Um estudo historiográfico sobre a psicanálise com crianças no contexto universitário

Pós-graduação em Psicologia - FCLAS

Clinical Effects of Scalp Electrical Acupuncture in Stroke: A Sham-Controlled Randomized Clinical Trial

Objectives: The majority of individuals who survive a stroke are disabled because of persisting neurological impairments. The objective of this study was to evaluate the efficacy of subcutaneous electrical stimulation of the scalp in spontaneous functional recovery of patients with chronic ischemic stroke, by evaluating clinical, neurological, and functional findings. Subjects and methods: Sixty-two (62) subjects who were at least 18 months postdiagnosis of ischemic stroke were randomized to receive 10 sessions of placebo or active low-frequency electrical stimulation (2/100 Hz) using subcutaneous acupuncture needles over the scalp. Functional and neurological evaluations were indexed by the Barthel, Rankin, and National Institutes of Health Stroke Scale (NIHSS). Results: Results show that there was a significant difference in functional improvement between the sham and active group as indexed by NIHSS scale. The active group had a larger functional improvement after 10 sessions of scalp electrical acupuncture. The other two functional scales (Rankin and Barthel) failed to show significant differences between the two treatment groups. Conclusions: These results support further testing of scalp electrical acupuncture for the treatment of stroke as well further mechanistic studies to understand mechanisms associated with the observed improvement. Further studies need to consider longer follow-up assessments to investigate potential functional changes associated with electrical acupuncture.

An integrated approach to evaluate emerging contaminants in drinking water

The chemical stock of emerging contaminants in Brazilian drinking water is of great interest due to the poor water quality at surface water intakes. In addition, little is known about the effect of some contaminants, such as endocrine disrupting chemicals (EDCs), which may be present in both raw and treated water. The aim of this work was to evaluate selected emerging contaminants in Brazilian waters using both chemical and biological analyses. Sampling sites were established in different municipalities based upon raw water quality data. Estrone, 17 beta-estradiol, estriol, 17 alpha-ethinylestradiol, bisphenol A, 4-n-octylphenol and 4-n-nonylphenol were determined in the samples by liquid chromatography-tandem mass spectrometry. A yeast assay using a Saccharomyces cerevisiae bioluminescent bioreporter was used to evaluate the estrogenic activity of the water samples. The first integrated results revealed similarities between the two individual approaches, since higher values for the bioassay were accompanied by significant concentrations of some selected compounds in surface water samples. No estrogenicity was observed for drinking water samples. Our results also indicate that the usual paradigm of evaluating water quality by measuring selected EDCs in a given water sample via chemical analysis, needs to be reviewed since the observed estrogenicity of a water sample is now a better guiding parameter to the selection of samples and substances to be chemically investigated in further analysis. So far, the data bank produced in this work, i.e., the comparison between chemical burden and observed estrogenicity, is not yet sufficiently robust to fully guide this decision. (C) 2011 Elsevier B.V. All rights reserved.

Angiotensin II Facilitates Breast Cancer Cell Migration and Metastasis

Breast cancer metastasis is a leading cause of death by malignancy in women worldwide. Efforts are being made to further characterize the rate-limiting steps of cancer metastasis, i.e. extravasation of circulating tumor cells and colonization of secondary organs. In this study, we investigated whether angiotensin II, a major vasoactive peptide both produced locally and released in the bloodstream, may trigger activating signals that contribute to cancer cell extravasation and metastasis. We used an experimental in vivo model of cancer metastasis in which bioluminescent breast tumor cells (D3H2LN) were injected intra-cardiacally into nude mice in order to recapitulate the late and essential steps of metastatic dissemination. Real-time intravital imaging studies revealed that angiotensin II accelerates the formation of metastatic foci at secondary sites. Pre-treatment of cancer cells with the peptide increases the number of mice with metastases, as well as the number and size of metastases per mouse. In vitro, angiotensin II contributes to each sequential step of cancer metastasis by promoting cancer cell adhesion to endothelial cells, trans-endothelial migration and tumor cell migration across extracellular matrix. At the molecular level, a total of 102 genes differentially expressed following angiotensin II pretreatment were identified by comparative DNA microarray. Angiotensin II regulates two groups of connected genes related to its precursor angiotensinogen. Among those, up-regulated MMP2/MMP9 and ICAM1 stand at the crossroad of a network of genes involved in cell adhesion, migration and invasion. Our data suggest that targeting angiotensin II production or action may represent a valuable therapeutic option to prevent metastatic progression of invasive breast tumors.

