Dissertations / Theses on the topic 'Interfering methods'

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis complex species, which infect the exposed individuals, initiate an immune response with production of several cytokines, among them interferon-gamma (IFN-gamma), which is crucial for activation of cellular immune response against TB bacillus. This cytokine has been extensively studied as a possible biomarker for diagnosis of TB. In this work, we analyzed the levels of IFN-gamma in response to in vitro stimulation with ESAT-6, CFP10 and TB7.7, by QuantiFERON ®-TB Gold (In-tube method) (QTF-IT) in 155 participants: 47 patients confirmed pulmonary TB (patient group), 49 direct contacts (group contact), and 59 individuals without history or contact with TB (control group). The levels of IFN-gamma were different among the groups (p=0.0001); the highest level was observed in the patients group (median=1.43 IU/mL). In contacts group, 20/49 (40.8%) subjects were positive for the QTF-IT (median = 0.26 IU / mL), and among these positive contacts 14/20 (70.0%) exhibited high levels of IFN-gamma, (≥ 1.05 IU/mL). The performance testing of TB patients versus controls, resulted in 80.9% sensitivity (95% CI = 69.6% to 92.1%) and 93.2% specificity (95% CI = 86.8 % to 99.6%), but when the results of contacts and control group were combined, the specificity decreased to 77.8% (95% CI = 69.9% to 85.6%). Thus, according to our data, there is a distinctive profile of IFN-gamma, with higher levels in the patients group. The QTF-IT positive in contacts, especially those with high levels of IFN-gamma, should be monitored, especially those with high levels of IFN-gamma. This result may be a risk factor for developing TB disease. In addition, the QTF-IT, properly executed, can be useful in endemic areas for the diagnosis of latent and active TB (infection) when evaluated with other routine tests.
A Tuberculose (TB) é uma doença infecciosa causada por espécies do complexo Mycobacterium tuberculosis, que ao infectar os indivíduos expostos, iniciam uma resposta imunológica com produção de várias citocinas, dentre elas o Interferon-gama (IFN-gama), que é crucial para ativação da resposta imune celular contra o bacilo da TB. Esta citocina tem sido amplamente estudada como um possível biomarcador para auxiliar no diagnóstico da TB. Neste trabalho foram analisados as concentrações de IFN-gama em resposta a estímulos in vitro com ESAT-6, CFP10 e TB7.7, utilizando o QuantiFERON®-TB Gold (In-tube method) (QTF-IT), em 155 participantes, sendo: 47 pacientes com TB pulmonar confirmada (grupo paciente), 49 contatos de pacientes com TB pulmonar (grupo contato), e 59 indivíduos sem história e/ou contato com TB (grupo controle). As concentrações de IFN-gama foram diferentes entre os grupos avaliados (p=0,0001), sendo mais elevado nos pacientes com TB (Mediana = 1.43 UI/mL). Dos contatos diretos, 20/49 (40,8%) foram positivos para o QTF-IT (Mediana=0,26 UI/mL), e destes 14/20 (70,0%) apresentaram níveis elevados de IFN-gama, superior à 1,05 UI/mL. O desempenho do teste relacionando os pacientes de TB como os doentes e os controles como sadios, resultou em 80,9% de sensibilidade (IC 95% = 69,6% a 92,1%) e 93,2% de especificidade (IC 95% = 86,8% a 99,6%), mas quando os resultados dos contatos diretos foram adicionados aos controles, a especificidade reduziu para 77,8% (IC 95% = 69,9% a 85,6%). Concluiu-se que há um perfil diferenciado na produção de IFN-gama, com níveis mais elevados nos pacientes de TB ativa, e que os contatos positivos para o QTF-IT, principalmente, aqueles com altas concentrações de IFN-gama, devem ser monitorados, considerando que o perfil ou concentração elevado desta citocina, pode ser um fator de risco para o desenvolvimento da TB doença. Além disso, o QTF-IT, executado corretamente, pode ser útil em áreas endêmicas para auxiliar no diagnóstico da TB latente (infecção) e ativa, quando avaliados com outros exames de rotina.

