Isoform-spezifische Funktionen mitogen-aktivierter Proteinkinasen in Transkriptionskontrolle und Proliferation

In vielen humanen Neoplasien findet sich eine erhöhte Aktivität des Raf-Mek-Erk-Signaltransduktionsweges. Zunächst wurde davon ausgegangen, dass diese erhöhte Aktivität hauptsächlich durch die Ras-Onkogene hervorgerufen wird. Doch mittlerweile konnten auch Mutationen der Raf Gene in humanen Neoplasien nachgewiesen werden. Gegen Raf und Mek konnten eine Anzahl von Enzyminhibitoren entwickelt werden. Der Nachteil vieler dieser Inhibitoren ist, dass sie nicht zwischen den einzelnen Kinaseisoformen unterscheiden können. In dieser Arbeit ist es nun das erste Mal gelungen, jede Komponente des Raf-Mek-Erk-Signaltransduktionsweges einzeln mittels RNA Interferenz effizient zu inhibieren. Dabei konnte die Rolle der verschiedenen Isoformen in der Proliferation, Morphologie und Genex-pression von transformierten Zellen definiert werden. In den NIH3T3-pEJ Zellen konnte A-Raf erstmals eine antiapoptotische Rolle zugewiesen werden. Diese Hemmung der Apoptose läuft möglicherweise über einen Mek2-abhängigen Weg und ist an die Mitochondrien gekoppelt. Für die beiden Mek Kinasen konnten unter-schiedliche Funktionen in der Signalweiterleitung gezeigt werden. Mek2 spielt die Hauptrolle in der Aktivierung der beiden Substratkinasen Erk1 und Erk2. Der Verlust der Mek1 Isoform wird dagegen möglicherweise durch eine erhöhte Expression von Mek2 kompensiert und wirkt sich nicht so stark auf die Phosphorylierung von Erk1/2 aus. Durch die Verwendung von Erk1 und Erk2 spezifischen siRNAs konnte eine Trennung zwischen der Proliferationsre-gulation und der Kontrolle der morphologischen Transformation herausgearbeitet werden. Durch die Verwendung von Mikroarrays ist es gelungen, beiden Phänotypen ein Genexpres-sionsprofil zuzuordnen. Neben Unterschieden zwischen den verschiedenen Kinaseisoformen konnten neue, potentielle Feedbacks beschrieben werden.In many human neoplasia an increased activity of the RAF/MEK/ERK- signaling pathway is found. First it was assumed that this raised activity is caused primarily by the RAS onco-genes. However, meanwhile mutations in the RAF genes could be also proved in human neo-plasia. A number of enzyme inhibitors have been developed against the RAF and MEK pro-teins. The disadvantage of many of these inhibitors is that they cannot distinguish between the different kinase isoforms. In this work it has succeeded the first time to inhibit every compo-nent of the RAF/MEK/ERK- signaling pathway individually by means of interference RNA. Beside this, the role of the different isoforms in the proliferation, morphology and genetic profile of transformed cells could be defined. For the first time A-Raf could be assigned an anti-apoptotic role in NIH3T3-pEJ cells. This inhibition of the apoptosis possibly runs through a Mek2-dependent way and is coupled to the mitochondria. For both Mek kinases different functions could be shown in the downstream signaling. Mek2 plays the leading role in the activation of both downstream kinases Erk1 and Erk2. The loss of the Mek1 isoform expression is possibly compensated through an increased expression of Mek2 and does not affect the phosphorylation of Erk1 / 2 so strongly. A discri-mination between the regulation of proliferation and the control of the morphological trans-formation could be worked out by the use of Erk1 and Erk2 specific siRNAs. By the use of micorarray an expression profile of both phenotypes has assigned. Beside differences between the different kinases new, potential feedback pathways could be described

Skythisches Gold in griechischem Stil : Untersuchungen zur nordpontischen Toreutik am Beispiel der Waffen- und Gefäßbeigaben des Solocha-Kurgans