No evidence of selenosis from a selenium-rich diet in the Brazilian Amazon

Selenium (Se) is an essential element and a well-known anti-oxidant. In the Lower Tapajos River region of the Brazilian Amazon, biomarkers of Se range from normal to very high. The local traditional diet includes important Se sources such as Brazil nuts, chicken, game meat and certain fish species. Some studies have reported alterations in keratin structure, gastrointestinal problems and paresthesia in populations with high Se intake. The objective of the present study was to evaluate cutaneous and garlic odor of the breath signs and sentinel symptoms of Se toxicity (selenosis) in relation to Se status in communities along the Tapajos River. Participants (N = 448), aged 15-87 years, were recruited from 12 communities. Se concentrations were measured in blood (B-Se) and plasma (P-Se) by ICP-MS. A nurse performed an examination of the hair, nails, skin and breath for signs of Se toxicity. Interview-administered questionnaires were used to collect information on socio-demographics, medical history and possible symptoms of Se toxicity. In this population, the median levels of B-Se and P-Se were 228.4 mu g/L (range 103.3-1500.2 mu g/L) and 134.8 mu g/L (range 53.6-913.2 mu g/L) respectively. Although B-Se and P-Se surpassed concentrations considered toxic (B-Se: 1000 mu g/L (U.S. EPA, 2002)), no dermal or breath signs or symptoms of Se toxicity were associated with the biomarkers of Se status. In the present study population, where Se intake is mostly from traditional diet, there is no evidence of selenosis. These findings support the need to re-assess Se toxicity considering factors such as the chemical form of Se exposure, route of exposure (inhaled versus ingested), co-exposures to toxic elements such as mercury. Considering the current food transition towards a western diet in the Amazon, further studies should address the possible association between high Se status and cardiometabolic health in this study population. (C) 2011 Elsevier Ltd. All rights reserved.

THE SLOAN DIGITAL SKY SURVEY CO-ADD: A GALAXY PHOTOMETRIC REDSHIFT CATALOG

We present and describe a catalog of galaxy photometric redshifts (photo-z) for the Sloan Digital Sky Survey (SDSS) Co-add Data. We use the artificial neural network (ANN) technique to calculate the photo-z and the nearest neighbor error method to estimate photo-z errors for similar to 13 million objects classified as galaxies in the co-add with r < 24.5. The photo-z and photo-z error estimators are trained and validated on a sample of similar to 83,000 galaxies that have SDSS photometry and spectroscopic redshifts measured by the SDSS Data Release 7 (DR7), the Canadian Network for Observational Cosmology Field Galaxy Survey, the Deep Extragalactic Evolutionary Probe Data Release 3, the VIsible imaging Multi-Object Spectrograph-Very Large Telescope Deep Survey, and the WiggleZ Dark Energy Survey. For the best ANN methods we have tried, we find that 68% of the galaxies in the validation set have a photo-z error smaller than sigma(68) = 0.031. After presenting our results and quality tests, we provide a short guide for users accessing the public data.

Mutation Spectrum in the Large GTPase Dynamin 2, and Genotype-Phenotype Correlation in Autosomal Dominant Centronuclear Myopathy

Centronuclear myopathy (CNM) is a genetically heterogeneous disorder associated with general skeletal muscle weakness, type I fiber predominance and atrophy, and abnormally centralized nuclei. Autosomal dominant CNM is due to mutations in the large GTPase dynamin 2 (DNM2), a mechanochemical enzyme regulating cytoskeleton and membrane trafficking in cells. To date, 40 families with CNM-related DNM2 mutations have been described, and here we report 60 additional families encompassing a broad genotypic and phenotypic spectrum. In total, 18 different mutations are reported in 100 families and our cohort harbors nine known and four new mutations, including the first splice-site mutation. Genotype-phenotype correlation hypotheses are drawn from the published and new data, and allow an efficient screening strategy for molecular diagnosis. In addition to CNM, dissimilar DNM2 mutations are associated with Charcot-Marie-Tooth (CMT) peripheral neuropathy (CMTD1B and CMT2M), suggesting a tissue-specific impact of the mutations. In this study, we discuss the possible clinical overlap of CNM and CMT, and the biological significance of the respective mutations based on the known functions of dynamin 2 and its protein structure. Defects in membrane trafficking due to DNM2 mutations potentially represent a common pathological mechanism in CNM and CMT. Hum Mutat 33: 949-959, 2012. (C) 2012 Wiley Periodicals, Inc.