Manipulation of a virus's capacity to circumvent the interferon (IFN) response aids both fundamental studies as well as many practical applications including the design of live-attenuated vaccines. However, these IFN-sensitive viruses are often difficult to grow to high titer in cells that produce and respond to IFN. In the first part of this study we further characterised the use of the IFN inhibitor, Ruxolitinib (Rux) for its ability to block the IFN response and subsequently enhance replication of IFN-sensitive viruses. This study has shown that i) Rux could provide a more rapid and therefore more efficient alternative for the growth of IFN-sensitive viruses than the current default option, growth in Vero cells and ii) addition of Rux can increase growth of multiple viruses in numerous cell-lines. These results indicate that as well as aiding fundamental studies the addition of Rux could become a valuable technique in a number of virological applications including live attenuated vaccine production and techniques to isolate newly emerging viruses. In the second part of this study we developed a novel method to isolate IFN-sensitive viruses from Paramyxoviruses, using PIV5 (Parainfluenza virus 5) as an experimental model system to obtain selection parameters. We successfully isolated three mutant viruses (rPIV5mCh-α, rPIV5mCh-β and PIV5 W3-γ) that each contain mutations within the IFN antagonist V protein and the P protein which is essential for RNA replication. Subsequently, both rPIV5mCh-α and PIV5 W3-γ were shown to contain non- functional V proteins and exhibit IFN-sensitivity. Ultimately, this study is the first step towards creating a general method to isolate various types of IFN-sensitive viruses that as well as aiding fundamental studies, may be further developed as attenuated vaccines for clinically important viruses lacking vaccines.

Background Multiple sclerosis (MS) is a chronic inflammatory demyelinating disorder of the central nervous system (CNS). In Hong Kong, the prevalence rates of MS is 4.8/100,000. First line disease modifying agent (DMA) type 1 interferon β (IFN-β) is the most commonly use therapy for relapsing and remitting MS(RRMS). Depending on the administration type and route of IFN-β, up to 80% of patients develop harmless binding antibody (BAb),which binds to IFN molecules but not necessary interfere its bioactivity. When IFN-β therapy continues, maturation of BAb response can lead to the formation of high affinity neutralizing antibody (NAb). About 45% of MS patients develop NAb against IFN-β in one year of IFN-β treatment. NAb shows a loss of IFN-β clinical effect by increasing MRI activity and disease progression. As the clinical effect of NAb is lagging behind the initial appearance of NAb in the body, it is suggested to develop a NAb assay to predict treatment failure and advice switching therapy for patients when NAb is present. Aim The aim of this study was: I. To develop and evaluate a qPCR-based method for the measurement of NAb to IFN-β in MS patient. II. To establish the normal reference range of NAb in Chinese population. III. To seek the possibility of using anti-IFN-β BAb assay and in vivo MxA gene expression assay as a screening test for NAb IV. To compare the performance between MxA induction qPCR, ELISA, WB assay and luciferase IFN-β reporter gene assay Materials and methods23Chinese RRMS patients who treated with IFN-β-1a therapy for a minimum of12 months were recruited in this study. Serum and PBMC were collected12 hours after the IFN-β-1a injection. MxA, IFNAR1 and IFNAR2 mRNA from PBMC were tested byin vivo MxA gene expression assay. NAb containing serum was tested by anti-IFN-β BAb assay, IFN-β reporter gene assay, in vitro MxA induction WB, ELISA and qPCR assay. In addition, blood samples from 3 Chinese volunteers without any known autoimmune disease history were collected to evaluate the baseline of NAb titer and MxA expression. Result The experimental condition of MxA induction qPCR assay was optimized by using 2.5×105A549 cells plating density, 10% FCS concentration,5 hours IFN-β stimulation time and GAPDH normalization. Assay accuracy was validated by reference anti-IFN-β antibody. Starting from 2.5 