No description available.

There is growing appreciation of the potential for high intensity interval training (HIT) to rapidly stimulate metabolic adaptations that resemble traditional endurance training, despite a low total exercise volume. However, much of the work has been conducted on young active individuals and the results may not be applicable to older, less active populations. In addition, many studies have employed "all out", variable-load exercise interventions (e.g., repeated Wingate Tests) that may not be safe, practical or well tolerated by certain individuals. We determined the effect of a short program of low-volume, submaximal, constant-load HIT on skeletal muscle oxidative capacity and insulin sensitivity in sedentary middle-aged individuals who may be at higher risk for inactivity-related chronic diseases. Sedentary but otherwise healthy men (n=3) and women (n=4) with a mean (±SE) age, body mass index and peak oxygen uptake (VO_2peak) of 45±2 yr, 27±2 kg-m^2 and 30±1 ml·kg^-1·min^-1 were recruited. Subjects performed 6 training sessions over 2 wk, each consisting of 10 x 1 min cycling at 60% of peak power elicited during a ramp VO_2peak test (<90% of heart rate reserve) with 1 min recovery between intervals. Needle biopsy samples (v. lateralis) were obtained before training and <72 h after the final training session. Muscle oxidative capacity, as reflected by the maximal activity and protein content of citrate synthase, increased by ~20% after training, which is similar to changes previously reported after 2 wk of Wingate-based HIT in young active subjects. Insulin sensitivity, based on fasting glucose and insulin, improved by ~35% after training. These data support the notion that low-volume HIT may be a practical, time- efficient strategy to induce metabolic adaptations that reduce the risk for inactivity-related disorders in previously sedentary adults. / Thesis / Master of Science (MSc)

This dissertation describes the elucidation and implementation of derivatisation in the quantification of biologically active vitamin D metabolites in limited volume serum and dry blood spot samples (DBS) using the liquid chromatography tandem-mass spectrometry (LC-MS/MS) analytical technique. This manuscript describes in detail the development and validation of an analytical methodology, highlighting the role derivatisation and mass spectrometry plays in the structural characterisation and quantification of vitamin D metabolites. The first chapter reviews comprehensively, the history of vitamin D biosynthesis discovery as an anti-rickets agent, the biochemistry of vitamin D, its metabolic pathway, functions in the different biological systems and the consequences of its deficiency in the body. The second chapter reviews the current methods and techniques utilised for the detection and characterization of vitamin D metabolites, with specific emphasis based on the contribution made by derivatisation and mass spectrometry. A brief introduction to derivatisation is provided, with specific focus on PTAD and Amplifex diene reagents (Cooksontype reagents) used in this study. The importance of sensitivity and selectivity of targeted analytes is described first in detail for underivatised analytes, followed by PTAD and Amplifex derivatised samples. Chapter 2 also describes the importance of vitamin D quantification using liquid chromatography, the strengths and limitations of LC-MS/MS when used in isolation and after derivatisation. Also discussed, is how combining these techniques can overcome inherent limitations in LCMS/MS and enhance analytical performance. In Chapter 3 the materials and methods used and the study design is laid out, describing a brief introduction of the routinely used clinical diagnostics assay enzyme-linked immunosorbent assay (ELISA) as a reference method and is compared to an LC-MS/MS assay, to ascertain discrepancies and agreement between both methodologies from the same volunteer samples. Chapters 3 and 4 describes the comprehensive development, optimisation and validation of the highly sensitive PTAD derivatives LC-MS/MS assay for the quantification of active vitamin D metabolites, as well as the development of method using Amplifex diene derivatisation. Also discussed, is sample preparation optimisation of DBS and Mitra micro-samples. A holistic approach was taken to the development of the methodologies to provide data from which the required analytical information can be obtained for method evaluation and statistical analysis. The validated PTAD derivatives method is applied to the quantification of vitamin D metabolites in limited volume (100 μL) clinical human serum samples from 30 volunteers compared to results obtained using the