Regional differences in clinical care among patients with type 1 diabetes in Brazil: Brazilian Type 1 Diabetes Study Group

Background To determine the characteristics of clinical care offered to type 1 diabetic patients across the four distinct regions of Brazil, with geographic and contrasting socioeconomic differences. Glycemic control, prevalence of cardiovascular risk factors, screening for chronic complications and the frequency that the recommended treatment goals were met using the American Diabetes Association guidelines were evaluated. Methods This was a cross-sectional, multicenter study conducted from December 2008 to December 2010 in 28 secondary and tertiary care public clinics in 20 Brazilian cities in north/northeast, mid-west, southeast and south regions. The data were obtained from 3,591 patients (56.0% females and 57.1% Caucasians) aged 21.2 ± 11.7 years with a disease duration of 9.6 ± 8.1 years (<1 to 50 years). Results Overall, 18.4% patients had HbA1c levels <7.0%, and 47.5% patients had HbA1c levels ≥ 9%. HbA1c levels were associated with lower economic status, female gender, age and the daily frequency of self-blood glucose monitoring (SBGM) but not with insulin regimen and geographic region. Hypertension was more frequent in the mid-west (32%) and north/northeast (25%) than in the southeast (19%) and south (17%) regions (p<0.001). More patients from the southeast region achieved LDL cholesterol goals and were treated with statins (p<0.001). Fewer patients from the north/northeast and mid-west regions were screened for retinopathy and nephropathy, compared with patients from the south and southeast. Patients from the south/southeast regions had more intensive insulin regimens than patients from the north/northeast and mid-west regions (p<0.001). The most common insulin therapy combination was intermediate-acting with regular human insulin, mainly in the north/northeast region (p<0.001). The combination of insulin glargine with lispro and glulisine was more frequently used in the mid-west region (p<0.001). Patients from the north/northeast region were younger, non-Caucasian, from lower economic status, used less continuous subcutaneous insulin infusion, performed less SBGM and were less overweight/obese (p<0.001). Conclusions A majority of patients, mainly in the north/northeast and mid-west regions, did not meet metabolic control goals and were not screened for diabetes-related chronic complications. These results should guide governmental health policy decisions, specific to each geographic region, to improve diabetes care and decrease the negative impact diabetes has on the public health system.

New insights in osteogenic differentiation revealed by mass spectrometric assessment of phosphorylated substrates in murine skin mesenchymal cells

Abstract Background Bone fractures and loss represent significant costs for the public health system and often affect the patients quality of life, therefore, understanding the molecular basis for bone regeneration is essential. Cytokines, such as IL-6, IL-10 and TNFα, secreted by inflammatory cells at the lesion site, at the very beginning of the repair process, act as chemotactic factors for mesenchymal stem cells, which proliferate and differentiate into osteoblasts through the autocrine and paracrine action of bone morphogenetic proteins (BMPs), mainly BMP-2. Although it is known that BMP-2 binds to ActRI/BMPR and activates the SMAD 1/5/8 downstream effectors, little is known about the intracellular mechanisms participating in osteoblastic differentiation. We assessed differences in the phosphorylation status of different cellular proteins upon BMP-2 osteogenic induction of isolated murine skin mesenchymal stem cells using Triplex Stable Isotope Dimethyl Labeling coupled with LC/MS. Results From 150 μg of starting material, 2,264 proteins were identified and quantified at five different time points, 235 of which are differentially phosphorylated. Kinase motif analysis showed that several substrates display phosphorylation sites for Casein Kinase, p38, CDK and JNK. Gene ontology analysis showed an increase in biological processes related with signaling and differentiation at early time points after BMP2 induction. Moreover, proteins involved in cytoskeleton rearrangement, Wnt and Ras pathways were found to be differentially phosphorylated during all timepoints studied. Conclusions Taken together, these data, allow new insights on the intracellular substrates which are phosphorylated early on during differentiation to BMP2-driven osteoblastic differentiation of skin-derived mesenchymal stem cells.

