When qualitative data contradict quantitative data: diabetes distress in the Chinese-Canadian community.

AIMS: To use both quantitative and qualitative approaches to characterize the diabetes distress profile of Chinese-Canadians with Type 2 diabetes and to better understand their experience of living with diabetes. METHODS: We recruited 40 Chinese-Canadian adults with Type 2 diabetes who completed a Mandarin- or Cantonese-language diabetes education programme in Richmond, British Columbia. Using a mixed-methods sequential explanatory research design, participants first completed a 15-item Chinese version of the Diabetes Distress Scale, which included three subscales: emotional burden, regimen-related distress, and physician distress. The self-report survey was followed by a semi-structured interview that addressed the following diabetes-related topics: perspectives towards the healthcare team, emotional health, diabetes-related concerns and stressors, diabetes diagnosis experience, and sources of social support and diabetes education. RESULTS: The mean (sd) scores for total distress 1.5 (0.5), emotional burden 1.7 (0.7), regimen-related distress 1.4 (0.5), and physician distress 1.4 (0.9), fell within the 'low distress' range (< 2.0). Qualitative analysis of semi-structured interviews showed that some participants were dissatisfied with diabetes care providers and experienced emotional challenges, particularly early in their diagnosis. Other themes that emerged included eating distress, fear of complications, language barriers, and medication concerns. CONCLUSIONS: Not only did the qualitative findings from semi-structured interviews capture aspects of diabetes distress that were not included in the quantitative survey, it also uncovered inconsistencies between the two datasets. To more accurately understand the diabetes distress experience of any ethnic community, both quantitative and qualitative approaches contribute unique value.

Restoration of vision after transplantation of photoreceptors.

Cell transplantation is a potential strategy for treating blindness caused by the loss of photoreceptors. Although transplanted rod-precursor cells are able to migrate into the adult retina and differentiate to acquire the specialized morphological features of mature photoreceptor cells, the fundamental question remains whether transplantation of photoreceptor cells can actually improve vision. Here we provide evidence of functional rod-mediated vision after photoreceptor transplantation in adult Gnat1−/− mice, which lack rod function and are a model of congenital stationary night blindness. We show that transplanted rod precursors form classic triad synaptic connections with second-order bipolar and horizontal cells in the recipient retina. The newly integrated photoreceptor cells are light-responsive with dim-flash kinetics similar to adult wild-type photoreceptors. By using intrinsic imaging under scotopic conditions we demonstrate that visual signals generated by transplanted rods are projected to higher visual areas, including V1. Moreover, these cells are capable of driving optokinetic head tracking and visually guided behaviour in the Gnat1−/− mouse under scotopic conditions. Together, these results demonstrate the feasibility of photoreceptor transplantation as a therapeutic strategy for restoring vision after retinal degeneration.

Melanopsin signalling in mammalian iris and retina.

Non-mammalian vertebrates have an intrinsically photosensitive iris and thus a local pupillary light reflex (PLR). In contrast, it is thought that the PLR in mammals generally requires neuronal circuitry connecting the eye and the brain. Here we report that an intrinsic component of the PLR is in fact widespread in nocturnal and crepuscular mammals. In mouse, this intrinsic PLR requires the visual pigment melanopsin; it also requires PLCß4, a vertebrate homologue of the Drosophila NorpA phospholipase C which mediates rhabdomeric phototransduction. The Plcb4(-/-) genotype, in addition to removing the intrinsic PLR, also essentially eliminates the intrinsic light response of the M1 subtype of melanopsin-expressing, intrinsically photosensitive retinal ganglion cells (M1-ipRGCs), which are by far the most photosensitive ipRGC subtype and also have the largest response to light. Ablating in mouse the expression of both TRPC6 and TRPC7, members of the TRP channel superfamily, also essentially eliminated the M1-ipRGC light response but the intrinsic PLR was not affected. Thus, melanopsin signalling exists in both iris and retina, involving a PLCß4-mediated pathway that nonetheless diverges in the two locations.

Melanopsin and rod-cone photoreceptive systems account for all major accessory visual functions in mice.

