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Статті в журналах з теми "MLPANN"
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Mirbagheri, Sayed Ahmad, Majid Bagheri, Majid Ehteshami, Zahra Bagheri, and Masoud Pourasghar. "MODELING OF MIXED LIQUOR VOLATILE SUSPENDED SOLIDS AND PERFORMANCE EVALUATION FOR A SEQUENCING BATCH REACTOR." Journal of Urban and Environmental Engineering 9, no. 1 (December 27, 2015): 54–65. http://dx.doi.org/10.4090/juee.2015.v9n1.054065.
Повний текст джерелаАнотація:
This study examined carbon, nitrogen and phosphorous removal from municipal wastewater in a sequencing batch reactor and biokinetic coefficients were evaluated according to results of BOD and COD. Furthermore, the MLVSS in the aeration reactor was modeled by using multilayer perceptron and radial basis function artificial neural networks (MLPANN and RBFANN). The experiments were performed so that the cell retention time, filling time and intensity of aeration were (5, 10 and 15 d), (1, 2 and 3 h) and (weak, medium and strong) respectively. The result indicated that with cell retention time of 15 d, filling time of 1 h, aeration time of 6 h and settling time of 3 h the HRT is optimized at 10 h. The BOD5, COD, TP, TN and removal efficiencies were 97.13%, 94.58%, 94.27%, 89.7% and 92.75% respectively. The yield coefficient (Y), decay coefficient (Kd), maximum specific growth rate (K) and saturation constant (Ks) were 6.22 mgVSS/mgCOD, 0.002 1/d, 0.029 1/d and 20 mg COD/L according to COD experimental data. The values of the biokinetic coefficients were found to be as follows: Y = 10.45 mgVSS/mgBOD, Kd = 0.01 1/d, 0.014 1/d and 3.38 mgBOD/L according to BOD5 experimental data. The training procedures for simulation of MLVSS were highly collaborated for both RBFANN and MLPANN. The train and test models for both MLPANN and RBFANN demonstrated perfectly matched results between the experimental and the simulated values of MLVSS. The values of RMSE for train and test (verification) models obtained by MLPANN were 31.82 and 40.25 mg/L respectively, and the value of R2 was 0.99 for both models. The values of RMSE for train and test models obtained by MLPANN were 69.04 and 43.87 mg/L respectively, and the value of R2 was 0.99 for both models. It was observed that the MLPANN has stronger approximation and generalization ability than the RBFANN with regard to our experimental data for MLVSS.
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Mirbagheri, Sayed Ahmad, Majid Bagheri, Majid Ehteshami, Zahra Bagheri, and Masoud Pourasghar. "MODELING OF MIXED LIQUOR VOLATILE SUSPENDED SOLIDS AND PERFORMANCE EVALUATION FOR A SEQUENCING BATCH REACTOR." Journal of Urban and Environmental Engineering 9, no. 1 (December 27, 2015): 54–65. http://dx.doi.org/10.4090/juee.2015.v9n1.54-65.
Повний текст джерелаАнотація:
This study examined carbon, nitrogen and phosphorous removal from municipal wastewater in a sequencing batch reactor and biokinetic coefficients were evaluated according to results of BOD and COD. Furthermore, the MLVSS in the aeration reactor was modeled by using multilayer perceptron and radial basis function artificial neural networks (MLPANN and RBFANN). The experiments were performed so that the cell retention time, filling time and intensity of aeration were (5, 10 and 15 d), (1, 2 and 3 h) and (weak, medium and strong) respectively. The result indicated that with cell retention time of 15 d, filling time of 1 h, aeration time of 6 h and settling time of 3 h the HRT is optimized at 10 h. The BOD5, COD, TP, TN and removal efficiencies were 97.13%, 94.58%, 94.27%, 89.7% and 92.75% respectively. The yield coefficient (Y), decay coefficient (Kd), maximum specific growth rate (K) and saturation constant (Ks) were 6.22 mgVSS/mgCOD, 0.002 1/d, 0.029 1/d and 20 mg COD/L according to COD experimental data. The values of the biokinetic coefficients were found to be as follows: Y = 10.45 mgVSS/mgBOD, Kd = 0.01 1/d, 0.014 1/d and 3.38 mgBOD/L according to BOD5 experimental data. The training procedures for simulation of MLVSS were highly collaborated for both RBFANN and MLPANN. The train and test models for both MLPANN and RBFANN demonstrated perfectly matched results between the experimental and the simulated values of MLVSS. The values of RMSE for train and test (verification) models obtained by MLPANN were 31.82 and 40.25 mg/L respectively, and the value of R2 was 0.99 for both models. The values of RMSE for train and test models obtained by MLPANN were 69.04 and 43.87 mg/L respectively, and the value of R2 was 0.99 for both models. It was observed that the MLPANN has stronger approximation and generalization ability than the RBFANN with regard to our experimental data for MLVSS.
3
Alizamir, Meysam, Zahra Kazemi, Zohre Kazemi, Majid Kermani, Sungwon Kim, Salim Heddam, Ozgur Kisi, and Il-Moon Chung. "Investigating Landfill Leachate and Groundwater Quality Prediction Using a Robust Integrated Artificial Intelligence Model: Grey Wolf Metaheuristic Optimization Algorithm and Extreme Learning Machine." Water 15, no. 13 (July 4, 2023): 2453. http://dx.doi.org/10.3390/w15132453.
Повний текст джерелаАнотація:
The likelihood of surface water and groundwater contamination is higher in regions close to landfills due to the possibility of leachate percolation, which is a potential source of pollution. Therefore, proposing a reliable framework for monitoring leachate and groundwater parameters is an essential task for the managers and authorities of water quality control. For this purpose, an efficient hybrid artificial intelligence model based on grey wolf metaheuristic optimization algorithm and extreme learning machine (ELM-GWO) is used for predicting landfill leachate quality (COD and BOD5) and groundwater quality (turbidity and EC) at the Saravan landfill, Rasht, Iran. In this study, leachate and groundwater samples were collected from the Saravan landfill and monitoring wells. Moreover, the concentration of different physico-chemical parameters and heavy metal concentration in leachate (Cd, Cr, Cu, Fe, Ni, Pb, Mn, Zn, turbidity, Ca, Na, NO3, Cl, K, COD, and BOD5) and in groundwater (Cd, Cr, Cu, Fe, Ni, Pb, Mn, Zn, turbidity, EC, TDS, pH, Cl, Na, NO3, and K). The results obtained from ELM-GWO were compared with four different artificial intelligence models: multivariate adaptive regression splines (MARS), extreme learning machine (ELM), multilayer perceptron artificial neural network (MLPANN), and multilayer perceptron artificial neural network integrated with grey wolf metaheuristic optimization algorithm (MLPANN-GWO). The results of this study confirm that ELM-GWO considerably enhanced the predictive performance of the MLPANN-GWO, ELM, MLPANN, and MARS models in terms of the root-mean-square error, respectively, by 43.07%, 73.88%, 74.5%, and 88.55% for COD; 23.91%, 59.31%, 62.85%, and 77.71% for BOD5; 14.08%, 47.86%, 53.43%, and 57.04% for turbidity; and 38.57%, 59.64%, 67.94%, and 74.76% for EC. Therefore, ELM-GWO can be applied as a robust approach for investigating leachate and groundwater quality parameters in different landfill sites.
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Mobasser, Farid, and Keyvan Hashtrudi-Zaad. "A Comparative Approach to Hand Force Estimation using Artificial Neural Networks." Biomedical Engineering and Computational Biology 4 (January 2012): BECB.S9335. http://dx.doi.org/10.4137/becb.s9335.
Повний текст джерелаАнотація:
In many applications that include direct human involvement such as control of prosthetic arms, athletic training, and studying muscle physiology, hand force is needed for control, modeling and monitoring purposes. The use of inexpensive and easily portable active electromyography (EMG) electrodes and position sensors would be advantageous in these applications compared to the use of force sensors which are often very expensive and require bulky frames. Among non-model-based estimation methods, Multilayer Perceptron Artificial Neural Networks (MLPANN) has widely been used to estimate muscle force or joint torque from different anatomical features in humans or animals. This paper investigates the use of Radial Basis Function (RBF) ANN and MLPANN for force estimation and experimentally compares the performance of the two methodologies for the same human anatomy, ie, hand force estimation, under an ensemble of operational conditions. In this unified study, the EMG signal readings from upper-arm muscles involved in elbow joint movement and elbow angular position and velocity are utilized as inputs to the ANNs. In addition, the use of the elbow angular acceleration signal as an input for the ANNs is also investigated.
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Alizamir, Meysam, Kaywan Othman Ahmed, Sungwon Kim, Salim Heddam, AliReza Docheshmeh Gorgij, and Sun Woo Chang. "Development of a robust daily soil temperature estimation in semi-arid continental climate using meteorological predictors based on computational intelligent paradigms." PLOS ONE 18, no. 12 (December 27, 2023): e0293751. http://dx.doi.org/10.1371/journal.pone.0293751.
