Single organ metastatic sites in non-small cell lung cancer: Patient characteristics, treatment patterns and outcomes from a large retrospective Canadian cohort.

BACKGROUND: There is a paucity of information about the characteristics, treatment patterns, and outcomes of non-small cell lung cancer (NSCLC) patients with single organ metastasis (SOM). METHODS: This retrospective cohort study includes all patients with a diagnosis of stage IV NSCLC diagnosed from 2014 to 2016 and treated at Princess Margaret Cancer Centre. We compared baseline characteristics and patterns of metastatic sites between patients with SOM versus multiple (M)OM. Additionally, we identified treatment modalities and outcomes for patients with SOM. Cox multivariable models (MVA) were utilized to evaluate differences in overall survival (OS) between the SOM and MOM cohorts. RESULTS: Of 893 pts analyzed, 457 (51 %) had SOM, while 436 (49 %) had MOM at initial diagnosis. Demographics were comparable between the two groups. Brain was the most common site of metastasis for SOM patients. When compared to the MOM group, the SOM group had lower percentages of liver and adrenal metastases. Amongst SOM patients, 54 % received single modality treatment, and 20 % did not receive any treatment for their SOM. In MVA, patients with liver (HR 2.4), bone (HR 1.8), and pleural (HR 1.7) metastasis as their SOM site had the worst outcomes, with median OS of 6.8 months, 12.1 months, and 13.0 months respectively. Patients with SOM had a significantly improved median OS compared to those with MOM (15.9 months vs. 10.6 months; HR 0.56, 95 % CI 0.47-0.66, p < 0.001). CONCLUSION: In NSCLC patients who presented with SOM, survival correlated with the initial organ involved and was better overall compared to patients with MOM. SOM NSCLC may benefit from specific management strategies and SOM patients could be considered as a specific subgroup for survival analyses in observational and non-randomized interventional studies. In clinical trials, SOM can be considered as a stratification factor in the future.

An Automated Imaging-Based Screen for Genetic Modulators of ER Organisation in Cultured Human Cells.

Hereditary spastic paraplegias (HSPs) are a heterogeneous group of mono-genetic inherited neurological disorders, whose primary manifestation is the disruption of the pyramidal system, observed as a progressive impaired gait and leg spasticity in patients. Despite the large list of genes linked to this group, which exceeds 80 loci, the number of cellular functions which the gene products engage is relatively limited, among which endoplasmic reticulum (ER) morphogenesis appears central. Mutations in genes encoding ER-shaping proteins are the most common cause of HSP, highlighting the importance of correct ER organisation for long motor neuron survival. However, a major bottleneck in the study of ER morphology is the current lack of quantitative methods, with most studies to date reporting, instead, on qualitative changes. Here, we describe and apply a quantitative image-based screen to identify genetic modifiers of ER organisation using a mammalian cell culture system. An analysis reveals significant quantitative changes in tubular ER and dense sheet ER organisation caused by the siRNA-mediated knockdown of HSP-causing genes ATL1 and RTN2. This screen constitutes the first attempt to examine ER distribution in cells in an automated and high-content manner and to detect genes which impact ER organisation.

Osteogenic-angiogenic coupled response of cobalt-containing mesoporous bioactive glasses in vivo.

The incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of interest for bone regeneration purposes. In the present work, we have designed a set of bioactive mesoporous glasses SiO2-CaO-P2O5-CoO (Co-MBGs) with different amounts of cobalt. The physicochemical changes introduced by the Co2+ ion, the in vitro effects of Co-MBGs on preosteoblasts and endothelial cells and their in vivo behaviour using them as bone grafts in a sheep model were studied. The results show that Co2+ ions neither destroy mesoporous ordering nor inhibit in vitro bioactive behaviour, exerting a dual role as network former and modifier for CoO concentrations above 3 % mol. On the other hand, the activity of Co-MBGs on MC3T3-E1 preosteoblasts and HUVEC vascular endothelial cells is dependent on the concentration of CoO present in the glass. For low Co-MBGs concentrations (1mg/ml) cell viability is not affected, while the expression of osteogenic (ALP, RUNX2 and OC) and angiogenic (VEGF) genes is stimulated. For Co-MBGs concentration of 5 mg/ml, cell viability decreases as a function of the CoO content. In vivo studies show that the incorporation of Co2+ ions to the MBGs improves the bone regeneration activity of these materials, despite the deleterious effect that this ion has on bone-forming cells for any of the Co-MBG compositions studied. This contradictory effect is explained by the marked increase in angiogenesis that takes place inside the bone defect, leading to an angiogenesis-osteogenesis coupling that compensates for the partial decrease in osteoblast cells. STATEMENT OF SIGNIFICANCE: The development of new bone grafts implies to address the need for osteogenesis-angiogenesis coupling that allows bone regeneration with viable tissue in the long term. In this sense the incorporation of cobalt ions into the composition of bioactive glasses has emerged as a strategy of great interest in this field. Due to the potential cytotoxic effect of cobalt ions, there is an important controversy regarding the suitability of their incorporation in bone grafts. In this work, we address this controversy after the implantation of cobalt-doped mesoporous bioactive glasses in a sheep model. The incorporation of cobalt ions in bioactive glasses improves the bone regeneration ability of these bone grafts, due to enhancement of the angiogenesis-osteogenesis coupling.

