A centronuclear myopathy-causing mutation in dynamin-2 disrupts neuronal morphology and excitatory synaptic transmission in a murine model of the disease.

AIMS: Dynamin-2 is a large GTPase, a member of the dynamin superfamily that regulates membrane remodelling and cytoskeleton dynamics. Mutations in the dynamin-2 gene (DNM2) cause autosomal dominant centronuclear myopathy (CNM), a congenital neuromuscular disorder characterised by progressive weakness and atrophy of the skeletal muscles. Cognitive defects have been reported in some DNM2-linked CNM patients suggesting that these mutations can also affect the central nervous system (CNS). Here we studied how a dynamin-2 CNM-causing mutation influences the CNS function. METHODS: Heterozygous mice harbouring the p.R465W mutation in the dynamin-2 gene (HTZ), the most common causing autosomal dominant CNM, were used as disease model. We evaluated dendritic arborisation and spine density in hippocampal cultured neurons, analysed excitatory synaptic transmission by electrophysiological field recordings in hippocampal slices, and evaluated cognitive function by performing behavioural tests. RESULTS: HTZ hippocampal neurons exhibited reduced dendritic arborisation and lower spine density than WT neurons, which was reversed by transfecting an interference RNA against the dynamin-2 mutant allele. Additionally, HTZ mice showed defective hippocampal excitatory synaptic transmission and reduced recognition memory compared to the WT condition. CONCLUSION: Our findings suggest that the dynamin-2 p.R465W mutation perturbs the synaptic and cognitive function in a CNM mouse model and support the idea that this GTPase plays a key role in regulating neuronal morphology and excitatory synaptic transmission in the hippocampus.

Significant heterogeneity in the diagnosis and long-term management of Wilson disease: Results from a large multicenter Spanish study.

There is uncertainty regarding Wilson's disease (WD) management. OBJECTIVES: To assess, in a multicenter Spanish retrospective cohort study, whether the approach to WD is homogeneous among centers. METHODS: Data on WD patients followed at 32 Spanish hospitals were collected. RESULTS: 153 cases, 58% men, 20.6 years at diagnosis, 69.1% hepatic presentation, were followed for 15.5 years. Discordant results in non-invasive laboratory parameters were present in 39.8%. Intrahepatic copper concentration was pathologic in 82.4%. Genetic testing was only done in 56.6% with positive results in 83.9%. A definite WD diagnosis (Leipzig score ≥4) was retrospectively confirmed in 92.5% of cases. Chelating agents were standard initial therapy (75.2%) with frequent modifications (57%), particularly to maintenance zinc. Enzyme normalization was not achieved by one third, most commonly in the setting of poor compliance, lack of genetic mutations and/or presence of cardiometabolic risk factors. Although not statistically significant, there were trends for sex differences in number of diagnosed cases, age at diagnosis and biochemical response. CONCLUSIONS: Significant heterogeneity in diagnosis and management of WD patients emerges from this multicenter study that includes both small and large reference centers. The incorporation of genetic testing will likely improve diagnosis. Sex differences need to be further explored.

Confirmatory Factor Analysis of the Transgender Knowledge, Attitudes, and Beliefs (T-KAB) Scale for the Mexican Population.

Transphobia is a result of the widespread lack of knowledge among the general population, together with prejudice based on irrational fear and hatred, of those who do not fit the dominant, socially established gender categories. Little is known about transgender-related knowledge, attitudes, and beliefs among the Mexican population, due to the lack of reliable, valid Spanish-language instruments. This study presents a Spanish translation of the Transgender Knowledge, Attitudes, and Beliefs (T-KAB) Scale and examines its psychometric properties for the Mexican population. A cohort of 501 adult subjects, 337 (67.3%) women and 161 (32.1%) men, drawn from the general population, answered the T-KAB in an anonymous online survey. A confirmatory factor analysis (CFA) and internal consistency measure were used to determine whether its dimensions were reliable and valid for use in a Mexican Spanish language and cultural context. A CFA including the three original dimensions proposed and excluding one item from the T-KAB Scale showed appropriate goodness of fit indices (χ2(180) = 389.41; χ2/df = 2.16, RMSEA = 0.048, CI [0.042, 0.055]; CFI = 0.971; TLI = 0.966), with Cronbach's alpha values over 0.85. The psychometric properties exhibited by the Spanish version of the T-KAB support its use for the assessment of knowledge, attitudes, and beliefs regarding transgender people in the Mexican cultural context. This instrument offers researchers a brief, reliable, valid, and easy self-report measure to use in further studies in Spanish-speaking populations.

