Search results for: recombinant protein processing

Authors: Reham Khalaf-Nazzal, Imad Dweikat, Paula Gimenez, Iker Oyenarte, Alfonso Martinez-Cruz, Domonik Muller

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Metal cation transport mediator (CNNM) gene family comprises 4 isoforms that are expressed in various human tissues. Structurally, CNNMs are complex proteins that contain an extracellular N-terminal domain preceding a DUF21 transmembrane domain, a ‘Bateman module’ and a C-terminal cNMP-binding domain. Mutations in CNNM2 cause familial dominant hypomagnesaemia. Growing evidence highlights the role of CNNM2 in neurodevelopment. Mutations in CNNM2 have been implicated in epilepsy, intellectual disability, schizophrenia, and others. In the present study, we aim to elucidate the function of CNNM2 in the developing brain. Thus, we present the genetic origin of symptoms in two family cohorts. In the first family, three siblings of a consanguineous Palestinian family in which parents are first cousins, and consanguinity ran over several generations, presented a varying degree of intellectual disability, cone-rod dystrophy, and autism spectrum disorder. Exome sequencing and segregation analysis revealed the presence of homozygous pathogenic mutation in the CNNM2 gene, the parents were heterozygous for that gene mutation. Magnesium blood levels were normal in the three children and their parents in several measurements. They had no symptoms of hypomagnesemia. The CNNM2 mutation in this family was found to locate in the CBS1 domain of the CNNM2 protein. The crystal structure of the mutated CNNM2 protein was not significantly different from the wild-type protein, and the binding of AMP or MgATP was not dramatically affected. This suggests that the CBS1 domain could be involved in pure neurodevelopmental functions independent of its magnesium-handling role, and this mutation could have affected a protein partner binding or other functions in this protein. In the second family, another autosomal dominant CNNM2 mutation was found to run in a large family with multiple individuals over three generations. All affected family members had hypomagnesemia and hypermagnesuria. Oral supplementation of magnesium did not increase the levels of magnesium in serum significantly. Some affected members of this family have defects in fine motor skills such as dyslexia and dyslalia. The detected mutation is located in the N-terminal part, which contains a signal peptide thought to be involved in the sorting and routing of the protein. In this project, we describe heterogenous clinical phenotypes related to CNNM2 mutations and protein functions. In the first family, and up to the authors’ knowledge, we report for the first time the involvement of CNNM2 in retinal photoreceptor development and function. In addition, we report the presence of a neurophenotype independent of magnesium status related to the CNNM2 protein mutation. Taking into account the different modes of inheritance and the different positions of the mutations within CNNM2 and its different structural and functional domains, it is likely that CNNM2 might be involved in a wide spectrum of neuropsychiatric comorbidities with considerable varying phenotypes.

Keywords: magnesium transport, autosomal recessive, autism, neurodevelopment, CBS domain

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Authors: Efren Lopez Jimenez, Maria Isabel Cajero, J. Irving-Vasqueza

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The use and processing of digital images have opened up new opportunities for the resolution of problems of various kinds, such as the calculation of different vegetation indexes, among other things, differentiating healthy vegetation from humid vegetation. However, obtaining images from which these indexes are calculated is still the exclusive subject of active research. In the present work, we propose to obtain these images using a low cost embedded system (Raspberry Pi) and its processing, using a set of libraries of open code called OpenCV, in order to obtain the Normalized Red-Green Difference Index (NGRDI).

Keywords: Raspberry Pi, vegetation index, Normalized Red-Green Difference Index (NGRDI), OpenCV

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Authors: Sarah Gaßmeyer, Nadine Hülsemann, Raphael Stoll, Kenji Miyamoto, Robert Kourist

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Enzymatic racemization allows the smooth interconversion of stereocenters under very mild reaction conditions. Racemases find frequent applications in deracemization and dynamic kinetic resolutions. Arylmalonate decarboxylase (AMDase) from Bordetella Bronchiseptica has high structural similarity to amino acid racemases. These cofactor-free racemases are able to break chemically strong CH-bonds under mild conditions. The racemase-like catalytic machinery of mutant G74C conveys it a unique activity in the racemisation of pharmacologically relevant derivates of 2-phenylpropionic acid (profenes), which makes AMDase G74C an interesting object for the mechanistic investigation of cofactor-independent racemases. Structure-guided protein engineering achieved a variant of this unique racemase with 40-fold increased activity in the racemisation of several arylaliphatic carboxylic acids. By saturation–transfer–difference NMR spectroscopy (STD-NMR), substrate binding during catalysis was investigated. All atoms of the substrate showed interactions with the enzyme. STD-NMR measurements revealed distinct nuclear Overhauser effects in experiments with and without molecular conversion. The spectroscopic analysis led to the identification of several amino acid residues whose variation increased the activity of G74C. While single-amino acid exchanges increased the activity moderately, structure-guided saturation mutagenesis yielded a quadruple mutant with a 40 times higher reaction rate. This study presents STD-NMR as versatile tool for the analysis of enzyme-substrate interactions in catalytically competent systems and for the guidance of protein engineering.

