Aspens Among the Pines

Grove of aspens among pine trees in Twin Lakes, Colorado

‘Maybe we can turn the tide’ : an explanatory mixed-methods study to understand how knowledge brokers mobilise health evidence in low- and middle-income countries

Background: Little is known about how knowledge brokers (KBs) operate in low- and middle-income countries (LMICs) to translate evidence for health policy and practice. These intermediaries facilitate relationships between evidence producers and users to address public health issues. Aims and objectives: To increase understanding, a mixed-methods study collected data from KBs who had acted on evidence from the 2015 Global Maternal Newborn Health Conference in Mexico. Methods: Of the 1000 in-person participants, 252 plus 72 online participants (n=324) from 56 countries completed an online survey, and 20 participants from 15 countries were interviewed. Thematic analysis and application of knowledge translation (KT) theory explored factors influencing KB actions leading to evidence uptake. Descriptive statistics of respondent characteristics were used for cross-case comparison. Findings: Results suggest factors supporting the KB role in evidence uptake, which include active relationships with evidence users through embedded KB roles, targeted and tailored evidence communication to fit the context, user receptiveness to evidence from a similar country setting, adaptability in the KB role, and action orientation of KBs. Discussion and conclusions: Initiatives to increase evidence uptake in LMICs should work to establish supportive structures for embedded KT, identify processes for ongoing cross-country learning, and strengthen KBs already showing effectiveness in their roles

Xenopus Development from Late Gastrulation to Feeding Tadpole in Simulated Microgravity

Microgravity (microG) is known to influence cytoskeletal structure, but its effects on cell migration are not well understood. To examine the effects of altered gravity on neural crest cell (NCC) migration, we inserted Xenopus laevis embryos into two separate microG-simulating slow turning lateral vessels (STLVs) just before neurulation (stage 11-12), and exposed them until feeding stage (stage 45), when the jaws and branchial apparatus are fully functional. To evaluate apparatus-related artifacts, we used two different STLVs and a vibration control as well as a stationary control vessel. Larval growth, pattern of NCC-derived cartilage formation, and incidence of malformations were analyzed using immunolocalization and wholemount staining of cartilage with Alcian blue. Interestingly, the two STLVs often yielded different or conflicting results. Many differences, such as increased cartilage size, attenuated Hoxa2 expression, and increased cell division, may be attributed mainly to vibration of the rotating vessels. However, tadpoles that developed in simulated microgravity (both STLVs, but not the vibration control) showed significantly more skeletal abnormalities, with stronger effects on cartilages derived from NCCs than those derived mainly from mesoderm. We conclude that migrating NCCs of Xenopus are sensitive to the altered gravitational environment of STLVs, and that studies relying on bioreactors to simulate microgravity also need to take variation in apparatus into account

A Three Monoclonal Antibody Combination Potently Neutralizes Multiple Botulinum Neurotoxin Serotype E Subtypes.

Human botulism is most commonly caused by botulinum neurotoxin (BoNT) serotypes A, B, and E. For this work, we sought to develop a human monoclonal antibody (mAb)-based antitoxin capable of binding and neutralizing multiple subtypes of BoNT/E. Libraries of yeast-displayed single chain Fv (scFv) antibodies were created from the heavy and light chain variable region genes of humans immunized with pentavalent-toxoid- and BoNT/E-binding scFv isolated by Fluorescence-Activated Cell Sorting (FACS). A total of 10 scFv were isolated that bound one or more BoNT/E subtypes with nanomolar-level equilibrium dissociation constants (KD). By diversifying the V-regions of the lead mAbs and selecting for cross-reactivity, we generated three scFv that bound all four BoNT/E subtypes tested at three non-overlapping epitopes. The scFvs were converted to IgG that had KD values for the different BoNT/E subtypes ranging from 9.7 nM to 2.28 pM. An equimolar combination of the three mAbs was able to potently neutralize BoNT/E1, BoNT/E3, and BoNT/E4 in a mouse neutralization assay. The mAbs have potential utility as therapeutics and as diagnostics capable of recognizing multiple BoNT/E subtypes. A derivative of the three-antibody combination (NTM-1633) is in pre-clinical development with an investigational new drug (IND) application filing expected in 2018

Airborne fine-resolution UHF radar: an approach to the study of englacial reflections, firn compaction and ice attenuation rates

This is the published version. Copyright 2015 International Glaciological SocietyWe have built and operated an ultra-wideband UHF pulsed-chirp radar for measuring firn stratigraphy from airborne platforms over the ice sheets of Greenland and West Antarctica. Our analysis found a wide range of capabilities, including imaging of post firn–ice transition horizons and sounding of shallow glaciers and ice shelves. Imaging of horizons to depths exceeding 600 m was possible in the colder interior regions of the ice sheet, where scattering from the ice surface and inclusions was minimal. The radar's high sensitivity and large dynamic range point to loss tangent variations as the dominant mechanism for these englacial reflective horizons. The radar is capable of mapping interfaces with reflection coefficients as low as –80 dB near the firn–ice transition and as low as –64 dB at depths of 600 m. We found that firn horizon reflectivity strongly mirrored density variance, a result of the near-unity interfacial transmission coefficients. Zones with differing compaction mechanisms were also apparent in the data. We were able to sound many ice shelves and areas of shallow ice. We estimated ice attenuation rates for a few locations, and our attenuation estimates for the Ross Ice Shelf, West Antarctica, appear to agree well with earlier reported results

