Enhanced Antifibrinolytic Efficacy of a Plasmin-Specific Kunitz-Inhibitor (60-Residue Y11T/L17R with C-Terminal IEK) of Human Tissue Factor Pathway Inhibitor Type-2 Domain1.

Current antifibrinolytic agents reduce blood loss by inhibiting plasmin active sites (e.g., aprotinin) or by preventing plasminogen/tissue plasminogen activator (tPA) binding to fibrin clots (e.

[ICHI-International Classification of Health Interventions : A balancing act between the demands of statistics and reimbursement].

Medical classifications systematize medical concepts (e. g. diagnoses, procedures).

Systematic study of constitutive cyclooxygenase-2 expression: Role of NF-κB and NFAT transcriptional pathways.

Cyclooxygenase-2 (COX-2) is an inducible enzyme that drives inflammation and is the therapeutic target for widely used nonsteroidal antiinflammatory drugs (NSAIDs). However, COX-2 is also constitutively expressed, in the absence of overt inflammation, with a specific tissue distribution that includes the kidney, gastrointestinal tract, brain, and thymus. Constitutive COX-2 expression is therapeutically important because NSAIDs cause cardiovascular and renal side effects in otherwise healthy individuals.

Hepatocyte Heparan Sulfate Is Required for Adeno-Associated Virus 2 but Dispensable for Adenovirus 5 Liver Transduction In Vivo.

Unlabelled: Adeno-associated virus 2 (AAV2) and adenovirus 5 (Ad5) are promising gene therapy vectors. Both display liver tropism and are currently thought to enter hepatocytes in vivo through cell surface heparan sulfate proteoglycans (HSPGs). To test directly this hypothesis, we created mice that lack Ext1, an enzyme required for heparan sulfate biosynthesis, in hepatocytes.

Platelets contain tissue factor pathway inhibitor-2 derived from megakaryocytes and inhibits fibrinolysis.

Tissue factor pathway inhibitor-2 (TFPI-2) is a homologue of TFPI-1 and contains three Kunitz-type domains and a basic C terminus region. The N-terminal domain of TFPI-2 is the only inhibitory domain, and it inhibits plasma kallikrein, factor XIa, and plasmin. However, plasma TFPI-2 levels are negligible (≤20 pM) in the context of influencing clotting or fibrinolysis.

Reversible suppression of cyclooxygenase 2 (COX-2) expression in vivo by inducible RNA interference.

Prostaglandin-endoperoxide synthase 2 (PTGS2), also known as cyclooxygenase 2 (COX-2), plays a critical role in many normal physiological functions and modulates a variety of pathological conditions. The ability to turn endogenous COX-2 on and off in a reversible fashion, at specific times and in specific cell types, would be a powerful tool in determining its role in many contexts. To achieve this goal, we took advantage of a recently developed RNA interference system in mice.

Reduced T-dependent humoral immunity in CD20-deficient mice.

CD20 is a tetraspanning membrane protein expressed on B lymphocytes. CD20 deficiency in both mice and humans has recently been shown to have deleterious effects on Ab responses to T-independent Ags; however, no effect on T-dependent immunity has been reported. In this study, we used a Cd20⁻/⁻ mouse line to evaluate Ab responses to adeno-associated virus and SRBCs.

LC-MS/MS confirms that COX-1 drives vascular prostacyclin whilst gene expression pattern reveals non-vascular sites of COX-2 expression.

There are two schools of thought regarding the cyclooxygenase (COX) isoform active in the vasculature. Using urinary prostacyclin markers some groups have proposed that vascular COX-2 drives prostacyclin release. In contrast, we and others have found that COX-1, not COX-2, is responsible for vascular prostacyclin production.

Differential COX-2 induction by viral and bacterial PAMPs: Consequences for cytokine and interferon responses and implications for anti-viral COX-2 directed therapies.