The interplay between α7 nicotinic acetylcholine receptors, pannexin-1 channels and P2X7 receptors elicit exocytosis in chromaffin cells.

Pannexin-1 (Panx1) forms plasma membrane channels that allow the exchange of small molecules between the intracellular and extracellular compartments, and are involved in diverse physiological and pathological responses in the nervous system. However, the signaling mechanisms that induce their opening still remain elusive. Here, we propose a new mechanism for Panx1 channel activation through a functional crosstalk with the highly Ca2+ permeable α7 nicotinic acetylcholine receptor (nAChR). Consistent with this hypothesis, we found that activation of α7 nAChRs induces Panx1-mediated dye uptake and ATP release in the neuroblastoma cell line SH-SY5Y-α7. Using membrane permeant Ca2+ chelators, total internal reflection fluorescence microscopy in SH-SY5Y-α7 cells expressing a membrane-tethered GCAMP3, and Src kinase inhibitors, we further demonstrated that Panx1 channel opening depends on Ca2+ signals localized in submembrane areas, as well as on Src kinases. In turn, Panx1 channels amplify cytosolic Ca2+ signals induced by the activation of α7 nAChRs, by a mechanism that seems to involve ATP release and P2X7 receptor activation, as hydrolysis of extracellular ATP with apyrase or blockage of P2X7 receptors with oxidized ATP significantly reduces the α7 nAChR-Ca2+ signal. The physiological relevance of this crosstalk was also demonstrated in neuroendocrine chromaffin cells, wherein Panx1 channels and P2X7 receptors contribute to the exocytotic release of catecholamines triggered by α7 nAChRs, as measured by amperometry. Together these findings point to a functional coupling between α7 nAChRs, Panx1 channels and P2X7 receptors with physiological relevance in neurosecretion.

Virtual Navigation Endoscopically Integrated Into Orbital Trauma.

ABSTRACT: Orbital reconstruction has traditionally been accomplished utilizing different techniques developed for this purpose and has been modified and improved throughout the years. However, there is a high rate of complications associated with eye globe positioning and/or migration of implants, which has created the need to continue to improve these techniques to decrease the frequency of complications. On the other hand, techniques that involve an endoscopic approach and technologies that use virtual surgical planning and three-dimensional model impressions are being used more often to decrease complication rates and seek better surgical outcomes. Combining these 2 methods results in endoscopically integrated virtual surgical navigation. When used for orbital reconstruction, it can be a great alternative and can be useful to decrease the risk of complications associated with this procedure. Therefore, this technical note aims to describe the integration of these 2 techniques into the same instrument to demonstrate the synergy of their benefits when used together.

Restraint of Human Skin Fibroblast Motility, Migration, and Cell Surface Actin Dynamics, by Pannexin 1 and P2X7 Receptor Signaling.

Wound healing is a dynamic process required to maintain skin integrity and which relies on the precise migration of different cell types. A key molecule that regulates this process is ATP. However, the mechanisms involved in extracellular ATP management are poorly understood, particularly in the human dermis. Here, we explore the role, in human fibroblast migration during wound healing, of Pannexin 1 channels and their relationship with purinergic signals and in vivo cell surface filamentous actin dynamics. Using siRNA against Panx isoforms and different Panx1 channel inhibitors, we demonstrate in cultured human dermal fibroblasts that the absence or inhibition of Panx1 channels accelerates cell migration, increases single-cell motility, and promotes actin redistribution. These changes occur through a mechanism that involves the release of ATP to the extracellular space through a Panx1-dependent mechanism and the activation of the purinergic receptor P2X7. Together, these findings point to a pivotal role of Panx1 channels in skin fibroblast migration and suggest that these channels could be a useful pharmacological target to promote damaged skin healing.