Die vorliegende Arbeit beschäftigt sich mit Edelmetallarbeiten, die aus skythischen Bestattungen des nördlichen Schwarzmeergebiets stammen. Ausgangspunkt für die Untersuchungen bilden die Funde aus dem Solocha-Kurgan, der in den Jahren 1912/13 von dem russischen Archäologen N.I. Veselovskij ausgegraben wurde. Nach einem einleitenden Kapitel zur allgemeinen Forschungsgeschichte der sogenannten graeco-skythischen Toreutik wird im zweiten Kapitel der Solocha-Kurgan vorgestellt. Von den beiden Katakombengräbern, die Veselovskij öffnete, datiert die zentrale Bestattung in die Zeit um 400 v. Chr., das Seitengrab um 370 v. Chr. Aufgrund der Beraubung der zentralen Bestattung steht vor allem die reiche Beigabenausstattung der ungestörten seitlichen Katakombe im Vordergrund der Betrachtung. In Einzeluntersuchungen werden die Edelmetallarbeiten – der berühmte Kamm, Gefäße und Prunkwaffen - zunächst im Kontext ihrer jeweiligen Gattung betrachtet. Hierfür werden die Stücke jeder Gattung vorgestellt und das jeweilige Objekt aus dem Solocha-Kurgan vor diesem Hintergrund diskutiert. Zur Veranschaulichung dienen tabellarische Zusammenstellungen von Gefäßausstattungen und Prunkwaffen in skythischen Bestattungen des 5. – 4. Jhs. v. Chr. Die Gesamtbetrachtung zeigt, dass sich einige der Edelmetallfunde aus dem Solocha-Kurgan gut in die allgemeine Entwicklung der Edelmetallbeigaben in skythischen Bestattungen einfügen. Dazu gehören die importierten Silbergefäße, die mit Appliken versehenen Holzschalen sowie das Trinkhorn aus der seitlichen Katakombe. Auch der dort gefundene Prunkakinakes besitzt Vorgänger im skythischen Material. Ein Novum hinsichtlich Form und Dekor stellen dagegen der berühmte Goldkamm, die kugeligen Silberflaschen, die Silberschalen mit den segmentförmigen Griffen sowie der großformatige Silberbeschlag des Goryts aus der Seitenbestattung dar. Obwohl sie teilweise an frühere Formen anknüpfen, stellen sie eigenständige Gestaltungen dar. Aufgrund figürlicher Darstellungen im Stil des sogenannten ethnographischen Realismus können einige Stücke enger zusammengeschlossen werden und sind wohl als Erzeugnisse einer einzelnen Auftragsarbeit zu deuten. Aufgrund der fehlenden Vorläufer ist es eher unwahrscheinlich, dass diese Arbeiten von einem Skythen in Auftrag gegeben werden konnten. Die anfangs noch geringe Anzahl derartiger Objekte spricht darüber hinaus auch gegen eine Interpretation als Handelsware. Da analog zur Verbreitung der späteren Erzeugnisse graeco-skythischer Toreutik von einer Fertigung im Bosporanischen Reich - am ehesten in Pantikapaion - ausgegangen werden kann, scheint folgende Erklärung am plausibelsten: Die Prunkobjekte aus dem Solocha-Kurgan stellen „politische“ Geschenke dar, die der bosporanische Herrscher Leukon I. (389/88-349/48 v. Chr.) in Auftrag gab und an einen skythischen Verbündeten übermittelte. Dementsprechend wären die in den Darstellungen gegeneinander kämpfenden Skythen als Verbündete und Gegner des Bosporanischen Reiches zu erklären. Als Grundlage für die Bearbeitung der einzelnen toreutischen Werke dient ein ausführlich kommentierter Katalog der Fundkomplexe, aus denen die Objekte stammen. Hierbei gibt der Bestattungszeitraum einen ersten Anhaltspunkt für die Datierung der Edelmetallarbeiten, weiter können aus ihrer Lage innerhalb der Bestattung und der Vergesellschaftung mit anderen Fundstücken Rückschlüsse auf ihre Funktion gezogen werden. Daneben wurde für die einzelnen Gegenstände ein Objektkatalog getrennt nach Gattungen angelegt, in dem der Aufbewahrungsort sowie Maße, Datierung, Beschreibung und Literatur angegeben sind.Scythian Gold in greek style : Studies on northpontic toreutics based on the vessels and armament from the Solokha-Kurgan The thesis deals with gold and silver objects which were found in Scythian kurgans of the northern Black Sea region. In the focus stand the finds from the Solokha Kurgan which was excavated in 1912/13 by the Russian archaeologist N.I. Veselovskij. An introductory chapter deals with the history of research on the so called Graeco-Scythian toreutics, the Solokha Kurgan is presented in the second chapter. Under the burial mound Veselovskij opened two catacomb graves; the central burial is dated to the end of the 5th century BC, the second grave in the south west part of the kurgan to about 370 BC. Because the central tomb was almost entirely robbed, the attention is directed particularly on the rich grave goods from the undisturbed lateral catacomb. In individual studies the different objects - the different vessels, the well known comb and the decorative weapons – are first seen in the context of their respective genres (chap. 2-4). For this purpose, the pieces for each genre are described in chronological order and the particular object from the Solokha Kurgan is discussed against this background. Tabular compilations illustrate the equipment of Scythian burials of the 5th and 4th century BC with vessels and weapons made of precious metals. The overall analysis shows that some of the finds from the Solokha Kurgan fit well into the general development of precious grave goods in Scythian burials. This refers to the imported silver vessels, the wooden bowls with gold appliques and the drinking horn from the side catacomb. Also the akinakes which was found there has predecessors in Scythian finds. However, a novelty in form and decoration represent the gold comb, the spherical silver bottles, the silver bowls with the segment-shaped handles and the large-sized silver overlay of