Keywords: racemase, rational protein design, STD-NMR, structure guided saturation mutagenesis

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Authors: Tassia Batista Pessato, Francielli Pires Ribeiro Morais, Fernanda Guimaraes Drummond Silva, Flavia Maria Netto

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Proteins and phenolic compounds can interact mainly by hydrophobic interactions. Those interactions may lead to structural changes in both molecules, which in turn could affect positively or negatively their functional and nutritional properties. Here, the structural changes of whey proteins (WPI) due to interaction with caffeic acid (CA) were investigated by intrinsic and extrinsic fluorescence. The effects of protein-phenolic compounds interactions on the total phenolic content and antioxidant activity were also assessed. The WPI-CA complexes were obtained by mixture of WPI and CA stock solutions in deionized water. The complexation was carried out at room temperature during 60 min, using 0.1 M NaOH to adjust pH at 7.0. The WPI concentration was fixed at 5 mg/mL, whereas the CA concentration varied in order to obtain four different WPI:CA molar relations (1:1; 2:1; 5:1; 10:1). WPI and phenolic solutions were used as controls. Intrinsic fluorescence spectra of the complexes (mainly due to Trp fluorescence emission) were obtained at λex = 280 nm and the emission intensities were measured from 290 to 500 nm. Extrinsic fluorescence was obtained as the measure of protein surface hydrophobicity (S0) using ANS as a fluorescence probe. Total phenolic content was determined by Folin-Ciocalteau and the antioxidant activity by FRAP and ORAC methods. Increasing concentrations of CA resulted in decreasing of WPI intrinsic fluorescence. The emission band of WPI red shifted from 332 to 354 nm as the phenolic concentration increased, which is related to the exposure of Trp residue to the more hydrophilic environment and unfolding of protein structure. In general, the complexes presented lower S0 values than WPI, suggesting that CA hindered ANS binding to hydrophobic sites of WPI. The total phenolic content in the complexes was lower than the sum of two compounds isolated. WPI showed negligible AA measured by FRAP. However, as the relative concentration of CA increased in the complexes, the FRAP values enhanced, indicating that AA measure by this technique comes mainly from CA. In contrast, the WPI ORAC value (82.3 ± 1.5 µM TE/g) suggest that its AA is related to the capacity of H+ transfer. The complexes exhibited no important improvement of AA measured by ORAC in relation to the isolated components, suggesting complexation partially suppressed AA of the compounds. The results hereby presented indicate that interaction of WPI and CA occurred, and this interaction caused a structural change in the proteins. The complexation can either hide or expose antioxidant sites of both components. In conclusion, although the CA can undergo an AA suppression due to the interaction with proteins, the AA of WPI could be enhanced due to protein unfolding and exposure of antioxidant sites.

Keywords: bioactive properties, milk proteins, phenolic acids, protein-phenolic compounds complexation

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Authors: Adil Basuki Ahza, Michael Liong, Subarna Suryatman

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Healthy chips now dominating the snack market shelves. More than 80% processed snack foods in the market are chips. This research takes the advantages of twin extrusion technology to produce two types of product, i.e. directly expanded and intermediate ready-to-fry or microwavable chips. To improve the functional quality, the cereal-tuber based mix was enriched with antioxidant rich mix of temurui, celery, carrot and isolated soy protein (ISP) powder. Objectives of this research were to find best composite cassava-sorghum ratio, i.e. 60:40, 70:30 and 80:20, to optimize processing conditions of extrusion and study the microstructural, physical and sensorial characteristics of the final products. Optimization was firstly done by applying metering section of extruder barrel temperatures of 120, 130 and 140 °C with screw speeds of 150, 160 and 170 rpm to produce direct expanded product. The intermediate product was extruded in 100 °C and 100 rpm screw speed with feed moisture content of 35, 40 and 45%. The directly expanded products were analyzed for color, hardness, density, microstructure, and organoleptic properties. The results showed that interaction of ratio of cassava-sorghum and cooking methods affected the product's color, hardness, and bulk density (p<0.05). Extrusion processing conditions also significantly affected product's microstructure (p<0.05). The direct expanded snacks of 80:20 cassava-sorghum ratio and fried expanded one 70:30 and 80:20 ratio shown the best organoleptic score (slightly liked) while baking the intermediate product with microwave were resulted sensorial not acceptable quality chips.

Keywords: cassava-sorghum composite, extrusion, microstructure, physical characteristics

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Authors: Reham H. Soliman, Noha Noufal, Howayda AbdelAal

