My dissertation is primarily divided into two parts. The first deals with the preparation, functionalization, and characterization of carbon surfaces prepared by direct current magnetron sputtering (DCMS) and high power impulse magnetron sputtering (HiPIMS) as substrates for bioarrays. Part two discusses applications of XPS peak fitting in surface chemical analysis. Chapter 1, the introduction, includes (i) a discussion of the construction of bioarrays and the preparation of sputtered surfaces, e.g., by DCMS and HiPIMS, and also functionalization (bioconjugate) chemistry with special emphasis on the importance of covalent functionalization of surfaces, and (ii) a discussion of the surface characterization techniques and accompanying analysis methods I have primarily used, which include X-ray photoelectron spectroscopy (XPS), near-ambient pressure XPS (NAP-XPS), XPS peak fitting, and contact angle goniometry (wetting). Chapter 2 discusses the preparation, characterization, and functionalization of DCMS and HiPIMS carbon surfaces for bioarrays. Here, two functionalization chemistries are explored, where the activity of DCMS and HiPIMS carbon towards amidation and amination is compared. Chapter 3 focuses on the use of Gaussian-Lorentzian sum (GLS) and Gaussian-Lorentzian product (GLP) line shapes in the context of peak fitting XPS narrow scans. This discussion includes a comparison of the GLS and GLP line shapes with the Voigt function. Chapters 4 and 5 discuss the applications of XPS peak fitting in materials characterization. Chapter 4 talks about XPS data analysis in the context of the chemical vapor deposition of various aminosilanes and their effect on peptide stability and purity. Chapters 5 describes the surface chemical analysis of various materials by NAP-XPS, including accompanying data analysis and/or peak fitting. The materials probed here cannot be analyzed at ultra-high vacuum by conventional XPS, hence, they are analyzed by NAP-XPS. Chapter 5 is divided into 5 sections. Section 5.1.1 discusses the characterization and analysis of a solution of bovine serum albumin (BSA) by peak fitting the C 1s and O 1s peak envelopes. Section 5.1.2 discusses the analysis of polytetrafluoroethylene (PTFE) at different pressures. Here, the effect of increasing background pressure and X-ray illumination time on the equivalent widths of the F 1s narrows scans is shown. Environmental charge compensation is also discussed here. Section 5.1.3 includes the analysis of poly(γ-benzyl L-glutamate) (PBLG), where the C 1s and O 1s peak envelopes were peak fitted to determine/confirm the structure and composition of this polymer. Section 5.1.4 contains an analysis and comparison of three different human hair samples: (i) untreated, (ii) colored, and (iii) bleached. Here, a comparison of the Si 2p, S 2p, and C 1s peaks illustrates the effects of the different treatments. Section 5.1.5 shows the characterization and analysis of liquid and solid phosphate buffered saline (PBS). Chapter 6 presents conclusion of my work and discusses future work.

