Search results for: rare sensitive attribute
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2952

Search results for: rare sensitive attribute

1122 Bacteriological Culture Methods and its Uses in Clinical Pathology

Authors: Prachi Choudhary, Jai Gopal Sharma

Abstract:

Microbial cultures determine the type of organism, its abundance in the tested sample, or both. It is one of the primary diagnostic methods of microbiology. It is used to determine the cause of infectious disease by letting the agent multiply in a predetermined medium. Different bacteria produce colonies that may be very distinct from the bacterial species that produced them. To culture any pathogen or microorganism, we should first know about the types of media used in microbiology for culturing. Sometimes sub culturing is also done in various microorganisms if some mixed growth is seen in culture. Nearly 3 types of culture media based on consistency – solid, semi-solid, and liquid (broth) media; are further explained in the report. Then, The Five I's approach is a method for locating, growing, observing, and characterizing microorganisms, including inoculation and incubation. Isolation, inspection, and identification. For identification of bacteria, we have to culture the sample like urine, sputum, blood, etc., on suitable media; there are different methods of culturing the bacteria or microbe like pour plate method, streak plate method, swabbing by needle, pipetting, inoculation by loop, spreading by spreader, etc. After this, we see the bacterial growth after incubation of 24 hours, then according to the growth of bacteria antibiotics susceptibility test is conducted; this is done for sensitive antibiotics or resistance to that bacteria, and also for knowing the name of bacteria. Various methods like the dilution method, disk diffusion method, E test, etc., do antibiotics susceptibility tests. After that, various medicines are provided to the patients according to antibiotic sensitivity and resistance.

Keywords: inoculation, incubation, isolation, antibiotics suspectibility test, characterizing

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1121 Impact of Tourism on Sustainability on Essaouira Destination in Morocco

Authors: Hadach Mohamed

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Tourism becomes more and more a source of added value for developing countries. In Morocco, the sector contributes at 20% of national GDP, or the effects of this activity become increasingly harmful. The methodology we followed is qualitative, we analyzed the data according to a process-based approach in two longitudinal period from 2001 to 2009 and a period of real time from 2010 to 2014.Through a process-based longitudinal study we analyzed the effects of tourism on the three components of sustainability: economic, environmental and socio-cultural in Essaouira destination in the south west of Morocco. The objective of this paper is to identify among others, harmful effects of mass tourism on fragile destination in terms of load capacity, promotion of youth employment and respect for indigenous traditions. This study also aims to analyze the impact of tourism on the fragile destination, which depends heavily on this activity; it also seeks to test a series of indicators for sustainable development of sensitive areas. Within results, we found that tourism as an activity is very linked to the international situation, tested sustainable development indicators showed us that tourism is environmentally destructive, job creator and changer modes and lives of indigenous. Between the two periods analyzed, the situation becomes more and more vulnerable and the state intervention is becoming more indispensable. We also found that 70% of the population of the destination does not benefit from the income generated by the destination. This raises questions about the fate of the added value of this activity.

Keywords: economic, environmental and socio-cultural, fragile destination, tourism sustainability

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1120 Effect of Intrinsic Point Defects on the Structural and Optical Properties of SnO₂ Thin Films Grown by Ultrasonic Spray Pyrolysis Method

Authors: Fatiha Besahraoui, M'hamed Guezzoul, Kheira Chebbah, M'hamed Bouslama

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SnO₂ thin film is characterized by Atomic Force Microscopy (AFM) and Photoluminescence Spectroscopies. AFM images show a dense surface of columnar grains with a roughness of 78.69 nm. The PL measurements at 7 K reveal the presence of PL peaks centered in IR and visible regions. They are attributed to radiative transitions via oxygen vacancies, Sn interstitials, and dangling bonds. A bands diagram model is presented with the approximate positions of intrinsic point defect levels in SnO₂ thin films. The integrated PL measurements demonstrate the good thermal stability of our sample, which makes it very useful in optoelectronic devices functioning at room temperature. The unusual behavior of the evolution of PL peaks and their full width at half maximum as a function of temperature indicates the thermal sensitivity of the point defects present in the band gap. The shallower energy levels due to dangling bonds and/or oxygen vacancies are more sensitive to the temperature. However, volume defects like Sn interstitials are thermally stable and constitute deep and stable energy levels for excited electrons. Small redshifting of PL peaks is observed with increasing temperature. This behavior is attributed to the reduction of oxygen vacancies.

Keywords: transparent conducting oxide, photoluminescence, intrinsic point defects, semiconductors, oxygen vacancies

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1119 Effects of Effort and Water Quality on Productivity (CPUE) of Hampal (Hampala macrolepidota) Resources in Jatiluhur Dam, West Java

Authors: Ririn Marinasari, S. Pi

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Hampal (Hampala macrolepidota) is one of Citarum river indigenous fishes that still find in Jatiluhur dam. IUCN at 2013 said that hampal listed on redlist species category, this species was rare in Jatiluhur dam. This species more and more decreasing because change of habitats characteristic such as water quality and fishing effort. This study aims to determine and identify the influence of fishing effort and the quality of water on the productivity of fish resources hampal (Hampala macrolepidota) in Jatiluhur. The study was conducted from October to November 2013. Zones of research include lacustrine zone, transition and Riverin. Hampal fish productivity value computed by Hampal’s CPUE values. The results showed that fish MSY hampal obtained from surplus production model of Schaefer is equal to 0.2045 tons / quarterly. In the years 2011-2012 have occurred over fishing in 2013 while still under fishing. Total catches have exceeded the MSY during the year 2011 and the third quarterly of 2012 tons of fish that exceed 0.2045 hampal. The rate of utilization of fish resources hampal is equal to 80% of MSY or equal to the allowable catch (Total Allowable Catch) for fish in Jatiluhur hampal based Schaefer surplus production theory. Fishing effort, water quality parameters such as DO, turbidity and negatively correlated sulfide as H2S, while the temperature and pH positively correlated to productivity or unit catches fish hampal efforts in quarterly time series in the period 2011-2013. Shows that the higher fishing effort, DO, turbidity and sulfide in H2S and diminishing the temperature and pH of the productivity decreases. Variables that affect the productivity of fishing hampal only H2S only factor beta coefficient -0.834 which indicates a negative effect. It can be caused by H2S levels are toxic and have already exceeded the quality standard, while for other water quality parameters are still below the maximum standards allowed in the waters. Result of the study can be a reference of fishing regulation for hampal conservation in Jatiluhur dam.

