TRPA1 modulation by Sigma-1 receptor prevents oxaliplatin-induced painful peripheral neuropathy.

Chemotherapy-induced peripheral neuropathy is a frequent, disabling side effect of anticancer drugs. Oxaliplatin, a platinum compound used in the treatment of advanced colorectal cancer, often leads to a form of chemotherapy-induced peripheral neuropathy characterized by mechanical and cold hypersensitivity. Current therapies for chemotherapy-induced peripheral neuropathy are ineffective, often leading to the cessation of treatment. Transient receptor potential ankyrin 1 (TRPA1) is a polymodal, non-selective cation-permeable channel expressed in nociceptors, activated by physical stimuli and cellular stress products. TRPA1 has been linked to the establishment of chemotherapy-induced peripheral neuropathy and other painful neuropathic conditions. Sigma-1 receptor is an endoplasmic reticulum chaperone known to modulate the function of many ion channels and receptors. Sigma-1 receptor antagonist, a highly selective antagonist of Sigma-1 receptor, has shown effectiveness in a phase II clinical trial for oxaliplatin chemotherapy-induced peripheral neuropathy. However, the mechanisms involved in the beneficial effects of Sigma-1 receptor antagonist are little understood. We combined biochemical and biophysical (i.e. intermolecular Förster resonance energy transfer) techniques to demonstrate the interaction between Sigma-1 receptor and human TRPA1. Pharmacological antagonism of Sigma-1R impaired the formation of this molecular complex and the trafficking of functional TRPA1 to the plasma membrane. Using patch-clamp electrophysiological recordings we found that antagonists of Sigma-1 receptor, including Sigma-1 receptor antagonist, exert a marked inhibition on plasma membrane expression and function of human TRPA1 channels. In TRPA1-expressing mouse sensory neurons, Sigma-1 receptor antagonists reduced inward currents and the firing of actions potentials in response to TRPA1 agonists. Finally, in a mouse experimental model of oxaliplatin neuropathy, systemic treatment with a Sigma-1 receptor antagonists prevented the development of painful symptoms by a mechanism involving TRPA1. In summary, the modulation of TRPA1 channels by Sigma-1 receptor antagonists suggests a new strategy for the prevention and treatment of chemotherapy-induced peripheral neuropathy and could inform the development of novel therapeutics for neuropathic pain.

Piezo2 Mediates Low-Threshold Mechanically Evoked Pain in the Cornea.

Mammalian Piezo2 channels are essential for transduction of innocuous mechanical forces by proprioceptors and cutaneous touch receptors. In contrast, mechanical responses of somatosensory nociceptor neurons evoking pain, remain intact or are only partially reduced in Piezo2-deficient mice. In the eye cornea, comparatively low mechanical forces are detected by polymodal and pure mechanosensory trigeminal ganglion neurons. Their activation always evokes ocular discomfort or pain and protective reflexes, thus being a unique model to study mechanotransduction mechanisms in this particular class of nociceptive neurons. Cultured male and female mouse mechano- and polymodal nociceptor corneal neurons display rapidly, intermediately and slowly adapting mechanically activated currents. Immunostaining of the somas and peripheral axons of corneal neurons responding only to mechanical force (pure mechano-nociceptor) or also exhibiting TRPV1 (transient receptor potential cation channel subfamily V member 1) immunoreactivity (polymodal nociceptor) revealed that they express Piezo2. In sensory-specific Piezo2-deficient mice, the distribution of corneal neurons displaying the three types of mechanically evoked currents is similar to the wild type; however, the proportions of rapidly adapting neurons, and of intermediately and slowly adapting neurons were significantly reduced. Recordings of mechano- and polymodal-nociceptor nerve terminals in the corneal surface of Piezo2 conditional knock-out mice revealed a reduced number of mechano-sensitive terminals and lower frequency of nerve terminal impulse discharges under mechanical stimulation. Eye blinks evoked by von Frey filaments applied on the cornea were lower in Piezo2-deficient mice compared with wild type. Together, our results provide direct evidence that Piezo2 channels support mechanically activated currents of different kinetics in corneal trigeminal neurons and contributes to transduction of mechanical forces by corneal nociceptors.SIGNIFICANCE STATEMENT The cornea is a richly innervated and highly sensitive tissue. Low-threshold mechanical forces activate corneal receptors evoking discomfort or pain. To examine the contribution of Piezo2, a low-threshold mechanically activated channel, to acute ocular pain, we characterized the mechanosensitivity of corneal sensory neurons. By using Piezo2 conditional knock-out mice, we show that Piezo2 channels, present in the cell body and terminals of corneal neurons, are directly involved in acute corneal mechano-nociception. Inhibition of Piezo2 for systemic pain treatment is hindered because of its essential role for mechano-transduction processes in multiple body organs. Still, topical modulation of Piezo2 in the cornea may be useful to selectively relief unpleasant sensations and pain associated with mechanical irritation accompanying many ocular surface disorders.

