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Galectins are soluble glycan-binding proteins that interact with a wide range of glycoproteins and glycolipids and modulate a broad spectrum of physiological and pathological processes. The expression and subcellular localization of different galectins vary among tissues and cell types and change during processes of tissue repair, fibrosis and cancer where epithelial cells loss differentiation while acquiring migratory mesenchymal phenotypes. The epithelial-mesenchymal transition (EMT) that occurs in the context of these processes can include modifications of glycosylation patterns of glycolipids and glycoproteins affecting their interactions with galectins. Moreover, overexpression of certain galectins has been involved in the development and different outcomes of EMT. This review focuses on the roles and mechanisms of Galectin-1 (Gal-1), Gal-3, Gal-4, Gal-7 and Gal-8, which have been involved in physiologic and pathogenic EMT contexts., (© 2024. The Author(s).)

(1) Background: Mindfulness-based interventions (MBIs) have shown interesting preliminary effects on self-regulation processes in early adolescence. However, programs have typically combined different types of interventions with no understanding of the specific effect of each intervention type on attentional, emotional, and behavioral regulation. The objective of this research was to evaluate the effect of two MBIs-one focused on classic attentional practices and another focused on the recognition and expression of emotions-on attentional, emotional, and behavioral self-regulation in early adolescents. (2) Method: An experimental paradigm was used. A sample of 74 children aged between 8 and 12 years old were randomly assigned to three experimental conditions: (1) an MBI with a focus on attentional practices, (2) an MBI with a focus on recognition and expression of emotions, and (3) a control group. The interventions lasted 8 weeks, with a weekly, 1 h online synchronous session plus home practices. Children were evaluated before starting the intervention and at the end of the 8-week period. The assessed outcomes were (1) mindfulness; (2) emotional regulation; (3) attentional regulation, and (4) behavioral regulation. (3) Results: Children who participated in both intervention programs increased their mindfulness and emotional and behavioral regulation scores. Only children who participated in the MBI with a focus on attention showed significant changes in their ability to self-regulate attention. (4) Conclusions: The use of online MBIs, with attention to external and internal stimuli practices, can be a good strategy to strengthen self-regulation skills for attention, emotions, and behavior in early adolescence.

Family functioning, understood as cohesion and adaptability, is critical in families with adolescent children, given the changes that this stage implies at the family level. Time perspective is one variable that can facilitate better family functioning through the way people give meaning to the process they live. In this study, we examined the relationship between family functioning and the time perspective of adolescent children's parents. The FACES IV and ZTPI were administered to 276 parents of adolescents. Regression analyses indicated that the past positive, past negative, and future scores predicted family cohesion and adaptability, explaining at least 20% of the variance. Balanced families, with greater cohesion and adaptability, presented a higher level of past positive and future-oriented temporal perspectives, compared to unbalanced families, which presented a greater orientation to the past negative and deviated from the balanced temporal profile. The importance of considering the inter-relationship between family functioning and time perspective was discussed, considering its impact on the health and well-being of families with adolescents.

Intervention programs based on self-compassion have demonstrated their efficacy both in reducing psychological distress and increasing well-being. The goal of this study was to test the efficacy of an online intervention to increase mindfulness and self-compassion levels in a non-clinical sample in a highly stressful context: the ten weeks of lockdown imposed in the early stages of the COVID-19 pandemic. The intervention sessions consisted of thirty-minute guided meditations followed by thirty minutes of inquiry. Sixty-one participants completed two thirds of the sessions or more, and 65 individuals participated in a waiting-list (WL) control group. Self-compassion, anxiety, depression and stress levels were assessed. The analysis of pre-post results suggests that the interventions increased self-compassion levels and decreased anxiety, depression and stress levels, whereas the WL group did not show any significant changes. The emotional changes in the intervention group were associated with the increase in self-compassion. However, at follow-up, the scores of emotional distress variables returned to the initial pre-intervention scores. These data can be interpreted in line with previous results that have shown the efficacy of self-compassion-based intervention programs. Given that this efficacy was not maintained at follow-up, data are discussed according to the pervasive role of a highly stressful context and-as described in other studies-the need for regular practice to maintain the benefits obtained.

This study relates emotional regulation strategies with dispositional mindfulness and the mediating role of time perspective. It is based on the fact that one of the mechanisms of mindfulness consists in providing protective emotional regulation strategies. At the same time, a direct relationship between dispositional mindfulness and time perspective has been observed. To do this, a representative sample of 320 Chilean adolescents from the city of Talcahuano, whose age ranged between 14 and 17 years old, and who were attending high school, was evaluated. The Zimbardo Time Perspective Inventory, the Difficulties in Emotion Regulation Scale, and the Five Facet Mindfulness Questionnaire were applied. Regression analysis results verified the close relationship between emotional regulation and dispositional mindfulness (R 2 = 0.54), as well as with the factors of time perspective (R 2 = 0.41), explaining, between both of them, 60% of the variance of difficulties in emotional regulation. The possible mediational role of time perspective between dispositional mindfulness and emotional regulation is established.

