The potential of nanoscale carriers for drug delivery to intestinal mucosa and skin

Subjects of the present thesis were the formulation and evaluation of poly(lactide-co-glycolide) (PLGA) based nanoparticles (NP) addressing the biological barriers intestine and skin. As first step, fluorescence labelled PLGA NP were developed and characterized to study and to visualise interactions with such anatomical sites. Stable fluorescence labelling was accomplished by a covalent polymer modification with a fluoresceinamine. The potential of these NP to investigate penetration and storage in hair follicles in vitro and in vivo was examined within the scope of this work. Moreover, they were evaluated to represent powerful tools to study accumulation and retention in inflamed intestinal mucosa in inflammatory bowel diseases in future clinical studies. Then, the fluorescence labelled NP were advanced to dually fluorescence labelled NP by incorporation of Texas Red as fluorescent model drug. Those NP were applied to demonstrate the impact of multiphoton microscopy to simultaneously study penetration and drug release on excised human skin. Employing flufenamic acid as hydrophilic model drug, the influence of nanoencapsulation on drug penetration into skin was studied using PLGA NP as drug carriers. Finally, a technique to surface functionalize preformed PLGA NP was developed. This approach may allow subsequent versatile binding of proteins (targeting moieties, drugs) and dyes for various applications of interest e.g. targeting the intestinal mucosa.Das Thema der vorliegenden Dissertationsarbeit ist die Formulierung und Evaluierung von Nanopartikeln (NP) aus Polymilchsäure-co-glykolsäure (PLGA), die an der Darmschleimhaut und auf der Haut zum Einsatz kommen sollen. Zunächst wurden fluoreszenz-markierte PLGA NP hergestellt und charakterisiert, um damit Wechselwirkungen mit diesen biologischen Barrieren zu erforschen und zu visualisieren. Zur stabilen Fluoreszenzmarkierung wurde PLGA kovalent mit Fluoreszeinamin modifiziert. Im Rahmen dieser Arbeit wurde das Potential dieser NP aufgezeigt, Transport und Deposition in Haarfollikeln in vitro und in vivo zu untersuchen. Des Weiteren erwiesen dich diese NP als viel versprechend, um künftig eine Anreicherung und eine verlängerte Retention in entzündeter Darmschleimhaut bei Patienten mit chronisch entzündlichen Darmerkrankungen im Rahmen einer klinischen Studie zu überprüfen. Die beschriebene NP-Formulierung weiter entwickelt indem zusätzlich der Fluoreszenzfarbstoff Texas Red in die NP inkorporiert wurde, um als Modellarzneistoff zu dienen. Mit Hilfe dieser NP wurde gezeigt, dass es Multiphotonen-Mikroskopie ermöglicht, simultan Penetration und Arzneistofffreisetzung aus NP auf exzidierter Humanhaut zu visualisieren. Des Weiteren wurden PLGA NP dazu eingesetzt, um zu untersuchen, welchen Einfluss eine Verkapselung des lipophilen Modellarzneistoffs Flufenaminsäure auf den Arzneistofftransport in die Haut hat. Letztendlich wurde eine Technik entwickelt, mit Hilfe derer die Oberfläche von PLGA NP nach ihrer Formierung modifiziert werden kann. Dadurch können vielseitig Proteine wie "Targeting"-Komponenten oder Proteinarzneistoffe und Fluoreszenzfarbstoffe an NP gebunden werden

Przypadek tzw. wariantu choroby Niemanna-Picka typu Ci omówienie propozycji terapeutycznych

Niemann-Pick disease type C is a rare hereditary disorder caused by mutation-disrupted metabolism of cholesterol and low-density lipoprotein (LDL). In most patients, symptoms begin in childhood with severe clinical progression. We present a patient with heterozygote mutations 3001A>G and 3019C>G with late onset of the disease and positive response to treatment with miglustat. Behaviour and educational problems in childhood were probably related to the disease diagnosed later.Choroba Niemanna-Picka typu C jest rzadkim schorzeniem dziedziczonym autosomalnie recesywnie, spowodowanym zaburzeniem metabolizmu i transportu cholesterolu i LDL. Zachorowanie w większości przypadków dotyczy okresu dziecięcego i ma ciężki przebieg kliniczny. W artykule przedstawiono przypadek pacjenta z potwierdzoną złożoną mutacją 3001A>G i 3019C>G o późnym początku wystąpienia zaburzeń neurologicznych i korzystną odpowiedzią na leczenie miglustatem. Zaburzenia zachowania w wieku dziecięcym i trudności w nauce są najprawdopodobniej związane przyczynowo z później zdiagnozowaną chorobą