Es ist bekannt, dass eine diabetische Stoffwechsellage über einen gesteigerten Proteinabbau zu einer Muskelatrophie führen kann. Ein zentrales System beim Abbau von Muskelproteinen ist hierbei das Ubiquitin-Proteasom-System mit seinen zwei spezifischen E3-Ligasen MuRF-1 und MAFbx. Ziel dieser Arbeit war es, den Einfluss einer Hyperglykämie auf die Expression von Proteinen des Ubiquitin-Proteasom-Systems im Skelett- und Herzmuskel zeit- und konzentrationsabhängig zu untersuchen. Insbesondere stand die Expression der E3-Ligasen MuRF-1 und MAFbx sowie die daraus folgende Auswirkung auf die Protein-Ubiquitinylierung im Fokus der Untersuchungen. Weiterhin sollte der Einfluss der Hyperglykämie auf die Apoptoserate von Skelett- und Herzmuskelzellen analysiert werden. Seit Kurzem stehen die GLP-1-Analoga als neue Antidiabetika für die Therapie des Diabetes mellitus Type II zur Verfügung. Da in Studien bereits positive Effekte der GLP-1-Analoga am Herzmuskel festgestellt wurden, sollte in der vorliegenden Studie geprüft werden, ob das GLP-1-Analogon Liraglutid die Veränderungen am Herzmuskel beeinflussen kann. Um die Fragestellungen zu klären, wurden verschiedene Untersuchungen in der Zellkultur durchgeführt. Skelettmuskel Myoblasten (undifferenzierte C2C12-Zellen), Skelettmuskel Myotuben (differenzierte C2C12-Zellen) und neonatale Rattenkardiomyozyten wurden unterschiedlichen Glukosekonzentrationen (5mM, 12mM, 25mM) für 24 oder 72 Stunden ausgesetzt. Die Herzmuskelzellen wurden zusätzlich in den Glukosekonzentrationen unter Zusatz von 12 mg/ml Liraglutid für 72h inkubiert. Die Expression der E3-Ligasen wurde mit qRT-PCR (MuRF-1 und MAFbx) und Western Blot (MuRF-1) quantifiziert. Die Poly-Ubiquitinylierung wurde mittels Western Blot bestimmt. Unter Verwendung des Cell Death Detection ELISA (Roche Diagnostics) wurde die Apoptoserate evaluiert. Die Inkubation von Skelett- und Herzmuskelzellen für 24 h mit hyperglykämischen Medium hatte kaum einen Einfluss auf die Expression von MuRF-1 und MAFbx sowie die Apoptoserate. Bei einer Inkubationszeit von 72 h konnten signifikante Erhöhungen bei 25mM Glukose für MuRF-1, MAFbx, der Poly-Ubiquitinylierung und der Apoptoserate in Skelett und Herzmuskelzellen festgestellt werden. Diese Anstiege konnten durch den Zusatz von Liraglutid beim Herzmuskel verhindert werden. Die Ergebnisse der vorliegenden Studie konnten zeigen, dass eine Hyperglykämie in der Zellkultur nach 72 h die Expression von zwei wichtigen Ubiquitin-E3-Ligasen sowie die Steigerung der Apoptoserate induzieren kann und dass dies durch Liraglutid am Herzmuskel verhindert werden kann. Diese Vorgänge können eventuell den Proteinverlust bzw. die Muskelatrophie beim Diabetes mellitus zum Teil erklären. Durch die positive Beeinflussung von Liraglutid an den Herzmuskelzellen könnte sich hier ein therapeutisches Potential zur Muskelatrophiebehandlung ergeben.

Obesity has grown to epidemic proportions, and in lack of efficient life-style and medical treatments, the bariatric surgeries are performed in rising numbers. The most common surgery is the Gastric Bypass (GBP) surgery, with the Biliopancreatic diversion with duodenal switch (DS) as an option for the most extreme cases with a BMI>50 kg/m2. In paper I 20 GBP-patients were examined during the first post-operative year regarding the natriuretic peptide, NT-ProBNP, which is secreted from the cardiac ventricles. Levels of NT-ProBNP quickly increased during the first post-surgery week, and later established itself on a higher level than pre-surgery. In paper II we report of 5 patient-cases after GBP-surgery with severe problems with postprandial hypoglycaemia that were successfully treated with GLP-1-analogs. The effect of treatment could be observed both symptomatically and in some cases using continuous glucose measuring systems (CGMS). In paper III three groups of subjects; 15 post-GBP patients, 15 post-DS, and 15 obese controls were examined for three days using CGMS during everyday life. The post-GBP group had high glucose variability as measured by MAGE and CONGA, whereas the post-DS group had low variability. Both post-operative groups exhibited significant time in hypoglycaemia, about 40 and 80 minutes per day <3.3mmol/l and 20 and 40 minutes < 2.8mmol/l, respectively, longer time for DS-group. Remarkably, only about 20% of these hypoglycaemic episodes were accompanied with symptoms. In Paper IV the hypoglycaemia counter regulatory system was investigated; 12 patients were examined before and after GBP-surgery with a stepped hypoglycaemic hyperinsulinemic clamp. The results show a downregulation of symptoms, counter regulatory hormones (glucagon, cortisol, epinephrine, norepinephrine, growth hormone), incretin hormones (GLP-1 and GIP), and sympathetic nervous response. In conclusion patients post bariatric surgery exhibit a downregulated counter regulatory response to hypoglycaemia, accompanied by frequent asymptomatic hypoglycaemic episodes in everyday life. Patients suffering from severe hypoglycaemic episodes can often be treated successfully with GLP-1-analogues.