the gorytos from the side funeral. Although these things are linked in part to earlier scythian forms, they represent new created designs. Some of the pieces can be grouped together because of their figural representations in the style of the so-called ethnographic realism; probably they have to be interpreted as the products of a single commission. Due to the lack of precursors, it is unlikely that these pieces were ordered by a Scythian noble. The initially small number of such objects also speaks against an interpretation as normal trade goods. By analogy with the spread of the somewhat later objects of Graeco-Scythian toreutics a production in the Bosporan kingdom - most likely in Pantikapaion - can be expected. So the following explanation seems to be most plausible: The precious metal objects from the Solokha-Kurgan probably were "political" gifts which Leukon I., the ruler of the Bosporan kingdom from 389/88 to 349/48 BC commissioned and gave to a Scythian ally. Accordingly, the fighting Scythians of the battle scenes of the comb and the gorytos overlay are to be interpreted as allies and opponents of the Bosporan Kingdom. As basis for the study serves a detailed annotated catalogue of scythian burial mounds from which the objects originated. The funeral date gives a first hint at the dating of the objects of precious metals; their position within the burials and the combination with other finds provide clues to their function. In a second catalogue the gold and silver objects are described with measurements, dating, and literature. (Translation: A. Wieland/G. Wahl)Or scythe dans un style grec : Recherches sur la toreutique des régions du Pont-Euxin septentrional: les offrandes funéraires du kourgane de Solokha Cette étude aborde les oeuvres en métal précieux provenant des sépultures scythes du littoral nord de la Mer Noire. Les trouvailles du kourgane de Solokha, qui fut fouillé en 1912-13 par l'archéologue russe N.I. Veselovskij, en constituent le point de départ. L'introduction dresse l'état de la recherche dans le domaine de la toreutique « gréco-scythe ». Elle est suivie d'un chapitre présentant le kourgane de Solokha dont deux tombes furent mises au jour par Veselovskij : la sépulture principale, située au milieu du kourgane et datée des environs de 400 av. J.- C., et la sépulture latérale, datée vers 370 av. J.-C. C'est le riche mobilier métallique de la seconde sépulture - trouvée intacte, contrairement à la première qui était violée- qui est ici présenté en détail. Les objets en métal précieux du kourgane de Solokha – le fameux peigne, la vaisselle et les armes d'apparat – sont étudiés au sein de leurs catégories typologiques. Confrontée aux pièces du même genre qui sont également discutées, chaque oeuvre est ainsi placée dans son contexte typologique. La présentation est complétée par des tableaux synthétiques regroupant les vases et les armes d'apparat accompagnant des sépultures scythes des Ve et IVe siècles av. J.-C. Cette vue d'ensemble montre que certains parmi les objets en métal précieux du kourgane de Solokha s'insèrent bien dans l'évolution générale des offrandes funéraires luxueuses trouvées dans des sépultures scythes : la vaisselle d'argent importée, les vases à boire en bois décorés d'appliques ainsi que le rhyton de la tombe latérale en font partie tout comme l'akinakès précieux de la même sépulture, qui a des antécédents au sein du matériel scythe. En revanche, le célèbre peigne d'or et les objets en argent, tels les vases à panse sphérique, les coupes à anses arrondies et le grand revêtement de goryte de la tombe latérale sont nouveaux autant pour leur forme que pour leur décor. Tout en restant en partie rattachées à des formes antérieures, ces oeuvres témoignent d'un caractère original. En raison de leurs compositions figurées dans un style connu sous le nom de « réalisme ethnographique », certaines oeuvres peuvent être regroupées dans un ensemble restreint et doivent être issues d'une seule commande. Cependant, l'absence d'antécédents rend leur association avec un commanditaire scythe peu probable. Par ailleurs, le nombre à l'origine limité de ce type d'objets empêche de les interpréter comme des produits de commerce. Au contraire, la large diffusion des plus récentes créations de la toreutique « gréco-scythe » laisse présumer une fabrication locale, dans le royaume du Bosphore – plus probablement à Pantikapaion. Ainsi, l'hypothèse suivante semble la plus plausible pour expliquer ce phénomène : les oeuvres luxueuses du kourgane de Solokha représenteraient des cadeaux diplomatiques que le roi du Bosphore Leukon Ier (389/88-349/48 av. J.-C.) aurait commandités afin de les offrir à un de ses alliés scythes. A cet égard, les Scythes en combat figurant dans les scènes représentées devraient être interprétés comme des alliés et des ennemis du royaume du Bosphore. Le recensement des sépultures et des ensembles funéraires sous forme de catalogue commenté est fourni à l'appui des arguments avancés au sujet du mobilier toreutique. Ainsi, les datations proposées pour une sépulture livrent un premier indice pour la datation des objets en métal précieux qui l'accompagnaient, tandis que leur emplacement à l'intérieur de la tombe tout comme leur relation aux autres offrandes permettent de mieux cerner leur fonction. Un second catalogue regroupe les objets classés par genre avec les informations sur le lieu de leur conservation, leurs dimensions, la datation, la description et la bibliographie. (Traduction : K. Charatzopoulou