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Calcium/calmodulin-dependent serine protein kinase (CASK) belongs to the membrane-associated guanylate kinase (MAGUK) family and has been proposed as a mediator of cell-cell adhesion and proliferation, which can contribute to tumorogenesis. CASK has been linked as a good prognostic factor with some tumor subtypes, while considered as a poor prognostic marker in others. To our knowledge, no sufficient evidence of CASK role in colorectal cancer is available. The aim of this study is to evaluate the expression of Calcium/calmodulin-dependent serine protein kinase (CASK) in non-mucinous colorectal adenocarcinoma and adenomatous polyps as precursor lesions and assess its prognostic significance. The study included 42 cases of conventional colorectal adenocarcinoma and 15 biopsies of adenomatous polyps with variable degrees of dysplasia. They were reviewed for clinicopathological prognostic factors and stained by CASK; mouse, monoclonal antibody using heat-induced antigen retrieval immunohistochemical techniques. The results showed that CASK protein was significantly overexpressed (p <0.05) in CRC compared with adenoma samples. The CASK protein was overexpressed in the majority of CRC samples with 85.7% of cases showing moderate to strong expression, while 46.7% of adenomas were positive. CASK overexpression was significantly correlated with both TNM stage and grade of differentiation (p <0.05). There was a significantly higher expression in tumor samples with early stages (I/II) rather than advanced stage (III/IV) and with low grade (59.5%) rather than high grade (40.5%). Another interesting finding was found among the adenomas group, where the stronger intensity of staining was observed in samples with high grade dysplasia (33.3%) than those of lower grades (13.3%). In conclusion, this study shows that there is significant overexpression of CASK protein in CRC as well as in adenomas with high grade dysplasia. This indicates that CASK is involved in the process of carcinogenesis and functions as a potential trigger of the adenoma-carcinoma cascade. CASK was significantly overexpressed in early stage and low-grade tumors rather than tumors with advanced stage and higher histological grades. This suggests that CASK protein is a good prognostic factor. We suggest that CASK affects CRC in two different ways derived from its physiology. CASK as part of MAGUK family can stimulate proliferation and through its cell membrane localization and as a mediator of cell-cell adhesion might contribute in tumor confinement and localization.

Keywords: CASK, colorectal cancer, overexpression, prognosis

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Authors: J. O. Idoko, C. N. Michael, T. O. Fasuan

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This study investigated the effects of cooking time, seed-to-water ratio and soaking time on proximate and functional properties of African walnut seed using Box-Behnken design and Response Surface Methodology (BBD-RSM) with a view to increase its utilization in the food industry. African walnut seeds were sorted washed, soaked, cooked, dehulled, sliced, dried and milled. Proximate analysis and functional properties of the samples were evaluated using standard procedures. Data obtained were analyzed using descriptive and inferential statistics. Quadratic models were obtained to predict the proximate and functional qualities as a function of cooking time, seed-to-water ratio and soaking time. The results showed that the crude protein ranged between 11.80% and 23.50%, moisture content ranged between 1.00% and 4.66%, ash content ranged between 3.35% and 5.25%, crude fibre ranged from 0.10% to 7.25% and carbohydrate ranged from 1.22% to 29.35%. The functional properties showed that soluble protein ranged from 16.26% to 42.96%, viscosity ranged from 23.43 mPas to 57 mPas, emulsifying capacity ranged from 17.14% to 39.43% and water absorption capacity ranged from 232% to 297%. An increase in the volume of water used during cooking resulted in loss of water soluble protein through leaching, the length of soaking time and the moisture content of the dried product are inversely related, ash content is inversely related to the cooking time and amount of water used, extraction of fat is enhanced by increase in soaking time while increase in cooking and soaking times result into decrease in fibre content. The results obtained indicated that African walnut could be used in several food formulations as protein supplement and binder.

Keywords: African walnut, functional properties, proximate analysis, response surface methodology

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Authors: Nurul-Akidah M. Yusak, Rahmah Mohamed, Noor Zuhaira Abd Aziz

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The introduction of degradable plastic materials into agricultural sectors has represented a promising alternative to promote green agriculture and environmental friendly of modern farming practices. Major challenges of developing degradable agricultural films are to identify the most feasible types of degradation mechanisms, composition of degradable polymers and related processing techniques. The incorrect choice of degradable mechanisms to be applied during the degradation process will cause premature losses of mechanical performance and strength. In order to achieve controlled process of agricultural film degradation, the compositions of degradable agricultural film also important in order to stimulate degradation reaction at required interval of time and to achieve sustainability of the modern agricultural practices. A set of photodegradable polyethylene based agricultural film was developed and produced, following the selective optimization of processing parameters of the agricultural film manufacturing system. Example of agricultural films application for oil palm seedlings cultivation is presented.

Keywords: photodegradable polyethylene, plasticulture, processing schemes

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Authors: Pedro Henrique Alves Martins, Paulo Guilherme Ferreira De Siqueira, Franco De Castro Bubani, Maria Teresa Paulino Aguilar, Paulo Roberto Cetlin

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Cu-base shape-memory alloys (CuZnAl, CuAlNi, CuAlBe, etc.) are promising engineering materials for several unconventional devices, such as sensors, actuators, and mechanical vibration dampers. Brittleness is one of the factors that limit the commercial use of these alloys, as it makes thermomechanical processing difficult. In this work, a method for the hot extrusion of a 75.50% Cu, 16,74% Zn, 7,76% Al (weight %) alloy is presented. The effects of the thermomechanical processing in the microstructure and the pseudoelastic behavior of the alloy are assessed by optical metallography, compression and hardness tests. Results show that hot extrusion is a suitable method to obtain severe cross-section reductions in the CuZnAl shape-memory alloy studied. The alloy maintained its pseudoelastic effect after the extrusion and the modifications in the mechanical behavior caused by precipitation during hot extrusion can be minimized by a suitable precipitate dissolution heat treatment.