The prevalence of childhood obesity and Type 2 Diabetes Mellitus (T2DM) has increased during recent decades. T2DM is accompanied with functional changes in the islets of Langerhans, which can be identified early in the pathogenesis. The aim of this thesis was to explore how metabolic changes caused by obesity early in life relate to islet function prior to overt T2DM. To address this, Uppsala Longitudinal Study of Childhood Obesity (ULSCO) was established (paper I). Initially, the association between palmitate and insulin secretion was investigated using a translational approach with obese and lean normoglycemic juveniles and isolated human islets (paper II). Secondly, dynamics of islet-hormones insulin and glucagon, and gut-hormones glucagon like-peptide 1 (GLP-1) and glicentin (paper III) and magnetic resonance imaging of pancreatic fat fraction (PFF) (paper IV) were studied in association to glucose tolerance and beta-cell function. Finally, a novel method of analysing shape features of oral glucose tolerance test (OGTT) curves was introduced and evaluated (paper V). Obese subjects had high prevalence of prediabetes and metabolic syndrome (MetS) (paper I). In obese pre-pubertal children with elevated palmitate levels, hyperinsulinemia was observed (paper II). In contrast, obese pubertal adolescents with similar palmitate levels showed moderate insulin levels during OGTT with delayed first phase insulin response. To explore mechanisms for these variations, isolated human islets were exposed to palmitate for different time periods in vitro. After 2 days accentuated insulin response was observed. Impaired beta-cell function and apoptosis were evident after 7 days, however. Hyperglucagonemia and disturbed GLP-1 and glicentin levels were associated with obesity and glycaemic status, with fasting glicentin being predictive of prediabetes (paper III). Furthermore, PFF was increased in obese subjects and associated to MetS and visceral adipose tissue, but not to beta-cell function (paper IV). OGTT curves were converted into geometric centres, centroids, which correlated with differences in glucose tolerance (paper V). In conclusion, the islet function in obese children was associated with elevated levels of palmitate, but not pancreatic fat. Fasting palmitate and glicentin levels, as well as centroid analyses of OGTT curves, could potentially identify obese children at risk of prediabetes and subsequent T2DM.

