A MALDI-TOF alkalmazása fertőző betegségek diagnosztizálásában
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A MALDI-TOF alkalmazása fertőző betegségek diagnosztizálásában
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A kórokozó fertőzéses betegségek genotípusos vizsgálati módszerei
A kórokozó fertőzéses betegségek genotípusos vizsgálati módszerei
In vitro models of hepatotoxicity
This document describes various in vitro models and methods that can be used to study hepatotoxicity, including hepatocyte cell cultures, assays to measure cell viability and metabolic activity (trypan blue dye exclusion test, MTT assay), staining to visualize lipid accumulation (Oil Red O), and techniques to examine gene and protein expression changes (RT-PCR, western blotting). Specifically, it discusses using these methods to establish models of non-alcoholic fatty liver disease (NAFLD) by treating hepatocyte cultures with fatty acids like palmitic and oleic acid, and models of drug-induced hepatotoxicity by treating with acetaminophen or amiodarone. Key readouts include lipid accumulation, apoptosis levels
Etiology of liver diseases
This document summarizes various liver diseases and their etiologies. It discusses alcoholic liver disease, drug-induced liver injury, viral hepatitis infections from hepatitis B, C, and D viruses, autoimmune disorders like autoimmune hepatitis and primary biliary cirrhosis, genetic disorders, non-alcoholic fatty liver disease, cirrhosis, and hepatocellular carcinoma. The liver's important functions are outlined. Causes, pathogenesis, clinical features, diagnosis, and treatment approaches are described for each disease.
An introduction to experimental epidemiology
This document provides an overview of experimental epidemiology methods. It discusses the key features and types of experimental epidemiology studies, including controlled field trials and community trials. Controlled field trials involve dividing healthy subjects into an exposed group that receives an active substance (like a vaccine) and an unexposed control group that receives a placebo. Community trials involve entire exposed and unexposed communities. Randomized controlled trials, which assign individual subjects randomly to intervention or control groups, are described as the most common experimental method but are covered in more depth separately. Overall, the document outlines the design and purpose of various experimental epidemiology study types.
Genotyping methods of nosocomial infections pathogen
Nosocomial infections afflict around 2 million patients in the US each year, resulting in around 88,000 deaths and $4.5 billion in excess healthcare costs. Understanding the distribution and relatedness of pathogens that cause these infections is important for designing effective control methods. Historically, phenotypic characterization was used, but increasingly molecular or genotyping techniques are being used, including pulsed-field gel electrophoresis, multilocus sequence typing, and polymerase chain reaction-based methods. Studies have shown that integrating molecular typing into infection control programs can significantly reduce infection rates and healthcare costs.
Use of MALDI-TOF in the diagnosis of infectious diseases
MALDI-TOF MS has revolutionized clinical microbiology by drastically improving the time needed to identify bacterial cultures from over 24 hours to just a few minutes. Whereas the entire process from sampling to results previously took 2-3 days or more, new methods like MALDI-TOF MS and molecular technology have reduced this to just a few hours or one day. MALDI-TOF MS is a powerful, cost-effective, and easy to implement technique that provides rapid and reliable identification of bacteria and yeast from clinical samples at the genus and species level through analysis of their protein mass spectral signatures.
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A kórokozó fertőzéses betegségek genotípusos vizsgálati módszerei
A kórokozó fertőzéses betegségek genotípusos vizsgálati módszerei
In vitro models of hepatotoxicity
This document describes various in vitro models and methods that can be used to study hepatotoxicity, including hepatocyte cell cultures, assays to measure cell viability and metabolic activity (trypan blue dye exclusion test, MTT assay), staining to visualize lipid accumulation (Oil Red O), and techniques to examine gene and protein expression changes (RT-PCR, western blotting). Specifically, it discusses using these methods to establish models of non-alcoholic fatty liver disease (NAFLD) by treating hepatocyte cultures with fatty acids like palmitic and oleic acid, and models of drug-induced hepatotoxicity by treating with acetaminophen or amiodarone. Key readouts include lipid accumulation, apoptosis levels
Etiology of liver diseases
This document summarizes various liver diseases and their etiologies. It discusses alcoholic liver disease, drug-induced liver injury, viral hepatitis infections from hepatitis B, C, and D viruses, autoimmune disorders like autoimmune hepatitis and primary biliary cirrhosis, genetic disorders, non-alcoholic fatty liver disease, cirrhosis, and hepatocellular carcinoma. The liver's important functions are outlined. Causes, pathogenesis, clinical features, diagnosis, and treatment approaches are described for each disease.