Keywords: effort, hampal, productivity, water quality

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1118 Targeting Glucocorticoid Receptor Eliminate Dormant Chemoresistant Cancer Stem Cells in Glioblastoma

Authors: Aoxue Yang, Weili Tian, Yonghe Wu, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Little progress has been made in therapeutic treatment of glioblastoma in the last decade despite rapid progress in molecular understanding of brain tumors1. Here we show that the stress hormone glucocorticoid is essential for the maintenance of brain tumor stem cells (BTSCs), which are resistant to conventional therapy. The glucocorticoid receptor (GR) regulates metabolic plasticity and chemoresistance of the dormant BTSC via controlling expression of GPD1 (glycerol-3-phosphate dehydrogenase 1), which is an essential regulator of lipid metabolism in BTSCs. Genomic, lipidomic and cellular analysis confirm that GR/GPD1 regulation is essential for BTSCs metabolic plasticity and survival. We further demonstrate that the GR agonist dexamethasone (DEXA), which is commonly used to control edema in glioblastoma, abolishes the effect of chemotherapy drug temozolomide (TMZ) by upregulating GPD1 and thus promoting tumor cell dormancy in vivo, this provides a mechanistic explanation and thus settle the long-standing debate of usage of steroid in brain tumor patient edema control. Pharmacological inhibition of GR/GPD1 pathway disrupts metabolic plasticity of BTSCs and prolong animal survival, which is superior to standard chemotherapy. Patient case study shows that GR antagonist mifepristone blocks tumor progression and leads to symptomatic improvement. This study identifies an important mechanism regulating cancer stem cell dormancy and provides a new opportunity for glioblastoma treatment.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides.

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1117 Foreign Exchange Volatilities and Stock Prices: Evidence from London Stock Exchange

Authors: Mahdi Karazmodeh, Pooyan Jafari

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One of the most interesting topics in finance is the relation between stock prices and exchange rates. During the past decades different stock markets in different countries have been the subject of study for researches. The volatilities of exchange rates and its effect on stock prices during the past 10 years have continued to be an attractive research topic. The subject of this study is one of the most important indices, FTSE 100. 20 firms with the highest market capitalization in 5 different industries are chosen. Firms are included in oil and gas, mining, pharmaceuticals, banking and food related industries. 5 different criteria have been introduced to evaluate the relationship between stock markets and exchange rates. Return of market portfolio, returns on broad index of Sterling are also introduced. The results state that not all firms are sensitive to changes in exchange rates. Furthermore, a Granger Causality test has been run to observe the route of changes between stock prices and foreign exchange rates. The results are consistent, to some level, with the previous studies. However, since the number of firms is not large, it is suggested that a larger number of firms being used to achieve the best results. However results showed that not all firms are affected by foreign exchange rates changes. After testing Granger Causality, this study found out that in some industries (oil and gas, pharmaceuticals), changes in foreign exchange rate will not cause any changes in stock prices (or vice versa), however, in banking sector the situation was different. This industry showed more reaction to these changes. The results are similar to the ones with Richards and Noel, where a variety of firms in different industries were evaluated.

Keywords: stock prices, foreign exchange rate, exchange rate exposure, Granger Causality

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1116 The Influence of Gender on Job-Competencies Requirements of Chemical-Based Industries and Undergraduate-Competencies Acquisition of Chemists in South West, Nigeria

Authors: Rachael Olatoun Okunuga

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Developing young people’s employability is a key policy issue for ensuring their successful transition to the labour market and their access to career oriented employment. The youths of today irrespective of their gender need to acquire the knowledge, skills and attitudes that will enable them to create or find jobs as well as cope with unpredictable labour market changes throughout their working lives. In a study carried out to determine the influence of gender on job-competencies requirements of chemical-based industries and undergraduate-competencies acquisition by chemists working in the industries, all chemistry graduates working in twenty (20) chemical-based industries that were randomly selected from six sectors of chemical-based industries in Lagos and Ogun States of Nigeria were administered with Job-competencies required and undergraduate-competencies acquired assessment questionnaire. The data were analysed using means and independent sample t-test. The findings revealed that the population of female chemists working in chemical-based industries is low compared with the number of male chemists; furthermore, job-competencies requirements are found not to be gender sensitive while there is no significant difference in undergraduate-competencies acquisition of male and female chemists. This suggests that females should be given the same opportunity of employment in chemical-based industries as their male counterparts. The study also revealed the level of acquisition of undergraduate competencies as related to the needs of chemical-based industries.

Keywords: knowledge, skill, attitude, acquired, required, employability

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1115 Low Field Microwave Absorption and Magnetic Anisotropy in TM Co-Doped ZnO System

Authors: J. Das, T. S. Mahule, V. V. Srinivasu

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Electron spin resonance (ESR) study at 9.45 GHz and a field modulation frequency of 100Hz was performed on bulk polycrystalline samples of Mn:TM (Fe/Ni) and Mn:RE (Gd/Sm) co doped ZnO samples with composition Zn1-xMn:TM/RE)xO synthesised by solid state reaction route and sintered at 500 0C temperature. The room temperature microwave absorption data collected by sweeping the DC magnetic field from -500 to 9500 G for the Mn:Fe and Mn:Ni co doped ZnO samples exhibit a rarely reported non resonant low field absorption (NRLFA) in addition to a strong absorption at around 3350G, usually associated with ferromagnetic resonance (FMR) satisfying Larmor’s relation due to absorption in the full saturation state. Observed low field absorption is distinct to ferromagnetic resonance even at low temperature and shows hysteresis. Interestingly, it shows a phase opposite with respect to the main ESR signal of the samples, which indicates that the low field absorption has a minimum value at zero magnetic field whereas the ESR signal has a maximum value. The major resonance peak as well as the peak corresponding to low field absorption exhibit asymmetric nature indicating magnetic anisotropy in the sample normally associated with intrinsic ferromagnetism. Anisotropy parameter for Mn:Ni codoped ZnO sample is noticed to be quite higher. The g values also support the presence of oxygen vacancies and clusters in the samples. These samples have shown room temperature ferromagnetism in the SQUID measurement. However, in rare earth (RE) co doped samples (Zn1-x (Mn: Gd/Sm)xO), which show paramagnetic behavior at room temperature, the low field microwave signals are not observed. As microwave currents due to itinerary electrons can lead to ohmic losses inside the sample, we speculate that more delocalized 3d electrons contributed from the TM dopants facilitate such microwave currents leading to the loss and hence absorption at the low field which is also supported by the increase in current with increased micro wave power. Besides, since Fe and Ni has intrinsic spin polarization with polarisability of around 45%, doping of Fe and Ni is expected to enhance the spin polarization related effect in ZnO. We emphasize that in this case Fe and Ni doping contribute to polarized current which interacts with the magnetization (spin) vector and get scattered giving rise to the absorption loss.

