Academic literature on the topic 'Molecular Diagnosi'
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Journal articles on the topic "Molecular Diagnosi"
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Pietravalle, Andrea, Paola Cavicchioli, Enrica Donadel, Marta Lusiani, Tommaso Malusa, Giuliana Rossi, Giovanna Contreas, and Michela Chirico. "Diagnosi precoce di diabete neonatale permanente conseguente a mutazione del gene KCNJ11." Medico e Bambino pagine elettroniche 25, no. 2 (February 28, 2022): 39–42. http://dx.doi.org/10.53126/mebxxv039.
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Neonatal diabetes mellitus (NDM) is a rare condition characterized by onset of persistent hyperglycaemia with-in the first six months of life. Heterozygous mutations in KCNJ11 gene account for about half of the cases of per-manent form of NDM and are associated with a wide range of neurocognitive disabilities. In suspected NDM, immediate molecular genetic testing is recommended because most of the cases due to mutations in KCNJ11 gene are responsive to oral sulfonylurea (SU) therapy with excellent glycaemic control at long term follow up. The present report describes a case of early detection of permanent NDM, due to KCNJ11 gene mutation, which has been successfully treated with SU oral therapy.
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Kutty, Dr A. V. M. "Molecular Diagnosis: A boon to healthcare." JOURNAL OF CLINICAL AND BIOMEDICAL SCIENCES 10, no. 3 (September 15, 2020): 74–75. http://dx.doi.org/10.58739/jcbs/v10i3.4.
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Diagnostic procedures are one of the essen-tial components in healthcare system. These tests provide cardinal information to enable clinicians to make accurate medical diagnosis, decide on manage-ment and treatment of diseases. The field of molecu-lar diagnostics owe a great deal to the developments in molecular biology which reached newer vistas in the early part of twenty first century. The applicabil-ity of molecular diagnostics have gathered momen-tum subsequent to the accomplishments of the hu-man genome project.
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Zhao, Sen, Yuanqiang Zhang, Weisheng Chen, Weiyu Li, Shengru Wang, Lianlei Wang, Yanxue Zhao, et al. "Diagnostic yield and clinical impact of exome sequencing in early-onset scoliosis (EOS)." Journal of Medical Genetics 58, no. 1 (May 7, 2020): 41–47. http://dx.doi.org/10.1136/jmedgenet-2019-106823.
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BackgroundEarly-onset scoliosis (EOS), defined by an onset age of scoliosis less than 10 years, conveys significant health risk to affected children. Identification of the molecular aetiology underlying patients with EOS could provide valuable information for both clinical management and prenatal screening.MethodsIn this study, we consecutively recruited a cohort of 447 Chinese patients with operative EOS. We performed exome sequencing (ES) screening on these individuals and their available family members (totaling 670 subjects). Another cohort of 13 patients with idiopathic early-onset scoliosis (IEOS) from the USA who underwent ES was also recruited.ResultsAfter ES data processing and variant interpretation, we detected molecular diagnostic variants in 92 out of 447 (20.6%) Chinese patients with EOS, including 8 patients with molecular confirmation of their clinical diagnosis and 84 patients with molecular diagnoses of previously unrecognised diseases underlying scoliosis. One out of 13 patients with IEOS from the US cohort was molecularly diagnosed. The age at presentation, the number of organ systems involved and the Cobb angle were the three top features predictive of a molecular diagnosis.ConclusionES enabled the molecular diagnosis/classification of patients with EOS. Specific clinical features/feature pairs are able to indicate the likelihood of gaining a molecular diagnosis through ES.
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Burboa Meza, Cinthya Y., Alexandra Zazueta Avitia, David Ramírez Alvarado, Miguel A. Segura Castruita, Paola A. Palmeros Suarez, and Juan F. Gómez-Leyva. "DIAGNÓSTICO COMPARATIVO DE BRUCELOSIS MEDIANTE MÉTODOS SEROLÓGICOS Y MOLECULARES." e-CUCBA 8, no. 16 (May 31, 2021): 50–55. http://dx.doi.org/10.32870/ecucba.vi16.198.
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Brucellosis is an infectious disease that limits livestock development and greatly affects the livestock economy, being considered one of the most important and widely distributed zoonoses worldwide. Early diagnosis of this disease is an essential tool in its control and eradication. The methods recognized by NOM-041-ZOO-1995, such as the card test and complement fixation, present limitations in the diagnosis, compared to the PCR molecular technique. In the present work, a comparative diagnosis of Brucella spp. was performed by PCR amplification of the gene coding for a protein located in the outer membrane (Omp2a) of Brucella spp. and serological tests in blood, milk and cheese samples from goats and cattle. The results showed a higher sensitivity in the detection by PCR technique, while the card test and complement fixation showed inconsistencies due to the occurrence of false positives and negatives. Based on the results, it is suggested to include the PCR technique in the Mexican Official Standard as an objective alternative in the routine diagnosis of brucellosis.
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Greco, F. Anthony. "Cancer of Unknown Primary Site: Improved Patient Management with Molecular and Immunohistochemical Diagnosis." American Society of Clinical Oncology Educational Book, no. 33 (May 2013): 175–81. http://dx.doi.org/10.14694/edbook_am.2013.33.175.
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Cancer of unknown primary site (CUP) is a common heterogeneous clinicopathologic syndrome, but investigations and publications regarding these patients are rare. For the last 20 years, empiric “broad-spectrum” chemotherapy has been the standard therapy for the majority of these patients. More recently, improved immunocytochemistry and advent of gene-expression profiling have provided the diagnostic tools necessary to accurately define the tissue of origin in most patients. Molecular profiling assays complement standard pathologic diagnosis, and a recently reported large prospective study demonstrated an improvement in outcome for patients treated with site-specific therapy directed by the molecular assay diagnoses compared with empiric chemotherapy. Survival in molecularly diagnosed patients was as expected for those particular tumor types. The evaluation of patients has become more standardized. The empiric-chemotherapy era is ending and customized therapies based on accurate tissue of origin diagnoses have arrived. Eventually the recognition of the molecular aberrations responsible for the growth and metastasis of solid tumors, regardless of the tissue of origin, will lead to more precise and effective therapy for patients with advanced cancers.
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Fujita, Naohide, Osamu Akiyama, and Akihide Kondo. "PATH-08. THE IMPORTANCE OF RE-DIAGNOSIS OF TUMORS PREVIOUSLY CLASSIFIED AS CENTRAL NERVOUS SYSTEM PRIMITIVE NEUROECTODERMAL TUMORS." Neuro-Oncology 22, Supplement_3 (December 1, 2020): iii426. http://dx.doi.org/10.1093/neuonc/noaa222.644.
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Abstract BACKGROUND The recent molecular analyses have revealed that central nervous system primitive neuroectodermal tumors (CNS PNETs) those having clusters of small round tumor cells are genetically different tumors. However, the concepts of CNS PNET are complicated, and it is difficult to diagnose them appropriately in clinical field. To overcome this difficulty, we reviewed previous studies associated with CNS PNETs, and carried out several approaches, those are relatively easy access to use in clinics, for our 8 samples of embryonal brain tumors diagnosed CNS PNETs in our institution, initially. METHODS We used in combination with immunohistochemistry (IHC), Sanger sequence, Pyrosequence, polymerase chain reaction (PCR), real time PCR and copy number analysis referring recent reports. RESULTS In terms of the diagnosis three out of 8 cases were changed based on the results in this study from previous diagnoses. CONCLUSION In this review, it seemed that either the histopathological evaluation or molecular analyses would be not enough to make accurate diagnosis of CNS embryonal brain tumors, and it is essential to combine both of them including recent comprehensive analysis methods.
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Luk, Ho-Ming. "Angelman-Like Syndrome: A Genetic Approach to Diagnosis with Illustrative Cases." Case Reports in Genetics 2016 (2016): 1–6. http://dx.doi.org/10.1155/2016/9790169.