The Immunosuppressant Macrolide Tacrolimus Activates Cold-Sensing TRPM8 Channels.

TRPM8 is a polymodal, nonselective cation channel activated by cold temperature and cooling agents that plays a critical role in the detection of environmental cold. We found that TRPM8 is a pharmacological target of tacrolimus (FK506), a macrolide immunosuppressant with several clinical uses, including the treatment of organ rejection following transplants, treatment of atopic dermatitis, and dry eye disease. Tacrolimus is an inhibitor of the phosphatase calcineurin, an action shared with cyclosporine. Tacrolimus activates TRPM8 channels in different species, including humans, and sensitizes their response to cold temperature by inducing a leftward shift in the voltage-dependent activation curve. The effects of tacrolimus on purified TRPM8 in lipid bilayers demonstrates conclusively that it has a direct gating effect. Moreover, the lack of effect of cyclosporine rules out the canonical signaling pathway involving the phosphatase calcineurin. Menthol (TRPM8-Y745H)- and icilin (TRPM8-N799A)-insensitive mutants were also activated by tacrolimus, suggesting a different binding site. In cultured mouse DRG neurons, tacrolimus evokes an increase in intracellular calcium almost exclusively in cold-sensitive neurons, and these responses were drastically blunted in Trpm8 KO mice or after the application of TRPM8 antagonists. Cutaneous and corneal cold thermoreceptor endings are also activated by tacrolimus, and tacrolimus solutions trigger blinking and cold-evoked behaviors. Together, our results identify TRPM8 channels in sensory neurons as molecular targets of the immunosuppressant tacrolimus. The actions of tacrolimus on TRPM8 resemble those of menthol but likely involve interactions with other channel residues.SIGNIFICANCE STATEMENT TRPM8 is a polymodal TRP channel involved in cold temperature sensing, thermoregulation, and cold pain. TRPM8 is also involved in the pathophysiology of dry eye disease, and TRPM8 activation has antiallodynic and antipruritic effects, making it a prime therapeutic target in several cutaneous and neural diseases. We report the direct agonist effect of tacrolimus, a potent natural immunosuppressant with multiple clinical applications, on TRPM8 activity. This interaction represents a novel neuroimmune interface. The identification of a clinically approved drug with agonist activity on TRPM8 channels could be used experimentally to probe the function of TRPM8 in humans. Our findings may explain some of the sensory and anti-inflammatory effects described for this drug in the skin and the eye surface.

G protein-coupled receptor signalling potentiates the osmo-mechanical activation of TRPC5 channels.

TRPC5 is an ion channel permeable to monovalent and divalent cations that is widely expressed in different tissues. Although implicated in the control of neurite extension and in the growth cone morphology of hippocampal neurons, as well as in fear-related behaviour, the mechanisms by which TRPC5 is activated remain poorly understood. TRPC5 is known to be activated downstream of Gq-coupled receptors and by membrane stretch, and since there is evidence that mechanical stress may directly activate Gq-coupled receptors, we examined the relationship between the activation of TRPC5 by the type 1 histamine receptor and osmotic stress. Using calcium imaging and patch clamp recordings, we found that a higher proportion of cells expressing TRPC5 respond to hypoosmotic solution when they co-express H1R. This response is associated with a phospholipase C-dependent increase in the cells internal calcium concentration, which is abolished on depletion of calcium stores. We also found that the hypoosmotic stimulus that provokes mechanical stress drives the translocation of TRPC5 to the plasma membrane by a mechanism dependent on PI3K. This increase in TRPC5 at the plasma membrane augments the proportion of cells that respond to hypoosmotic stimulation. Together, these results suggest that hypoosmotic cell-swelling activates Gq-coupled receptors, which in turn enhance the activation of TRPC5 by regulating this channel membrane trafficking. Gq-coupled receptors and TPRC5 are co-expressed in several tissues such as those of the vascular system and in somatosensory neurons, suggesting that this mechanism of TRPC5 activation may have interesting and important implications in arterial pressure sensing and mechanotransduction.

