Chloride, and Sodium), and trace elements (Zinc, Iron, Iodine, and Copper). The first objective was to increase the economical and commercial value of whey which is considered as by-product. The second objective of this study was to formulate a new baby food with good nutritional, sensory and storage properties and acceptable to consumers using the cheese whey. The creation of the new product must pass through the following stages: idea stage, development stage which includes the business planning and the product development prototype, packaging stage, production stage, test marketing stage, quality control/sanitation. Three types of whey-based food were selected and prepared by mixing whey and apple, whey and banana as well as whey, apple, and banana.To compile with the recommended dietary allowances (RDA) and adequate intakes (AI) for vitamins and minerals, each sample is formed from 114g of sliced and smashed fruits mixed with 8 mL of whey. Mixtures are heated to 72oC for 15 seconds, and filled in pasteurized jars. Jars were conserved at 4oC. Following the experimental part, sensory evaluation made by an experienced panel took place. Hedonic tests results show that the mixture of whey, apple, and banana has the most delicious and sweetness taste followed by the mixture of whey and banana, and finally the mixture of whey and apple. This study was concluded with a managerial and engineering study that reveals that the project is economically profitable to be executed in Lebanon.

Keywords: baby food, by-product, cheese whey, formulation

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Authors: Dovilė Bačėninaitė, Karina Džermeikaitė, Justinas Kirvela, Ramūnas Antanaitis

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Bacteria are often present in the mouth of cattle. Some of them can cause abscesses. Starting with severe swelling of the mouth, muscle spasm, or locked jaw, it can lead to inability to open its mouth, move the neck, cause pain while eating. While the calf is unable to eat properly, it becomes more susceptible to infectious diseases, lower weight gain can be observed. Abscesses can be considered as a continuum of oral disease, whereby early stages of the lumpy jaw could proceed from gingivitis to periodontal disease. In the event of tissue damage, bacteria can enter the bloodstream, even cause sepsis. The most common lesions occur when animals eat sharp grass, coarse fodder, sharp, piercing foreign bodies (this is especially common for calves when they are trying to eat inedible objects). A crossbred Holstein calf presented with a history of proliferative outgrowth in the mandibular region. On clinical examination, needle aspiration, mandibular swelling revealed sticky, white curd-like fluid containing. Pus bacteriology revealed gram-negative cocci. They were sensitive to amoxicillin, cephalexin, enrofloxacin, ceftiofur. Blood morphology was in physiological ranges. The calf was treated surgically. The growth was excised, the puss drained and the wound was flushed with potassium permanganate solution (0,01%). A week after clinical surgery examination was performed. The swelling was decreased. Superficial bacterial infections are often associated with poor hygiene, which should be improved before treatment is commenced. Clipping away dirty hair and gently washing affected areas of skin daily with solutions such as povidone-iodine, potassium permanganate is effective. Appropriate antibiotic therapy, based on sensitivity testing, may be used where there is evidence of systemic illness.

Keywords: calf, abscess, lumpy jaw, pus, Streptococcus, Staphylococcus, Actinobacillus, infection

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Authors: Sakina Fatima, Joseph-Patrick W. E. Clarke, Patricia A. Thibault, Subha Kalyaanamoorthy, Michael Levin, Aravindhan Ganesan

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Heteronuclear ribonucleoprotein (hnRNPA1 or A1) is associated with the pathology of different diseases, including neurological disorders and cancers. In particular, the aggregation and dysfunction of A1 have been identified as a critical driver for neurodegeneration (NDG) in Multiple Sclerosis (MS). Structurally, A1 includes a low-complexity domain (LCD) and two RNA-recognition motifs (RRMs), and their interdomain coordination may play a crucial role in A1 aggregation. Previous studies propose that RNA-inhibitors or nucleoside analogs that bind to RRMs can potentially prevent A1 self-association. Therefore, molecular-level understanding of the structures, dynamics, and nucleotide interactions with A1 RRMs can be useful for developing therapeutics for NDG in MS. In this work, a combination of computational modelling and biochemical experiments were employed to analyze a set of RNA-A1 RRM complexes. Initially, the atomistic models of RNA-RRM complexes were constructed by modifying known crystal structures (e.g., PDBs: 4YOE and 5MPG), and through molecular docking calculations. The complexes were optimized using molecular dynamics simulations (200-400 ns), and their binding free energies were computed. The binding affinities of the selected complexes were validated using a thermal shift assay. Further, the most important molecular interactions that contributed to the overall stability of the RNA-A1 RRM complexes were deduced. The results highlight that adenine and guanine are the most suitable nucleotides for high-affinity binding with A1. These insights will be useful in the rational design of nucleotide-analogs for targeting A1 RRMs.

