Possible role of SIRT1 and SIRT3 in post-translational modifications in human breast milk during the neonatal period

Amino Acids - We measured free and proteinic concentrations of native and modified amino acids from post-translational modifications (PTMs) and correlated them with the activity of SIRT1 and SIRT3...     

Two novel effectors of trafficking and maturation of the yeast plasma membrane H+‐ATPase

The endoplasmic reticulum (ER) is the entry site of proteins into the endomembrane system. Proteins exit the ER via coat protein II (COPII) vesicles in a selective manner, mediated either by direct interaction with the COPII coat or aided by cargo receptors. Despite the fundamental role of such receptors in protein sorting, only a few have been identified. To further define the machinery that packages secretory cargo and targets proteins from the ER to Golgi membranes, we used multiple systematic approaches, which revealed 2 uncharacterized proteins that mediate the trafficking and maturation of Pma1, the essential yeast plasma membrane proton ATPase. Ydl121c (Exp1) is an ER protein that binds Pma1, is packaged into COPII vesicles, and whose deletion causes ER retention of Pma1. Ykl077w (Psg1) physically interacts with Exp1 and can be found in the Golgi and coat protein I (COPI) vesicles but does not directly bind Pma1. Loss of Psg1 causes enhanced degradation of Pma1 in the vacuole. Our findings suggest that Exp1 is a Pma1 cargo receptor and that Psg1 aids Pma1 maturation in the Golgi or affects its retrieval. More generally our work shows the utility of high content screens in the identification of novel trafficking components.      

Morphologische Untersuchungen zur Proliferationskinetik der Mäusehaut

Zusammenfassung 1. Es wird eine Methode zur Herstellung, Montierung und Färbung von epidermalen Häutchenpräparaten zur dreidimensionalen Analyse des Epidermis-Aufbaues beschrieben. Eine Kombination mit der Histoautoradiographie ist bedingt möglich. Damit ist eine Basis für weitere Untersuchungen mit neuer Fragestellung geschaffen. 2. Die untersuchten Objekte Mäuseohr und Rückenhaut zeigen eine bisher unbekannte, auffallende Musterbildung mit Zuordnung bestimmter interfollikulärer basaler Gruppen zu Zellen des Stratum spinosum und des Stratum corneum. Es handelt sich um rosettenähnliche Gebilde aus mehreren peripheren Basalzellen und meist einer zentralen Spinosumzelle. 3. Die Bedeutung der Befunde für die normale wie für die pathologische Regeneration und Neoplasie der Epidermis ist offen. Es werden Beziehungen zum Regenerationsmodus und damit zur Proliferationskinetik der Epidermis diskutiert. In diesem Zusammenhang sind die Untersuchungen auch für die Humanpathologie relevant.

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Morphologic investigations of the proliferation kinetics of the mouse epidermisStandardized methods and results

Summary 1. A method for the preparation, mounting, and staining of thin preparations of mouse epidermis is described, which allows a three-dimensional analysis of epidermal structure. A limited combination of this technique with histo-autoradiography is possible, thereby presenting a broader basis for further investigations. 2. Both ear and back skin show a striking, previously unrecognized pattern of organization, in which definite interfollicular basal cell groups are oriented towards cells of the stratum spinosum and stratum corneum. A rosettelike picture of many peripheral basal cells and, in most instances, a central spinosum cell is found. 3. The significance of our findings for normal and pathological regenerations as well as neoplasia of the epidermis remains open. We discuss relationships to the mode of regeneration and thus the kinetics of proliferation in the epidermis. Our investigations are also relevant for human pathology in this regard.

Meinem Vater Kurt Goerttler zur Vollendung des 75. Lebensjahres herzlichst zugeeignet.     