Tolkning av rekvisitet ”Annat socialt nedbrytande beteende” i § 3 LVU

Syfte med uppsatsen är att ge en bättre förståelse av vad som menas med begreppet ”annat socialt nedbrytande beteende” i § 3 LVU och visa eventuella skillnader i hur lagstiftningen, domstolar och socialtjänsten använder detta begrepp. Vi har undersökt hur begreppsdefinitionen har utvecklats genom lagstiftningsprocessen och hur den återspeglas i förarbeten, juridisk litteratur, prejudikat och praxis. Uppsatsen är skriven utifrån en juridisk metod. Vi genomförde åtta semistrukturerade intervjuer med socialsekreterare i Hälsingland och en länsrättsdomare för att ta reda på hur dessa använder begreppet. Vi har gjort en sammanställning av olika problembeteenden som ungdomar kan uppvisa och diskuterat om dessa är socialt nedbrytande beteende i lagens mening. I lagstiftningen är definitionen av rekvisitet diffus och generell, detta innebär problem för socialsekreterare att använda rekvisitet. Socialsekreterarna är inte vana och tillräckligt utbildade att använda begreppet vilket kan leda till en mindre rättssäker prövning för den enskilde.

High-resolution stratigraphy of Central and Southern Adriatic Quaternary deposits of sub-Milankovian climate change on Mediterranean circulation