Keywords: hnRNPA1, molecular docking, molecular dynamics, RNA-binding proteins

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Authors: Sharifah Mousli, Sona Taheri, Jiayuan He

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Autism spectrum disorder (ASD) is a complex developmental condition involving persistent difficulties with social communication, restricted interests, and repetitive behavior. The challenges associated with ASD can interfere with an affected individual’s ability to function in social, academic, and employment settings. Although there is no effective medication known to treat ASD, to our best knowledge, early intervention can significantly improve an affected individual’s overall development. Hence, an accurate diagnosis of ASD at an early phase is essential. The use of machine learning approaches improves and speeds up the diagnosis of ASD. In this paper, we focus on the application of unsupervised clustering methods in ASD as a large volume of ASD data generated through hospitals, therapy centers, and mobile applications has no pre-existing labels. We conduct a comparative analysis using seven clustering approaches such as K-means, agglomerative hierarchical, model-based, fuzzy-C-means, affinity propagation, self organizing maps, linear vector quantisation – as well as the recently developed optimization-based clustering (COMSEP-Clust) approach. We evaluate the performances of the clustering methods extensively on real-world ASD datasets encompassing different age groups: toddlers, children, adolescents, and adults. Our experimental results suggest that the COMSEP-Clust approach outperforms the other seven methods in recognizing ASD with well-separated clusters.

Keywords: autism spectrum disorder, clustering, optimization, unsupervised machine learning

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Authors: Shashank Kumar, Mansi Chandra, Ujjawal Singh, Parth Gupta, Rishi Ram, Arnab Sarkar

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Milk is one of the basic and primary sources of food and energy as we start consuming milk from birth. Hence, milk quality and purity and checking the concentration of its constituents become necessary steps. Considering the importance of the purity of milk for human health, the following study has been carried out to simultaneously detect and quantify the different adulterants like urea and starch in milk with the help of a paper-based microfluidic device integrated with a smartphone. The detection of the concentration of urea and starch is based on the principle of colorimetry. In contrast, the fluid flow in the device is based on the capillary action of porous media. The microfluidic channel proposed in the study is equipped with a specialized detection zone, and it employs a colorimetric indicator undergoing a visible color change when the milk gets in touch or reacts with a set of reagents which confirms the presence of different adulterants in the milk. In our proposed work, we have used iodine to detect the percentage of starch in the milk, whereas, in the case of urea, we have used the p-DMAB. A direct correlation has been found between the color change intensity and the concentration of adulterants. A calibration curve was constructed to find color intensity and subsequent starch and urea concentration. The device has low-cost production and easy disposability, which make it highly suitable for widespread adoption, especially in resource-constrained settings. Moreover, a smartphone application has been developed to detect, capture, and analyze the change in color intensity due to the presence of adulterants in the milk. The low-cost nature of the smartphone-integrated paper-based sensor, coupled with its integration with smartphones, makes it an attractive solution for widespread use. They are affordable, simple to use, and do not require specialized training, making them ideal tools for regulatory bodies and concerned consumers.