A Rab4-like GTPase in Dictyostelium discoideum colocalizes with V-H(+)-ATPases in reticular membranes of the contractile vacuole complex and in lysosomes

In the course of screening a cDNA library for ras-related Dictyostelium discoideum genes, we cloned a 0.7 kb cDNA (rabD) encoding a putative protein that was 70% identical at the amino acid level to human Rab4. Rab4 is a small M(r) GTPase, which belongs to the Ras superfamily and functions to regulate endocytosis in mammalian cells. Southern blot analysis indicated that the rabD cDNA was encoded by a single copy gene while Northern blot analysis revealed that the rabD gene was expressed at relatively constant levels during growth and differentiation. Affinity-purified antibodies were prepared against a RabD fusion protein expressed in bacteria; the antibodies recognized a single 23 kDa polypeptide on western blots of cell extracts. Density gradient fractionation revealed that the RabD antigen co-distributed primarily with buoyant membranes rich in vacuolar protons pumps (V-H(+)-ATPases) and, to a lesser extent, with lysosomes. This result was confirmed by examining cell lines expressing an epitope-tagged version of RabD. Magnetically purified early endocytic vesicles and post-lysosomal vacuoles reacted more weakly with anti-RabD antibodies than did lysosomes. Other organelles were negative for RabD. Double-label indirect immunofluorescence microscopy revealed that RabD and the 100 kDa V-H(+)-ATPase subunit colocalized in a fine reticular network throughout the cytoplasm. This network was reminiscent of spongiomes, the tubular elements of the contractile vacuole system. Immunoelectron microscopy confirmed the presence of RabD in lysosome fractions and in the membranes rich in V-H(+)-ATPase. We conclude that a Rab4-like GTPase in D. discoideum is principally associated with the spongiomes of contractile vacuole complex

EVects of phenytoin and carbamazepine on calcium transport in Caco-2 cells

Abstract Adverse eVects of anti-seizure/anti-epileptic medications on bone density have been observed and reported since the early 1960s. Phenytoin and carbamazepine are two commonly prescribed anti-epileptic drugs most frequently associated with osteomalacia including fractures, bone demineralization, and reduced bone formation. The mechanism by which anti-epileptic drugs induce bone loss is not fully explained. We hypothesized that anti-epileptic drugs may impair dietary calcium absorption in the intestine. Using Caco-2 cells, a model transport system for study of the function of the intestinal epithelium, we determined the eVects of several anti-epileptic drugs on intestinal epithelial calcium transport. In our system, phenytoin and carbamazepine dose-dependently inhibit active calcium transport from the apical to basolateral side of Caco-2 cells under physiologic calcium conditions. Vitamin D ameliorates the anti-epileptic drug-induced decrease in calcium permeability

Extraction and inhibition of enzymatic activity of botulinum neurotoxins /B1, /B2, /B3, /B4, and /B5 by a panel of monoclonal anti-BoNT/B antibodies

<p>Abstract</p> <p>Background</p> <p>Botulism is caused by botulinum neurotoxins (BoNTs), extremely toxic proteins which can induce respiratory failure leading to long-term intensive care or death. Treatment for botulism includes administration of antitoxins, which must be administered early in the course of the intoxication; therefore, rapid determination of human exposure to BoNT is an important public health goal. In previous work, our laboratory reported on Endopep-MS, a mass spectrometry-based activity method for detecting and differentiating BoNT/A, /B, /E, and /F in clinical samples. We also demonstrated that antibody-capture is effective for purification and concentration of BoNTs from complex matrices such as clinical samples. However, some antibodies inhibit or neutralize the enzymatic activity of BoNT, so the choice of antibody for toxin extraction is critical.</p> <p>Results</p> <p>In this work, we evaluated 24 anti-BoNT/B monoclonal antibodies (mAbs) for their ability to inhibit the <it>in vitro </it>activity of BoNT/B1, /B2, /B3, /B4, and /B5 and to extract those toxins. Among the mAbs, there were significant differences in ability to extract BoNT/B subtypes and inhibitory effect on BoNT catalytic activity. Some of the mAbs tested enhanced the <it>in vitro </it>light chain activity of BoNT/B, suggesting that BoNT/B may undergo conformational change upon binding some mAbs.</p> <p>Conclusions</p> <p>In addition to determining <it>in vitro </it>inhibition abilities of a panel of mAbs against BoNT/B1-/B5, this work has determined B12.2 and 2B18.2 to be the best mAbs for sample preparation before Endopep-MS. These mAb characterizations also have the potential to assist with mechanistic studies of BoNT/B protection and treatment, which is important for studying alternative therapeutics for botulism.</p

The Artificial Sweetener Splenda Promotes Gut Proteobacteria, Dysbiosis, and Myeloperoxidase Reactivity in Crohn’s Disease–Like Ileitis

We thank John D. Ward and Lindsey N. Kaydo for their technical support and Dr. Wei Xin for the histological scoring of ileitis severity. ARP is an Assistant Professor of Medicine at CWRU School of Medicine. Metagenomic sequencing was conducted in the laboratory of Dr. Skip Virgin at Washington University, School of Medicine, St. Louis, MO. Raw sequencing data files will be available upon request.Peer reviewedPostprin