Keywords: co-doping, electron spin resonance, hysteresis, non-resonant microwave absorption

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1114 Conversion of Glycerol to 3-Hydroxypropanoic Acid by Genetically Engineered Bacillus subtilis

Authors: Aida Kalantari, Boyang Ji, Tao Chen, Ivan Mijakovic

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3-hydroxypropanoic acid (3-HP) is one of the most important biomass-derivable platform chemicals that can be converted into a number of industrially important compounds. There have been several attempts at production of 3-HP from renewable sources in cell factories, focusing mainly on Escherichia coli, Klebsiella pneumoniae, and Saccharomyces cerevisiae. Despite the significant progress made in this field, commercially exploitable large-scale production of 3-HP in microbial strains has still not been achieved. In this study, we investigated the potential of Bacillus subtilis to be used as a microbial platform for bioconversion of glycerol into 3-HP. Our recombinant B. subtilis strains overexpress the two-step heterologous pathway containing glycerol dehydratase and aldehyde dehydrogenase from various backgrounds. The recombinant strains harboring the codon-optimized synthetic pathway from K. pneumoniae produced low levels of 3-HP. Since the enzymes in the heterologous pathway are sensitive to oxygen, we had to perform our experiments in micro-aerobic conditions. Under these conditions, the cell produces lactate in order to regenerate NAD+, and we found the lactate production to be in competition with the production of 3-HP. Therefore, based on the in silico predictions, we knocked out the glycerol kinase (glpk), which in combination with growth on glucose, resulted in improving the 3-HP titer to 1 g/L and the removal of lactate. Cultivation of the same strain in an enriched medium improved the 3-HP titer up to 7.6 g/L. Our findings provide the first report of successful introduction of the biosynthetic pathway for conversion of glycerol into 3-HP in B. subtilis.

Keywords: bacillus subtilis, glycerol, 3-hydroxypropanoic acid, metabolic engineering

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1113 Quantitative Analysis of Caffeine in Pharmaceutical Formulations Using a Cost-Effective Electrochemical Sensor

Authors: Y. T. Gebreslassie, Abrha Tadesse, R. C. Saini, Rishi Pal

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Caffeine, known chemically as 3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-dione, is a naturally occurring alkaloid classified as an N-methyl derivative of xanthine. Given its widespread use in coffee and other caffeine-containing products, it is the most commonly consumed psychoactive substance in everyday human life. This research aimed to develop a cost-effective, sensitive, and easily manufacturable sensor for the detection of caffeine. Antraquinone-modified carbon paste electrode (AQMCPE) was fabricated, and the electrochemical behavior of caffeine on this electrode was investigated using cyclic voltammetry (CV) and square wave voltammetry (SWV) in a solution of 0.1M perchloric acid at pH 0.56. The modified electrode displayed enhanced electrocatalytic activity towards caffeine oxidation, exhibiting a two-fold increase in peak current and an 82 mV shift of the peak potential in the negative direction compared to an unmodified carbon paste electrode (UMCPE). Exploiting the electrocatalytic properties of the modified electrode, SWV was employed for the quantitative determination of caffeine. Under optimized experimental conditions, a linear relationship between peak current and concentration was observed within the range of 2.0 x 10⁻⁶ to 1.0× 10⁻⁴ M, with a correlation coefficient of 0.998 and a detection limit of 1.47× 10⁻⁷ M (signal-to-noise ratio = 3). Finally, the proposed method was successfully applied to the quantitative analysis of caffeine in pharmaceutical formulations, yielding recovery percentages ranging from 95.27% to 106.75%.

Keywords: antraquinone-modified carbon paste electrode, caffeine, detection, electrochemical sensor, quantitative analysis

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1112 Development of Wide Bandgap Semiconductor Based Particle Detector

Authors: Rupa Jeena, Pankaj Chetry, Pradeep Sarin

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The study of fundamental particles and the forces governing them has always remained an attractive field of theoretical study to pursue. With the advancement and development of new technologies and instruments, it is possible now to perform particle physics experiments on a large scale for the validation of theoretical predictions. These experiments are generally carried out in a highly intense beam environment. This, in turn, requires the development of a detector prototype possessing properties like radiation tolerance, thermal stability, and fast timing response. Semiconductors like Silicon, Germanium, Diamond, and Gallium Nitride (GaN) have been widely used for particle detection applications. Silicon and germanium being narrow bandgap semiconductors, require pre-cooling to suppress the effect of noise by thermally generated intrinsic charge carriers. The application of diamond in large-scale experiments is rare owing to its high cost of fabrication, while GaN is one of the most extensively explored potential candidates. But we are aiming to introduce another wide bandgap semiconductor in this active area of research by considering all the requirements. We have made an attempt by utilizing the wide bandgap of rutile Titanium dioxide (TiO2) and other properties to use it for particle detection purposes. The thermal evaporation-oxidation (in PID furnace) technique is used for the deposition of the film, and the Metal Semiconductor Metal (MSM) electrical contacts are made using Titanium+Gold (Ti+Au) (20/80nm). The characterization comprising X-Ray Diffraction (XRD), Atomic Force Microscopy (AFM), Ultraviolet (UV)-Visible spectroscopy, and Laser Raman Spectroscopy (LRS) has been performed on the film to get detailed information about surface morphology. On the other hand, electrical characterizations like Current Voltage (IV) measurement in dark and light and test with laser are performed to have a better understanding of the working of the detector prototype. All these preliminary tests of the detector will be presented.