Keywords: hot extrusion, pseudoelastic, shape-memory alloy, thermomechanical processing

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Authors: Rachel Matar, Maxime Merheb

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Chemical drugs have been used for many centuries as the only way to cure diseases until the novel gene therapy has been created in 1960. Gene therapy is based on the insertion, correction, or inactivation of genes to treat people with genetic illness (1). Gene therapy has made wonders in Parkison’s, Alzheimer and multiple sclerosis. In addition to great promises in the healing of deadly diseases like many types of cancer and autoimmune diseases (2). This method implies the use of recombinant DNA technology with the help of different viral and non-viral vectors (3). It is nowadays used in somatic cells as well as embryos and gametes. Beside all the benefits of gene therapy, this technique is deemed by some opponents as an ethically unacceptable treatment as it implies playing with the genes of living organisms.

Keywords: gene therapy, genetic disease, cancer, multiple sclerosis

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Authors: Lai Yi-Hsiu

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The present paper aims to examine the language processing of Chinese-speaking seniors with Alzheimer’s disease (AD) from the perspective of temporal cues. Twenty healthy adults, 17 healthy seniors, and 13 seniors with AD in Taiwan participated in this study to tell stories based on two sets of pictures. Nine temporal cues were fetched and analyzed. Oral productions in Mandarin Chinese were compared and discussed to examine to what extent and in what way these three groups of participants performed with significant differences. Results indicated that the age effects were significant in filled pauses. The dementia effects were significant in mean duration of pauses, empty pauses, filled pauses, lexical pauses, normalized mean duration of filled pauses and lexical pauses. The findings reported in the current paper help characterize the nature of language processing in seniors with or without AD, and contribute to the interactions between the AD neural mechanism and their temporal parameters.

Keywords: language processing, Alzheimer’s disease, Mandarin Chinese, temporal cues

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Authors: Gulyas Erzsebet

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The multimerisation of proteins is a common feature of many cellular processes; however, it could also impair protein functions and/or be associated with the occurrence of diseases. Thus, development of a research tool monitoring the appearance/presence of multimeric protein forms has great importance for a variety of research fields. Such a tool is potentially applicable in the ante-mortem diagnosis of certain conformational diseases, such as transmissible spongiform encephalopathies (TSE) and Alzheimer’s disease. These conditions are accompanied by the appearance of aggregated protein multimers, present in low concentrations in various tissues. This detection is particularly relevant for TSE where the handling of tissues derived from affected individuals and of meat products of infected animals have become an enormous health concern. Here we demonstrate the potential of such a multimer detection approach in TSE by developing a facile approach. The Biospiral-Detect system resembles a traditional sandwich ELISA, except that the capturing antibody that is attached to a solid surface and the detecting antibody is directed against the same or overlapping epitopes. As a consequence, the capturing antibody shields the epitope on the captured monomer from reacting with the detecting antibody, therefore monomers are not detected. Thus, MDS is capable of detecting only protein multimers with high specificity. We developed an alternative system as well, where RNA aptamers were employed instead of monoclonal antibodies. In order to minimize degradation, the 3' and 5' ends of the aptamer contained deoxyribonucleotides and phosphorothioate linkages. When compared the monoclonal antibodies-based system with the aptamers-based one, the former proved to be superior. Thus all subsequent experiments were conducted by employing the Biospiral -Detect modified sandwich ELISA kit. Our approach showed an order of magnitude higher sensitivity toward mulimers than monomers suggesting that this approach may become a valuable diagnostic tool for conformational diseases that are accompanied by multimerization.

Keywords: diagnosis, ELISA, Prion, TSE

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Authors: R. H. Bustos, L. Martinez, J. García, F. Suárez

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MRP1 (Multi-drug resistance associated protein 1) and MRP2 (Multi-drug resistance associated protein 2) are two proteins belonging to the transporters of ABC (ATP-Binding Cassette). These transporter proteins are involved in the efflux of several biological drugs and xenobiotic and also in multiple physiological, pathological and pharmacological processes. Evidence has been found that there is a correlation among different polymorphisms found and their clinical implication in the resistance to antiepileptic, chemotherapy and anti-infectious drugs. In our study, exonic regions of MRP1/ABCC1 y MRP2/ABCC2 were studied in the Colombian population, specifically in the region of the central Savannah (Cundinamarca) to determinate SNP (Single Nucleotide Polymorphisms) and determinate its allele frequency and its genomics frequency. Results showed that for our population, SNP are found that have been previously reported for MRP1/ABCC1 (rs200647436, rs200624910, rs150214567) as well as for MRP2/ABCC2 (rs2273697, rs3740066, rs142573385, rs17216212). In addition, 13 new SNP were identified. Evidences show an important clinic correlation for polymorphisms rs3740066 and rs2273697. The study object population displays genetic variability as compared to the one reported in other populations.