TRU, linear relationship could be observed (r2= 0.9873). The lower limit of quantification (LLOQ) was 0.02 LU/mL, the upper limit of quantification (ULOQ) was 16LU/mL and the limit of detection (LOD) was 0.002 LU/mL. The reproducibility of the assay was measured, the intra-and inter-assay imprecision(%CV)for high value were 5.95% and 7.17% respectively, while the intra-and inter-assay impression were8.31% and 15.95%respectively.Results of the qPCR-based method were concurring with that of luciferase IFN-β reporter gene assay. The upper limit of the NAb reference range in Chinese population was 40.3 TRU (n=3, 95% CI = 31.7-48.8). The performance observed in MxA induction ELISA assay swas unsatisfactory. The correlation of anti-IFN-β BAb assay and in vivoMxA gene expression assay results with NAb status indicated both tests were sensitive enough for NAb screening. Conclusion A normal range of NAb titer in Chinese population was established in this study. Anti-IFN-β BAb assay and in vivo MxA gene expression assay were proved suitable for NAb screening. The performance of the developed MxA induction qPCR assay was superior to MxA induction ELISA, WB assay and comparable to luciferase IFN-β reporter gene assay. By using MxA induction qPCR assay, actual efficacy of IFN-β therapy could be measured and monitored. Any treatment failure could be predicted earlier.
published_or_final_version
Pathology
Master
Master of Medical Sciences

85 f.
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CAPES
O objetivo deste trabalho foi o desenvolvimento de um procedimento analítico empregado Cromatografia Líquida Ultra Rápida acoplada a Detector de Fluorescência (UFLC-FLU) para determinação de interferentes endócrinos (IEs; bisfenol A (BPA), 4n-nonilfenol (4NP), 4-octifenol (4OP), 4-t-octifenol (4TOP), estriol (E3), estrona (E1), 17β- estradiol (E2) e 17α-etinilestradiol (EE2)) em água do mar. Foi utilizado um sistema de pré-concentração em fase solida (cartucho SPE com fase estácionária C18) para extração e pré-concentração dos IE em as amostras de água do mar. A separação foi otimizada e realizada em um tempo total de corrida de 10 min, em uma coluna cromatográfica Shim-pack XR-ODS C-18 (2,0 mm ID x 50 mm), com a fase móvel de acetonitrila e água ultra pura com gradiente de eluição. A vazão foi de 0,12 mL min-1, a temperatura da coluna foi mantida em 60°C e os comprimentos de onda de emissão e excitação foram de 306 nm e 280 nm, respectivamente. O método validado foi aplicado em amostras de água coletadas na Baia de Todos os Santos, Bahia, Brasil. As amostras foram coletadas na Baía da Ribeira, Feira do São Joaquim, Santo Amaro da Purificação, São Francisco do Conde, Cachoeira e Acupe. As recuperações para o IE variaram entre 84,9% (para o composto 4nOP) e 104% (para o composto 4nNP), e a repetibilidade foi adequada (RSD < 4,5%). Os limites de detecção e quantificação encontrados para os compostos estudados variaram de 4 a 27 µg L-1 e de 19 a 185 µg L-1, respectivamente para o método cromatográfico. Considerando o fator de concentração de 2000 vezes, o LD e LQ variaram de 2 a 23 ng L-1 e de 9 a 96 ng L-1, respectivamente quando calculados para amostra real. Foi observada a ocorrência dos seguintes IE nas amostras reais: bisfenol A (BPA), 17β-estradiol (E2), estriol (E3) e 4n-octifenol (4NOP) em concentrações que variaram de 5 ng L-1 em Santo Amaro a 18,3 ng L-1 em Cachoeira para o E2, 20 ng L-1 do 4NOP na Ribeira a 135 ng L-1 no estuario do Rio Subaé (Santo Amaro), sendo o 4NOP o contaminante onipresente nas amostras analisadas. A concentração de 38 ng L-1 de E3 foi encontrada apenas nas amostras da Ribeira. A presença de BPA foi detectada em quase todas as amostras (o BPA não foi detectado na Ribeira), em níveis entre 13 ng L-1 no estuário do rio Paraguaçu em Cachoeira e aproximadamente 77 ng L-1 no estuário do rio Subaé. Os resultados sugerem que as regiões estudadas encontram-se possivelmente impactadas em relação os IE estudados e que as concentrações encontradas poderiam indicar possíveis danos ao ecossistema marinho local. O método analítico empregando SPE e UFLC-FLU se mostrou eficiente na determinação dos oito compostos de interesse usando um volume de amostra de 4 litros.