In the mammalian retina, besides the conventional rod-cone system, a melanopsin-associated photoreceptive system exists that conveys photic information for accessory visual functions such as pupillary light reflex and circadian photo-entrainment. On ablation of the melanopsin gene, retinal ganglion cells that normally express melanopsin are no longer intrinsically photosensitive. Furthermore, pupil reflex, light-induced phase delays of the circadian clock and period lengthening of the circadian rhythm in constant light are all partially impaired. Here, we investigated whether additional photoreceptive systems participate in these responses. Using mice lacking rods and cones, we measured the action spectrum for phase-shifting the circadian rhythm of locomotor behaviour. This spectrum matches that for the pupillary light reflex in mice of the same genotype, and that for the intrinsic photosensitivity of the melanopsin-expressing retinal ganglion cells. We have also generated mice lacking melanopsin coupled with disabled rod and cone phototransduction mechanisms. These animals have an intact retina but fail to show any significant pupil reflex, to entrain to light/dark cycles, and to show any masking response to light. Thus, the rod-cone and melanopsin systems together seem to provide all of the photic input for these accessory visual functions.

Light adaptation in cat retinal rods.

It has long been an open question whether individual rod receptors in the mammalian retina show any light adaptation. The prevailing evidence so far has suggested that these cells, unlike those in lower vertebrates, adapt little if at all. The experiments on cat rods reported here, however, indicate that this is not really true. Since the cone system in the cat retina has a fairly high light threshold, the rods also need to adapt so that they do not saturate with light before the cones fully take over vision at higher light intensities. In similar experiments, adaptation was found in rods of other mammalian species, including primates.

Calcium-calmodulin modulation of the olfactory cyclic nucleotide-gated cation channel.

Although several ion channels have been reported to be directly modulated by calcium-calmodulin, they have not been conclusively shown to bind calmodulin, nor are the modulatory mechanisms understood. Study of the olfactory cyclic nucleotide-activated cation channel, which is modulated by calcium-calmodulin, indicates that calcium-calmodulin directly binds to a specific domain on the amino terminus of the channel. This binding reduces the effective affinity of the channel for cyclic nucleotides, apparently by acting on channel gating, which is tightly coupled to ligand binding. The data reveal a control mechanism that resembles those underlying the regulation of enzymes by calmodulin. The results also point to the amino-terminal part of the olfactory channel as an element for gating, which may have general significance in the operation of ion channels with similar overall structures.

Central role of the CNGA4 channel subunit in Ca2+-calmodulin-dependent odor adaptation.

Heteromultimeric cyclic nucleotide-gated (CNG) channels play a central role in the transduction of odorant signals and subsequent adaptation. The contributions of individual subunits to native channel function in olfactory receptor neurons remain unclear. Here, we show that the targeted deletion of the mouse CNGA4 gene, which encodes a modulatory CNG subunit, results in a defect in odorant-dependent adaptation. Channels in excised membrane patches from the CNGA4 null mouse exhibited slower Ca2+-calmodulin-mediated channel desensitization. Thus, the CNGA4 subunit accelerates the Ca2+-mediated negative feedback in olfactory signaling and allows rapid adaptation in this sensory system.

Localization of the inositol 1,4,5-trisphosphate receptor in synaptic terminals in the vertebrate retina.

Inositol 1,4,5-trisphosphate (InsP3) mobilizes internal Ca2+ in cells by binding to a receptor protein, which has recently been purified and molecularly cloned. To clarify those neuronal functions that are regulated by InsP3, we have localized this InsP3 receptor protein immunocytochemically in the retina, a neural tissue of well-defined structure and function. Positive staining in neurons is confined almost exclusively to the synaptic layers. Using dissociated retinal neurons, we have further localized the receptor to presynaptic terminals of photoreceptors and bipolar cells, as well as the synaptic processes of amacrine cells. The specific association of InsP3 receptors with synaptic terminals suggests a role for InsP3 in synaptic modulation, especially with respect to transmitter release.

Melanopsin-dependent photoreception provides earliest light detection in the mammalian retina.