[Assessment of massive open online courses for teaching nutrition and preventing obesity]. / Evaluación de cursos en línea, masivos y abiertos de educación en salud y nutrición para prevenir la obesidad.

BACKGROUND: A massive open online course (MOOC) is an online course aimed at unlimited participation and open access via the Web. AIM: To assess the MOOC experience for teaching nutrition, healthy food consumption, physical activity and health promotion to prevent obesity. MATERIAL AND METHODS: Two MOOC courses, (one with two versions), are described with the registration and evaluation instruments such as surveys carried out at the beginning and at the end of the courses. Effective participants in the three versions were 17.456, 11.121 in MOOC1, 2.351 in MOOC1 second version and 3.984 in MOOC2. Their median age was 31 years, 82% were women, 60% were professionals and 12% were foreigners. RESULTS: In the final evaluation of the three courses, 85% to 99% qualified as "very good" or "good" all the surveyed topics. Thirty five percent of participants reported having lack of time, 11% reported problems with internet connectivity and 3.9%, personal or work problems. CONCLUSIONS: This is the first experience with MOOC in health and nutrition to prevent obesity in Chile. Considering the good results and positive evaluation of these courses, we estimate that they are an important tool to prevent obesity and chronic diseases in Chile, Latin America and other regions of the world.

In vitro immunomodulatory activities of peptides derived from Salmo salar NK-lysin and cathelicidin in fish cells.

Antimicrobial peptides (AMPs) are amphipathic peptides, which play an important role in innate defence. These peptides are gene-encoded and either constitutively expressed and/or upregulated during an infection. NK-lysins are AMPs with a three-dimensional globular structure. They are larger molecules, which comprise 74-78 amino acid residues and six conserved cysteine residues forming three disulphide bonds. Cathelicidins are a family of antimicrobial peptides that act as important components of the innate immune system with a broad spectrum of antimicrobial activity and immunomodulatory properties. Although they are widely studied in mammals, little is known about their immunomodulatory function. In the present study, we identified and characterized for the first time four NK-lysin-like transcripts from Atlantic salmon (Salmo salar) based on EST reported sequences. In vitro, NK-lysin derived peptides were able to induce the expression of IL-1ß and IL-8 in Salmo salar head kidney leukocytes. We also tested Salmo salar cathelicidin 1 derived peptide in a similar assay, showing its ability to induce the expression of IFN-γ. These results indicate that NK-lysin and cathelicidin 1 derived peptides are able to modulated immune response, suggesting their potential use to enhance immune response in fish.

Cocaine-Levamisole-Induced Vasculitis/Vasculopathy Syndrome.

PURPOSE OF REVIEW: To understand the clinical spectrum of cocaine-levamisole-induced vasculitis. Worldwide recreational drug consumption is high among the adult population from various social strata. The use of cocaine with levamisole, a frequently added antiparasitic diluent, favors the manifestations of vasculitic lesions, especially in the skin. RECENT FINDINGS: New insights into immunological mechanisms involved in the pathogenesis of the disease. There are still many unknown aspects in the pathogenesis of this disease, such as the immune system interaction with p-ANCAs and the release of inflammatory NETs (neutrophil extracellular traps), which are the origin of auto-antigens and tissue damage, manifesting as vasculitic purpura on the skin. The clinical presentation constitutes a challenge for the clinician to be able to distinguish it from small-vessel vasculitides. This paper intends to improve the understanding of this condition, exhibiting the broad clinical spectrum of local and systemic manifestations of cocaine-levamisole-induced vasculitis, to facilitate a timely diagnosis, in order to take corrective measures and avoid sequelae, along with tissue damage and the consequent deformities and permanent scars.

The contribution of distance education to health promotion in Chile.