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Epigenetic abnormalities in 15q11-13 imprinted region andUBE3Amutation are the two major mechanisms for molecularly confirmed Angelman Syndrome. However, there is 10% of clinically diagnosed Angelman Syndrome remaining test negative. With the advancement of genomic technology like array comparative genomic hybridization and next generation sequencing methods, it is found that some patients of these test negative Angelman-like Syndromes actually have alternative diagnoses. Accurate molecular diagnosis is paramount for genetic counseling and subsequent management. Despite overlapping phenotypes between Angelman and Angelman-like Syndrome, there are some subtle but distinct features which could differentiate them clinically. It would provide important clue during the diagnostic process for clinicians.
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ERDOĞAN, Emrah, Merve YÜRÜK, Eda SİVCAN, Serkan KARACA, Orhan YILDIZ, and İzzet ŞAHİN. "Plasmodium ovale Sıtması ve Moleküler Tanısı: Relaps Olabilir mi?" Mikrobiyoloji Bulteni 53, no. 1 (January 15, 2019): 106–13. http://dx.doi.org/10.5578/mb.67713.
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Pohanka, M., R. Chlibek, K. Kuca, H. Bandouchova, and J. Pikula. "Diagnosis of tularemia using biochemical, immunochemical and molecular methods: a review." Veterinární Medicína 56, No. 9 (October 6, 2011): 453–61. http://dx.doi.org/10.17221/3207-vetmed.
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Tularemia, an infection caused by the intracellular gram-negative bacterium Francisella tularensis, is accompanied by high mortality and occurs throughout the Northern Hemisphere. The causative agent is also considered one of the most important biological warfare agents. As well as its taxonomy and epidemiology, the basic immunochemical, biochemical, and molecular approaches for disease diagnosis are outlined in this review. Aspects of immune responses during tularemia and damage to specific organs are discussed with regards to the predictive value of standard biomarkers. Bacterial burden is also considered as a limitation for polymerase-chain-reaction-based diagnosis.
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Eszlinger, Markus, Kurt Werner Schmid, and Ralf Paschke. "Clinical implications of molecular studies for the diagnosis of thyroid cancer." HORMONES 9, no. 1 (January 15, 2010): 51–56. http://dx.doi.org/10.14310/horm.2002.1253.
Full textDissertations / Theses on the topic "Molecular Diagnosi"
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D'AVERSA, Elisabetta. "Innovative approaches for molecular diagnosis of genetic diseases." Doctoral thesis, Università degli studi di Ferrara, 2019. http://hdl.handle.net/11392/2488091.
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The diagnosis of genetic diseases in fetal age is obtained using invasive procedures, but these hide a high risk of miscarriage. Recently, non-invasive pre-natal diagnosis, based on circulating cell-free fetal DNA (ccffDNA) analysis starting from a simple maternal peripheral blood sampling, has become increasingly important. The fetal sex determination is necessary for predicting the risk of X-linked disorders.
Against this background, the principal purpose of the first part of the research was the development of rapid and sensitive non-invasive diagnostic methods for the fetal sex determination, in particular at early gestational ages.
In order to detect the SRY gene on Y chromosome, in ccffDNA samples extracted from maternal plasma, the quantitative Real Time PCR (qRT-PCR) was first employed. The obtained results confirmed that the fetal sex determination can correctly performed from the 9th gestational week, as shown in an already reported study. The innovative technology digital droplet PCR (ddPCR) was applied to non-invasive sex diagnosis at early gestational ages: after optimizing the experimental conditions, ccffDNA samples at early gestational stages (12-4.5 weeks) were analysed, and for all of them the fetal gender was correctly determined achieving 100% accuracy.
β thalassemia is an autosomal recessive inherited disease associated with the absence (β0) or reduction (β+) of adult hemoglobin β chains. β thalassemia major is the most severe form of the disease, in fact, patients are unable to survive into adulthood without a therapeutic transfusion plan associated with iron chelation. The only definitive treatment is the bone marrow transplantation, which hides transplant-related complications. So, innovative therapeutic approaches are been investigated in order to employ a personalized therapy. For this reason, the detection of the specific pathogenic molecular alteration is crucial for the employment of the correct targeted and personalized therapy.
Therefore, the aim of the second part of the research was the development of rapid, sensitive and cost-effective pre- and post-natal diagnostic approaches for the identification of the four most common mutations causing β thalassemia in the Mediterranean area (β039, β+IVSI-110, β0IVSI-1, β+IVSI-6).
The first technique employed, for post-natal diagnosis, was BiacoreTM system: after the immobilization and the validation of a normal and a mutated probes on the instrument chip, the diagnosis was performed from single-stranded PCR products, obtained from genomic DNA of healthy subjects and heterozygous and homozygous patients. For all the specimens, it was possible to correctly discriminate the genotype. Another post-natal approach employed the qRT-PCR based on genotyping assays. After the optimization and the validation, the assays allowed the correct molecular diagnosis. The same approach, based on genotyping assays, was applied to non-invasive pre-natal screening of the paternally inherited mutations. The ccffDNA samples were pre-amplified and analysed showing that the developed genotyping assays could be efficiently employed for non-invasive pre-natal diagnosis of paternally inherited β thalassemia mutations, at least until the 9th gestational week.
In order to extend the diagnosis to earlier pregnancy and to maternally or both maternally and paternally inherited mutations, the ddPCR technology had been proposed as molecular approach. β039 and β+IVSI-110 genotyping assays were optimized, validated and employed for the analysis. For all the samples, in which the mutation was paternally inherited, the fetal genotype was correctly determined, also at 5th gestational week. For the samples in which the mutation was maternally or both parents inherited, two diagnostic ranges of allelic ratio values were identified. They were statistically distinct and not overlapping, allowing the correctly determination of fetal genotype.La diagnosi di patologie genetiche fetali viene effettuata a partire da materiale biologico prelevato tramite tecniche invasive, le quali presentano un elevato rischio di aborto. La diagnosi prenatale non invasiva, effettuata a partire da ccffDNA (DNA circolante fetale), isolato da sangue materno, negli ultimi anni, ha avuto una crescita esponenziale. La diagnosi del sesso, in età prenatale, permette di definire il rischio di malattie legate al cromosoma X. La prima parte della tesi ha come obbiettivo, quindi, lo sviluppo di approcci diagnostici molecolari non invasivi, rapidi e sensibili per la determinazione del sesso fetale, in particolare a settimane di gestazione precoci. Per verificare la presenza del gene SRY, sul cromosoma Y, nel ccffDNA, estratto da plasma materno, è stata impiegata, in primis, la Real Time PCR. I risultati hanno confermato il limite di rilevabilità della tecnica (9 settimane) individuato in uno studio preliminare del gruppo di ricerca. Per settimane di gestazione precoci è stata impiegata la tecnologia innovativa digital droplet PCR (ddPCR): dopo aver ottimizzato le condizioni sperimentali, sono stati analizzati campioni a settimane di gestazione comprese fra le 12 e le 4.5, e per tutti la tecnica si è dimostrata accurata al 100% nel determinare correttamente il sesso fetale. La β talassemia è una patologia autosomica recessiva associata all’assenza o riduzione delle catene β globiniche dell’emoglobina. I soggetti che presentano la forma più severa della patologia non possono sopravvivere senza trasfusioni associate a ferrochelanti. L’unico vero trattamento definitivo è il trapianto di midollo, il quale però è associato a forti rischi. Sono stati studiati, quindi, approcci terapeutici innovativi con l’obbiettivo di una terapia personalizzata. L’identificazione della specifica alterazione molecolare è, quindi, di fondamentale importanza al fine di scegliere la corretta strategia terapeutica. La seconda parte della ricerca ha, quindi, come obbiettivo lo sviluppo di approcci molecolari diagnostici pre- e postnatali rapidi, sensibili e poco costosi per l’identificazione delle più frequenti mutazioni talassemiche nel Mediterraneo (β039, β+IVSI-110, β+IVSI-6, β0IVSI-1). Per la diagnosi postnatale, è stata impiegata, in primis, la tecnologia BiacoreTM: una sonda mutata e normale sono state immobilizzate sul chip, validate e, successivamente, è stato estratto il DNA genomico di soggetti sani e pazienti omozigoti ed eterozigoti per le mutazioni considerate, ed effettuata l’analisi. Per tutti i campioni analizzati è stato possibile discriminare il diverso genotipo. Un secondo approccio ha previsto il disegno di saggi di genotipizzazione da utilizzare in Real Time PCR. Ottimizzati e validati, i saggi hanno permesso una corretta diagnosi di tutti i pazienti considerati. Lo stesso approccio, basato sui saggi di genotipizzazione, è stato utilizzato per la diagnosi prenatale delle mutazioni ereditate per via paterna. I campioni di ccffDNA sono stati preamplificati e analizzati, mostrando come questo approccio sia stato in grado di identificare il genotipo fetale a partire dalla nona settimana di gestazione. Per poter estendere la diagnosi prenatale a settimane di gestazione precoci e a mutazioni ereditate sia per via materna che da entrambi i genitori è stata utilizzata la ddPCR come approccio molecolare. I saggi di genotipizzazione per le mutazioni β039 e β+IVSI-110 sono stati ottimizzati, validati e utilizzati per l’analisi del ccffDNA. Per tutti i campioni in cui la mutazione è ereditata per via paterna, il genotipo fetale è stato correttamente individuato fino alla quinta settimana di gestazione. Per i campioni in cui solo la madre o entrambi i genitori sono portatori, è stato possibile individuare due intervalli, sulla base del rapporto fra allele mutato e normale, statisticamente separati, necessari per considerare il feto non portatore o eterozigote.
