55 research outputs found

    Polycystin-2 (TRPP2) regulates primary cilium length in LLC-PK1 renal epithelial cells

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    Polycystin-2 (PC2, TRPP2) is a Ca2+ permeable nonselective cation channel whose dysfunction generates autosomal dominant polycystic kidney disease (ADPKD). PC2 is present in different cell locations, including the primary cilium of renal epithelial cells. However, little is known as to whether PC2 contributes to the primary cilium structure. Here, we explored the effect(s) of external Ca2+, PC2 channel blockers, and PKD2 gene silencing on the length of primary cilia in wild-type LLC-PK1 renal epithelial cells. Confluent cell monolayers were fixed and immuno-labeled with an anti-acetylated α-tubulin antibody to identify primary cilia and measure their length. Although primary cilia length measurements did not follow a Normal distribution, the data were normalized by Box-Cox transformation rendering statistical differences under all experimental conditions. Cells exposed to high external Ca2+ (6.2 mM) decreased a 13.5% (p < 0.001) primary cilia length as compared to controls (1.2 mM Ca2+). In contrast, the PC2 inhibitors amiloride (200 ΌM) and LiCl (10 mM), both increased primary ciliary length by 33.2% (p < 0.001), and 17.4% (p < 0.001), respectively. PKD2 gene silencing by siRNA elicited a statistically significant, 10.3% (p < 0.001) increase in primary cilia length compared to their respective scrambled RNA transfected cells. The data indicate that conditions that regulate PC2 function or gene expression modify the length of primary cilia in renal epithelial cells. Blocking of PC2 mitigates the effects of elevated external Ca2+ concentration on primary cilia length. Proper regulation of PC2 function in the primary cilium may be essential in the onset of mechanisms that trigger cyst formation in ADPKD.Fil: Scarinci, MarĂ­a Noelia. Consejo Nacional de Investigaciones CientĂ­ficas y TĂ©cnicas. Centro CientĂ­fico TecnolĂłgico Conicet Noa Sur. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo; ArgentinaFil: PĂ©rez, Paula Luciana. Consejo Nacional de Investigaciones CientĂ­ficas y TĂ©cnicas. Centro CientĂ­fico TecnolĂłgico Conicet Noa Sur. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo; ArgentinaFil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones CientĂ­ficas y TĂ©cnicas. Centro CientĂ­fico TecnolĂłgico Conicet Noa Sur. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo; ArgentinaFil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones CientĂ­ficas y TĂ©cnicas. Centro CientĂ­fico TecnolĂłgico Conicet Noa Sur. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, TecnologĂ­a y Desarrollo; Argentin

    Two-dimensional brain microtubule structures behave as memristive devices

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    Microtubules (MTs) are cytoskeletal structures that play a central role in a variety of cell functions including cell division and cargo transfer. MTs are also nonlinear electrical transmission lines that produce and conduct electrical oscillations elicited by changes in either electric field and/or ionic gradients. The oscillatory behavior of MTs requires a voltage-sensitive gating mechanism to enable the electrodiffusional ionic movement through the MT wall. Here we explored the electrical response of non-oscillating rat brain MT sheets to square voltage steps. To ascertain the nature of the possible gating mechanism, the electrical response of non-oscillating rat brain MT sheets (2D arrays of MTs) to square pulses was analyzed under voltage-clamping conditions. A complex voltage-dependent nonlinear charge movement was observed, which represented the summation of two events. The first contribution was a small, saturating, voltage-dependent capacitance with a maximum charge displacement in the range of 4 fC/ÎŒm2. A second, major contribution was a non-saturating voltage-dependent charge transfer, consistent with the properties of a multistep memristive device. The memristive capabilities of MTs could drive oscillatory behavior, and enable voltage-driven neuromorphic circuits and architectures within neurons.Fil: Cantero, MarĂ­a del RocĂ­o. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; ArgentinaFil: Perez, Paula L.. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; ArgentinaFil: Scarinci, MarĂ­a Noelia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; ArgentinaFil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; Argentin

    Correlation between morphological characteristics of macular edema and visual acuity in young patients with idiopathic intermediate uveitis