Revisión bibliográfica sobre la Gestión Integral de la Farmacoterapia en pacientes hospitalizados en Cirugía Ortopédica y Traumatología / Literature review on Medication Therapy Management in patients admitted to Orthopedic Surgery and Trauma Departments

Objetivo: Explorar la literatura científica sobre las intervenciones farmacéuticas (IF) y la Gestión Integral de la Farmacoterapia (GIF) brindada por farmacéuticos en pacientes hospitalizados en Cirugía Ortopédica y Traumatología (COT). Métodos: Se realizó una búsqueda de artículos originales y revisiones, publicados en inglés o castellano, hasta el 17 de junio de 2021, cuyo tema principal fueran las IF en pacientes hospitalizados en COT. Bases de datos consultadas: Medline, Cochrane Library y Scielo. Términos empleados: “Orthopedics”, “Traumatology”, “orhopedic surgery”, “orhopedic trauma”, “Medication Therapy Management”, “Pharmacy Service, Hospital”, “Pharmacists”, “Medication Reconciliation”, “Pharmaceutical Care”, “Clinical Pharmacy”. La IF fue considerada GIF cuando implicaba una revisión profunda de la medicación del paciente (Valoración inicial/“Assessment”), evaluando indicación, efectividad, seguridad y cumplimiento/adherencia de cada uno de los medicamentos, y comprobando que todas sus necesidades farmacoterapéuticas estuvieran cubiertas. La implantación de la GIF fue considerada completa cuando además se realizaba un plan de cuidados farmacoterapéutico (“Care Plan”) y evaluación/seguimiento (“Follow up-evaluation”). Resultados: Se seleccionaron 29 artículos, principalmente estudios observacionales descriptivos (51,7%). La mayoría fueron publicados en Estados Unidos (27,6%) y España (20,7%). IF mayoritarias: conciliación de medicación (55,2%), revisión de medicación (44,8%), seguimiento farmacoterapéutico (SFT) (34,5%), manejo del dolor físico postquirúrgico (27,6%) y evaluación/reducción de riesgos (27,6%). La GIF fue analizada en cinco referencias bibliográficas (17,2%); siendo su implantación completa solamente en tres (10,3%). Conclusiones: La presente revisión bibliográfica sintetiza las principales IF en pacientes hospitalizados en COT, destacando: conciliación de medicación, revisión de medicación y SFT... (AU)

Objective: To review the scientific literature on Pharmaceutical Interventions (PIs) and Medication Therapy Management (MTM) by pharmacists in patients admitted to Orthopedic Surgery and Trauma (OST) Departments. Methods: A search was conducted of Medline, Cochrane Library, and Scielo databases for original articles and reviews on PIs in adults (≥18 yrs) hospitalized in OST published in English or Spanish up to June 17 2021. Search terms were: “orthopedics”, “traumatology”, “orthopedic surgery”, “orthopedic trauma”, “medication therapy management”, “pharmacy service, hospital”, “pharmacists”, “medication reconciliation”, “pharmaceutical care”, “clinical pharmacy”. Titles and abstracts of retrieved items were examined. A PI was considered MTM when part of a more in-depth review of the patient’s medication, evaluating the indication, effectiveness, safety, and treatment adherence for each medication and ensuring that all pharmacotherapeutic needs were covered (Assessment). MTM implementation was considered complete when plans for pharmacotherapy care (Care Plan) and follow-up (Follow-up evaluation) were developed. Results: The review included 29 articles, mainly descriptive observational studies (51.7%). The majority were published in the USA (27.6%) or Spain (20.7%). The most frequent PIs were: medication reconciliation (55.2%); medication review (44.8%); pharmacotherapeutic follow-up (34.5%); post-surgical physical pain management (27.6%); health education (27.6%); and risk assessment/reduction (27.6%). MTM was analyzed in five retrieved articles (17.2%) and its complete implementation in only three (10.3%). Conclusions: This literature review compiles the main PIs implemented in patients hospitalized in OST, highlighting medication conciliation, medication review, and pharmacotherapeutic follow-up. MTM implementation remains scarce in these patients. (AU)

Effectiveness of a telerehabilitation intervention using ReCOVery APP of long COVID patients: a randomized, 3-month follow-up clinical trial.