clinical diagnostics ELISA technique. In Chapter 4 data analysis is described and the results are further discussed and a conclusion made based on the findings from the study. This study envisaged that combination of limited sample volume and DBS, derivatisation and LCMS/ MS is a powerful tool in vitamin D metabolite analysis and provided evidence of a positive increase in sensitivity and selectivity between derivatised compared to underivatised samples. A 10-fold increase in signal-to-noise-ratio (S/N) was observed when comparing PTAD derivatised, and Amplifex diene derivatised versus underivatised samples. Chapter 5 presents suggested future directions and considerations in the areas of vitamin D metabolite derivatisation and DBS sampling technique analysis using LC-MS/MS research based on the results presented in this dissertation. / Dissertation (MSc)--University of Pretoria, 2017. / Pharmacology / MSc / Unrestricted

Syftet med detta arbete är att fördjupa min kunskap om hur datorn kan användas som musikaliskt instrument. Datorer kan reproducera och simulera akustiska instrument men likväl kan den även skapa elektroakustiska ljud genom synthesizer och samplers. Inom det moderna musikskapandet är datorn ett ytterst viktigt instrument där den bland annat fungerar som dokumentationsmaskin, det vill säga vår tids bandspelare. Andra användningsområden av datorn i dagens musiklandskap är i ”live-framträdanden”. Med detta arbete vill jag praktisera och fördjupa mina kunskaper om hur datorn kan användas som instrument i mitt eget musicerande. Genom dokumentation i form av loggbok, video och ljudinspelningar kommer jag att analysera min process med avstamp inom det sociokulturella perspektivet. / The purpose of this work is to deepen my knowledge of how the computer can be used as a musical instrument. Computers can reproduce and simulate acoustic instruments but still they can also create electroacoustic sounds through synthesizers and samplers. In modern music creation, the computer is an extremely important instrument where, among other things, it serves as a tape-machine, the tape recorder of our time. Other uses of the computer in today's music landscape are in "live performances".   With this work I want to practice and deepen my knowledge of how the computer can be used as an instrument in my own music making. Through documentation in the form of logbook, video and audio recordings, I will analyze my process with an starting point in the socio-cultural perspective.

No description available.

Thesis (MMed)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Background: Sputum is a non-invasive, economic investigation whereby bronchogenic carcinoma can be identified. Manual cytological screening is labour intensive, time-consuming and requires a continuous high level of alertness. Automation has recently been successfully introduced in gynaecological cytology. Since sputum samples are similar to cervical smears, the question arises as to whether they are also suitable for automated screening. Objective: This study presented with various objectives: 1) To test automated sputum screening using the BD FocalPoint™ Slide Profiler (FP) and compare with manual sputum screening. 2) To determine the sensitivity and specificity of sputum in identification of bronchogenic carcinoma. 3) To ascertain if any clinical, radiological or bronchoscopy findings would be predictors for bronchogenic carcinoma. 4) To determine the significance of adequacy. Method: Sputum samples were collected prospectively from patients attending the Division of Pulmonology at Tygerberg hospital for a transbronchial fine needle aspiration biopsy (TBNA) or a transthoracic fine needle aspiration biopsy (TTNA) for the period from 2010 to 2012. A pre-bronchoscopy sputum was collected and submitted for processing. Stained slides were put through the FP for automated screening. After slides were qualified, sputum slides were put back in the routine screening pool. Correlation was done using the TBNA/TTNA result as the standard to evaluate the sputum results. Results: 108 sputum samples were included in this study. Of the 84.3% malignant (n=91) and 15.7% benign (n=17) cases confirmed with a diagnostic procedure, sputum cytology had a sensitivity of 38.5% (35/91 malignant cases), and a specificity of 100% (17/17 benign cases). Automated screening had a better sensitivity of 94.3% (33/35 positive sputum cases), while manual screening showed a sensitivity of 74.3% (26/35 positive sputum cases) when compared to the final sputum result. Individual parameters with a significant association with positive sputum included the presence of an endobronchial tumour, partial airway obstruction / stenosis, round