An introduction to experimental epidemiology
This document provides an overview of experimental epidemiology methods. It discusses the key features and types of experimental epidemiology studies, including controlled field trials and community trials. Controlled field trials involve dividing healthy subjects into an exposed group that receives an active substance (like a vaccine) and an unexposed control group that receives a placebo. Community trials involve entire exposed and unexposed communities. Randomized controlled trials, which assign individual subjects randomly to intervention or control groups, are described as the most common experimental method but are covered in more depth separately. Overall, the document outlines the design and purpose of various experimental epidemiology study types.
Genotyping methods of nosocomial infections pathogen
Nosocomial infections afflict around 2 million patients in the US each year, resulting in around 88,000 deaths and $4.5 billion in excess healthcare costs. Understanding the distribution and relatedness of pathogens that cause these infections is important for designing effective control methods. Historically, phenotypic characterization was used, but increasingly molecular or genotyping techniques are being used, including pulsed-field gel electrophoresis, multilocus sequence typing, and polymerase chain reaction-based methods. Studies have shown that integrating molecular typing into infection control programs can significantly reduce infection rates and healthcare costs.
Use of MALDI-TOF in the diagnosis of infectious diseases
MALDI-TOF MS has revolutionized clinical microbiology by drastically improving the time needed to identify bacterial cultures from over 24 hours to just a few minutes. Whereas the entire process from sampling to results previously took 2-3 days or more, new methods like MALDI-TOF MS and molecular technology have reduced this to just a few hours or one day. MALDI-TOF MS is a powerful, cost-effective, and easy to implement technique that provides rapid and reliable identification of bacteria and yeast from clinical samples at the genus and species level through analysis of their protein mass spectral signatures.
Molecular microbiology methods
1. Molecular microbiology methods like PCR and hybridization have revolutionized clinical diagnostics by enabling fast and direct detection of pathogens from clinical samples.
2. PCR in particular has become a mainstay technique, allowing amplification of specific DNA sequences from small amounts of input DNA. Variations like real-time PCR, multiplex PCR, and broad-range PCR further expanded diagnostic capabilities.
3. Emerging technologies like DNA microarrays promise even greater multiplexing, with the ability to simultaneously genotype large genomic regions or measure expression of many genes, positioning them as promising future molecular diagnostic tools.
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2. Explaining the preparation of Krebs-Hanseleit solution and common drugs used.
3. Outlining typical experiment protocols, including stabilizing tissues, pre-contraction testing, and assessing endothelial function.
4. Noting that each experiment begins by preparing Krebs-Hanseleit solution and activating the system before surgery and setting rings in wells.
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1) Early diagnosis of cancers like lung cancer and breast cancer through predictive modeling of patient data to detect cancers at earlier stages when survival rates are higher.
2) Predicting patient responses to drug therapies for cancers like breast cancer by combining different types of molecular profiling data using techniques like support vector machines and random forests.
3) Using imaging data and temporal analysis of metrics like medication purchases to better understand and predict chronic diseases like diabetes and associated health complications.
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2. B cells undergo gene rearrangement and development in the bone marrow before migrating to peripheral lymphoid organs. T cells develop through similar processes in the thymus.
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1. Molecular microbiology methods like PCR and hybridization have revolutionized clinical diagnostics by enabling fast and direct detection of pathogens from clinical samples.