The ion channel TRPM8 is a direct target of the immunosuppressant rapamycin in primary sensory neurons.

BACKGROUND AND PURPOSE: The mechanistic target of rapamycin (mTOR) signalling pathway is a key regulator of cell growth and metabolism. Its deregulation is implicated in several diseases. The macrolide rapamycin, a specific inhibitor of mTOR, has immunosuppressive, anti-inflammatory and antiproliferative properties. Recently, we identified tacrolimus, another macrolide immunosuppressant, as a novel activator of TRPM8 ion channels, involved in cold temperature sensing, thermoregulation, tearing and cold pain. We hypothesized that rapamycin may also have agonist activity on TRPM8 channels. EXPERIMENTAL APPROACH: Using calcium imaging and electrophysiology in transfected HEK293 cells and wildtype or Trpm8 KO mouse DRG neurons, we characterized rapamycin's effects on TRPM8 channels. We also examined the effects of rapamycin on tearing in mice. KEY RESULTS: Micromolar concentrations of rapamycin activated rat and mouse TRPM8 channels directly and potentiated cold-evoked responses, effects also observed in human TRPM8 channels. In cultured mouse DRG neurons, rapamycin increased intracellular calcium levels almost exclusively in cold-sensitive neurons. Responses were markedly decreased in Trpm8 KO mice or by TRPM8 channel antagonists. Cutaneous cold thermoreceptor endings were also activated by rapamycin. Topical application of rapamycin to the eye surface evokes tearing in mice by a TRPM8-dependent mechanism. CONCLUSION AND IMPLICATIONS: These results identify TRPM8 cationic channels in sensory neurons as novel molecular targets of the immunosuppressant rapamycin. These findings may help explain some of its therapeutic effects after topical application to the skin and the eye surface. Moreover, rapamycin could be used as an experimental tool in the clinic to explore cold thermoreceptors.

Differences in the Quality of Life of Patients Recently Diagnosed with Crohn's Disease and Ulcerative Colitis.

Inflammatory bowel diseases (IBD) are chronic diseases, encompassing Crohn's disease (CD) and ulcerative colitis (UC). An IBD diagnosis has an impact on the quality of life of patients; this impact can be different according to the type of disease. OBJECTIVE: This study aimed to analyze the differences in the impact on quality of life in the early stages after diagnosis in patients with CD and UC. PATIENTS AND METHODS: This was an observational, multi-center, and cross-sectional study, with the participation of 156 patients recently diagnosed with IBD (<6 months) from 4 hospitals from the Health Council of the Valencian Community. The patients were assessed through the use of the Inflammatory Bowel Disease Questionnaire (IBDQ-32), which measures the quality of life when living with IBD. RESULTS: The sample was composed of 80 patients with CD (51.0%) and 76 patients with a UC diagnosis. The mean age was 42.3 ± 16.2. The CD patients were more affected (42.5%) in their general quality of life than the UC patients (17.1%) (p = 0.001). In the dimensions of the IBDQ-32, the patients with CD showed significant differences in the systemic, emotional, and social spheres. The bowel dimension scores were similar in both groups. CONCLUSIONS: The patients who were recently diagnosed with CD were more affected regarding their quality of life as compared to those who were diagnosed with UC. Psychological care must be considered to mitigate the impact of an IBD diagnosis.

Iodine status in pregnant women in northern Spain. Effect of iodised salt and iodine supplements on thyroid function.