Keywords: paper based microfluidic device, milk adulteration, urea detection, starch detection, smartphone application

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Authors: Waranya Imprasittichai, Patsarawadee Paojinda, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. In this study, we constructed gene fusions of Plasmodium falciparum OMPDC-OPRT (1,836 bp) in pTrcHisA vector and expressed as an 6xHis-tag bifunctional protein in three Escherichia coli strains (BL21, Rosetta, TOP10) at 18 °C, 25 °C and 37 °C. The recombinant bifunctional protein was partially purified by Ni-Nitrilotriacetic acid-affinity chromatography. Specific activities of OPRT and OMPDC domains in the bifunctional enzyme expressed in E. coli TOP10 cells were approximately 3-4-fold higher than those in BL21 cells. There were no enzymatic activities when the construct vector expressed in Rosetta cells. Maximal expression of the fused gene was observed at 18 °C and the bifunctional enzyme had specific activities of OPRT and OMPDC domains in a ratio of 1:2. These results provide greater yields and better catalytic activities of the bifunctional OMPDC-OPRT enzyme for further purification and kinetic study.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: O’Day Luke

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Purpose: To validate virtual non-contrast reconstructions generated from dual-layer spectral computed tomography (DL-CT) data as an alternative for the acquisition of a dedicated true non-contrast dataset during multiphase contrast studies. Material and methods: Thirty-three patients underwent a routine multiphase clinical CT examination, using Dual-Layer Spectral CT, from March to August 2021. True non-contrast (TNC) and virtual non-contrast (VNC) datasets, generated from both portal venous and arterial phase imaging were evaluated. For every patient in both true and virtual non-contrast datasets, a region-of-interest (ROI) was defined in aorta, liver, fluid (i.e. gallbladder, urinary bladder), kidney, muscle, fat and spongious bone, resulting in 693 ROIs. Differences in attenuation for VNC and TNV images were compared, both separately and combined. Consistency between VNC reconstructions obtained from the arterial and portal venous phase was evaluated. Results: Comparison of CT density (HU) on the VNC and TNC images showed a high correlation. The mean difference between TNC and VNC images (excluding bone results) was 5.5 ± 9.1 HU and > 90% of all comparisons showed a difference of less than 15 HU. For all tissues but spongious bone, the mean absolute difference between TNC and VNC images was below 10 HU. VNC images derived from the arterial and the portal venous phase showed a good correlation in most tissue types. The aortic attenuation was somewhat dependent however on which dataset was used for reconstruction. Bone evaluation with VNC datasets continues to be a problem, as spectral CT algorithms are currently poor in differentiating bone and iodine. Conclusion: Given the increasing availability of DL-CT and proven accuracy of virtual non-contrast processing, VNC is a promising tool for generating additional data during routine contrast-enhanced studies. This study shows the utility of virtual non-contrast scans as an alternative for true non-contrast studies during multiphase CT, with potential for dose reduction, without loss of diagnostic information.

Keywords: dual-layer spectral computed tomography, virtual non-contrast, true non-contrast, clinical comparison

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Authors: Ergun Sakalar, Kubra Bilgic

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In the last decades, real-time PCR has become a reliable tool preferred to use in many laboratories for pathogen detection. This technique allows for monitoring target amplification via fluorescent molecules besides admit of quantitative analysis by enabling of convert outcomes of thermal cycling to digital data. Sensitivity and traceability of real-time PCR are based on measuring of fluorescence that appears only when fluorescent reporter dye bound to specific target DNA.The fluorescent reporter systems developed for this purpose are divided into two groups. The first group consists of intercalator fluorescence dyes such as SYBR Green, EvaGreen which binds to double-stranded DNA. On the other hand, the second group includes fluorophore-labeled oligonucleotide probes that are separated into three subgroups due to differences in mechanism of action; initial primer-probes such as Cyclicons, Angler®, Amplifluor®, LUX™, Scorpions, and the second one hydrolysis probes like TaqMan, Snake assay, finally hybridization probes, for instance, Molecular Beacons, Hybprobe/FRET, HyBeacon™, MGB-Eclipse, ResonSense®, Yin-Yang, MGB-Pleiades. In addition nucleic acid analogues, an increase of probe affinity to target site is also employed with fluorescence-labeled probes. Consequently, abundant real-time PCR detection chemistries are chosen by researcher according to the field of application, mechanism of action, advantages, and proper structures of primer/probes.