2. PCR in particular has become a mainstay technique, allowing amplification of specific DNA sequences from small amounts of input DNA. Variations like real-time PCR, multiplex PCR, and broad-range PCR further expanded diagnostic capabilities.
3. Emerging technologies like DNA microarrays promise even greater multiplexing, with the ability to simultaneously genotype large genomic regions or measure expression of many genes, positioning them as promising future molecular diagnostic tools.
Isolated vascular rings
This document provides information about setting up and conducting experiments with isolated organs and tissue rings, including:
1. Describing the mechanical setup for a four-channel system bath for isolated organs.
2. Explaining the preparation of Krebs-Hanseleit solution and common drugs used.
3. Outlining typical experiment protocols, including stabilizing tissues, pre-contraction testing, and assessing endothelial function.
4. Noting that each experiment begins by preparing Krebs-Hanseleit solution and activating the system before surgery and setting rings in wells.
Isolated blood vessels
This document describes the components, work principles, and experimental protocols for using a pressure myograph system to study isolated blood vessels. The system allows measuring vessel diameter in response to drugs and stimuli while maintaining constant temperature. Experiments involve isolating small arteries from rats and attaching them to glass micropipettes in a chamber filled with physiological salt solution. Vessel diameter is recorded under varying pressures and drug exposures to study endothelial function and vasoactive mechanisms. Statistical analysis of diameter changes under different conditions uses repeated measures ANOVA to compare responses between experimental groups.
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This document describes the laser Doppler flowmetry (LDF) method for measuring blood flow in the microcirculation of skin. Specifically, it discusses post-occlusive reactive hyperemia (PORH) testing using LDF to assess microvascular reactivity by inducing a brief occlusion of blood vessels. It also covers iontophoresis of acetylcholine and sodium nitroprusside combined with LDF to evaluate endothelium-dependent and independent vasodilation respectively. Standardization of methods like occlusion duration and probe placement is important for reproducibility. LDF provides a general index of microvascular function rather than direct flow measurements.
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
This document provides an overview of histological staining techniques. It discusses how histological preparations are stained using interactions between dyes, solvents, and tissue components. Different staining methods result in different colors that highlight various structures. A classic example is hematoxylin and eosin staining, where hematoxylin stains acidic components blue and eosin stains basic components pink. Specialized staining techniques also exist, such as immunohistochemistry. Proper staining selection depends on the tissue and research goals. Histological preparations are then analyzed under a microscope to study cell and tissue morphology.
Notes for Fixation of tissues and organs for educational and scientific purposes
Fixation of tissues and organs is done to preserve them for scientific and educational purposes. Various chemical fixatives are used including formaldehyde, alcohols, and acids. Formaldehyde cross-links proteins to harden the tissue while maintaining the original structure. Several fixation protocols are used for different purposes, balancing preservation of color and long-term durability. Key steps include diffusion or injection of fixatives, followed by storage in preservative solutions. Proper fixation and storage are necessary to prevent degradation over time.
Notes for
The document summarizes the process of preparing tissue samples for histological analysis, including fixation, dehydration, infiltration/embedding, sectioning, staining, and examination. Key steps involve fixing tissues to prevent degradation, dehydrating using increasing alcohol concentrations, infiltrating with paraffin wax or resin for structural support during sectioning, precisely cutting thin sections, mounting them to glass slides, staining, and examining under a microscope. The quality of prepared samples depends on carefully following each step of the preparation process.
Notes for The principle and performance of capillary electrophoresis
This document provides an overview of capillary electrophoresis (CE). It begins by introducing CE and its advantages over other separation techniques. It then describes the basic theory behind CE, including electrophoretic mobility, electroosmotic flow, and how samples migrate through the capillary when an electric field is applied. The document details the key components of a CE instrument and various CE separation techniques such as capillary zone electrophoresis, micellar electrokinetic chromatography, and capillary isoelectric focusing. It focuses on the principles and applications of CE.