Повний текст джерелаАнотація:
Changes in soil temperature (ST) play an important role in the main mechanisms within the soil, including biological and chemical activities. For instance, they affect the microbial community composition, the speed at which soil organic matter breaks down and becomes minerals. Moreover, the growth and physiological activity of plants are directly influenced by the ST. Additionally, ST indirectly affects plant growth by influencing the accessibility of nutrients in the soil. Therefore, designing an efficient tool for ST estimating at different depths is useful for soil studies by considering meteorological parameters as input parameters, maximal air temperature, minimal air temperature, maximal air relative humidity, minimal air relative humidity, precipitation, and wind speed. This investigation employed various statistical metrics to evaluate the efficacy of the implemented models. These metrics encompassed the correlation coefficient (r), root mean square error (RMSE), Nash-Sutcliffe (NS) efficiency, and mean absolute error (MAE). Hence, this study presented several artificial intelligence-based models, MLPANN, SVR, RFR, and GPR for building robust predictive tools for daily scale ST estimation at 05, 10, 20, 30, 50, and 100cm soil depths. The suggested models are evaluated at two meteorological stations (i.e., Sulaimani and Dukan) located in Kurdistan region, Iraq. Based on assessment of outcomes of this study, the suggested models exhibited exceptional predictive capabilities and comparison of the results showed that among the proposed frameworks, GPR yielded the best results for 05, 10, 20, and 100cm soil depths, with RMSE values of 1.814°C, 1.652°C, 1.773°C, and 2.891°C, respectively. Also, for 50cm soil depth, MLPANN performed the best with an RMSE of 2.289°C at Sulaimani station using the RMSE during the validation phase. Furthermore, GPR produced the most superior outcomes for 10cm, 30cm, and 50cm soil depths, with RMSE values of 1.753°C, 2.270°C, and 2.631°C, respectively. In addition, for 05cm soil depth, SVR achieved the highest level of performance with an RMSE of 1.950°C at Dukan station. The results obtained in this research confirmed that the suggested models have the potential to be effectively used as daily predictive tools at different stations and various depths.
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Hussain, Bydaa Ali, and Mohammed Sadoon Hathal. "Developing Arabic License Plate Recognition System Using Artificial Neural Network and Canny Edge Detection." Baghdad Science Journal 17, no. 3 (September 1, 2020): 0909. http://dx.doi.org/10.21123/bsj.2020.17.3.0909.
Повний текст джерелаАнотація:
In recent years, there has been expanding development in the vehicular part and the number of vehicles moving on the roads in all the sections of the country. Arabic vehicle number plate identification based on image processing is a dynamic area of this work; this technique is used for security purposes such as tracking of stolen cars and access control to restricted areas. The License Plate Recognition System (LPRS) exploits a digital camera to capture vehicle plate numbers is used as input to the proposed recognition system. Basically, the proposed system consists of three phases, vehicle license plate localization, character segmentation, and character recognition, the License Plate (LP) detection is presented using canny edge detection algorithm, Connect Component Analysis (CCA) have been exploited for segmenting characters. Finally, a Multi-Layer Perceptron Artificial Neural Network (MLPANN) model is utilized to identify and detect the vehicle license plate characters, and hence the results are displayed as a text on GUI. The proposed system successfully detects LP and recognizes multi-style Arabic characters with rates of 96% and 97.872% respectively under different conditions.
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Hussain, Bydaa Ali, and Mohammed Sadoon Hathal. "Development of Iraqi License Plate Recognition System based on Canny Edge Detection Method." Journal of Engineering 26, no. 7 (July 1, 2020): 115–26. http://dx.doi.org/10.31026/j.eng.2020.07.08.
Повний текст джерелаАнотація:
In recent years, there has been expanding development in the vehicular part and the number of vehicles moving on the road in all the sections of the country. Vehicle number plate identification based on image processing is a dynamic area of this work; this technique is used for security purposes such as tracking of stolen cars and access control to restricted areas. The License Plate Recognition System (LPRS) exploits a digital camera to capture vehicle plate numbers is used as input to the proposed recognition system. Basically, the developing system is consist of three phases, vehicle license plate localization, character segmentation, and character recognition, the License Plate (LP) detection is presented using canny Edge detection algorithm, Connect Component Analysis (CCA) have been exploited for segmenting characters. Finally, a Multi-Layer Perceptron Artificial Neural Network (MLPANN) model is utilized to recognize and detect the vehicle license plate characters, and hence the results are displayed as a text on GUI. The proposed system successfully identified and recognized multi_style Iraqi license plates using different image situations and it was evaluated based on different metrics performance, achieving an overall system performance of 91.99%. This results shows the effectiveness of the proposed method compared with other existing methods, whose average recognition rate is 86% and the average processing time of one image is 0.242s which proves the practicality of the proposed method.
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Prashanth, M., D. Madhu, K. Ramanarasimh, and R. Suresh. "Effect of Heat Input and Filling Ratio on Raise in Temperature of the Oscillating Heat Pipe with Different Working Fluids Using ANN Model." International Journal of Heat and Technology 40, no. 2 (April 30, 2022): 535–42. http://dx.doi.org/10.18280/ijht.400221.
Повний текст джерелаАнотація:
In the present article, feed forward multilayer perceptron neural network (FFMLPNN) model has been used to predict the rise in temperature in closed loop oscillating heat pipe filled with three different fluid i.e., Acetone, methanol and ethanol respectively. Experimental test was carried out for the inner diameter of 1.7mm copper tube for all the combinations of filling ratio, heat input and time taken to evaluate the performance of the OHP. Totally 2000 data sets have been used for Acetone and Methanol, 1500 data sets is used for ethanol in the present NN model. ANN model with FFMLPNN using three input parameter (Filling ratio, heat input and time taken) and rise in temperature has output parameter respectively. Levenberg-Marquardt algorithm with a 4-10 neurons has been used for the determination of optimal model. The 3-8-1 combinations predict the rise in temperature for ethanol and acetone whereas for methanol 3-7-1 is the optimal combinations was achieved. For all the combinations RMSE values are 0.3414, 0.1285 and 0.1237 (Training-70%), 0.3526, 0.1375, 0.1234 (testing-15%) and 0.3010, 0.1515, 0.1425 (validation-15%). The values for coefficient of determinations are 0.9941, 0.9975 and 0.9971 for methanol, acetone and ethanol was achieved. The results clearly indicated that the proposed MLPANN model can successfully predict the rise in temperature.
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Gebremariam, Gebrekiros Gebreyesus, J. Panda, and S. Indu. "Localization and Detection of Multiple Attacks in Wireless Sensor Networks Using Artificial Neural Network." Wireless Communications and Mobile Computing 2023 (January 10, 2023): 1–29. http://dx.doi.org/10.1155/2023/2744706.
Повний текст джерелаАнотація:
Security enhancement in wireless sensor networks (WSNs) is significant in different applications. The advancement of routing attack localization is a crucial security research scenario. Various routing attacks degrade the network performance by injecting malicious nodes into wireless sensor networks. Sybil attacks are the most prominent ones generating false nodes similar to the station node. This paper proposed detection and localization against multiple attacks using security localization based on an optimized multilayer perceptron artificial neural network (MLPANN). The proposed scheme has two major part localization techniques and machine learning techniques for detection and localization WSN DoS attacks. The proposed system is implemented using MATLAB simulation and processed with the IBM SPSS toolbox and Python. The dataset is classified into training and testing using the multilayer perceptron artificial neural network to detect ten classes of attacks, including denial-of-service (DoS) attacks. Using the UNSW-NB, WSN-DS, NSL-KDD, and CICIDS2018 benchmark datasets, the results reveal that the suggested system improved with an average detection accuracy of 100%, 99.65%, 98.95%, and 99.83% for various DoS attacks. In terms of localization precision, recall, accuracy, and f-score, the suggested system outperforms state-of-the-art alternatives. Finally, simulations are done to assess how well the suggested method for detecting and localizing harmful nodes performs in terms of security. This method provides a close approximation of the unknown node position with low localization error. The simulation findings show that the proposed system is effective for the detection and secure localization of malicious attacks for scalable and hierarchically distributed wireless sensor networks. This achieved a maximum localization error of 0.49% and average localization accuracy of 99.51% using a secure and scalable design and planning approach.
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SAHIN, CENK, SEYFETTIN NOYAN OGULATA, KEZBAN ASLAN, HACER BOZDEMIR, and RIZVAN EROL. "A NEURAL NETWORK-BASED CLASSIFICATION MODEL FOR PARTIAL EPILEPSY BY EEG SIGNALS." International Journal of Pattern Recognition and Artificial Intelligence 22, no. 05 (August 2008): 973–85. http://dx.doi.org/10.1142/s0218001408006594.
Повний текст джерелаАнотація:
Epilepsy is a disorder of cortical excitability and still an important medical problem. The correct diagnosis of a patient's epilepsy syndrome clarifies the choice of drug treatment and also allows an accurate assessment of prognosis in many cases. The aim of this study is to evaluate epileptic patients and classify subgroups of partial epilepsy by Multilayer Perceptron Neural Networks (MLPNNs). This is the first study to classify the partial epilepsy groups using the neural network according to EEG signals. 418 patients with epilepsy diagnoses according to International League against Epilepsy (ILAE, 1981) were included in this study. The epilepsy outpatients at the Neurology Department Clinic of Cukurova University Medical School between the years of 2002–2005 were examined and included in the study. The MLPNNs were trained by the parameters obtained from the EEG signals and clinical findings of the patients. Test results show that the MLPNN model is able to classify partial epilepsy with an accuracy of 91.5%. Moreover, new MLPNNs were constructed for determining significant variables on classification. The loss of consciousness in the course of seizure time variable caused the largest decrease in the classification accuracy when it was left out. In conclusion, we think that the classification performance of MLPNN model for partial epilepsy is satisfactory and this model may be used in clinical studies as a decision support tool to determine the partial epilepsy classification of the patients.