This volume is a collection of the work done in a three years-lasting PhD, focused in the analysis of Central and Southern Adriatic marine sediments, deriving from the collection of a borehole and many cores, achieved thanks to the good seismic-stratigraphic knowledge of the study area. The work was made out within European projects EC-EURODELTA (coordinated by Fabio Trincardi, ISMAR-CNR), EC-EUROSTRATAFORM (coordinated by Phil P. E. Weaver, NOC, UK), and PROMESS1 (coordinated by Serge Bernè, IFREMER, France). The analysed sedimentary successions presented highly expanded stratigraphic intervals, particularly for the last 400 kyr, 60 kyr and 6 kyr BP. These three different time-intervals resulted in a tri-partition of the PhD thesis. The study consisted of the analysis of planktic and benthic foraminifers’ assemblages (more than 560 samples analysed), as well as in preparing the material for oxygen and carbon stable isotope analyses, and interpreting and discussing the obtained dataset. The chronologic framework of the last 400 kyr was achieved for borehole PRAD1-2 (within the work-package WP6 of PROMESS1 project), collected in 186.5 m water depth. The proposed chronology derives from a multi-disciplinary approach, consisting of the integration of numerous and independent proxies, some of which analysed by other specialists within the project. The final framework based on: micropaleontology (calcareous nannofossils and foraminifers’ bioevents), climatic cyclicity (foraminifers’ assemblages), geochemistry (oxygen stable isotope, made out on planktic and benthic records), paleomagnetism, radiometric ages (14C AMS), teprhochronology, identification of sapropel-equivalent levels (Se). It’s worth to note the good consistency between the oxygen stable isotope curve obtained for borehole PRAD1-2 and other deeper Mediterranean records. The studied proxies allowed the recognition of all the isotopic intervals from MIS10 to MIS1 in PRAD1-2 record, and the base of the borehole has been ascribed to the early MIS11. Glacial and interglacial intervals identified in the Central Adriatic record have been analysed in detail for the paleo-environmental reconstruction, as well. For instance, glacial stages MIS6, MIS8 and MIS10 present peculiar foraminifers’ assemblages, composed by benthic species typical of polar regions and no longer living in the Central Adriatic nowadays. Moreover, a deepening trend in the paleo-bathymetry during glacial intervals was observed, from MIS10 (inner-shelf environment) to MIS4 (mid-shelf environment).Ten sapropel-equivalent levels have been recognised in PRAD1-2 Central Adriatic record. They showed different planktic foraminifers’ assemblages, which allowed the first distinction of events occurred during warm-climate (Se5, Se7), cold-climate (Se4, Se6 and Se8) and temperate-intermediate-climate (Se1, Se3, Se9, Se’, Se10) conditions, consistently with literature. Cold-climate sapropel equivalents are characterised by the absence of an oligotrophic phase, whereas warm-temeprate-climate sapropel equivalents present both the oligotrophic and the eutrophic phases (except for Se1). Sea floor conditions vary, according to benthic foraminifers’ assemblages, from relatively well oxygenated (Se1, Se3), to dysoxic (Se9, Se’, Se10), to highly dysoxic (Se4, Se6, Se8) to events during which benthic foraminifers are absent (Se5, Se7). These two latter levels are also characterised by the lamination of the sediment, feature never observed in literature in such shallow records. The enhanced stratification of the water column during the events Se8, Se7, Se6, Se5, Se4, and the concurring strong dilution of shallow water, pointed out by the isotope record, lead to the hypothesis of a period of intense precipitation in the Central Adriatic region, possibly due to a northward shift of the African Monsoon. Finally, the expression of Central Adriatic PRAD1-2 Se5 equivalent was compared with the same event, as registered in other Eastern Mediterranean areas. The sequence of substantially the same planktic foraminifers’ bioevents has been consistently recognised, indicating a similar evolution of the water column all over the Eastern Mediterranean; yet, the synchronism of these events cannot be demonstrated. A high resolution analysis of late Holocene (last 6000 years BP) climate change was carried out for the Adriatic area, through the recognition of planktic and benthic foraminifers’ bioevents. In particular, peaks of planktic Globigerinoides sacculifer (four during the last 5500 years BP in the most expanded core) have been interpreted, based on the ecological requirements of this species, as warm-climate, arid intervals, correspondent to periods of relative climatic optimum, such as, for instance, the Medieval Warm Period, the Roman Age, the Late Bronze Age and the Copper Age. Consequently, the minima in the abundance of this biomarker could correspond to relatively cooler and more rainy periods. These conclusions are in good agreement with the isotopic and the pollen data. The Last Occurrence (LO) of G. sacculifer has been dated in this work at an average age of 550 years BP, and it is the best bioevent approximating the base of the Little Ice Age in the Adriatic. Recent literature reports the same bioevent in the Levantine Basin, showing a rather consistent age. Therefore, the LO of G. sacculifer has the potential to be extended to all the Eastern Mediterranean. Within the Little Ice Age, benthic foraminifer V. complanata shows two distinct peaks in the shallower Adriatic cores analysed, collected hundred kilometres apart, inside the mud belt environment. Based on the ecological requirements of this species, these two peaks have been interpreted as the more intense (cold and rainy) oscillations inside the LIA. The chronologic framework of the analysed cores is robust, being based on several range-finding 14C AMS ages, on estimates of the secular variation of the magnetic field, on geochemical estimates of the activity depth of 210Pb short-lived radionuclide (for the core-top ages), and is in good agreement with tephrochronologic, pollen and foraminiferal data. The intra-holocenic climate oscillations find out in the Adriatic have been compared with those pointed out in literature from other records of the Northern Hemisphere, and the chronologic constraint seems quite good. Finally, the sedimentary successions analysed allowed the review and the update of the foraminifers’ ecobiostratigraphy available from literature for the Adriatic region, thanks to the achievement of 16 ecobiozones for the last 60 kyr BP. Some bioevents are restricted to the Central Adriatic (for instance the LO of benthic Hyalinea balthica , approximating the MIS3/MIS2 boundary), others occur all over the Adriatic basin (for instance the LO of planktic Globorotalia inflata during MIS3, individuating Dansgaard-Oeschger cycle 8 (Denekamp)).

Einfluss des Aktin-Zytoskeletts und intrazellulärer Signalkaskaden auf die postsynaptische Lokalisation von Gephyrin

