T-cell trans-synaptic vesicles are distinct and carry greater effector content than constitutive extracellular vesicles

The immunological synapse is a molecular hub that facilitates the delivery of three activation signals, namely antigen, costimulation/corepression and cytokines, from antigen-presenting cells (APC) to T cells. T cells release a fourth class of signaling entities, trans-synaptic vesicles (tSV), to mediate bidirectional communication. Here we present bead-supported lipid bilayers (BSLB) as versatile synthetic APCs to capture, characterize and advance the understanding of tSV biogenesis. Specifically, the integration of juxtacrine signals, such as CD40 and antigen, results in the adaptive tailoring and release of tSV, which differ in size, yields and immune receptor cargo compared with steadily released extracellular vesicles (EVs). Focusing on CD40L+ tSV as model effectors, we show that PD-L1 trans-presentation together with TSG101, ADAM10 and CD81 are key in determining CD40L vesicular release. Lastly, we find greater RNA-binding protein and microRNA content in tSV compared with EVs, supporting the specialized role of tSV as intercellular messenger

Abrogation of mercuric chloride-induced nephritis in the Brown Norway rat by treatment with antibodies against TNFα

HgCl2 induces an autoimmune disease in the Brown Norway rat characterized by synthesis of autoantibodies (mainly, anti-GBM Abs), severe proteinuria and interstitial nephritis. Also, HgCl2- injected rats develop glomerular cell infiltrates consisting of ED1+ cells (monocyte/macrophage), starting on day 4 and reaching a maximum on day 8. Treatment with anti-TNF-α antiserum had preventative effects as it reduced the urinary protein levels to close to the normal range and also blocked the influx of inflammatory cells in the renal glomeruli and interstitium, but circulating anti-GBM and lineal glomerular IgG deposits were unmodified. In addition, whole isolated glomeruli from HgCl2-induced nephritis secreted TNF-α commencing on day 8, being maximally detected on day 11 and preceding, between 2 to 3 days, the development of proteinuria. The administration of anti-TNF-α antiserum or anti-α4 integrin mAb completely abrogated the synthesis of TNF-α in glomeruli isolated from the respective treated groups of animals, in addition to the proteinuria. Taken together our results confirm that TNF-α plays an important role in the induction and development of HgCl2-induced nephritis and highlights the pathogenic importance of the local release of TNF in those renal diseases in which prominent glomerular macrophage accumulation is a constant feature

Phenotypic, Genetic and Environmental Architecture of the Components of Sleep Quality

The genetic and environmental underpinnings of sleep quality have been widely investigated. However, less is known about the etiology of the different sleep quality components and their associations. Subjective sleep quality has been studied most commonly using the Pittsburgh Sleep Quality Index (PSQI). Therefore, this work aimed to study the structure of sleep quality dimensions in a population-based twin sample by examining the etiology of the associations among the PSQI components themselves and between them. The sample comprised 2129 participants from the Murcia Twin Registry. In order to study the phenotypic, genetic and environmental structure of the PSQI we used three alternative multivariate twin models including all seven sub-scales of the PSQI (subjective sleep quality, latency, duration, efficiency, disturbances, use of sleeping medication and daytime dysfunction): a multivariate model (with seven separate correlated factors), a common pathway model and an independent pathway model. The multivariate correlated factors model showed the best fit to the data. All twin models indicated significant genetic overlap among most of the PSQI components, except daytime dysfunction and use of sleep medication. Bivariate heritability explained between 25 and 50% of the covariance for most associations between dimensions. Furthermore, the common pathway model showed that around one third of the variance (0.32; CI 95% 0.18.0.43) of a latent factor common to all questionnaire dimensions is explained by genetic factors. Genetic influences on a latent factor common to all questionnaire dimensions produced the same heritability estimates as the PSQI global score. However, sleep quality dimensions showed considerable specificity regarding its genetic-environmental structure.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. GRANT SUPPORT: Funding: Ministerio de Ciencia, Innovación y Universidades - Spain (RTI2018-095185-B-I00) co-funded by European Regional Development Fund (FEDER)

