Rapid selection of specific MAP kinase-binders from designed ankyrin repeat protein libraries

We describe here the rapid selection of specific MAP-kinase binders from a combinatorial library of designed ankyrin repeat proteins (DARPins). A combined in vitro/in vivo selection approach, based on ribosome display and the protein fragment complementation assay (PCA), yielded a large number of different binders that are fully functional in the cellular cytoplasm. Ribosome-display selection pools of four successive selection rounds were examined to monitor the enrichment of JNK2-specific DARPins. Surprisingly, only one round of ribosome display with subsequent PCA selection of this pool was necessary to isolate a first specific binder with micromolar affinity. After only two rounds of ribosome-display selection followed by PCA, virtually all DARPins showed JNK2-specific binding, with affinities in the low nanomolar range. The enrichment factor of ribosome display thus approaches 105 per round. In a second set of experiments, similar results were obtained with the kinases JNK1 and p38 as targets. Again, almost all investigated DARPins obtained after two rounds of ribosome display showed specific binding to the targets used, JNK1 or p38. In all three selection experiments the identified DARPins possess very high specificity for the target kinase. Taken together, the combination of ribosome display and PCA selections allowed the identification of large pools of binders at unparalleled speed. Furthermore, DARPins are applicable in intracellular selections and immunoprecipitations from the extract of eukaryotic cell

Data on the Number of Somites Compared with Age in the White Rat

Author Institution: Department of Anatomy, University of California and Ohio State Universit

Oxygen reduction at soft interfaces catalyzed by in situ-generated reduced graphene oxide

peer-reviewedFace to face: Flakes of reduced graphene oxide, synthesized in situ at the liquid/liquid interface from a graphene‐oxide precursor, are capable of catalyzing the biphasic reduction of protons to hydrogen peroxide in the presence of molecular oxygen and an organic solubilized electron donor. This offers a new perspective for the bulk production of a green oxidant through biphasic electrolysisACCEPTEDpeer-reviewe

Inkjet Printed Nanohydrogel Coated Carbon Nanotubes Electrodes For Matrix Independent Sensing

Polyacrylamide (PA) based hydrogels are used in several applications including polyacrylamide gel electrophoresis and sensing devices. Homogeneous and compact PA films can be prepared based on chemical or photopolymerization processes. However, the accurate and reproducible coating of substrates with nanohydrogel patterns is challenging due to the in situ polymerization and deposition requirements. Herein, we report an inkjet printing (IJP) concept with simultaneously performed UV photopolymerization of a specifically prepared acrylamide/N,N′-methylenebis(acrylamide) containing ink. A prepolymerization step of the hydrogel precursor molecules was implemented in the ink formulation protocol to adjust the viscosity of the ink and to enhance the rate of polymerization during printing. After the optimization of the printing parameters, a nanometer thin PA hydrogel coating with well distributed nanopores was achieved on top of a stand-alone carbon nanotubes (CNTs) pattern. Batches of fully inkjet printed PA/CNT modified electrodes were prepared that showed outstanding improvements for the electrochemical detection of antioxidants in complex matrices such as untreated orange juice and red wine samples thanks to the properties of the PA coating

Rapid selection of specific MAP kinase-binders from designed ankyrin repeat protein libraries

We describe here the rapid selection of specific MAP-kinase binders from a combinatorial library of designed ankyrin repeat proteins (DARPins). A combined in vitro/in vivo selection approach, based on ribosome display and the protein fragment complementation assay (PCA), yielded a large number of different binders that are fully functional in the cellular cytoplasm. Ribosome-display selection pools of four successive selection rounds were examined to monitor the enrichment of JNK2-specific DARPins. Surprisingly, only one round of ribosome display with subsequent PCA selection of this pool was necessary to isolate a first specific binder with micromolar affinity. After only two rounds of ribosome-display selection followed by PCA, virtually all DARPins showed JNK2-specific binding, with affinities in the low nanomolar range. The enrichment factor of ribosome display thus approaches 105 per round. In a second set of experiments, similar results were obtained with the kinases JNK1 and p38 as targets. Again, almost all investigated DARPins obtained after two rounds of ribosome display showed specific binding to the targets used, JNK1 or p38. In all three selection experiments the identified DARPins possess very high specificity for the target kinase. Taken together, the combination of ribosome display and PCA selections allowed the identification of large pools of binders at unparalleled speed. Furthermore, DARPins are applicable in intracellular selections and immunoprecipitations from the extract of eukaryotic cell