Neuronen haben für die Informationsübertragung untereinander spezielle Strukturen entwickelt, welche als Synapsen bezeichnet werden. Um eine schnelle und präzise synaptische Signalübertragung zu gewährleisten, ist eine hohe Konzentration von Neurotransmitter-regulierten Ionenkanälen in der postsynaptischen Plasmamembran notwendig. Die spezifische Verankerung der Rezeptoren wird durch intrazelluläre Proteine der Postsynapse vermittelt. Das periphere Membranprotein Gephyrin spielt eine essentielle Rolle in der synaptischen Lokalisation von Glyzin- und GABAA-Rezeptoren an inhibitorischen Synapsen. Um das postsynaptische Netzwerk zu stabilisieren, ist eine Interaktion von Gephyrin mit Proteinen der Mikrofilamente und der Mikrotubuli nötig. In der vorliegenden Arbeit sollte analysiert werden, wie Gephyrin mit dem Aktin-Zytoskelett interagiert, und ob die Größe und Stabilität neuronaler Gephyrincluster durch das Aktin-Zytoskelett reguliert wird. Dies wurde mittels Expression von GFP-Gephyrin-Konstrukten in HEK293T-Zellen und Aktin-depolymerisierende Alkaloidbehandlung von hippokampalen Primärkulturzellen untersucht. Der Einfluss unterschiedlicher Signalkaskaden auf die Lokalisation und Funktionalität von GABAA- oder Glyzin-Rezeptoren wurde bereits intensiv untersucht, jedoch für Rezeptor-assoziierte Proteine wie Gephyrin existierten nur wenig relevante Daten. Ein weiteres Ziel war daher, durch pharmakologische Beeinflussung von Schlüsselenzymen in hippokampalen Primärkulturen jene Signalwege zu identifizieren, die am Transport von Gephyrin, seiner Stabilisierung im postsynaptischen Netzwerk und seinem Abbau beteiligt sind. Im Verlauf der Arbeit konnte belegt werden, dass das Aktin-regulierende Phosphoprotein ena/VASP als Adapter die Interaktion von Gephyrin mit F-Aktin vermittelt, und dass diese Bindung ausreicht, um Gephyrin an das Aktin-Zytoskelett zu rekrutieren. Entgegen früheren Veröffentlichungen konnte die Bindung von ena/VASP im Bereich der sog. Linkerregion von Gephyrin nachgewiesen werden. Aktin-depolymerisierende Alkaloidbehandlungen von hippokampalen Neuronen bestätigten, das die Lokalisation von Gephyrin an sich entwickelnden inhibitorischen Kontakten von einem intakten Mikrofilamentsystem abhängig zu sein scheint. Das Aktin-Zytoskelett könnte somit eine transiente Rolle in der Ausbildung und Stabilisierung des Gephyrin-Netzwerkes in der frühen Entwicklung von inhibitorischen GABAergen Synapsen haben, während die Abhängigkeit der synaptisch-lokalisierten Gephyrincluster vom Aktin-Zytoskelett mit steigender neuronaler Differenzierung abnimmt. Zusätzlich konnte erstmals der Einfluss einzelner Signaltransduktionskaskaden auf die synaptische Lokalisation von Gephyrin nachgewiesen werden. Dabei hatte die Inhibition der Protein- Phosphatasen 1 und 2A eine Destabilisierung synaptisch-lokalisierter Gephyrincluster bei gleichzeitigem Anstieg der zytoplasmatischen Immunreaktivität zur Folge. Dies ist möglicherweise auf eine Hyperphosphorylierung wichtiger Sequenzabschnitten von Gephyrin zurückzuführen, wobei Änderungen im Phosphorylierungsstatus der Linkerregion von Gephyrin unter anderem die Assoziation mit dem Zytoskelett beeinträchtigen oder lösen könnten. Umgekehrt könnte eine Dephosphorylierung möglicherweise die Stabilität der Vernetzung erhöht. Kopräzipitationsstudien konnten zusätzlich nachweisen, dass Gephyrin, PP1 und PP2A nicht nur gemeinsam an inhibitorischen Synapsen vorliegen, sondern dass eine direkte Interaktion besteht. Dabei handelt es sich um den ersten Nachweis einer direkten Bindung von Gephyrin an Ser/Thr-Phosphatasen. Die Komplexbildung von PP1 und PP2A mit Gephyrin könnten der Regulation des Phosphorylierungsgrades dienen. Der genaue Mechanismus wird jedoch in weiteren Experimenten zu untersuchen sein.