Human development implies deep changes in cognitive, attentional, emotional, and behavioral skills. Therefore, Mindfulness-Based Interventions (MBIs) should be adapted in terms of dose, frequency, kind of exercises, assessment methods, and expected effects regarding the abilities and limitations of each developmental period. The present review seeks to describe and compare MBIs characteristics, assessment methods, and effects in youth between 3 and 18 years old considering four developmental periods. A systematic review was carried out including experimental primary studies published during the last five years. Results show that the frequency of the sessions and program duration varies widely. Differences were observed in instructors' training and in assessment strategies. Discrepancies were observed regarding the effects of MBIs both within and between periods in cognitive, socio-emotional, symptoms, and mindfulness variables. Consistency was observed in prosocial behaviors for preschoolers, and in emotional and behavioral problems and hyperactivity in ages between preschool and early adolescence. Nevertheless, it was impossible to compare most results and determine consistency or discrepancy due to the lack of studies. Regarding mindfulness, it is defined and assessed in different ways in each period. Orientations are suggested to move from a compartmentalized view of isolated MBIs, towards an integrative perspective that allows tracing developmental trajectories for mindfulness and other key cognitive and socioemotional skills for children and adolescents.

Background: The aims of the study were to establish an adjustment model to analyze the relationship among mindfulness, balanced time perspective (BTP) and psychological well-being (PWB) in college students and to explore gender differences among the variables., Method: The sample consisted of 380 college students, 220 women and 160 men, uniformly distributed according to the university's faculties., Results: The results indicate that the synergy between mindfulness and BTP predicts the variance of PWB by 55%. Regarding gender differences, it was found that women have a greater tendency towards Past Positive than men and men a higher tendency towards Present Hedonistic than women. In addition, in the group of women, a stronger relationship was found among the variables and, consequently, a greater predictive value for PWB (58%), displaying an enhanced disposition to high PWB compared to men., Conclusions: Together, mindfulness and BTP promote optimal psychological functioning and alleviate or reduce discomfort. Thus, their promotion and training in universities is especially important given the high prevalence of anxiety and depressive symptoms in college students.

Ligand-independent epidermal growth factor receptor (EGFR) endocytosis is inducible by a variety of stress conditions converging upon p38 kinase. A less known pathway involves phosphatidic acid (PA) signaling toward the activation of type 4 phosphodiesterases (PDE4) that decrease cAMP levels and protein kinase A (PKA) activity. This PA/PDE4/PKA pathway is triggered with propranolol used to inhibit PA hydrolysis and induces clathrin-dependent and clathrin-independent endocytosis, followed by reversible accumulation of EGFR in recycling endosomes. Here we give further evidence of this signaling pathway using biosensors of PA, cAMP, and PKA in live cells and then show that it activates p38 and ERK1/2 downstream the PKA inhibition. Clathrin-silencing and IN/SUR experiments involved the activity of p38 in the clathrin-dependent route, while ERK1/2 mediates clathrin-independent EGFR endocytosis. The PA/PDE4/PKA pathway selectively increases the EGFR endocytic rate without affecting LDLR and TfR constitute endocytosis. This selectiveness is probably because of EGFR phosphorylation, as detected in Th1046/1047 and Ser669 residues. The EGFR accumulates at perinuclear recycling endosomes colocalizing with TfR, fluorescent transferrin, and Rab11, while a small proportion distributes to Alix-endosomes. A non-selective recycling arrest includes LDLR and TfR in a reversible manner. The PA/PDE4/PKA pathway involving both p38 and ERK1/2 expands the possibilities of EGFR transmodulation and interference in cancer., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Cancer therapy may be improved by the simultaneous interference of two or more oncogenic pathways contributing to tumor progression and aggressiveness, such as EGFR and p53. Tumor cells expressing gain-of-function (GOF) mutants of p53 (mutp53) are usually resistant to EGFR inhibitors and display invasive migration and AKT-mediated survival associated with enhanced EGFR recycling. D-Propranolol (D-Prop), the non-beta blocker enantiomer of propranolol, was previously shown to induce EGFR internalization through a PKA inhibitory pathway that blocks the recycling of the receptor. Here, we first show that D-Prop decreases the levels of EGFR at the surface of GOF mutp53 cells, relocating the receptor towards recycling endosomes, both in the absence of ligand and during stimulation with high concentrations of EGF or TGF-α. D-Prop also inactivates AKT signaling and reduces the invasive migration and viability of these mutp53 cells. Unexpectedly, mutp53 protein, which is stabilized by interaction with the chaperone HSP90 and mediates cell oncogenic addiction, becomes destabilized after D-Prop treatment. HSP90 phosphorylation by PKA and its interaction with mutp53 are decreased by D-Prop, releasing mutp53 towards proteasomal degradation. Furthermore, a single daily dose of D-Prop reproduces most of these effects in xenografts of aggressive gallbladder cancerous G-415 cells expressing GOF R282W mutp53, resulting in reduced tumor growth and extended mice survival. D-Prop then emerges as an old drug endowed with a novel therapeutic potential against EGFR- and mutp53-driven tumor traits that are common to a large variety of cancers.