Keywords: fluorescent dye, food safety, molecular probes, nucleic acid analogues

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Authors: Zhuoyuan Wang, Benson T. H. Lim, Imriyas Kamardeen

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Effective service quality management has long been touted as a means of improving project and organisational performance. Particularly, in construction projects, main contractors are often seen as a broker between clients and subcontractors, and their service quality is thus associated with the overall project affinity and outcomes. While a considerable amount of research has focused on the aspect of clients-main contractors, very little research has been done to explore the effect of contractors’ service quality on subcontractors’ behaviours and so project outcomes. In addressing this gap, this study surveyed 97 subcontractors in the Chinese Construction industry and data was analysed using the Partial Least Square (PLS) Structural Equation Modelling (SEM) technique. The overall findings reveal that subcontractors categorised main contractors’ service quality into three dimensions: assurance; responsiveness; reliability and empathy. Of these, it is found that main contractors’ ‘assurance’ and ‘responsiveness’ positively influence subcontractors’ intention to engage in contractual behaviours. The results further show that the subcontractors’ intention to engage in organizational citizenship behaviours is associated with how flexible and committed the main contractors are in reliability and empathy. Collectively, both subcontractors’ contractual and organizational citizenship behaviours positively influence the overall project outcomes. In conclusion, the findings inform contractors different strategies towards managing and gaining subcontractors’ behaviour commitment in a socially connected, yet complex and uncertain, business environment.

Keywords: construction firms, organisational citizenship behaviour, service quality, social exchange theory

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Authors: Hyunuk Kim, Yang No Yun, Muhammad Sohail, Jong-Ho Moon, Young Cheol Park

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Metal-organic frameworks (MOFs), which are emerging absorbents assembled from metal ions and organic ligands, have attracted attention for their permanent porosity and design of tunable pore size. These microporous materials showed interesting properties for CO₂ storage and separation. In particular, MOFs with high surface area and open metal sites showed the remarkable adsorption capacity and selectivity for CO₂. [Mg₂ (DOBDC)] (DOBDC = 2,5-dioxidobenzene-1,4-dicarboxylate) (MOF-74 or CPO-27) is a well-known absorbent showing an exceptionally high CO₂ sorption capacity at low partial pressure and room temperature. In this work, we synthesized [M₂(DOBDC)(DMF)₂] (M = Mg, Co, Ni) and determined their single-crystal structures by X-ray crystallography. The removal of coordinated guest molecules generates Lewis acidic sites and showed high CO₂ adsorption affinity. Both CO₂ adsorption capacity and surface area are much higher than reported values in literature. To fabricate MMMs, microcrystalline [M₂ (DOBDC)(DMF)₂] was synthesized by microwave reaction and dispersed in PDMS solution. The MMMs with a various amount of [M₂ (DOBDC)(DMF) ₂] in PDMS were fabricated by a solution casting method. [M₂ (DOBDC)(DMF)₂]@PDMS membrane showed higher CO2 permeability and CO₂/N₂ selectivity than those of PDMS. Therefore, we believe that MMMs combining polymer and MOFs provide new materials for CO₂ separation technology.

Keywords: metal-organic frameworks, mixed matrix membrane, CO2/N2 separation, polydimethylsiloxane (PDMS)

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Authors: Juan Jose Filgueira, Daniela Londono-Serna, Liliana Maria Hoyos