Anisotrope Kolloide aus flüssigkristallinen Polymeren

In dieser Arbeit wird die Synthese, Charakterisierung und Manipulation anisotroper Kolloide aus flüssigkristallinen Polymeren beschrieben. Um Kolloide verschiedener Größe und aus verschiedenen Polymeren zu erhalten, wurden verschiedene Techniken verwendet. Einerseits wurden Kolloide aus nematischen und smektischen Polymeren mit Durchmessern meist im Bereich von 0,5 bis 3,5 Mikrometern hergestellt. Dazu wurden 16 verschiedene Acrylat- und Methacrylatmonomere synthetisiert und mittels Dispersionspolymerisation polymerisiert. Durch Variation der Polymerisationsbedingungen wurden Kolloide verschiedener Größe und Polydispersität erhalten. Durch Saatpolymerisation konnten zudem die Kugelgrößen bei gleichbleibend geringer Polydispersität erhöht werden. Polarisationsmikroskopie zeigt, dass die meisten Kolloide mit einer Größe zwischen ca. 2 bis 4 Mikrometern eine bipolare Direktorkonfiguration haben. Einige dieser Kolloide wurden mit einer optischen Pinzette mit zirkular polarisiertem Licht eingefangen und rotiert. Zum anderen wurden verschiedene flüssigkristalline Polymere (Polysiloxane, Hauptkettenpolymere und Polyacrylate) durch den Miniemulsionsprozess in Kolloide mit Durchmessern im Bereich von ca. 50 bis 300 nm überführt. Durch Variation der Emulgator- und Polymermenge sowie der Art des Emulgators konnte die Kugelgröße beeinflusst werden. Für die Polysiloxankolloide erfolgte die Aufklärung ihrer inneren Struktur mittels TEM und Kryo-TEM, da durch das Silizium im Polymerrückgrat ohne zusätzliches Anfärben ein Kontrast vorhanden ist. Die TEM-Aufnahmen zeigen deutlich die smektische Schichtstruktur innerhalb der Kolloide aus „verdünnten“ Copolysiloxanen und sind somit der erste direkte Beweis für die Mikrophasenseparation zwischen den Mesogenen und Polysiloxanketten, die bisher basierend auf Röntgenmessungen nur indirekt vorhergesagt wurde. Für die Copolysiloxane mit 2-Ring-Mesogenen wurden zwiebelartige Strukturen und für die Copolysiloxane mit 3-Ring-Mesogenen parallele Schichtstrukturen gefunden. Im ersten Fall folgt die smektische Schichtstruktur der Kugelsymmetrie des Kolloids, im zweiten Fall dominiert die Tendenz der smektischen Schichten, sich parallel anzuordnen.

Detektion neuroaktiver Substanzen durch Modifikation des Aktivitätsmusters neokortikaler Netzwerke auf Multielektrodenarrays – Charakterisierung und Optimierung eines neuronalen Biosensors

In dieser Arbeit wurden kortikale neuronale Netzwerke auf Multielektrodenarrays auf ihre Tauglichkeit als zellbasiertes Biosensorsystem untersucht. Der Schwerpunkt der pharmakologischen Untersuchungen an den ausgereiften kortikalen Netzwerken lag auf dem Einsatz von Substanzen, welche auf den GABAA-Rezeptor einwirken. Die Modifikation des spontan generierten Aktivitätsmusters ließ dabei Rückschlüsse auf die Wirksamkeit und den Wirkungsmechanismus der Testsubstanzen zu. Ferner war in den meisten Fällen eine Diskriminierung der auf den gleichen Rezeptor einwirkenden Substanzen möglich. Die Analyse der Spikerate und verschiedener auf Bursts beruhender Messparameter machte deutlich, dass die Burstrate bei den extrazellulären Ableitungen auf Netzwerkebene den sensitivsten und verlässlichsten Parameter zum Nachweis der Substanzeffekte darstellte. Durch die Verwendung kortikaler Netzwerke unter optimierten Kulturbedingungen und einer auf das System abgestimmten Analysesoftware konnte die Reproduzierbarkeit und Sensitivität im Vergleich zu anderen Studien deutlich verbessert werden. Um die extrazelluläre Signalableitung von einer möglichst geringen Zellanzahl und damit einem überschaubaren zellulären Netzwerk auf Multielektrodenarrays zu ermöglichen, wurden die Oberflächeneigenschaften der Substrate so modifiziert, dass die Lokalisation der Zellsomata und das Auswachsen der Neurite einer geometrischen Kontrolle unterlag. Die kontrollierte Substratbeschichtung des Adhäsionspromotors Poly-D-Lysin in einem triangulären Muster konnte dabei durch die Methode des Mikrokontaktstempelns realisiert werden. Durch das kontrollierte Zellwachstum konnte die extrazelluläre Ableitung von Netzwerken einer geringen Zelldichte über einen Zeitraum von mehreren Wochen ermöglicht werden. Die Untersuchung struktureller und morphogenetischer Eigenschaften, sowie elektrophysiologische Untersuchungen der strukturierten Netzwerke bewiesen, dass die kontrollierte Substratbeschichtung sich nicht negativ auf das Wachstum, die Synaptogenese und die Funktionalität auswirkte.