KCNQ/M currents in sensory neurons: Significance for pain therapy

Neuronal hyperexcitability is a feature of epilepsy and both inflammatory and neuropathic pain. M currents [I-K(M)] play a key role in regulating neuronal excitability, and mutations in neuronal KCNQ2/3 subunits, the molecular correlates of I-K(M), have previously been linked to benign familial neonatal epilepsy. Here, we demonstrate that KCNQ/M channels are also present in nociceptive sensory systems. I-K(M) was identified, on the basis of biophysical and pharmacological properties, in cultured neurons isolated from dorsal root ganglia (DRGs) from 17-d-old rats. Currents were inhibited by the M-channel blockers linopirdine (IC50, 2.1 muM) and XE991 (IC50, 0.26 muM) and enhanced by retigabine (10 muM). The expression of neuronal KCNQ subunits in DRG neurons was confirmed using reverse transcription-PCR and single-cell PCR analysis and by immunofluorescence. Retigabine, applied to the dorsal spinal cord, inhibited C and Adelta fiber-mediated responses of dorsal horn neurons evoked by natural or electrical afferent stimulation and the progressive "windup" discharge with repetitive stimulation in normal rats and in rats subjected to spinal nerve ligation. Retigabine also inhibited responses to intrapaw application of carrageenan in a rat model of chronic pain; this was reversed by XE991. It is suggested that I-K(M) plays a key role in controlling the excitability of nociceptors and may represent a novel analgesic target

Ontogeny of 2-[ 125I]iodomelatonin binding sites in the chicken (Gallus domesticus) kidney and spleen

To understand the possible role of melatonin receptors in the development of renal and immune functions, age-related variations of 2-[ 125I]iodomelatonin binding sites in the chicken kidney and spleen were investigated by radioreceptor assay. Chickens at embryonic day 20, as well as 2 days, 9 days, 2 weeks, 6 weeks, 12 weeks, and 16 weeks after hatching, were kept under a 12 h light : 12 h dark photoperiod and killed at the middle of the light period. Binding sites for 2-[ 125I]iodomelatonin in membrane preparations of the chicken kidney and spleen were present on embryonic day 20. The maximum binding densities (B(max)) in the kidney increased to a peak between 9 days and 2 weeks of age, then progressively decreased. B(max) values of 2-[ 125I]iodomelatonin binding sites in the chicken spleen were lower than in the kidney. The peak density in the chicken spleen was recorded at day 2 after hatching and decreased significantly after 6 weeks of age. There were no significant differences in binding affinities (K(d)) in kidney and spleen of chicken in the different age groups studied. The unity of Hill coefficients of 2-[ 125I]iodomelatonin binding sites of the chicken kidney and spleen in all age groups tested suggested that only a single class of binding sites was present in these tissues during development. It is proposed that the developmental changes in 2-[ 125I]iodomelatonin binding sites in the chicken kidney and spleen may be pertinent to the development of diurnal rhythms of kidney functions and the post-pubertal decline in immune functions of the chicken.published_or_final_versio

New method for the orthogonal labeling and purification of Toxoplasma gondii proteins while inside the host cell

oxoplasma gondii is an obligate intracellular protozoan parasite that is capable of causing severe disease in immunocompromised humans. How T. gondii is able to modulate the host cell to support itself is still poorly understood. Knowledge pertaining to the host-parasite interaction could be bolstered by developing a system to specifically label parasite proteins while the parasite grows inside the host cell. For this purpose, we have created a strain of T. gondii that expresses a mutant Escherichia coli methionyl-tRNA synthetase (MetRSNLL) that allows methionine tRNA to be loaded with the azide-containing methionine analog azidonorleucine (Anl). Anl-containing proteins are susceptible to a copper-catalyzed “click” reaction to attach affinity tags for purification or fluorescent tags for visualization. The MetRSNLL-Anl system labels nascent T. gondii proteins in an orthogonal fashion, labeling proteins only in MetRSNLL-expressing parasites. This system should be useful for nonradioactive pulse-chase studies and purification of nascently translated proteins. Although this approach allows labeling of a diverse array of parasite proteins, secreted parasite proteins appear to be only minimally labeled in MetRSNLL-expressing T. gondii. The minimal labeling of secreted proteins is likely a consequence of the selective charging of the initiator tRNA (and not the elongator methionine tRNA) by the heterologously expressed bacterial MetRS. IMPORTANCE Studying how T. gondii modifies the host cell to permit its survival is complicated by the complex protein environment of the host cell. The approach presented in this article provides the first method for specific labeling of T. gondii proteins while the parasite grows inside the host cell. We show that this approach is useful for pulse-chase labeling of parasite proteins during in vitro growth. It should also be applicable during in vivo infections and in other apicomplexan parasites, including Plasmodium spp

Connecting low- and high-mass star formation: the intermediate-mass protostar IRAS 05373+2349 VLA 2

Until recently, there have been few studies of the protostellar evolution of intermediate-mass (IM) stars, which may bridge the low- and high-mass regimes. This paper aims to investigate whether the properties of an IM protostar within the IRAS 05373+2349 embedded cluster are similar to that of low- and/or high-mass protostars. We carried out Very Large Array as well as Combined Array for Research in Millimeter Astronomy continuum and 12CO(J=1–0) observations, which uncover seven radio continuum sources (VLA 1–7). The spectral index of VLA 2, associated with the IM protostar is consistent with an ionized stellar wind or jet. The source VLA 3 is coincident with previously observed H2 emission line objects aligned in the north–south direction (P.A. −20 to −12◦), which may be either an ionized jet emanating from VLA 2 or (shock-)ionized cavity walls in the large-scale outflow from VLA 2. The position angle between VLA 2 and 3 is slightly misaligned with the large-scale outflow we map at ∼5-arcsec resolution in 12CO (P.A. ∼30◦), which in the case of a jet suggests precession. The emission from the mm core associated with VLA 2 is also detected; we estimate its mass to be 12–23 M , depending on the contribution from ionized gas. Furthermore, the large-scale outflow has properties intermediate between outflows from low- and high-mass young stars. Therefore, we conclude that the IM protostar within IRAS 05373+2349 is phenomenologically as well as quantitatively intermediate between the low- and high-mass domains

Do mangrove forest restoration or rehabilitation activities return biodiversity to pre-impact levels?