Made available in DSpace on 2016-06-02T20:37:48Z (GMT). No. of bitstreams: 1 5175.pdf: 1937380 bytes, checksum: c74ac55f5059fbe2d78f9dabe02dc226 (MD5) Previous issue date: 2013-06-07 / The tests and/or study laboratories have sought the implementation of Quality Management Systems (QMS) in order to obtain a differential, which is the greater reliability and acceptance of their results. The two main systems adopted in laboratories are the Good Laboratory Practices (GLP), which guidelines are defined in NIT-DICLA-035, and ISO/IEC 17025, according to NBR ISO/IEC 17025. Comparatively, the guidelines of both standards, in respect to the experimental stage of laboratory, have the same goal, to guarantee the quality of the results. The systems differ in that the ISO/IEC 17025 operates in the laboratory routine activities as a whole, in contrast GLP focus is to serve as a base to a study that is accomplished. An analysis of the elements of the standards shows that there is substantial overlap in both of the requirements, which makes it possible to maintain a single and integrated system. Thus, this paper aims to analyze critically the implementation and maturation process of an integrated QMS according to GLP and ISO/IEC 17025 implemented in a Laboratory that performs tests of pesticides and veterinary products residues using chromatography. The system implementation was done in stages, and the activities performed in each one were evaluated. The first step consisted in defining the quality assurance team. Next it was defined the work scope, followed by the steps: analysis of the similarities between the standards and structuring of a system; adaptation and elaboration of documentation to comply with the integrated system, with emphasis on the methods validation and uncertainty estimation procedures; training; audits; critical analysis meeting by management and finally organize the documentation for submission to INMETRO. During the implementation process it was conducted a survey among collaborators about the difficulties and benefits of the process. In addition, it was evaluated the time required to implement the system, the period to get the formal recognition and audit data, such as noncompliance highlighted and corrective actions taken. These data were compared with those obtained in Brazil and the USA laboratories. / Os laboratórios de estudo e/ou ensaios tem buscado a implementação de Sistemas de Gestão da Qualidade (SGQ) como forma de obter um diferencial, que reside na maior confiabilidade e aceitação de seus resultados. Os dois principais sistemas adotados em laboratórios são as Boas Práticas de Laboratório (BPL), cujas diretrizes são definidas na NIT-DICLA-035, e a ISO/IEC 17025, conforme NBR ISO/IEC 17025. Comparativamente, as diretrizes de ambas as normas, no que se refere à etapa experimental do laboratório, possuem o mesmo objetivo, a garantia da qualidade dos resultados obtidos. Os sistemas se diferem na medida em que a ISO/IEC 17025 atua nas atividades rotineiras do laboratório como um todo, já o foco das BPL é dar embasamento a um estudo realizado. Uma análise dos elementos das normas evidencia que há grande sobreposição dos requisitos exigidos em ambas, o que torna possível manter um sistema único e integrado. Assim, o presente trabalho tem como objetivo analisar criticamente o processo de implementação e amadurecimento de um SGQ integrado conforme as BPL e a ISO/IEC 17025 implementado em um Laboratório que realiza análises de resíduos de agrotóxicos e produtos veterinários por cromatografia. A implementação do sistema foi realizada em etapas, sendo que as atividades realizadas em cada uma delas foram avaliadas. A primeira etapa consistiu na definição da equipe de garantia da qualidade. Na sequência foi definido o escopo de trabalho, seguida das etapas: análise das similaridades entre as normas e estruturação de um sistema; adaptação e elaboração de documentação para atendimento ao sistema integrado, com destaque para os procedimentos de validação de métodos e estimativa da incerteza; treinamentos; auditorias; reunião de análise crítica pela direção e por fim a montagem da documentação para envio ao INMETRO. Durante o processo de implementação foi feita uma pesquisa entre os colaboradores a respeito das dificuldades e benefícios do processo. Além disso, foi avaliado o período necessário para implementar o sistema, o período para obter o reconhecimento formal e dados das avaliações, tais como as não conformidades evidenciadas e as ações corretivas tomadas. Esses dados foram comparados com os de laboratórios no Brasil e nos EUA.