Role of Tetraspanins CD9 and CD151 in Primary Melanocyte Motility

Tetraspanins CD9 and CD151 have been implicated in cellular motility and intercellular adhesion in several cellular types. Here, we have studied the subcellular localization and the functional role of these molecules in primary melanocytes. We found that endogenous tetraspanins preferentially clustered in areas of melanocyte homotypic intercellular contacts and at the tips of dendrites. These observations were further confirmed using time-lapse fluorescence confocal microscopy of melanocytes transfected with CD9– and CD151–GFP (green fluorescent protein) constructs, suggesting an involvement of these proteins in cellular contacts and migration. Cell adhesion and migration assays performed using blocking monoclonal antibodies against CD9 and CD151 showed no significant effect on cell–extracellular matrix adhesion, whereas the migration of melanocytes was significantly enhanced. The regulation of the migratory capacity of melanocytes by CD9 and CD151 was further confirmed knocking down the endogenous expression of these tetraspanins with small interference RNA oligonucleotides. Therefore, tetraspanin molecules are localized at motile structures in primary human melanocytes regulating the migratory capacity of these cells

Femto-LASIK after Deep Anterior Lamellar Keratoplasty to Correct Residual Astigmatism: A Long-Term Case Series Study

Purpose: To evaluate the long-term outcomes of femtosecond laser-assisted in situ keratomileusis (Femto-LASIK) to correct residual astigmatism after deep anterior lamellar keratoplasty (DALK). Methods: This retrospective case series study included 10 eyes that underwent Femto-LASIK after a DALK. The refractive error, uncorrected (UDVA) and corrected (CDVA) distance visual acuities, thinnest corneal thickness (TCT), and central corneal thickness (CCT) were registered. The postoperative follow-up ranged between 36 and 60 months. Results: All surgeries were uneventful, with no intra- or postoperative complications. The mean UDVA (Snellen scale) rose from 0.13 ± 0.05 to 0.47 ± 0.15 six months after Femto-LASIK (p < 0.001). All cases experienced a significant improvement in UDVA. None of the eyes lost lines of CDVA, and seven eyes (70%) improved the CDVA compared to preoperative values. The refractive cylinder changed from a preoperative value of −3.88 ± 1.00 D to −0.93 ± 0.39 six months after Femto-LASIK (p < 0.0001). In eight eyes (80%), the UDVA and refractive outcomes remained stable at postoperative follow-up visits. In contrast, one eye experienced a refractive regression over the follow-up. TCT and CCT were stable at the different postoperative follow-up visits. Conclusions: Our findings suggest that Femto-LASIK might safely and effectively corrects residual astigmatism after DALK. Despite these encouraging results, further long-term studies, including a larger number of cases, are required to confirm the safety of the procedure. The refractive stability in eyes with prior RK might be lower than for other DALK indications.Depto. de Optometría y VisiónFac. de Óptica y OptometríaTRUEpu

Heritability of sleep quality in a middle-aged twin sample from Spain

©2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ This document is the Accepted version of a Published Work that appeared in final form in Sleep. To access the final edited and published work see https://doi.org/10.1093/sleep/zsy110Study objectives: Sleep quality is associated with health throughout the life span, which is particularly salient in middle-age and older adulthood. Sleep quality appears to be influenced by both genetic and environmental factors. However, there is still limited information about genetic influences on sleep quality in middle-aged adults, and particularly in those from certain geographical locations. We estimated the magnitude of genetic and environmental influences on sleep quality in a representative sample of middle-aged Spanish twins. Methods: The sample comprised 2150 individuals born between 1939 and 1966, who participate in the Murcia Twin Registry. In order to estimate the heritability of sleep quality variables we performed univariate analyses for the global score on the Pittsburgh sleep quality index and for each of its components. Results: We found moderate but significant heritability (34%) for sleep quality. The genetic variance of the components of the Pittsburgh index ranged from 30% to 45%, except for sleep efficiency for which no genetic influence could be detected. In summary, there was a moderate genetic influence on most dimensions of sleep quality in a sample of adult male and female twins. Shared environment influences were not found. Conclusions: This study adds new information regarding the underlying determinants of sleep quality by providing heritability estimates in a middle-aged population-based representative sample from a geographical location that has not been included in studies of this type previously. This could provide a reference point for future research regarding sleep research in middle-age