Salvador

Biotin är ett vitamin som finns naturligt i livsmedel och det dagliga intaget nås via födan. Höga doser biotintillskott samt höga doser biotin i läkemedel, kan leda till biotininterferens i kliniska immunokemiska analyser. Roche Diagnostics® vill införa nya reagens med högre tröskel för biotininterferens för att minska risken för biotininterferens vid analys av patientplasma. Därför var syftet med studien att metodverifiera fyra nya reagens från Roche Diagnostics® som används vid diagnostisering och behandling av hjärtsvikt, sepsis, och prostatacancer. De fyra reagensen, Elecsys® proBNP II, Elecsys® BRAHMS PCT, Elecsys® total PSA samt Elecsys® free PSA metodverifierades för att användas på Cobas® e801. Studiematerialet bestod av 20 patientprover av litiumheparinplasma per reagens (totalt 80 patientprover). Resultatet av verifieringen av Elecsys® proBNP II visade en korrelation till det befintliga reagenset på r = 0,9998 och Bland-Altman analys visade en spridning av resultaten på < 10 %; inomserieprecisionsstudien gav CV 1,56 %. Elecsys® BRAHMS PCT hade en korrelation på r = 0,9997 och Bland-Altman analysen visade en spridning på > 10 %; inomserieprecisionsstudien gav CV 1,70 %. För Elecsys® total PSA och free PSA fanns korrelationen till det befintliga reagenset på r = 1 respektive 0,9997 och Bland- Altman analysen visade en spridning på < 10 % hos båda reagensen. Inomserieprecisionsstudien gav CV 0,44 % respektive CV 2,67 %. Resultaten för samtliga reagens uppvisar god korrelation till det befintliga reagenset och en hög mätnoggrannhet vilket talar för att de fyra nya reagensen kan tas i bruk.
Biotin is naturally found in foods, and we obtain this vitamin through our daily diet. Biotin supplements as well as high doses of biotin in drugs can lead to biotin interference in clinical immunochemical analyzes. Therefore, the purpose of this study was to methodically verify four new reagents from Roche Diagnostics® with a higher threshold for biotin interference, used in the diagnosis and treatment of heart faliure, sepsis and prostate cancer. The four reagents, Elecsys® proBNP II, Elecsys® BRAHMS PCT, Elecsys® total PSA and Elecsys® free PSA were method-verified for use on Cobas® e801. The study material consisted of 20 patient samples of lithium heparin plasma per reagent. In total 80 samples were analyzed.The result of the verification of Elecsys® proBNP II showed a correlation to the existing reagent of r = 0.9998 and Bland-Altman analysis showed a distribution of the results of <10 %. The withinseries precision study yielded CV 1.56 %. Elecsys® BRAHMS PCT had a correlation of r = 0.9997 and the Bland-Altman analysis showed a distribution of > 10 %. The withinseries precision study gave CV 1.70 %. For Elecsys® total PSA and free PSA, the correlation to the existing reagent was r = 1 and 0.9997, respectively, and the Bland-Altman analysis showed a distribution of <10 % in both reagents. The withinseries precision study yielded CV 0.44 % and CV 2.67 % respectively.The results for all reagents show a good correlation to the existing reagent and a high accuracy of measurement, which indicates that the four new reagents can be used.