Intimate relationship conflicts in young people are crucial experiences for change. They can lead to more or less satisfactory relationships, depending on individuals' skills to cope with these conflicts. This may or may not lead to violence in couples. Acceptance and self-regulation processes are an effective strategy to address individual factors such as avoidance and anxiety in intimate relationships of people in these age groups, thus preventing violence. The aim of this study was to assess the effects of an eight-session mindfulness and acceptance-based program (MAP). Participants (n = 40), who were aged from 18 to 25 years old, were randomly assigned to a group receiving the MAP or an active control group. Outcome measures were anxiety about abandonment, intimacy avoidance (Experiences in Close Relationships scale), well-being (Psychological Well-being Scale), dispositional mindfulness (Five Facet Mindfulness Questionnaire) and flexibility (Acceptance and Action Questionnaire-II). Measures were taken at pre-intervention, post-intervention and follow-up. Results showed that the MAP decreased anxiety ( p = 0.025) and avoidance ( p = 0.01) and increased mindfulness ( p < 0.001) and flexibility ( p = 0.001). In general, these improvements persisted at follow-up. Results are discussed in relation to the usefulness of mindfulness-acceptance strategies to cope with non-pathological intimate relationship conflicts.

Background and Objective: During cyclosporine-induced gingival overgrowth, the homeostatic balance of gingival connective tissue is disrupted leading to fibrosis. Galectins are glycan-binding proteins that can modulate a variety of cellular processes including fibrosis in several organs. Here, we study the role of galectin-8 (Gal-8) in the response of gingival connective tissue cells to cyclosporine., Methods: We used human gingival fibroblasts and mouse NIH3T3 cells treated with recombinant Gal-8 and/or cyclosporine for analyzing specific mRNA and protein levels through immunoblot, real-time polymerase chain reaction, ELISA and immunofluorescence, pull-down with Gal-8-Sepharose for Gal-8-to-cell surface glycoprotein interactions, short hairpin RNA for Gal-8 silencing and Student's t test and ANOVA for statistical analysis., Results: Galectin-8 stimulated type I collagen and fibronectin protein levels and potentiated CTGF protein levels in TGF-β1-stimulated human gingival fibroblasts. Gal-8 interacted with α5β1-integrin and type II TGF-β receptor. Gal-8 stimulated fibronectin protein and mRNA levels, and this response was dependent on FAK activity but not Smad2/3 signaling. Cyclosporine and tumor necrosis factor alpha (TNF-α) increased Gal-8 protein levels. Finally, silencing of galectin-8 in NIH3T3 cells abolished cyclosporine-induced fibronectin protein levels., Conclusion: Taken together, these results reveal for the first time Gal-8 as a fibrogenic stimulus exerted through β1-integrin/FAK pathways in human gingival fibroblasts, which can be triggered by cyclosporine. Further studies should explore the involvement of Gal-8 in human gingival tissues and its role in drug-induced gingival overgrowth., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: This study analyzes the relationship between depression and anxiety levels and positive and negative affect, self-esteem, and perceived social support from family and friends in an early and middle adolescent sample. These are psychological variables that are often associated with the prediction of emotional disorders, especially depression. Methods: Participants ( N = 467) were a representative sample of this group of adolescents and were recruited from schools in the city of Concepción, Chile. Part of the sample ( N = 177) was assessed three additional times-at one-, two-, and four-month intervals. Results: Results showed a practical stability of all measures across the four intervals, with no significant differences between sexes. Anxiety and depression displayed a similar pattern of significant relationships with affectivity, self-esteem, and social support. Depression had a higher correlation coefficient (-0.47) with positive affect, and so did anxiety with negative affect (0.58). Conclusions: Taking into account 23 initial scores on affectivity, self-esteem, and social support in predicting both depression and anxiety scores at one-month, two-month, and four-month intervals, positive affect was present in three regression analyses, predicting depression scores; negative affect was present in anxiety scores. Results are discussed according to previous findings, as well as the tripartite model.