Дисертації з теми "MLPANN"
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Ó, Vivian Tragante do. "Análise por MLPA das regiões subteloméricas de pacientes com Holoprosencefalia." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/61/61132/tde-09012015-120002/.
Повний текст джерелаАнотація:
Introdução: A Holoprosencefalia (HPE) é uma malformação craniofacial decorrente de falhas no processo de formação do sistema nervoso durante o desenvolvimento embrionário. Sua etiologia é heterogênea e complexa, envolvendo fatores ambientais e genéticos. Estudos recentes sugerem que alterações subteloméricas possam ser um dos fatores etiológicos para o aparecimento da HPE. Objetivos: Investigar possíveis alterações (microdeleções/duplicações) nas regiões subteloméricas em indivíduos com diagnóstico de HPE. Metodologia: Avaliação genética de 25 amostras de DNA de indivíduos matriculados no HRAC-USP, com diagnóstico de HPE através da técnica de MLPA (Multiplex ligation-dependent probe amplification), segundo protocolo proposto por Schouten et al. (2002). Estes indivíduos já foram previamente triados para mutações/deleções nos genes candidatos à HPE (SHH, ZIC2, TGIF, GLI2 e PTCH1) através do sequenciamento direto de Sanger e da técnica de MLPA, sem resultado positivo para qualquer alteração. Resultados: Dentre os 25 indivíduos analisados, o fenótipo predominante foi a microforma da doença. Os principais achados clínicos da amostra estudada foram: hipotelorismo, microcefalia e fissura de lábio e palato (100%), nariz achatado (84%), presença de um incisivo central único (24%) e ponte nasal baixa (64%). Quatro pacientes apresentaram comprometimento no SNC (16%). Nenhum indivíduo apresentou quaisquer alterações, como microdeleções e/ou duplicações nos genes contidos nas regiões subteloméricas, através da análise pela técnica de MLPA. Dessa forma, estes permanecem sem diagnóstico genético definido. Discussão: A não detecção de alterações subteloméricas sugere que o fenótipo predominante na amostra, microforma, possa não apresentar alterações subteloméricas relacionadas ao aparecimento da doença. Entretanto, deve-se salientar que o universo amostral é relativamente pequeno, de forma que este possa não ser um exemplo fiel dos casos de microforma da HPE. Reforça-se, ainda, a grande variedade de fatores envolvidos no surgimento desta patologia, bem como o envolvimento de outros genes ainda não estabelecidos, além das causas ambientais, ainda não completamente elucidadas. Conclusões: Não foram encontradas alterações subteloméricas nos pacientes com diagnóstico de HPE estudados. A técnica de MLPA consiste em uma metodologia rápida, sensível, eficaz e de baixo custo, quando comparada a outras técnicas, sendo indicada para o uso em laboratórios de diagnóstico genético, uma vez que diversos estudos já mostraram que consiste em um método confiável de diagnóstico. Devido ao tamanho relativamente pequeno da amostra utilizada neste estudo, e os dados ainda inconsistentes da literatura atual, estudos adicionais são necessários para que seja possível a realização de um diagnóstico diferencial, explicando o amplo espectro fenotípico observado nesta doença, conforme sugerido pela hipótese dos múltiplos fatores.Introduction: Holoprosencephaly (HPE), a craniofacial malformation, results from flaws in formation process of the nervous system during embryonic development. The etiology is heterogeneous and complex, involving environmental and genetic factors. Recent studies suggest that subtelomeric aberrations could be an etiological factor to the onset of HPE. Objectives: Investigate possible changes (microdeletions/duplications) in subtelomeric regions in individuals diagnosed with HPE. Methodology: Genetic evaluation of 25 DNA samples from individuals enrolled at HRAC-USP, diagnosed with HPE (performed by Syndromology Division HRAC/USP) by MLPA technique, as proposed by Schouten et al (2002). Patients were previously screened for mutations/deletions in HPE candidate genes (SHH ZIC2, TGIF, GLI2 and PTCH1) by direct Sanger sequencing and MLPA technique, without any match. Analyses were performed at the Laboratory of Molecular Genetics, Hospital for Rehabilitation of Craniofacial Anomalies, HRAC-USP. Results: Among the 25 individuals analyzed, the predominant phenotype was HPE microform. The main clinical findings of the study sample were: hypotelorism, microcephaly, and cleft lip/palate (100%); flat nose (84%); presence of a single central incisor (24%) and low nasal bridge (64%). Four patients had CNS commitment (16%). No subtelomeric mutations were found in our sample, such as microdeletions/duplications of genes analyzed by MLPA technique. Thus, they remain without defined genetic diagnosis. Discussion: Subtelomeric changes were not found, suggesting that the predominant sample phenotype, microform HPE, could not be related with subtelomeric changes associated to the disease outbreak. However, it should be noted that the sample universe is relatively small, so this may not be a true example of HPE microform cases. It should also be reinforced the wide variety of factors involved in the onset of this pathology, as well as the involvement of other genes not yet established and environmental causes, not completely understood. Conclusions: No subtelomeric mutations were found in the HPE individuals studied. MLPA technique consists in a rapid, sensitive and cost effective methodology, when compared to other techniques being suitable for use in genetic diagnostic laboratories, since several studies have shown that consists of a reliable method of diagnosis. Due to the relatively small sample size used in this study, and even inconsistent data in literature, further studies are needed to make it possible to perform a differential diagnosis, explaining the wide phenotypic spectrum observed in this disease, as suggested by the multiple hit hypothesis.
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Lindberg, Magdalena. "Utökning av panelför multiplex RT-MLPA av singel celler för prostatacancer diagnostik." Thesis, KTH, Skolan för bioteknologi (BIO), 2015. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-173122.
Повний текст джерелаАнотація:
The most common cancer form among men i prostate cancer and measurement of circulating tumor cells (CTCs) in the blood is a useful tool to evaluating the responset o treatment and progress of disease. CTC’s are cells that have detached from the original tumor and have spread in the blood sstem. In this master thesis a panel of 16 genes at single cell levels are analyzed using Reverse Transciptase Multiplexed Ligation-dependent Probe Amplification (RT-MLPA). The method uses gene specific reverse transcription primers to generate and amplify cDNA which MLPA probes are hybridized to. Correctly hybridized probes are ligated and amplified so that relative expression profiles can be calculated. One additional MLPA probe was designed to add to the existing panel. The results show that the MLPA reaction generates products from all genes in the panel when performed on synthetic MLPA prbe targets with equal concentrations. Results from totatl RNA on cell lines show that the reverse transcription and amplification of cDNA need further optimizations. When the whole assay is working it will be possible to evaluate gene expression from CTC’s that can help us understand the progression and spread of prostate cancer in the body.Prostatacancer är den vanligaste cancerformen hos män. Att räkna antal cirkulerade tumörceller (CTCer) i blodet är ett bra verktyg för att undersöka hur sjukdomen fortlöper och hur den svarar på behandling. CTCer är enskilda celler som brutit sig loss från originaltrumören och sprids i kroppen genom blodsystemet. I det här examensarbetet analyseras en panel bestående av 16 gener med hjälp av Reverse Transcriptase Multiplexed Ligation-dependent Probe Amplification (RT-MLPA). Metoden bygger på att genspecifika primrar används för att syntetisera cDNA från mRNA. MLPA-prober hybridisear sedan till det amplifierade cDNAt. MLPA prober som hybridiserat korrekt ligeras och amplifieras och den relativa uttrycksnivåerna kan beräknas. Ytterligare en MLPA-probe designades för att passa in i den existerande blandningen av MLPA-prober. Resultaten viar att alla MLPA-prober ger produkter då en syntetisk DNA-templat blandning med lika koncentrationer används. Resultaten från total-RNA från cellinjer visar att omvandlingen och amplifieringen av mRNA till cDNA måste optimeras. När hela protokollet fungerar är det möjligt att undersöka genuttrycken i CTCer vilket kan underlätta förståelsen för utveckling och spridning av prostatacancer i kroppen.
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Nascimento, Juliana Minuncio. "Análise do perfil de mutações driver por MLPA em pacientes com Mielofibrose." reponame:Repositório Institucional da UnB, 2017. http://repositorio.unb.br/handle/10482/31249.