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Carnation (Dianthus caryophyllus) is one of the most important products of exportation in the floriculture industry worldwide. Fusariosis is the disease that causes the highest losses on farms, in particular the one produced by Fusarium oxysporum f.sp. dianthi, called vascular wilt. Gene identification and metabolic routes of the genes that participate in the building of the plant response to Fusarium are some of the current targets in the carnation breeding industry. The techniques for the identifying of resistant genes in the plants, is the analysis of the transcriptome obtained during the host-pathogen interaction. In this work, we report the cell transcriptome of different varieties of carnation that present differential response from Fusarium oxysporum f.sp. dianthi attack. The cells of the different hybrids produced in the outbreeding program were cultured in vitro and elicited with the parasite in a dual culture. The isolation and purification of mRNA was achieved by using affinity chromatography Oligo dT columns and the transcriptomes were obtained by using Illumina NGS techniques. A total of 85,669 unigenes were detected in all the transcriptomes analyzed and 31,000 annotations were found in databases, which correspond to 36.2%. The library construction of genic expression techniques used, allowed to recognize the variation in the expression of genes such as Germin-like protein, Glycosyl hydrolase family and Cinnamate 4-hydroxylase. These have been reported in this study for the first time as part of the response mechanism to the presence of Fusarium oxysporum.

Keywords: Carnation, Fusarium, vascular wilt, transcriptome

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Authors: Strahinja Kovačević, Sanja Podunavac-Kuzmanović, Lidija Jevrić, Milica Karadžić

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The present study is based on the quantitative structure-activity relationship (QSAR) analysis of eighteen compounds with anti-prion activity. The structures and anti-prion activities (expressed in response units, RU%) of the analyzed compounds are taken from CHEMBL database. In the first step of analysis 85 molecular descriptors were calculated and based on them the hierarchical cluster analysis (HCA) and principal component analysis (PCA) were carried out in order to detect potential significant similarities or dissimilarities among the studied compounds. The calculated molecular descriptors were physicochemical, lipophilicity and ADMET (absorption, distribution, metabolism, excretion and toxicity) descriptors. The first stage of the QSAR analysis was simple linear regression modeling. It resulted in one acceptable model that correlates Henry's law constant with RU% units. The obtained 2D-QSAR model was validated by cross-validation as an internal validation method. The validation procedure confirmed the model’s quality and therefore it can be used for prediction of anti-prion activity. The next stage of the analysis of anti-prion activity will include 3D-QSAR and molecular docking approaches in order to select the most promising compounds in treatment of prion diseases. These results are the part of the project No. 114-451-268/2016-02 financially supported by the Provincial Secretariat for Science and Technological Development of AP Vojvodina.

Keywords: anti-prion activity, chemometrics, molecular modeling, QSAR

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Authors: Rohit Singh Dangi, Ravi Kant Pal, Monica Sundd

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Acyl-CoA binding protein (ACBP) is a housekeeping protein which functions as an intracellular carrier of acyl-CoA esters. Given the fact that the amastigote stage (blood stage) of Leishmania depends largely on fatty acids as the energy source, of which a large part is derived from its host, these proteins might have an important role in its survival. In Leishmania major, genome sequencing suggests the presence of six ACBPs, whose function remains largely unknown. For functional and structural characterization, one of the ACBP genes was cloned, and the protein was expressed and purified heterologously. Acyl-CoA ester binding and stoichiometry were analyzed by isothermal titration calorimetry and Dynamic light scattering. Our results shed light on high affinity of ACBP towards longer acyl-CoA esters, such as myristoyl-CoA to arachidonoyl-CoA with single binding site. To understand the binding mechanism & dynamics, Nuclear magnetic resonance assignments of this protein are being done. The protein's crystal structure was determined at 1.5Å resolution and revealed a classical topology for ACBP, containing four alpha-helical bundles. In the binding pocket, the loop between the first and the second helix (16 – 26AA) is four residues longer from other extensively studied ACBPs (PfACBP) and it curls upwards towards the pantothenate moiety of CoA to provide a large tunnel space for long acyl chain insertion.

