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Phage display selection of HIV specific conserved mimotopes with IgG from long-term non-progressors
(2005)
Poster presentation Background The aim of this study is to identify conserved epitopes of HIV-1 neutralizing antibodies in polyclonal plasma from LTNP to finally derive vaccine candidates. Materials and methods The presence of neutralizing antibodies in 9 LTNP sera was proved by in vitro neutralization assays. Phage displayed peptide libraries were screened with LTNP IgG. HIV-specific mimotopes were analyzed for homology to the gp120 structure by a software (3DEX) especially developed for this purpose. Mice were immunized with interesting phages and their sera were analyzed for neutralizing activities against HIV-1. Results After biopannings, between 19% and 75% HIV-specific phage clones were identified by ELISA. Mimotope sequences were identified and could be aligned by 3DEX to linear or conformational epitopes on gp120. A peptide specific immune response was detected in sera of immunized mice. The first mice sera analyzed showed neutralizing activities against HIV-1. Conclusion Mimotopes could be selected from LTNP sera that represent conformational epitopes on gp120. Those ones inducing neutralizing antibodies upon immunization potentially are suited to derive vaccine candidates.
Herman P. Schwan [1915–2005] was a distinguished scientist and engineer, and a founding father of the field of biomedical engineering. A man of integrity, Schwan influenced the lives of many, including his wife and children, and his many students and colleagues. Active in science until nearly the end of his life, he will be very much missed by his family and many colleagues.
We have isolated the human protein SNEV as downregulated in replicatively senescent cells. Sequence homology to the yeast splicing factor Prp19 suggested that SNEV might be the orthologue of Prp19 and therefore might also be involved in pre-mRNA splicing. We have used various approaches including gene complementation studies in yeast using a temperature sensitive mutant with a pleiotropic phenotype and SNEV immunodepletion from human HeLa nuclear extracts to determine its function. A human–yeast chimera was indeed capable of restoring the wild-type phenotype of the yeast mutant strain. In addition, immunodepletion of SNEV from human nuclear extracts resulted in a decrease of in vitro pre-mRNA splicing efficiency. Furthermore, as part of our analysis of protein–protein interactions within the CDC5L complex, we found that SNEV interacts with itself. The self-interaction domain was mapped to amino acids 56–74 in the protein's sequence and synthetic peptides derived from this region inhibit in vitro splicing by surprisingly interfering with spliceosome formation and stability. These results indicate that SNEV is the human orthologue of yeast PRP19, functions in splicing and that homo-oligomerization of SNEV in HeLa nuclear extract is essential for spliceosome assembly and that it might also be important for spliceosome stability.
The 5'-terminal cloverleaf (CL)-like RNA structures are essential for the initiation of positive- and negative-strand RNA synthesis of entero- and rhinoviruses. SLD is the cognate RNA ligand of the viral proteinase 3C (3Cpro), which is an indispensable component of the viral replication initiation complex. The structure of an 18mer RNA representing the apical stem and the cGUUAg D-loop of SLD from the first 5'-CL of BEV1 was determined in solution to a root-mean-square deviation (r.m.s.d.) (all heavy atoms) of 0.59 A (PDB 1Z30). The first (antiG) and last (synA) nucleotide of the D-loop forms a novel ‘pseudo base pair’ without direct hydrogen bonds. The backbone conformation and the base-stacking pattern of the cGUUAg-loop, however, are highly similar to that of the coxsackieviral uCACGg D-loop (PDB 1RFR) and of the stable cUUCGg tetraloop (PDB 1F7Y) but surprisingly dissimilar to the structure of a cGUAAg stable tetraloop (PDB 1MSY), even though the cGUUAg BEV D-loop and the cGUAAg tetraloop differ by 1 nt only. Together with the presented binding data, these findings provide independent experimental evidence for our model [O. Ohlenschläger, J. Wöhnert, E. Bucci, S. Seitz, S. Häfner, R. Ramachandran, R. Zell and M. Görlach (2004) Structure, 12, 237–248] that the proteinase 3Cpro recognizes structure rather than sequence.
Background: Costly structures need to represent an adaptive advantage in order to be maintained over evolutionary times. Contrary to many other conspicuous shell ornamentations of gastropods, the haired shells of several Stylommatophoran land snails still lack a convincing adaptive explanation. In the present study, we analysed the correlation between the presence/absence of hairs and habitat conditions in the genus Trochulus in a Bayesian framework of character evolution. Results: Haired shells appeared to be the ancestral character state, a feature most probably lost three times independently. These losses were correlated with a shift from humid to dry habitats, indicating an adaptive function of hairs in moist environments. It had been previously hypothesised that these costly protein structures of the outer shell layer facilitate the locomotion in moist habitats. Our experiments, on the contrary, showed an increased adherence of haired shells to wet surfaces. Conclusion: We propose the hypothesis that the possession of hairs facilitates the adherence of the snails to their herbaceous food plants during foraging when humidity levels are high. The absence of hairs in some Trochulus species could thus be explained as a loss of the potential adaptive function linked to habitat shifts.