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Soler, Aznar Maria. "Nanoplasmonic biosensors for clinical diagnosis at the point of care." Doctoral thesis, Universitat Autònoma de Barcelona, 2015. http://hdl.handle.net/10803/298172.
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Aquesta Tesi Doctoral se centra en el desenvolupament de noves metodologies analítiques en biosensors òptics com a solucions alternatives per a la diagnosi o la monitorització terapèutica de diferents malalties, com ara l’al·lèrgia, la celiaquia o el càncer. En particular, es proposa l’ús de biosensors nanoplasmònics per a la detecció de biomarcardors presents en fluids humans de manera ràpida, sensible i que no requereixi d’amplificació de senyal o de l’ús d’etiquetes. Tant el ja ben establert biosensor de Ressonància de Plasmó Superficial (SPR) com un innovador biosensor nanoplasmonic basat en nanodiscs d’or han estat avaluats per a la seva aplicació real en l’àrea clínica.
Les distintes metodologies biosensores presentades estan basades en l’ús d’anticossos, tant com a elements de bioreconeixement o com a biomarcadors específics de malalties. Primer, es presenta un estudi en profunditat de dues estratègies d’immobilització orientada d’anticossos per tal d’obtenir immunoassaigs en format directe de biomarcadors proteics en fluids biològics. En segon lloc, es proposa una nova estratègia immunosensora per a la detecció de pèptids derivats del gluten directament en orina com a tècnica ràpida i no invasiva per al control dietètic de pacients celíacs. A més, s’han desenvolupat dues metodologies utilitzant el biosensor nanoplasmònic per a detectar anticossos circulants en sang com a biomarcadors de malalties. Per una banda, s’ha dissenyat una estratègia alternativa per a la diagnosi d’al·lèrgia als medicaments (en particular a l’antibiòtic amoxicil·lina) basada en uns receptors dendrimèrics per a la detecció directa d’anticossos tipus IgE en sèrum. Finalment, s’ha avaluat una nova estratègia biosensora per a quantificar específicament autoanticossos tumorals per a la diagnosi precoç de càncer colorectal.
El treball d’aquesta Tesi combina l’experiència del grup de recerca en el disseny i fabricació de tecnologia biosensora avançada i innovadora amb el desenvolupament de tècniques bioanalítiques i de química de superfície per tal de superar els reptes actuals relacionats amb el cost i el temps requerit per a les anàlisis clíniques. A més, l’àmplia experiència del grup de recerca en transferència tecnològica i les col·laboracions establertes durant la tesi doctoral amb empreses com Biomedal S.L. o Protein Alternatives S.L. obren oportunitats interesants de cara a facilitar el procés de transferència tecnològica per a la implementació real de biosensors tipus Point-of-Care.This Doctoral Thesis focuses on the development of novel analytical methodologies in optical biosensors as alternative solutions for diagnosis or therapy monitoring of relevant diseases, such as allergy, celiac disease or cancer. In particular, we propose the use of nanoplasmonic biosensors for a rapid, sensitive and label-free detection of biomarkers present in human fluids. Both the well-known Surface Plasmon Resonance (SPR) biosensor and an innovative nanoplasmonic biosensor based on gold nanodisks surfaces have been evaluated for their real application in the clinical field. The different biosensor methodologies make use of antibodies, either as biorecognition elements in immunoassays or as specific disease biomarkers for diagnostics. First, an in-depth study of two site-directed antibody immobilization strategies is presented for the direct immunoassay of protein biomarkers in biological fluids. In second place, a novel immunosensing strategy is proposed for the detection of gluten-derivative peptides in urine as a rapid and non-invasive technique for dietary control in celiac patients. On the other hand, two assays have been developed employing the nanoplasmonic biosensor to detect blood circulating antibodies as disease biomarkers. First, we have designed an alternative approach for drug allergy diagnosis (in particular for amoxicillin) based on dendrimer-based receptors, which enable the detection IgE antibodies directly in serum. And second, a new biosensing strategy is assessed to quantify specific tumor-related autoantibodies for the early diagnosis of colorectal cancer. The work in this Thesis combines the wide knowledge of the research group in the design and fabrication of powerful biosensor technology with the development of surface activation chemistry and bioanalytical techniques to overcome current challenges related to costly and time-consuming clinical analysis. Besides, the strong experience of our research group in technological transfer and the established collaborations during this doctoral work with companies as Biomedal S.L. or Protein Alternatives S.L. open up interesting opportunities to facilitate the technology-transfer process for the real implementation of Point-of-Care biosensors.
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GRIONI, ANDREA. "Application of modern data science to genomics and clinical research." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2020. http://hdl.handle.net/10281/279991.
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Dopo che il progetto sul genoma umano è stato completato nell'aprile del 2003, il flusso
continuo di nuovi database e dati di sequenziamento ha iniziato a trasformare il campo della
genomica in scienza basata sui dati. La bioinformatica analizza i dati sperimentali grezzi con
l'obiettivo di ottenere informazioni che descrivono le condizioni biologiche misurate, fornendo
così un potente strumento per studiare specifici meccanismi molecolari e genetici. Questa
conoscenza deve essere combinata con la genomica per decifrare le interrelazioni tra geni,
elementi regolatori, vie metaboliche e interazioni proteiche. L'apprendimento profondo,
conosciuto come Deep Learning, e’ una sottodisciplina dell'apprendimento automatico, è stato
recentemente applicato al campo della genomica, portando a risultati notevoli. I due obiettivi
principali di questo lavoro sono: lo sviluppo e le applicazioni di strumenti bioinformatici che
consentano lo studio delle basi genetiche della leucemia linfoblastica acuta e l'uso di tecniche
di apprendimento profondo per l'identificazione di piccoli elementi di RNA non codificanti del
genoma umano. Questa tesi fornisce al lettore una panoramica completa della recente
evoluzione della genomica come campo interdisciplinare di ricerca strettamente connesso con
l'informatica e l'analisi dei dati.After the completion of the human genome project in April 2003, the continuous flow of sequencing data and the development of new databases began to transform the field of genomics into data-driven science. Bioinformatics analyses raw experimental data with the aim to obtain information describing biological processes, thus providing a powerful tool to investigate specific molecular and genetic mechanisms. This domain knowledge in combination with genomics allows to decipher the interrelationships between genes, regulatory elements, metabolic pathways, and protein interactions. Deep learning, a subdiscipline of machine learning, has been recently applied to the field of genomics, leading to remarkable results. The two main objectives of this study were: the development and application of bioinformatic tools for the study of the genetic basis of acute lymphoblastic leukaemia, and the usage of deep learning techniques for the identification of small non-coding RNA elements in the human genome. This dissertation provides a comprehensive overview of the recent evolution of genomics as an interdisciplinary field of research strongly associated with computer science and data analysis.