Des stratégies de thérapie génique sont en cours d'étude pour le traitement de maladies humaines. Ce type de thérapie s'adresse en premier lieu aux anomalies héréditaires récessives causées par la déficience en un gène unique dont le traitement nécessite l'addition du gène normal, mais pourrait être utilise également pour le traitement de cancers, de leucémies ou de maladies infectieuses. Le recours aux modèles animaux est indispensable pour : 1) juger directement de l'efficacité du transfert de gènes in vivo; 2) explorer les effets biologiques et potentiellement thérapeutiques de molécules exprimées à des niveaux physiologiques ou non physiologiques; 3) améliorer la méthodologie qui sera finalement appliquée à l'homme. Nos travaux s'inscrivent précisément dans le cadre du développement et de l'amélioration de modèles animaux murins. Étant donné les propriétés antivirales de l'interféron (IFN), nous développons une approche de thérapie génique basée sur l'expression constitutive de l'IFN dans les cellules hématopoïétiques et dirigée contre les maladies virales fatales à évolution lente et chronique comme le sida. D'autre part, nous nous intéressons à une maladie génétique héréditaire causée par la déficience en un gène unique codant pour l'adénosine déaminase (ADA); le dysfonctionnement de ce gène entraine une immunodéficience combinée sévère (SCID) chez l'homme. Aussi, la seconde partie de nos travaux concerne l'amélioration de la technologie relative à l'expression du gène codant pour l'ADA humaine dans les cellules hématopoïétiques in vivo, dans un modèle murin. L’approche commune à ces deux projets repose sur l'expression des ADNc (ADN complémentaires) des gènes codant pour l'IFN-beta murin et l'ADA humaine dans les cellules hématopoïétiques par l'intermédiaire de vecteurs rétroviraux.

碩士
國立中央大學
環境工程研究所
96
The thermal-optical method is widely used for measuring atmospheric aerosol carbons. In this method, pyrolyzed carbon correction can be split into thermal optical reflectance (TOR) and thermal optical transmittance (TOT). This study compares the differences due to different temperature protocols and pyrolyzed carbon corrections. Meanwhile, the interferences of volatile organic carbons (VOCs) on aerosol carbon analysis for filter samples are also studied. Moreover, the effects of NaCl on EC fractionization in the coastal area are evaluated. Finally, the monitoring data of aerosol black carbon (BC) and elemental carbon (EC) from North aerosol supersite are adopted to conduct a linear regression analysis. The slope resulted from this analysis is considered to be the best BC absorption cross section (σ) and the influencing factors of σ values are also disussed. The results show that TOR tends to underestimate pyrolyzed carbon correction when atmospheric carbon concentration is increased and thus the depth of particle deposition on a filter. Under the influence of vehicle exhausts and ambient temperature, VOCs adsorption by quartz fiber filter will be increased when using the filter in aerosol collection. Meanwhile, organic carbon evaporation from deposited particles is severer. The interference of Na+ on EC fractionization is significant when Na+ concentration reaches 3 - 6 μg m-3 based on the observation at the Shimen site on the coastline of Taipei County. In this study, 17,000 hourly data from North aerosol supersite were used to obtain the correction value of σ. The variation of σ is within the range of 20 - 40 m2 g-1 during this time period. Meanwhile, the value of σ is found to increase with higher aerosol nitrate, higher ozone concentration, higher atmospheric relative humidity above 90%, and the duration of raining event. In the diurnal variation of σ, σ value is found to enhance in the traffic peak hours. The diurnal variations of σ value are also agreed well with that of PM2.5, PAH, and OC. It indicates that the increase of atmospheric aerosols and aerosol organic fractions will affect σ values. The best σ value in the greater Taipei metropolis is recommended to change from the manufacturer’s 16.6 to 24.5 m2 g-1 for AE31 Aethalometer in measuring BC at 880 nm wavelength.