The main objective of this study is to analyze the clinical efficacy of telerehabilitation in the recovery of Long COVID patients through ReCOVery APP for 3 months, administered in the Primary Health Care context. The second objective is to identify significant models associated with an improvement in the study variables. An open-label randomized clinical trial was conducted using two parallel groups of a total of 100 Long COVID patients. The first group follows the treatment as usual methods established by their general practitioner (control group) and the second follows the same methods and also uses ReCOVery APP (intervention group). After the intervention, no significant differences were found in favour of the group intervention. Regarding adherence, 25% of the participants made significant use of the APP. Linear regression model establishes that the time of use of ReCOVery APP predicts an improvement in physical function (b = 0.001; p = 0.005) and community social support (b = 0.004; p = 0.021). In addition, an increase in self-efficacy and health literacy also contribute to improving cognitive function (b = 0.346; p = 0.001) and reducing the number of symptoms (b = 0.226; p = 0.002), respectively. In conclusion, the significant use of ReCOVery APP can contribute to the recovery of Long COVID patients. Trial Registration No.: ISRCTN91104012.

Mental health and drug use in college students: Should we take action?

BACKGROUND: College students are vulnerable to suffering from anxiety and depression. Moreover, mental disorders can contribute to drug consumption or inappropriate use of prescribed drugs. Studies on this topic in Spanish college students are limited. This work analyses anxiety and depression and psychoactive drug intake pattern in the post-COVID era in college students. METHODS: An online survey was conducted among college students from UCM (Spain). The survey collected data including demographic, academic student perception, GAD-7 and PHQ-9 scales, and psychoactive substances consumption. RESULTS: A total of 6798 students were included; 44.1 % (CI95%: 42.9 to 45.3) showed symptoms of severe anxiety and 46.5 % (CI95%: 45.4 to 47.8) symptoms of severe or moderately severe depression. The perception of these symptoms did not change after returning to face-to-face university classes in the post-COVID19 era. Despite the high percentage of cases with clear symptoms of anxiety and depression, most students never had a diagnosis of mental illnesses [anxiety 69.2 % (CI95%: 68.1 to 70.3) and depression 78.1 % (CI95%: 77.1 to 79.1)]. Regarding psychoactive substances, valerian, melatonin, diazepam, and lorazepam were the most consumed. The most worrying issue was the consumption of diazepam, 10.8 % (CI95%: 9.8 to 11.8), and lorazepam, 7.7 % (CI95%: 6.9 to 8.6) without medical prescription. Among illicit drugs, cannabis is the most consumed. LIMITATIONS: The study was based on an online survey. CONCLUSIONS: The high prevalence of anxiety and depression aligned with poor medical diagnosis and high intake of psychoactive drugs should not be underestimated. University policies should be implemented to improve the well-being of students.

The emotional well-being of Long COVID patients in relation to their symptoms, social support and stigmatization in social and health services: a qualitative study.

BACKGROUND: Long COVID patients have experienced a decline in their quality of life due to, in part but not wholly, its negative emotional impact. Some of the most prevalent mental health symptoms presented by long COVID patients are anxiety, depression, and sleep disorders. As such, the need has arisen to analyze the personal experiences of these patients to understand how they are managing their daily lives while dealing with the condition. The objective of this study is to increase understanding about the emotional well-being of people diagnosed with long COVID. METHODS: A qualitative design was created and carried out using 35 patients, with 17 participants being interviewed individually and 18 of them taking part in two focus groups. The participating patients were recruited in November and December 2021 from Primary Health Care (PHC) centers in the city of Zaragoza (Northern Spain) and from the Association of Long COVID Patients in Aragon. The study topics were emotional well-being, social support networks, and experience of discrimination. All an inductive thematic content analyses were performed iteratively using NVivo software. RESULTS: The Long COVID patients identified low levels of self-perceived well-being due to their persistent symptoms, as well as limitations in their daily lives that had been persistent for many months. Suicidal thoughts were also mentioned by several patients. They referred to anguish and anxiety about the future as well as a fear of reinfection or relapse and returning to work. Many of the participants reported that they have sought the help of a mental health professional. Most participants identified discriminatory situations in health care. CONCLUSIONS: It is necessary to continue researching the impact that Long COVID has had on mental health, as well as to provide Primary Health Care professionals with evidence that can guide the emotional treatment of these patients.

Sistema inteligente para la gestión de la demanda en atención primaria.