Keywords: particle detector, rutile titanium dioxide, thermal evaporation, wide bandgap semiconductors

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1111 Targeting Tumour Survival and Angiogenic Migration after Radiosensitization with an Estrone Analogue in an in vitro Bone Metastasis Model

Authors: Jolene M. Helena, Annie M. Joubert, Peace Mabeta, Magdalena Coetzee, Roy Lakier, Anne E. Mercier

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Targeting the distant tumour and its microenvironment whilst preserving bone density is important in improving the outcomes of patients with bone metastases. 2-Ethyl-3-O-sulphamoyl-estra1,3,5(10)16-tetraene (ESE-16) is an in-silico-designed 2- methoxyestradiol analogue which aimed at enhancing the parent compound’s cytotoxicity and providing a more favourable pharmacokinetic profile. In this study, the potential radiosensitization effects of ESE-16 were investigated in an in vitro bone metastasis model consisting of murine pre-osteoblastic (MC3T3-E1) and pre-osteoclastic (RAW 264.7) bone cells, metastatic prostate (DU 145) and breast (MDA-MB-231) cancer cells, as well as human umbilical vein endothelial cells (HUVECs). Cytotoxicity studies were conducted on all cell lines via spectrophotometric quantification of 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide. The experimental set-up consisted of flow cytometric analysis of cell cycle progression and apoptosis detection (Annexin V-fluorescein isothiocyanate) to determine the lowest ESE-16 and radiation doses to induce apoptosis and significantly reduce cell viability. Subsequent experiments entailed a 24-hour low-dose ESE-16-exposure followed by a single dose of radiation. Termination proceeded 2, 24 or 48 hours thereafter. The effect of the combination treatment was investigated on osteoclasts via tartrate-resistant acid phosphatase (TRAP) activity- and actin ring formation assays. Tumour cell experiments included investigation of mitotic indices via haematoxylin and eosin staining; pro-apoptotic signalling via spectrophotometric quantification of caspase 3; deoxyribonucleic acid (DNA) damage via micronuclei analysis and histone H2A.X phosphorylation (γ-H2A.X); and Western blot analyses of bone morphogenetic protein-7 and matrix metalloproteinase-9. HUVEC experiments included flow cytometric quantification of cell cycle progression and free radical production; fluorescent examination of cytoskeletal morphology; invasion and migration studies on an xCELLigence platform; and Western blot analyses of hypoxia-inducible factor 1-alpha and vascular endothelial growth factor receptor 1 and 2. Tumour cells yielded half-maximal growth inhibitory concentration (GI50) values in the nanomolar range. ESE-16 concentrations of 235 nM (DU 145) and 176 nM (MDA-MB-231) and a radiation dose of 4 Gy were found to be significant in cell cycle and apoptosis experiments. Bone and endothelial cells were exposed to the same doses as DU 145 cells. Cytotoxicity studies on bone cells reported that RAW 264.7 cells were more sensitive to the combination treatment than MC3T3-E1 cells. Mature osteoclasts were more sensitive than pre-osteoclasts with respect to TRAP activity. However, actin ring morphology was retained. The mitotic arrest was evident in tumour and endothelial cells in the mitotic index and cell cycle experiments. Increased caspase 3 activity and superoxide production indicated pro-apoptotic signalling in tumour and endothelial cells. Increased micronuclei numbers and γ-H2A.X foci indicated increased DNA damage in tumour cells. Compromised actin and tubulin morphologies and decreased invasion and migration were observed in endothelial cells. Western blot analyses revealed reduced metastatic and angiogenic signalling. ESE-16-induced radiosensitization inhibits metastatic signalling and tumour cell survival whilst preferentially preserving bone cells. This low-dose combination treatment strategy may promote the quality of life of patients with metastatic bone disease. Future studies will include 3-dimensional in-vitro and murine in-vivo models.

Keywords: angiogenesis, apoptosis, bone metastasis, cancer, cell migration, cytoskeleton, DNA damage, ESE-16, radiosensitization.

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1110 A Comprehensive Survey on Machine Learning Techniques and User Authentication Approaches for Credit Card Fraud Detection

Authors: Niloofar Yousefi, Marie Alaghband, Ivan Garibay

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With the increase of credit card usage, the volume of credit card misuse also has significantly increased, which may cause appreciable financial losses for both credit card holders and financial organizations issuing credit cards. As a result, financial organizations are working hard on developing and deploying credit card fraud detection methods, in order to adapt to ever-evolving, increasingly sophisticated defrauding strategies and identifying illicit transactions as quickly as possible to protect themselves and their customers. Compounding on the complex nature of such adverse strategies, credit card fraudulent activities are rare events compared to the number of legitimate transactions. Hence, the challenge to develop fraud detection that are accurate and efficient is substantially intensified and, as a consequence, credit card fraud detection has lately become a very active area of research. In this work, we provide a survey of current techniques most relevant to the problem of credit card fraud detection. We carry out our survey in two main parts. In the first part, we focus on studies utilizing classical machine learning models, which mostly employ traditional transnational features to make fraud predictions. These models typically rely on some static physical characteristics, such as what the user knows (knowledge-based method), or what he/she has access to (object-based method). In the second part of our survey, we review more advanced techniques of user authentication, which use behavioral biometrics to identify an individual based on his/her unique behavior while he/she is interacting with his/her electronic devices. These approaches rely on how people behave (instead of what they do), which cannot be easily forged. By providing an overview of current approaches and the results reported in the literature, this survey aims to drive the future research agenda for the community in order to develop more accurate, reliable and scalable models of credit card fraud detection.

Keywords: Credit Card Fraud Detection, User Authentication, Behavioral Biometrics, Machine Learning, Literature Survey

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1109 Ferulic Acid-Grafted Chitosan: Thermal Stability and Feasibility as an Antioxidant for Active Biodegradable Packaging Film

Authors: Sarekha Woranuch, Rangrong Yoksan

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Active packaging has been developed based on the incorporation of certain additives, in particular antimicrobial and antioxidant agents, into packaging systems to maintain or extend product quality and shelf-life. Ferulic acid is one of the most effective natural phenolic antioxidants, which has been used in food, pharmaceutical and active packaging film applications. However, most phenolic compounds are sensitive to oxygen, light and heat; its activities are thus lost during product formulation and processing. Grafting ferulic acid onto polymer is an alternative to reduce its loss under thermal processes. Therefore, the objectives of the present research were to study the thermal stability of ferulic acid after grafting onto chitosan, and to investigate the possibility of using ferulic acid-grafted chitosan (FA-g-CTS) as an antioxidant for active biodegradable packaging film. FA-g-CTS was incorporated into biodegradable film via a two-step process, i.e. compounding extrusion at temperature up to 150 °C followed by blown film extrusion at temperature up to 175 °C. Although incorporating FA-g-CTS with a content of 0.02–0.16% (w/w) caused decreased water vapor barrier property and reduced extensibility, the films showed improved oxygen barrier property and antioxidant activity. Radical scavenging activity and reducing power of the film containing FA-g-CTS with a content of 0.04% (w/w) were higher than that of the naked film about 254% and 94%, respectively. Tensile strength and rigidity of the films were not significantly affected by adding FA-g-CTS with a content of 0.02–0.08% (w/w). The results indicated that FA-g-CTS could be potentially used as an antioxidant for active packaging film.