Objective: There has been a growing interest in the study of the effectiveness of mindfulness-based interventions (MBIs). Many clinical trials and experimental designs have been implemented, with different samples and diverse MBI procedures. Reviews have shown unclear results, apart from a tendency to identify low-to-moderate effectiveness. The purpose of this review is to examine the effectiveness of MBIs on anxiety complaints, analyzing available systematic reviews and meta-analyses., Method: The literature search was done in MEDLINE (PubMed) and PsycINFO, from the first available review in 2003 until March 2020. From 82 initial references, 12 reviews were selected., Results: Reviews confirmed a moderate effect size of MBIs in improving anxiety symptoms. This efficacy was similar to that of well-established therapies for reducing anxiety symptoms, such as cognitive behavioral therapies. A large effect size was found when well-developed MBI protocols were applied., Discussion: More refined clinical trials are needed to establish clear conditions of MBI effectiveness (protocols, samples, psychological mechanisms, etc.). In addition, considering mindfulness processes, new outcome measures are needed (such as acceptance, self-awareness, or well-being) to test the incremental value of MBIs.

Galectin-8 (Gal-8) is a glycan-binding protein that modulates a variety of cellular processes interacting with cell surface glycoproteins. Neutralizing anti-Gal-8 antibodies that block Gal-8 functions have been described in autoimmune and inflammatory disorders, likely playing pathogenic roles. In the brain, Gal-8 is highly expressed in the choroid plexus and accordingly has been detected in human cerebrospinal fluid. It protects against central nervous system autoimmune damage through its immune-suppressive potential. Whether Gal-8 plays a direct role upon neurons remains unknown. Here, we show that Gal-8 protects hippocampal neurons in primary culture against damaging conditions such as nutrient deprivation, glutamate-induced excitotoxicity, hydrogen peroxide (H 2 O 2 )-induced oxidative stress, and β-amyloid oligomers (Aβo). This protective action is manifested even after 2 h of exposure to the harmful condition. Pull-down assays demonstrate binding of Gal-8 to selected β1-integrins, including α3 and α5β1. Furthermore, Gal-8 activates β1-integrins, ERK1/2, and PI3K/AKT signaling pathways that mediate neuroprotection. Hippocampal neurons in primary culture produce and secrete Gal-8, and their survival decreases upon incubation with human function-blocking Gal-8 autoantibodies obtained from lupus patients. Despite the low levels of Gal-8 expression detected by real-time PCR in hippocampus, compared with other brain regions, the complete lack of Gal-8 in Gal-8 KO mice determines higher levels of apoptosis upon H 2 O 2 stereotaxic injection in this region. Therefore, endogenous Gal-8 likely contributes to generate a neuroprotective environment in the brain, which might be eventually counteracted by human function-blocking autoantibodies.

Reflecting on negative emotional experiences can be adaptive but it can also maintain or intensify detrimental emotional states. Which factors determine whether reflection can have one consequence or another is unclear. This study focused on two research programs that have concentrated on this topic in the last decades: processing-mode theory (PMT) and self-distancing theory (SDT). The article described and contrasted both programs and their findings. The promising results that PMT and SDT have achieved in identifying the differences between the forms of adaptive and maladaptive reflection are highlighted. Likewise, the disconcerting contradictions observed between both programs that make integrating the findings difficult are indicated. The PMT states that adaptive reflection is concrete, and it is focused on the how of the experience. The SDT states that adaptive reflection is self-distanced and focused on the global meaning of the experience. The article finishes by indicating possible explanations for these apparent contradictions and outlines the challenges to be solved to improve comprehension of the topic.

