Testing the length limit of loop grafting in a helical repeat protein.

Alpha-helical repeat proteins such as consensus-designed tetratricopeptide repeats (CTPRs) are exceptionally stable molecules that are able to tolerate destabilizing sequence alterations and are therefore becoming increasingly valued as a modular platform for biotechnology and biotherapeutic applications. A simple approach to functionalize the CTPR scaffold that we are pioneering is the insertion of short linear motifs (SLiMs) into the loops between adjacent repeats. Here, we test the limits of the scaffold by inserting 17 highly diverse amino acid sequences of up to 58 amino acids in length into a two-repeat protein and examine the impact on protein folding, stability and solubility. The sequences include three SLiMs that bind oncoproteins and eleven naturally occurring linker sequences all predicted to be intrinsically disordered but with conformational preferences ranging from compact globules to expanded coils. We show that the loop-grafted proteins retain the native CTPR structure and are thermally stable with melting temperatures above 60 ​°C, despite the longest loop sequence being almost the same size as the CTPR scaffold itself (68 amino acids). Although the main determinant of the effect of stability was found to be loop length and was relatively insensitive to amino acid composition, the relationship between protein solubility and the loop sequences was more complex, with the presence of negatively charged amino acids enhancing the solubility. Our findings will help us to fully realize the potential of the repeat-protein scaffold, allowing a rational design approach to create artificial modular proteins with customized functional capabilities

Familien mit einem schizophren erkrankten Elternteil: Ergebnisse einer fallrekonstruktiven Familienstudie

"Fragestellung: In diesem Beitrag werden Ergebnisse einer fallrekonstruktiven Studie vorgestellt, bei der Familien mit einem schizophren erkrankten Elternteil befragt wurden. Dabei sollte untersucht wurden, wie sich die Schizophrenie auf die Familienmitglieder, ihren Alltag und ihre Beziehungen auswirkt. Methodik: Die Auswertung erfolgte sowohl fall- als auch themenbezogen, wobei inhaltsanalytische und fallrekonstruktive Verfahren eingesetzt wurden. Ergebnisse: Trotz der Vielfalt der familiären Konstellationen und Belastungslagen zeigte sich eine Reihe charakteristischer Muster. Kinder zu haben bedeutet für erkrankte Eltern, sowohl Ressourcen als auch Belastungen zu haben. Paar- und Familienbeziehungen sind oft stark beeinträchtigt und weisen ein hohes Risiko für Beziehungsabbrüche auf. Zugleich wird im Familienalltag eine Normalität jenseits der Erkrankung angestrebt und erlebt. Normalisierungs- und Vermeidungsstrategien können dazu beitragen, dass die Erkrankung zu einem Tabuthema wird. Viele Kinder sind daher unzureichend über die elterliche Schizophrenie informiert. Sie sind in dieser Situation oft überfordert und entwickeln ihrerseits Verhaltensauffälligkeiten, Ängste und Depressionen. Diskussion: Die Schizophrenie kann insofern als 'Familienerkrankung' gedeutet werden, als sie das gesamte Familiensystem beeinflusst, belastet und gefährdet. Aus diesem Grund sollten verstärkt familienorientierte Präventionsangebote bereitgestellt werden, wobei Gesundheitswesen und Jugendhilfe eng miteinander kooperieren sollten." (Autorenreferat)"Objective: This study aims at investigating the impact of a parental schizophrenia on the family members, their everyday life and their relations. For this purpose, we conduct qualitative interviews with mothers and fathers suffering from schizophrenia, their spouses and children. Methods: Interview data is analyzed using case-reconstructive as well as content analysis methods. Results: Although results illustrate a great variety of family constellations and burdening circumstances, there are a number of typical patterns: Having children is perceived by affected parents in an ambiguous manner, i.e. as a resource as well as a distress. Relationships of couples and families are often impaired, resulting in a high risk of abandonment of relationships. At the same time, family members strive for normality in everyday life. Normalisation and avoidance strategies can bring about that the schizophrenia becomes a taboo issue within the family. Thus, with regard to their parent's illness, many of the children are insufficiently informed. Often, the children are overstrained by this situation and, in turn, may develop behaviour disorders, anxiety, or depression. Discussion: In sum, schizophrenia can be considered as a 'family disease' as it strongly affects the whole family system. Hence, it is necessary to provide preventive help offers for affected parents, their spouses and children. For delivering support, youth welfare and public health services should cooperate closely." (author's abstract

Dynamics of novel photoactive AgCl microstars and their environmental applications

In the field of micromotors many efforts have been taken to find a substitute for peroxide as fuel. While most approaches turn towards other toxic high energy chemicals such as hydrazine, we introduce here an energy source that is widely used in nature: light. Light is an ideal source of energy and some materials, such as AgCl, have the inherent property to transform light energy for chemical processes, which can be used to achieve propulsion. In the case of silver chloride, one process observed after light exposure is surface modification, which leads to the release of ions, generating chemo-osmotic gradients. Here we present endeavors to use those processes to propel uniquely shaped micro-objects of microstar morphology with a high surfaceto- volume ratio, study their dynamics and present approaches to go towards real environmental applications