Keywords: active packaging film, antioxidant activity, chitosan, ferulic acid

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1108 Assessment of Students Skills in Error Detection in SQL Classes using Rubric Framework - An Empirical Study

Authors: Dirson Santos De Campos, Deller James Ferreira, Anderson Cavalcante Gonçalves, Uyara Ferreira Silva

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Rubrics to learning research provide many evaluation criteria and expected performance standards linked to defined student activity for learning and pedagogical objectives. Despite the rubric being used in education at all levels, academic literature on rubrics as a tool to support research in SQL Education is quite rare. There is a large class of SQL queries is syntactically correct, but certainly, not all are semantically correct. Detecting and correcting errors is a recurring problem in SQL education. In this paper, we usthe Rubric Abstract Framework (RAF), which consists of steps, that allows us to map the information to measure student performance guided by didactic objectives defined by the teacher as long as it is contextualized domain modeling by rubric. An empirical study was done that demonstrates how rubrics can mitigate student difficulties in finding logical errors and easing teacher workload in SQL education. Detecting and correcting logical errors is an important skill for students. Researchers have proposed several ways to improve SQL education because understanding this paradigm skills are crucial in software engineering and computer science. The RAF instantiation was using in an empirical study developed during the COVID-19 pandemic in database course. The pandemic transformed face-to-face and remote education, without presential classes. The lab activities were conducted remotely, which hinders the teaching-learning process, in particular for this research, in verifying the evidence or statements of knowledge, skills, and abilities (KSAs) of students. Various research in academia and industry involved databases. The innovation proposed in this paper is the approach used where the results obtained when using rubrics to map logical errors in query formulation have been analyzed with gains obtained by students empirically verified. The research approach can be used in the post-pandemic period in both classroom and distance learning.

Keywords: rubric, logical error, structured query language (SQL), empirical study, SQL education

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1107 Effects of Magnetization Patterns on Characteristics of Permanent Magnet Linear Synchronous Generator for Wave Energy Converter Applications

Authors: Sung-Won Seo, Jang-Young Choi

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The rare earth magnets used in synchronous generators offer many advantages, including high efficiency, greatly reduced the size, and weight. The permanent magnet linear synchronous generator (PMLSG) allows for direct drive without the need for a mechanical device. Therefore, the PMLSG is well suited to translational applications, such as wave energy converters and free piston energy converters. This manuscript compares the effects of different magnetization patterns on the characteristics of double-sided PMLSGs in slotless stator structures. The Halbach array has a higher flux density in air-gap than the Vertical array, and the advantages of its performance and efficiency are widely known. To verify the advantage of Halbach array, we apply a finite element method (FEM) and analytical method. In general, a FEM and an analytical method are used in the electromagnetic analysis for determining model characteristics, and the FEM is preferable to magnetic field analysis. However, the FEM is often slow and inflexible. On the other hand, the analytical method requires little time and produces accurate analysis of the magnetic field. Therefore, the flux density in air-gap and the Back-EMF can be obtained by FEM. In addition, the results from the analytical method correspond well with the FEM results. The model of the Halbach array reveals less copper loss than the model of the Vertical array, because of the Halbach array’s high output power density. The model of the Vertical array is lower core loss than the model of Halbach array, because of the lower flux density in air-gap. Therefore, the current density in the Vertical model is higher for identical power output. The completed manuscript will include the magnetic field characteristics and structural features of both models, comparing various results, and specific comparative analysis will be presented for the determination of the best model for application in a wave energy converting system.

Keywords: wave energy converter, permanent magnet linear synchronous generator, finite element method, analytical method

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1106 Current Approach in Biodosimetry: Electrochemical Detection of DNA Damage

Authors: Marcela Jelicova, Anna Lierova, Zuzana Sinkorova, Radovan Metelka

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At present, electrochemical methods are used in various research fields, especially for analysis of biological molecules. The fact offers the possibility of using the detection of oxidative damage induced indirectly by γ rays in DNA in biodosimentry. The main goal of our study is to optimize the detection of 8-hydroxyguanine by differential pulse voltammetry. The level of this stable and specific indicator of DNA damage could be determined in DNA isolated from peripheral blood lymphocytes, plasma or urine of irradiated individuals. Screen-printed carbon electrodes modified with carboxy-functionalized multi-walled carbon nanotubes were utilized for highly sensitive electrochemical detection of 8-hydroxyguanine. Electrochemical oxidation of 8-hydroxoguanine monitored by differential pulse voltammetry was found pH-dependent and the most intensive signal was recorded at pH 7. After recalculating the current density, several times higher sensitivity was attained in comparison with already published results, which were obtained using screen-printed carbon electrodes with unmodified carbon ink. Subsequently, the modified electrochemical technique was used for the detection of 8-hydroxoguanine in calf thymus DNA samples irradiated by 60Co gamma source in the dose range from 0.5 to 20 Gy using by various types of sample pretreatment and measurement conditions. This method could serve for fast retrospective quantification of absorbed dose in cases of accidental exposure to ionizing radiation and may play an important role in biodosimetry.

Keywords: biodosimetry, electrochemical detection, voltametry, 8-hydroxyguanine

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1105 Segmental Dynamics of Poly(Alkyl Methacrylate) Chain in Ultra-Thin Spin-Cast Films

Authors: Hiroyuki Aoki

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Polymeric materials are often used in a form of thin film such as food wrap and surface coating. In such the applications, polymer films thinner than 100 nm have been often used. The thickness of such the ultra-thin film is less than the unperturbed size of a polymer chain; therefore, the polymer chain in an ultra-thin film is strongly constrained. However, the details on the constrained dynamics of polymer molecules in ultra-thin films are still unclear. In the current study, the segmental dynamics of single polymer chain was directly investigated by fluorescence microscopy. The individual chains of poly(alkyl methacrylate) labeled by a perylenediimide dye molecule were observed by a highly sensitive fluorescence microscope in a defocus condition. The translational and rotational diffusion of the center segment in a single polymer chain was directly analyzed. The segmental motion in a thin film with a thickness of 10 nm was found to be suppressed compared to that in a bulk state. The detailed analysis of the molecular motion revealed that the diffusion rate of the in-plane rotation was similar to the thin film and the bulk; on the other hand, the out-of-plane motion was restricted in a thin film. This result indicates that the spatial restriction in an ultra-thin film thinner than the unperturbed chain dimension alters the dynamics of individual molecules in a polymer system.

