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Berlin Brandenburg

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  • 1
    Language: English
    In: Proceedings of the National Academy of Sciences of the United States of America, 24 August 2010, Vol.107(34), pp.15022-6
    Description: Gene targeting by homologous recombination in embryonic stem cells is extensively used to generate specific mouse mutants. However, most mammalian species lack tools for targeted gene manipulation. Since double-strand breaks strongly increase the rate of homologous recombination at genomic loci, we explored whether gene targeting can be directly performed in zygotes by the use of zinc-finger nucleases. Here we report that gene targeting is achieved in 1.7-4.5% of murine one-cell embryos upon the coinjection of targeting vectors with zinc-finger nucleases, without preselection. These findings enable the manipulation of the mammalian germ line in a single step in zygotes, independent of ES cells.
    Keywords: Endonucleases -- Metabolism ; Gene Targeting -- Methods ; Zygote -- Metabolism
    ISSN: 00278424
    E-ISSN: 1091-6490
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  • 2
    Language: English
    In: Proceedings of the National Academy of Sciences of the United States of America, 05 March 2013, Vol.110(10), pp.3782-7
    Description: The study of genetic disease mechanisms relies mostly on targeted mouse mutants that are derived from engineered embryonic stem (ES) cells. Nevertheless, the establishment of mutant ES cells is laborious and time-consuming, restricting the study of the increasing number of human disease mutations discovered by high-throughput genomic analysis. Here, we present an advanced approach for the production of mouse disease models by microinjection of transcription activator-like effector nucleases (TALENs) and synthetic oligodeoxynucleotides into one-cell embryos. Within 2 d of embryo injection, we created and corrected chocolate missense mutations in the small GTPase RAB38; a regulator of intracellular vesicle trafficking and phenotypic model of Hermansky-Pudlak syndrome. Because ES cell cultures and targeting vectors are not required, this technology enables instant germline modifications, making heterozygous mutants available within 18 wk. The key features of direct mutagenesis by TALENs and oligodeoxynucleotides, minimal effort and high speed, catalyze the generation of future in vivo models for the study of human disease mechanisms and interventions.
    Keywords: Disease Models, Animal ; Germ-Line Mutation ; Genetic Diseases, Inborn -- Genetics ; Oligodeoxyribonucleotides -- Administration & Dosage
    ISSN: 00278424
    E-ISSN: 1091-6490
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  • 3
    Language: English
    In: Journal of Community Health, 2011, Vol.36(4), pp.669-674
    Description: To describe the participatory approach used to develop “Good For The Neighborhood” (GFTN), a community program to improve the health of four underserved communities. A core program was developed involving a “park and stay” approach to impact four underserved predominately minority communities (two predominately African American, 1 predominately Latino, and the Seneca Nation of Indians). The core program includes health screenings, risk assessments, health education, and exposure to health services. An extensive tracking and evaluation system was developed to determine participation and impact on the community. Multi-methods (key informant interviews, focus groups, surveys) were implemented to gain feedback from community partners and participants as to how to adopt the program to meet the needs of the community. GFTN has been sustained for over 3 years and has reached over 3,500 predominately minority individuals in four communities with 1/3 of participants engaging regularly in the program. The program has evolved in the four communities to meet specific needs. A “park and stay” approach in partnership with the community has led to a strong program that community partners and residents embrace. Community ownership and social networking, including word-of-mouth from residents is essential to establishing a successful program.
    Keywords: Community health ; Health disparities ; Evaluation ; Community based participatory research ; Minorities
    ISSN: 0094-5145
    E-ISSN: 1573-3610
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  • 4
    Language: English
    In: American journal of health behavior, July 2016, Vol.40(4), pp.523-33
    Description: We investigated the relationship between financial literacy and patient engagement while considering the possible interaction effects due to patient financial responsibility and patient-physician shared decision making, and the impact of personal attributes. Participants consisted of an Internet-based sample of American adults (N = 160). Hierarchical multiple linear regression analysis was conducted to examine the relationship of the study variables on patient engagement. We found that patient financial responsibility (β = -.19, p 〈 .05) and patient-physician shared decision-making (β = .17, p 〈 .05) predicted patient engagement. However, there was no statistically significant relationship between patient financial literacy and patient engagement; moreover, the moderation effects of patient financial responsibility and shared decision making with financial literacy also were not statistically significant. Increasing patient financial responsibility and patient-physician shared decision making can impact patient engagement. Understanding the predictors of patient engagement and the factors that influence financial behaviors may allow for the development of interventions to enable patients to make better healthcare decisions, and ultimately, improve health outcomes.