INTRODUCTION AND OBJECTIVES: The Galician Health Service designed a system to improve demand management in primary care known as "XIDE". In it, all professionals participate in an interdisciplinary manner and within their competence framework, to respond to a reason for consultation in a certain time and manner. This article evaluates the pilot phase of implementation of XIDE in primary care of the Galician Health Service. MATERIALS AND METHODS: Cross-sectional descriptive study carried out in 45 primary care centers selected opportunistically at the discretion of the management of the Galician Health Service. For each center, were included all on-demand appointments requested by the adult population in the administrative units in person or by telephone, between 11/2021-05/2022. The XIDE integrates an intelligent search engine that, through algorithms, guides the administrative staff to make an appointment on demand. It performed a descriptive analysis of all the variables, as well as a bivariate analysis with chi-square to identify the causes of the population's rejection of XIDE. RESULTS: The three most frequent reasons for consultation were: knowing the results of the analysis (11.2%), performing blood tests (11.2%) and prescriptions for drugs (10.9%). Family medicine and nursing professionals are the ones who received the most citations. 22.1% of the appointments required to be scheduled on the same day or immediately. The acceptance of the population to the XIDE system was 85.0%. The reason for consultation, response time, mode of care and the recipient professional had a significant influence (p<0.0001) on the rejection of the appointment. CONCLUSIONS: The XIDE adapts globally well to the appointment systems and the organization of primary care of the Galician Health Service, which could facilitate its extension to all health centers in Galicia. However, it is necessary to delve deeper into the causes of rejection in order to introduce improvements that guarantee its viability in the medium-long term.

Enhancing perceived leadership of nursing students through a student-led dedicated education unit in a community setting: A feasibility study.

BACKGROUND: Leadership competence is important to ensure the provision of safe and high-quality care in hospitals. PURPOSE: The aim was to demonstrate the feasibility of enhancing the perceived leadership competence of nursing students through an innovative health education strategy, consisting of a student-led dedicated education unit (DEU) informed by a service-learning approach in a community setting. DESIGN: Feasibility study in a three-phase process: I: Design; II: Acceptability and implementation; III: Expansion to a different setting. In phase II the DEU was piloted with 62 students using a pretest/post-test with control group design. In phase III it was piloted with 20 students in the expansion setting. METHODS: The DEU was implemented and tested in Barcelona (Spain) between 2014 and 2019. To assess acceptability and implementation, students' perceived leadership competence was measured using the Self-Assessment Leadership Instrument. We also obtained satisfaction ratings using an ad hoc questionnaire. RESULTS: Satisfaction was high among students, as well as among service users who received health education. In phase II, students' post-test self-ratings of their leadership competence were higher in both the intervention and control groups, but the increase was significantly greater among students who participated in the DEU (25.84 % vs. 16.72 %, p = .012). Similar results were obtained in the expansion phase, with students in the intervention group once again showing a significant increase in perceived leadership skills (12.89 %, p = .005). CONCLUSIONS: Student-led DEUs appear to be an effective way of enhancing perceived leadership competence among senior nursing students.

Comparative flow cytometry-based immunophenotyping analysis of peripheral blood leukocytes before and after fixation with paraformaldehyde.

Flow cytometry based immunophenotyping provides prime insight into cellular population composition and characteristics, and is widely used in basic and clinical research. Challenges in processing peripheral blood samples in a timely manner necessitate protocol adaptations and utilization of fixatives. Fixation, however, may introduce artifacts to the flow cytometry readout. We performed a comparative flow cytometry immunophenotyping analysis of 13 immune cell populations in the whole blood using a staining protocol with and without fixation step. Freshly procured human peripheral blood samples were stained with a panel of 33 fluorochrome-conjugated antibodies. Samples were processed using a protocol with or without a paraformaldehyde-based fixation step, and matching sample pairs were analyzed by flow cytometry. Our results show that paraformaldehyde-based fixation, in comparison to matched unfixed samples, did not significantly affect population distribution and frequency for: B cells, Plasmablasts, Dendritic cells, NK cells, Granulocytes, Neutrophils, Eosinophils, or Hematopoietic Stem/Progenitor Cells. However, fixation led to significant marker shifts in the subpopulation distribution in CD4, T regulatory, CD8, Monocytes, and Basophils. These results indicate the importance of pre-experimental assessment of fixation-introduced artifacts in the flow cytometry output when considering the feasibility of fresh processing. This is especially important for samples analyzed using comprehensive exploratory immunoprofiling panels.

[Rotator cuff injuries with nighttime pain and sleep quality before and after treatment]. / Lesiones del manguito rotador con dolor nocturno y calidad de sueño antes y después del tratamiento.

INTRODUCTION: Growing evidence suggests that shoulder injuries involving the rotator cuff cause severe pain and deterioration of quality of life and sleep. OBJECTIVE: To present the results of a systematic review on the association of rotator cuff injuries with nighttime pain and sleep quality before and after treatment. MATERIAL AND METHODS: We searched PubMed limited to humans, with no language, age and time period limit with the following terms: [rotator cuff tear and (nocturnal pain OR sleep)]. We used the PRISMA criteria for systematic reviews. Information was extracted on the frequency of nighttime pain and sleep quality in patients with rotator cuff injuries. RESULTS: Of 123 records found, 10 studies were included for meeting criteria, including 1,516 patients. Nighttime pain affected 91-93% of patients; its average intensity was 5.5 points of the EVA. 100% of the studies reported alterations in sleep quality associated with rotator cuff injury. After repair, a decrease in pain to inferior scores of 2 and improvement in sleep quality were reported. CONCLUSION: Rotator cuff injuries produce nighttime pain and sleep quality disturbances that improve with treatment. Alterations in sleep quality are due not only to pain but to alterations in shoulder functionality.