The objective of this paper is to present the distance education's contribution to developing health promotion in Chile, through evaluation of a postgraduate certificate program for professionals, and a training course for nurse technicians working in primary healthcare, with an 8-month follow-up after program completion. The program methodology was participatory, interactive and reflective, with mentoring support, exercises, group work and discussions as well as content pertinent to the needs of practice. The evaluation was quali-quantitative with an analysis of the student profile, the implementation process, outcomes at the end of the training and impacts on workplace changes. The results showed a high rate of student approval (87 and 76%), good academic performance and a high level of satisfaction with the methodology and knowledge delivered. The participants' final projects were adapted to local work places realities and were implemented by 62.6% of technicians and 43% of professionals, in addition to changes in work practices that favor health promotion. The level of fulfillment of participants' expectations was very high and the most frequent barriers to implementing the final project were lack of time and personnel, along with minimal support from management and low prioritization of health promotion. This study shows the effectiveness of a distance training model for professionals and technicians that can reach the most remote parts of the country, where there is no access to presencial training, with an educational program centered on work activities and current health challenges.

Connexinopathies: a structural and functional glimpse.

Mutations in human connexin (Cx) genes have been related to diseases, which we termed connexinopathies. Such hereditary disorders include nonsyndromic or syndromic deafness (Cx26, Cx30), Charcot Marie Tooth disease (Cx32), occulodentodigital dysplasia and cardiopathies (Cx43), and cataracts (Cx46, Cx50). Despite the clinical phenotypes of connexinopathies have been well documented, their pathogenic molecular determinants remain elusive. The purpose of this work is to identify common/uncommon patterns in channels function among Cx mutations linked to human diseases. To this end, we compiled and discussed the effect of mutations associated to Cx26, Cx32, Cx43, and Cx50 over gap junction channels and hemichannels, highlighting the function of the structural channel domains in which mutations are located and their possible role affecting oligomerization, gating and perm/selectivity processes.

A high proportion of cells carrying trisomy 12 is associated with a worse outcome in patients with chronic lymphocytic leukemia.

The prognosis of chronic lymphocytic leukemia (CLL) patients displaying trisomy 12 (+12) remains unclear. In this study, we analyzed the influence of the proportion of cells with +12, and other clinical and biologic factors, in time to first therapy (TTFT) and overall survival (OS), in 289 patients diagnosed with CLL carrying +12. Median OS was 129 months. One hundred seventy-four patients (60.2%) presented +12 in <60% of cells. TTFT and OS for this subgroup were longer than for the subgroup with +12 in ≥60% of cells, with a median TTFT of 49 months (CI95%, 39-58) vs 30 months (CI95%, 22-38) (P = 0.001); and a median OS of 159 months (CI95%, 119-182), vs 96 months (CI95%, 58-134) (P = 0.015). Other factors associated with a shorter TTFT were: Binet stage, B symptoms, lymphadenopathy, splenomegaly, high lymphocyte count, 11q-, high ß2 microglobulin, and high LDH. In the multivariate analysis, clinical stage, +12 in ≥60% of cells, high lymphocyte count, B symptoms, and 11q- in addition, resulted of significance in predicting shorter TTFT. Significant variables for OS were: Binet stage, lymphadenopathy, splenomegaly, high LDH, high ß2 microglobulin, 11q-, and CD38. In the multivariate analysis, only Binet stage, 11q-, and high ß2microglobulin significantly predicted shorter OS. CLL with +12 entails a heterogeneous group with intermediate prognosis. However, a high proportion of cells carrying +12 separates a subgroup of patients with poor outcome. Copyright © 2015 John Wiley & Sons, Ltd.

Liver and Spleen Stiffness Measurements by Point Shear Wave Elastography via Acoustic Radiation Force Impulse: Intraobserver and Interobserver Variability and Predictors of Variability in a US Population.