Keywords: polymer materials, single molecule, molecular motion, fluorescence microscopy, super-resolution techniques

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1104 Investigation on the Effect of Titanium (Ti) Plus Boron (B) Addition to the Mg-AZ31 Alloy in the as Cast and After Extrusion on Its Metallurgical and Mechanical Characteristics

Authors: Adnan I. O. Zaid, Raghad S. Hemeimat

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Magnesium - aluminum alloys are versatile materials which are used in manufacturing a number of engineering and industrial parts in the automobile and aircraft industries due to their strength – to –weight -ratio. Against these preferable characteristics, magnesium is difficult to deform at room temperature therefore it is alloyed with other elements mainly Aluminum and Zinc to add some required properties particularly for their high strength - to -weight ratio. Mg and its alloys oxidize rapidly therefore care should be taken during melting or machining them; but they are not fire hazardous. Grain refinement is an important technology to improve the mechanical properties and the micro structure uniformity of the alloys. Grain refinement has been introduced in early fifties; when Cibula showed that the presence of Ti, and Ti+ B, produced a great refining effect in Al. since then it became an industrial practice to grain refine Al. Most of the published work on grain refinement was directed toward grain refining Al and Zinc alloys; however, the effect of the addition of rare earth material on the grain size or the mechanical behavior of Mg alloys has not been previously investigated. This forms the main objective of the research work; where, the effect of Ti addition on the grain size, mechanical behavior, ductility, and the extrusion force & energy consumed in forward extrusion of Mg-AZ31 alloy is investigated and discussed in two conditions, first in the as cast condition and the second after extrusion. It was found that addition of Ti to Mg- AZ31 alloy has resulted in reduction of its grain size by 14%; the reduction in grain size after extrusion was much higher. However the increase in Vicker’s hardness was 3% after the addition of Ti in the as cast condition, and higher values for Vicker’s hardness were achieved after extrusion. Furthermore, an increase in the strength coefficient by 36% was achieved with the addition of Ti to Mg-AZ31 alloy in the as cast condition. Similarly, the work hardening index was also increased indicating an enhancement of the ductility and formability. As for the extrusion process, it was found that the force and energy required for the extrusion were both reduced by 57% and 59% with the addition of Ti.

Keywords: cast condition, direct extrusion, ductility, MgAZ31 alloy, super - plasticity

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1103 Evaluation of the Microscopic-Observation Drug-Susceptibility Assay Drugs Concentration for Detection of Multidrug-Resistant Tuberculosis

Authors: Anita, Sari Septiani Tangke, Rusdina Bte Ladju, Nasrum Massi

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New diagnostic tools are urgently needed to interrupt the transmission of tuberculosis and multidrug-resistant tuberculosis. The microscopic-observation drug-susceptibility (MODS) assay is a rapid, accurate and simple liquid culture method to detect multidrug-resistant tuberculosis (MDR-TB). MODS were evaluated to determine a lower and same concentration of isoniazid and rifampin for detection of MDR-TB. Direct drug-susceptibility testing was performed with the use of the MODS assay. Drug-sensitive control strains were tested daily. The drug concentrations that used for both isoniazid and rifampin were at the same concentration: 0.16, 0.08 and 0.04μg per milliliter. We tested 56 M. tuberculosis clinical isolates and the control strains M. tuberculosis H37RV. All concentration showed same result. Of 53 M. tuberculosis clinical isolates, 14 were MDR-TB, 38 were susceptible with isoniazid and rifampin, 1 was resistant with isoniazid only. Drug-susceptibility testing was performed with the use of the proportion method using Mycobacteria Growth Indicator Tube (MGIT) system as reference. The result of MODS assay using lower concentration was significance (P<0.001) compare with the reference methods. A lower and same concentration of isoniazid and rifampin can be used to detect MDR-TB. Operational cost and application can be more efficient and easier in resource-limited environments. However, additional studies evaluating the MODS using lower and same concentration of isoniazid and rifampin must be conducted with a larger number of clinical isolates.

Keywords: isoniazid, MODS assay, MDR-TB, rifampin

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1102 Patient Outcomes Following Out-of-Hospital Cardiac Arrest

Authors: Scott Ashby, Emily Granger, Mark Connellan

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Background: In-hospital management of Out-of-Hospital Cardiac Arrest (OHCA) is complex as the aetiologies are varied. Acute coronary angiography has been shown to improve outcomes for patients with coronary occlusion as the cause; however, these patients are difficult to identify. ECG results may help identify these patients, but the accuracy of this diagnostic test is under debate, and requires further investigation. Methods: Arrest and hospital management information was collated retrospectively for OHCA patients who presented to a single clinical site between 2009 and 2013. Angiography results were then collected and checked for significance with survival to discharge. The presence of a severe lesion (>70%) was then compared to categorised ECG findings, and the accuracy of the test was calculated. Results: 104 patients were included in this study, 44 survived to discharge, 52 died and 8 were transferred to other clinical sites. Angiography appears to significantly correlate with survival to discharge. ECG showed 54.8% sensitivity for detecting the presence of a severe lesion within the group that received angiography. A combined criterion including any ECG pathology showed 100% sensitivity and negative predictive value, however, a low specificity and positive predictive value. Conclusion: In the cohort investigated, ST elevation on ECG is not a sensitive enough screening test to be used to determine whether OHCA patients have coronary stenosis as the likely cause of their arrest, and more investigation into whether screening with a combined ECG criterion, or whether all patients should receive angiography routinely following OHCA is needed.

Keywords: out of hospital cardiac arrest, coronary angiography, resuscitation, emergency medicine

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1101 Risk Factors for High Resistance of Ciprofloxacin Against Escherichia coli in Complicated Urinary Tract Infection

Authors: Liaqat Ali, Khalid Farooq, Shafieullah Khan, Nasir Orakzai, Qudratullah

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Objectives: To determine the risk factors for high resistance of ciprofloxacin in complicated urinary tract infections. Materials and Methods: It is an analytical study that was conducted in the department of Urology (Team ‘C’) at Institute of Kidney Diseases Hayatabad Peshawar from 1st June 2012 till 31st December 2012. Total numbers of 100 patients with complicated UTI was selected in the study. Multivariate analysis and linear regression were performed for the detection of risk factors. All the data was recorded on structured Proforma and was analyzed on SPSS version 17. Results: The mean age of the patient was 55.6 years (Range 3-82 years). 62 patients were male while 38 patients were female. 66 isolates of E-Coli were found sensitive to ciprofloxacin while 34 isolates were found Resistant for ciprofloxacin. Using multivariate analysis and linear regression, an increasing age above 50 (p=0.002) History of urinary catheterization especially for bladder outflow obstruction (p=0.001) and previous multiple use of ciprofloxacin (p=0.001) and poor brand of ciprofloxacin were found to be independent risk factors for high resistance of ciprofloxacin. Conclusion: UTI is common illness across the globe with increasing trend of antimicrobial resistance for ciprofloxacin against E Coli in complicated UTI. The risk factors for emerging resistance are increasing age, urinary catheterization and multiple use and poor brand of ciprofloxacin.