mass, spiculated mass (negative association), loss of weight (negative association) and squamous cell carcinoma as the histological subtype. Adequacy was not as significant as hypothesised since 85.3% of true positive sputum, but also 65.5% of false negative sputum, had large numbers of alveolar macrophages present. Conclusion: Sputum cytology remains an important part of the screening programme for bronchogenic carcinoma in the public health sector of South Africa. Results confirm that sputum cytology is very specific, and automated screening improves sensitivity. Automated screening proved to be more time efficient, resulting in 83.1% reduction (p<0.0001) in the screening time spent per case by a cytotechnologist. Results confirm that the quantity of alveolar macrophages is not directly proprtional to pathology representation. Positive sputum results did however improve with sputum adequacy, but had no significant association. Recommendations from this study include adopting automated sputum screening. / AFRIKAANSE OPSOMMING: Agtergrond: Die verkryging van ‘n sputummonster is ‘n nie-indringende, ekonomiese ondersoek waardeur bronguskarsinoom identifiseer kan word. Nie-geoutomatiseerde sitologiese ondersoek is arbeidsintensief, tydrowend en vereis ‘n deurlopende hoë vlak van konsentrasie en fokus. Outomatisering is onlangs suksesvol geïmplementeer in ginekologiese sitologie-ondersoeke. Aangesien sputummonsters soortgelyk aan servikale monsters is, het die vraag ontstaan of sputummonsters ook geskik sou wees vir geoutomatiseerde sifting. Doelwit: Hierdie studie het verskeie doelwitte gehad: 1) Om geoutomatiseerde sifting van sputummonsters te toets deur gebruik te maak van BD Focal Point ™ Slide Profiler (FP), en te vergelyk met nie-geoutomatiseerde sputum sifting. 2) Om die sensitiwiteit en spesifisiteit van sputum in die identifikasie van bronguskarsinoom te bepaal. 3) Om vas te stel of enige kliniese, radiologiese of brongoskopiese bevindings bronguskarsinoom sou kon voorspel. 4) Om die belang van ‘n verteenwoordigende monster te bepaal. Metode: ‘n Prospektiewe studie van die pasiënte wat die Divisie van Pulmonologie by Tygerberg Hospitaal vir transbrongiale nodale aspirasie (TBNA) of ‘n transtorakale aspirasie (TTNA) vanaf Julie 2010 tot Mei 2012 bygewoon het, is gedoen. ‘n Prebrongoskopiese sputum is geneem en gestuur vir prosessering. Die gekleurde skuifies is deur die FP gestuur vir geoutomatiseerde ondersoek. Indien die sputumskuifies gekwalifiseer het vir geoutomatiseerde sifting, is hulle in die groep vir ondersoek ingesluit. ‘n Korrelasiestudie, om die sputumresultate te evalueer, is uitgevoer deur die TBNA/TTNA bevindings as standaard te gebruik. Resultate: Vir hierdie studie is 108 sputummonsters ingesluit. Vanuit die 84.3% maligne (n=91) en 15.7% benigne (n=17) gevalle, bevestig deur ‘n diagnostiese prosedure, het sputumsitologie ‘n sensitiwiteit van 38.5% (35/91 maligne gevalle) en ‘n spesifisiteit van 100.0% (17/17 benigne gevalle), getoon. Geoutomatiseerde sifting het ‘n beter sensitiwiteit met 94.3% (33/35 maligne gevalle), terwyl nie-geoutomatiseerde (ondersoek) ‘n sensitiwiteit van 74.3% (26/35 maligne gevalle) wanneer met die finale resultaat vergelyk, gevind. Individuele parameters met ‘n betekenisvolle assosiasie het die teenwoordigheid van ‘n endobrongiale tumor, gedeeltelike lugwegobstruksie / stenose, ronde massa, ‘n spekuleerde massa (negatiewe assosiasie), gewigsverlies (negatiewe assosiasie) en plaveiselkarsinoom as die histologiese subtipe, ingesluit. Geskiktheid van die monster was nie so betekenisvol as wat in die hipotese gestel is nie: aangesien 85.3% van ware positief gediagnoseerde sputummonsters, maar ook 65.5% van die vals negatiewe sputummonsters, groot hoeveelhede alveolêre makrofae ingesluit het. Gevolgtrekking: Sputumsitologie bly steeds ‘n belangrike deel van die siftingsprogram vir bronguskarsinoom in die openbare gesondheidssektor in Suid-Afrika. Resultate van hierdie studie bevestig dat sputumsitologie baie spesifiek is en dat geoutomatiseerde sifting die sensitiwiteit verbeter. Ge-outomatiseerde sifting het bewys dat dit meer tydsbesparend is, met ‘n 83.1% vermindering (p<0.0001) in die siftingstyd wat deur een sitotegnoloog per geval bestee word. Resultate het bevestig dat die hoeveelheid alveolêre makrofae nie direk proporsioneel verwant is tot die patologie nie. Hoe meer verteenwoordigend die sputummonster was, hoe groter was die kanse om ‘n akkurate positiewe diagnose te maak. Die assosiasie van die geskiktheid van die sputummonster en die positiewe resultate het egter nie ‘n statisties betekenisvolle resultaat getoon nie. Aanbevelings vir hierdie studie sluit in die aanwending van geoutomatiseerde sputumondersoeke.