Aims: This paper is a review of the literature on problem-related drinking of alcohol among medical doctors, and it deals with the epidemiology and results. Methods: A search of computer literature databases - PubMed and ETOH - was performed to locate articles reporting problem-related drinking among doctors, using population-based samples of doctors within the last two decades. Results: In the light of different definitions of problem-related drinking, there was found a breadth of prevalence of problem-related drinking - from heavy drinking and hazardous drinking (12%-16%) to misuse and dependence (6%-8%) - within the population-based samples of doctors. An increased risk was positively related to male doctors and doctors of the age of 40-45 years and older, and to some factors of work, lifestyle and health. Conclusion: For the future, it seems necessary to sensitise the research for problem-related drinking of doctors in Germany, e.g. initiating a representative survey, analysing the drinking of alcohol in the context of health, life-style and work-related factors.
We present a biologically-inspired system for real-time, feed-forward object recognition in cluttered scenes. Our system utilizes a vocabulary of very sparse features that are shared between and within different object models. To detect objects in a novel scene, these features are located in the image, and each detected feature votes for all objects that are consistent with its presence. Due to the sharing of features between object models our approach is more scalable to large object databases than traditional methods. To demonstrate the utility of this approach, we train our system to recognize any of 50 objects in everyday cluttered scenes with substantial occlusion. Without further optimization we also demonstrate near-perfect recognition on a standard 3-D recognition problem. Our system has an interpretation as a sparsely connected feed-forward neural network, making it a viable model for fast, feed-forward object recognition in the primate visual system.
In his paper, "The Canonization of German-language Digital Literature," Florian Hartling discusses "Net Literature," a relatively young phenomenon, that has its roots in experimental visual and concrete poetry and hypertext. With the use of new media technology, this new genre of literature has acquired much interest and is now considered to be one of the most important influences in contemporary art. Not only does Net Literature connect sound, video, and animation with interactivity and allows new forms of artistic expression, it also impacts significantly on the traditional functions of the literary system. Hartling suggests that, in relation to Net Literature, the notion of the "death of the author" gives birth to the "writing reader." Hartling presents the results of his study where he applies the concept of "canon" to German-language Net Literature and where he attempts to find out whether, in this new form of literature, a "canon" has already been formed. Based on Karl Erik Rosengren's framework of "mention technique," a sample of Germanlanguage reviews of Net Literature was analyzed. The study intends to test the applicability of Rosengren's method to the analysis of Net Literature, that is, whether it is valid to use a method that was originally developed for the empirical study of the traditional literary canon for the study of an emergent Net Literature.
Background: Depression is a disorder with high prevalence in primary health care and a significant burden of illness. The delivery of health care for depression, as well as other chronic illnesses, has been criticized for several reasons and new strategies to address the needs of these illnesses have been advocated. Case management is a patient-centered approach which has shown efficacy in the treatment of depression in highly organized Health Maintenance Organization (HMO) settings and which might also be effective in other, less structured settings. Methods/Design: PRoMPT (PRimary care Monitoring for depressive Patients Trial) is a cluster randomised controlled trial with General Practice (GP) as the unit of randomisation. The aim of the study is to evaluate a GP applied case-management for patients with major depressive disorder. 70 GPs were randomised either to intervention group or to control group with the control group delivering usual care. Each GP will include 10 patients suffering from major depressive disorder according to the DSM-IV criteria. The intervention group will receive treatment based on standardized guidelines and monthly telephone monitoring from a trained practice nurse. The nurse investigates the patient's status concerning the MDD criteria, his adherence to GPs prescriptions, possible side effects of medication, and treatment goal attainment. The control group receives usual care – including recommended guidelines. Main outcome measure is the cumulative score of the section depressive disorders (PHQ-9) from the German version of the Prime MD Patient Health Questionnaire (PHQ-D). Secondary outcome measures are the Beck-Depression-Inventory, self-reported adherence (adapted from Moriskey) and the SF-36. In addition, data are collected about patients' satisfaction (EUROPEP-tool), medication, health care utilization, comorbidity, suicide attempts and days out of work. The study comprises three assessment times: baseline (T0) , follow-up after 6 months (T1) and follow-up after 12 months (T2). Discussion: Depression is now recognized as a disorder with a high prevalence in primary care but with insufficient treatment response. Case management seems to be a promising intervention which has the potential to bridge the gap of the usually time-limited and fragmented provision of care. Case management has been proven to be effective in several studies but its application in the private general medical practice setting remains unclear.
This paper discusses constructions like “We’ll have two beers and a coffee.” that are typically used for beverage orders in restaurant contexts. We compare the behaviour of nouns in these constructions in three Germanic languages, English, Icelandic, and German, and take a closer look at the correlation of the morpho-syntactic and semantic-conceptual changes involved here. We show that even within such a closely related linguistic sample, one finds three different grammatical options for the expression of the same conceptual transition. Our findings suggest an analysis of coercion as a genuinely semantic phenomenon, a phenomenon that is located on a level of semantic representations that serves as an interface between the conceptual and the grammatical system and takes into account inter- and intralinguistic variations.