Metal on metal hip resurfacing versus uncemented custom total hip replacement - early results

<p>Abstract</p> <p>Introduction</p> <p>There is no current consensus on the most appropriate prosthesis for treating symptomatic osteoarthritis (OA) of the hip in young, active patients. Modern metal on metal hip resurfacing arthroplasty (HR) has gained popularity as it is theoretically more stable, bone conserving and easier to revise than total hip arthroplasty. Early results of metal on metal resurfacing have been encouraging. We have compared two well matched cohorts of patients with regard to function, pain relief and patient satisfaction.</p> <p>Methods</p> <p>This prospective study compares 2 cohorts of young, active patients treated with hip resurfacing (137 patients, 141 hips) and custom uncemented (CADCAM) stems (134 patients, 141 hips). All procedures were performed by a single surgeon. Outcome measures included Oxford, WOMAC and Harris hip scores as well as an activity score. Statistical analysis was performed using the unpaired student's t-test.</p> <p>Results</p> <p>One hundred and thirty four and 137 patients were included in the hip replacement and resurfacing groups respectively. The mean age of these patients was 54.6 years. The mean duration of follow up for the hip resurfacing group was 19.2 months compared to 13.4 months for the total hip replacement group.</p> <p>Pre operative oxford, Harris and WOMAC scores in the THA group were 41.1, 46.4 and 50.9 respectively while the post operative scores were 14.8, 95.8 and 5.0. In the HR group, pre- operative scores were 37.0, 54.1 and 45.9 respectively compared to 15.0, 96.8 and 6.1 post operatively. The degree of improvement was similar in both groups.</p> <p>Conclusion</p> <p>There was no significant clinical difference between the patients treated with hip resurfacing and total hip arthroplasty in the short term.</p

Inkjet Printing Meets Electrochemical Energy Conversion

Inkjet printing is a very powerful digital and mask-less microfabrication technique that has attracted the attention of several research groups working on electrochemical energy conversion concepts. In this short review, an overview is given about recent efforts to employ inkjet printing for the search of new electrocatalyst materials and for the preparation of catalyst layers for polymer electrolyte membrane fuel cell applications. Recent approaches of the Laboratory of Physical and Analytical Electrochemistry (LEPA) at the École Polytechnique Fédérale de Lausanne for the inkjet printing of catalyst layers and membrane electrode assemblies are presented and future energy research directions of LEPA based on inkjet printing in the new Energypolis campus in the Canton of Valais are summarized

Community-based care models for arterial hypertension management in non-pregnant adults in sub-Saharan Africa: a literature scoping review and framework for designing chronic services

BACKGROUND: Arterial hypertension (aHT) is the leading cardiovascular disease (CVD) risk factor in sub-Saharan Africa; it remains, however, underdiagnosed, and undertreated. Community-based care services could potentially expand access to aHT diagnosis and treatment in underserved communities. In this scoping review, we catalogued, described, and appraised community-based care models for aHT in sub-Saharan Africa, considering their acceptability, engagement in care and clinical outcomes. Additionally, we developed a framework to design and describe service delivery models for long-term aHT care. METHODS: We searched relevant references in Embase Elsevier, MEDLINE Ovid, CINAHL EBSCOhost and Scopus. Included studies described models where substantial care occurred outside a formal health facility and reported on acceptability, blood pressure (BP) control, engagement in care, or end-organ damage. We summarized the interventions' characteristics, effectiveness, and evaluated the quality of included studies. Considering the common integrating elements of aHT care services, we conceptualized a general framework to guide the design of service models for aHT. RESULTS: We identified 18,695 records, screened 4,954 and included twelve studies. Four types of aHT care models were identified: services provided at community pharmacies, out-of-facility, household services, and aHT treatment groups. Two studies reported on acceptability, eleven on BP control, ten on engagement in care and one on end-organ damage. Most studies reported significant reductions in BP values and improved access to comprehensive CVDs services through task-sharing. Major reported shortcomings included high attrition rates and their nature as parallel, non-integrated models of care. The overall quality of the studies was low, with high risk of bias, and most of the studies did not include comparisons with routine facility-based care. CONCLUSIONS: The overall quality of available evidence on community-based aHT care is low. Published models of care are very heterogeneous and available evidence is insufficient to recommend or refute further scale up in sub-Sahara Africa. We propose that future projects and studies implementing and assessing community-based models for aHT care are designed and described according to six building blocks: providers, target groups, components, location, time of service delivery, and their use of information systems