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Big datasets
This document discusses systems biology and its goals of understanding how biological molecules interact and systems function as a whole. It covers:
1) Systems biology uses large datasets from "omics" experiments and computational models to understand complex biological interactions beyond individual molecules.
2) Pioneering work used microarrays to measure thousands of genes in serum-stimulated cells, finding over 500 changed in proliferation.
3) The field aims to discover emergent system properties and functions not evident from separate parts, like switches that change cell behavior.
Systems biology for Medicine' is 'Experimental methods and the big datasets
This document discusses experimental methods used in systems biology to generate large datasets, including microarrays, sequencing-based methods, mass spectrometry, and liquid chromatography. It explains that systems biology studies must be quantitative and enable computational modeling. Key methods covered are microarrays, RNA-seq, ChIP-seq, whole-genome sequencing, whole-exome sequencing, proteomics using mass spectrometry, and combining liquid chromatography with mass spectrometry for lipidomics, metabolomics and glycomics. Sources of variation are also discussed for genomic and proteomic studies.
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Use cases
The document discusses several use cases for applying data mining and machine learning techniques in healthcare and biomedical research. Three examples are:
1) Early diagnosis of cancers like lung cancer and breast cancer through predictive modeling of patient data to detect cancers at earlier stages when survival rates are higher.
2) Predicting patient responses to drug therapies for cancers like breast cancer by combining different types of molecular profiling data using techniques like support vector machines and random forests.
3) Using imaging data and temporal analysis of metrics like medication purchases to better understand and predict chronic diseases like diabetes and associated health complications.
Basic course for computer based methods
The document discusses various data mining methods. It describes data mining as seeking patterns within large databases. Common data mining methods mentioned include clustering, regression, rule extraction, and data visualization. Machine learning algorithms often used for health data include logistic regression, support vector machines, decision trees, and neural networks. The document also discusses newer techniques like graph-based data mining, topological data mining, and data visualization for exploring complex data.
Medicine as data science
This document discusses biomedical informatics and the increasing role of data in medicine. It notes that medicine is becoming a more data-intensive field due to growing sources of electronic health data. Biomedical data is often large in volume, diverse, complex, weakly structured, noisy, and inconsistent. Extracting knowledge from this "big data" through techniques like data mining, machine learning, and integrating human-computer interaction can provide insights to improve healthcare outcomes. Key applications include personalized and predictive medicine through patient stratification and risk analysis. However, overcoming obstacles like heterogeneous and non-standardized data is challenging.
Basic Immunology 21 26
This document discusses hypersensitivity reactions and autoimmune diseases. It describes the four types of hypersensitivity reactions according to the Gell and Coombs classification: Type I (immediate), Type II (cytotoxic), Type III (immune complex-mediated), and Type IV (delayed type hypersensitivity). It provides details on the mechanisms and examples of each type. The document then discusses immunological tolerance, including central and peripheral tolerance. It explains how a breakdown in tolerance can lead to autoimmune diseases and provides examples like Graves' disease, myasthenia gravis, hemolytic anemia, and systemic lupus erythematosus.
Basic Immunology 11-20
The document discusses lymphocyte development and antigen receptor gene rearrangement. It covers the following key points:
1. Lymphocyte development involves commitment to the B or T cell lineage, proliferation of progenitors, rearrangement of antigen receptor genes, selection checkpoints, and differentiation into distinct subpopulations.
2. B cells undergo gene rearrangement and development in the bone marrow before migrating to peripheral lymphoid organs. T cells develop through similar processes in the thymus.
3. During development, gene rearrangement generates diversity in antigen receptor genes, and selection checkpoints ensure that only lymphocytes with functional receptors will mature and enter the peripheral immune system.
Basic immunology 1 10
This document provides an overview of basic immunology concepts. It begins with definitions of key immunology terms like immunity, immunology, antigen, and discusses the historical figures Edward Jenner and Louis Pasteur who were pioneers in vaccination. It then discusses the components of the immune system including organs like the bone marrow, thymus, lymph nodes, and spleen. It provides information on cells of the immune system like antigen presenting cells, T and B lymphocytes, and effector cells. It also discusses molecular components of antigen recognition including antibodies, T cell receptors, B cell receptors, and the major histocompatibility complex.