BACKGROUND: The visual system is now known to be composed of image-forming and non-image-forming pathways. Photoreception for the image-forming pathway begins at the rods and cones, whereas that for the non-image-forming pathway also involves intrinsically photosensitive retinal ganglion cells (ipRGCs), which express the photopigment melanopsin. In the mouse retina, the rod and cone photoreceptors become light responsive from postnatal day 10 (P10); however, the development of photosensitivity of the ipRGCs remains largely unexplored. RESULTS: Here, we provide direct physiological evidence that the ipRGCs are light responsive from birth (P0) and that this photosensitivity requires melanopsin expression. Interestingly, the number of ipRGCs at P0 is over five times that in the adult retina, reflecting an initial overproduction of melanopsin-expressing cells during development. Even at P0, the ipRGCs form functional connections with the suprachiasmatic nucleus, as assessed by light-induced Fos expression. CONCLUSIONS: The findings suggest that the non-image-forming pathway is functional long before the mainstream image-forming pathway during development.

An unusual cGMP pathway underlying depolarizing light response of the vertebrate parietal-eye photoreceptor.

All cellular signaling pathways currently known to elevate cGMP involve the activation of a guanylyl cyclase to synthesize cGMP. Here we describe an exception to this rule. In the vertebrate parietal eye, the photoreceptors depolarize to light under dark-adapted conditions, unlike rods and cones but like most invertebrate photoreceptors. We report that the signaling pathway for this response involves a rise in intracellular cGMP resulting from an inhibition of the phosphodiesterase that hydrolyzes cGMP. Furthermore, this phosphodiesterase is driven by an active G protein in darkness. These results indicate an antagonistic control of the phosphodiesterase by two G proteins, analogous to the Gs/Gi control of adenylyl cyclase. Our findings demonstrate an unusual phototransduction mechanism and at the same time indicate that signaling involving cyclic nucleotides is more elaborate than previously known.

Olfactory transduction. Tale of an unusual chloride current.

An unusual chloride current seems to play an important role in safeguarding olfactory transduction against an unstable ionic environment, and in nonlinearly amplifying the olfactory signal.

Normal light response, photoreceptor integrity, and rhodopsin dephosphorylation in mice lacking both protein phosphatases with EF hands (PPEF-1 and PPEF-2).

Rhodopsin dephosphorylation in Drosophila is a calcium-dependent process that appears to be catalyzed by the protein product of the rdgC gene. Two vertebrate rdgC homologs, PPEF-1 and PPEF-2, have been identified. PPEF-1 transcripts are present at low levels in the retina, while PPEF-2 transcripts and PPEF-2 protein are abundant in photoreceptors. To determine if PPEF-2 alone or in combination with PPEF-1 plays a role in rhodopsin dephosphorylation and to determine if retinal degeneration accompanies mutation of PPEF-1 and/or PPEF-2, we have produced mice carrying targeted disruptions in the PPEF-1 and PPEF-2 genes. Loss of either or both PPEFs has little or no effect on rod function, as mice lacking both PPEF-1 and PPEF-2 show little or no changes in the electroretinogram and PPEF-2-/- mice show normal single-cell responses to light in suction pipette recordings. Light-dependent rhodopsin phosphorylation and dephosphorylation are also normal or nearly normal as determined by (i) immunostaining of PPEF-2-/- retinas with the phosphorhodopsin-specific antibody RT-97 and (ii) mass spectrometry of C-terminal rhodopsin peptides from mice lacking both PPEF-1 and PPEF-2. Finally, PPEF-2-/- retinas show normal histology at 1 year of age, and retinas from mice lacking both PPEF-1 and PPEF-2 show normal histology at 3 months of age, the latest time examined. These data indicate that, in contrast to loss of rdgC function in Drosophila, elimination of PPEF function does not cause retinal degeneration in vertebrates.

Emergency femoral hernia repair: 13-year retrospective comparison of the three classical open surgical approaches.

PURPOSE AND METHODS: Femoral hernia repairs have been done classically with three different open approaches, namely the Lockwood's (LW), Lotheissen's (LT) and McEvedy's (ME) approaches. Current literature has yet provided any definite conclusion over the best approach in emergency situations. This study aims to evaluate and compare the operative outcomes of these three approaches in emergency situations by retrospectively analyzing 190 cases (76 ME, 33 LT, 81 LW) in 13 years at a regional surgical center. RESULTS: Significantly less laparotomies were required for McEvedy's approach (ME 2.6% vs LT 33.3% vs LW 43.2%, p < 0.001), despite the need for bowel resection appear to be higher (ME 43.4% vs LT 27.3% vs LW 27.2%, p = 0.072). Overall hernia recurrence (p = 0.657) and surgical complication rates (p = 0.585) were similar between the three approaches. Although not reaching statistical significance, it appeared that in patients undergoing McEvedy's operation, mean length of stay was longer (ME 10.1 days vs LT 7.4 days vs LW 9.2 days, p = 0.407) and required more operation time (ME 97.4 min vs LT 72.0 min vs LW 79.0 min, p = 0.222). CONCLUSIONS: All three approaches were safe and effective in repairing femoral hernias in the emergency setting. McEvedy's approach may be superior to others when entry into the peritoneum is anticipated, although it may potentially be associated with longer operation time and hospital stay.