Background Mangrove forest restoration and rehabilitation programs are increasingly undertaken to re-establish ecosystem services in the context of community-based biodiversity conservation. Restoration is returning a habitat to the most natural condition, whereas rehabilitation often focuses on optimising ecosystem services alongside biodiversity. With many different restoration and rehabilitation objectives and techniques existing, it is difficult to assess the general effectiveness of restoration and rehabilitation on biodiversity and ecosystem services. This systematic review protocol presents a methodology that will be used to assess the impacts of mangrove forest restoration and rehabilitation on biodiversity and provisioning ecosystem services in a global context. Methods This review will assess studies that have undertaken biodiversity surveys of restored and rehabilitated mangrove forests by comparing them against suitable mature reference mangrove forests within the same region, or surveys prior to degradation of the forest. This review will investigate how the age and initial tree diversity of a restoration or rehabilitation activities determine the effectiveness of these initiatives. Taxa of commercial value to local communities will be assessed to identify whether rehabilitation for optimal ecosystem service provision is likely to conflict with the full restoration of mangrove forests

Kv7/M-type potassium channels in rat skin keratinocytes.

Skin keratinocytes fulfil important signalling and protective functions. Immunocytochemical experiments revealed the unexpected presence of immunoreactivity for the M-type potassium channel subunit Kv7.2 in the keratinocyte layer of intact rat paw skin and in keratinocytes isolated from the skin of 1-day-old rats and cultured in vitro for 3-10 days. Application of the M-channel enhancer retigabine (3-10 μM) to isolated cultured rat keratinocytes: (a) increased outward membrane currents recorded under voltage clamp, (b) produced ~3 mV hyperpolarization at rest, (c) enhanced ~3-fold the release of ATP induced by the TRPV3 agonist carvacrol (1 mM) and (d) increased the amplitude of the carvacrol-induced intracellular Ca(2+) transient measured with Fura-2. The effect of retigabine on ATP release was prevented by the M-channel blocking agent XE991. We conclude that rat skin keratinocytes possess M-channels that, when activated, can modify their physiological properties, with potential significance for their sensory and other biological functions

Ova-looking feminist theory: a call for consideration within health professions education and research

The role of feminist theory in health professions education is often ‘ova-looked’. Gender is one cause of healthcare inequalities within contemporary medicine. Shockingly, according to the World Health Organisation, no European member state has achieved full gender equity in regard to health outcomes. Further, contemporary curricula have not evolved to reflect the realities of a diverse society that remains riddled with inequity. This paper outlines the history of feminist theory, and applies it to health professions education research and teaching, in order to advocate for its continued relevance within contemporary healthcare

R2 imaging of ferritin iron in thalassaemia patients off and on iron-chelation therapy

Myocardial Tissue Characterization: Fat, Hemorrhage & Edema - Poster presentationAccurate assessment of iron burden is crucial for the management of iron-chelation therapy. MRI provides a means to non-invasively assess tissue iron concentration by exploiting the paramagnetic effects of iron on the relaxation rates of solvent protons. The most widely used method is R2* imaging, which has been shown to be sensitive to myocardial iron overload. Recently, a breath-hold fast spin echo sequence has been proposed for fast and accurate imaging of myocardial and hepatic R2. The purpose of this study was to determine which relaxation rates are sensitive to iron-chelation therapy.published_or_final_versionThe 17th Scientific Meeting & Exhibition of the International Society of Magnetic Resonance in Medicine (ISMRM), Honolulu, HI., 18-24 April 2009. In Proceedings of ISMRM 17th Scientific Meeting & Exhibition, 2009, p. 375

Thoughts that breathe, and words that burn: poetic inquiry within health professions education

Qualitative inquiry is increasingly popular in health professions education, and there has been a move to solidify processes of analysis to demystify the practice and increase rigour. Whilst important, being bound too heavily by methodological processes potentially represses the imaginative creativity of qualitative expression and interpretation—traditional cornerstones of the approach. Rigid adherence to analytic steps risks leaving no time or space for moments of ‘wonder’ or emotional responses which facilitate rich engagement. Poetic inquiry, defined as research which uses poetry ‘as, in, [or] for inquiry’, offers ways to encourage creativity and deep engagement with qualitative data within health professions education. Poetic inquiry attends carefully to participant language, can deepen researcher reflexivity, may increase the emotive impact of research, and promotes an efficiency of qualitative expression through the use of ‘razor sharp’ language. This A Qualitative Space paper introduces the approach by outlining how it may be applied to inquiry within health professions education. Approaches to engaging with poetic inquiry are discussed and illustrated using examples from the field’s scholarship. Finally, recommendations for interested researchers on how to engage with poetic inquiry are made, including suggestions as to how to poetize existing qualitative research practices