OBJECTIVES: Measurements of liver stiffness and spleen stiffness are useful noninvasive ways to assess fibrosis and portal hypertension in patients with chronic liver disease. One method for assessing stiffness is by point shear wave elastography via acoustic radiation force impulse imaging (ARFI). Its advantage is that sites where stiffness is measured are visualized sonographically. However, its reliability has not been well established, and all studies done to date evaluating the use of ARFI in chronic liver disease have been performed outside the United States. We aimed to characterize the intraobserver and interobserver variability of ARFI-measured liver and spleen stiffness. METHODS: Two hepatologists evaluated unselected hepatology outpatients with ARFI. Exclusions were hepatocellular carcinoma, ascites, a surgical shunt or transjugular intrahepatic portosystemic shunt, portal thrombosis, and cholestatic disease. Each operator obtained 20 measurements from the right liver lobe and spleen. Intraclass correlation coefficients (ICC) were calculated. RESULTS: A total of 177 patients were included: median age, 61 years; 85% male; and 43% obese. Intraobserver ICCs were the same for both observers for liver stiffness (0.89; 95% confidence interval [CI], 0.85-0.92) and spleen stiffness (0.72; 95% CI, 0.61-0.80). Interobserver agreement was excellent for liver stiffness (ICC, 0.85; 95% CI, 0.76-0.90) but not as good for spleen stiffness (ICC, 0.73; 95% CI, 0.60-0.83). A body mass index of 30 kg/m2 or greater, waist circumference of greater than 105 cm, and skin-to-capsule distance of 2 cm or greater negatively affected the ICC for liver stiffness; small spleen size negatively affected the ICC for spleen stiffness. CONCLUSIONS: To our knowledge, this article is the first report of ARFI findings in a US population with chronic liver disease. Liver stiffness reproducibility was excellent, particularly in nonobese patients. Spleen stiffness reproducibility was excellent in those with larger spleens and therefore may be most useful in patients with cirrhosis and portal hypertension.

[The debate on regulating biotechnology drugs: Colombia in the international context]. / El debate de la regulación de medicamentos biotecnológicos: Colombia en el contexto internacional.

In September 2014, Colombia issued standards for the evaluation of biological drugs within the framework of the marketing authorization process. The Colombian approach explicitly includes a fast track for evaluating competing biologicals, which caused great national and international controversy. This article explains the context that justifies the need for this fast-track approach, critically analyzes comparability as a paradigm for the evaluation of biogenerics, and shows that Colombia's position is not isolated and is based on global regulatory trends.

[Evaluation of an online health promotion Diploma]. / Evaluación de un programa de educación a distancia en Promoción de Salud para la Atención Primaria en Chile.

BACKGROUND: Health promotion in Chile is essential, considering its current epidemiologic status where chronic diseases are predominant. AIM: To report the evaluation of an on-line Diploma in health promotion for Primary Health Care professionals. MATERIAL AND METHODS: Two hundred sixty four professionals from all regions of the country (40% rural and 60% urban) took the Diploma. It lasted 8 months with 248 academic hours distributed in 5 modules with a total of 15 units. The program was evaluated with four surveys answered by the students (general description of participants, mid-term, final and follow-up). RESULTS: Students were highly satisfied with the program and especially with its interactive methodology which included tutorials, individual and group assignments, online discussions and a final project. The drawbacks were time restrictions and limited internet access. Ninety percent of students completed the course with an overall mean grade of 5.57 (out of a 1-7 scale). The follow-up survey showed that students implemented the new health promotion knowledge acquired, and put in practice their final assignment in their local counties. Also, most students improved their working conditions. CONCLUSIONS: The on-line interactive and participative methodology applied in this Diploma had positive results and the evaluation model can be applied in other health promotion on-line education programs.

Tuberculosis infection causing intestinal perforations in 2 patients with systemic lupus erythematosus.

Patients with systemic lupus erythematosus (SLE) have a higher incidence rate of tuberculosis and a more frequent extrapulmonary involvement than the general population. We present 2 SLE patients who developed gastrointestinal tuberculosis complicated with intestinal perforation, a rare but serious complication that could be confused with lupus-associated intestinal vasculitis. Opportunistic infections such as tuberculosis must be suspected in SLE patients with abdominal symptoms on immunosuppressive therapy because its early recognition could prevent catastrophic complications such as intestinal perforation and subsequent peritonitis.

Biological relapse in multiple myeloma: Outcome and treatment strategies in a Spanish real-world setting.