Keywords: ATP-binding cassette (ABCC), Colombian population, multidrug-resistance protein (MRP), pharmacogenetic, single nucleotide polymorphism (SNP)

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Authors: Sahil Kumar, Rajaram Ghadge, Ramesh Bhujade

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Historically, lipid extraction from dried algal biomass remained a focus area of the algal research. It has been realized over the past few years that the lipid-centric approach and conversion technologies that require dry algal biomass have several challenges. Algal culture in cultivation systems contains more than 99% water, with algal concentrations of just a few hundred milligrams per liter ( < 0.05 wt%), which makes harvesting and drying energy intensive. Drying the algal biomass followed by extraction also entails the loss of water and nutrients. In view of these challenges, focus has shifted toward developing processes that will enable oil production from wet algal biomass without drying. Hydrothermal liquefaction (HTL), an emerging technology, is a thermo-chemical conversion process that converts wet biomass to oil and gas using water as a solvent at high temperature and high pressure. HTL processes wet algal slurry containing more than 80% water and significantly reduces the adverse cost impact owing to drying the algal biomass. HTL, being inherently feedstock agnostic, i.e., can convert carbohydrates and proteins also to fuels and recovers water and nutrients. It is most effective with low-lipid (10--30%) algal biomass, and bio-crude yield is two to four times higher than the lipid content in the feedstock. In the early 2010s, research remained focused on increasing the oil yield by optimizing the process conditions of HTL. However, various techno-economic studies showed that simply converting algal biomass to only oil does not make economic sense, particularly in view of low crude oil prices. Making the best use of every component of algae is a key for economic viability of algal to oil process. On investigation of HTL reactions at the molecular level, it has been observed that sequential HTL has the potential to recover value-added products along with biocrude and improve the overall economics of the process. This potential of sequential HTL makes it a most promising technology for converting wet waste to wealth. In this presentation, we will share our experience on the techno-economic and engineering aspects of sequential HTL for conversion of algal biomass to algal bio-oil and co-products.

Keywords: algae, biomass, lipid, protein

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Authors: Hee-Choul Kwon, Hyungjin Cho, Heeyong Kwon

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Image rotation is one of main pre-processing step in image processing or image pattern recognition. It is implemented with rotation matrix multiplication. However it requires lots of floating point arithmetic operations and trigonometric function calculations, so it takes long execution time. We propose a new high speed image rotation algorithm without two major time-consuming operations. We compare the proposed algorithm with the conventional rotation one with various size images. Experimental results show that the proposed algorithm is superior to the conventional rotation ones.

Keywords: high speed rotation operation, image processing, image rotation, pattern recognition, transformation matrix

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Authors: Retno Supriyanti, Teguh Budiono, Yogi Ramadhani, Haris B. Widodo, Arwita Mulyawati

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Cervical cancer is the leading cause of mortality in cancer-related diseases. In this diagnosis doctors usually perform several tests to determine the presence of cervical cancer in a patient. However, these checks require support equipment to get the results in more detail. One is by using ultrasonography. However, for the developing countries most of the existing ultrasonography has a low resolution. The goal of this research is to obtain abnormalities on low-resolution ultrasound images especially for cervical cancer case. In this paper, we emphasize our work to use Image Enhancement for pre-processing image quality improvement. The result shows that pre-processing stage is promising to support further analysis.

Keywords: cervical cancer, mortality, low-resolution, image enhancement.

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Authors: Ana Cleide Vieira Gomes Guimbal de Aquino

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Sound communication is present in most vertebrates, from fish, mainly in species that live in murky waters, to some species of reptiles, anuran amphibians, birds, and mammals, including primates. There are, in fact, relevant similarities between human language and animal sound communication, and among these similarities are the vocalizations called calls. The first specific call in human babies is crying, which has a characteristic prosodic contour and is motivated most of the time by the need for food and by affecting the puppy-caregiver interaction, with a view to communicating the necessities and food requests and guaranteeing the survival of the species. The present work aims to articulate speech processing in the motor context with aspects of the project entitled emotional states and vocalization: a comparative study of the prosodic contours of crying in human and non-human animals. First, concepts of speech motor processing and general aspects of speech evolution will be presented to relate these two approaches to animal sound communication.

Keywords: speech motor processing, animal communication, animal behaviour, language acquisition

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Authors: Syeda Fahria Hoque Mimmi

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Many new and promising treatments for reducing or diminishing the adverse effects of microorganisms are being discovered day by day. On the other hand, the dairy industry is accelerating the economic wheel of Bangladesh. Considering all these facts, new thoughts were developed to isolate milk proteins by the present experiment for opening up a new era of developing natural antibiotics from milk. Lactoferrin, an iron-binding glycoprotein with multifunctional properties, is crucial to strengthening the immune system and also useful for commercial applications. The protein’s iron-binding capacity makes it undoubtedly advantageous to immune system modulation and different bacterial strains. For fulfilling the purpose, 4 of raw and 17 of commercially available milk samples were collected from different farms and stores in Bangladesh (Dhaka, Chittagong, and Cox’s Bazar). Protein quantification by nanodrop technology has confirmed that raw milk samples have better quantities of protein than the commercial ones. All the samples were tested for their antimicrobial activity against 18 pathogens, where raw milk samples showed a higher percentage of antibacterial activity. In addition to this, SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis) was performed to identify lactoferrin in the milk samples. Lactoferrin was detected in 9 samples from which 4 were raw milk samples. Interestingly, Streptococcus pyogenes, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, Vibrio cholera, Staphylococcus aureus, and enterotoxigenic E. coli significantly displayed sensitivity against lactoferrin collected from raw milk. Only Bacillus cereus, Pseudomonas aeruginosa, Streptococcus pneumonia, Enterococcus faecalis, and ETEC (Enterotoxigenic Escherichia coli) were susceptible to lactoferrin obtained from a commercial one. This study suggested that lactoferrin might be used as the potential alternative of antibiotics for many diseases and also can be used to reduce microbial deterioration in the food and feed industry.