INTRODUCCIÓN: Evidencia creciente sugiere que las lesiones del hombro que involucran al manguito rotador causan dolor severo y deterioro de la calidad de vida y del sueño. OBJETIVO: Presentar los resultados de una revisión sistemática sobre la asociación de lesiones del manguito rotador con dolor nocturno y calidad de sueño antes y después del tratamiento. MATERIAL Y MÉTODOS: Se realizó una búsqueda en PubMed limitada a humanos, sin límite de idioma, edad y período de tiempo con los siguientes términos: [rotator cuff tear and (nocturnal pain OR sleep)]. Se utilizaron los criterios PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses) para revisiones sistemáticas. Se extrajo información sobre la frecuencia de dolor nocturno y calidad del sueño en pacientes con lesiones del manguito rotador. RESULTADOS: De 123 registros encontrados, 10 estudios fueron incluidos por cumplir criterios, incluyendo 1,516 pacientes. El dolor nocturno afectó de 91-93% de los pacientes; su intensidad media fue de 5.5 puntos de la escala visual análoga (EVA). Cien por ciento de los estudios reportaron alteraciones en la calidad de sueño asociado a lesión del manguito rotador. Tras la reparación, en todos los estudios se reportó disminución de dolor a puntuaciones inferiores a 2 y mejoría de la calidad del sueño. CONCLUSIÓN: Las lesiones del manguito rotador producen dolor nocturno y alteraciones de la calidad del sueño que mejoran con el tratamiento. Las alteraciones de la calidad del sueño se deben no sólo a dolor, sino a alteraciones en la funcionalidad del hombro.

Injectable mesoporous bioactive nanoparticles regenerate bone tissue under osteoporosis conditions.

The osteogenic capability of mesoporous bioactive nanoparticles (MBNPs) in the SiO2CaO system has been assessed in vivo using an osteoporotic rabbit model. MBNPs have been prepared using a double template method, resulting in spherical nanoparticles with a porous core-shell structure that has a high surface area and the ability to incorporate the anti-osteoporotic drug ipriflavone. In vitro expression of the pro-inflammatory genes NF-κB1, IL-6, TNF-α, P38 and NOS2 in RAW-264.7 macrophages, indicates that these nanoparticles do not show adverse inflammatory effects. An injectable system has been prepared by suspending MBNPs in a hyaluronic acid-based hydrogel, which has been injected intraosseously into cavitary bone defects in osteoporotic rabbits. The histological analyses evidenced that MBNPs promote bone regeneration with a moderate inflammatory response. The incorporation of ipriflavone into these nanoparticles resulted in a higher presence of osteoblasts and enhanced angiogenesis at the defect site, but without showing significant differences in terms of new bone formation. STATEMENT OF SIGNIFICANCE: Mesoporous bioactive glass nanoparticles have emerged as one of the most interesting materials in the field of bone regeneration therapies. For the first time, injectable mesoporous bioactive nanoparticles have been tested in vivo using an osteoporotic animal model. Our findings evidence that MBG nanoparticles can be loaded with an antiosteoporotic drug, ipriflavone, and incorporated in hyaluronic acid to make up an injectable hydrogel. The incorporation of MBG nanoparticles promotes bone regeneration even under osteoporotic conditions, whereas the presence of IP enhances angiogenesis as well as the presence of osteoblast cells lining in the newly formed bone. The injectable device presented in this work opens new possibilities for the intraosseous treatment of osteoporotic bone using minimally invasive surgery.

Intermittent voluntary wheel running promotes resilience to the negative consequences of repeated social defeat in mice.

A novel approach to reduce the incidence of substance use disorders is to promote resilience to stress using environmental resources such as physical exercise. In the present study we test the hypothesis that Voluntary Wheel Running (VWR) during adolescence blocks the negative consequences of stress induced by intermittent repeated social defeat (IRSD). Four groups of adolescent male C57BL/6 mice were employed in the experiment; two groups were exposed to VWR (1 h, 3 days/week) from postnatal day (PND) 21 until the first social defeat (PND 47), while the remaining two groups did not have access to activity wheels (controls). On PND 47, 50, 53 and 56 mice, who had performed VWR, were exposed to an episode of social defeat by a resident aggressive mouse (VWR+IRSD group) or allowed to explore an empty cage (VWR+EXPL group). The same procedure was performed with control mice that had not undergone VWR (CONTROL+IRSD and CONTROL+EXPL groups). On PND 57, all the mice performed the Elevated Plus Maze (EPM), Hole-Board, Social Interaction, Tail Suspension and Splash tests. After an interval of 3 weeks, all mice underwent a conditioned place preference (CPP) procedure with 1 mg/kg of cocaine. Exposure to VWR prevented the negative consequences of social stress in the EPM, splash test and CPP, since the VWR+IRSD group did not display anxiety- or depression-like effects or the potentiation of cocaine reward observed in the Control+IRSD group. Our results support the idea that physical exercise promotes resilience to stress and represents an excellent target in drug abuse prevention.