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    Background and Objectives: Macular edema (ME) is a common complication of intermediate uveitis (IU). It is often responsible for a decrease in visual acuity (VA). Three distinct patterns of macular edema have been described in intermediate uveitis, namely, cystoid macular edema (CME), diffuse macular edema (DME), and serous retinal detachment (SRD). The current study aims to describe the characteristics of macular edema in young patients with idiopathic intermediate uveitis and to correlate its features with VA using spectral domain optical coherence tomography (SD-OCT). Materials and Methods: A total of 27 eyes from 18 patients with idiopathic IU complicated by ME were included in this retrospective study. All patients underwent SD-OCT; data were gathered at the onset of ME. Best-corrected VA (BCVA) was correlated with the morphological features of ME. Results: BCVA was negatively correlated with Ellipsoid Zone (EZ) disruption (p = 0.00021), cystoid pattern (p = 0.00021), central subfield thickness (CST) (p &lt; 0.001), and serous retinal detachment (0.037). Conclusions: In ME secondary to idiopathic IU, VA negatively correlates with Ellipsoid Zone disruption and increases in CST. Moreover, vision is influenced by the presence of cysts in the inner nuclear and outer nuclear layers and by the neuroepithelium detachment

    Stress-induced Perioperative Depressive Symptoms

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    Sustained stress has been correlated with increases in cortisol levels and decreased levels of brain transmitters including serotonin or dopamine. As a result of better understanding of human pathophysiology and pain physiology, the fields of surgery and anesthesia have seen major advances in the last years.However, a high percentage of patients develop depressive symptoms following major surgery and pathogenic perspective is very complex and require an holistic approach

    Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton

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    Dendritic spines (DS) are tiny protrusions implicated in excitatory postsynaptic responses in the CNS. To achieve their function, DS concentrate a high density of ion channels and dynamic actin networks in a tiny specialized compartment. However, to date there is no direct information on DS ionic conductances. Here, we used several experimental techniques to obtain direct electrical information from DS of the adult mouse hippocampus. First, we optimized a method to isolate DS from the dissected hippocampus. Second, we used the lipid bilayer membrane (BLM) reconstitution and patch clamping techniques and obtained heretofore unavailable electrical phenotypes on ion channels present in the DS membrane. Third, we also patch clamped DS directly in cultured adult mouse hippocampal neurons, to validate the electrical information observed with the isolated preparation. Electron microscopy and immunochemistry of PDS-95 and NMDA receptors and intrinsic actin networks confirmed the enrichment of the isolated DS preparation, showing open and closed DS, and multi-headed DS. The preparation was used to identify single channel activities and “whole-DS” electrical conductance. We identified NMDA and Ca2+-dependent intrinsic electrical activity in isolated DS and in situ DS of cultured adult mouse hippocampal neurons. In situ recordings in the presence of local NMDA, showed that individual DS intrinsic electrical activity often back-propagated to the dendrite from which it sprouted. The DS electrical oscillations were modulated by changes in actin cytoskeleton dynamics by addition of the F-actin disrupter agent, cytochalasin D, and exogenous actin-binding proteins. The data indicate that DS are elaborate excitable electrical devices, whose activity is a functional interplay between ion channels and the underlying actin networks. The data argue in favor of the active contribution of individual DS to the electrical activity of neurons at the level of both the membrane conductance and cytoskeletal signaling.Fil: Priel, Avner. Bar-Ilan University; IsraelFil: Dai, Xiao Qing. University of Alberta; CanadáFil: Chen, Xing-Zhen. University of Alberta; CanadáFil: Scarinci, María Noelia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; ArgentinaFil: Cantero, Maria del Rocio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; ArgentinaFil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnologia y Desarrollo.; Argentin

    Retinal Tectonics after Macular Pucker Surgery: Thickness Changes and En-Face Displacement Recovery.