Повний текст джерелаАнотація:
Dissertação (mestrado)—Universidade de Brasília, Faculdade de Medicina, Programa de Pós-Graduação em Ciências Médicas, 2017.Submitted by Raquel Viana (raquelviana@bce.unb.br) on 2018-02-16T18:16:36Z No. of bitstreams: 1 2017_JulianaMinuncioNascimento.pdf: 2253571 bytes, checksum: b4527c088c7b8285e11c0f28fe5c865e (MD5)
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A Mielofibrose é a mais rara e severa das Neoplasias Mieloproliferativas Crônicas Philadelphia negativas. Caracteriza-se por fibrose medular, hematopoese extramedular e expressão anormal de citocinas inflamatórias, que resultam em citopenias, organomegalias, sintomas constitucionais, eventos trombohemorrágicos e evolução para Leucemia Aguda. Pode ocorrer de novo ou pós Policitemia Vera ou Trombocitemia Essencial, a partir da expansão clonal de uma célula tronco hematopoética desencadeada por uma mutação somática envolvendo os genes JAK2, CALR ou MPL, combinada a desregulação dos nichos hematopoéticos, a mutações e a anomalias citogenéticas adicionais. Este estudo visou caracterizar o perfil de mutações driver de portadores de Mielofibrose primária e secundária acompanhados em um serviço público terciário de Hematologia, e correlacionar este perfil aos desfechos clínicos dos doentes. A pesquisa das mutações JAK2 V617F (éxon 14) e éxon 12, CALR c.1092_1143del52 e c.1154_1155insTTGTC (éxon 9) e MPL W515K e W515L foi realizada em 31 indivíduos por meio da técnica de MLPA, método de análise de DNA que permite a pesquisa simultânea de diferentes mutações, em diferentes amostras. A mutação JAK2V617F foi encontrada em 48,4% dos pacientes, mutações indel do éxon 9 da CALR em 38,7% (em 66,7% destes a mutação del52, e em 33,3% a mutação insTTGTC), e a mutação MPL W515L em 3,2% dos pacientes. 9,7% dos pacientes eram triplo-negativos. Os pacientes com JAK2 mutada eram mais idosos, com menor grau de anemia e maior frequência de leucocitose, enquanto os portadores de mutações da CALR apresentavam menor frequência de leucocitose e plaquetopenia. Os indivíduos triplo-negativos apresentaram a menor mediana de idade ao diagnóstico (49,3 anos) e fenótipo de falência medular semelhante a Síndrome Mielodisplásica. A estratificação de risco por DIPSS foi semelhante à relatada em outros centros. O tempo mediano de acompanhamento foi de 32 meses (variando de 10 meses a 13 anos), sendo registrados fenômenos tromboembólicos em 19,3% e evolução para LMA em 6,4% dos pacientes. A taxa de mortalidade foi de 29%, e a sobrevida média após o diagnóstico foi de 68,3 meses. Os indivíduos com mutação da CALR apresentaram maior sobrevida média. A mediana de sobrevida de acordo com o DIPSS foi superior à prevista pelo modelo prognóstico, possivelmente pela maior frequência de mutações da CALR observada nesta população. Maior tempo de seguimento e inclusão de novos pacientes são necessários para melhor avaliação de desfechos e confirmação da maior prevalência de mutações da CALR. A pesquisa de mutações driver é essencial para sustentação diagnóstica, define subgrupos de doentes com características clínicas peculiares e, aliada a pesquisa de mutações colaborativas, tem impacto prognóstico. O complexo panorama genético envolvido na iniciação e progressão das NMP, especialmente a Mielofibrose, instiga a adoção de modelos integrativos de estratificação prognóstica. Neste cenário, o MLPA é uma potente ferramenta para estudo molecular, e um promissor aliado na caracterização das NMP.
Myelofibrosis is the rarest and most severe Philadelphia-negative myeloproliferative neoplasm and can present de novo or post Polycythemia Vera or Essential Thrombocythemia. It is characterized by bone marrow fibrosis, extramedullary hematopoiesis and abnormal expression of inflammatory cytokines, which result in cytopenias, organomegaly, constitutional symptoms, thrombohemorrhagic events and progression to Acute Leukemia. The disease arises from clonal expansion of a single hematopoietic stem cell (HSC), driven by a somatic mutation of JAK2, CALR or MPL genes combined with dysregulation of hematopoietic microenvironment, additional mutations and cytogenetic abnormalities. This study aimed to assess driver mutations status in patients with primary and secondary myelofibrosis accompanied at a tertiary Brazilian public hospital, and to correlate their mutational profile with clinical outcomes. The search for JAK2V617F, exon 12 JAK2, calreticulin exon 9 c.1092_1143del52 and c.1154_1155insTTGT, MPLW515K and MPLW515L mutations was performed in 31 subjects using MLPA technique, a method of DNA analysis that allows simultaneous appraisal of different mutations in multiple samples. JAK2V617F mutation was found in 48.4% of patients, indel CALR mutations in 38.7% of patients (of these, 66.7% harbored del52 bp, 33.3% harbored insTTGTC), MPL W515L in 3.2% of patients and 9.7% of patients were triple-negative. Patients with mutated JAK2 were older, with minor degree of anemia and more leukocytosis, whereas those with CALR mutations had less frequency of leukocytosis and thrombocytopenia. Triple-negative subjects displayed the lowest median age at diagnosis (49.3 years), and bone marrow failure phenotype, similar to Myelodysplastic Syndrome. Risk stratification provided by DIPSS was similar to other centers. Median follow-up time was 32 months (ranging from 10 months to 13 years). Thromboembolic phenomena were recorded in 19.3% of patients, and evolution to AML in 6.4% of patients. Mortality rate was 29%, and mean survival after diagnosis was 68.3 months. CALR mutated individuals presented higher average survival. Median survival according to DIPSS was higher than predicted by the prognostic model, possibly due to the higher frequency of CALR mutations reported. Longer follow-up and inclusion of new patients are necessary for better evaluation of outcomes and confirmation of the higher prevalence of CALR mutations. Driver mutations assessment is essential for diagnostic support, defines subgroups with peculiar clinical characteristics and, combined with collaborative mutations evaluation, has prognostic impact. The complex genetic landscape involved in initiation and progression of MPN, especially Myelofibrosis, instigates the adoption of integrative prognostic stratification models. In this scenario, MLPA is a powerful tool for molecular study, and a promising ally for MPN molecular characterization.
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Jun, M. S. "Governing Marine Protected Areas (MPAs) in California : analysis of the MLPA implementation process." Thesis, University College London (University of London), 2014. http://discovery.ucl.ac.uk/1427878/.
Повний текст джерелаАнотація:
Neoliberal governance strategies have been hegemonic in shaping global policy toward Marine Protected Areas (MPAs) over the past two decades. This impact has manifested itself in two key dimensions: the prominence given to public-private partnerships (PPPs) and the dependency upon civil society, particularly in the form of non-governmental organisations (NGOs). As a result, it is necessary to give primacy to the implication of PPPs and the role of NGOs in considering how best to govern MPAs. This is particularly the case in relation to efforts which seek the ‘right’ combination of ‘the market’, ‘the people’ and ‘the state’. This thesis investigates the California Marine Life Protection Act (MLPA) implementation process, and particularly the MLPA Initiative, which is widely publicised as a successful case of a science-based stakeholder-driven process through PPP. The thesis involves a thorough exploration of how an ideal combination could be achieved based on the Central Coast Study Region (CCSR) MLPA implementation process. Number of literary sources identified four key factors which have significantly contributed to the implementation of MLPA: 1) A strong legal mandate 2) Strong political will 3) A substantial level of stakeholder participation 4) Effective PPPs However, despite the widely publicised claims, research findings suggest that finding the ‘right’ combination for the MLPA implementation process remains a difficult task. The strong legal mandate, which has provided the foundation for the science used, constrained the stakeholder participation process. Indeed, it suggests that the terms ‘science-based’ and ‘stakeholder-driven’ could be to some extent, oxymorons, whilst strong political will could potentially compromise stakeholder participation. Effective PPPs for the MLPA Initiative represent a conundrum for PPP, since NGOs, including philanthropic foundations, increasingly exercise their influence on public policy to push through their agendas. Subsequently, PPP could potentially compromise the legitimacy of the process. Finally, the research findings suggest that the substantial level of stakeholder participation may not be a panacea for designating MPAs.
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Moura, Karla Cristina Vasconcelos. "Relação entre fatores genéticos envolvidos em vias metabólicas mitocondriais e a doença de Parkinson." Universidade do Estado do Rio de Janeiro, 2013. http://www.bdtd.uerj.br/tde_busca/arquivo.php?codArquivo=6826.
Повний текст джерелаАнотація:
Conselho Nacional de Desenvolvimento Científico e TecnológicoA doença de Parkinson (DP) é uma das desordens neurodegenerativas mais comuns associada ao envelhecimento, alcançando 2% aos 70 anos. É uma doença caracterizada pela degeneração progressiva de neurônios dopaminérgicos nigrais nos gânglios basais e pela presença de inclusões protéicas citoplasmáticas denominadas corpúsculos e neuritos de Lewy nos neurônios sobreviventes. A etiologia da DP é pouco conhecida, sendo considerada, na maioria dos casos, idiopática. Conhecimentos alcançados nos últimos 15 anos sobre a base genética da DP demonstram, claramente, que os fatores genéticos desempenham um importante papel na etiologia desta desordem. Neste trabalho, rastreamos mutações nos genes que codificam proteínas participantes de vias metabólicas mitocondriais (Parkin, PINK1 e DJ-1) em 136 pacientes brasileiros com manifestação precoce da DP, através do sequenciamento automático e da técnica de MLPA. Avaliamos a presença de variantes de sequência por meio do sequenciamento dos exons 1 a 12 do gene Parkin e dos exons 1 a 8 do gene PINK1. Em Parkin foram identificadas três mutações patogênicas ou potencialmente patogênicas, ambas em heterozigose: p.T240M, p.437L e p.S145N. Em PINK1 não encontramos variantes de ponto patogênicas. Através da técnica de MLPA investigamos alterações de dosagem nos genes Parkin, PINK1 e DJ-1. Identificamos cinco alterações no gene Parkin em quatro pacientes: uma duplicação heterozigota do exon 4 no paciente PAR2256, uma deleção heterozigota do exon 4 no probando PAR2099, uma deleção homozigota do exon 4 na paciente PAR3380 e um probando heterozigoto composto (PAR2396) com duas alterações, uma duplicação do exon 3 e uma deleção dos exons 5 e 6. No gene PINK1 identificamos uma deleção heterozigota do exon 1, que nunca foi descrita na literatura, em um paciente (PAR2083). Não encontramos alteração quantitativa no gene DJ-1. Neste estudo obtivemos uma frequência total de mutações patogênicas (pontuais e de dosagem) nos genes estudados de 7,3%, sendo 6,6% no gene Parkin e 0,7% no gene PINK1.