Cyclooxygenase 2 (COX)-2 is induced by bacterial and viral infections and has complex, poorly understood roles in anti-pathogen immunity. Here, we use a knock-in luciferase reporter model to image Cox2 expression across a range of tissues in mice following treatment with the either the prototypical bacterial pathogen-associated molecular pattern (PAMP), LPS, which activates Toll-like receptor (TLR)4, or with poly(I:C), a viral PAMP, which activates TLR3. LPS induced Cox2 expression in all tissues examined.

Development of a new international classification of health interventions based on an ontology framework.

: The WHO International Classification of Diseases is used in many national applications to plan, manage and fund through case mix health care systems and allows international comparisons of the performance of these systems. There is no such measuring tool for health interventions or procedures. To fulfil this requirement the WHO-FIC Network recommended in 2006 to develop an International Classification of Health Interventions (ICHI).

Checking coding completeness by mining discharge summaries.

Incomplete coding is a known problem in hospital information systems. In order to detect non-coded secondary diseases we developed a text classification system which scans discharge summaries for drug names. Using a drug knowledge base in which drug names are linked to sets of ICD-10 codes, the system selects those documents in which a drug name occurs that is not justified by any ICD-10 code within the corresponding record in the patient database.

Differential effects of murine and human factor X on adenovirus transduction via cell-surface heparan sulfate.

Serum coagulation factor X (FX) is proposed to play a major role in adenovirus tropism, promoting transduction by bridging the virus to cell-surface heparan sulfate proteoglycans (HSPGs). Both murine FX and human FX increased transduction by Ad.CMVfLuc, an adenovirus vector, in murine hepatocyte-like cells and human hepatocarcinoma cells.

Health care procedures.

Background: ICD is used for coding medical diagnoses across the world, but there is no globally accepted coding system for health care procedures. The need for the introduction of a common international medical procedure classification has been addressed by the Australian NCCH, which proposed the International Classification of Health Interventions (ICHI) as the basis of an international procedure classification. In parallel, the French multiaxial Classification Commune des Actes Médicaux (CCAM) has been established.

The influence of innate and pre-existing immunity on adenovirus therapy.

Recombinant adenovirus serotype 5 (Ad5) vectors have been studied extensively in preclinical gene therapy models and in a range of clinical trials. However, innate immune responses to adenovirus vectors limit effectiveness of Ad5 based therapies. Moreover, extensive pre-existing Ad5 immunity in human populations will likely limit the clinical utility of adenovirus vectors, unless methods to circumvent neutralizing antibodies that bind virus and block target cell transduction can be developed.

Antiviral antibodies target adenovirus to phagolysosomes and amplify the innate immune response.

Adenovirus is a nonenveloped dsDNA virus that activates intracellular innate immune pathways. In vivo, adenovirus-immunized mice displayed an enhanced innate immune response and diminished virus-mediated gene delivery following challenge with the adenovirus vector AdLacZ suggesting that antiviral Abs modulate viral interactions with innate immune cells. Under naive serum conditions in vitro, adenovirus binding and internalization in macrophages and the subsequent activation of innate immune mechanisms were inefficient.

Immunity to adeno-associated virus vectors in animals and humans: a continued challenge.

Recombinant vectors based on adeno-associated virus (AAV) have been shown to stably express many genes in vivo without mounting immune responses to vectors or transgenes. Thus, AAV vectors have rapidly become the reagents of choice for therapeutic gene transfer. Yet one of the first translations of AAV gene therapy into humans unexpectedly resulted in only short-term expression of the therapeutic gene accompanied by transient but significant toxicity.

The inflammasome recognizes cytosolic microbial and host DNA and triggers an innate immune response.

The innate immune system recognizes nucleic acids during infection and tissue damage. Whereas viral RNA is detected by endosomal toll-like receptors (TLR3, TLR7, TLR8) and cytoplasmic RIG-I and MDA5, endosomal TLR9 and cytoplasmic DAI bind DNA, resulting in the activation of nuclear factor-kappaB and interferon regulatory factor transcription factors. However, viruses also trigger pro-inflammatory responses, which remain poorly defined.