Unprecedented insights into extents of biological responses to physical forcing in an Arctic sub-mesoscale filament by combining high-resolution measurement approaches

In Fram Strait, we combined underway-sampling using the remote-controlled Automated Filtration System for Marine Microbes (AUTOFIM) with CTD-sampling for eDNA analyses, and with high-resolution optical measurements in an unprecedented approach to determine variability in plankton composition in response to physical forcing in a sub-mesoscale filament. We determined plankton composition and biomass near the surface with a horizontal resolution of ~ 2 km, and addressed vertical variability at five selected sites. Inside and near the filament, plankton composition was tightly linked to the hydrological dynamics related to the presence of sea ice. The comprehensive data set indicates that sea-ice melt related stratification near the surface inside the sub-mesoscale filament resulted in increased sequence abundances of sea ice-associated diatoms and zooplankton near the surface. In analogy to the physical data set, the underway eDNA data, complemented with highly sampled phytoplankton pigment data suggest a corridor of 7 km along the filament with enhanced photosynthetic biomass and sequence abundances of sea-ice associated plankton. Thus, based on our data we extrapolated an area of 350 km2 in Fram Strait with enhanced plankton abundances, possibly leading to enhanced POC export in an area that is around a magnitude larger than the visible streak of sea-ice

Surface Acoustic Wave (SAW) Resonators for Monitoring Conditioning Film Formation

We propose surface acoustic wave (SAW) resonators as a complementary tool for conditioning film monitoring. Conditioning films are formed by adsorption of inorganic and organic substances on a substrate the moment this substrate comes into contact with a liquid phase. In the case of implant insertion, for instance, initial protein adsorption is required to start wound healing, but it will also trigger immune reactions leading to inflammatory responses. The control of the initial protein adsorption would allow to promote the healing process and to suppress adverse immune reactions. Methods to investigate these adsorption processes are available, but it remains difficult to translate measurement results into actual protein binding events. Biosensor transducers allow user-friendly investigation of protein adsorption on different surfaces. The combination of several transduction principles leads to complementary results, allowing a more comprehensive characterization of the adsorbing layer. We introduce SAW resonators as a novel complementary tool for time-resolved conditioning film monitoring. SAW resonators were coated with polymers. The adsorption of the plasma proteins human serum albumin (HSA) and fibrinogen onto the polymer-coated surfaces were monitored. Frequency results were compared with quartz crystal microbalance (QCM) sensor measurements, which confirmed the suitability of the SAW resonators for this application

Finding needles in haystacks:Linking scientific names, reference specimens and molecular data for Fungi

DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Reannotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi.The Intramural Research Programs of the National Center for Biotechnology Information, National Library of Medicine and the National Human Genome Research Institute, both at the National Institutes of Health.http://www.ncbi.nlm.nih.gov/bioproject/PRJNA177353am201

REQUITE: A prospective multicentre cohort study of patients undergoing radiotherapy for breast, lung or prostate cancer

Purpose: REQUITE aimed to establish a resource for multi-national validation of models and biomarkers that predict risk of late toxicity following radiotherapy. The purpose of this article is to provide summary descriptive data. Methods: An international, prospective cohort study recruited cancer patients in 26 hospitals in eight countries between April 2014 and March 2017. Target recruitment was 5300 patients. Eligible patients had breast, prostate or lung cancer and planned potentially curable radiotherapy. Radiotherapy was prescribed according to local regimens, but centres used standardised data collection forms. Pre-treatment blood samples were collected. Patients were followed for a minimum of 12 (lung) or 24 (breast/prostate) months and summary descriptive statistics were generated. Results: The study recruited 2069 breast (99% of target), 1808 prostate (86%) and 561 lung (51%) cancer patients. The centralised, accessible database includes: physician-(47,025 forms) and patient-(54,901) reported outcomes; 11,563 breast photos; 17,107 DICOMs and 12,684 DVHs. Imputed genotype data are available for 4223 patients with European ancestry (1948 breast, 1728 prostate, 547 lung). Radiation-induced lymphocyte apoptosis (RILA) assay data are available for 1319 patients. DNA (n = 4409) and PAXgene tubes (n = 3039) are stored in the centralised biobank. Example prevalences of 2-year (1-year for lung) grade >= 2 CTCAE toxicities are 13% atrophy (breast), 3% rectal bleeding (prostate) and 27% dyspnoea (lung). Conclusion: The comprehensive centralised database and linked biobank is a valuable resource for the radiotherapy community for validating predictive models and biomarkers. Patient summary: Up to half of cancer patients undergo radiation therapy and irradiation of surrounding healthy tissue is unavoidable. Damage to healthy tissue can affect short-and long-term quality-of-life. Not all patients are equally sensitive to radiation "damage" but it is not possible at the moment to identify those who are. REQUITE was established with the aim of trying to understand more about how we could predict radiation sensitivity. The purpose of this paper is to provide an overview and summary of the data and material available. In the REQUITE study 4400 breast, prostate and lung cancer patients filled out questionnaires and donated blood. A large amount of data was collected in the same way. With all these data and samples a database and biobank were created that showed it is possible to collect this kind of information in a standardised way across countries. In the future, our database and linked biobank will be a resource for research and validation of clinical predictors and models of radiation sensitivity. REQUITE will also enable a better understanding of how many people suffer with radiotherapy toxicity