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Shirahata, Mitsuaki. "Gene Expression-Based Molecular Diagnostic System for Malignant Gliomas Is Superior to Histological Diagnosis." Kyoto University, 2008. http://hdl.handle.net/2433/124241.
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Ramiro, Juliana. "Detecção molecular de fungos fitopatogênicos associados às sementes de soja." Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-28042015-143631/.
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Fungos fitopatogênicos veiculados por sementes de soja podem causar sérios prejuízos à cultura, bem como danos diretos reduzindo o poder germinativo, vigor e emergência das sementes. A detecção e identificação precisa de fitopatógenos é uma das estratégias mais importantes para iniciar as medidas preventivas ou curativas no controle de doenças de plantas. Para se evitar a introdução e propagação de patógenos em áreas onde ainda não ocorrem, uma atenção especial deve ser tomada na detecção de agentes patogênicos em sementes. Os métodos tradicionalmente utilizados para detectar e identificar fungos em sementes são, muitas vezes, demorados, laboriosos e exigem um conhecimento extenso de taxonomia clássica. Métodos moleculares têm sido utilizados para detectar, identificar e quantificar uma longa lista de fungos fitopatogênicos. A técnica da reação em cadeia da polimerase em tempo real (qPCR) é atualmente considerada a mais eficiente para a detecção de fitopatógenos, não exigindo conhecimentos taxonômicos especializados para interpretar seus resultados. Considerando a importância e as implicações da diagnose rápida e bem sucedida de agentes patogênicos em sementes de soja, este estudo teve como objetivo estabelecer uma metodologia para elevar a eficiência de detecção dos fungos fitopatogênicos Sclerotinia sclerotiorum, Colletotrichum truncatum, Phomopsis spp. e Corynespora cassiicola, encontrados com maior frequência em sementes de soja, por meio de qPCR. Iniciadores e sondas de hidrólise TaqMan® foram projetados para os diferentes patógenos e avaliados quanto à sua especificidade e sensibilidade. Curvas padrões, baseando-se em diluições seriadas dos DNAs alvos de iniciadores e sondas específicas, foram estabelecidas para a quantificação desses patógenos. Amostras de sementes de soja naturalmente infectadas, provenientes dos Estados de Goiás, Minas Gerais e Paraná, foram submetidas a testes de detecção por meio de qPCR e métodos tradicionais, para fins de comparação. De todos os iniciadores e sondas projetados, apenas os de Phomopsis spp. e S. sclerotiorum apresentaram-se específicos e sensíveis, viabilizando sua utilização na detecção desses patógenos. Por meio dos testes tradicionais de detecção, Phomopsis spp. apresentou incidência máxima de 2,75% em uma amostra de Minas Gerais e S. sclerotiorum não foi detectado em nenhuma das amostras avaliadas. O método de qPCR proporcionou a detecção de Phomopsis spp. em todas as amostras testadas, alcançando o nível de incidência máximo de 6,75%. em amostra de Minas Gerais. S. sclerotiorum não foi detectado em nenhuma das amostras avaliadas pelo método de qPCR. Comparando-se os métodos de detecção testados, a qPCR foi mais sensível na detecção de Phomopsis spp. em sementes de soja.Seed-born pathogenic fungi in soybean can cause serious damage to the crop, as well as direct damage by reducing seeds germination, vigor and emergence. The detection and accurate identification of plant pathogens is one of the most important strategies to initiate preventive and curative measures in the management of plant diseases. Particular attention should be taken in the detection of pathogens in seeds in order to avoid introduction and spread of pathogens in areas where they do not occur. The traditionally used methods for detection and identification of seed-born pathogenic fungi are often time consuming, laborious and require extensive knowledge of classical taxonomy. Molecular methods have been used to detect, identify and quantify a long list of plant pathogenic fungi. Quantitative real time polymerase chain reaction (qPCR) is currently considered the most efficient technique for the detection of pathogens because it does not require specialized taxonomical knowledge to interpret its results. Given the importance and implications of rapid and successful diagnosis of seed-born pathogenic fungi in soybean, this study aimed to establish a qPCR methodology to increase the detection efficiency of plant pathogenic fungi Sclerotinia sclerotiorum, Colletotrichum truncatum, Phomopsis spp. and Corynespora cassiicola, the most commonly occurring seed-born pathogenic fungi. Primers and TaqMan® hydrolysis probes were designed for these four pathogens and tested for specificity and sensitivity. Standard curves were established to quantify these pathogens, based on serial dilutions of the target DNA and specific primers and probes. Samples of naturally infected soybean seed from the states of Goiás, Minas Gerais and Paraná were subjected to detection tests using qPCR and traditional methods, for comparison purposes. From all primers and probes designed, only those for Phomopsis spp. and S. sclerotiorum showed up specificity and sensitivity, enabling their use to detect of these pathogens. Detection by traditional tests, resulted in a maximum Phomopsis spp. incidence of 2.75% in a sample from Minas Gerais and S. sclerotiorum was not detected in any of the samples. The detection and quantification of these pathogens by qPCR revealed the presence of Phomopsis spp. in all tested samples, the highest incidence level of 6.75% in a sample from Minas Gerais. S. sclerotiorum was not detected in any sample assessed by qPCR method. In comparison with traditional methods, qPCR was more sensitive in detecting Phomopsis spp. in soybean seeds.
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Wang, Lei. "Molecular Probes for Pancreatic Cancer Imaging." PDXScholar, 2016. http://pdxscholar.library.pdx.edu/open_access_etds/3108.
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Pancreatic ductal adenocarcinoma (PDAC) has the poorest five-year survival rate of any cancer. Currently, there are no effective diagnostics or chemotherapeutics. Surgical resection is the only curative therapy. However, most patients experience recurrence due largely to challenges in assessing tumor margin status in the operating room. Molecular probes that selectively highlight pancreatic cancer tissue, having the potential to improve PDAC margin assessment intraoperatively, are urgently needed. In this work, a series of red and near-infrared fluorescent probes is reported. Two were found to distribute to normal pancreas following systemic administration. One selectively accumulates in genetically modified mouse models of PDAC, providing cancer-specific fluorescence. In contrast to the small molecule probes reported previously, it possesses inherent affinity for PDAC cells and tissue, and thus does not require conjugation to targeting agents. Moreover, the probe exhibits intracellular accumulation and enables visualization of four levels of structure including the whole organ, tissue, individual cells and subcellular organelles. It can thus promote new strategies for precision image-guided surgery, pancreatic cancer detection, the monitoring of therapeutic outcomes and basic research.
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CATARINO, Aricléia de Moraes. "Detecção de vírus da videira por RT-PCR em tempo real e por extensão de primers alelo-específicos e caracterização molecular de isolados do Nordeste Brasileiro." Universidade Federal Rural de Pernambuco, 2015. http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5995.