Complement is an essential component of the immune response to adeno-associated virus vectors.

Adeno-associated virus (AAV) vectors are associated with relatively mild host immune responses in vivo. Although AAV induces very weak innate immune responses, neutralizing antibodies against the vector capsid and transgene still occur. To understand further the basis of the antiviral immune response to AAV vectors, studies were performed to characterize AAV interactions with macrophages.

[The French Common Classification of Procedures CCAM. An option for Germany].

A working group of the National Board for Classification in Health Care at the Federal Ministry for Health (KKG) concerned itself between 1996 and 2003 with the topic of a possible follow-up procedure classification system for the actually used "Operation and Procedure Coding System" (OPS-301), which was introduced to Germany in the mid 1990's. In the context of a feasibility study the American "Procedure Coding system" (PCS) and the French "Classification commune des actes médicaux "(CCAM) were examined and evaluated as possible follow-up classifications. In this study it could be shown that the CCAM has great advantages over the pure multiaxial PCS.

Comparison of ICHI and CCAM basic coding system.

While diagnoses are coded by ICD across the world, there is no universally accepted coding system for procedures. In many countries there exists not even a local classification for medical procedures. As a possible solution the International Classification of Health Interventions (ICHI) has been proposed as a common denominator for an international procedure classification.

Neutrophils interact with adenovirus vectors via Fc receptors and complement receptor 1.

Neutrophils are effectors of the innate immune response to adenovirus vectors. Following the systemic administration of Cy2-labeled AdLuc in mice, flow cytometry and PCR analysis of liver leukocytes revealed that 25% of recruited neutrophils interacted with adenovirus vectors. In vitro, flow cytometry of human neutrophils incubated with Cy2-labeled AdLuc also demonstrated a significant interaction with adenovirus vectors.

Immune responses to adeno-associated virus vectors.

One of the biggest challenges in optimizing viral vectors for gene therapy relates to the immune response of the host. Adeno-associated virus (AAV) vectors are associated with low immunogenicity and toxicity, resulting in vector persistence and long-term transgene expression. The inability of AAV vectors to efficiently transduce or activate antigen presenting cells (APCs) may account for their decreased immunogenicity.

Helper-dependent adenovirus vectors elicit intact innate but attenuated adaptive host immune responses in vivo.

Helper-dependent adenovirus (HD-Ad) vectors with all adenoviral genes deleted mediate very long-term expression of therapeutic transgenes in a variety of animal models of disease. These vectors are associated with reduced toxicity and improved safety relative to traditional early region 1 deletion first-generation Ad (FG-Ad) vectors. Many studies have clearly demonstrated that FG-Ad vectors induce innate and adaptive immune responses in vivo; however, a comprehensive analysis of host immune responses to HD-Ad vectors has not yet been performed.

[Electronic distribution of patient records and images through PACS: implementation of data protection at the University Hospital in Freiburg].

In large installations of a Picture Archiving and Communication System (PACS), data protection becomes an issue. Above all, not every employee should have access to all data stored in the system. At the Freiburg University Hospital, we developed a concept of dynamic assignment of authorization, assuring that every physician is authorized to the data of all patients under his care.

The role of capsid-endothelial interactions in the innate immune response to adenovirus vectors.

Adenovirus (Ad) vectors can produce inflammatory responses at high doses. Intravenous administration of an Ad vector expressing green fluorescent protein (AdGFP) to naive mice induced a biphasic pattern of liver cytokine/chemokine gene expression over 7 days. Tumor necrosis factor alpha (TNF-alpha), macrophage inflammatory protein 2 (MIP-2), and interferon gamma-inducible protein 10 (IP-10) genes were upregulated, with two distinct peaks of mRNA expression occurring at 6 hr and 5 days.