Keywords: urinary tract infection, ciprofloxacin, urethral catheterization, antimicrobial resistance

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1100 Comet Assay: A Promising Tool for the Risk Assessment and Clinical Management of Head and Neck Tumors

Authors: Sarim Ahmad

Abstract:

The Single Cell Gel Electrophoresis Assay (SCGE, known as comet assay) is a potential, uncomplicated, sensitive and state-of-the-art technique for quantitating DNA damage at individual cell level and repair from in vivo and in vitro samples of eukaryotic cells and some prokaryotic cells, being popular in its widespread use in various areas including human biomonitoring, genotoxicology, ecological monitoring and as a tool for research into DNA damage or repair in different cell types in response to a range of DNA damaging agents, cancer risk and therapy. The method involves the encapsulation of cells in a low-melting-point agarose suspension, lysis of the cells in neutral or alkaline (pH > 13) conditions, and electrophoresis of the suspended lysed cells, resulting in structures resembling comets as observed by fluorescence microscopy; the intensity of the comet tail relative to the head reflects the number of DNA breaks. The likely basis for this is that loops containing a break lose their supercoiling and become free to extend towards the anode. This is followed by visual analysis with staining of DNA and calculating fluorescence to determine the extent of DNA damage. This can be performed by manual scoring or automatically by imaging software. The assay can, therefore, predict an individual’s tumor sensitivity to radiation and various chemotherapeutic drugs and further assess the oxidative stress within tumors and to detect the extent of DNA damage in various cancerous and precancerous lesions of oral cavity.

Keywords: comet assay, single cell gel electrophoresis, DNA damage, early detection test

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1099 Determination of Acid Volatile Sulfides–Simultaneously Extracted Metal Relationship and Toxicity in Contaminated Sediment Layer in Mid-Black Sea Coasts

Authors: Arife Simsek, Gulfem Bakan

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Sediment refers to the accumulation of varying amounts of sediment material in natural waters and the formation of bottom sludge. Sediments are the most important sources of pollutants as well as important future sources and carriers of pollutants. The accumulation of pollutants in sediments can cause serious environmental problems for the surrounding areas. Heavy metals (such as Cr, Cd, Al, Pb, Cu, Al, Zn) disrupt the water quality, affect the useful use of sediment, affect the ecosystem and have a toxic effect on the life of the sediment layer. This effect, which accumulates in the aquatic organisms, can enter the human body with the food chain and affect health seriously. Potential metal toxicity can be determined by comparing acid volatile sulfides (AVS) – simultaneously extracted metal (SEM) ratio in anoxic sediments to determine the effect of metals. Determination of the concentration of SEM and AVS is useful in screening sediments for potential toxicity due to the high metal concentration. In the case of SEM/AVS < 0 (anoxic sediment); in terms of AVS biomass production, its toxicity can be controlled. No toxic effects may be observed when SEM / AVS < 0. SEM / AVS > 0 (in the case of oxic sediment); metals with sensitive fraction such as Cu, As, Ag, Zn are stored. In this study, AVS and SEM measurements of sediment samples collected from five different points in the district of Tekkeköy in Samsun province were performed. The SEM - AVS ratio was greater than 0 in all samples. Therefore, it is necessary to test the toxicity against the risks that may occur in the ecosystem.

Keywords: AVS-SEM, Black Sea, heavy metal, sediment, toxicity

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1098 Reviewing the Effect of Healing Design on Mental Health Establishments in the Context of India

Authors: Aratrika Sarkar, Jayita Guha Niyogi

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This paper focuses on the application of general healing design theories to modulate them into case-specific and contextual design considerations. Existing literature focuses on the relationship between architecture and mental health. Primary case studies are selected in India to focus on the effect of a specific location on design considerations. They are qualitatively analysed to further contextualise the inferences from the literature study. An academic project is cited as an example to apply the learnings from the study and understand the influence of various parameters on the design process for further conclusion. Literature studies, case studies and hypothetical design applications helped in finding the different ways of achieving the similar goal of a sensitive approach toward mental health. Along with salutogenic parameters, category of establishment, age group, location of the site and user preference plays a crucial role in the design process. Design of mental health establishments, especially in India, has to involve transparency between stakeholders and users. Owing to different climatic zones and diverse sociocultural traditions, the approach toward healing should adapt accordingly. It should be an effort towards striking a balance between contradictory elements of healing design and resolving the dilemmas with sensitivity and consensus. Lastly, the design should not force a person towards communication or companionship but rather let the person realise that naturally through the healing process.

Keywords: contextual healing design, deinstitutionalisation, Indian mental healthcare establishments, environmental psychology, salutogenesis, therapeutic design

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1097 River Offtake Management Using Mathematical Modelling Tool: A Case Study of the Gorai River, Bangladesh

Authors: Sarwat Jahan, Asker Rajin Rahman

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Management of offtake of any fluvial river is very sensitive in terms of long-term sustainability where the variation of water flow and sediment transport range are wide enough throughout a hydrological year. The Gorai River is a major distributary of the Ganges River in Bangladesh and is termed as a primary source of fresh water for the South-West part of the country. Every year, significant siltation of the Gorai offtake disconnects it from the Ganges during the dry season. As a result, the socio-economic and environmental condition of the downstream areas has been deteriorating for a few decades. To improve the overall situation of the Gorai offtake and its dependent areas, a study has been conducted by the Institute of Water Modelling, Bangladesh, in 2022. Using the mathematical morphological modeling tool MIKE 21C of DHI Water & Environment, Denmark, simulated results revealed the need for dredging/river training structures for offtake management at the Gorai offtake to ensure significant dry season flow towards the downstream. The dry season flow is found to increase significantly with the proposed river interventions, which also improves the environmental conditions in terms of salinity of the South-West zone of the country. This paper summarizes the primary findings of the analyzed results of the developed mathematical model for improving the existing condition of the Gorai River.