The aim of this research was the development of mass spectrometry-based methodologies for the high-throughput quantitation of peptides in biological matrices. Glucagon and GLP-1, which are of interest as biomarkers and in the development of therapeutics, were chosen as model peptides. Immunoassays that are traditionally used to quantify these often perform poorly; therefore, necessitating the development of alternative methodologies. Application of mass spectrometry-based methodologies to these analytes has, however, been limited, primarily due to sensitivity challenges, but also due to analytical challenges associated with their endogenous nature and instability in biological matrices. Chapter 2 describes the development and qualification of the first liquid-chromatography coupled tandem mass spectrometry (LC-MS/MS) method for the quantitation of endogenous glucagon from human plasma. A novel 2D extraction procedure was developed to ensure robustness and sensitivity, whilst a novel surrogate matrix quantitation strategy took into account the endogenous nature of the analyte. A lower limit of quantitation (LLOQ) of 25 pg/mL was qualified, which was a considerable improvement over that previously reported in the literature (250 pg/mL) for a LC-MS/MS method. Clinical samples were cross-validated against a conventional radioimmunoassay (RIA), and similar pharmacokinetic (PK) profiles resulted, demonstrating that the methods were complementary. In Chapter 2 glucagon instability in biological matrix was noted. To characterise this further, in Chapter 3 in vitro glucagon metabolites were identified using high-resolution mass spectrometry (HRMS). Metabolites observed by others (glucagon19-29, glucagon3 29 and [pGlu]3glucagon3 29) in alternative matrices were identified, alongside novel metabolites (glucagon20-29 and glucagon21-29). Cross-interference of these metabolites in immunoassays may help to explain their poor performance, whilst knowledge of metabolism may also aid the development of future stabilisation strategies. The method developed in Chapter 2 was refined in Chapter 4 to improve sensitivity, robustness and throughput, and to add GLP-1 as a secondary analyte. The sensitivity achieved (glucagon: 15 pg/mL LLOQ, GLP-1: 25 pg/mL LLOQ) is the highest reported for both peptides for an extraction avoiding immunoenrichment. Specificity of endogenous glucagon quantitation was assured using a novel approach with a supercharging mobile phase additive to access a sensitive qualifier transition. A cross-validation against established immunoassays using physiological study samples demonstrated some similarities between the methods. Differences between the immunoassay results exemplified the need to develop alternative methodologies. The resulting LC-MS/MS method is considered a viable alternative to immunoassays, for the quantitation of endogenous glucagon, dosed glucagon and/or dosed GLP-1 in human plasma.

Made available in DSpace on 2009-11-18T19:01:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2004 / A indústria brasileira de gás liquefeito de petróleo, GLP, tem sido alvo freqüente, nos últimos anos, da atenção de órgãos reguladores, mídia e público em geral. Muitas têm sido as especulações a respeito de seu padrão de conduta publicados na imprensa, variando desde práticas predatórias até a formação disciplinada de cartel. A presente pesquisa, baseada em depoimentos de executivos da indústria e no acesso a dados das firmas dessa indústria tomado públicos - dados financeiros e operacionais - explora qual seja, afinal, o padrão de competição desse oligopólio. O estudo parte da Teoria da Estrutura de Capital aplicada para decisões de competição e mercado - preços e quantidades a ofertar para constatar que, dadas suas escolhas de estrutura de capital, as firmas esta indústria específica simplesmente adotam um padrão tradicional de arrefecimento da competição a partir de sua dívida. A dívida, nesta indústria, é utilizada para a criação de barreiras de entrada. Ainda assim, casos de acirramento da competição e de concentração de mercado foram encontrados, indicando que não haja uma disciplina formal de conduta por parte de suas firmas as quais devem, eventualmente, encontrar dificuldades - ou falta de interesse - na convivência em um mercado em fase de transformação. / The Brazilian industry of liquefied petroleum gas, LPG, has been calling, in the late years, the attention of the media, regulatory agencies and the general publico Many speculations have emerged, in the press, over its pattems of behaviour, which have been ranging from predatory practices up to a formal cartel operating under collusion. The present research, based in statements from industry executives as well as in data publicly disclosed by those firms - financiaI and operational ones - explores what their competitive behaviour would be, after alI. The study starts from the Theory of Capital Structure applied to product market competition decisions - output and price decisions - to realize that, given their capital structure choices, the firms within that particular industry simply adopt a classical behaviour of sofier competition. Debt, in this industry, has the role of an entry barrier. Even though, cases of tougher competition have been found, indicating the lack of a formal discipline among its members which ones, eventually, may find it very difficult - or of no interest - to coexist together in a fast changing industrial environment.