Parkinson's disease (PD) is one of the most common neurodegenerative disorders associated with aging, reaching 2% at age 70. It is a disease characterized by progressive degeneration of nigra dopaminergic neurons in the basal ganglia and the presence of cytoplasmic protein inclusions known as Lewy bodies and neurites in surviving neurons. The etiology of PD is poorly understood, being considered, in most cases, idiopathic. Knowledge achieved in the last 15 years about the genetic basis of PD clearly shows that genetic factors play an important role in the etiology of this disorder. In this study, we screened mutations in genes that encode proteins participating in mitochondrial metabolic pathways (Parkin, PINK1 and DJ-1) in 136 Brazilian patients with early onset PD, through automatic sequencing and MLPA technique. We evaluated the presence of sequence variants by means of sequencing of exons 1 to 12 of Parkin gene and exons 1 to 8 of PINK1 gene. In Parkin gene were identified three pathogenic or potentially pathogenic mutations, both in heterozygous state: p.T240M, p.437L e p.S145N. In PINK1 gene we did not find pathogenic point mutations. Through the MLPA technique we investigated dosage changes in Parkin, PINK1 and DJ-1 genes. We identified five exon rearrangements in Parkin gene in four patients: a heterozygous duplication of exon 4 in patient PAR2256, a heterozygous deletion of exon 4 in proband PAR2099, a homozygous deletion of exon 4 in patient PAR3380 and a compound heterozygote (PAR2396) with two changes, a duplication of exon 3 and a deletion of exons 5 and 6. In PINK1 gene we identified a heterozygous deletion of exon 1, which has never been described in literature, in one patient (PAR2083). We found no quantitative change in DJ-1 gene. In this study, we obtained an overall frequency of pathogenic mutations (sequence and dosage) in the genes studied of 7.3%, being 6.6% in Parkin gene and 0.7% in PINK1 gene.
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Suemasu, Cintia Natsumi. "Caracterização dos genótipos da talassemia alfa delecional por MLPA (Multiplex Ligation-Dependent Probe Amplification)." [s.n.], 2010. http://repositorio.unicamp.br/jspui/handle/REPOSIP/308947.
Повний текст джерелаАнотація:
Orientador: Maria de Fátima SonatiDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: A talassemia alfa (a) constitui um grupo de doenças hereditárias causadas pela redução ou ausência da síntese das cadeias 'alfa' da hemoglobina. Os genes responsáveis pela codificação dessas cadeias são duplicados ('alfa' 2 e 'alfa'1) e estão localizados em um agrupamento gênico ou cluster, situado próximo ao telômero do cromossomo 16 (16p13.3). As deleções são as principais causas da doença, podendo afetar um ou ambos os genes 'alfa' do cromossomo (formas 'alfa' + e 'alfa' 0, respectivamente). Nos últimos anos, várias técnicas foram desenvolvidas para identificar as deleções envolvendo o cluster dos genes a no cromossomo 16. A gap- PCR, o Southern blot e o FISH são comumente aplicados com esta finalidade; entretanto, muitas deleções ainda não são detectadas utilizando-se essas estratégias metodológicas. Em 2005, uma técnica simples e adequada a análises quantitativas rápidas, o Multiplex Ligation-dependent Probe Amplification (MLPA), foi adaptada ao diagnóstico das hemoglobinopatias. Esta técnica é baseada na ligação e na amplificação, em reação de PCR, de dois oligonucleotídeos (sondas) que se hibridizam de forma adjacente ao DNA alvo. Cada oligonucleotídeo é projetado para gerar um produto de comprimento único e, por possuírem terminações comuns, todas as sondas podem ser amplificadas utilizando um único par de iniciadores. Por fim, o uso de fluorocromos permite a separação dos fragmentos, que foram gerados pela amplificação das sondas, em sistema de seqüenciamento por capilaridade. No presente estudo nós utilizamos o MLPA para investigar as bases moleculares da talassemia alfa em nove pacientes não relacionados, cinco deles com Doença da Hb H, cujos diagnósticos moleculares não puderam ser esclarecidos pelas técnicas usuais, incluindo o seqüenciamento de DNA. No total, foram utilizadas 44 sondas, distribuídas desde o telômero ao gene MSLN, cobrindo uma região de cerca de 800 kb, na região 16p13.3. Oito diferentes deleções foram detectadas. Enquanto quatro delas comprometem uma região de grande extensão que inclui todo o locus a e seu elemento regulatório, com tamanhos mínimos de 240 kb, 470 kb, 500 kb e 720 kb, respectivamente (posições 97000-334571, 97000-570532, 97000-602492 and 97000- 816477 do UCSC Genome Browser, Fevereiro, 2009, respectivamente), quatro outras encontram-se limitadas às regiões que contém o elemento regulatório, deixando os genes a intactos, porém sem expressão. Essas últimas apresentam tamanhos mínimos de 0.4 kb, em um dos casos, e de 95 kb a 100 kb nos demais (posições entre 163464-163904, 97000- 193847, 97000-202417 e 97000-202417 do UCSC Genome Browser, Fevereiro, 2009, respectivamente). Em um dos pacientes com fenótipo talassêmico heterozigótico foi demonstrada a presença de uma quadruplicação de cerca de 230 kb de DNA, incluindo todo o cluster a e seu elemento regulatório (97000 a 231370 do UCSC Genome Browser, Fevereiro, 2009). Este estudo representa o primeiro no Brasil a empregar o método de MLPA na caracterização das bases moleculares da talassemia a. A ampla diversidade de alterações identificadas enfatiza a necessidade de se investigar todos os casos cujos valores hematológicos revelem hipocromia e microcitose, na ausência de deficiência de ferro e níveis normais de HbA2
Abstract: Alpha (a) thalassemia is an inherited hemoglobin disorder characterized by reduction or absence of the a-globin chain synthesis. These chains are encoded by duplicated genes ('alpha' 2 and 'alpha' 1) that lie in a gene cluster near the telomeric end of the short arm of chromosome 16 (16p13.3). Deletions are the major molecular cause of the disease and may affect one or both 'alfa genes in the chromosome (the 'alpha' + and 'alpha' 0 forms, respectively). In recent years several techniques have been developed to identify deletions involving the 'alpha' -globin cluster in chromosome 16. The molecular tests commonly used to identify these deletions are gap- PCR, Southern Blot analysis and FISH. However, several thalassemia deletions remain uncharacterized by these methods. In 2005 a simple technique suitable for rapid quantitative analysis ¾ multiplex ligationdependent probe amplification (MLPA) ¾ was adapted for the diagnosis of thalassemias. The approach is based on the ligation and PCR amplification of two adjacently hybridizing oligonucleotides (probes). Each oligonucleotide pair is designed to give a product of unique length, and by using common ends all the probes can be amplified with one primer pair. Finally, the use of a fluorescent label allows the probe to be separated in a capillary sequencing system. In this study we used MLPA to investigate the molecular basis of thalassemia in nine unrelated patients (five with Hb H disease) in whom the molecular causes of the disease could not be detected by sequence analysis or other conventional techniques. In total, 44 probe pairs were used, covering approximately 800 kb from the telomere to the MSLN gene in the 16p13.3 region. Eight different deletions were detected. While four of them affect a large region including the entire ? -globin gene locus and its upstream regulatory element, spanning genomic regions of at least 240 kb, 470 kb, 500 kb and 720 kb (positions 97000-334571, 97000-570532, 97000-602492 and 97000-816477 of the UCSC Genome Browser, February, 2009, respectively). The other four deletions were limited to a region that contains the upstream regulatory element; the ? -globin genes, although intact, are therefore not expressed. These deletions span a region of at least 0.4 kb in one case and from 95 kb to 100 kb in the other cases (positions 163464-163904, 97000- 193847, 97000-202417 and 97000-202417 of the UCSC Genome Browser, February, 2009, respectively). One carrier who was suspected of having heterozygous ? -thalassemia showed a segmental quadruplication spanning about 230 kb and including the a cluster and upstream regulatory element (97000-231370 do UCSC Genome Browser, February, 2009). This study is the first in Brazil to use the MLPA method to characterize thalassemias. The variety of rearrangements identified highlights the need to investigate all cases presenting with microcytosis and hypochromia without iron deficiency or elevated Hb A2 levels
Mestrado
Ciencias Biomedicas
Mestre em Ciências Médicas
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Cuevas, Sánchez Dolors. "Aplicació de tecnologies d’alt rendiment per a l’anàlisi d’alteracions moleculars i l’heterogeneïtat intratumoral en el càncer d’endometri." Doctoral thesis, Universitat de Lleida, 2020. http://hdl.handle.net/10803/670256.