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The grapevine (Vitis spp.) belongs to the family of Vitaceae, being the botanical species V. vinifera L. and V. labrusca L. the most and widely cultivated, due to their products consumed as fresh fruits, jam, juices and wines. Despite the high economical importance, several factors may severely affect this crop, including the diseases caused by viruses. This study aimed to verify the incidence of virus present in commercial vineyards of two producing areas in Northeastern Brazil and evaluate the efficiency of some molecular methods for detecting and identifying viral species associated with grapevine. Materials showing or not symptoms were collected from grapevine genotypes in vineyards of Pernambuco, Paraiba, Bahia and Rio Grande do Sul, Brazil, and Locorotondo, Province of Bari, of the Puglia Region, Italy. The first part of the work was conducted at the Virology Laboratory of the Embrapa Uva e Vinho, RS, Brazil. For the identification of viral agents in the samples collected in Brazil, the extraction of total RNA was performed, cDNAs were obtained and tested by real time RT-PCR, using primers and probes specific for the following viruses: Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 2, 3 e 4 (GLRaV-2, -3 e -4), Grapevine fleck virus (GFkV), Grapevine rupestris vein feathering virus (GRVFV) and Grapevine fanleaf virus (GFLV). DNA fragments, products of the RT-qPCR, corresponding to the CP gene of each virus were eluted, linked to pGEM-T Easy vector (Promega) and used to transform bacteria. The plasmid DNA was extracted from transformed bacterial colonies, confirming the presence of the cloned fragments, which were sequenced. The grapevine material collected in Locorotondo was processed at the Istituto per la Protezione Sostenibile delle Piante (CNR-IPSP) and at the Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi “Aldo Moro”. In order to detect in multiplex test the most relevant viruses involved in the aetiology of fanleaf degeneration and the complexes of leafroll and rugose wood of grapevine, amplification techniques based on Allele Specific Primer Extension (ASPE) were tested by using the obtained cDNAs. The results showed that the techniques aggregate some advantages, such as reduction in time and relative simplicity of implementation, completely eliminating the use of toxic reagents, such as the ethidium bromide. The use of multiplex facilitates amplification of multiple targets in a single reaction, reducing the time and cost of the analyzes.
A videira (Vitis spp.) pertence à família Vitaceae, sendo as espécies botânicas V. vinifera L. e V. labrusca L. cultivadas em maior escala devido seus produtos, consumidos na forma de frutos in natura, geleias, sucos e vinhos. Apesar da grande importância econômica, vários fatores podem comprometer a produção desta cultura, incluindo as doenças causadas por vírus. O presente trabalho teve como objetivos verificar a incidência de vírus presentes em vinhedos comerciais de duas áreas produtoras do Nordeste do Brasil e avaliar a eficiência de alguns métodos moleculares para detecção e identificação de espécies virais associadas à videira. Amostras, apresentando ou não sintomas, foram coletados de genótipos de videira em propriedades situadas em Pernambuco, Paraíba, Bahia e Rio Grande do Sul, Brasil, e em Locorotondo, Província de Bari, Região da Puglia, Itália. A primeira parte do trabalho foi conduzida no Laboratório de Virologia da Embrapa Uva e Vinho, RS, Brasil. Visando a identificação dos agentes virais, nas amostras coletadas no Brasil, foram realizadas as extrações do RNA total, obtidos os cDNAs e testados por PCR em Tempo Real, empregando-se primers e sondas, específicos para os seguintes vírus: Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine leafroll-associated virus 2, 3 e 4 (GLRaV-2, -3 e -4), Grapevine fleck virus (GFkV), Grapevine rupestris vein feathering virus (GRVFV) e Grapevine fanleaf virus (GFLV). Fragmentos de DNA, produtos da RTq-PCR, correspondentes ao gene da CP de cada vírus foram eluídos, ligados ao vetor pGEM-T Easy (Promega) e utilizados na transformação de bactéria. Foi extraído o DNA plasmidial das colônias bacterianas transformadas, confirmando-se a presença dos fragmentos clonados, os quais foram sequenciados. A segunda parte, realizada com o material coletado em Locorotondo, foi processada no Istituto per la Protezione Sostenibile delle Piante (CNR-IPSP) e no Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi “Aldo Moro”. Foram utilizadas, a partir de cDNAs obtidos, técnicas de amplificação baseadas na Allele Specific Primer Extension (ASPE), visando detectar em teste multiplex os vírus mais relevantes envolvidos na etiologia da degenerescência e nos complexos do enrolamento das folhas e do lenho rugoso da videira. Os resultados obtidos mostraram que as técnicas agregam algumas vantagens, como a redução no tempo e relativa simplicidade de execução, eliminando completamente o uso de reagentes tóxicos, a exemplo do brometo de etídeo. O uso de multiplex facilita a amplificação de múltiplos alvos em uma única reação, reduzindo o tempo e o custo das análises.
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顔鴻儀 and Hung-yee Ngan. "Molecular diagnosis of penicilliosis marneffei." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31970035.
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Yau, Shu Ching. "Molecular diagnosis of neuromuscular disorders." Thesis, King's College London (University of London), 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.402033.
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Ngan, Hung-yee. "Molecular diagnosis of penicilliosis marneffei." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B23595978.
Full textBooks on the topic "Molecular Diagnosi"
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W, Semmler, and Schwaiger Markus, eds. Molecular imaging. Berlin: Springer, 2008.
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W, Semmler, and Schwaiger Markus, eds. Molecular imaging. Berlin: Springer, 2008.
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Jeffery, S. Molecular diagnosis. Oxford: BIOS Scientific, 1999.
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M, Kirkwood John, ed. Molecular diagnosis and treatment of melanoma. New York: M. Dekker, 1998.
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service), ScienceDirect (Online, ed. Molecular diagnostics. 2nd ed. Amsterdam: Elsevier/Academic Press, 2010.
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Molecular diagnostics of infectious diseases. Berlin: De Gruyter, 2010.
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1954-, Srivastava Sudhir, ed. Early detection of cancer: Molecular markers. Armonk, N.Y: Futura Pub. Co., 1994.
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Yousef, George M., and Serge Jothy. Molecular testing in cancer. New York: Springer, 2014.
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W, Semmler, and Schwaiger Markus, eds. Impact of molecular biology and new technical developments on diagnostic imaging. Berlin: Springer, 1997.
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Cotter, Finbarr. Molecular Diagnosis of Cancer. New Jersey: Humana Press, 1996. http://dx.doi.org/10.1385/0896033414.
Full textBook chapters on the topic "Molecular Diagnosi"
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García-Elorriaga, Guadalupe, and Guillermo del Rey-Pineda. "Molecular Diagnosis." In Practical and Laboratory Diagnosis of Tuberculosis, 35–53. Cham: Springer International Publishing, 2015. http://dx.doi.org/10.1007/978-3-319-20478-9_4.
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Hangay, George, Susan V. Gruner, F. W. Howard, John L. Capinera, Eugene J. Gerberg, Susan E. Halbert, John B. Heppner, et al. "Molecular Diagnosis." In Encyclopedia of Entomology, 2449–55. Dordrecht: Springer Netherlands, 2008. http://dx.doi.org/10.1007/978-1-4020-6359-6_4659.
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Weisman, Paul, Jian-Jun Wei, and Pei Hui. "Molecular Diagnosis." In Practical Gynecologic Pathology, 417–31. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-68608-6_16.
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Sloan, Philip, and Max Robinson. "Promising Biomarkers for Early Diagnosis and Prognosis Prediction." In Critical Issues in Head and Neck Oncology, 3–12. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-63234-2_1.
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AbstractSingle molecule biomarkers are used extensively in head and neck pathology for diagnosis and increasingly for prognosis. Companion markers for therapy such as PDL-1 and NTRK are now finding applications in head and neck cancer care. Immunohistochemistry is an attractive option because of its rapid turnaround time and convenience but molecular testing is often necessary for validation. This chapter will focus on some selected biomarkers being developed for translational purposes. Adoptive T cell therapies are being trialled for head and neck cancer and have limited efficacy currently. Identification of biomarkers as targets is an attractive option for development, and the use of molecular sequencing to identify individual neo-antigens is a promising way forward for precision medicine approaches including adoptive T cell therapies.