INTRODUCTION: Iodine deficiency is linked to thyroid dysfunction, particularly in pregnant women. The objective of this study was to ascertain the iodine levels of women in the second trimester of pregnancy, analysing the influence of iodine ingestion on urinary iodine concentration (UIC) and maternal thyroid function. METHODS: A prospective observational study of pregnant women from Health Area IV of Asturias (northern Spain) recruited before 13 weeks of gestation between May and June 2017. A questionnaire on iodine intake was completed at the first visit, and urine and serum samples were collected at baseline and again during the second trimester. UIC, thyroid stimulating hormone (TSH) and free thyroxine (FT4) obtained in the second trimester of gestation were analysed and related to iodine intake. Thyroid autoimmunity was also analysed in half of the pregnant women at baseline. RESULTS: A total of 241 pregnant women were studied. Of these, 56.7% used iodised salt, 46.7% consumed ≥2 servings of dairy products daily and 88.1% took iodine supplements. Median UIC was 191µg/l (135.3-294µg/l), with 68.1% of the women having UIC ≥150µg/l. Only iodised salt consumption provided protection against iodine deficiency (odds ratio 0.35 [0.20-0.63], p=0.001). In women with no autoimmune thyroid disease (n=88), mean levels of TSH were lower in those that consumed iodised salt than in those that did not (respectively, 2.08±0.89mIU/l vs. 2.56±1.02mIU/l, p=0.025). In women with autoimmune thyroid disease (n=30), mean levels of TSH were higher in those that took iodine supplements than in those that did not (respectively, 2.97±1.25mIU/l vs. 1.16±0.41mIU/l, p=0.002). CONCLUSIONS: The pregnant women studied from Health Area IV in Asturias maintain adequate nutritional iodine status in the second trimester of gestation. In our sample, only the consumption of iodised salt was associated with adequate iodine nutrition, without affecting maternal thyroid function. Most of the women used iodine supplements, which was linked to higher levels of TSH in pregnant women with autoimmune thyroid disease.

Membrane-tethered peptides patterned after the TRP domain (TRPducins) selectively inhibit TRPV1 channel activity.

The transient receptor potential vanilloid 1 (TRPV1) channel is a thermosensory receptor implicated in diverse physiological and pathological processes. The TRP domain, a highly conserved region in the C terminus adjacent to the internal channel gate, is critical for subunit tetramerization and channel gating. Here, we show that cell-penetrating, membrane-anchored peptides patterned after this protein domain are moderate and selective TRPV1 antagonists both in vitro and in vivo, blocking receptor activity in intact rat primary sensory neurons and their peripheral axons with mean decline time of 30 min. The most potent lipopeptide, TRP-p5, blocked all modes of TRPV1 gating with micromolar efficacy (IC(50)<10 µM), without significantly affecting other thermoTRP channels. In contrast, its retrosequence or the corresponding sequences of other TRPV channels did not alter TRPV1 channel activity (IC(50)>100 µM). TRP-p5 did not affect the capsaicin sensitivity of the vanilloid receptor. Our data suggest that TRP-p5 interferes with protein-protein interactions at the level of the TRP domain that are essential for the "conformational" change that leads to gate opening. Therefore, these palmitoylated peptides, which we termed TRPducins, are noncompetitive, voltage-independent, sequence-specific TRPV1 blockers. Our findings indicate that TRPducin-like peptides may embody a novel molecular strategy that can be exploited to generate a selective pharmacological arsenal for the TRP superfamily of ion channels.

Neonatal TSH as a marker of iodine nutrition status. Effect of maternal ioduria and thyroid function on neonatal TSH.