Qualitätsentwicklung an Ganztagsschulen

Durch die Verlagerung bzw. Stärkung von Entscheidungskompetenzen auf die bzw. der Ebene der Einzelschule wird es ermöglicht, Lösungs-/Gestaltungsansätze zu entwickeln, die auf die jeweiligen Bedürfnisse und Gegebenheiten vor Ort zugeschnitten werden können. Die kritische Auseinandersetzung mit den Erfahrungen anderer, die auf entsprechenden Fortbildungsveranstaltungen kommuniziert werden können, lässt Good-practice-Beispiele entstehen, aus denen sich Anregungen zur Realisierung eigener Vorhaben im Zuge der Ganztagsschulentwicklung ableiten lassen. Der dritte bayerische Ganztagsschulkongress "Qualitätsentwicklung an Ganztagsschulen" am 1. und 2. März 2012 in Forchheim bot den Teilnehmerinnen und Teilnehmern anhand diverser Vorträge, Workshops und Schulbesuchen die Möglichkeit zu Diskussion und Austausch. Der vorliegende Band dokumentiert die Veranstaltung

Finding needles in haystacks : linking scientific names, reference specimens and molecular data for Fungi

DNA phylogenetic comparisons have shown that morphology-based species recognition often underestimates fungal diversity. Therefore, the need for accurate DNA sequence data, tied to both correct taxonomic names and clearly annotated specimen data, has never been greater. Furthermore, the growing number of molecular ecology and microbiome projects using high-throughput sequencing require fast and effective methods for en masse species assignments. In this article, we focus on selecting and re-annotating a set of marker reference sequences that represent each currently accepted order of Fungi. The particular focus is on sequences from the internal transcribed spacer region in the nuclear ribosomal cistron, derived from type specimens and/or ex-type cultures. Reannotated and verified sequences were deposited in a curated public database at the National Center for Biotechnology Information (NCBI), namely the RefSeq Targeted Loci (RTL) database, and will be visible during routine sequence similarity searches with NR_prefixed accession numbers. A set of standards and protocols is proposed to improve the data quality of new sequences, and we suggest how type and other reference sequences can be used to improve identification of Fungi.The Intramural Research Programs of the National Center for Biotechnology Information, National Library of Medicine and the National Human Genome Research Institute, both at the National Institutes of Health.http://www.ncbi.nlm.nih.gov/bioproject/PRJNA177353am201

Differential predictors for alcohol use in adolescents as a function of familial risk

Abstract: Traditional models of future alcohol use in adolescents have used variable-centered approaches, predicting alcohol use from a set of variables across entire samples or populations. Following the proposition that predictive factors may vary in adolescents as a function of family history, we used a two-pronged approach by first defining clusters of familial risk, followed by prediction analyses within each cluster. Thus, for the first time in adolescents, we tested whether adolescents with a family history of drug abuse exhibit a set of predictors different from adolescents without a family history. We apply this approach to a genetic risk score and individual differences in personality, cognition, behavior (risk-taking and discounting) substance use behavior at age 14, life events, and functional brain imaging, to predict scores on the alcohol use disorders identification test (AUDIT) at age 14 and 16 in a sample of adolescents (N = 1659 at baseline, N = 1327 at follow-up) from the IMAGEN cohort, a longitudinal community-based cohort of adolescents. In the absence of familial risk (n = 616), individual differences in baseline drinking, personality measures (extraversion, negative thinking), discounting behaviors, life events, and ventral striatal activation during reward anticipation were significantly associated with future AUDIT scores, while the overall model explained 22% of the variance in future AUDIT. In the presence of familial risk (n = 711), drinking behavior at age 14, personality measures (extraversion, impulsivity), behavioral risk-taking, and life events were significantly associated with future AUDIT scores, explaining 20.1% of the overall variance. Results suggest that individual differences in personality, cognition, life events, brain function, and drinking behavior contribute differentially to the prediction of future alcohol misuse. This approach may inform more individualized preventive interventions