Three-year follow-up of posterior chamber phakic intraocular lens with a central port design after deep anterior lamellar keratoplasty

Background: To evaluate clinical outcomes of the Visian implantable collamer lens (ICL) with a central port to correct myopia and astigmatism after deep anterior lamellar keratoplasty (DALK) for keratoconus throughout 3 years of follow-up. Methods: This study included 20 eyes of 20 patients that underwent V4c ICL (13 eyes with a spherical ICL and 7 eyes with a toric ICL) implantation after DALK. Uncorrected (UDVA) and corrected (CDVA) distance visual acuities, refraction, intraocular pressure (IOP), endothelial cell density (ECD), and vault were analyzed. Results: The mean UDVA improved from the preoperative 1.18 ± 0.33 logMAR to 0.25 ± 0.14 logMAR at 6 months after surgery (P < 0.0001) and remained unchanged throughout the whole follow-up (P = 0.4). All eyes gained lines of CDVA compared to preoperative values. At the last follow-up visit, all eyes achieved CDVA of 0.2 logMAR or better and 13 eyes (65%) 0.1 logMAR or better. At 6 months post-surgery, all eyes (100%) had a spherical equivalent within ± 1.50 D, and 19 (95%) within ± 1.00 D. The mean manifest spherical equivalent was stable over the postoperative follow-up (P = 0.25). No significant increase in IOP occurred in any case throughout the 3 years of follow-up. The loss in ECD from the preoperative baseline at the last follow-up visit was 2.27%. Conclusions: The clinical outcomes suggest that the V4c ICL implantation for correction of myopia and regular astigmatism in post-DALK eyes was satisfactory in terms of effectiveness, safety, and stability during 3 years of follow-up.Depto. de Optometría y VisiónFac. de Óptica y OptometríaTRUESTAAR® Surgical (Estados Unidos)pu

Gz mediates the long-lasting desensitization of brain CB1 receptors and is essential for cross-tolerance with morphine

Abstract Background Although the systemic administration of cannabinoids produces antinociception, their chronic use leads to analgesic tolerance as well as cross-tolerance to morphine. These effects are mediated by cannabinoids binding to peripheral, spinal and supraspinal CB1 and CB2 receptors, making it difficult to determine the relevance of each receptor type to these phenomena. However, in the brain, the CB1 receptors (CB1Rs) are expressed at high levels in neurons, whereas the expression of CB2Rs is marginal. Thus, CB1Rs mediate the effects of smoked cannabis and are also implicated in emotional behaviors. We have analyzed the production of supraspinal analgesia and the development of tolerance at CB1Rs by the direct injection of a series of cannabinoids into the brain. The influence of the activation of CB1Rs on supraspinal analgesia evoked by morphine was also evaluated. Results Intracerebroventricular (icv) administration of cannabinoid receptor agonists, WIN55,212-2, ACEA or methanandamide, generated a dose-dependent analgesia. Notably, a single administration of these compounds brought about profound analgesic tolerance that lasted for more than 14 days. This decrease in the effect of cannabinoid receptor agonists was not mediated by depletion of CB1Rs or the loss of regulated G proteins, but, nevertheless, it was accompanied by reduced morphine analgesia. On the other hand, acute morphine administration produced tolerance that lasted only 3 days and did not affect the CB1R. We found that both neural mu-opioid receptors (MORs) and CB1Rs interact with the HINT1-RGSZ module, thereby regulating pertussis toxin-insensitive Gz proteins. In mice with reduced levels of these Gz proteins, the CB1R agonists produced no such desensitization or morphine cross-tolerance. On the other hand, experimental enhancement of Gz signaling enabled an acute icv administration of morphine to produce a long-lasting tolerance at MORs that persisted for more than 2 weeks, and it also impaired the analgesic effects of cannabinoids. Conclusion In the brain, cannabinoids can produce analgesic tolerance that is not associated with the loss of surface CB1Rs or their uncoupling from regulated transduction. Neural specific Gz proteins are essential mediators of the analgesic effects of supraspinal CB1R agonists and morphine. These Gz proteins are also responsible for the long-term analgesic tolerance produced by single doses of these agonists, as well as for the cross-tolerance between CB1Rs and MORs.</p