The permeability of endothelial cells is regulated by the stability of the adherens junctions, which is highly sensitive to kinase-mediated phosphorylation and endothelial nitric oxide synthase (eNOS)-mediated S-nitrosylation of its protein components. Solid tumors can produce a variety of factors that stimulate these signaling pathways leading to endothelial cell hyperpermeability. This generates stromal conditions that facilitate tumoral growth and dissemination. Galectin-8 (Gal-8) is overexpressed in several carcinomas and has a variety of cellular effects that can contribute to tumor pathogenicity, including angiogenesis. Here we explored whether Gal-8 has also a role in endothelial permeability. We show that recombinant Gal-8 activates eNOS, induces S-nitrosylation of p120-catenin (p120) and dissociation of adherens junction, leading to hyperpermeability of the human endothelial cell line EAhy926. This pathway involves focal-adhesion kinase (FAK) activation downstream of eNOS as a requirement for eNOS-mediated p120 S-nitrosylation. This suggests a reciprocal, yet little understood, regulation of phosphorylation and S-nitrosylation events acting upon adherens junction permeability. In addition, glutathione S-transferase (GST)-Gal-8 pull-down experiments and function-blocking β1-integrin antibodies point to β1-integrins as cell surface components involved in Gal-8-induced hyperpermeability. Endogenous Gal-8 secreted from the breast cancer cell line MCF-7 has similar hyperpermeability and signaling effects. Furthermore, the mouse cremaster model system showed that Gal-8 also activates eNOS, induces S-nitrosylation of adherens junction components and is an effective hyperpermeability agent in vivo. These results add endothelial permeability regulation by S-nitrosylation as a new function of Gal-8 that can potentially contribute to the pathogenicity of tumors overexpressing this lectin., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Complete activation of B cells relies on their capacity to extract tethered antigens from immune synapses by either exerting mechanical forces or promoting their proteolytic degradation through lysosome secretion. Whether antigen extraction can also be tuned by local cues originating from the lymphoid microenvironment has not been investigated. We here show that the expression of Galectin-8-a glycan-binding protein found in the extracellular milieu, which regulates interactions between cells and matrix proteins-is increased within lymph nodes under inflammatory conditions where it enhances B cell arrest phases upon antigen recognition in vivo and promotes synapse formation during BCR recognition of immobilized antigens. Galectin-8 triggers a faster recruitment and secretion of lysosomes toward the B cell-antigen contact site, resulting in efficient extraction of immobilized antigens through a proteolytic mechanism. Thus, extracellular cues can determine how B cells sense and extract tethered antigens and thereby tune B cell responses in vivo., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

Epithelial cells can acquire invasive and tumorigenic capabilities through epithelial-mesenchymal-transition (EMT). The glycan-binding protein galectin-8 (Gal-8) activates selective β1-integrins involved in EMT and is overexpressed by certain carcinomas. Here we show that Gal-8 overexpression or exogenous addition promotes proliferation, migration, and invasion in nontumoral Madin-Darby canine kidney (MDCK) cells, involving focal-adhesion kinase (FAK)-mediated transactivation of the epidermal growth factor receptor (EGFR), likely triggered by α5β1integrin binding. Under subconfluent conditions, Gal-8-overexpressing MDCK cells (MDCK-Gal-8 H ) display hallmarks of EMT, including decreased E-cadherin and up-regulated expression of vimentin, fibronectin, and Snail, as well as increased β-catenin activity. Changes related to migration/invasion included higher expression of α5β1 integrin, extracellular matrix-degrading MMP13 and urokinase plasminogen activator/urokinase plasminogen activator receptor (uPA/uPAR) protease systems. Gal-8-stimulated FAK/EGFR pathway leads to proteasome overactivity characteristic of cancer cells. Yet MDCK-Gal-8 H cells still develop apical/basolateral polarity reverting EMT markers and proteasome activity under confluence. This is due to the opposite segregation of Gal-8 secretion (apical) and β1-integrins distribution (basolateral). Strikingly, MDCK-Gal-8 H cells acquired tumorigenic potential, as reflected in anchorage-independent growth in soft agar and tumor generation in immunodeficient NSG mice. Therefore, Gal-8 can promote oncogenic-like transformation of epithelial cells through partial and reversible EMT, accompanied by higher proliferation, migration/invasion, and tumorigenic properties., (© 2018 Oyanadel et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

Galectin-8 (Gal-8) is a member of a glycan-binding protein family that regulates the immune system, among other functions, and is a target of antibodies in autoimmune disorders. However, its role in multiple sclerosis (MS), an autoimmune inflammatory disease of the central nervous system (CNS), remains unknown. We study the consequences of Gal-8 silencing on lymphocyte subpopulations and the development of experimental autoimmune encephalitis (EAE), to then assess the presence and clinical meaning of anti-Gal-8 antibodies in MS patients. Lgals8/Lac-Z knock-in mice lacking Gal-8 expression have higher polarization toward Th17 cells accompanied with decreased CCR6+ and higher CXCR3+ regulatory T cells (Tregs) frequency. These conditions result in exacerbated MOG35-55 peptide-induced EAE. Gal-8 eliminates activated Th17 but not Th1 cells by apoptosis and ameliorates EAE in C57BL/6 wild-type mice. β-gal histochemistry reflecting the activity of the Gal-8 promoter revealed Gal-8 expression in a wide range of CNS regions, including high expression in the choroid-plexus. Accordingly, we detected Gal-8 in human cerebrospinal fluid, suggesting a role in the CNS immune-surveillance circuit. In addition, we show that MS patients generate function-blocking anti-Gal-8 antibodies with pathogenic potential. Such antibodies block cell adhesion and Gal-8-induced Th17 apoptosis. Furthermore, circulating anti-Gal-8 antibodies associate with relapsing-remitting MS (RRMS), and not with progressive MS phenotypes, predicting clinical disability at diagnosis within the first year of follow-up. Our results reveal that Gal-8 has an immunosuppressive protective role against autoimmune CNS inflammation, modulating the balance of Th17 and Th1 polarization and their respective Tregs. Such a role can be counteracted during RRMS by anti-Gal-8 antibodies, worsening disease prognosis. Even though anti-Gal-8 antibodies are not specific for MS, our results suggest that they could be a potential early severity biomarker in RRMS.