Extensions of empirical likelihood and chi-squared-based tests for ordered alternatives.

Several methods for comparing k populations have been proposed in the literature. These methods assess the same null hypothesis of equal distributions but differ in the alternative hypothesis they consider. We focus on two important alternative hypotheses: monotone and umbrella ordering. Two new families of test statistics are proposed, including two known tests, as well as two new powerful tests under monotone ordering. Furthermore, these families are adapted for testing umbrella ordering. We compare some members of the families with respect to power and Type I errors under different simulation scenarios. Finally, the methods are illustrated in several applications to real data.

Brief Maternal Separation Inoculates Against the Effects of Social Stress on Depression-Like Behavior and Cocaine Reward in Mice.

Exposure to intermittent repeated social defeat (IRSD) increases the vulnerability of mice to the rewarding effects of cocaine in the conditioned place preference (CPP) paradigm. According to the "inoculation of stress" hypothesis, a brief period of maternal separation (MS) can provide protection against the negative effects of IRSD. The aim of the present study was to assess whether exposure to a brief episode of MS prevents the subsequent short-term effects of IRSD on depression- and anxiety-like behaviors and to explore its long-term effects on cocaine CPP in mice. Four groups of male C57BL/6 mice were employed; two groups were separated from their mother [6 h on postnatal day (PND) 9], while the other two groups were not (controls). On PND 47, 50, 53 and 56, mice that had experienced MS were exposed to social defeat in the cage of an aggressive resident mouse (MS + IRSD group) or were allowed to explore an empty cage (MS + EXPL group). The same procedure was performed with control mice that had not experienced MS (CONTROL + IRSD and CONTROL + EXPL groups). On PND57-58, all the mice performed the elevated plus maze and the hole-board, social interaction and splash tests. Three weeks after the last episode of defeat, all the mice underwent the CPP procedure with cocaine (1 mg/kg). Irrespective of whether or not MS had taken place, a reduction in open arms measures, dips, and social interaction was observed in mice that experienced IRSD. A higher latency of grooming and acquisition of cocaine-induced CPP were observed only in mice exposed to IRSD alone (CONTROL + IRSD). These results suggest that exposure to a brief episode of stress early in life increases the subsequent resilience of animals to the effects of social stress on vulnerability to cocaine.

Liver X receptor-agonist treatment rescues degeneration in a Drosophila model of hereditary spastic paraplegia.

Hereditary spastic paraplegias (HSPs) are a group of inherited, progressive neurodegenerative conditions characterised by prominent lower-limb spasticity and weakness, caused by a length-dependent degeneration of the longest corticospinal upper motor neurons. While more than 80 spastic paraplegia genes (SPGs) have been identified, many cases arise from mutations in genes encoding proteins which generate and maintain tubular endoplasmic reticulum (ER) membrane organisation. The ER-shaping proteins are essential for the health and survival of long motor neurons, however the mechanisms by which mutations in these genes cause the axonopathy observed in HSP have not been elucidated. To further develop our understanding of the ER-shaping proteins, this study outlines the generation of novel in vivo and in vitro models, using CRISPR/Cas9-mediated gene editing to knockout the ER-shaping protein ADP-ribosylation factor-like 6 interacting protein 1 (ARL6IP1), mutations in which give rise to the HSP subtype SPG61. Loss of Arl6IP1 in Drosophila results in progressive locomotor deficits, emulating a key aspect of HSP in patients. ARL6IP1 interacts with ER-shaping proteins and is required for regulating the organisation of ER tubules, particularly within long motor neuron axons. Unexpectedly, we identified physical and functional interactions between ARL6IP1 and the phospholipid transporter oxysterol-binding protein-related protein 8 in both human and Drosophila model systems, pointing to a conserved role for ARL6IP1 in lipid homeostasis. Furthermore, loss of Arl6IP1 from Drosophila neurons results in a cell non-autonomous accumulation of lipid droplets in axonal glia. Importantly, treatment with lipid regulating liver X receptor-agonists blocked lipid droplet accumulation, restored axonal ER organisation, and improved locomotor function in Arl6IP1 knockout Drosophila. Our findings indicate that disrupted lipid homeostasis contributes to neurodegeneration in HSP, identifying a potential novel therapeutic avenue for the treatment of this disorder.

Identification of extraocular extension in uveal melanoma by ophthalmological ultrasound.