Indiana University-Purdue University Indianapolis (IUPUI)
MicroRNAs (miRNA) are short, 18-24 nucleotide long noncoding RNAs. These small RNAs, which are initially transcribed in the nucleus, are transported into the cell cytoplasm where they regulate protein translation either through direct cleavage of mRNA, or indirect inhibition through binding to mRNA and disrupting the protein translation machinery. Recently, miRNAs have gained much attention due to their implication in numerous diseases and cancers. It has been found that heightened or lowered levels of miRNA in diseased cells vs. healthy cells are linked to disease progression. It is therefore immensely important to be able to detect these small molecules. Current detection methods of Northern blotting, microarrays, and qRT-PCR suffer from drawbacks including low sensitivity, a lack of simplicity, being semi-quantitative in nature, time-consuming, and requiring expensive instruments. This work aims to develop novel miRNA technologies which will address these above problems. Bioluminescent labels are promising alternatives to current methods of miRNA detection. Bioluminescent labels are relatively small, similar in size to fluorescent proteins, and they emit very intense signals upon binding to their substrate. Bioluminescent labels are advantageous to fluorescent labels in that they do not require an external excitation source, rather, the excitation energy is supplied through a biochemical reaction. Therefore, background signal due to excitation is eliminated. They also have the advantage of being produced in large amounts through bacterial expression. Four miRNA detection methods are presented which utilize luminescence-based methods. Three employ Renilla luciferase, a bioluminescent protein, and one is based on fluorescence. The presented methods are capable of detecting miRNA from the picomole (nanomolar) level down to the femtomole (picomolar) level. These methods are rapid, sensitive, simple, and quantitative, can be employed in complex matrices, and do not require expensive instruments. All methods are hybridization-based and do not require amplification steps.

Selective gas-phase dissociation strategies have been developed for the characterization of cross-linked peptides and proteins in quadrupole ion trap mass spectrometers. An infrared chromogenic cross-linker (IRCX) containing a phosphotriester afforded rapid differentiation of cross-linked peptides from unmodified ones in proteolytic digests of cross-linked proteins by selective infrared multiphoton dissociation (IRMPD). Only the cross-linked peptides containing the chromogenic phosphate underwent IRMPD and unmodified peptides were not affected by IR irradiation. IRMPD of IRCX-cross-linked peptides yielded uncross-linked y-ion sequence tags of the constituent peptides due to secondary dissociation of all primary product ions which contained the chromophore, thus allowing successful de novo sequencing of the cross-linked peptides. Peptides cross-linked via a two-step conjugation strategy through the formation of a bis-arylhydrazone (BAH) bond were selectively dissociated by ultraviolet radiation at 355 nm. The BAH-cross-linked peptides could be distinguished from not only unmodified peptides but also dead-end modified peptides based on the selectivity of ultraviolet photodissociation. In a complementary approach, electron transfer dissociation of BAH-cross-linked peptides resulted in preferential cleavage of the hydrazone bond which produced two modified peptides. These modified peptides were subsequently interrogated by CID which allowed for the original site of cross-linking to be pinpointed. IRMPD was implemented in a dual pressure linear ion trap to demonstrate successful photodissociation of peptides having modest absorptivities. Peptides were observed to efficiently dissociation by IR irradiation exclusively in the low pressure cell whereas no dissociation was observed in the high pressure cell due to extensive collisional cooling. IRMPD provided greater sequence coverage of the peptides than CID and yielded product ion mass spectra which were predominantly composed of singly charged product ions which simplified spectral interpretation. IRMPD was further applied for the sequencing of small-interfering RNA. Complete sequence coverage was obtained and the results were compared to CID.