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Gronthoud, Firza Alexander. "Understanding Molecular Diagnosis." In Practical Clinical Microbiology and Infectious Diseases, 75–77. First edition. | Boca Raton : CRC Press, 2020.: CRC Press, 2020. http://dx.doi.org/10.1201/9781315194080-2-10.
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Liu, Chengcheng, Xiaoting Lou, Jianxin Lyu, Jian Wang, and Yufei Xu. "Prenatal Diagnosis and Preimplantation Genetic Diagnosis." In Clinical Molecular Diagnostics, 769–800. Singapore: Springer Singapore, 2021. http://dx.doi.org/10.1007/978-981-16-1037-0_43.
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Kardon, Nataline, and Lisa Edelmann. "Prenatal Diagnosis." In Molecular Genetic Pathology, 441–48. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-405-6_17.
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Huang, James, Sharie B. Parks, and Richard D. Press. "Diagnostic Molecular Pathology." In Essentials of Anatomic Pathology, 3–35. Totowa, NJ: Humana Press, 2006. http://dx.doi.org/10.1007/978-1-60327-173-8_1.
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Richards, C. Sue, and Patricia A. Ward. "Molecular Diagnostic Testing." In Principles of Molecular Medicine, 83–88. Totowa, NJ: Humana Press, 1998. http://dx.doi.org/10.1007/978-1-59259-726-0_8.
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Huang, James, and Richard D. Press. "Molecular Diagnostic Pathology." In Essentials of Anatomic Pathology, 489–513. New York, NY: Springer New York, 2010. http://dx.doi.org/10.1007/978-1-4419-6043-6_11.
Full textConference papers on the topic "Molecular Diagnosi"
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Litau, I. S., M. V. Alvarez Figueroa, A. A. Kazyulina, and L. V. Domotenko. "EVALUATION OF ANALYTICAL CHARACTERISTICS OF TB DIAGNOSTIC REAGENT KITS ON DOMESTIC CONTROL PANEL OF EXTERNAL QUALITY ASSESSMENT SAMPLES." In Molecular Diagnostics and Biosafety. Federal Budget Institute of Science 'Central Research Institute for Epidemiology', 2020. http://dx.doi.org/10.36233/978-5-9900432-9-9-216.
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The WHO tuberculosis eradication strategy includes early diagnosis of the disease through rapid diagnostic tests using molecular diagnostic techniques. For their correct use, it is necessary to improve the quality of laboratory services, including external quality assessment (EQA). In the course of the study on evaluation of analytical characteristics of TB diagnostic kits, 100% sensitivity and specificity are shown on the domestic control panel of EQA samples over a 5-year period. When determining the reproducibility of both sets of reagents, the CV did not exceed 15%.
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Sahu, Aditi, Ucalene Harris, Melissa Gill, Cecelia Lezcano, Allan Halpern, Anthony Rossi, Klaus Busam, et al. "PARP1 as a biomarker towards in vivo multimodal melanoma diagnosis." In Molecular-Guided Surgery: Molecules, Devices, and Applications VIII, edited by Summer L. Gibbs, Brian W. Pogue, and Sylvain Gioux. SPIE, 2022. http://dx.doi.org/10.1117/12.2610476.
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Chumachkova, Е. A., S. A. Portenko, E. S. Kazakova, O. V. Kedrova, S. A. Shcherbakova, T. L. Voznyuk, I. I. Krupinskaya, and R. N. Steshenko. "EPIDEMIOLOGICAL FEATURES OF THE COURSE OF COMMUNITY-ACQUIRED PNEUMONIA IN PATIENTS OF THE CITY OF SARATOV AND SARATOV OBLAST." In Molecular Diagnostics and Biosafety. Federal Budget Institute of Science 'Central Research Institute for Epidemiology', 2020. http://dx.doi.org/10.36233/978-5-9900432-9-9-186.
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The aim of the study was to identify the epidemiological features of the course of community-acquired pneumonia among patients in the city of Saratov and the Saratov region during the epidemic of a new coronavirus infection. The medical records of 129 patients of various ages with a confirmed diagnosis of community-acquired pneumonia, who were on inpatient and outpatient treatment, were analyzed. The study established the predominance of pneumonia caused by COVID-19, compared to pneumonia caused by other etiological agents, and the pattern of identification of mild, moderate and severe forms of the disease from the moment the first signs of the disease appear. The computed tomography of the chest organs was proved to be the most informative diagnostic method. The severity of the inflammatory process in the lungs was analyzed depending on the age of the patient, gender, and the presence of concomitant chronic diseases.
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Scholler, Jules, Olivier Thouvenin, Emilie Benoit a la Guillaume, and Claude Boccara. "One hundred percent successful automatic breast cancer diagnosis using static and dynamic FFOCT images (Conference Presentation)." In Molecular-Guided Surgery: Molecules, Devices, and Applications VI, edited by Summer L. Gibbs, Brian W. Pogue, and Sylvain Gioux. SPIE, 2020. http://dx.doi.org/10.1117/12.2544301.
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Leblond, Frédéric. "Label-free optical spectroscopy techniques for targeted, real time and informed in situ cancer diagnosis (Conference Presentation)." In Molecular-Guided Surgery: Molecules, Devices, and Applications IV, edited by Greg Biggs, Brian W. Pogue, and Sylvain Gioux. SPIE, 2018. http://dx.doi.org/10.1117/12.2287890.
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Albadawy, Ehab, Ashirbani Saha, Jun Zhang, Michael R. Harowicz, Maciej A. Mazurowski, and Zhe Zhu. "Breast cancer molecular subtype classification using deep features: preliminary results." In Computer-Aided Diagnosis, edited by Kensaku Mori and Nicholas Petrick. SPIE, 2018. http://dx.doi.org/10.1117/12.2295471.
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Tsourkas, Andrew, Jason Xu, and Gang Bao. "Hybridization Dynamics and Kinetics of Fret-Enhanced Molecular Beacons." In ASME 2001 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2001. http://dx.doi.org/10.1115/imece2001/bed-23163.
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Abstract Many human diseases start with a defect in the genome. Cancer, for example, is a genetic disease that arises from a single cell that behaves abnormally, dividing uncontrollably and leading, eventually, to the development of a tumor. A critical step in diagnosing and treating cancer is to detect cancer cells that result from the mutated genes. In spite of the extensive biomedical research efforts during the last few decades, it is still difficult to detect cancer at its early stages — when a cancer is diagnosed it is often too late to cure. A novel way of achieving early detection of cancer is to detect mRNA transcripts that arise from mutated genes in living cells [1]. We have developed a FRET-enhanced molecular beacons methodology which, combined with the state-of-the-art fluorescence imaging techniques, has the potential to detect cancer cells. FRET (Fluorescence resonance energy transfer) refers to the non-radiative transfer of energy from a donor molecule to an acceptor molecule through dipole-dipole coupling. As shown schematically in Figure 1, molecular beacons are dual labeled antisense oligonucleotides (ODNs) with a fluorophore (A or D) at one end and a quencher (Q) at the other; they are designed to form a hairpin structure in the absence of a complimentary target such that fluorescence of the fluorophore is quenched. Upon hybridization with the target mRNA, the molecular beacon opens up, leading to fluorescence [2,3].
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House, Broderick J., Jasmin M. Schaefer, Connor W. Barth, Scott C. Davis, and Summer L. Gibbs. "Diagnostic performance of receptor-specific surgical specimen staining correlate with receptor expression level." In Molecular-Guided Surgery: Molecules, Devices, and Applications V, edited by Brian W. Pogue and Sylvain Gioux. SPIE, 2019. http://dx.doi.org/10.1117/12.2510625.