Regulation of sensitivity in vertebrate rod photoreceptors by calcium.

Over the past decade and a half, there have been great advances in our understanding of how light is transduced into electrical signals by the retinal rod and cone photoreceptors in vertebrates. One essential feature of these sensory neurons is their ability to adapt to background illumination, which allows them to function over a broad range of light intensities. This adaptation appears to arise mostly from negative feedback on phototransduction that is mediated by calcium ions. Recent work has suggested that this feedback is fairly complex, and involves several pathways directed at different components of phototransduction. From direct measurements of these feedback pathways in rods, it is possible to evaluate their relative contributions to the overall sensitivity of the cell. At the same time, these feedback mechanisms, as currently known, appear to be sufficient for explaining the change in sensitivity of rods during adaptation to light.

Calcium and light adaptation in retinal photoreceptors.

In the past several years there has been great progress in the understanding of the phototransduction process in retinal photoreceptors. Recently, this knowledge has expanded and we now understand the mechanism of background light adaptation in these cells and the role that calcium has to play.

A rich complexity emerges in phototransduction.

Over the past two decades there has been an explosive growth in our understanding of phototransduction, leading to the development of a comprehensive scheme for the process. On the basis of this scheme the finer details of the process are being elucidated. Additional protein components and pathways have been identified, successful quantitative models of parts of the process have been developed, and a detailed understanding of the molecular basis of physiological function has begun to emerge. Here we summarize the most recent developments.

Isolation and in vitro differentiation of conditionally immortalized murine olfactory receptor neurons.

Two major challenges exist in our understanding of the olfactory system. One concerns the enormous combinatorial code underlying odorant discrimination by odorant receptors. The other relates to neurogenesis and neuronal development in the olfactory epithelium. To address these issues, continuous cell cultures containing olfactory receptor neurons (ORNs) were obtained from olfactory epithelia of H-2K(b)-tsA58 transgenic mice. ORNs were detected and characterized by immunocytochemistry, RT-PCR, and Western blot for the markers Galpha(olf), adenylyl cyclase III, the olfactory cyclic nucleotide-gated channel subunits, and olfactory marker protein. In culture, epidermal growth factor and nerve growth factor stimulated proliferation, and brain-derived neurotrophic factor and neurotrophin-3 induced cellular maturation. Clonal cell lines were isolated by fluorescence-activated cell sorting with anti-neural cell adhesion molecule antibodies, and of 144 single cells plated, 39 clones were expanded, propagated, and stored in liquid nitrogen. All attempts at recovery of clonal lines from frozen stocks have been successful. The most thoroughly characterized clone, 3NA12, expressed ORN markers and responded to stimulation by single odorants. Each odorant activated approximately 1% of cells in a clonal line, and this suggests that many different odorant receptors may be expressed by these clonal cells. Therefore, these cell lines and the method by which they have been obtained represent a significant advance in the generation of olfactory cell cultures and provide a system to investigate odorant coding and olfactory neurogenesis.

Disruption of a retinal guanylyl cyclase gene leads to cone-specific dystrophy and paradoxical rod behavior.

One of two orphan photoreceptor guanylyl cyclases that are highly conserved from fish to mammals, GC-E (or retGC1) was eliminated by gene disruption. Expression of the second retinal cyclase (GC-F) as well as the numbers and morphology of rods remained unchanged in GC-E null mice. However, rods isolated from such mice, despite having a normal dark current, recovered from a light flash markedly faster. Unexpectedly, the a- and b-waves of electroretinograms (ERG) from dark-adapted null mice were suppressed markedly. Cones, initially present in normal numbers in the retina, disappeared by 5 weeks, based on ERG and histology. Thus, the GC-E-deficient mouse defines a model for cone dystrophy, but it also demonstrates that morphologically normal rods display paradoxical behavior in their responses to light.