Keywords: alternative of antibiotics, commercially available milk, lactoferrin, nanodrop technology, pathogens, raw milk

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Authors: H. M. Soroush

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The problem of scheduling products and services for on-time deliveries is of paramount importance in today’s competitive environments. It arises in many manufacturing and service organizations where it is desirable to complete jobs (products or services) with different weights (penalties) on or before their due dates. In such environments, schedules should frequently decide whether to schedule a job based on its processing time, due-date, and the penalty for tardy delivery to improve the system performance. For example, it is common to measure the weighted number of late jobs or the percentage of on-time shipments to evaluate the performance of a semiconductor production facility or an automobile assembly line. In this paper, we address the problem of scheduling a set of jobs on a server where processing times or due-dates of jobs are random variables and fixed weights (penalties) are imposed on the jobs’ late deliveries. The goal is to find the schedule that minimizes the expected weighted number of tardy jobs. The problem is NP-hard to solve; however, we explore three scenarios of the problem wherein: (i) both processing times and due-dates are stochastic; (ii) processing times are stochastic and due-dates are deterministic; and (iii) processing times are deterministic and due-dates are stochastic. We prove that special cases of these scenarios are solvable optimally in polynomial time, and introduce efficient heuristic methods for the general cases. Our computational results show that the heuristics perform well in yielding either optimal or near optimal sequences. The results also demonstrate that the stochasticity of processing times or due-dates can affect scheduling decisions. Moreover, the proposed problem is general in the sense that its special cases reduce to some new and some classical stochastic single machine models.

Keywords: number of late jobs, scheduling, single server, stochastic

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Authors: Atlal El-Assaad

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Viruses change with time through mutations and result in new variants that may persist or disappear. A Mutation refers to an actual change in the virus genetic sequence, and a variant is a viral genome that may contain one or more mutations. Critical mutations may cause the virus to be more transmissible, with high disease severity, and more vulnerable to diagnostics, therapeutics, and vaccines. Thus, variants carrying such mutations may increase the risk to human health and are considered variants of concern (VOC). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) - the contagious in humans, positive-sense single-stranded RNA virus that caused coronavirus disease 2019 (COVID-19) - has been studied thoroughly, and several variants were revealed across the world with their corresponding mutations. SARS-CoV-2 has four structural proteins, known as the S (spike), E (envelope), M (membrane), and N (nucleocapsid) proteins, but prior study and vaccines development focused on genetic mutations in the S protein due to its vital role in allowing the virus to attach and fuse with the membrane of a host cell. Specifically, subunit S1 catalyzes attachment, whereas subunit S2 mediates fusion. In this perspective, we studied all charged amino acid mutations of the SARS-CoV-2 viral spike protein S1 when bound to Antibody CC12.1 in a crystal structure and assessed the effect of different mutations. We generated all missense mutants of SARS-CoV-2 protein amino acids (AAs) within the SARS-CoV-2:CC12.1 complex model. To generate the family of mutants in each complex, we mutated every charged amino acid with all other charged amino acids (Lysine (K), Arginine (R), Glutamic Acid (E), and Aspartic Acid (D)) and studied the new binding of the complex after each mutation. We applied Poisson-Boltzmann electrostatic calculations feeding into free energy calculations to determine the effect of each mutation on binding. After analyzing our data, we identified charged amino acids keys for binding. Furthermore, we validated those findings against published experimental genetic data. Our results are the first to propose in silico potential life-threatening mutations of SARS-CoV-2 beyond the present mutations found in the five common variants found worldwide.

Keywords: SARS-CoV-2, variant, ionic amino acid, protein-protein interactions, missense mutation, AESOP

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Authors: Sri Satya Antarlina, Elok Zubaidah, Teti Istiana, Harijono