The mitochondrion is a tiny organelle that is the power supplier of the cell and vital to the functioning of the body organs. Additionally it contains a small circular genome of about 16 kb, present in many copies which makes the mitochondrial DNA more viable than nuclear DNA. Mitochondrial DNA is also maternally inherited and thus provides a direct link to maternal relatives. These two properties are of particular use for forensic samples, which only contain limited or degraded amounts of DNA, and for historical samples (ancient DNA). This thesis presents work on the mitochondrial DNA in the hypervariable regions (HV) I and II, in both contemporary and historical samples. Forensic genetics makes use of mitochondrial DNA analysis in court as circumstantial evidence, and population databases are used for the calculation of evidence value. Population samples (299) across Sweden have been analysed in order to enrich the EDNAP mtDNA database (EMPOP) (paper I). The application of mitochondrial DNA analysis allowed for analysis of historical skeletal remains: Copernicus, 1473-1543 (paper II), Karin Göring, 1888-1931 (paper III) and Medieval bones, 880-1000 AD, from a mass grave found in Sigtuna, Sweden (paper IV). The thesis also includes analyses of bones and teeth from the shipwrecked crew of the Vasa warship, 1628, samples from the Vasa museum, Stockholm, Sweden (paper V). Overall, the varying age of the samples and the different conservation environments (soil and water) accounted for variations in quality, but still allowed for successful DNA analysis.

[pt] Métodos de análise para determinação de elementos-traço em amostras oleosas e pastosas são necessários, por exemplo, no caso do petróleo e em suas frações mais pesadas, para os quais os elementos-traço indicam características do tipo do petróleo, correlacionando-o com a localização geográfica da jazida. Além disso, a quantificação destes elementos tem utilidade nas interpretações geoquímicas relacionadas com maturidade térmica, correlação óleo-óleo e migração primária e secundária do óleo e na obtenção de informações que permitem a criação de estratégias para realizar o refino do óleo, prevenindo contaminação dos catalisadores, corrosão de equipamento e potencial dano ambiental. No caso de óleos e gorduras vegetais, a presença destes elementos, mesmo em baixas concentrações ((mi)g g-1), pode acelerar processos oxidativos que geram peróxidos, aldeídos, cetonas, ácidos e epóxidos, que além de causarem a rancidez prematura dos óleos produzem efeitos patológicos no sistema digestivo e potencializam a ação de alguns carcinogênicos. As técnicas espectrométricas baseadas no uso do plasma (ICP OES e ICP-MS) estão entre as mais utilizadas na determinação de elementos- traço nesses tipos de amostra. Porém, os procedimentos de preparação de amostra mais tradicionalmente usados são os que destroem a matriz das amostras, solubilizando-as em ácidos inorgânicos fortes. Neste trabalho, novas estratégias de preparação de amostras de petróleo, óleo diesel, asfaltenos, óleos comestíveis, manteiga e margarina foram desenvolvidas visando à utilização do tradicional procedimento de nebulização de solução para introdução de amostra no plasma. No caso do óleo diesel, a estratégia avaliada foi a de preparação de amostras na forma de emulsão com n-propanol. Nesse caso, um planejamento fatorial indicou que fatores importantes para a determinação de elementos refratários foram tanto a massa de amostra na composição da emulsão (até 25% em massa) quanto a acidificação da emulsão. As amostras de óleo cru, óleos vegetais e gorduras vegetais foram preparadas como microemulsão em n-propanol. Alternativamente, um procedimento de extração assistida por ultra-som foi desenvolvido. A emulsão de óleo cru com n-propanol mostrou ser um bom meio de estabilização dos analitos em solução. No caso da extração ácida assistida por ultra-som para óleo cru, um planejamento fatorial mostrou que os fatores importantes para a determinação de elementos-traço foram, o tempo de aquecimento da amostra, a concentração do ácido nítrico e a exposição ao ultra-som. Para as amostras de óleos e gorduras vegetais, a otimização das proporções de componentes na emulsão com n- propanol proporcionou boa estabilidade e homogeneidade das amostras de óleo de oliva e óleo de soja (6 % em massa de óleo) e para margarina e manteiga (5 % m/m de óleo). Para o método da extração ácida assistida por ultra-som, as condições otimizadas para a preparação da amostra permitiram também a análise por ICP-MS, cujos