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Giannelli, B. F. "MOLECULAR GENETICS OF HAEMOPHILIA." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643981.
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Haemophilia B, an X-linked recessive disease with an incidence of 1/30,000 newborn males, is due to defects in the gene for coagulation factor IX, which is on the long am of the X chromosome at band Xq27.1. This gene consists of approximately 34 Kb and contains 8 exons which specify a mRtfc of 2803 residues coding for a protein of 415 aa preceded by a prepro signal peptide of 46 aa. Coripanson of the functional domains of the factor IX protein with the exon structure of the gene supports the exon/protein domain hypothesis of gene evolution. The factor IX gene seems to be formed by a number of functionally and evolutionally independent modules. The signal peptide and the gla (γcarboxy-glutamic) region encoded in the first three exons are homologous to those of factor X, protein C and prothrombin. Thevfourth and fifth exons which code for the connecting peptide are homologous to one another and to the epidermal growth factor, a module that has been used in the construction of a great variety of proteins including different members of the coagulation and fibrinolytic pathways. The sixth exon encodes the activation peptide region, while the catalytic region of factor IX is coded by the seventh and eighth exon. This is at variance with other serine protease genes that have different exons for the segments containing the cardinal ami no-acids of the active centre (histidine, aspartic acid and serine).Natural selection acts against detrimental mutations of the factor IX gene and at each generation a proportion of haemophilia B genes is eliminated, as a significant number of patients does not reproduce. There appears to be no selective advantage to the heterozygote and therefore haemophilia B is maintained in the population by new mutations. Consequently, a significant proportion of patients should be born to non-carrier mothers, and unrelated patients should carry different gene defects, as recently verified by detailed analysis of individual haemophilia B genes.The defects of factor IX described so far comprise both point mutations and gene deletions. The latter affect either part or the whole of the gene and are often associated with the development of antibodies against therapeutically adninistered factor IX (the inhibitor complication). Since gene deletions may result in the complete absenceof factor IX synthesis or in the production of an extremely abnormal product, it has been suggested that mutationspreventing the synthesis of a factor IX gene product capable of inducing immune tolerance to normal factor IX is important in predisposing to the inhibitor complication.Among the point mutations described so far, those affecting the signal peptide are of particular interest. Substitutions of the arginine at positions -4 and -1 cause failure of propeptide cleavage. Thus they indicate that the propeptide consists of 18 aa an(lthat lts excision is necessary for factor IX function. It appears also that the propeptide contains a signal for γcarboxylation which has been conserved during the evolution of different γcarboxylated proteins.In spite of coagulant treatment, haemophilia B is a serious disease and one for which genetic counselling is required. Paramount for this is the detection of carriers and the diagnosis ofaffected male fetuses. DNA probes derived from the cloned factor IX gene have been used for this purpose. Carrier and first or second trimester prenatal diagnoses have been done using factors IX gene markers to follow the transmission of haemophilia B genes. Six sequence variations causing restriction fragment length polymorphisms (RFLP) in the factor IX gene have been detected and used as markers for such indirect diagnoses The efficiency of the above markers is reduced by linkage disequilibrium but, nevertheless, they offer definite carrier and nremtal diagnoses in 75-80% of the relatives of familial cases of haemophilia B.The indirect detection of gene defects is of modest help in the counselling of individuals from the families of isolated patients, but new methods for the direct detection of gene mutations promise better results in such families and also the attainment of % diagnostic success in relatives of familial cases.Finally the successful expression of recombinant factor IX genes in tissue culture and transgenic mammals raises hopes of therapeutic advances.
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Hu, Hui, and Manoochehr Koochesfahani. "Molecular Tagging Techniques for Micro-Flow and Micro-Scale Heat Transfer Studies." In ASME 2009 Fluids Engineering Division Summer Meeting. ASMEDC, 2009. http://dx.doi.org/10.1115/fedsm2009-78059.
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We report recent progresses made in development of novel molecule-based flow diagnostic techniques, named as Molecular Tagging techniques, to achieve simultaneous measurements of multiple important flow variables (such as flow velocity and temperature) for micro-flows and micro-scale heat transfer studies. Instead of using tiny particles, specially-designed phosphorescent molecules, which can be turned into long-lasting glowing molecules upon excitation by photons of appropriate wavelength, are used as tracers for both velocity and temperature measurements. A pulsed laser is used to “tag” the tracer molecules in the regions of interest, and the movements of the tagged molecules are imaged at two successive times within the photoluminescence lifetime of the tracer molecules. The measured Lagrangian displacement of the tagged molecules between the two image acquisitions provides the estimate of the fluid velocity vector. The simultaneous temperature measurement is achieved by taking advantage of the temperature dependence of phosphorescence lifetime, which is estimated from the intensity ratio of the tagged molecules in the two images. The implementation and application of the MTV&T technique are demonstrated by conducting simultaneous velocity and temperature measurements to qunatify the transient behavior of electroosmotic flow (EOF) inside a microchannel and to reveal the unsteady heat transfer, mass transfer and phase changing process inside micro-sized water droplets pertinent to wind turbine icing phenomena.
Reports on the topic "Molecular Diagnosi"
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Chen, Wen-Tien. Molecular Diagnosis for Breast Malignancy. Fort Belvoir, VA: Defense Technical Information Center, July 1998. http://dx.doi.org/10.21236/ada381191.
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Rinchik, E. M. Workshop on molecular methods for genetic diagnosis. Final technical report. Office of Scientific and Technical Information (OSTI), July 1997. http://dx.doi.org/10.2172/501564.
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Reinholz, Monica M., and Patrick C. Roche. Molecular Detection of Circulating Cancer Cells for Early Diagnosis of Breast Cancer. Fort Belvoir, VA: Defense Technical Information Center, October 2001. http://dx.doi.org/10.21236/ada400500.
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Lokshin, Anna. Integrated Development of Serum Molecular Markers for Early Diagnosis of Breast Cancer. Fort Belvoir, VA: Defense Technical Information Center, September 2006. http://dx.doi.org/10.21236/ada462774.
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Levisohn, Sharon, Maricarmen Garcia, David Yogev, and Stanley Kleven. Targeted Molecular Typing of Pathogenic Avian Mycoplasmas. United States Department of Agriculture, January 2006. http://dx.doi.org/10.32747/2006.7695853.bard.
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Intraspecies identification (DNA "fingerprinting") of pathogenic avian mycoplasmas is a powerful tool for epidemiological studies and monitoring strain identity. However the only widely method available for Mycoplasma gallisepticum (MG) and M. synoviae (MS)wasrandom amplified polymorphic DNA (RAPD). This project aimed to develop alternative and supplementary typing methods that will overcome the major constraints of RAPD, such as the need for isolation of the organism in pure culture and the lack of reproducibility intrinsic in the method. Our strategy focussed on recognition of molecular markers enabling identification of MG and MS vaccine strains and, by extension, pathogenic potential of field isolates. Our first aim was to develop PCR-based systems which will allow amplification of specific targeted genes directly from clinical material. For this purpose we evaluated the degree of intraspecies heterogeneity in genes encoding variable surface antigens uniquely found in MG all of which are putative pathogenicity factors. Phylogenic analysis of targeted sequences of selected genes (pvpA, gapA, mgc2, and lp) was employed to determine the relationship among MG strains.. This method, designated gene targeted sequencing (GTS), was successfully employed to identify strains and to establish epidemiologically-linked strain clusters. Diagnostic PCR tests were designed and validated for each of the target genes, allowing amplification of specific nucleotide sequences from clinical samples. An mgc2-PCR-RFLP test was designed for rapid differential diagnosis of MG vaccine strains in Israel. Addressing other project goals, we used transposon mutagenesis and in vivo and in vitro models for pathogenicity to correlated specific changes in target genes with biological properties that may impact the course of infection. An innovative method for specific detection and typing of MS strains was based on the hemagglutinin-encoding gene vlhA, uniquely found in this species. In parallel, we evaluated the application of amplified fragment length polymorphism (AFLP) in avian mycoplasmas. AFLP is a highly discriminatory method that scans the entire genome using infrequent restriction site PCR. As a first step the method was found to be highly correlated with other DNA typing methods for MG species and strain differentiation. The method is highly reproducible and relatively rapid, although it is necessary to isolate the strain to be tested. Both AFLP and GTS are readily to amenable to computer-assisted analysis of similarity and construction of a data-base resource. The availability of improved and diverse tools will help realize the full potential of molecular typing of avian mycoplasmas as an integral and essential part of mycoplasma control programs.