Interaction Effects of Child Weight Status and Parental Feeding Practices on Children’s Eating Disorder Symptomatology

(1) Background: Research on parental feeding practices and non-normative eating behavior including loss of control (LOC) eating and eating disorder psychopathology indicated separate associations of these variables with child weight status, especially in early childhood. This study cross-sectionally examined interaction effects of restriction, monitoring, pressure to eat, and children’s weight status on disordered eating in children aged 8–13 years. (2) Methods: A population-based sample of N = 904 children and their mothers completed the Eating Disorder Examination Questionnaire for Children and the Child Feeding Questionnaire. Child anthropometrics were objectively measured. Hierarchical linear and logistic regression analyses were conducted for cross-sectionally predicting global eating disorder psychopathology and recurrent LOC eating by feeding practices and child weight status for younger (8–10 years) and older (11–13 years) ages. (3) Results: Restriction x Child weight status significantly predicted global eating disorder psychopathology in younger children and recurrent LOC eating in older children. Monitoring x Child weight status significantly predicted eating disorder psychopathology in older children. A higher versus lower child weight status was associated with adverse eating behaviors, particularly in children with mothers reporting high restriction and monitoring. (4) Conclusions: Detrimental associations between higher child weight status and child eating disorder symptomatology held especially true for children whose mothers strongly control child food intake

Interview-based assessment of avoidant/restrictive food intake disorder (ARFID): A pilot study evaluating an ARFID module for the Eating Disorder Examination