resultados são discutidos e comparados com os resultados obtidos por ICP OES. Para as amostras de asfaltenos, o procedimento proposto foi o de extração em HNO3 concentrado assistida por ultrasom. Nesse caso, os mesmos parâmetros otimizados para o óleo cru foram usados. Os parâmetros operacionais foram otimizados para obtenção dos parâmetros de mérito, sendo a aplicação dos métodos propostos confirmada por testes de recuperação além de comparação com resultados obtidos com procedimentos tradicionais para esse tipo de amostra. Assim, nessa tese, foram desenvolvidos métodos para a determinação de Cr, Mo, V e Ti em óleo diesel e em óleo combustível, Cd, Co, Cr, Fe, Mn, Mo, Ni, Ti, V e Zn em óleo cru, Fe, Ni e V em asfaltenos. Cd, Co, Cr, Cu, Mn e Ni foram determinados em óleos e gorduras comestíveis utilizando preparação de amostra como microemulsão em n-propanol. Já Al, Ca, Cd, Cu, Fe, Mg, Mn, Ni, V e Zn foram determinados após extração ácida assistida por / [en] Analytical methods for the determination of trace elements in oily and viscous samples are necessary, for example, in the case of crude oil and its heavy fractions, where trace elements indicate specific oil characteristics and the geographic location of the natural deposit. Moreover, the quantification of these elements is relevant for geochemical interpretations concerning thermal maturity, oil-oil correlations and primary and secondary oil migration. Knowledge on trace element characteristics of crude oil is also important in oil refining strategies to prevent contamination of catalysts as well as equipment corrosion, and to minimize environmental pollution. In the case of vegetal oils and fats, the presence of certain elements, even in low concentrations ((mi)g g-1); can speed up oxidative processes and premature rancidity of oils, which may cause pathological effects in the digestive system, including cancer. Spectrometric techniques based on inductively coupled plasma (ICP OES and ICPMS) are extensively used for determination of trace elements in such type of samples due to their sensitivities and multielemental features. In the case of oils and fats, the samples have to be generally decomposed by wet-ashing procedures with mineral acids, prior to the elemental determinations. In this work, new strategies for sample preparation of crude oil, diesel, asphaltenes, edible oils, butter and margarine have been developed, which allow the use of the traditional procedure for sample introduction into the ICP by solution nebulization. In the case of diesel oil, the proposed strategy was the preparation of samples as emulsions with n-propanol. Factorial design studies showed that the important factors for the quantitative determination of refractory metals were the sample amount in the emulsion (up to 25% in weight) and the acidification of the emulsion. Crude oil samples, vegetable oils and fats have been prepared as n-propanol microemulsions. Alternatively, an ultrasound-assisted trace metal extraction procedure was developed. The crude oil emulsions with n-propanol were found to be a good environment to keep the analytes in solution. In the case of the ultrasound-assisted acid extraction for crude oil, a factorial design indicated that the important factors for the determination of trace elements have been the sample heating time, the nitric acid concentration and the ultrasound exposure time. For vegetable oils and fats, the optimization of the npropanol emulsion components provided good stability and homogeneity olive and soy oils (6 % of oil in weight) and for margarine and butter (5 % of oil in weight). For the ultrasound-assisted acid extraction, the optimized conditions for sample preparation also allowed the analysis of the extracts by ICP-MS, which results were compared with the ones obtained by ICP OES. For asphaltene samples, the proposed procedure was the ultrasound- assisted acid extraction in concentrated HNO3 using similar conditions employed for crude oil. The figures of merit for the proposed methods were achieved under optimized experimental and instrumental conditions and validations were performed by analysis of certified reference materials (when available), recovery tests, and by comparison of results obtained using traditional sample preparation procedures for this type of sample. Methods for the determination of Cr, Mo, V and Ti in fuel and diesel oils, for Cd, Co, Cr, Fe, Mn, Mg, Ni, Ti, V and Zn in crude oil and for Fe, Ni and V in asphaltenes have been developed. Cd, Co, Cr, Cu, Mn and Ni in edible oils and fats have been determined using the microemulsification in n-propanol, while Al, Ca, Cd, Cu, Fe, Mg, Mn, Ni, V and Zn have been determined after acid extraction in HNO3. The analyses were carried out by using the external calibration method with internal standardization by Sc (in the case of ICP OES). The ultrasound-assisted acid extraction method for t