INTRODUCTION: Neonatal thyroid stimulating hormone (nTSH) is a marker of iodine nutrition status in the population. The WHO considers a prevalence of less than 3% of nTSH levels greater than 5 mIU/L in samples obtained within 72h from birth indicative of iodine sufficiency. The aim of this study was to determine the prevalence of nTSH levels greater than 5 mIU/L in an iodine-sufficient population and its association with maternal, neonatal and obstetric factors. MATERIALS AND METHODS: A total of 243 pregnant women were recruited between May and June 2017 in our health area. A questionnaire of iodine intake was administered, in addition to determination of ioduria, thyroid function and autoimmunity in the first trimester of gestation. We analysed nTSH levels in samples collected between 48 and 72h post birth and other obstetric and neonatal factors. RESULTS: The mean nTSH level (standard deviation) was 2.43 (1.68 mIU/L), with 7.8% of neonates having levels greater than 5 mIU/L. The highest nTSH levels corresponded to neonates of mothers with insufficient ioduria (P = 0.021) or TSH levels greater than 2.5 mIU/L, in both the case of negative (P = 0.049) and positive (P = 0.006) thyroid autoimmunity results. Maternal ioduria less than 150 µg/L was a risk factor for nTSH levels greater than 5 mIU/L (3.70 [1.06-14.60]; P = 0.046), while a neonatal weight of 2500 g or greater was a protective factor (0.14 [0.02-1.00]; P = 0.038). CONCLUSIONS: The prevalence of nTSH levels greater than 5 mIU/L in our health area was high based on the WHO recommendations. Maternal iodine deficiency was associated with a higher risk of nTSH levels greater than 5 mIU/L. Given that nTSH is currently measured before 72h post birth, we need new cut-off points to keep on using nTSH as a marker of iodine nutritional status.

Expression of the cold thermoreceptor TRPM8 in rodent brain thermoregulatory circuits.

The cold- and menthol-activated ion channel transient receptor potential channel subfamily M member 8 (TRPM8) is the principal detector of environmental cold in mammalian sensory nerve endings. Although it is mainly expressed in a subpopulation of peripheral sensory neurons, it has also been identified in non-neuronal tissues. Here, we show, by in situ hybridization (ISH) and by the analysis of transgenic reporter expression in two different reporter mouse strains, that TRPM8 is also expressed in the central nervous system. Although it is present at much lower levels than in peripheral sensory neurons, we found cells expressing TRPM8 in restricted areas of the brain, especially in the hypothalamus, septum, thalamic reticular nucleus, certain cortices and other limbic structures, as well as in some specific nuclei in the brainstem. Interestingly, positive fibers were also found traveling through the major limbic tracts, suggesting a role of TRPM8-expressing central neurons in multiple aspects of thermal regulation, including autonomic and behavioral thermoregulation. Additional ISH experiments in rat brain demonstrated a conserved pattern of expression of this ion channel between rodent species. We confirmed the functional activity of this channel in the mouse brain using electrophysiological patch-clamp recordings of septal neurons. These results open a new window in TRPM8 physiology, guiding further efforts to understand potential roles of this molecular sensor within the brain.

Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2.

Somatosensory low threshold mechanoreceptors (LTMRs) sense innocuous mechanical forces, largely through specialized axon termini termed sensory nerve endings, where the mechanotransduction process initiates upon activation of mechanotransducers. In humans, a subset of sensory nerve endings is enlarged, forming bulb-like expansions, termed bulbous nerve endings. There is no in vitro human model to study these neuronal endings. Piezo2 is the main mechanotransducer found in LTMRs. Recent evidence shows that Piezo1, the other mechanotransducer considered absent in dorsal root ganglia (DRG), is expressed at low level in somatosensory neurons. We established a differentiation protocol to generate, from iPSC-derived neuronal precursor cells, human LTMR recapitulating bulbous sensory nerve endings and heterogeneous expression of Piezo1 and Piezo2. The derived neurons express LTMR-specific genes, convert mechanical stimuli into electrical signals and have specialized axon termini that morphologically resemble bulbous nerve endings. Piezo2 is concentrated within these enlarged axon termini. Some derived neurons express low level Piezo1, and a subset co-express both channels. Thus, we generated a unique, iPSCs-derived human model that can be used to investigate the physiology of bulbous sensory nerve endings, and the role of Piezo1 and 2 during mechanosensation.

The Prevalence and Risk Factors for Antenatal Depression Among Pregnant Immigrant and Native Women in Spain.