The interaction between acute myeloid leukemia cells (AML) with the bone marrow stroma cells (BMSCs) determines a protective environment that favors tumor development and resistance to conventional chemotherapy. We showed that BMSCs secrete soluble factors that protect AML cells from Ara-C induced cytotoxicity. This leukemia chemoresistance is associated with a decrease in the equilibrative nucleoside transporter (ENT1) activity by inducing removal of ENT1 from the cell surface. Reduction of cell proliferation was also observed with activation of AKT and mTOR-dependent cell survival pathways, which may also contribute to the tumor chemoprotection. Analysis of primary BMSC cultures has demonstrated that AML patients with stroma capable to confer Ara-C resistance in vitro compared to AML patients without this stroma capacity were associated with a worse prognosis. The two year overall survival rate was 0% versus 80% respectively (p=0.0001). This is the first report of a chemoprotection mechanism based on the removal of a drug transporter from the cell surface and most importantly the first time that a stroma phenotype has correlated with prognostic outcome in cancer.

Background: Glioblastoma is one of the most aggressive cancers of the brain. Malignant traits of glioblastoma cells include elevated migration, proliferation and survival capabilities. Galectins are unconventionally secreted glycan-binding proteins that modulate processes of cell adhesion, migration, proliferation and apoptosis by interacting with beta-galactosides of cell surface glycoproteins and the extracellular matrix. Galectin-8 is one of the galectins highly expressed in glioblastoma cells. It has a unique selectivity for terminally sialylated glycans recently found enhanced in these highly malignant cells. A previous study in glioblastoma cell lines reported that Gal-8 coating a plastic surface stimulates two-dimensional motility. Because in other cells Gal-8 arrests proliferation and induces apoptosis, here we extend its study by analyzing all of these processes in a U87 glioblastoma cell model., Methods: We used immunoblot and RT-PCR for Gal-8 expression analysis, recombinant Gal-8 produced in a bacteria system for Gal-8 treatment of the cells, and shRNA in lentivirus transduction for Gal-8 silencing. Cell migration as assessed in transwell filters. Cell proliferation, cell cycle and apoptosis were analyzed by FACS., Results: Gal-8 as a soluble stimulus triggered chemotactic migration of U87 cells across the polycarbonate filter of transwell chambers, almost as intensively as fetal bovine serum. Unexpectedly, Gal-8 also enhanced U87 cell growth. Co-incubation of Gal-8 with lactose, which blocks galectin-glycan interactions, abrogated both effects. Immunoblot showed Gal-8 in conditioned media reflecting its secretion. U87 cells transduced with silencing shRNA in a lentiviral vector expressed and secreted 30-40 % of their normal Gal-8 levels. These cells maintained their migratory capabilities, but decreased their proliferation rate and underwent higher levels of apoptosis, as revealed by flow cytometry analysis of cell cycle, CFSE and activated caspase-3 staining. Proliferation seemed to be more sensitive than migration to Gal-8 expression levels., Conclusions: Gal-8, either secreted or exogenously enriched in the media, and acting through extracellular glycan interactions, constitutes a strong stimulus of directional migration in glioblastoma U87 cells and for the first time emerges as a factor that promotes proliferation and prevents apoptosis in cancerous cells. These properties could potentially contribute to the exaggerated malignancy of glioblastoma cells.

