Extended-spectrum ß-lactamase (ESBL) detection directly from urine samples with the rapid isothermal amplification-based eazyplex® SuperBug CRE assay: Proof of concept.

A commercially available assay (eazyplex® SuperBug CRE) detecting the most common carbapenemase and ESBL types was evaluated directly on 50 urine samples. Eazyplex® correctly detected ESBL-encoding genes in all 30 urine samples with confirmed ESBL production (sensitivity 100%). Two specimens showed invalid and one specimen false-positive results (specificity 97.9%).

Identification of a novel 16S rRNA gene variant of Actinomyces funkei from six patients with purulent infections.

Little is known about the clinical significance and laboratory diagnosis of Actinomyces funkei. In this report we describe six clinical cases where A. funkei was isolated from purulent, polymicrobial infections. Conventional identification procedures were compared with molecular methods including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry technique. Analysis of the full 16S rRNA gene sequence of the six investigated strains revealed differences from the A. funkei type strain. DNA-DNA hybridization showed that the clinical strains represent a novel 16S rRNA gene variant within the species of A. funkei.

Corynebacterium tuberculostearicum: a potentially misidentified and multiresistant Corynebacterium species isolated from clinical specimens.

Corynebacterium tuberculostearicum is a lipophilic corynebacterium validly characterized in 2004. We provide clinical information on 18 patients from whom this organism was isolated. The majority of the patients were hospitalized and had a history of prolonged treatment with broad-spectrum antimicrobials. In 7 (38.9%) of the 18 cases, the isolates were found to be clinically relevant. The present report also includes detailed data on the biochemical and molecular identification of C. tuberculostearicum, as well as its identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Our data demonstrate that routine biochemical tests do not provide reliable identification of C. tuberculostearicum. MALDI-TOF MS represents a helpful tool for the identification of this species, since all of the strains matched C. tuberculostearicum as the first choice and 58.3% (7/12) of the strains processed with the full extraction protocol generated scores of >2.000. Nevertheless, partial 16S rRNA gene sequencing still represents the gold standard for the identification of this species. Due to the challenging identification of C. tuberculostearicum, we presume that this organism is often misidentified and its clinical relevance is underestimated. The antimicrobial susceptibility profile of C. tuberculostearicum presented here reveals that 14 (87.5%) of the 16 strains analyzed exhibited multidrug resistance.

Smac mimetics: implications for enhancement of targeted therapies in leukemia.

Drug resistance is a growing concern with clinical use of tyrosine kinase inhibitors. Utilizing in vitro models of intrinsic drug resistance and stromal-mediated chemoresistance, as well as functional mouse models of progressive and residual disease, we attempted to develop a potential therapeutic approach designed to suppress leukemia recurrence following treatment with selective kinase inhibitors. The novel IAP inhibitor, LCL161, [corrected] was observed to potentiate the effects of tyrosine kinase inhibition against leukemic disease both in the absence and presence of a stromal-protected [corrected] environment. LCL161 enhanced the proapoptotic effects of nilotinib and PKC412, against leukemic disease in vitro and potentiated the activity of both kinase inhibitors against leukemic disease in vivo. In addition, LCL161 synergized in vivo with nilotinib to reduce leukemia burden significantly below the baseline level suppression exhibited by a moderate-to-high dose of nilotinib. Finally, LCL161 displayed antiproliferative effects against cells characterized by intrinsic resistance to tyrosine kinase inhibitors as a result of expression of point mutations in the protein targets of drug inhibition. These results support the idea of using IAP inhibitors in conjunction with targeted tyrosine kinase inhibition to override drug resistance and suppress or eradicate residual disease.

Translating research into practice: a German sickness fund supporting patient participation.

OBJECTIVE: To demonstrate how a German sickness fund like the Techniker Krankenkasse (TK) tries to translate evidence from research in the field of shared decision-making (SDM) into practice. METHODS: This article outlines three different initiatives of the TK, supporting patient participation in every day life: information tools, continuous interactive case management, and web-based interactive patient dialogue. These three activities are supposed to show a stepwise development- and introduction-process which is designed to include patient feedback at all stages of progress. Wherever available, empirical data is added to the description of the initiatives. This data contains information about how many people use a special service offered and how useful or satisfying the service is in the users' eyes. RESULTS: The examples given show that a single sickness fund is able to support patient participation with different kinds of initiatives. CONCLUSION: Implementation of SDM will be beneficial for the statutory health insurance and will have a positive influence on political and professional decision processes. PRACTICE IMPLICATIONS: The TK will continue and intensify its engagement for SDM in the future.

Scientific and standardization committee communications: classification and nomenclature of disintegrins isolated from snake venoms.

Viper venoms from the Elapidae, Hydrophiidae, Atractaspididae, Viperidae and Colubridae families contain at least 25 separate classes of biologically active compounds, including enzymes and non-enzymatic molecules. Nomenclature standardization of these by structure and function has been reported for prothrombin activators. No such agreement has been made for the nomenclature and classification of disintegrins. At the 51st Annual SSC Meeting in Sydney Australia (August 2005), we reported the variety of names given to the 78 disintegrins then known, and proposed a nomenclature standardization method for the naming of future members in this protein family. The summary of these recommendations will provide guidance as disintegrin discoveries move beyond high pressure liquid chromatography (HPLC) separation methods to mass spectrometry, proteomics and cloned cDNA.