Introduction: The aim was to examine the risk factors of anternatal depression among immigrant and native pregnant women in Spain. Method: A total of 1,524 pregnant women completed the Patient Health Questionnaire and the Postpartum Depression Predictors Inventory-Revised form. Results: The native group reported a lower prevalence (15.2%) compared with immigrant group (25.8%). For immigrants, primiparity, moving, and perceived lack instrumental support from friends or emotional support from partners and family members were significant risk factors. Discussion: The study identified risk factors that can be used for preventive interventions during pregnancy. Significance: Screening and interventions for depression during pregnancy should take migration status into account to maximize effective health care. Also, health providers should consider how migration status can result in different risk factors that affect depression during pregnancy.

Identification of molecular determinants of channel gating in the transient receptor potential box of vanilloid receptor I.

Transient receptor potential vanilloid receptor subtype I (TRPV1) is an ion channel gated by physical and chemical stimuli that belongs to the TRPV protein family. TRPV receptors contain a highly conserved, 6-mer segment near the channel gate, known as the TRP box, whose function remains unknown. Here, we performed an alanine scanning mutagenesis of the TRP box of TRPV1 (IWKLQR) and found that mutation of this motif affected channel gating by raising the free energy of channel activation. Functional characterization of TRPV1 mutants showed that substitution of I696, W697, and R701 by alanine severely affected voltage- and heat-dependent activation and notably reduced the capsaicin responsiveness and tachyphylaxia, while mutation of K698, L699, and Q700 had minor effects. In addition, mutation of I696 to alanine promoted a strong outward rectification at negative membrane potentials, and slowed the kinetics of channel activation. Taken together, our findings suggest that modification of I696, W697, and R701 to alanine altered channel function by affecting events downstream of the initial stimuli-sensing step and imply that intersubunit interactions within the TRP box play an important role in TRPV1 gating.

Characterization of TRPC Channels in a Heterologous System Using Calcium Imaging and the Patch-Clamp Technique.

The family of transient receptor potential (TRPs) channels contains 28 mammalian members, each a unique cellular sensor that responds to a wide variety of external and internal signals. TRP channels are expressed by most mammalian cells, where they are involved in many different physiological functions. Canonical TRP channels (TRPCs) form a family of nonselective cationic channels, although with greater selectivity for Ca2+. This family is made up of seven members (TRPC1-7), all of which contain a TRP box in the carboxyl terminal and 3-4 ankyrin repeats in the amino terminal. While these channels share some similar properties, they display diverse gating mechanisms and are involved in different signaling pathways (Gees M et al., Compr Physiol, 2012). The activation or inhibition of these channels has been studied using different approaches and techniques. Here, we characterize the activation of the TRPC5 channel expressed in a heterologous system, using calcium imaging and the patch-clamp technique in whole-cell configuration.

Endogenous calcium buffers regulate fast exocytosis in the synaptic terminal of retinal bipolar cells.

Calcium-triggered exocytosis at the synapse is suppressed by addition of calcium chelators, but the effects of endogenous Ca(2+) buffers have not been tested. We find that 80% of Ca(2+) binding sites in the synaptic terminal of retinal bipolar cells were associated with mobile molecules that suppressed activation of Ca(2+)-sensitive K(+) channels with an efficiency equivalent to approximately 1.2 mM BAPTA. Removing these buffers caused a 30-fold increase in the number of vesicles released by Ca(2+) tail currents lasting approximately 0.5 ms and a 2-fold increase in the rapidly releasable pool of vesicles (RRP). The effects of BAPTA and EGTA indicate that vesicles comprising the RRP were docked at variable distances from Ca(2+) channels. We propose that endogenous Ca(2+) buffers regulate the size of the RRP by suppressing the release of vesicles toward the periphery of the active zone.

A role of the transient receptor potential domain of vanilloid receptor I in channel gating.