Developing the Autism Model of Implementation for Autism spectrum disorder community providers: study protocol

ABSTRACT: BACKGROUND: Currently, 1 out of 88 children are diagnosed with an autism spectrum disorder (ASD), and the estimated cost for treatment services is $126 billion annually. Typically, ASD community providers (ASD-CPs) provide services to children with any severity of ASD symptoms using a combination of various treatment paradigms, some with an evidence-base and some without. When evidence-based practices (EBPs) are successfully implemented by ASD-CPs, they can result in positive outcomes. Despite this promise, EBPs are often implemented unsuccessfully and other treatments used by ASD-CPs lack supportive evidence, especially for school-age children with ASD. While it is not well understood why ASD-CPs are not implementing EBPs, organizational and individual characteristics likely play a role. As a response to this need and to improve the lives of children with ASD and their families, this study aims to develop and test the feasibility and acceptability of the Autism Model of Implementation (AMI) to support the implementation of EBPs by ASD-CPs. Methods/design An academic-community collaboration developed to partner with ASD-CPs will facilitate the development of the AMI, a process specifically for use by ASD community-based agencies. Using a mixed methods approach, the project will assess agency and individual factors likely to facilitate or hinder implementing EBPs in this context; develop the AMI to address identified barriers and facilitators; and pilot test the AMI to examine its feasibility and acceptability using a specific EBP to treat anxiety disorders in school-age children with ASD. DISCUSSION: The AMI will represent a data-informed approach to facilitate implementation of EBPs by ASD-CPs by providing an implementation model specifically developed for this context. This study is designed to address the real-world implications of EBP implementation in ASD community-based agencies. In doing so, the AMI will help to provide children with ASD the best and most effective services in their own community. Moreover, the proposed study will positively impact the field of implementation science by providing an empirically supported and tested model of implementation to facilitate the identification, adoption, and use of EBPs.     

Opsonizing properties of an isolated hemolymph agglutinin and demonstration of lectin-like recognition molecules at the surface of hemocytes fromMytilus edulis

Summary Mytilus hemolymph was found to contain an agglutinin which could be inhibited by mucin. The agglutinin was isolated by affinity chromatography using neuraminidase-treated mucin/Sepharose.In vitro phagocytosis experiments revealed that only about 5% of washed hemocytes phagocytosed yeast cells suspended in a Tris-buffered NaCl-solution, whereas yeast suspended in hemolymph was normally ingested by more than 50% of the hemocytes. This relatively high phagocytic activity was shown to depend on the presence of two serum factors: When purified agglutinin was added to saline-suspended yeast, phagocytosis rates returned to normal, demonstrating opsonizing properties of the purified agglutinin. — On the other hand, addition of Ca⁺⁺-ions to saline caused an increase of the phagocytic activity of hemocytes. This was interpreted to indicate the activation of divalent cation-dependent recognition molecules at the hemocyte surface. The function of these postulated recognition factors was demonstrated by phagocytosis inhibition tests. Their location at the hemocyte membrane became evident by binding of specific antiagglutinin IgG purified by help of an agglutinin/Sepharose column from an antiserum raised againstMytilus serum proteins. Consequently, humoral as well as cell bound agglutinin molecules are involved in the attachment of yeast cells toMytilus hemocytes which subsequently internalize foreign cells.Abbreviations DAB dimethylamino benzaldehyde - PO peroxidase - IgG immunoglobulin G     

Cloning and characterization of an adenoviral vector for highly efficient and doxycycline – suppressible expression of bioactive human single – chain interleukin 12 in colon cancer

Background  

Interleukin-12 (IL-12) is well characterized to induce cellular antitumoral immunity by activation of NK-cells and T-lymphocytes. However, systemic administration of recombinant human IL-12 resulted in severe toxicity without perceptible therapeutic benefit. Even though intratumoral expression of IL-12 leads to tumor regression and long-term survival in a variety of animal models, clinical trials have not yet shown a significant therapeutic benefit. One major obstacle in the treatment with IL-12 is to overcome the relatively low expression of the therapeutic gene without compromising the safety of such an approach. Our objective was to generate an adenoviral vector system enabling the regulated expression of very high levels of bioactive, human IL-12.