Recommendations regarding the best time to start treatment in patients with relapsed/refractory multiple myeloma (RRMM) after biological relapse/progression (BR) are unclear. This observational, prospective, multicenter registry aimed to evaluate the impact on time to progression (TTP) of treatment initiation at BR versus at symptomatic clinical relapse (ClinR) based on the Spanish routine practice in adult patients with RRMM. Patients had two or less previous treatment lines and at least one previous partial response. Baseline characteristics and treatment outcomes were recorded, and survival was analyzed. Of 225 patients, 110 were treated at BR (TxBR group) and 115 at ClinR (TxClinR group) according to the investigators' criteria. The proportion of patients with higher ECOG, previous noncomplete remission (CR), and second relapse were significantly higher in the TxBR group compared to the TxClinR group. TheTxClinR group showed improved outcomes, including TTP, compared to the TxBR group. Progression-free survival increased in the TxClinR group (56.2 months) compared to the TxBR group (32.5 months) (p = 0.0137), and median overall survival also increased (p = 0.0897). Median TTP was significantly longer in patients relapsing from a CR (50.4 months) and in their first relapse (38.7 months) compared to those relapsing from a non-CR response (32.9 months) and in their second relapse (25.2 months). Physicians seemed to start treatment earlier in RRMM patients with poor prognosis features. Previous responses to anti-MM treatment and the number of prior treatment lines were identified as prognosis factors, whereby relapse from CR and first relapse were associated with a longer time to progression.

Reduced protein stability of human DJ-1/PARK7 L166P, linked to autosomal recessive Parkinson disease, is due to direct endoproteolytic cleavage by the proteasome.

Parkinson's disease (PD) is characterized by dopaminergic dysfunction and degeneration. DJ-1/PARK7 mutations have been linked with a familial form of early onset PD. In this study, we found that human DJ-1 wild type and the missense mutants M26I, R98Q, A104T and D149A were stable proteins in cells, only the L166P mutant was unstable. In parallel, the former were not degraded and the L166P mutant was directly degraded in vitro by proteasome-mediated endoproteolytic cleavage. Furthermore, genetic evidence in fission yeast showed the direct involvement of proteasome in the degradation of human DJ-1 L166P and the corresponding L169P mutant of SPAC22E12.03c, the human orthologue of DJ-1 in Schizosaccharomyces Pombe, as their protein levels were increased at restrictive temperature in fission yeast (mts4 and pts1-732) harboring temperature sensitive mutations in proteasomal subunits. In total, our results provide evidence that direct proteasomal endoproteolytic cleavage of DJ-1 L166P is the mechanism of degradation contributing to the loss-of-function of the mutant protein, a property not shared by other DJ-1 missense mutants associated with PD.

The association between academic pressure and adolescent mental health problems: A systematic review.

BACKGROUND: Academic pressure is a potential contributor to adolescent mental health problems, but international evidence on this association has never been synthesised. METHODS: We conducted the first systematic review of the association between academic pressure and adolescent depression, anxiety, self-harm, suicidality, suicide attempts and suicide. We searched MEDLINE, PsycINFO, ERIC and Web of Science (core collection) up to November 24, 2022, for studies of school-going children or adolescents, which measured academic pressure or timing within the school year as the exposure and depression, anxiety, self-harm, or suicidal ideation, attempts or suicide as outcomes. Risk of bias was assessed using the Mixed Methods Appraisal Tool. We used narrative synthesis to summarise the evidence. The review was prospectively registered with PROSPERO (CRD42021232702). RESULTS: We included 52 studies. Most studies assessed mixed anxiety and depressive symptoms (n = 20) or depressive symptoms (n = 19). Forty-eight studies found evidence of a positive association between academic pressure or timing within the school year and at least one mental health outcome. LIMITATIONS: Most studies were cross-sectional (n = 39), adjusted for a narrow range of confounders or had other limitations which limited the strength of causal inferences. CONCLUSIONS: We found evidence that academic pressure is a potential candidate for public health interventions which could prevent adolescent mental health problems. Large population-based cohort studies are needed to investigate whether academic pressure is a causal risk factor that should be targeted in school- and policy-based interventions. FUNDING: UCL Health of the Public; Wellcome Institutional Strategic Support Fund.