    Keywords: Financing, Personal ; Patient Participation -- Economics
    ISSN: 10873244
    E-ISSN: 1945-7359
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  • 5
    Language: English
    In: PloS one, 2015, Vol.10(2), pp.e0118405
    Description: Multiresistant nosocomial pathogens often cause life-threatening infections that are sometimes untreatable with currently available antibiotics. Staphylococci and enterococci are the predominant Gram-positive species associated with hospital-acquired infections. These infections often lead to extended hospital stay and excess mortality. In this study, a panel of fully human monoclonal antibodies was isolated from a healthy individual by selection of B-cells producing antibodies with high opsonic killing against E. faecalis 12030. Variable domains (VH and VL) of these immunoglobulin genes were amplified by PCR and cloned into an eukaryotic expression vector containing the constant domains of a human IgG1 molecule and the human lambda constant domain. These constructs were transfected into CHO cells and culture supernatants were collected and tested by opsonophagocytic assay against E. faecalis and S. aureus strains (including MRSA). At concentrations of 600 pg/ml, opsonic killing was between 40% and 70% against all strains tested. Monoclonal antibodies were also evaluated in a mouse sepsis model (using S. aureus LAC and E. faecium), a mouse peritonitis model (using S. aureus Newman and LAC) and a rat endocarditis model (using E. faecalis 12030) and were shown to provide protection in all models at a concentration of 4 μg/kg per animal. Here we present a method to produce fully human IgG1 monoclonal antibodies that are opsonic in vitro and protective in vivo against several multiresistant Gram-positive bacteria. The monoclonal antibodies presented in this study are significantly more effective compared to another monoclonal antibody currently in clinical trials.
    Keywords: Antibodies, Monoclonal -- Isolation & Purification ; Drug Resistance, Multiple, Bacterial -- Immunology ; Enterococcus Faecalis -- Drug Effects ; Staphylococcus Aureus -- Drug Effects
    E-ISSN: 1932-6203
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  • 6
    Language: English
    In: PLoS ONE, 01 January 2016, Vol.11(2), p.e0149053
    Description: This study aimed to assess the association of clinical factors with P2Y12-dependent platelet inhibition as monitored by the ratio of ADP- to TRAP-induced platelet aggregation and conventional ADP-induced aggregation, respectively.Controversial findings to identify and overcome high platelet reactivity (HPR) after coronary stent-implantation and to improve clinical outcome by tailored anti-platelet therapy exist. Monitoring anti-platelet therapy ex vivo underlies several confounding parameters causing that ex vivo platelet aggregation might not reflect in vivo platelet inhibition.In a single centre observational study, multiple electrode aggregometry was performed in whole blood of patients after recent coronary stent-implantation. Relative ADP-induced aggregation (r-ADP-agg) was defined as the ratio of ADP- to TRAP- induced aggregation reflecting the individual degree of P2Y12-mediated platelet reactivity.Platelet aggregation was assessed in 359 patients. Means (± SD) of TRAP-, ADP-induced aggregation and r-ADP-agg were 794 ± 239 AU*min, 297 ± 153 AU*min and 37 ± 14%, respectively. While ADP- and TRAP-induced platelet aggregation correlated significantly with platelet count (ADP: r = 0.302; p〈0.001; TRAP: r = 0.509 p〈0.001), r-ADP-agg values did not (r = -0.003; p = 0.960). These findings were unaltered in multivariate analyses adjusting for a range of factors potentially influencing platelet aggregation. The presence of an acute coronary syndrome and body weight were found to correlate with both ADP-induced platelet aggregation and r-ADP-agg.The ratio of ADP- to TRAP-induced platelet aggregation quantifies P2Y12-dependent platelet inhibition independently of the platelet count in contrast to conventional ADP-induced aggregation. Furthermore, r-ADP-agg was associated with the presence of an acute coronary syndrome and body weight as well as ADP-induced aggregation. Thus, the r-ADP-agg is a more valid reflecting platelet aggregation and potentially prognosis after coronary stent-implantation in P2Y12-mediated HPR than conventional ADP-induced platelet aggregation.
    Keywords: Sciences (General)
    E-ISSN: 1932-6203
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  • 7
    In: Scientific Reports, 2015, Vol.5
    Description: The role of microglia in amyloid-β (Aβ) deposition is controversial. In the present study, an organotypic hippocampal slice culture (OHSC) system with an in vivo-like microglial-neuronal environment was used to investigate the potential contribution of microglia to Aβ plaque formation. We found that microglia ingested Aβ, thereby preventing plaque formation in OHSCs. Conversely, Aβ deposits formed rapidly in microglia-free wild-type slices. The capacity to prevent Aβ plaque formation was absent in forebrain microglia from young adult but not juvenile 5xFamilial Alzheimer's disease (FAD) mice. Since no loss of Aβ clearance capacity was observed in both wild-type and cerebellar microglia from 5xFAD animals, the high Aβ1-42 burden in the forebrain of 5xFAD animals likely underlies the exhaustion of microglial Aβ clearance capacity. These data may therefore explain why Aβ plaque formation has never been described in wild-type mice, and point to a beneficial role of microglia in AD pathology. We also describe a new method to study Aβ plaque formation in a cell culture setting.