Transient receptor potential vanilloid receptor subtype 1 (TRPV1) is an ionotropic receptor activated by temperature and chemical stimuli. The C-terminal region that is adjacent to the channel gate, recognized as the TRP domain, is a molecular determinant of receptor assembly. However, the role of this intracellular domain in channel function remains elusive. Here, we show that replacement of the TRP domain of TRPV1 with the cognate region of TRPV channels (TRPV2-TRPV6) did not affect receptor assembly and trafficking to the cell surface, although those receptors containing the TRP domain of the distantly related TRPV5 and TRPV6 did not display ion channel activity. Notably, functional chimeras exhibited an impaired sensitivity to the activating stimuli, consistent with a significant contribution of this protein domain to channel function. At variance with TRPV1, voltage-dependent gating of chimeric channels could not be detected in the absence of capsaicin and/or heat. Biophysical analysis of functional chimeras revealed that the TRP domain appears to act as a molecular determinant of the activation energy of channel gating. Together, these findings uncover a role of the TRP domain in intersubunit interactions near the channel gate that contribute to the coupling of stimulus sensing to channel opening.

Hypoosmotic- and pressure-induced membrane stretch activate TRPC5 channels.

Transient receptor potential (TRP) channels mediate a wide array of sensory functions. We investigated the role of TRPC5, a poorly characterized channel widely expressed in the central and peripheral nervous system, as a potential osmosensory protein. Here we show that hypoosmotic stimulation activates TRPC5 channels resulting in a large calcium influx. The response to osmotically induced membrane stretch is blocked by GsMTx-4, an inhibitor of stretch activated ion channels. Direct hypoosmotic activation of TRPC5 is independent of phospholipase C function. However, the osmotic response is inhibited in a cell line in which PIP(2) levels are reduced by regulated overexpression of a lipid phosphatase. The response was restored by increasing intracellular PIP(2) levels through the patch pipette. The mechano-sensitivity of the channel was probed in the whole-cell configuration by application of steps of positive pressure through the patch pipette. Pressure-induced membrane stretch also activated TRPC5 channels, suggesting its role as a transducer of osmo-mechanical stimuli. We also demonstrated the expression of TRPC5 in sensory neurones which together with the osmo-mechanical characteristics of TRPC5 channels suggest its putative role in mechanosensory transduction events.

Joint nociceptor nerve activity and pain in an animal model of acute gout and its modulation by intra-articular hyaluronan.

The mechanisms whereby deposition of monosodium urate (MSU) crystals in gout activates nociceptors to induce joint pain are incompletely understood. We tried to reproduce the signs of painful gouty arthritis, injecting into the knee joint of rats suspensions containing amorphous or triclinic, needle MSU crystals. The magnitude of MSU-induced inflammation and pain behavior signs were correlated with the changes in firing frequency of spontaneous and movement-evoked nerve impulse activity recorded in single knee joint nociceptor saphenous nerve fibers. Joint swelling, mechanical and cold allodynia, and hyperalgesia appeared 3 hours after joint injection of MSU crystals. In parallel, spontaneous and movement-evoked joint nociceptor impulse activity raised significantly. Solutions containing amorphous or needle-shaped MSU crystals had similar inflammatory and electrophysiological effects. Intra-articular injection of hyaluronan (HA, Synvisc), a high-MW glycosaminoglycan present in the synovial fluid with analgesic effects in osteoarthritis, significantly reduced MSU-induced behavioral signs of pain and decreased the enhanced joint nociceptor activity. Our results support the interpretation that pain and nociceptor activation are not triggered by direct mechanical stimulation of nociceptors by MSU crystals, but are primarily caused by the release of excitatory mediators by inflammatory cells activated by MSU crystals. Intra-articular HA decreased behavioral and electrophysiological signs of pain, possibly through its viscoelastic filtering effect on the mechanical forces acting over sensitized joint sensory endings and probably also by a direct interaction of HA molecules with the transducing channels expressed in joint nociceptor terminals.

Incidence, prevalence and risk factors related to anxiety symptoms during pregnancy.

Symptoms of anxiety are one of the most prevalent emotional responses in women during their reproductive phase and especially during pregnancy. OBJECTIVE: Estimate the incidence and prevalence of anxiety throughout the three trimesters of pregnancy in addition to studying the possible risk factors associated with anxiety symptoms. METHOD: A sample of 385 pregnant women participated in a longitudinal study in which the GAD-7 questionnaire was used. RESULTS: Anxiety prevalence was 19.5% in the first trimester. In the second trimester, it was 16.8%, with an incidence of 0.048%. In the third trimester, it was 17.2%, with an incidence of 0.068%. The following predictive factors of anxiety symptoms were identified: being a smoker, presence of previous illness and changes in social relationships. CONCLUSIONS: High incidence and prevalence of anxiety symptoms occur during pregnancy; consequently, applicable preventive policies should be developed.