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Svetlov, Stanislav, Ronald Hayes, and Olena Glushakova. Molecular Signatures and Diagnostic Biomarkers of Cumulative Blast-Graded Mild TBI. Fort Belvoir, VA: Defense Technical Information Center, December 2014. http://dx.doi.org/10.21236/ada612707.
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Svetlov, Stanislav. Molecular Signatures and Diagnostic Biomarkers of Cumulative, Blast-Graded Mild TBI. Fort Belvoir, VA: Defense Technical Information Center, October 2012. http://dx.doi.org/10.21236/ada582352.
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Osburn, Bennie, Marius Ianconescu, Geoffrey Akita, and Rozalia Kaufman. Rapid, Sensitive Bluetongue Virus Serogroup and Serotype Detection Using Polymerase Chain Reaction. United States Department of Agriculture, September 1995. http://dx.doi.org/10.32747/1995.7612836.bard.
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The objectives of this proposal were to enhance animal health by 1) development of a BTV serogroup diagnostic assay using polymerase chain reaction (PCR) and 2) development of a BTV serotype specific diagnostic PCR assay. A PCR assay for diagnosis of bluetongue virus (BTV) serogroup from clinical samples meeting the criteria of objective 1 was developed. This PCR assay is more sensitive than virus isolation and has been adopted by both the U.S. and Israeli collaborating laboratories of this project, as well as at least one other U.S. laboratory for routine diagnosis of BTV infection in ruminants. The basic BTV PCR protocol has also become an essential tool in BTV molecular research in both collaborating laboratories. During development of the BTV serotype specific PCR we had the opportunity to investigate a nationwide outbreak of abortions and fatal disease in dogs in the U.S. purportedly due to BTV infection via a BTV contaminated canine vaccine. The BTV serogroup PCR was integral in confirming BTV in tissues from affected dogs and in lots of the suspect vaccine. This led to the first published report of BTV infection in dogs. We discovered that BTV can produce silent persistent infection in canine cell culture. This indicated a need for more stringent screening of biologics for occult BTV infection. A novel mixed cell culture method was developed to identify occult BTV and other occult viral infection cell cultures. Serotype specific primers for PCR detection of all U.S. BTV serotypes and two Israel serotypes (BTV-2 and 10) have been evaluated and are available. A subsequent collaboration would logically include sequencing of the L2 genes of Israel BTV-4, 6 and 16, allowing incorporation of these Israel BTV serotypes into a multiplex PCR assay.
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Splitter, Gary, and Menachem Banai. Microarray Analysis of Brucella melitensis Pathogenesis. United States Department of Agriculture, 2006. http://dx.doi.org/10.32747/2006.7709884.bard.
Full textAbstract:
Original Objectives 1. To determine the Brucella genes that lead to chronic macrophage infection. 2. To identify Brucella genes that contribute to infection. 3. To confirm the importance of Brucella genes in macrophages and placental cells by mutational analysis. Background Brucella spp. is a Gram-negative facultative intracellular bacterium that infects ruminants causing abortion or birth of severely debilitated animals. Brucellosis continues in Israel, caused by B. melitensis despite an intensive eradication campaign. Problems with the Rev1 vaccine emphasize the need for a greater understanding of Brucella pathogenesis that could improve vaccine designs. Virulent Brucella has developed a successful strategy for survival in its host and transmission to other hosts. To invade the host, virulent Brucella establishes an intracellular niche within macrophages avoiding macrophage killing, ensuring its long-term survival. Then, to exit the host, Brucella uses placenta where it replicates to high numbers resulting in abortion. Also, Brucella traffics to the mammary gland where it is secreted in milk. Missing from our understanding of brucellosis is the surprisingly lillie basic information detailing the mechanisms that permit bacterial persistence in infected macrophages (chronic infection) and dissemination to other animals from infected placental cells and milk (acute infection). Microarray analysis is a powerful approach to determine global gene expression in bacteria. The close genomic similarities of Brucella species and our recent comparative genomic studies of Brucella species using our B. melitensis microarray, suqqests that the data obtained from studying B. melitensis 16M would enable understanding the pathogenicity of other Brucella organisms, particularly the diverse B. melitensis variants that confound Brucella eradication in Israel. Conclusions Results from our BARD studies have identified previously unknown mechanisms of Brucella melitensis pathogenesis- i.e., response to blue light, quorum sensing, second messenger signaling by cyclic di-GMP, the importance of genomic island 2 for lipopolysaccharide in the outer bacterial membrane, and the role of a TIR domain containing protein that mimics a host intracellular signaling molecule. Each one of these pathogenic mechanisms offers major steps in our understanding of Brucella pathogenesis. Strikingly, our molecular results have correlated well to the pathognomonic profile of the disease. We have shown that infected cattle do not elicit antibodies to the organisms at the onset of infection, in correlation to the stealth pathogenesis shown by a molecular approach. Moreover, our field studies have shown that Brucella exploit this time frame to transmit in nature by synchronizing their life cycle to the gestation cycle of their host succumbing to abortion in the last trimester of pregnancy that spreads massive numbers of organisms in the environment. Knowing the bacterial mechanisms that contribute to the virulence of Brucella in its host has initiated the agricultural opportunities for developing new vaccines and diagnostic assays as well as improving control and eradication campaigns based on herd management and linking diagnosis to the pregnancy status of the animals. Scientific and Agricultural Implications Our BARD funded studies have revealed important Brucella virulence mechanisms of pathogenesis. Our publication in Science has identified a highly novel concept where Brucella utilizes blue light to increase its virulence similar to some plant bacterial pathogens. Further, our studies have revealed bacterial second messengers that regulate virulence, quorum sensing mechanisms permitting bacteria to evaluate their environment, and a genomic island that controls synthesis of its lipopolysaccharide surface. Discussions are ongoing with a vaccine company for application of this genomic island knowledge in a Brucella vaccine by the U.S. lab. Also, our new technology of bioengineering bioluminescent Brucella has resulted in a spin-off application for diagnosis of Brucella infected animals by the Israeli lab by prioritizing bacterial diagnosis over serological diagnosis.
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Dawson, William O., Moshe Bar-Joseph, Charles L. Niblett, Ron Gafny, Richard F. Lee, and Munir Mawassi. Citrus Tristeza Virus: Molecular Approaches to Cross Protection. United States Department of Agriculture, January 1994. http://dx.doi.org/10.32747/1994.7570551.bard.
Full textAbstract:
Citrus tristeza virus (CTV) has the largest genomes among RNA viruses of plants. The 19,296-nt CTV genome codes for eleven open reading frames (ORFs) and can produce at least 19 protein products ranging in size from 6 to 401 kDa. The complex biology of CTV results in an unusual composition of CTV-specific RNAs in infected plants which includes multiple defective RNAs and mixed infections. The complex structure of CTV populations poses special problems for diagnosis, strain differentiation, and studies of pathogenesis. A manipulatable genetic system with the full-length cDNA copy of the CTV genome has been created which allows direct studies of various aspects of the CTV biology and pathology. This genetic system is being used to identify determinants of the decline and stem-pitting disease syndromes, as well as determinants responsible for aphid transmission.