Species specific differences in use of ANP32 proteins by influenza A virus

Influenza A viruses (IAV) are subject to species barriers that prevent frequent zoonotic transmission and pandemics. One of these barriers is the poor activity of avian IAV polymerases in human cells. Differences between avian and mammalian ANP32 proteins underlie this host range barrier. Human ANP32A and ANP32B homologues both support function of human-adapted influenza polymerase but do not support efficient activity of avian IAV polymerase which requires avian ANP32A. We show here that the gene currently designated as avian ANP32B is evolutionarily distinct from mammalian ANP32B, and that chicken ANP32B does not support IAV polymerase activity even of human-adapted viruses. Consequently, IAV relies solely on chicken ANP32A to support its replication in chicken cells. Amino acids 129I and 130N, accounted for the inactivity of chicken ANP32B. Transfer of these residues to chicken ANP32A abolished support of IAV polymerase. Understanding ANP32 function will help develop antiviral strategies and aid the design of influenza virus resilient genome edited chickens

Ganzheitliche Untersuchungsmethoden zur Erfassung und Prüfung der Qualität ökologischer Lebensmittel: Stand der Entwicklung und Validierung

In dem wachsenden Markt ökologischer Lebensmittel werden Methoden zur produktorientierten Qualitätserfassung gefordert. Dabei geht es u.a. um die Unterscheidung von Produkten aus unterschiedlichen Anbauverfahren. Die Ziele des Projektes waren daher: 1. ausgewählte ganzheitliche Methoden gemäß ISO 17025 zu validieren, d.h. Laborprozesse festzulegen, sowie Einflussgrößen und Verfahrensmerkmale zu bestimmen, 2. zu testen, ob diese Verfahren eine Differenzierung von definierten Proben statistisch abgesichert zeigen können. . Diese Ziele konnten erreicht werden. Es wurde bestätigt, dass einige der Methoden auf Grundlage dokumentierter Prozeduren Lebensmittel aus definierten Anbauversuchen (u.a. aus dem DOK-Versuch am FIBL/CH) reproduzierbar unterscheiden können. Die Koordination und die Validierung der Kupferchlorid-Kristallisation sowie die Messung der Polyphenole lag bei der Universität Kassel, FG Ökologische Lebensmittelqualität und Ernährungskultur. Die KWALIS GmbH, Dipperz, validierte die Fluoreszenz-Anregungsspektroskopie und die Bestimmung des Physiologischen Aminosäurestatus, die EQC GmbH, Weidenbach die elektrochemischen Messungen. Dr. Kromidas, Saarbrücken übernahm die Beratung der Validierungsprozeduren. . An Blindproben wurde untersucht, ob die Verfahren für Weizen- und Möhrenproben aus definierten Anbau- und Sortenversuchen geeignet sind (Fragestellung der Validierung). Die Proben wurden von unabhängiger Stelle (OEL-FAL, Trenthorst) codiert. Die Proben wurden gleichzeitig an alle Partner versandt; dadurch konnten die Methoden auch untereinander verglichen werden. Die Methoden Kupferchlorid-Kristallisation, Fluoreszenz-Anregungsspektroskopie und Physiologischer Aminosäurestatus sind für die Fragestellung geeignet. Mit allen drei Methoden konnten die Proben differenziert und gruppiert werden. Darüber hinaus konnten mit der Fluoreszenz-Anregungsspektroskopie und über den physiologischen Aminosäurestatus die Proben auch den Anbauweisen richtig zugeordnet werden. Allerdings ist damit noch keine Aussage über die Fähigkeit dieser Verfahren möglich, generell Proben aus ökologischer und konventioneller Herkunft zu unterscheiden. Dafür sind weitere Untersuchungen sowohl an Proben definierter Herkunft als auch an Marktproben notwendig

ANP32 proteins are essential for influenza virus replication in human cells

ANP32 proteins have been implicated in supporting influenza virus replication, but most of the work to date has focused on the ability of avian Anp32 proteins to overcome restriction of avian influenza polymerases in human cells. Using a CRISPR approach we show that human ANP32A and ANP32B are functionally redundant but essential host factors for mammalian-adapted influenza A virus (IAV) and influenza B virus (IBV) replication in human cells. When both proteins are absent from human cells, influenza polymerases are unable to replicate the viral genome, and infectious virus cannot propagate. Provision of exogenous ANP32A or –B recovers polymerase activity and virus growth. We demonstrate that this redundancy is absent in the murine Anp32 orthologues: murine Anp32A is incapable of recovering IAV polymerase activity, while murine Anp32B can. Intriguingly, IBV polymerase is able to use murine Anp32A. We show using a domain swap and point mutations that the LRR 5 region comprises an important functional domain for mammalian ANP32 proteins. Our approach has identified a pair of essential host factors for influenza virus replication and can be harnessed to inform future interventions

The Structure and Delivery of Police Use of Force Training: A German Case Study

The current study aims to investigate the current structure and delivery of police recruit training. Using a case study approach, we systematically observed a semester of police training that consisted of 30 h with a specific focus on police use of force training. Field notes and time-on-task data was analysed using an inductive approach. The results revealed, first, a lack of constructive alignment of the training modules and learning tasks within the training settings. Second, an adherence to traditional linear approaches to training resulting in high amounts of augmented instruction and feedback and a one-size-fits all approach to technical and tactical behaviour. Third, a non-efficient use of available training time with low amounts of engagement in representatively designed tasks that stimulated problem-solving processes. Based on these results we suggest that there is a need: (a) for police trainers and curriculum designers to align the objectives, practice structure and delivery of police training with the needs of police officers in the field (e.g. conflict resolution); (b) for police trainers to employ more learner-centred pedagogical approaches that account for individual action capabilities and resources, and allow for high amounts of training time with representatively designed training tasks; and (c) for senior managers of overall police training decision-makers to provide the necessary trainer education, in order to furnish trainers with the knowledge and tools to appropriately plan, deliver and reflect upon their practice in keeping with concept of constructive alignment