Submitted by Maria Aparecida (cidazen@gmail.com) on 2017-04-12T15:30:06Z No. of bitstreams: 1 Juliana Santos Farias.pdf: 1234303 bytes, checksum: 420e49b91acb05fb1346e95075411509 (MD5) / Made available in DSpace on 2017-04-12T15:30:06Z (GMT). No. of bitstreams: 1 Juliana Santos Farias.pdf: 1234303 bytes, checksum: 420e49b91acb05fb1346e95075411509 (MD5) Previous issue date: 2017-03-06 / CAPES, FAPEMA,CNPQ / This paper describes the development of a simple and new sensor electrochemical determination of flutamide in voltamétrica formulations pharmaceutical and artificial urine specimens using a carbon electrode vitreous (ECV) modified with oxidized carbon nanotubes and graphene oxide (NCO-OG), which was named ECV/NCO-OG. Electronic microscopy techniques scanning (SEM) and Raman Spectroscopy were used for the characterization of carbon based materials. The electrochemical response of the analyte front of the ECV/NCO-OG was investigated by cyclic voltammetry techniques (VC) and square wave voltammetry (VOQ). The sensor exhibited a high activity eletrocatalítica for the reduction of flutamide in 0.05 V vs. Ag/AgCl. The parameters experimental influence the response of the electrode was investigated and the optimum conditions were found for the electrode modified with NCO-OG, in Britton-Robinson buffer solution-BR on concentration of 0.1 mol L-1 (pH 5). The proposed sensor presented a wide range of linear response of concentration for the flutamide of 0.1 to 1000 µmol L-1 (or µ g L-1 27.6 to 0.27 g L-1) for n = 15 (R2 = 0.997), limit of detection (LOD), limit of quantification (LOQ), and sensitivity of 0.03 µmol L-1, 0.1 µmol L-1, and 0.30 µmol µA -1 L, respectively. The ECV/NCO-OG was successfully applied to the determination of flutamide in pharmaceutical formulations used in the treatment of prostate cancer and artificial urine samples. The results obtained with the proposed sensor were compared with the method described in the literature and showed a level of 95% confidence, demonstrating that there is no statistical difference between the method of reference and the method proposed. The addition and recovery studies show that the proposed method presents a satisfactory accuracy with average value of 101% recovery ( 1)%. for the fortified samples. / Este trabalho descreve o desenvolvimento de um simples e novo sensor eletroquímico para determinação voltamétrica de flutamida em formulações farmacêuticas e amostras de urina artificial empregando um eletrodo de carbono vítreo (ECV) modificado com nanotubos de carbono oxidado e óxido de grafeno (NCO-OG), o qual foi denominado ECV/NCO-OG. As técnicas microscopia eletrônica de varredura (MEV) e a Espectroscopia Raman foram utilizadas para a caracterização dos materiais à base de carbono. A resposta eletroquímica do analito frente ao ECV/NCO-OG foi investigada através das técnicas de voltametria cíclica (VC) e voltametria de onda quadrada (VOQ). O sensor exibiu uma alta atividade eletrocatalítica para a redução da flutamida em 0,05 V vs Ag/AgCl. Os parâmetros experimentais que influenciam a resposta do eletrodo foram investigados e as condições ótimas foram encontradas para o eletrodo modificado com NCO-OG, em solução tampão Britton-Robinson-BR na concentração de 0,1 mol L-1 (pH 5). O sensor proposto apresentou uma ampla faixa de resposta linear de concentração para a flutamida de 0,1 a 1000 µmol L-1 (ou 27,6 µg L-1 a 0,27 g L-1) para n=15 (R2=0,997), com limite de detecção (LOD), limite de quantificação (LOQ), e sensibilidade de 0,03 µmol L-1, 0,1 µmol L-1, e 0,30 µA µmol-1 L, respectivamente. O ECV/NCO-OG foi aplicado com sucesso para a determinação de flutamida em formulações farmacêuticas utilizadas no tratamento de câncer de próstata e amostras de urina artificial. Os resultados obtidos com o sensor proposto foram comparados com o método descrito na literatura e observou-se um nível de confiança de 95%, demonstrando que não há diferença estatística entre o método de referência e o método proposto. Os estudos de adição e recuperação mostram que o método proposto apresenta uma exatidão satisfatória com valor médio de recuperação de 101% ( 1) %. para as amostras fortificadas.