A large fraction of HLA class I ligands are proteasome-generated spliced peptides

The proteasome generates the epitopes presented on human leukocyte antigen (HLA) class I molecules that elicit CD8(+) T cell responses. Reports of proteasome-generated spliced epitopes exist, but they have been regarded as rare events. Here, however, we show that the proteasome-generated spliced peptide pool accounts for one-third of the entire HLA class I immunopeptidome in terms of diversity and one-fourth in terms of abundance. This pool also represents a unique set of antigens, possessing particular and distinguishing features. We validated this observation using a range of complementary experimental and bioinformatics approaches, as well as multiple cell types. The widespread appearance and abundance of proteasome-catalyzed peptide splicing events has implications for immunobiology and autoimmunity theories and may provide a previously untapped source of epitopes for use in vaccines and cancer immunotherapy

Niedertiefenbach. Ein Galeriegrab der spätneolithischen Wartberggruppe südwestlich von Niedertiefenbach (Landkreis Limburg-Weilburg, Hessen)

Das Galeriegrab von Niedertiefenbach stellt einen besonderen Befund der späten Wartberggruppe im Neolithikum der Deutschen Mittelgebirgsschwelle dar; nur hier findet sich ein in zehn Straten geborgenes Bestattungskollektiv von mindestens 177 Individuen. Eine umfangreiche Serie von Radiokarbondatierungen belegt ein wesentlich höheres Alter des Grabes als bisher publiziert und erlaubt die Modellierung einer Bestattungssequenz zwischen 3.350 und 2.900 cal BC. Eine neue paläopathologische Untersuchung, vorerst nur an den Schädeln vorgenommen, zeigt einen hohen Krankenstand mit zahlreichen entzündlichen Prozessen. Mit einer verheilten Trepanation bei einem Kind wird aber zugleich auch ein besonderes Wissen im Bereich der medizinischen Versorgung deutlich. Erste Analysen zur aDNA mittels Hochfrequenzsequenzierung (HTS) belegen die Anwesenheit mitochondrialer DNA mit den Haplogruppen U5 und X2. Der erstmalige Einsatz eines CAD/CAM-Computersystems (CEREC) bietet eine detaillierte Erfassung morphologischer und pathologischer Merkmale an den Zähnen und Kiefern mit noch folgender metrisch-statistischer Analyse

The programme on ecosystem change and society (PECS)–a decade of deepening social-ecological research through a place-based focus

The Programme on Ecosystem Change and Society (PECS) was established in 2011, and is now one of the major international social-ecological systems (SES) research networks. During this time, SES research has undergone a phase of rapid growth and has grown into an influential branch of sustainability science. In this Perspective, we argue that SES research has also deepened over the past decade, and helped to shed light on key dimensions of SES dynamics (e.g. system feedbacks, aspects of system design, goals and paradigms) that can lead to tangible action for solving the major sustainability challenges of our time. We suggest four ways in which the growth of place-based SES research, fostered by networks such as PECS, has contributed to these developments, namely by: 1) shedding light on transformational change, 2) revealing the social dynamics shaping SES, 3) bringing together diverse types of knowledge, and 4) encouraging reflexive researchers

Promoter methylation of CDKN2A and lack of p16 expression characterize patients with hepatocellular carcinoma

<p>Abstract</p> <p>Background</p> <p>The product of CDKN2A, p16 is an essential regulator of the cell cycle controlling the entry into the S-phase. Herein, we evaluated CDKN2A promoter methylation and p16 protein expression for the differentiation of hepatocellular carcinoma (HCC) from other liver tumors.</p> <p>Methods</p> <p>Tumor and corresponding non-tumor liver tissue samples were obtained from 85 patients with liver tumors. CDKN2A promoter methylation was studied using MethyLight technique and methylation-specific PCR (MSP). In the MethyLight analysis, samples with ≥ 4% of PMR (percentage of methylated reference) were regarded as hypermethylated. p16 expression was evaluated by immunohistochemistry in tissue sections (n = 148) obtained from 81 patients using an immunoreactivity score (IRS) ranging from 0 (no expression) to 6 (strong expression).</p> <p>Results</p> <p>Hypermethylation of the CDKN2A promoter was found in 23 HCCs (69.7%; mean PMR = 42.34 ± 27.8%), six (20.7%; mean PMR = 31.85 ± 18%) liver metastases and in the extralesional tissue of only one patient. Using MSP, 32% of the non-tumor (n = 85), 70% of the HCCs, 40% of the CCCs and 24% of the liver metastases were hypermethylated. Correspondingly, nuclear p16 expression was found immunohistochemically in five (10.9%, mean IRS = 0.5) HCCs, 23 (92%; mean IRS = 4.9) metastases and only occasionally in hepatocytes of non-lesional liver tissues (mean IRS = 1.2). The difference of CDKN2A-methylation and p16 protein expression between HCCs and liver metastases was statistically significant (p < 0.01, respectively).</p> <p>Conclusion</p> <p>Promoter methylation of CDKN2A gene and lack of p16 expression characterize patients with HCC.</p