Objective Although avoidant/restrictive food intake disorder (ARFID) has been included as a new diagnostic entity of childhood feeding and eating disorders, there is a lack of measures to reliably and validly assess ARFID. In addition, virtually nothing is known about clinical characteristics of ARFID in nonclinical samples. Method The present study presents the development and validation of an ARFID module for the child and parent version of the Eating Disorder Examination (EDE) in a nonclinical sample of N = 39 children between 8 and 13 years with underweight and/or restrictive eating behaviors. For evaluating the ARFID module's reliability, the convergence of diagnoses between two independent raters and between the child and parent module was determined. The module's validity was evaluated based on the full-length child version of the EDE, a 24 h food record, parent-reported psychosocial functioning and self-reported quality of life, and objective anthropometric measures. Results In total, n = 7 children received an ARFID diagnosis. The ARFID module showed high interrater reliability, especially for the parent version, and high convergence between child and parent report. Evidence for the module's convergent, divergent, and discriminant validity was provided. Specifically, children with versus without ARFID reported significantly less macro- and micronutrient intake and were more likely to be underweight. Discussion This pilot study indicates the child and parent version of the EDE ARFID module to be promising for diagnosing ARFID in a structured way but still necessitates a validation in a larger clinical and community-based sample

Characteristics of avoidant/restrictive food intake disorder in a general pediatric inpatient sample

Objective: Although patients with avoidant/restrictive food intake disorder (ARFID) often consult general pediatric services initially, existing literature mostly concentrated on ntensive eating disorder treatment settings. This cross-sectional study sought to describe symptoms of ARFID and their associations with eating disorder psychopathology, quality of life, anthropometry, and physical comorbidities in a general pediatric sample. Methods: In N = 111 patients (8-18 years) seeking treatment for physical diseases, prevalence of ARFID-related restrictive eating behaviors was estimated by self-report and compared to population-based data (N = 799). Using self-report and medical record data, further ARFID diagnostic criteria were evaluated. Patients with versus without symptoms of ARFID based on self-report and medical records were compared in diverse clinical variables. Results: The prevalence of self-reported symptoms of ARFID was not higher in the inpatient than population-based sample. Only picky eating and shape concern were more common in the inpatient than population-based sample. Although 69% of the inpatient sample reported any restrictive eating behaviors, only 7.2% of patients showed symptoms of ARFID based on medical records in addition to self-report, particularly those with underweight, without significant effects for age, sex, and medical diagnoses. Discussion: The study revealed the importance of considering ARFID within the treatment of children and adolescents with physical diseases, especially for those with underweight. Further research is needed to replicate the findings with interview-based measures and to investigate the direction of effects in ARFID and its physical correlates

Media Use of Mothers, Media Use of Children, and Parent–Child Interaction Are Related to Behavioral Difficulties and Strengths of Children

The present study investigated the associations of media use of children, media use of mothers, and parent-child interactions with behavioral strengths and difficulties in children. Screen time of 553 2- to 9-year-old children and their mothers were indicated by the daily durations of their TV/games console/computer/mobile phone use. The amount of parent–child interaction was indicated by the frequencies of shared activities at home. Behavioral strengths and difficulties of children were investigated using the Strengths and Difficulties Questionnaire. Children whose mothers reported high screen times (>/= 5 h/day) were significantly more likely to show high screen times (>/= 2 h/day). High screen time of children was associated with more conduct problems, more symptoms of hyperactivity/inattention and less prosocial behavior. High screen time of mothers was associated with emotional problems, conduct problems, and symptoms of hyperactivity/inattention. In contrast, a higher frequency of parent–child interactions was associated with fewer conduct problems, fewer peer-relationship problems, and more prosocial behavior of children. Children might use the media behavior of their mothers as a role model for their own media use. Furthermore, the findings suggest that media use of children and mothers and parent–child interaction contribute independently to behavioral strengths and difficulties of children

An 8-item short form of the Eating Disorder Examination-Questionnaire adapted for children (ChEDE-Q8)

Objective: Eating disturbances are common in children placing a vulnerable group of them at risk for full-syndrome eating disorders and adverse health outcomes. In order to provide a valid self-report assessment of eating disorder psychopathology in children, a short form of the child version of the Eating Disorder Examination (ChEDE-Q) was psychometrically evaluated. Similar to the EDE-Q, the ChEDE-Q provides assessment of eating disorder psychopathology related to anorexia nervosa, bulimia nervosa, and binge-eating disorder. However, it does not assess symptoms of avoidant/restrictive food intake disorder, pica, or rumination disorder. Method: In 1836 participants ages 7 to 18 years, recruited from two independent population-based samples, the factor structure of the recently established 8-item short form EDE-Q8 for adults was examined, including measurement invariance analyses on age, gender, and weight status derived from objectively measured weight and height. For convergent validity, the ChEDE-Q global score, Body Esteem Scale, Strengths and Difficulties Questionnaire, and sociodemographic characteristics were used. Item characteristics and age- and gender-specific norms were calculated. Results: Confirmatory factor analysis revealed good model fit for the 8-item ChEDE-Q. Measurement invariance analyses indicated strict invariance for all analyzed subgroups. Convergent validity was provided through associations with well-established questionnaires and age, gender, and weight status, in expected directions. Discussion: The newly developed ChEDE-Q8 proved to be a psychometrically sound and economical self-report assessment tool of eating disorder psychopathology in children. Further validation studies are needed, particularly concerning discriminant and predictive validity