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A MALDI-TOF alkalmazása fertőző betegségek diagnosztizálásában
- 1. Improved Medical Education and Basic sciences for Better Medical practicing ImproveMEd A MALDI-TOF alkalmazása fertőző betegségek diagnosztizálásában Domagoj Drenjancevic
- 2. A mikroorganizmus azonosítása • Fenotípusos tulajdonságok kimutatása • Gramfolt, kolónia • morfológia, mikroszkópos vizsgálat, szelektív táptalaj növekedési különbsége és különböző biokémiai vizsgálatok • kézi vagy automatizált módszerek • molekuláris diagnosztikai módszerek • 16S riboszomális RNS szekvenálás • a kiválasztott gének valós idejű PCR-detektálása • összetett és drága • MALDI-TOF MS Bizzini A, Greub G. Clin Microhiol Zaraziti 2010; 16: 1614-1619
- 3. MALDI-TOF MS: Mátrix-segítő lézer deszorpciós ionizációs idő a repülés tömegspektrometriáján - előnyök • megvalósítása a klinikai mikrobiológiai laboratóriumban - eASE használata • A MALDI-TOF MS bevezetése a baktériumok azonosításának alapja a rutin mintákból: • > 90% - hatékony • Megbízható eljárás a baktériumok klinikai mintákból történő azonosítására • költséghatékonyság: • az ár ¼-e a hagyományos módszerekhez képest • (2,44 euró MALDITOF MS-vel vs 4,60–13,85 € automatikus azonosítóval • Gyorsítási rendszer • 6 perc, • hagyományos technikák - ugyanaz az azonosítás 5-48 óra múlva • Helyi antibiotikum-rezisztencia adatokkal együtt használható • Hatékonyan optimalizálja a korai empirikus antibiotikum-kezelést
- 5. Clark AE i sur. Clin, Microhiol, Rev 2013; 26: 547-603
- 7. A MALDI TOF MS teljesítménye: ASOBIC típusú vizsgálat • Elméletileg nincs azonosító képesség korlátozás: ez szinte 100% -os helyes azonosítást eredményezett • Neisseria • clostridiumokhoz • mycobacteria • Nonfermenters • szalmonella • viridan streptococcus • Helicobacter pylori • élesztő (96% 250 klinikai Candida izolátum) • fonalas gombák és dermatofiták - Javítani kell az adatbázis-fejlesztéseket és a szükséges analitikai protokollokat
- 8. A MALDI TOF MS teljesítménye: A kórokozó közvetlen azonosítása a mintákban • a kórokozó közvetlen azonosítása a Sam mintában, tenyésztési lépés nélkül 66-90% -a helyes azonosítás
- 9. A MALDI TOF MS teljesítménye: antibiotikum érzékenységvizsgálat - folyamatában • A tömegspektrometria képessége megkülönbözteti a meticillin- érzékeny S. aureust a meticillin-rezisztens S. aureus-tól • a molekula különböző bakteriális antimikrobiális célzásai (mint például a b-laktamáz-metilázok és a pumpa-generáció) • 100% -os és 90,6% -os specificitású Pantone-Wavy leukocidin érzékenységének kimutatása
- 10. MALDI-RE (SEQUENOM) és elektroionizációs tömegspektrometria (ESI-MS) • alternatívája a tipizálási technológiának • a PCR amplifikációs eredményeinek elemzése Emonet S pri al, Clin Microhiol Zaraziti2010; 16: 1604-1613
- 11. MALDI - TOF Laboratóriumi integraciója E x P E C T A T ja O N S
- 12. Megállapítások • Sebesség, könnyű használhatóság és alacsony mintavételi ár • a klinikai mikrobiális diagnosztika forradalma