Повний текст джерелаАнотація:
Els dos subtipus més freqüents de càncer d’endometri (CE) són el carcinoma endometrioide (CEE) i el carcinoma serós (CSE). El diagnòstic diferencial entre aquestes dues tipologies no sempre és fàcil, hi ha casos que presenten característiques histològiques molt dubtoses i ambigües que dificulten el correcte diagnòstic final. Aquests dos tipus tumorals presenten diferents perfils moleculars i un pronòstic diferencial, fent que el CSE sigui el subtipus més agressiu i amb el pronòstic més desfavorable. Actualment s’està posant en coneixement la importància que pot jugar l’heterogeneïtat intratumoral (HIT) en els comportaments més agressius dels tumors i en la resistència als tractaments, com és el cas del CSE. En els últims anys, la irrupció de les tecnologies d’alt rendiment com la NGS (Next Generation Sequencing) i la MLPA (Multiple Ligase-dependent Probe Amplification) han ofert l’opció d’ampliar el coneixement molecular del càncer d’una manera més precisa i resolutiva.
Per aquests motius, els dos primers objectius d’aquesta tesi s’han centrat en analitzar, en els dos tipus més freqüents de càncer d’endometri, diverses alteracions moleculars que ens permetin obtenir uns perfils genètics específics, i classificar d’una manera més precisa i objectiva aquests dos tipus tumorals. Per realitzar les anàlisis de les alteracions genètiques s’han utilitzat, l’aplicació de targeted sequencing (NGS) amb un panell personalitzat de 40 gens per determinar els gens alterats respecte a variants somàtiques, i la tècnica MLPA per determinar les alteracions somàtiques en el número de còpies (ASNC) de 106 gens. Finalment, el tercer objectiu s’ha focalitzat en conèixer el paper que juga l’heterogeneïtat intratumoral en el CSE. L’efecte de l’HIT ha estat estudiat a dos nivells d’alteració molecular, a nivell de l’alteració dels gens determinant variants somàtiques i de l’alteració en el número de còpies dels gens, mitjançant les dues tècniques anteriors.
Els resultats del primer objectiu d’aquesta tesi han demostrat la idoneïtat del nostre estudi personalitzat de NGS com a eina molecular addicional per confirmar la classificació histològica del CE. Aquesta estratègia sembla interessant com una eina per classificar tumors amb troballes microscòpiques inusuals i ambigües. Amb els resultats del segon objectiu s’ha descrit que de manera general el CSE presenta moltes més ASNC que el CEE. Tot i això, també s’ha mostrat que dins el CSE hi ha un percentatge de casos (42%) que presenten unes característiques genètiques que no es corresponen amb el fenotip, i que per tant, en aquests casos pot ser de gran ajuda l’ús del perfil genètic d’ASNC per realitzar una classificació més correcta. A més a més, s’ha suggerit l’ús de la combinació de la determinació de p53 per immunohistoquímica i del número de còpies de la CCNE1 per classificar els casos dins el grup serous-like. Per finalitzar, els resultats del tercer objectiu han mostrat que el CSE, per una banda, presenta unes alteracions moleculars clonals com les variants somàtiques al gen TP53 i guanys en els gens CCNE1 i PIK3CA, i per altra banda, que la seva heterogeneïtat intratumoral està caracteritzada principalment per les ASNC dels gens. A més a més, s’ha remarcat que un dels gens principalment afectats per aquesta heterogeneïtat és el gen ERBB2, que és una diana terapèutica àmpliament utilitzada.Los dos subtipos más frecuentes de cáncer de endometrio (CE) son el carcinoma endometrioide (CEE) y el carcinoma seroso (CSE). El diagnóstico diferencial entre estas dos tipologías no siempre es fácil, hay casos que presentan características histológicas muy dudosas y ambiguas que dificultan su correcto diagnóstico final. Estos dos tipos tumorales presentan perfiles moleculares y pronósticos diferenciales, haciendo que el CSE sea el subtipo más agresivo y con el pronóstico más desfavorable. Actualmente se está poniendo en conocimiento la importancia que puede jugar la heterogeneidad intratumoral (HIT) en los comportamientos más agresivos de los tumores y en la resistencia a los tratamientos, como es el caso del CSE. En los últimos años, la irrupción de las tecnologías de alto rendimiento como la NGS (Next Generation Sequencing) y la MLPA (Multiple Ligase-dependiente Probe Amplification) han ofrecido la opción de ampliar el conocimiento molecular del cáncer de una manera más precisa y resolutiva. Por estos motivos, los dos primeros objetivos de esta tesis se han centrado en analizar en los dos tipos más frecuentes de cáncer de endometrio varias alteraciones moleculares que nos permitan obtener unos perfiles genéticos específicos, y clasificar de una manera más precisa y objetiva estos dos tipos tumorales. Para realizar los análisis de las alteraciones genéticas se han utilizado, la aplicación de targeted sequencing (NGS) con un panel personalizado de 40 genes para determinar los genes alterados respecto a variantes somáticas, y la técnica MLPA para determinar las alteraciones somáticas en el número de copias (ASNC) de 106 genes. Finalmente, el tercer objetivo se ha focalizado en conocer el papel que juega la heterogeneidad intratumoral en el CSE. El efecto de la HIT ha sido estudiado a dos niveles de alteración molecular, a nivel de la alteración de los genes analizando las variantes somáticas y la alteración en el número de copias de los genes, mediante las dos técnicas anteriores. Los resultados del primer objetivo de esta tesis han demostrado la idoneidad de nuestro estudio personalizado de NGS como herramienta molecular adicional para confirmar la clasificación histológica del CE. Esta estrategia parece interesante como una herramienta para clasificar tumores con hallazgos microscópicos inusuales y ambiguos. Con los resultados del segundo objetivo se ha descrito que de manera general el CSE presenta muchas más ASNC que el CEE. Sin embargo, también se ha mostrado que dentro el CSE hay un porcentaje de casos (42%) que presentan unas características genéticas que no corresponden con el fenotipo, por lo que en estos casos puede ser de gran ayuda el uso del perfil genético de ASNC para realizar una clasificación más correcta. Además, se ha sugerido el uso de la combinación de la determinación de p53 por inmunohistoquímica y del número de copias de la CCNE1 para clasificar los casos dentro del grupo serous-like. Para finalizar, los resultados del tercer objetivo han mostrado que el CSE, por una parte, presenta unas alteraciones moleculares clonales como las variantes somáticas en el gen TP53 y ganancias en los genes CCNE1 y PIK3CA, y por otra parte, que su heterogeneidad intratumoral está caracterizada principalmente por las ASNC de los genes. Además, se ha remarcado que uno de los genes principalmente afectados por esta heterogeneidad es el gen ERBB2, que es una diana terapéutica ampliamente utilizada.
The two most common subtypes of endometrial cancer (CE) are endometrioid carcinoma (CEE) and serous carcinoma (CSE). Differential diagnosis between these two types is not always easy, there are cases with very doubtful and ambiguous histological features that make it difficult their final diagnostic. These two tumor types have different molecular profiles and differential prognosis, making the CSE the most aggressive subtype and with the most unfavorable prognosis. At present, the importance of intratumoral heterogeneity (HIT) can be played out in the most aggressive behaviors of tumors and in resistance to treatment, as in the case of CSE. In recent years, the advent of high-performance technologies such as NGS (Next Generation Sequencing) and MLPA (Multiple Ligase-dependent Probe Amplification) have offered the option of extending molecular knowledge of cancer in a way more accurate and resolute. For these reasons, the first two objectives of this dissertation have been to analyze in the two most common types of endometrial cancer, various molecular alterations that allow us to obtain specific genetic profiles, and to classify more precisely and objective these tumor types. To preform genetic alteration analyzes have been used the application of targeted sequencing (NGS) with a personalized panel of 40 genes to determine the altered genes with respect to somatic variants, and the MLPA technique to determine somatic alterations in the number of copies (ASNC) of 106 genes. Finally, the third objective was focused on understanding the role that intratumoral heterogeneity plays in CSE. The effect of HIT has been studied at two levels of molecular alteration, at the level of the genes determining somatic variants and the number of copies of the genes, using the two previous techniques. The results of the first objective of this thesis have shown the suitability of our personalized NGS study as an additional molecular tool to confirm the histological classification of CE. This strategy seems interesting as a tool for classifying tumors with unusual and ambiguous microscopic findings. The results of the second objective have described that in general the CSE has many more ASNCs than the EEC. However, it has also been shown that in the CSE there is a percentage of cases (42%) that have genetic characteristics that do not correspond to the phenotype, and therefore in these cases the use may be very helpful of the ASNC gene profile for more accurate classification. In addition, the combination of p53 determination by immunohistochemistry and the number of CCNE1 copies has been suggested to classify cases within the group serous-like. Finally, the results of the third objective have shown that CSE, on the one hand, has clonal molecular alterations such as the somatic variants in the TP53 gene and gains in the CCNE1 and PIK3CA genes, and on the other hand, that its intratumoral heterogeneity is characterized mainly by the ASNC of the genes. It has also been emphasized that one of the genes that is most affected by this heterogeneity is the ERBB2 gene, which is a widely used therapeutic target.
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Murray, Michael S. "The genetic analysis of mlpA, a gene encoding a myosin-like protein in Physarum polycephalum." Thesis, University of Leicester, 1990. http://hdl.handle.net/2381/34453.
Повний текст джерелаАнотація:
A cDNA clone encoding a myosin-like protein has been detected by screening a ?gtll expression library with antibodies specific to a myosin-like protein extracted from Physarum amoebae. The clone, mlpA, encodes a polypeptide which is expressed in both cell stages in Physarum, and which shows significant homology to the myosin heavy chain rod domain of Dictyostelium discoideum. Genetic analysis has determined that there is a single gene that encodes mlpA in Physarum, and that this gene is not related to a mutation, hts-23, analysed in earlier work.