Introduction: The study of time perception and mental illness has given priority to time estimation over time perspective. Considering Zimbardo’s theory on five dimensions of time perspective, and balanced time perspective profile, this study has aimed to compare people with severe mental illness (SMI) and healthy people, with measurements of time perspective and time estimation and to assess whether the time perspective profile influences the quality of life in people with SMI., Material and Methods: Using a quasi-experimental design, a clinical group (n=167) corresponding to four samples of severe mental disorders (major depression, bipolar disorder, schizophrenia and personality disorders) and healthy people (n=167) were compared in their performance regarding time perspective and time estimation. After, the clinical sample was grouped according to their deviation from the balanced time perspective profile (DBTP) and negative profile (DNTP). These groups were evaluated with health measures and time estimation tasks., Results and Conclusion: Through the ANOVA, it can be seen that the time perspective profile affects health measurements. There are significant differences between the clinical sample and controls regarding time perspective and time estimation. Within the group of patients, it was observed that those who were closer to the BTP profile had better physical health, and less hopelessness (p<.05). This measurement may favor interventions related to a balanced profile. Results are discussed in relation to contribution of time perspective in the assessment, treatment and quality of life of people with SMI.

Epidermal growth factor receptor (EGFR) exaggerated (oncogenic) function is currently targeted in cancer treatment with drugs that block receptor ligand binding or tyrosine kinase activity. Because endocytic trafficking is a crucial regulator of EGFR function, its pharmacological perturbation might provide a new anti-tumoral strategy. Inhibition of phosphatidic acid (PA) phosphohydrolase (PAP) activity has been shown to trigger PA signaling towards type 4 phosphodiesterase (PDE4) activation and protein kinase A inhibition, leading to internalization of empty/inactive EGFR. Here, we used propranolol, its l- and d- isomers and desipramine as PAP inhibitors to further explore the effects of PAP inhibition on EGFR endocytic trafficking and its consequences on EGFR-dependent cancer cell line models. PAP inhibition not only made EGFR inaccessible to stimuli but also prolonged the signaling lifetime of ligand-activated EGFR in recycling endosomes. Strikingly, such endocytic perturbations applied in acute/intermittent PAP inhibitor treatments selectively impaired cell proliferation/viability sustained by an exaggerated EGFR function. Phospholipase D inhibition with FIPI (5-fluoro-2-indolyl des-chlorohalopemide) and PDE4 inhibition with rolipram abrogated both the anti-tumoral and endocytic effects of PAP inhibition. Prolonged treatments with a low concentration of PAP inhibitors, although without detectable endocytic effects, still counteracted cell proliferation, induced apoptosis and decreased anchorage-independent growth of cells bearing EGFR oncogenic influences. Overall, our results show that PAP inhibitors can counteract EGFR oncogenic traits, including receptor overexpression or activating mutations resistant to current tyrosine kinase inhibitors, perturbing EGFR endocytic trafficking and perhaps other as yet unknown processes, depending on treatment conditions. This puts PAP activity forward as a new suitable target against EGFR-driven malignancy., (© 2014 FEBS.)

Sonic Hedgehog (Shh/GLI) and EGFR signaling pathways modulate Neural Stem Cell (NSC) proliferation. How these signals cooperate is therefore critical for understanding normal brain development and function. Here we report a novel acute effect of Shh signaling on EGFR function. We show that during late neocortex development, Shh mediates the activation of the ERK1/2 signaling pathway in Radial Glial cells (RGC) through EGFR transactivation. This process is dependent on metalloprotease activity and accounts for almost 50% of the EGFR-dependent mitogenic response of late NSCs. Furthermore, in HeLa cancer cells, a well-known model for studying the EGFR receptor function, Shh also induces cell proliferation involving EGFR activation, as reflected by EGFR internalization and ERK1/2 phosphorylation. These findings may have important implications for understanding the mechanisms that regulate NSC proliferation during neurogenesis and may lead to novel approaches to the treatment of tumors.

Galectin-8 belongs to a family of mammalian lectins that recognize glycoconjugates present on different cell surface components and modulate a variety of cellular processes. A role of Gal-8 in the immune system has been proposed based on its effects in immune cells, including T and B lymphocytes, as well as the presence of anti-Gal-8 autoantibodies in the prototypic autoimmune disease systemic lupus erythematosus (SLE). We have previously described that Gal-8 induces apoptosis in activated T cells interacting with certain β1 integrins and this effect is counteracted by the anti-Gal-8 autoantibodies. Given that Gal-8 can potentially interact with several glycoproteins, here we analyzed the β2 integrin Lymphocyte Function-Associated Antigen-1 (LFA-1), which is involved in leukocyte cell adhesion and immunological synapses. We show by GST-pull down assays that Gal-8 interacts with LFA-1 and this interaction is inhibited by anti-Gal-8 autoantibodies isolated from SLE patients. In cell adhesion assays, Gal-8 precluded the interaction of LFA-1 with its ligand Intracellular Adhesion Molecule-1 (ICAM-1). These results suggest that Gal-8 can exert immunosuppressive action not only by inducing apoptosis in activated T cells but also by negatively modulating the crucial function of LFA-1 in the immune system, while function-blocking autoantibodies counteract these effects.