BACKGROUND AND PURPOSE: Uveal melanoma is the most common primary malignant intraocular tumour in the adult population, with a survival rate of 50% despite advances in treatment and knowledge of this disease. The presence of extraocular extension (EE) worsens the prognosis of these patients, so its proper identification can ensure its management and early intervention. Ophthalmological ultrasound is the technique of choice for the diagnosis and follow-up of these patients, both of the anterior EE using ultrasonic biomicroscopy (UBM), and the posterior EE using A and B ultrasound. The aim of this study is to describe the ultrasound characteristics of the BMU and the A and B ultrasound. MATERIAL AND METHODS: A descriptive and retrospective study is carried out on patients diagnosed with uveal melanoma (UM) and EE from 2003 to 2019. The ultrasound characteristics of the local disease and the follow-up after treatment were recorded completely and at each visit. In the case of anterior EE, photographs of the anterior segment and UBM were taken, while those involving the posterior segment were explored under A and B mode ultrasound. All enucleated eyes were sent for anatomopathological study. RESULTS: Ten patients with an average age of 72.3 years were included. The largest proportion of them were medium-sized tumours, followed by large and small ones. The most frequent morphology of the primary tumour was cupuliform. All the EE presented lower internal reflectivity compared to the primary tumour. No trans-scleral connection bridges were found between the primary tumour and the EE in the ultrasound studies. 50% of patients underwent primary enucleation at the time of diagnosis of intraocular MU due to the presence of the EE, and the remaining 50% presented the EE after initial treatment of the primary tumour with I125 brachytherapy. Sixty percent of the patients presented with posterior EE, and were therefore diagnosed with ultrasound A and B. The most frequent histopathological pattern with 87.5% of patients was the epithelioid pattern. DISCUSSION: Ultrasound scanning in patients with MU is mandatory for diagnosis and follow-up of EE. BMU and A and B ultrasound are the test of choice for anterior and posterior EE, respectively. EE have particular ultrasound characteristics such as low internal reflectivity, regularity of their contour and their location usually adjacent to the base of the primary intraocular tumor.

Identificación de extensión extraocular en el melanoma uveal mediante ecografía oftalmológica / Identification of Extraocular Extension in Uveal Melanoma by Ophthalmological Ultrasound

Antecedentes y objetivoEl melanoma uveal es el tumor primario maligno intraocular más frecuente en la población adulta, con una tasa de supervivencia del 50% a pesar de los avances en el tratamiento y conocimiento de esta enfermedad. La presencia de extensión extraocular (EE) empeora el pronóstico de estos pacientes, por lo que su correcta identificación puede asegurar su manejo e intervención temprana. La ecografía oftalmológica es la técnica de elección para el diagnóstico y seguimiento de estos pacientes, tanto de la EE anterior mediante biomicroscopía ultrasónica (UBM), como de la EE posterior mediante ecografía A y B. El objetivo de este estudio es describir las características ecográficas de la UBM y de la ecografía A y B.Material y métodosSe realiza un estudio descriptivo y retrospectivo de los pacientes diagnosticados de melanoma uveal (UM) y EE desde 2003 hasta 2019. Las características ecográficas de la enfermedad local y el seguimiento luego del tratamiento se registraron de maneracompleta y en cada visita. En caso de EE anteriores se realizaron fotografías de segmento anterior y BMU, por el contrario las que involucran el segmento posterior se exploraron bajo ecografía modo A y B. Todos los ojos enucleados se enviaron para su estudio anatomopatológico.

Background and purposeUveal melanoma is the most common primary malignant intraocular tumour in the adult population, with a survival rate of 50% despite advances in treatment and knowledge of this disease. The presence of extraocular extension (EE) worsens the prognosis of these patients, so its proper identification can ensure its management and early intervention. Ophthalmological ultrasound is the technique of choice for the diagnosis and follow-up of these patients, both of the anterior EE using ultrasonic biomicroscopy (UBM), and the posterior EE using A and B ultrasound. The aim of this study is to describe the ultrasound characteristics of the BMU and the A and B ultrasound.Material and methodsA descriptive and retrospective study is carried out on patients diagnosed with uveal melanoma (UM) and EE from 2003 to 2019. The ultrasound characteristics of the local disease and the follow-up after treatment were recorded completely and at each visit. In the case of anterior EE, photographs of the anterior segment and UBM were taken, while those involving the posterior segment were explored under A and B mode ultrasound. All enucleated eyes were sent for anatomopathological study.

Effects of mesenchymal stem cell culture on radio sterilized human amnion or radio sterilized pig skin in burn wound healing.