Keywords: acyl-coa binding protein (ACBP), acyl-coa esters, crystal structure, isothermal titration, calorimetry, Leishmania

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Authors: Milica Karadzic, Lidija Jevric, Sanja Podunavac-Kuzmanovic, Strahinja Kovacevic, Sinisa Markov, Aleksandar Okljesa, Andrea Nikolic, Marija Sakac, Katarina Penov Gasi

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This study considered the microbiological activity determination and molecular docking study for selected steroid derivatives of biomedical importance. Minimal inhibitory concentration (MIC) was determined for steroid derivatives against Staphylococcus aureus using macrodilution method. Some of the investigated steroid derivatives express bacteriostatic effect against Staphylococcus aureus. Molecular docking approaches are the most widely used techniques for predicting the binding mode of a ligand. Molecular docking study was done for steroid derivatives for androgen receptor negative prostate cancer cell line (PC-3) toward Human Cytochrome P450 CYP17A1. The molecules that had the smallest experimental IC50 values confirmed their ability to dock into active place using suitable molecular docking procedure. The binding disposition of those molecules was thoroughly investigated. Microbiological analysis and molecular docking study were conducted with aim to additionally characterize selected steroid derivatives for future investigation regarding their biological activity and to estimate the binding-affinities of investigated derivatives. This article is based upon work from COST Action (TD1305), supported by COST (European Cooperation and Science and Technology).

Keywords: binding affinity, minimal inhibitory concentration, molecular docking, pc-3 cell line, staphylococcus aureus, steroids

Procedia PDF Downloads 340

Authors: Soujanya Pasumarthi

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Inverse docking is a relatively new technique that has been used to identify potential receptor targets of small molecules. Our docking software package MDock is well suited for such an application as it is both computationally efficient, yet simultaneously shows adequate results in binding affinity predictions and enrichment tests. As a validation study, we present the first stage results of an inverse-docking study which seeks to identify potential direct targets of PRIMA-1. PRIMA-1 is well known for its ability to restore mutant p53's tumor suppressor function, leading to apoptosis in several types of cancer cells. For this reason, we believe that potential direct targets of PRIMA-1 identified in silico should be experimentally screened for their ability to inhibitcancer cell growth. The highest-ranked human protein of our PRIMA-1 docking results is oxidosqualene cyclase (OSC), which is part of the cholesterol synthetic pathway. The results of two followup experiments which treat OSC as a possible anti-cancer target are promising. We show that both PRIMA-1 and Ro 48-8071, a known potent OSC inhibitor, significantly reduce theviability of BT-474 breast cancer cells relative to normal mammary cells. In addition, like PRIMA-1, we find that Ro 48-8071 results in increased binding of mutant p53 to DNA in BT- 474cells (which highly express p53). For the first time, Ro 48-8071 is shown as a potent agent in killing human breast cancer cells. The potential of OSC as a new target for developing anticancer therapies is worth further investigation.

Keywords: inverse docking, in silico screening, protein-ligand interactions, molecular docking

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Authors: Claudia Vergara, Oscar Valdes, Jaime Tapia, Leonardo Santos

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The poly(amidoamine) dendrimers (PAMAM) are a class of material introduced by D. Tomalia. Modifications of the PAMAM dendrimer with several functional groups have attracted the attention for new interesting properties and new applications in many fields such as chemistry, physics, biology, and medicine. However, in the last few years, the use of dendrimers in environmental applications has increased due to pollution concerns. In this contribution, we report the synthesis of three new PAMAM derivates modified with asparagine aminoacid supported in cellulose: PG0-Asn (PAMAM-asparagine), PG0-Asn-Trt (with trityl group) and PG0-Asn-Boc-Trt (with tert-butyl oxycarbonyl group). The functionalization of generation 0 PAMAM dendrimer was carried out by amidation reaction by using an EDC/HOBt protocol. In a second step, functionalized dendrimer was covalently supported to the cellulose surface and used to study the capture of uranyl ions from aqueous solution by fluorescence spectroscopy. The structure and purity of the desired products were confirmed by conventional techniques such as FT-IR, MALDI, elemental analysis, and ESI-MS. Batch experiments were carried out to determine the affinity of uranyl ions with the dendrimer in aqueous solution. Firstly, the optimal conditions for uranyl capture were obtained, where the optimum pH for the removal was 6, the contact time was 4 hours, the initial concentration of uranyl was 100 ppm, and the amount of the adsorbent to be used was 2.5 mg. PAMAM significantly increased the capture of uranyl ions with respect to cellulose as the starting substrate, reaching 94.8% of capture (PG0), followed by 91.2% corresponding to PG0-Asn-Trt, then 70.3% PG0-Asn and 24.2% PG0-Asn-Boc-Trt. These results show that the PAMAM dendrimer is a good option to remove uranyl ions from aqueous solutions.