Ziel der vorliegenden Arbeit war es, Screening–Methoden zu prüfen, um auf einfache, kostensparende Weise und mit ausreichender statistischer Sicherheit festzustellen, ob der Erreger der Paratuberkulose (Mycobacterium avium ssp. paratuberculosis) in einer Herde vorhanden ist oder nicht. Dazu wurden zwei auf dem kulturellen Erregernachweis beruhende Verfahren, die Untersuchung individueller Kotproben auf der Basis von Stichproben und die Untersuchung von Umgebungskotproben, einem serologischen Untersuchungsansatz, dem Nachweis von MAP–Antikörpern nach Aufkonzentrierung in gepoolten Einzelmilchproben, gegenübergestellt. Als Referenzmethode diente die kulturelle Einzeltieruntersuchung aller über 24 Monate alten Tiere des jeweiligen Bestandes. In 20 Thüringer Milchviehherden mit bekannter Einzeltierprävalenz wurden 5063 Einzeltierkotproben, 200 Umgebungskotproben und 262 aufkonzentrierte Milchpools aus 4337 laktierenden Rindern untersucht. Zusätzlich wurde eine systematische retrospektive Stichprobenkalkulation (nStichprobe = 1458 Einzeltierkotproben) vorgenommen. Die Kultivierung der Einzeltierkotproben und der Umgebungskotproben erfolgte über 12 Wochen auf HEYM–Nährmedium mit anschließender Speziesidentifizierung durch PCR und Ziehl–Neelsen–Färbung. Die Umgebungskotproben wurden zu zwei Untersuchungszeitpunkten (Frühjahr und Sommer) an jeweils fünf Lokalisationen eines Betriebes entnommen: Abkalbebereich, Melkbereich einschließlich Vorwartehof, Laufbereich, Haupttriebweg, Übergang zum Kälberbereich. Die Untersuchung der Milchpools erfolgte nach vorheriger Aufkonzentrierung mittels zweier verschiedener ELISAs. Im Frühjahr entnommene Umgebungskotproben aus 16 MAP–positiven Betrieben detektierten das Vorhandensein des Erregers in neun Betrieben (56,3 %). Betriebe mit einer Einzeltierprävalenz von über 4,5 % wurden in neun von zehn Fällen (90 %) sicher erkannt. Im Sommer entnommene Umgebungskotproben fielen durch eine sehr starke Kontamination auf. Von den 16 MAP–positiven Beständen wurden 15 Herden (93,7 %) mittels Stichprobenuntersuchung als Bestand mit Paratuberkulosevorkommen identifiziert, wobei lediglich ein Bestand mit einer Einzeltierprävalenz von 0,49 % nicht detektiert wurde. Die serologische Untersuchung der Milchpools lieferte keine verwendbaren Ergebnisse. Mit Hilfe der Untersuchung von Umgebungskotproben lassen sich Herden mit einer durch kulturelle Untersuchung ermittelten Einzeltierprävalenz von 4,5 % und darüber mit hinreichender Sicherheit auffinden. Bei der Bewertung dieses Schwellenwertes ist zu beachten, dass bei Verwendung von nur einem Kulturröhrchen je Kotprobe von einer Sensitivität der Methode von 60 % im Vergleich zur Verwendung von drei Kulturröhrchen auszugehen ist. Für eine Überwachung unverdächtiger Herden ist die Sensitivität dieses Untersuchungsansatzes jedoch zu gering. Die individuelle kulturelle Untersuchung einer Stichprobe zeigte eine ausreichend hohe Sensitivität, um bei der Überwachung größerer unverdächtiger Herden eingesetzt werden zu können. Ein Einsatz serologischer Milchuntersuchung ist zur Bewertung von Beständen zur Ermittlung des MAP–Infektionsstatus gegenwärtig nicht zu empfehlen. Für die Überwachung größerer, als Paratuberkulose–unverdächtig anerkannter Bestände ist somit ein wechselnder Einsatz von Einzeltierkotproben aller Rinder über 24 Monaten und einer aussagekräftigen systematischer Stichprobenuntersuchung möglich und trägt damit zur Erleichterung der derzeit noch zeit– und kostenintensiven Paratuberkulose–Diagnostik bei.