TSH neonatal como marcador del estado de nutrición de yodo. Influencia de la yoduria y la función tiroidea maternas sobre la TSH neonatal / Neonatal TSH as a marker of iodine nutrition status. Effect of maternal ioduria and thyroid function on neonatal TSH

Introducción: La TSH neonatal (TSHn) es un marcador de nutrición de yodo en la población. La OMS relaciona una prevalencia<3% de TSHn>5mUI/L, obtenida a partir de las 72h del nacimiento, con un adecuado estado nutricional de yodo. El objetivo de este estudio es conocer la prevalencia de TSHn>5mUI/L en una población yodosuficiente y su relación con factores maternos, neonatales y obstétricos. Materiales y métodos: Se reclutaron 243 gestantes entre mayo-junio de 2017 en nuestra área sanitaria. Se realizó un cuestionario sobre consumo de yodo y determinación de yoduria, función y autoinmunidad tiroideas en el primer trimestre de gestación. Se analizó la TSHn entre 48-72h del nacimiento, así como otros factores obstétricos y neonatales. Resultados: La TSHn media fue 2,43±1,68mUI/L, con un 7,8% de neonatos con TSHn>5mUI/L. La TSHn más elevada pertenecía a los neonatos de madres con yodurias insuficientes (p=0,021) o con TSH>2,5mUI/L, tanto en autoinmunidad tiroidea negativa (p=0,049) como positiva (p=0,006). La yoduria materna<150μg/L fue un factor de riesgo de TSHn>5mUI/L (3,70 [1,06-14,60], p=0,046), mientras que el peso neonatal ≥2500g fue un factor protector (0,14 [0,02-1,00], p=0,038). Conclusiones: La prevalencia de TSHn>5mUI/L en nuestra área sanitaria fue elevada, según las recomendaciones de la OMS. Se asoció el déficit de yodo materno con mayor riesgo de TSHn>5mUI/L. Dado que en la actualidad la determinación de la TSHn se realiza antes de las 72h del nacimiento, precisamos de nuevos puntos de corte para continuar empleando la TSHn como marcador de nutrición de yodo. (AU)

Introduction: Neonatal thyroid stimulating hormone (nTSH) is a marker of iodine nutrition status in the population. The WHO considers a prevalence of less than 3% of nTSH levels greater than 5mIU/L in samples obtained within 72h from birth indicative of iodine sufficiency. The aim of this study was to determine the prevalence of nTSH levels greater than 5mIU/L in an iodine-sufficient population and its association with maternal, neonatal and obstetric factors. Materials and methods: A total of 243 pregnant women were recruited between May and June 2017 in our health area. A questionnaire of iodine intake was administered, in addition to determination of ioduria, thyroid function and autoimmunity in the first trimester of gestation. We analysed nTSH levels in samples collected between 48 and 72h post birth and other obstetric and neonatal factors. Results: The mean nTSH level (standard deviation) was 2.43 (1.68mIU/L), with 7.8% of neonates having levels greater than 5mIU/L. The highest nTSH levels corresponded to neonates of mothers with insufficient ioduria (p=.021) or TSH levels greater than 2.5mIU/L, in both the case of negative (p=0.049) and positive (p=0.006) thyroid autoimmunity results. Maternal ioduria greater than 150μg/L was a risk factor for nTSH levels greater than 5mIU/L (3.70 [1.06–14.60]; p=0.046), while a neonatal weight of 2500g or greater was a protective factor (0.14 [0.02–1.00]; p=0.038). Conclusions: The prevalence of nTSH levels greater than 5mIU/L in our health area was high based on the WHO recommendations. Maternal iodine deficiency was associated with a higher risk of nTSH levels less than 5mIU/L. Given that nTSH is currently measured before 72h post birth, we need new cut-off points to keep on using nTSH as a marker of iodine nutritional status. (AU)