Foodways in transition: food plants, diet and local perceptions of change in a Costa Rican Ngäbe community

Background Indigenous populations are undergoing rapid ethnobiological, nutritional and socioeconomic transitions while being increasingly integrated into modernizing societies. To better understand the dynamics of these transitions, this article aims to characterize the cultural domain of food plants and analyze its relation with current day diets, and the local perceptions of changes given amongst the Ngäbe people of Southern Conte-Burica, Costa Rica, as production of food plants by its residents is hypothesized to be drastically in recession with an decreased local production in the area and new conservation and development paradigms being implemented. Methods Extensive freelisting, interviews and workshops were used to collect the data from 72 participants on their knowledge of food plants, their current dietary practices and their perceptions of change in local foodways, while cultural domain analysis, descriptive statistical analyses and development of fundamental explanatory themes were employed to analyze the data. Results Results show a food plants domain composed of 140 species, of which 85 % grow in the area, with a medium level of cultural consensus, and some age-based variation. Although many plants still grow in the area, in many key species a decrease on local production–even abandonment–was found, with much reduced cultivation areas. Yet, the domain appears to be largely theoretical, with little evidence of use; and the diet today is predominantly dependent on foods bought from the store (more than 50 % of basic ingredients), many of which were not salient or not even recognized as ‘food plants’ in freelists exercises. While changes in the importance of food plants were largely deemed a result of changes in cultural preferences for store bought processed food stuffs and changing values associated with farming and being food self-sufficient, Ngäbe were also aware of how changing household livelihood activities, and the subsequent loss of knowledge and use of food plants, were in fact being driven by changes in social and political policies, despite increases in forest cover and biodiversity. Conclusions Ngäbe foodways are changing in different and somewhat disconnected ways: knowledge of food plants is varied, reflecting most relevant changes in dietary practices such as lower cultivation areas and greater dependence on food from stores by all families. We attribute dietary shifts to socioeconomic and political changes in recent decades, in particular to a reduction of local production of food, new economic structures and agents related to the State and globalization

CXCR4 expression on circulating pan-cytokeratin positive cells is associated with survival in patients with advanced non-small cell lung cancer

<p>Abstract</p> <p>Background</p> <p>The CXC chemokine, CXCL12, and its receptor, CXCR4 promote metastases of a variety of solid tumors, including non-small cell lung cancer (NSCLC). The expression of CXCR4 on tumor cells may represent a critical biomarker for their propensity to metastasize. This study was performed to evaluate the hypothesis that co-expression of pan-cytokeratin and CXCR4 may be a prognostic marker for patients with advanced NSCLC.</p> <p>Methods</p> <p>We evaluated CXCR4 levels on circulating pan-cytokeratin positive cells from patients with NSCLC. NSCLC tumor and metastases were also assessed for the presence of CXCR4.</p> <p>Results</p> <p>Pan-cytokeratin positive cells were increased in the circulation of patients with NSCLC, as compared to normal control subjects. Patients with pan-cytokeratin +/CXCR4+ = 2,500 cells/ml had a significant improvement in median survival when compared with patients with pan-cytokeratin +/CXCR4+ >2,500 cells/ml (not achieved versus 14 weeks). CXCR4 expression was found on NSCLC tumors and at sites of tumor metastasis.</p> <p>Conclusion</p> <p>This study suggests that CXCR4 may be a prognostic marker in NSCLC, and provides hypothesis-generating results, which may be important in determining metastatic potential. In future studies, we will prospectively evaluate the prognostic significance of pan-cytokeratin/CXCR4+ cells, and determine the mechanisms involved in the regulation of CXCR4 expression on tumor cells in a larger patient population.</p

The Max b-HLH-LZ Can Transduce into Cells and Inhibit c-Myc Transcriptional Activities

The inhibition of the functions of c-Myc (endogenous and oncogenic) was recently shown to provide a spectacular therapeutic index in cancer mouse models, with complete tumor regression and minimal side-effects in normal tissues. This was achieved by the systemic and conditional expression of omomyc, the cDNA of a designed mutant of the b-HLH-LZ of c-Myc named Omomyc. The overall mode of action of Omomyc consists in the sequestration of Max and the concomitant competition of the Omomyc/Max complex with the endogenous c-Myc/Max heterodimer. This leads to the inhibition of the transactivation of Myc target genes involved in proliferation and metabolism. While this body of work has provided extraordinary insights to guide the future development of new cancer therapies that target c-Myc, Omomyc itself is not a therapeutic agent. In this context, we sought to exploit the use of a b-HLH-LZ to inhibit c-Myc in a cancer cell line in a more direct fashion. We demonstrate that the b-HLH-LZ domain of Max (Max*) behaves as a bona fide protein transduction domain (PTD) that can efficiently transduce across cellular membrane via through endocytosis and translocate to the nucleus. In addition, we show that the treatment of HeLa cells with Max* leads to a reduction of metabolism and proliferation rate. Accordingly, we observe a decrease of the population of HeLa cells in S phase, an accumulation in G1/G0 and the induction of apoptosis. In agreement with these phenotypic changes, we show by q-RT-PCR that the treatment of HeLa cells with Max* leads to the activation of the transcription c-Myc repressed genes as well as the repression of the expression of c-Myc activated genes. In addition to the novel discovery that the Max b-HLH-LZ is a PTD, our findings open up new avenues and strategies for the direct inhibition of c-Myc with b-HLH-LZ analogs