Galectins have been implicated in T cell homeostasis playing complementary pro-apoptotic roles. Here we show that galectin-8 (Gal-8) is a potent pro-apoptotic agent in Jurkat T cells inducing a complex phospholipase D/phosphatidic acid signaling pathway that has not been reported for any galectin before. Gal-8 increases phosphatidic signaling, which enhances the activity of both ERK1/2 and type 4 phosphodiesterases (PDE4), with a subsequent decrease in basal protein kinase A activity. Strikingly, rolipram inhibition of PDE4 decreases ERK1/2 activity. Thus Gal-8-induced PDE4 activation releases a negative influence of cAMP/protein kinase A on ERK1/2. The resulting strong ERK1/2 activation leads to expression of the death factor Fas ligand and caspase-mediated apoptosis. Several conditions that decrease ERK1/2 activity also decrease apoptosis, such as anti-Fas ligand blocking antibodies. In addition, experiments with freshly isolated human peripheral blood mononuclear cells, previously stimulated with anti-CD3 and anti-CD28, show that Gal-8 is pro-apoptotic on activated T cells, most likely on a subpopulation of them. Anti-Gal-8 autoantibodies from patients with systemic lupus erythematosus block the apoptotic effect of Gal-8. These results implicate Gal-8 as a novel T cell suppressive factor, which can be counterbalanced by function-blocking autoantibodies in autoimmunity.

The epithelial-specific adaptor AP1B sorts basolateral proteins, but the trafficking routes where it performs its sorting role remain controversial. Here, we used an RNAi approach to knock down the medium subunit of AP1B (mu1B) in the prototype epithelial cell line Madin-Darby canine kidney (MDCK). Mu1B-knocked down MDCK cells displayed loss of polarity of several endogenous and exogenous basolateral markers transduced via adenovirus vectors, but exhibited normal polarity of apical markers. We chose two well characterized basolateral protein markers, the transferrin receptor (TfR) and the vesicular stomatitis virus G protein, to study the sorting role of AP1B. A surface-capture assay introduced here showed that mu1B-knocked down MDCK cells plated on filters at confluency and cultured for 4.5 d, sorted TfR correctly in the biosynthetic route but incorrectly in the recycling route. In contrast, these same cells missorted vesicular stomatitis virus G apically in the biosynthetic route. Strikingly, recently confluent MDCK cells (1-3 d) displayed AP1B-dependence in the biosynthetic route of TfR, which decreased with additional days in culture. Sucrose density gradient analysis detected AP1B predominantly in TfR-rich endosomal fractions in MDCK cells confluent for 1 and 4 d. Our results are consistent with the following model: AP1B sorts basolateral proteins in both biosynthetic and recycling routes of MDCK cells, as a result of its predominant functional localization in recycling endosomes, which constitute a post-Golgi station in the biosynthetic route of some plasma membrane proteins. TfR utilizes a direct route from Golgi to basolateral membrane that is established as the epithelial monolayer matures.

Integrin-mediated encounters of T cells with extracellular cues lead these cells to adhere to a variety of substrates and acquire a spread phenotype needed for their tissue incursions. We studied the effects of galectin-8 (Gal-8), a beta-galactoside binding lectin, on Jurkat T cells. Immobilized Gal-8 bound alpha1beta1, alpha3beta1 and alpha5beta1 but not alpha2beta1 and alpha4beta1 and adhered these cells with similar kinetics to immobilized fibronectin (FN). Function-blocking experiments with monoclonal anti-integrin antibodies suggested that alpha5beta1 is the main mediator of cell adhesion to this lectin. Gal-8, but not FN, induced extensive cell spreading frequently leading to a polarized phenotype characterized by an asymmetric lamellipodial protrusion. These morphological changes involved actin cytoskeletal rearrangements controlled by PI3K, Rac-1 and ERK1/2 activity. Gal-8-induced Rac-1 activation and binding to alpha1 and alpha5 integrins have not been described in any other cellular system. Strikingly, Gal-8 was also a strong stimulus on Jurkat cells in suspension, triggering ERK1/2 activation that in most adherent cells is instead dependent on cell attachment. In addition, we found that patients with systemic lupus erythematosus (SLE), a prototypic autoimmune disorder, produce Gal-8 autoantibodies that impede both its binding to integrins and cell adhesion. These are the first function-blocking autoantibodies reported for a member of the galectin family. These results indicate that Gal-8 constitutes a novel extracellular stimulus for T cells, able to bind specific beta1 integrins and to trigger signaling pathways conducive to cell spreading. Gal-8 could modulate a wide range of T cell-driven immune processes that eventually become altered in autoimmune disorders.