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    PURPOSE To study visual function, retinal layer thickness changes and tangential displacement after Pars Plana Vitrectomy (PPV) for EpiRetinal Membrane (ERM). METHODS Retrospective series of patients undergoing PPV for ERM, with 6-month follow-up including best-corrected visual acuity (BCVA), Optical Coherence Tomography (OCT), M-charts, ERM grading, and InfraRed fundus photo at time 0 (T0, pre-op) at month 1 (T1), 3 (T3), 6 (T6) post-op (±1 week). Retinal layer thickness and tangential (en-face) retinal displacement between successive times for the entire retinal surface and the central horizontal and vertical meridian were also measured. En-face displacement was calculated as optical flow of consecutive images. RESULTS Average BCVA improved from 0.28±0.08 logMAR at T0 to 0.16±0.25 at T6 (p=0.05), BCVA improvement correlated with BVCA at T0 (p<0.001). Vertical metamorphopsia decreased from 1.33°±0.70° at T0 to 0.82°±0.69° at T6 (p<0.05). Foveal thickness reduced from 453±53”m at T0 to 359±31”m at T6 (p<0.05) and reduction correlated with BCVA improvement (p<0.05). Foveal layers decreased (p<0.05) in all cases. Mean en-face deformation was 155.82±50.17”m and mostly occurred in the first month: T0-T1 displacement was 83.59±30.28”m, T1-T3 was 36.28±14.45”m, while T3-T6 was 39.11±22.79”m (p<0.001) on average. Peri-foveal and parafoveal deformation correlated with OCT foveal thickness reduction at all time intervals (1, 3, 6 months: p<0.01). CONCLUSION ERM peeling affects all retinal layer thickness and results in new force balance across the entire retina and tangential displacement. Both en-face and in-depth changes correlate with visual function

    Abdominal drainage after elective colorectal surgery: propensity score-matched retrospective analysis of an Italian cohort

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    background: In italy, surgeons continue to drain the abdominal cavity in more than 50 per cent of patients after colorectal resection. the aim of this study was to evaluate the impact of abdominal drain placement on early adverse events in patients undergoing elective colorectal surgery. methods: a database was retrospectively analysed through a 1:1 propensity score-matching model including 21 covariates. the primary endpoint was the postoperative duration of stay, and the secondary endpoints were surgical site infections, infectious morbidity rate defined as surgical site infections plus pulmonary infections plus urinary infections, anastomotic leakage, overall morbidity rate, major morbidity rate, reoperation and mortality rates. the results of multiple logistic regression analyses were presented as odds ratios (OR) and 95 per cent c.i. results: a total of 6157 patients were analysed to produce two well-balanced groups of 1802 patients: group (A), no abdominal drain(s) and group (B), abdominal drain(s). group a versus group B showed a significantly lower risk of postoperative duration of stay &gt;6 days (OR 0.60; 95 per cent c.i. 0.51-0.70; P &lt; 0.001). a mean postoperative duration of stay difference of 0.86 days was detected between groups. no difference was recorded between the two groups for all the other endpoints. conclusion: this study confirms that placement of abdominal drain(s) after elective colorectal surgery is associated with a non-clinically significant longer (0.86 days) postoperative duration of stay but has no impact on any other secondary outcomes, confirming that abdominal drains should not be used routinely in colorectal surgery

    Psychological Profile and Distinct Salivary Cortisol Awake Response (CAR) in Two Different Study Populations with Obstructive Sleep Apnea (OSA) and Central Serous Chorioretinopathy (CSC)

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    Obstructive sleep apnea (OSA) and central serous chorioretinopathy (CSC) are in terms of nosography different pathologies, however they share a stress-related physio-pathogenetic component, not yet explored in depth. Therefore, the aim of the present study was to ascertain whether OSA and CSC share a common profile, specifically in cortisol production focusing on the cortisol awake response (CAR), the area under curve (AUCCAR) and the SLOPECAR compared with healthy matched controls. Furthermore, standardized self-administered questionnaires were used to identify mental health status related to depression, anxiety and subjective stress perception levels in the study populations. The results showed hypothalamus-pituitary-adrenal (HPA) axis activity anomalies, represented by a flattening CAR in the OSA group and a statistically significant increase in cortisol production in CSC patients at awakening. This disarrangement of the HPA axis activity associated with elevated distress and mental health scores, and its presence in both patients with OSA and patients with CSC, might represent the shared path explaining the stress-related component in these diseases. Further research is needed to investigate the psycho-neuro-endocrinological aspects of OSA and CSC to determine whether psychoeducation on effective stress coping strategies might be of value in improving the quality of life of OSA and CSC patients
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