    Keywords: Alzheimer Disease -- Pathology ; Amyloid Beta-Peptides -- Metabolism ; Hippocampus -- Pathology ; Microglia -- Pathology ; Peptide Fragments -- Metabolism ; Plaque, Amyloid -- Pathology ; Prosencephalon -- Pathology;
    ISSN: 20452322
    E-ISSN: 20452322
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  • 8
    Language: English
    In: Thrombosis Research, February 2016, Vol.138, pp.63-68
    Description: Novel (or non-vitamin K antagonist) oral anti-coagulants (NOACs) are antagonists of coagulation factors (F) Xa (rivaroxaban) or IIa (dabigatran), and their non-inferiority compared with vitamin K antagonists has been demonstrated in patients with non-valvular atrial fibrillation. However, it is still not fully understood if and how dabigatran and rivaroxaban impact platelet function. This observational study aimed to assess platelet function in patients receiving dabigatran or rivaroxaban. This was a single centre, observational study quantifying platelet aggregation in 90 patients treated with NOACs by multiple electrode aggregometry. The thrombin receptor activating peptide (TRAP)-induced platelet aggregation was significantly higher in 35 patients receiving dabigatran (d) compared with control (c) patients (d 108 ± 31 vs. c 85 ± 30 arbitrary units [AU]∗ min, p 〈 0.001). Patients receiving rivaroxaban (r) showed no differences compared with the control group (r 88 ± 32 vs. c 85 ± 30 AU ∗ min, p = 0.335). In intraindividual time courses of 16 patients, a significantly higher aggregation was found after the administration of dabigatran (before vs. after; 83 ± 29 vs. 100 ± 31 AU ∗ min, p = 0.009). In this observational study, the TRAP-induced platelet aggregation was enhanced in cardiovascular patients receiving dabigatran. This might be explained by a change in the expression profile of thrombin receptors on the surface of platelets. Rivaroxaban had no influence on platelet aggregation.
    Keywords: Dabigatran ; Rivaroxaban ; Platelet ; Aggregation ; Medicine
    ISSN: 0049-3848
    E-ISSN: 1879-2472
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  • 9
    Language: English
    In: Forensic Science International, 10 October 2012, Vol.222(1-3), pp.256-258
    Description: The HSP70 superfamily is a reliable biomarker for hyperthermia, hypothermia and hypoxia. The Enzyme-linked Immunosorbent Assay (ELISA) respectively immunohistochemically staining methods are the typically used techniques for the quantification of those proteins. As the costs for reagents and devices as well as the work schedule of these methods are immense it was the goal of our study to develop an easy and reliable method to quantify the concentration of specific proteins. We established a procedure to measure the relative concentration of proteins fixed on ROTI PVDF membranes via Western blot, calculating the relative protein concentration in dependency to the grey scale index of the normalized and digitalized pictures of the bands on the blots.
    Keywords: Quantification ; Protein ; Grey Scale ; Western Blot ; Hsp70 Superfamily ; Immune Detection ; Public Health
    ISSN: 0379-0738
    E-ISSN: 1872-6283
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  • 10
    Language: English
    In: International Journal of Health Care Quality Assurance, 11 February 2019, Vol.32(1), pp.281-295
    Description: Purpose The purpose of this paper is to examine the competencies that US healthcare organizations require for quality and performance improvement positions. Design/methodology/approach A US healthcare improvement job posting content analysis was conducted using the HQ Essentials competency framework. Findings The HQ essentials competencies most desired for improvement positions include project management, training, data analysis and applied performance improvement methods. Competency requirements varied somewhat by job focus area: performance, quality, or process improvement, and Lean and Six Sigma. Practical implications Healthcare leaders may use the author’s results to understand what competencies may be required for various improvement roles and to identify any gaps in required skills and knowledge areas that may need to be addressed. Educators and policy-makers should consider how these competencies align with employers’ needs and what resources or professional development may be needed to address gaps. Originality/value This is the first healthcare improvement competencies analysis based on job postings.
    Keywords: Performance Measurement ; Six Sigma ; Competencies ; Lean ; Process Efficiency ; Quality Improvement ; Performance Improvement ; Organizational Performance ; Medicine
    ISSN: 0952-6862
    E-ISSN: 1758-6542
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