Keywords: asparagine, cellulose, PAMAM dendrimer, uranyl ions

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Authors: M. A. Ferhat, M. N. Boukhatem, F. Chemat

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Eucalyptus essential oil is extracted from Eucalyptus globulus of the Myrtaceae family and is also known as Tasmanian blue gum or blue gum. Despite the reputation earned by aromatic and medicinal plants of Algeria. The objectives of this study were: (i) the extraction of the essential oil from the leaves of Eucalyptus globulus Labill., Myrtaceae grown in Algeria, and the quantification of the yield thereof, (ii) the identification and quantification of the compounds in the essential oil obtained, and (iii) the determination of physical and chemical properties of EGEO. The chemical constituents of Eucalyptus globulus essential oil (EGEO) of Blida origin has not previously been investigated. Thus, the present study has been conducted for the determination of chemical constituents and different physico-chemical properties of the EGEO. Chemical composition of the EGEO, grown in Algeria, was analysed by Gas Chromatography-Mass Spectrometry. The chemical components were identified on the basis of Retention Time and comparing with mass spectral database of standard compounds. Relative amounts of detected compounds were calculated on the basis of GC peak areas. Fresh leaves of E. globulus on steam distillation yielded 0.96% (v/w) of essential oil whereas the analysis resulted in the identification of a total of 11 constituents, 1.8 cineole (85.8%), α-pinene (7.2%), and β-myrcene (1.5%) being the main components. Other notable compounds identified in the oil were β-pinene, limonene, α-phellandrene, γ-terpinene, linalool, pinocarveol, terpinen-4-ol, and α-terpineol. The physical properties such as specific gravity, refractive index and optical rotation and the chemical properties such as saponification value, acid number and iodine number of the EGEO were examined. The oil extracted has been analyzed to have 1.4602-1.4623 refractive index value, 0.918-0.919 specific gravity (sp.gr.), +9 - +10 optical rotation that satisfy the standards stipulated by European Pharmacopeia. All the physical and chemical parameters were in the range indicated by the ISO standards. Our findings will help to access the quality of the Eucalyptus oil which is important in the production of high value essential oils that will help to improve the economic condition of the community as well as the nation.

Keywords: chemical composition, essential oil, eucalyptol, gas chromatography

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Authors: D. Suman, M. Venkateshwara Rao

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Cancer is a disease caused by an uncontrolled division of abnormal cells in a part of the body, which is affecting millions of people leading to death. Even though there have been tremendous developments taken place over the last few decades the effective therapy for cancer is still not a reality. The existing techniques of cancer therapy are chemotherapy and radio therapy which are having their limitations in terms of the side effects, patient discomfort, radiation hazards and the localization of treatment. This paper describes a novel method for cancer therapy by using RF-hyperthermia application of nanoparticles. We have synthesized ferromagnetic nanoparticles and characterized by using XRD and TEM. These nanoparticles after the biocompatibility studies will be injected in to the body with a suitable tracer element having affinity to the specific tumor site. When RF energy is applied to the nanoparticles at the tumor site it produces heat of excess room temperature and nearly 41-45°C is sufficient to kill the tumor cells. We have designed a RF source generator provided with a temperature feedback controller to control the radiation induced temperature of the tumor site. The temperature control is achieved through a negative feedback mechanism of the thermocouple and a relay connected to the power source of the RF generator. This method has advantages in terms of its effect like localized therapy, less radiation, and no side effects. It has several challenges in designing the RF source provided with coils suitable for the tumour site, biocompatibility of the nanomaterials, cooling system design for the RF coil. If we can overcome these challenges this method will be a huge benefit for the society.

Keywords: hyperthermia, cancer therapy, RF source generator, nanoparticles

Procedia PDF Downloads 441