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Uehara, Daniela Tiaki. "Pesquisa de microrrearranjos em genes candidatos a surdez sindrômica e não-sindrômica." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/41/41131/tde-22022011-111240/.
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A complexidade da fisiologia da audição resulta da participação e interação de produtos de grande número de genes, razão pela qual a surdez hereditária exibe enorme heterogeneidade genética. Estudos moleculares nas duas últimas décadas permitiram a identificação de vários genes responsáveis por surdez; entretanto, muitos ainda restam ser identificados. A maioria dos estudos de mapeamento de genes de surdez até então conduzidos privilegiou estratégias que buscavam mutações de ponto. Outros mecanismos mutacionais, como deleções e duplicações, foram pouco investigados. Portanto, a contribuição das CNVs (Copy Number Variations) na surdez hereditária é pouco conhecida. O objetivo desse trabalho foi identificar novos genes que possam ter papel na etiologia da surdez sindrômica ou não-sindrômica por meio da investigação de microdeleções e microduplicações em pacientes com perda auditiva. Selecionamos 25 genes candidatos (CTTN, FGF3, FGF19, FOXC1, FOXF2, FOXQ1, IMMP2L, KIF5C, LRRN3, MAP1A, MYLK4, PPP3CA, SHANK2, SLC5A7, STRC, TMC1, TMC2, TMC3, TMC4, TMC5, TMC6, TMC7, TMC8, TPCN2 e TUBB2A) para a triagem de microrrearranjos por meio da técnica de MLPA (Multiplex Ligation-dependent Probe Amplification). Os genes candidatos foram selecionados a partir de rearranjos detectados em um estudo prévio realizado por meio de array-CGH (array-based Comparative Genomic Hybridization) em indivíduos com surdez sindrômica estudados em nosso laboratório, e também a partir de dados da literatura. Nossa casuística foi composta por 163 indivíduos, dos quais 74 são pacientes com surdez associada a outros sinais (sindrômicos), a maioria casos isolados, e 89 são pacientes com surdez não-sindrômica, propósitos de famílias em que segrega surdez de herança autossômica dominante ou recessiva. Desenhamos uma sonda sintética intragênica de MLPA para cada um dos genes candidatos. Foram detectadas seis deleções em TMC6 (3,7%), seis deleções e uma duplicação em STRC (4,3%) e uma duplicação em IMMP2L (0,6%). A triagem de alterações nesses três genes em 189 indivíduos fenotipicamente normais revelou quatro deleções em TMC6 (2,1%), oito deleções e três duplicações em STRC (5,8%) e três deleções em IMMP2L (1,6%). Todas as alterações em TMC6, tanto nos casos de surdez como nos controles, eram na realidade artefatos devidos a problemas de hibridação da sonda correspondente. No gene STRC, previamente já relacionado à surdez, os rearranjos nos indivíduos afetados devem se tratar de polimorfismos sem efeito fenotípico por serem muito frequentes na população. Contudo, é possível que haja nesses pacientes mutações adicionais que não puderam ser rastreadas e que poderiam contribuir ao fenótipo, em combinação com o rearranjo detectado, como já descrito em um caso da literatura. A duplicação em IMMP2L em uma paciente com surdez não-sindrômica, herdada da mãe igualmente afetada, mostrou-se a mais provavelmente relacionada ao fenótipo, pois o estudo complementar por meio de array-CGH revelou que o rearranjo inclui uma duplicação parcial da porção 3 de outro gene, DOCK4. O produto desse gene possui uma isoforma que se localiza nos estereocílios das células ciliadas e se liga a uma importante proteína relacionada à audição, a harmonina. Portanto, nossa hipótese é a de que a duplicação seja a causa da surdez na família e que DOCK4 seja um novo gene responsável por surdez. A associação de IMMP2L com surdez é menos provável devido ao grande número de CNVs não patogênicas já descritas que incluem partes desse gene. Estudos complementares são necessários para mapear a duplicação com mais precisão. Além disso, o rastreamento de mutações em DOCK4 em outras famílias com surdez pode vir a confirmar o possível papel desse gene na etiologia da surdez.Several genes contribute to the complexity of physiology of hearing. Consequently, hereditary deafness is extremely heterogeneous from the genetic point of view. In the last two decades, several genes responsible for hereditary hearing loss have been identified, but a large number of genes remains to be found, as evidenced by the unexplained cases of inherited deafness. The search for point mutations in candidate genes after mapping based on linkage studies has been the main strategy in the identification of such genes. Other mutation mechanisms, such as deletions and duplications, have been rarely investigated, and the contribution of DNA copy number variants (CNVs) to hearing loss is not well known. This study aimed at identifying novel genes, which might play a role in the etiology of syndromic and non-syndromic deafness, through the search of gene microdeletions and microduplications. We selected 25 candidate genes (CTTN, FGF3, FGF19, FOXC1, FOXF2, FOXQ1, IMMP2L, KIF5C, LRRN3, MAP1A, MYLK4, PPP3CA, SHANK2, SLC5A7, STRC, TMC1, TMC2, TMC3, TMC4, TMC5, TMC6, TMC7, TMC8, TPCN2 and TUBB2A) based on their involvement in microimbalances detected by Array-based Comparative Genomic Hybridization (aCGH) in a previous study of a Brazilian sample of individuals with syndromic hearing loss from our laboratory and others reported in the literature. We studied 163 subjects, 74 of them presenting syndromic deafness, the majority were isolated cases, and 89 being probands of families in which nonsyndromic deafness had an autosomal dominant or recessive mode of inheritance. Gene deletions or duplications were screened by Multiplex Ligant-dependent Probe Amplification (MLPA) using one synthetic intragenic probe designed for each candidate gene. We detected six deletions in TMC6 (3,7%), six deletions and one duplication in STRC (4,3%), and one duplication in IMMP2L (0,6%). The screening of imbalances in these genes in a control sample of 189 hearing individuals revealed four deletions in TMC6 (2,1%), eight deletions and three duplications in STRC (5,8%) and three deletions in IMMP2L (1,6%). The imbalances found in TMC6, both in affected and control individuals, were in fact artifacts due to problems in the hybridization of the corresponding probe. As to the STRC gene, previously related to deafness, the imbalances are more likely to be 4 polymorphisms with no phenotypic effect. However, the possibility remains that additional undetected mutations in affected individuals contribute to their phenotype, in combination with the microrearrangement, as already reported in the literature. The duplication in IMMP2L in a non-syndromic patient, and also present in her affected mother, is most likely causative of deafness, since a complementary study performed with aCGH revealed that the rearrangement included a partial duplication of the 3 end of another gene, DOCK4. An isoform of the DOCK4 protein localizes to the stereocilia in the inner ear and interacts with harmonin, a protein already known to be involved in hearing. We hypothesize that this duplication may be the cause of deafness in the family and, this being the case, DOCK4 appears as a novel deafness gene. The causal association between IMMP2L and deafness is less plausible, because of the large number of reported non-pathogenic CNVs that include parts of this gene. Further studies are required to precisely map this duplication. In addition, the screening of mutations in DOCK4 in other families with hearing impairment is required to evaluate its possible role in the etiology of deafness.
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Goveia, Rebeca Mota. "Análise de deleção/ duplicação nos genes BRCA1 e BRCA2 em pacientes de Goiás-Brasil com suspeita da síndrome do câncer de mama e ovário hereditário." Universidade Federal de Goiás, 2018. http://repositorio.bc.ufg.br/tede/handle/tede/8723.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
Introduction: Breast cancer is the second most common cancer in the world, and the most common among women, only 5% to 10% are hereditary and half of them are caused by hereditary breast and ovarian cancer (HBOC) , caused by variations in the BRCA1 and BRCA2 genes. Objectives: The present study aimed to identify the prevalence of deletions and duplications in BRCA1 and BRCA2 genes in breast cancer patients in Goiás, Brazil. Materials and methods: We evaluated 46 breast cancer patients who met National Comprehensive Cancer Network (NCCN) criteria for HBOC syndrome screening. A 4 ml blood sample was collected for DNA extraction using commercial kit and the MLPA (Multiplex Ligation Dependent Probe Amplification) technique was performed using the SALSA MLPA P002 BRCA1 and SALSA MLPA P045 BRCA2 / CHECK2 kits. Results and discussion: The majority of the patients were female (97.83%) and the mean age of the patients was 37.52 years. In this group, 43.47% of the patients were younger than 35 years at the time of diagnosis and 35% of them were diagnosed with triple negative tumors. The most common molecular subtype was luminal A (46.2%) followed by triple negative tumors (28.20%). No patient was found with rearrangements in BRCA1. In the BRCA2 gene, one patient (2.12%) presented a false positive result for the heterozygous deletion of exon 27, which may have been caused by the presence of a small change in the probe binding region. Conclusion: This was the first study performed to analyze large deletions and duplications in patients from the central-western region of Brazil. We can conclude that the frequency of large deletions and duplications in the BRCA1 and BRCA2 genes in the Goian population is low.