Trophic transfer of polychlorinated biphenyls in great blue heron (Ardea herodias) at Crab Orchard National Wildlife Refuge, Illinois, United States.

In this study, polychlorinated biphenyl (PCB) concentrations were measured in great blue heron (GBHE) (Ardea herodias) chicks and eggs at Crab Orchard National Wildlife Refuge (CONWR) in southern Illinois. In addition, biomagnification factors (BMFs) from gizzard shad (Dorosoma cepedianum) and their effects on reproductive effort were examined. Total PCBs (SigmaPCBs) in chicks and shad were greater at the east end of Crab Orchard Lake (i.e., near the site of contamination) than the west end, but chick concentrations (4.1 to 10.1 mg/kg lipid weight) were lower than those typically associated with adverse effects. Chick BMFs based on shad from diet samples were greater than those based on shad collected from the lake. Furthermore, the two shad sources had dissimilar dioxin-like congener patterns and SigmaPCBs, suggesting that there was variation in PCB load and composition and that the more contaminated shad were a small proportion of the actual heron chick diet. The number of eggs laid per nest was similar between colonies, suggesting no observable population level effects. Further study may be necessary to evaluate long-term effects on GBHEs at CONWR.

[Contribution of leflunomide to the cost effectiveness of sequential DMARD therapy of rheumatoid arthritis in Germany]. / Wirtschaftlichkeit von Leflunomid bei sequentieller Basistherapie der rheumatoiden Arthritis in Deutschland.

Since November 1999, leflunomide (LEF), an innovative disease-modifying antirheumatic drug (DMARD), is available in Germany for treatment of rheumatoid arthritis (RA). LEF slows radiographic disease progression and improves functional capacity as well as healthrelated quality of life of RA patients. Resources for health care of the patients are limited in Germany as in all other countries. The purpose of the analysis therefore was to compare the cost effectiveness of the following alternatives: LEF in sequential monotherapy with other DMARDs versus sequential monotherapy of other DMARDs. The target variables of this cost-effectiveness comparison were additional direct costs per ACR20-response year (ACR20RY) gained and per quality-adjusted life year (QALY) gained, respectively, each after three years of treatment. The cost-effectiveness comparison was carried out using a modeling study after secondary analysis of relevant data. Oral methotrexate (MTX), sulphasalazine (SSZ), antimalarials (CQ/HCQ), intramuscular gold (IMG), and azathioprine (AZA) were selected as "other" DMARDs representing the current status of sequential monotherapy. Based on health care regulation in Germany-Guidelines on the Prescription of Drugs amended by the Federal Commission of Medical Practitioners and Health Insurance Funds on 10 December 1999-LEF was exclusively considered second within a DMARD sequence. Direct costs were given by outpatient and inpatient treatment, long-term care, and rehabilitation treatment. Prices relate to the period of 1998 to 2001 and were converted to Euro (euro), according to the official exchange rate of 1 euro = 1.95583 DM (1 euro approximately 0.90 US dollars; 2001 values). The comparative cost-effectiveness analysis covered a treatment period of more than one year. To estimate the net present value of future costs and effectiveness, a discount rate of 5% per year was applied. In the case of DMARD-naïve patients with RA, the sequence MTX, LEF, SSZ, IMG, AZA, CQ/HCQ was the most cost effective with direct costs of 7297 euro per ACR20RY and 6499 euro per QALY. In order to estimate the consequences of introducing LEF into the prescribing practice in Germany, the distribution of RA patients by individual DMARD in rheumatological care in 1998 was considered. This distribution was taken from the National Database of the German Collaborative Arthritis Centres. Though the sequences comprising LEF incurred 3% higher direct costs, they led to a higher effectiveness of 6% and 3% in the case of ACR20RYs and QALYs, respectively. Choosing sequences comprising LEF, there were additional direct costs of 5004 euro per ACR20RY gained and 8301 euro per QALY gained, as compared to the corresponding sequences without LEF. In comprehensive sensitivity analyses, the robustness of the model and its results was shown. The contribution of LEF to the cost effectiveness of sequential DMARD therapy is obvious. The modeling study revealed advantages for the patients and the cost carriers. Though there were initially higher medication costs of the sequences comprising LEF, these costs were nearly compensated to remaining excess costs of just 3% after three years. This was caused by cost savings in other sectors of the health care system due to the higher effectiveness of the sequences comprising LEF.

New insights on disintegrin-receptor interactions: eristostatin and melanoma cells.