Deep second and third degree burns treatment requires fibroblasts, keratinocytes and other skin cells in order to grow new dermis and epidermis. Cells can proliferate, secrete growth factors and extracellular matrix required to repair the damaged tissue. Radiosterilized human amnion and radiosterilized pig skin have been used as natural origin skin dressings for burned patients. Adipose-derived mesenchymal stem cells can differentiate into fibroblasts and keratinocytes and improve wound-healing progress. These cells can stimulate vascular tissue formation, release growth factors, synthetize new extracellular matrix and immunoregulate other cells. In this study, we developed mesenchymal stem cells-cellularized skin substitutes based from radiosterilized human amnion or pig skin. Third-degree burns were induced in mice animal models to evaluate the effect of cellularized skin substitutes on burn wound healing. Mesenchymal phenotype was immunophenotypically confirmed by flow cytometry and cell viability was close to 100%. Skin recovery was evaluated in burned mice after seven and fourteen days post-coverage with cellularized and non-cellularized sustitutes. Histological techniques and immunofluorescence were used to evaluate re-epithelization and type I collagen deposition. We determined that cellularized-human amnion or cellularized-pig skin in combination with mesenchymal stem cells improve extracellular matrix deposition. Both cellularized constructs increase detection of type I collagen in newly formed mouse skin and can be potentially used as skin coverage for further clinical treatment of burned patients.

The integrin beta1 modulator Tirofiban prevents adipogenesis and obesity by the overexpression of integrin-linked kinase: a pre-clinical approach in vitro and in vivo.

BACKGROUND: Obesity is caused by the enlargement of the white adipose tissue (WAT) depots, characterized by the hypertrophic enlargement of malfunctioning adipocytes within WAT which increases the storage of triglycerides (TG) in the lipid droplets (LD). Adipogenesis pathways as well as the expression and activity of some extracellular matrix receptors integrins are upregulated. Integrinß1 (INTB1) is the main isoform involved in WAT remodeling during obesity and insulin resistance-related diseases. We recently described Integrin Linked Kinase (ILK), a scaffold protein recruited by INTB1, as an important mediator of WAT remodeling and insulin resistance. As the few approved drugs to fight obesity have brought long-term cardiovascular side effects and given that the consideration of INTB1 and/or ILK modulation as anti-obesogenic strategies remains unexplored, we aimed to evaluate the anti-obesogenic capacity of the clinically approved anticoagulant Tirofiban (TF), stated in preclinical studies as a cardiovascular protector. METHODS: Fully differentiated adipocytes originating from C3H10T1/2 were exposed to TF and were co-treated with specific INTB1 blockers or with siRNA-based knockdown ILK expression. Lipid-specific dyes were used to determine the TG content in LD. The genetic expression pattern of ILK, pro-inflammatory cytokines (MCP1, IL6), adipogenesis (PPARγ, Leptin), thermogenesis (UCP1), proliferation (PCNA), lipid metabolism (FASN, HSL, ATGL), and metabolite transporters (FABP4, FAT, AQP7) were detected using quantitative PCR. Cytoskeletal actin polymerization was detected by confocal microscopy. Immunoblotting was performed to detect INTB1 phosphorylation at Thr788/9 and ILK activity as phosphorylation levels of protein kinase B (AKT) in Ser473 and glycogen synthase kinase 3ß (GSK3ß) at Ser9. TF was intraperitoneally administered once per day to wildtype and ILK knockdown mice (cKDILK) challenged with a high-fat diet (HFD) or control diet (STD) for 2 weeks. Body and WAT weight gains were compared. The expression of ILK and other markers was determined in the visceral epididymal (epi) and inguinal subcutaneous (sc) WAT. RESULTS: TF reduced TG content and the expression of adipogenesis markers and transporters in adipocytes, while UCP-1 expression was increased and the expression of lipases, cytokines or PCNA was not affected. Mechanistically, TF rapidly increased and faded the intracellular phosphorylation of INTB1 but not AKT or GSK3ß. F-actin levels were rapidly decreased, and INTB1 blockade avoided the TF effect. After 24 h, ILK expression and phosphorylation rates of AKT and GSK3ß were upregulated, while ILK silencing increased TG content. INTB1 blockade and ILK silencing avoided TF effects on the TG content and the transcriptional expression of PPARγ and UCP1. In HFD-challenged mice, the systemic administration of TF for several days reduced the weight gain on WAT depots. TF reduced adipogenesis and pro-inflammatory biomarkers and increased lipolysis markers HSL and FAT in epiWAT from HFD, while increased UCP1 in scWAT. In both WATs, TF upregulated ILK expression and activity, while no changes were observed in other tissues. In HFD-fed cKDILK, the blunted ILK in epiWAT worsened weight gain and avoided the anti-obesogenic effect of in vivo TF administration. CONCLUSIONS: ILK downregulation in WAT can be considered a biomarker of obesity establishment. Via an INTB1-ILK axis, TF restores malfunctioning hypertrophied WAT by changing the expression of adipocyte-related genes, increasing ILK expression and activity, and reducing TG storage. TF prevents obesity, a property to be added to its anticoagulant and cardiovascular protective advantages.