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Rozprawa odnosi się przede wszystkim do zaburzeń artykulacji, które, podobnie jak wszystkie inne nieprawidłowości, ograniczają człowieka. Wady wymowy stanowią bowiem poważny problem już we wczesnych latach życia, gdy dziecko bywa wyśmiewane ze względu na swoje problemy, a także w okresach późniejszych, stanowiąc przeszkodę w pełnieniu obowiązków lektora, aktora, prawnika czy nauczyciela. Wadliwa artykulacja, wbrew powszechnej opinii, nie jest najprostszym zaburzeniem mowy, z jakim mierzy się logopeda. Doświadczenia praktyków pokazują jednoznacznie, że terapia wskazanych tu nieprawidłowości w niektórych przypadkach jest bardzo trudna i rozciągnięta w czasie, dlatego istnieje tak wielka potrzeba stworzenia nowego podręcznika na temat kształtowania i automatyzacji głosek języka polskiego, którego wyraźnie brakuje po opublikowanym w 1967 roku „Jak usuwać seplenienie i inne wady wymowy” D. Antos, G. Demel i I. Styczek. Pierwsze rozdziały rozprawy doktorskiej, stanowiące część teoretyczną, zawierają rozważania na temat systemu fonologicznego języka polskiego, normy wymawianiowej, zaburzeń artykulacji, właściwości mowy dziecięcej oraz metody i techniki pracy z dziećmi z dyslalią.W drugiej części pracy, w której zamieszczono opis badań własnych, omówione zostały zagadnienia metodologiczne przeprowadzonego eksperymentu, tj. cele i przedmiot badań, założenia terminologiczne oraz podstawy ujęcia statystycznego wyników. W kolejnych rozdziałach przedstawiono analizę wyników badań na temat dziecięcego poczucia poprawności językowej, wyniki badań eksperymentalnych, metodykę postępowania logopedycznego w poszczególnych grupach badawczych oraz propozycję artykulacyjnego minimum sprawnościowego. Głównym zagadnieniem poruszonym w pracy jest metodyka wywoływania i utrwalania głosek w terapii dzieci z dyslalią obwodową. Poddane eksperymentowi techniki terapeutyczne zostały podzielone na potrzeby przeprowadzonego badania na dwie grupy metod, nazywanych tu ingerującymi i nieingerującymi. Głównym kryterium takiego podziału była stymulacja dotykowa pacjenta lub jej brak. Celem teoretycznym przeprowadzonego eksperymentu było poznanie wpływu metod ingerujących (zakładających kontakt dotykowy logopedy z pacjentem) i nieingerujących (bez kontaktu dotykowego) na poprawę umiejętności artykulacyjnych dzieci w wieku przedszkolnym. Badania miały również odpowiedzieć na pytanie, czy dostarczenie większej ilości bodźców zmysłowych (dotykowych, czuciowych) wpływa na skuteczność prowadzonej terapii wad wymowy. Praktyczne wnioski płynące z badań wiążą się z dostarczeniem niezbędnej wiedzy na temat właściwego wykorzystywania określonych metod i technik terapeutycznych w procesie wywołania normatywnych głosek języka polskiego. Eksperyment miał zatem odpowiedzieć na pytanie, jakie działania powinien podjąć terapeuta, by proces poprawy artykulacji był szybszy i efektywniejszy. Przedstawione wyniki ogólne potwierdzają zasadność postawionej na początku pracy tezy o wyższej skuteczności terapii wad wymowy dzięki wykorzystaniu metod i technik ingerujących. Zaprezentowane w pracy próby warg i języka stanowią podstawę artykulacyjnego minimum sprawnościowego, w obrębie którego znajdują się także ćwiczenia ukierunkowane tylko i wyłącznie na terapię określonej głoski lub grupy głosek. Za podstawę uzyskania dźwięku uznawano zatem pełen zasób prób warg i języka oraz określone ruchy dedykowane prowadzonej korekcji mowy.Badania miały za zadanie wykazać, że umiejętnie stosowane metody ingerujące są skutecznym sposobem na przyspieszenie i uatrakcyjnienie terapii. Dzięki powstającym ciągle pomocom logopedycznym, opartym na wnikliwej analizie ich skuteczności, możliwe jest stosowanie metod zgodnych z zasadami sensomotoryki człowieka, które dążą do celu ostatecznego – poprawy funkcjonowania pacjenta w społeczeństwie.