Satiety regulation in children with loss of control eating and attention-deficit/hyperactivity disorder: a test meal study

Children with loss of control (LOC) eating and attention-deficit/hyperactivity disorder (ADHD) are at risk for excessive weight gain. However, it is unclear whether or not these children show disturbances in hunger and satiety regulation. The goal was to examine the food intake and sense of LOC over eating as well as LOC eating-related characteristics during test meal in children with LOC eating and ADHD. Children aged 8-13 y with LOC eating (n = 33), ADHD (n = 32), and matched healthy controls (n = 33), consumed a test meal consisting of their chosen lunch food, with the instruction to eat until feeling full. Sense of LOC over eating, desire to eat, feelings of hunger, and liking of food were repeatedly assessed during test meal. Children with LOC eating and ADHD did not show a higher food intake at maximum satiety compared to control children. Sense of LOC over eating was significantly higher in children with LOC eating compared to children with ADHD and matched controls. Secondary analyses revealed that children with LOC eating ate marginally faster than control children. Both children with LOC eating and ADHD reported greater desire to eat, feelings of hunger, and liking of food during test meal than control children. Even though the results did not reveal statistical evidence to support the assumption of a disturbed food intake in children with LOC eating and ADHD, LOC eating related characteristics were significantly higher in these children compared to the control children. Sense of LOC over eating was confirmed as a specific characteristic of LOC eating. The examination of behavioral indicators of hunger and satiety dysregulation should be complemented with physiological indicators in future research

Macro- and Micronutrient Intake in Children with Avoidant/Restrictive Food Intake Disorder

Although case studies in avoidant/restrictive food intake disorder (ARFID) indicate severe nutritional deficiencies in those with a highly limited amount or variety of food intake, systematic analyses on food intake in treatment-seeking children and adolescents with ARFID are lacking. Within this study, n = 20 patients with an interview-based diagnosis of ARFID (0–17 years) were included and compared to n = 20 healthy controls individually matched for age and sex. Children or parents completed three-day food diaries and a food list. Macronutrient, vitamin, and mineral supply was determined based on the percentage of their recommended intake. The results showed a significantly lower total energy and protein intake in ARFID versus controls, with trends for lower fat and carbohydrate intake. ARFID subtypes of limited amount versus variety of food intake significantly differed in macro-, but not micronutrient intake. Those with ARFID met only 20–30% of the recommended intake for most vitamins and minerals, with significantly lower intake relative to controls for vitamin B1, B2, C, K, zinc, iron, and potassium. Variety of food intake was significantly reduced in ARFID versus controls in all food groups except carbohydrates. This study demonstrated that ARFID goes along with reduced everyday life macro- and micronutrient intake, which may increase the risk for developmental and health problems. Future studies additionally assessing serum nutrient levels in a larger sample may further explore differences in food intake across diverse ARFID presentations

Trimeric G-proteins of the trans-Golgi network are involved in the formation of constitutive secretory vesicles and immature secretory granules

AbstractNon-hydrolysable analogues of GTP, such as GTPγS and GMP-PNP, have previously been shown to inhibit the formation of constitutive secretory vesicles (CSVs) and immature secretory granules (ISGs) from the trans-Golgi network (TGN). Using a cell-free system, we show here that the formation of these vesicles is also inhibited by [AIF4], a compound known to act on trimeric G-proteins. Addition of highly purified G-protein βγ subunits stimulated, in a differential manner, the cell-free formation of both CSVs and ISGs. ADP-ribosylation experiments revealed the presence of a pertussis toxin-sensitive G-protein α subunit in the TGN. We conclude that trimeric G-proteins regulate the formation of secretory vesicles from the TGN