Keywords: Gorai river, mathematical modelling, offtake, siltation, salinity

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1096 Management of First Trimester Miscarriage

Authors: Madeleine Cox

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Objective; analyse patient choices in management of first trimester miscarriage, rates of complications including repeat procedure. Design: all first trimester miscarriages from a tertiary institution on the Gold Coast in a 6 month time frame (July to December 2021) were reviewed, including choice of management, histopathology, any representations or admissions, and potential complications. Results: a total of 224 first trimester miscarriages were identified. Of these, 183 (81%) opted to have surgical management in the first instance. Of the remaining patients, 18 (8%) opted to have medical management, and 28 (12.5%) opted to have expectant management. In total, 33(15%) patients required a repeat treatment for retained products. 1 had medical management for a small volume PROC post suction curette. A significant number of these patients initially opted for medical management but then elected to have shorter follow up than usual and went on to have retained products noted. 5 women who had small volumes of RPOC post medical or surgical management had repeat suction curette, however, had very small volumes of products on scan and on curette and may have had a good result with repeated misoprostol administration. It is important to note that whilst a common procedure, suction curettes are not without risk. 2 women had significant blood loss of 1L and 1.5L. A third women had a uterine perforation, a rare but recognised complication, she went on to require a laparoscopy which identified a small serosal bowel injury which was closed by the colorectal team. Conclusion: Management of first trimester miscarriage should be guided by patient preference. It is important to be able to provide patients with their choice of management, however, it is also important to have a good understanding of the risks of each management choice, chances of repeated procedure, appropriate time frame for follow up. Women who choose to undertake medical or expectant management should be supported through this time, with appropriate time frame between taking misoprostol and repeat scan so that the true effects can be evaluated. Patients returning for scans within 2-3 days are more likely to be booked for further surgery, however, may reflect patients who did not have adequate counselling or simply changed their mind on their preferred management options.

Keywords: miscarriage, gynaecology, obstetrics, first trimester

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1095 Hardware Implementation on Field Programmable Gate Array of Two-Stage Algorithm for Rough Set Reduct Generation

Authors: Tomasz Grzes, Maciej Kopczynski, Jaroslaw Stepaniuk

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The rough sets theory developed by Prof. Z. Pawlak is one of the tools that can be used in the intelligent systems for data analysis and processing. Banking, medicine, image recognition and security are among the possible fields of utilization. In all these fields, the amount of the collected data is increasing quickly, but with the increase of the data, the computation speed becomes the critical factor. Data reduction is one of the solutions to this problem. Removing the redundancy in the rough sets can be achieved with the reduct. A lot of algorithms of generating the reduct were developed, but most of them are only software implementations, therefore have many limitations. Microprocessor uses the fixed word length, consumes a lot of time for either fetching as well as processing of the instruction and data; consequently, the software based implementations are relatively slow. Hardware systems don’t have these limitations and can process the data faster than a software. Reduct is the subset of the decision attributes that provides the discernibility of the objects. For the given decision table there can be more than one reduct. Core is the set of all indispensable condition attributes. None of its elements can be removed without affecting the classification power of all condition attributes. Moreover, every reduct consists of all the attributes from the core. In this paper, the hardware implementation of the two-stage greedy algorithm to find the one reduct is presented. The decision table is used as an input. Output of the algorithm is the superreduct which is the reduct with some additional removable attributes. First stage of the algorithm is calculating the core using the discernibility matrix. Second stage is generating the superreduct by enriching the core with the most common attributes, i.e., attributes that are more frequent in the decision table. Described above algorithm has two disadvantages: i) generating the superreduct instead of reduct, ii) additional first stage may be unnecessary if the core is empty. But for the systems focused on the fast computation of the reduct the first disadvantage is not the key problem. The core calculation can be achieved with a combinational logic block, and thus add respectively little time to the whole process. Algorithm presented in this paper was implemented in Field Programmable Gate Array (FPGA) as a digital device consisting of blocks that process the data in a single step. Calculating the core is done by the comparators connected to the block called 'singleton detector', which detects if the input word contains only single 'one'. Calculating the number of occurrences of the attribute is performed in the combinational block made up of the cascade of the adders. The superreduct generation process is iterative and thus needs the sequential circuit for controlling the calculations. For the research purpose, the algorithm was also implemented in C language and run on a PC. The times of execution of the reduct calculation in a hardware and software were considered. Results show increase in the speed of data processing.

Keywords: data reduction, digital systems design, field programmable gate array (FPGA), reduct, rough set

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1094 In vitro Modeling of Aniridia-Related Keratopathy by the Use of Crispr/Cas9 on Limbal Epithelial Cells and Rescue

Authors: Daniel Aberdam

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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.

Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy

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1093 Laparoscopic Uterovaginal Anastomosis in Cervicovaginal Agenesis

Authors: Anamika Choudhary, Neha Qurrat Ain

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Background: Congenital agenesis of uterine cervix is a rare anomaly often associated with partial or complete agenesis of vagina. Here is a case report of a 14 year old girl who presented with primary amenorrhea and cyclical abdominal pain since last one year with suprapubic mass palpable. On examination complete absence of a vagina was found, and ultrasound along with magnetic resonance imaging (MRI) suggested cervicovaginal agenesis associated with cryptomenorrhea, which resulted in hematometra and b/l hematosalpinx with pelvic endometriosis. After proper counseling regarding anastomosis failure and the need for future laprotomy or hysterectomy, the patient planned for laparoscopic uterovaginal anastomosis with modified McIndoe vaginoplasty with split skin graft. Case Summary: Chief complaint: The 14 year old girl presented with primary amenorrhea and cyclical abdominal pain. Diagnosis:On history, examination and investigations we made differential diagnoses of cervicovaginal agenesis, cervicovaginal atresia. Post operatively, we concluded it’s a cervicovaginal agenesis. Intervention: Laparoscopic uterovaginal anastomosis was done, and neovagina was created using split skin graft from the thigh and silicone stent. The graft was kept patent, and restenosis was prevented using a dental mould as vaginal dilator. Outcome: Postoperatively 1 year follow-up has been done. We have observed successful uterovaginal anastomosis and good uptake of graft. We also observed the resumption of normal menstrual bleeding. Currently, there has been no restenosis, abnormal vaginal discharge and decreased dysmenorrhea. Conclusion: Laparoscopic-assisted uterovaginal anastomosis can be the treatment of choice in patients with cervical agenesis and atresia instead of hysterectomy, thereby preserving the reproductive function. This conservative approach has better outcomes, as stated in the procedure below. The procedure is successful insofar as the resumption of menstrual function. However, long-term reproductive outcomes, progression of endometriosis, functioning of fallopian tubes, and sexual life in these girls will require further follow-up.

Keywords: cervicovaginal agenesis, uterovaginal anastomosis, dental mould, silicon stent

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