Es ist bekannt, dass eine diabetische Stoffwechsellage über einen gesteigerten Proteinabbau zu einer Muskelatrophie führen kann. Ein zentrales System beim Abbau von Muskelproteinen ist hierbei das Ubiquitin-Proteasom-System mit seinen zwei spezifischen E3-Ligasen MuRF-1 und MAFbx. Ziel dieser Arbeit war es, den Einfluss einer Hyperglykämie auf die Expression von Proteinen des Ubiquitin-Proteasom-Systems im Skelett- und Herzmuskel zeit- und konzentrationsabhängig zu untersuchen. Insbesondere stand die Expression der E3-Ligasen MuRF-1 und MAFbx sowie die daraus folgende Auswirkung auf die Protein-Ubiquitinylierung im Fokus der Untersuchungen. Weiterhin sollte der Einfluss der Hyperglykämie auf die Apoptoserate von Skelett- und Herzmuskelzellen analysiert werden. Seit Kurzem stehen die GLP-1-Analoga als neue Antidiabetika für die Therapie des Diabetes mellitus Type II zur Verfügung. Da in Studien bereits positive Effekte der GLP-1-Analoga am Herzmuskel festgestellt wurden, sollte in der vorliegenden Studie geprüft werden, ob das GLP-1-Analogon Liraglutid die Veränderungen am Herzmuskel beeinflussen kann. Um die Fragestellungen zu klären, wurden verschiedene Untersuchungen in der Zellkultur durchgeführt. Skelettmuskel Myoblasten (undifferenzierte C2C12-Zellen), Skelettmuskel Myotuben (differenzierte C2C12-Zellen) und neonatale Rattenkardiomyozyten wurden unterschiedlichen Glukosekonzentrationen (5mM, 12mM, 25mM) für 24 oder 72 Stunden ausgesetzt. Die Herzmuskelzellen wurden zusätzlich in den Glukosekonzentrationen unter Zusatz von 12 mg/ml Liraglutid für 72h inkubiert. Die Expression der E3-Ligasen wurde mit qRT-PCR (MuRF-1 und MAFbx) und Western Blot (MuRF-1) quantifiziert. Die Poly-Ubiquitinylierung wurde mittels Western Blot bestimmt. Unter Verwendung des Cell Death Detection ELISA (Roche Diagnostics) wurde die Apoptoserate evaluiert. Die Inkubation von Skelett- und Herzmuskelzellen für 24 h mit hyperglykämischen Medium hatte kaum einen Einfluss auf die Expression von MuRF-1 und MAFbx sowie die Apoptoserate. Bei einer Inkubationszeit von 72 h konnten signifikante Erhöhungen bei 25mM Glukose für MuRF-1, MAFbx, der Poly-Ubiquitinylierung und der Apoptoserate in Skelett und Herzmuskelzellen festgestellt werden. Diese Anstiege konnten durch den Zusatz von Liraglutid beim Herzmuskel verhindert werden. Die Ergebnisse der vorliegenden Studie konnten zeigen, dass eine Hyperglykämie in der Zellkultur nach 72 h die Expression von zwei wichtigen Ubiquitin-E3-Ligasen sowie die Steigerung der Apoptoserate induzieren kann und dass dies durch Liraglutid am Herzmuskel verhindert werden kann. Diese Vorgänge können eventuell den Proteinverlust bzw. die Muskelatrophie beim Diabetes mellitus zum Teil erklären. Durch die positive Beeinflussung von Liraglutid an den Herzmuskelzellen könnte sich hier ein therapeutisches Potential zur Muskelatrophiebehandlung ergeben.