Viper venom disintegrins have been used frequently to study the cellular receptors which characterize various types of cells, including platelets, endothelial cells and cancer cells. While the majority of such analyses have pointed to involvement of integrin receptors alphavbeta3, alpha5beta1 or alphaIIbbeta3, this may not always be so. Eristostatin, from Eristocophis macmahoni, is a potent inhibitor of ADP-induced platelet aggregation as well as of human and murine melanoma metastases in mouse model systems. This disintegrin requires an RGDW motif, as well as an intact C-terminus, in order to interact with both platelets and four different types of melanoma cells. Eristostatin causes nonmetastatic SBc12 melanoma cells to show higher susceptibility to specific killing by NK-like TALL-104 cells. While it is known that eristostatin binds to alphaIIbbeta3 on platelets, the receptor with which eristostatin binds to the melanoma cells has not yet been identified.

Synthesis of the angiotensin converting enzyme inhibitor (-)-A58365A via an isomünchnone cycloaddition reaction.

[formula: see text] The angiotensin converting enzyme inhibitor (-)-A58365A (1) was synthesized by a process based on the [3 + 2]-cycloaddition reaction of a phenylsulfonyl-substituted isomünchnone intermediate. The starting material for this process was prepared from L-pyroglutamic acid and involved using a diazo-phenylsulfonyl-substituted pyrrolidine imide. Treatment of the diazoimide with Rh2(OAc)4 in the presence of methyl vinyl ketone afforded a 3-hydroxy-2-pyridone derivative which was subsequently converted to the ACE inhibitor in six additional steps.

[A concept for a clearing procedure for guidelines in Germany]. / Konzept eines Clearingverfahrens für Leitlinien in Deutschland.

Guidelines are efficient tools for the maintenance and improvement of the quality of care and contribute to the increase of the efficacy of care. However, the quality of guidelines has not been investigated or improved much in Germany. This is especially true for the formal assessment of guidelines regarding the collection and evaluation of evidence, their feasibility and their effects on health economy. There is an internationally recognized method available to examine and evaluate current and new guidelines with respect to these three categories. This paper proposes a clearing procedure for guidelines in Germany to systematically improve the quality of guidelines and to offer authors as well as users of guidelines valuable informations.

Evaluation of an orientation system for newly employed registered nurses.

In this article, the authors discussed the successful evaluation of an orientation system for newly employed registered nurses in a large teaching hospital using the IOP model. This methodology can be successfully applied to any educational program that is consolidated into an organization's goals. Although not well examined, orientation has been reported to be costly (Bethel, 1992; del Bueno, Weeks, Brown-Stewart, 1987). The system presently used at this hospital uses at least 1 week of a nurse educator's time, 3-10 weeks for a newly employed registered nurse, and 3-10 weeks for a preceptor RN. Such an investment of personnel resources mandates examination of the processes and outcomes of the program to ensure newly employed RNs become competent practitioners as efficiently as possible. The use of the IOP model particularly was useful in examining a complex orientation system in a multicentered hospital. Use of this systematic program evaluation separated the overall orientation process into workable components. Tools, such as the algorithm, allowed for easy visualization and comprehension of the process steps. This was indispensable because of the number and scope of people involved in the orientation program. The evaluation process was impartial and focused on the program steps, not on the individuals. Because of this impartiality, people were able to gather and work cohesively to improve the overall program. Use of the IOP model assisted the nurse educators in determining that PBDS was not achieving the goal of identifying individual learning needs. Rather, PBDS was a useful tool in establishing baseline competency of newly employed RNs. The system clearly identified those individuals who had above average knowledge bases and those individuals who had more learning needs. For those with more learning needs, PBDS provides a starting point for planning a structured orientation. Thus, a Phase II PBDS assessment could be used as a more unit-specific assessment to validate whether the RN has achieved the orientation objectives. Although the IOP model is not a strict research methodology, it is appropriate for examination of a program as fluid and ongoing as this. Finally, ongoing run charts or statistical trends will assist the nurse educators in monitoring the quality and effectiveness of the orientation program.

Complete amino acid sequence of ovine miniplasminogen.

The complete amino acid sequence of ovine miniplasminogen (M(r) 37,662, 343 residues) was determined with the aid of fragments obtained by cleavage with 2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine and clostripain. The fragments were aligned with overlapping sequences and sequence comparison with miniplasminogens of other species. Sequence comparison with other species (human, bovine, porcine, equine and canine) gave an overall identity of 63% and a similarity of 83%. The dendrogram of the alignment indicates that ovine miniplasminogen has the closest relationship with the bovine (87% identity) and the most distant with the equine (77% identity) species. The close relationship is indicative for the presence of the same structural and functional domains as in the other species. Sequence comparison of different miniplasminogens showed that positions 49 (Arg), 83 (Arg) and 161 (Ser) in the light chain of the plasmin molecule may play a role in the interaction between plasminogen and streptokinase.

Complete amino acid sequence of equine miniplasminogen.

The complete amino acid sequence of equine miniplasminogen (Mr 37,132, 338 residues) was determined with the aid of fragments obtained by cleavage with 2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine, cyanogen bromide or clostripain. The fragments were aligned with overlapping sequences. Sequence comparison with other species gave identities in the range of 76% (bovine) and 81% (canine), indicating the presence of the same structural and functional domains as in the other species. Sequence comparison of different miniplasminogens showed that positions 49 (Arg), 83 (Arg) and 161 (Ser) may play a role in the interaction between plasminogen and streptokinase.