Introdução: O câncer de mama é o segundo tipo de câncer mais freqüente no mundo, e o mais comum entre as mulheres, apenas 5% a 10% são hereditários e metade deles são causados pela síndrome do câncer de mama e ovário hereditário (HBOC), causada por variações nos genes BRCA1 e BRCA2. Objetivos: O presente estudo teve como objetivo identificar a prevalência de deleções e duplicações nos genes BRCA1 e BRCA2 em pacientes com câncer de mama no estado de Goiás, Brasil. Materiais e métodos: Avaliamos 46 pacientes com câncer de mama que atenderam aos critérios do National Comprehensive Cancer Network (NCCN) para pesquisa da síndrome HBOC. Foi coletada uma amostra de sangue de 4 ml para extração de DNA usando kit comercial e a técnica MLPA (Multiplex Ligation Dependent Probe Amplification) foi realizada usando os kits SALSA MLPA P002 BRCA1 e SALSA MLPA P045 BRCA2 / CHECK2. Resultados e discussão: A maioria dos pacientes era do sexo feminino (97.83%) e a idade média dos pacientes era de 37,52 anos. Neste grupo 43.47% dos pacientes possuíam idade inferior a 35 anos no momento do diagnóstico sendo que 35% destes foram diagnosticados com tumores triplo negativo. O subtipo molecular mais comum foi o luminal A (46.2%) seguido de tumores triplo negativos (28.20%). Nenhum paciente foi encontrado com rearranjos no gene BRCA1. No gene BRCA2, um paciente (2,12%) apresentou um resultado falso positivo para a deleção em heterozigoze do éxon 27, fato que pode ter sido ocasionado pela presença de uma pequena alteração na região de ligação da sonda. Conclusão: Este foi o primeiro estudo realizado para análise de grandes deleções e duplicações em pacientes da região centro-oeste do Brasil. Podemos concluir que a frequência de grandes deleções e duplicações nos genes BRCA1 e BRCA2 na população goiana é baixa.
Книги з теми "MLPANN"
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Manners Look Pretty on You: Mlpon. Independently Published, 2020.
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Team, The Examelot. MLPAO MLA/T Certification Exam: Practice Test 1. Independently Published, 2020.
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Team, The Examelot. MLPAO MLA/T Certification Exam: Practice Test 2. Independently Published, 2020.
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MLPAO MLA/T Certification Exam: Practice Test 5. Independently Published, 2021.
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Team, The Examelot. MLPAO MLA/T Certification Exam: Practice Test 4. Independently Published, 2021.
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Arnemann, J. "MLPA." In Springer Reference Medizin, 1669–70. Berlin, Heidelberg: Springer Berlin Heidelberg, 2019. http://dx.doi.org/10.1007/978-3-662-48986-4_3533.
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Arnemann, J. "MLPA." In Lexikon der Medizinischen Laboratoriumsdiagnostik, 1–2. Berlin, Heidelberg: Springer Berlin Heidelberg, 2018. http://dx.doi.org/10.1007/978-3-662-49054-9_3533-1.
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Mohapatra, Sushree Shataroopa, Manoj Kumar Maharana, Abhilash Pradhan, P. K. Panigrahi, and R. C. Prusty. "Islanding Detection in Distributed Generation System Using MLPNN and ELPID Methods." In Smart Technologies for Power and Green Energy, 211–21. Singapore: Springer Nature Singapore, 2022. http://dx.doi.org/10.1007/978-981-19-2764-5_18.
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Jha, Panchanand, Bibhuti Bhusan Biswal, and Om Prakash Sahu. "Intelligent Computation of Inverse Kinematics of a 5-dof Manipulator Using MLPNN." In Advances in Autonomous Robotics Systems, 243–50. Cham: Springer International Publishing, 2014. http://dx.doi.org/10.1007/978-3-319-10401-0_22.
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Ohnesorg, Thomas, Erin Turbitt, and Stefan J. White. "The Many Faces of MLPA." In Methods in Molecular Biology, 193–205. Totowa, NJ: Humana Press, 2010. http://dx.doi.org/10.1007/978-1-60761-944-4_13.
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Berbegall, Ana Pilar, Eva Villamón, Samuel Navarro, and Rosa Noguera. "Multiplex Ligation-dependent Probe Amplification (MLPA)." In Guidelines for Molecular Analysis in Archive Tissues, 215–24. Berlin, Heidelberg: Springer Berlin Heidelberg, 2011. http://dx.doi.org/10.1007/978-3-642-17890-0_33.
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Ohnesorg, Thomas, Stefanie Eggers, and Stefan J. White. "Detecting DNaseI-Hypersensitivity Sites with MLPA." In Methods in Molecular Biology, 201–10. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-292-2_12.
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Mahesh, Vijayalakshmi G. V., Alex Noel Joseph Raj, and P. Arulmozhivarman. "Thermal IR Face Recognition Using Zernike Moments and Multi Layer Perceptron Neural Network (MLPNN) Classifier." In Advances in Intelligent Systems and Computing, 213–22. Cham: Springer International Publishing, 2017. http://dx.doi.org/10.1007/978-3-319-60618-7_21.
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Schouten, Jan, and Robert-Jan Galjaard. "MLPA for Prenatal Diagnosis of Commonly Occurring Aneuploidies." In Prenatal Diagnosis, 111–22. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-066-9_8.
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Moelans, Cathy B., Lilit Atanesyan, Suvi P. Savola, and Paul J. van Diest. "Methylation-Specific Multiplex Ligation-Dependent Probe Amplification (MS-MLPA)." In Methods in Molecular Biology, 537–49. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-7481-8_27.
Повний текст джерелаТези доповідей конференцій з теми "MLPANN"
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Tahamipour-Z., S. Mohammad, Iman Kardan, Hadi Kalani, and Alireza Akbarzadeh. "A PSO-MLPANN Hybrid Approach for Estimation of Human Joint Torques from sEMG Signals." In 2020 8th Iranian Joint Congress on Fuzzy and intelligent Systems (CFIS). IEEE, 2020. http://dx.doi.org/10.1109/cfis49607.2020.9238724.
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Miano, Carlo M., Massimiliano Gobbi, and Giampiero Mastinu. "A Tutorial on Present and Future Global Approximation Issues With Application to a Vehicle Design Problem." In ASME 2001 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2001. http://dx.doi.org/10.1115/imece2001/de-23264.
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Abstract A tutorial is presented on Global Approximation, a mathematical procedure useful for the optimisation of complex systems. The state-of-the art numerical algorithms that can be exploited within a Global Approximation approach are reviewed. Some original numerical procedures are also proposed, such as using Low Discrepancy Sequences to train Multilayer Perceptron Neural Network (MLPNN) or Radial Basis Function Neural Network (RBFNN). The final aim of the research is to find a mathematical algorithm for Global Approximation which requires a minimum of expertise to be effectively used. A number of candidate algorithms for such a kind of Global Approximation are comparatively tested with reference to a vehicle design optimisation problem. The result is that the RBFNN can be as efficient as the MLPNN, which is much harder to train, when used to find a satisfactory approximation of the relationships among design variables and objective functions which define the road holding and handling performance of a race car. It is envisaged that in the future Global Approximation could be used to derive the relationship between Pareto-optimal design variable values and Pareto-optimal objective function values.
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Verma, Alka, and J. N. Mathur. "An Examination on Intelligent Islanding Classification with MLPNN." In 2022 Fourth International Conference on Emerging Research in Electronics, Computer Science and Technology (ICERECT). IEEE, 2022. http://dx.doi.org/10.1109/icerect56837.2022.10060600.
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Frimpong, Emmanuel Asuming, Philip Yaw Okyere, and Johnson Asumadu. "On-line determination of transient stability status using MLPNN." In 2017 IEEE PES/IAS PowerAfrica. IEEE, 2017. http://dx.doi.org/10.1109/powerafrica.2017.7991194.
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Katiyar, Gauri, and Shabana Mehfuz. "MLPNN based handwritten character recognition using combined feature extraction." In 2015 International Conference on Computing, Communication & Automation (ICCCA). IEEE, 2015. http://dx.doi.org/10.1109/ccaa.2015.7148550.
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Ruslan, Fazlina Ahmat, Abd Manan Samad, Mazidah Tajuddin, and Ramli Adnan. "Flood prediction modeling using improved MLPNN structure: Case study Kuala Lumpur." In 2015 IEEE Conference on Systems, Process and Control (ICSPC). IEEE, 2015. http://dx.doi.org/10.1109/spc.2015.7473567.
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Tagougui, Najiba, Houcine Boubaker, Monji Kherallah, and Adel M. Alimi. "A hybrid MLPNN/HMM recognition system for online Arabic Handwritten script." In 2013 World Congress on Computer and Information Technology (WCCIT). IEEE, 2013. http://dx.doi.org/10.1109/wccit.2013.6618744.
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Chekhmane, G., and R. Benali. "EEG signals analysis using SVM and MLPNN classifiers for epilepsy detection." In 2022 5th International Symposium on Informatics and its Applications (ISIA). IEEE, 2022. http://dx.doi.org/10.1109/isia55826.2022.9993577.
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Rojatkar, D. V., K. D. Chinchkhede, and G. G. Sarate. "Handwritten Devnagari consonants recognition using MLPNN with five fold cross validation." In 2013 International Conference on Circuits, Power and Computing Technologies (ICCPCT). IEEE, 2013. http://dx.doi.org/10.1109/iccpct.2013.6528992.
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Ferrah, Imene, Ahmed Khaled Chaou, and Madjid Teguar. "Discharges Pattern Recognition on Uniformly Polluted Glass Surface Using an MLPNN Classifier." In 2021 IEEE Electrical Insulation Conference (EIC). IEEE, 2021. http://dx.doi.org/10.1109/eic49891.2021.9612333.
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