El presente trabajo muestra la situación actual de la demanda de Gas Liquado de Petroleo (GLP) en el mercado peruano con respecto al parque automotor durante los últimos 6 años. El objetivo general es predecir la demanda de GLP para el segundo semestre del año 2021, a través de las variables más relevantes a fin de conocer si la producción local más la importación de este tipo de combustible (GLP) será la suficiente para cubrir la demanda del sector automotriz. La metodología utilizada por el equipo de ciencia de datos es Cross Industry Standard Process for Data Mining (CRISP-DM), la cual consiste en seguir una serie de diez etapas, en cada una de ellas se ira descubriendo y analizando las variables que serán relevantes para la elaboración del modelo deseado. El modelo seleccionado por el equipo de ciencia de datos es el modelo de aprendizaje predictivo ya que este agrupa varias técnicas estadísticas de modelización, lo cual incluye algoritmos de aprendizaje automático. Posteriormente las Herramientas que se utilizarán para un mejor Análisis y entendimiento de la problemática serán Power BI, KNime y Python. / This paper shows the current situation of Liquefied Petroleum Gas (LPG) demand in the Peruvian market with respect to the vehicle fleet during the last 6 years. The general objective is to predict the LPG demand for the second semester of the year 2021, through the most relevant variables to know if the local production plus the import of this type of fuel (LPG) will be enough to cover the demand of the automotive sector. The methodology used by the data science team is Cross Industry Standard Process for Data Mining (CRISP-DM), which consists of following a series of ten stages, in each of which the variables that will be relevant for the elaboration of the desired model will be discovered and analyzed. The model selected by the data science team is the predictive learning model because it groups several statistical modeling techniques, including machine learning algorithms. Subsequently, the tools to be used for a better analysis and understanding of the problem will be Power BI, KNime and Python. / Trabajo de investigación

Das Darmhormon Glucagon-like peptide-1 (GLP-1) ist ein vielversprechendes Therapeutikum in der Behandlung des Typ 2-Diabetes. Allerdings liegen nur wenige Daten über die molekulare Wirkung des Hormons vor. Daher wurden die intrazellulären Effekte, die GLP-1-induzierten Signaltransduktion und Genexpression am Beispiel der pankreatischen ß-Zellinie INS-1 untersucht. In den INS-1-Zellen zeigte sich nach einer GLP-1-Stimulation eine zeit-, dosis- und glukose-abhängige Phosphorylierung von ERK1/2 bzw. der Aktivierung der Transkriptionsfaktoren Elk-1 und CREB. Während GLP-1 die beiden Signalmoleküle bereits nach wenigen Minuten transient aktivierte, führte Glukose zu einer verzögerten, aber anhaltenden Aktivierung. Beide Stimulanzien gemeinsam bewirkten eine synergistische Aktivierung von ERK1/2 und CREB. Sowohl am Mechanismus der glukose- als auch GLP-1-induzierten Aktivierung sind Ca2[plus]-regulierte Signalprozesse in antagonistischer Weise involviert. So führte die Inhibition von CaM-Kinasen und der intrazellulären Ca2[plus]-Erhöhung zu einer Reduktion der GLP-1- und glukosestimulierten ERK1/2-Phosphorylierung, die induzierte Aktivierung von CREB wurde leicht verstärkt. Die Inhibition der PKA und MEK ließen die Rückschlüsse zu, daß die Aktivie-rung der MAPK-Kaskade teilweise durch die PKA vermittelt wird und eine Wechselwirkung zwischen den Kaskaden existiert. Insbesondere die Induktion der frühen Gene junB, c-fos, nur-77 und zif268, die mit der GLP-1-induzierten ERK1/2- und CREB-Phosphorylierung korrelierten, belegten, daß GLP-1 einen Teil seiner Wirkung über diese Signalwege vermittelt. Die Glukosewirkung hinsichtlich der IEG-Induktion war gering. Beide Stimuli gemeinsam führten analog zum synergistischen Effekt auf signaltransduktorischer Ebene zu einer gesteigerten und verlängerten IEG-Expression, die somit die physiologische Bedeutung von GLP-1 als Glukosekompetenzfaktor unterstrichen.