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Sorghum bicolor (Sorghum bicolor L. Moench) has the potential as a flour for gluten-free food products. Sorghum flour production needs grain soaking treatment. Soaking can reduce the tannin content which is an anti-nutrient, so it can increase the protein digestibility. Fine particle size decreases the yield of flour, so it is necessary to study various particle sizes to increase the yield. This study aims to determine the characteristics of sorghum flour in the treatment of soaking peeled grain and particle size. The material of white sorghum varieties KD-4 from farmers in East Java, Indonesia. Factorial randomized factorial design (two factors), repeated three times, factor I were the time of grain soaking (five levels) that were 0, 12, 24, 36, and 48 hours, factor II was the size of the starch particles sifted with a fineness level of 40, 60, 80, and 100 mesh. The method of making sorghum flour is grain peeling, soaking peeled grain, drying using the oven at 60ᵒC, milling, and sieving. Physico-chemical analysis of sorghum flour. The results show that there is an interaction between soaking time of grain with the size of sorghum flour particles. Interaction in yield of flour, L* color (brightness level), whiteness index, paste properties, amylose content, protein content, bulk density, and protein digestibility. The method of making sorghum flour through the soaking of peeled grain and the difference in particle size has an important role in producing the physicochemical properties of the specific flour. Based on the characteristics of sorghum flour produced, it is determined the method of making sorghum flour through sorghum grain soaking for 24 hours, the particle size of flour 80 mesh. The sorghum flour with characteristic were 24.88% yield of flour, 88.60 color L* (brightness level), 69.95 whiteness index, 3615 Cp viscosity, 584.10 g/l of bulk density, 24.27% db protein digestibility, 90.02% db starch content, 23.4% db amylose content, 67.45% db amylopectin content, 0.22% db crude fiber content, 0.037% db tannin content, 5.30% db protein content, ash content 0.18% db, carbohydrate content 92.88 % db, and 1.94% db fat content. The sorghum flour is recommended for cookies products.

Keywords: characteristic, sorghum (Sorghum bicolor L. Moench) flour, grain soaking, particle size, physicochemical properties

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Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

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High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

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Authors: Z. O. Apotiola, J. F. Fashakin

Abstract:

This project studied the production and evaluation of jam produced from pineapple and grape at different level of ratio (90:10, 80:20, 70:30, 60:40, 50:50, and 100%). The proximate and sensory properties were determined using standard methods. The (GDZ) was the highest for protein, moisture, fat and ash, (KFJ) was the highest for carbohydrate. There were significant differences (p<0.05) in samples (PAB, GDZ, BEN) for moisture. Also, there were significant differences (p<0.05) in samples (PAB, BBL, GDZ, KFJ) for protein. There were significant differences (p<0.05) in samples (PAB, BBL, BEN) for carbohydrate. Also, there were significant differences (p<0.05) in samples (PAB, BBL, QCM, GDZ, BEN) for fat and there were significant differences (p<0.05) in samples (PAB, BBL, GDZ) for ash. (KFJ) was the highest for pH, (BBL and QCM) was the highest for Vitamin C; (GDZ) was the highest for titratable acidity. For sensory properties, for aroma, colour, flavour, and overall acceptability were tested using panellists; the result showed that (KFJ) had the highest for all samples. From the results of chemical and sensory characteristics sample BBL was the best combination.

Keywords: chemical, characteristic, combination, titratable, sensory, significant

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Authors: K. R. Thabethe, G. A. Adefolaju, M. J. Hosie

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Cervical cancer is the third most commonly diagnosed cancer globally and it is one of three AIDS defining malignancies. Highly active antiretroviral therapy (HAART) is a combination of three or more antiretroviral drugs and has been shown to play a significant role in reducing the incidence of some AIDS defining malignancies, although its effect on cervical cancer is still unclear. The aim of this study was to investigate the relationship between cervical cancer and HAART. This was achieved by studying the expression of two signalling molecules expressed in cervical cancer; MUC1 and P65. Following the 24 hour treatment of a cervical cancer cell line, HCS-2, with drugs which are commonly used as part of HAART at their clinical plasma concentrations, real-time qPCR and immunofluorescence were used in order to study gene and protein expression. A one way ANOVA followed by a Tukey Kramer Post Hoc test was conducted using JMP 11 software on both sets of data. The drug classified as a protease inhibitor (PI) (i.e. LPV/r) reduced MUC1 and P65 gene and protein expression more than the other drug tested. PIs are known to play a significant role in cell death, therefore the cells were thought to be more susceptible to cell death following treatment with PIs. In conclusion, the drugs used, especially the PI showed some anticancer effects by facilitating cell death through decreased gene and protein expression of MUC1 and P65 and present promising agents for cancer treatment.

Keywords: cervical cancer, haart, MUC1, P65

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Authors: V. Singh, S. Singh, S. S. Garewal

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Metal matrix composites with aluminum as the matrix material have been heralded as the next great development in advanced engineering materials. Aluminum metal matrix composites (AMMC) refer to the class of light weight high performance material systems. Properties of AMMCs can be tailored to the demands of different industrial applications by suitable combinations of matrix, reinforcement and processing route. AMMC finds its application in automotive, aerospace, defense, sports and structural areas. This paper presents an overview of AMMC material systems on aspects relating to processing, types and applications with case studies.