The work primarily refers to disorders of articulation, which, like all other abnormalities, restrain a human being. Speech defects constitute a serious problem already in the early years of life, when a child is sometimes ridiculed because of its problems, and also in later periods, obstructing the performance of duties of a reader, an actor, a lawyer or a teacher.Defective articulation, contrary to popular belief, is not the easiest speech disorder, faced by a speech therapist. Practitioners' experience clearly demonstrate that therapy of the abnormalities indicated here, in some cases, turns out to be very difficult and stretched in time, which is why there is so great a need for a new manual on shaping and automation of the sounds in the Polish language, which is clearly missing after "How to remove lisp and other speech defects" by D. Antos, G. Demel and I. Styczek, the published in 1967.The first chapters, constituting the theoretical part, contain considerations on the Polish phonological system, standards of pronunciation, articulation disorders, properties of children's speech, and methods of dealing with peripheral dyslalia. The next section discusses the methodological issues of the experiment conducted, i.e. the objectives and the target of the research, the terminology assumptions and the basics of statistical presentation of results. The third and fourth chapters present detailed findings of the research and the minimum articulation skill.A description of the results of the experiment was divided into three main parts: the detailed results, the aggregate results of all the research groups and the methodology of logopedic conduct during the therapy of individual speech defects. The above sequence of discussion is justified primarily by the fact that differences in the higher efficacy of interfering methods over the non-interfering ones grow in direct proportion to the increase in the level of generalization of results, i.e. the greater the data range, the higher the statistical significance confirming the thesis formulated in this dissertation.The main focus of the work is the methodology of developing and saving sounds in the treatment of children with peripheral dyslalia. The therapeutic techniques subjected to the experiment were divided for the purpose of the research conducted into two groups of methods, referred to as interfering and non-interfering here. The main criterion for such a division was the patient's tactile stimulation or its lack.The theoretical aim of the conducted experiment was the exploration of the impact of the interfering and non-interfering methods on the correction of articulation skills in preschool children. The research was also supposed to answer the question of whether providing more sensory stimuli (tactile, sensory) affects the effectiveness of the therapy of speech defects.The practical conclusions from the research are associated with delivering the necessary knowledge about the proper use of specific methods and therapeutic techniques in the process of calling normative Polish language sounds. The experiment was therefore to answer the question of what action should be taken by a therapist to make the process of articulation improvement more efficient and more effective.Exercises to lips and tongue presented in this work, form the basis of minimum articulation skills, within which there are also exercises targeted exclusively to the treatment of specific sound or group of sounds. A full resource of lips and tongue trials and specific movements dedicated to the conducted speech correction is therefore considered as the basis for obtaining a sound.

碩士
國立臺灣大學
海洋研究所
101
This study investigates effects of temperature, iodide concentration, oxidants and suspended solid on the colorimetric determination of oxygen in natural waters. Since the measurement involves the mixture of two iodine species (i.e. I2 and I3-), the equilibrium between them becomes a crucial factor to the precision of the method. A high concentration of iodide in the pickling reagent was found essential to ensure a stable [I_3^- ]/[I_2 ] ratio. Besides, it was also found that elevated temperature can cause a lower signal of molecular iodine, but increase the absorbance of triiodide ion. An over-all increment of 0.4% per 1 ℃ has been observed for a freshwater sample having oxygen concentration of ca. 250 μM. Therefore, temperature deviation can not only change the equilibrium constant but also alter the spectra of both iodine species. To solve this problem, the spectrophotometer has been modified using a thermostat unit which ensures the temperature difference between samples be kept within 0.5 ℃. Iodate, hydrogen peroxide, nitrite which are known to exist in natural environment all interfere the oxygen measurement. A quick reverse pickling procedure can be used to identify the iodate interference. Addition of sodium azide is always suggested to remove overestimate from nitrite. Since the sampling for oxygen usually excludes filtration therefore suspended solid in a sample can be another source of error. Re-measurement of the absorbance of the same sample after adding sodium thiosulfate reagent to remove the iodine color is considered the most effective way to correct for the interference from turbid water.