Keywords: aluminum metal matrix composites, applications of aluminum metal matrix composites, lighting material processing of aluminum metal matrix composites

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Authors: Fatemeh Sadat Miri, Morteza Ehsani, Seyed Farshid Hosseini

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The present research is concerned with the optimization of extrusion parameters of ABS sheets by the Taguchi experimental design method. In this design method, three parameters of % recycling ABS, processing temperature and degassing time on mechanical properties, hardness, HDT, and color matching of ABS sheets were investigated. The variations of this research are the dosage of recycling ABS, processing temperature, and degassing time. According to experimental test data, the highest level of tensile strength and HDT belongs to the sample with 5% recycling ABS, processing temperature of 230°C, and degassing time of 3 hours. Additionally, the minimum level of MFI and color matching belongs to this sample, too. The present results are in good agreement with the Taguchi method. Based on the outcomes of the Taguchi design method, degassing time has the most effect on the mechanical properties of ABS sheets.

Keywords: ABS, process optimization, Taguchi, mechanical properties

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Authors: Sinethemba Yakobi, Lindiwe Zuma, Ofentse Pooe

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Gonococcal infections present a notable public health issue, and the major approach for treatment involves using β-lactam antibiotics that specifically target penicillin-binding protein 2 (PBP2) in Neisseria gonorrhoeae. This study examines the influence of flavonoids, namely rutin, on the structural changes of PBP2 in both penicillin-resistant (FA6140) and penicillin-susceptible (FA19) strains. The research clarifies the structural effects of particular mutations, such as inserting an aspartate residue at position 345 (Asp-345a) in the PBP2 protein. The strain FA6140, which is resistant to penicillin, shows specific changes that lead to a decrease in penicillin binding. These mutations, namely P551S and F504L, significantly impact the pace at which acylation occurs and the stability of the strain under high temperatures. Molecular docking analyses investigate the antibacterial activities of rutin and other phytocompounds, emphasizing its exceptional binding affinity and potential as an inhibitor of PBP2. Quercetin and protocatechuic acid have encouraging antibacterial effectiveness, with quercetin displaying characteristics similar to those of drugs. Molecular dynamics simulations offer a detailed comprehension of the interactions between flavonoids and PBP2, highlighting rutin's exceptional antioxidant effects and strong affinity for the substrate binding site. The study's wider ramifications pertain to the pressing requirement for antiviral treatments in the context of the ongoing COVID-19 epidemic. Flavonoids have a strong affinity for binding to PBP2, indicating their potential as inhibitors to impair cell wall formation in N. gonorrhoeae. Ultimately, this study provides extensive knowledge on the interactions between proteins and ligands, the dynamics of the structure, and the ability of flavonoids to combat penicillin-resistant N. gonorrhoeae bacteria. The verified simulation outcomes establish a basis for creating potent inhibitors and medicinal therapies to combat infectious illnesses.

Keywords: phytochemicals, penicillin-binding protein 2, gonococcal infection, ligand-protein interaction, binding energy, neisseria gonorrhoeae FA19, neisseria gonorrhoeae FA6140, flavonoids

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Authors: Mirnawati, G. Ciptaan, Ferawati

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This study aims to improve the quality of the cassava peel-leaf mixture (CPLM) through fermentation with Rhizopus oligosporusas poultry ration. This research is an experimental study using a completely randomized design (CRD) with four treatments and five replications. The treatments were cassava peel-leaf mixture (CPLM) fermented with Rhizopus oligosporus. The treatments were a combination of cassava peel and leaves with the ratio of; A (9:1), B (8:2), C (7:3), and D (6:4). The observed variables were protease enzyme activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and metabolic energy. The results of the diversity analysis showed that there was a very significant (p < 0.01) effect on protease activity, crude protein, crude fiber, nitrogen retention, digestibility of crude fiber, and energy metabolism of fermented CPLM. Based on the results of the study, it can be concluded that CPLM (6:4) fermented with Rhizopus oligosporus gave the best results seen from protease activity 7,25 U/ml, 21.23% crude protein, 19.80% crude fiber, 59.65% nitrogen retention, 62.99% crude fiber digestibility and metabolic energy 2671 Kcal/kg.

Keywords: quality, Casava peel-leaf mixture, fermentation, Rhizopus oligosporus

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Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

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Authors: Diana Pombo, Maria Loaiza, Mauricio Quijano, Alberto Cadena, Juan Pablo Tello

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In this paper, we present a semi-automatic tool for calculating ejection fraction from an echocardiographic video signal which is derived from a database in DICOM format, of Clinica de la Costa - Barranquilla. Described in this paper are each of the steps and methods used to find the respective calculation that includes acquisition and formation of the test samples, processing and finally the calculation of the parameters to obtain the ejection fraction. Two imaging segmentation methods were compared following a methodological framework that is similar only in the initial stages of processing (process of filtering and image enhancement) and differ in the end when algorithms are implemented (Active Contour and Region Growing Algorithms). The results were compared with the measurements obtained by two different medical specialists in cardiology who calculated the ejection fraction of the study samples using the traditional method, which consists of drawing the region of interest directly from the computer using echocardiography equipment and a simple equation to calculate the desired value. The results showed that if the quality of video samples are good (i.e., after the pre-processing there is evidence of an improvement in the contrast), the values provided by the tool are substantially close to those reported by physicians; also the correlation between physicians does not vary significantly.

Keywords: echocardiography, DICOM, processing, segmentation, EDV, ESV, ejection fraction

Procedia PDF Downloads 426