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Synapsin I (SynI) is a synaptic vesicle (SV)-associated phosphoprotein that modulates neurotransmission by controlling SV trafficking. The SynI C-domain contains a highly conserved ATP binding site mediating SynI oligomerization and SV clustering and an adjacent main Ca 2+ binding site, whose physiological role is unexplored. Molecular dynamics simulations revealed that the E373K point mutation irreversibly deletes Ca 2+ binding to SynI, still allowing ATP binding, but inducing a destabilization of the SynI oligomerization interface. Here, we analyzed the effects of this mutation on neurotransmitter release and short-term plasticity in excitatory and inhibitory synapses from primary hippocampal neurons. Patch-clamp recordings showed an increase in the frequency of miniature excitatory postsynaptic currents (EPSCs) that was totally occluded by exogenous Ca 2+ chelators and associated with a constitutive increase in resting terminal Ca 2+ concentrations. Evoked EPSC amplitude was also reduced, due to a decreased readily releasable pool (RRP) size. Moreover, in both excitatory and inhibitory synapses, we observed a marked impaired recovery from synaptic depression, associated with impaired RRP refilling and depletion of the recycling pool of SVs. Our study identifies SynI as a novel Ca 2+ buffer in excitatory terminals. Blocking Ca 2+ binding to SynI results in higher constitutive Ca 2+ levels that increase the probability of spontaneous release and disperse SVs. This causes a decreased size of the RRP and an impaired recovery from depression due to the failure of SV reclustering after sustained high-frequency stimulation. The results indicate a physiological role of Ca 2+ binding to SynI in the regulation of SV clustering and trafficking in nerve terminals., (© 2022. The Author(s).)

Objectives: This study investigated the role of ethnic/racial composition in schools and neighborhoods in (a) predicting family cultural socialization and (b) moderating the relation between family cultural socialization and young children's social competence over time., Method: Two nationally representative, longitudinal samples were used from the Early Childhood Longitudinal Study, Kindergarten Class of 1998-99 and 2010-11 cohorts. The analytic sample included 11,870 ethnic/racial minority children (mean age was 5.66 years old at Wave 1; 50% female; 31% Black, 49% Latinx, 18% Asian American, 2% Native American)., Results: Path analyses showed that families practiced more cultural socialization in more diverse schools and neighborhoods. Moreover, family cultural socialization was most beneficial for children's social competence when they were in diverse settings with few coethnics., Conclusions: The results highlighted cultural socialization as a tool that ethnic/racial minority families use to help their children navigate ethnic/racial diversity and numeric marginalization in social settings. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Objective: To describe the approach and outcomes from two cancer centres in Southern and Northern Europe during the first wave of coronavirus disease 2019 (COVID-19) of patients with head and neck cancer (HNC)., Methods: Data collection was performed on a retrospective cohort of patients surgically treated for primary HNC between March and May 2020, using data from two tertiary hospitals: the European Institute of Oncology (Milan) and Guy's & St Thomas' NHS Foundation Trust (London)., Results: We included 77 patients with HNC. More patients with COVID-19 were taking angiotensin-converting enzyme (ACE) inhibitors and had Clavien-Dindo Classification grade I compared to negative patients, respectively (60% vs 22% [ p = 0.058] and 40% vs 8% [ p = 0.025]). Multivariate logistic regression analyses confirmed our data ( p = 0.05 and 0.03, respectively). Sex and age were statistically significantly different ( p = 0.05 and <0.001 respectively), showing more male patients (75% vs 53.66%, respectively) and more elderly patients in Italy than in the United Kingdom (patients aged >63 years: 69.44% vs 29.27%)., Conclusions: This study presents a large cohort of patients with HNC with nasopharyngeal swab during the first peak of the COVID-19 pandemic in Europe. Patients with HNC with COVID-19 appeared more likely to develop postsurgical complications and to be taking ACE inhibitors. The preventive measures adopted guaranteed the continuation of therapeutic surgical intervention.

While the detrimental consequences of racial/ethnic discrimination for adolescent well-being are well-established, less is known about the impact of SES-based discrimination and the potential protective benefits of adolescents' intraindividual assets. The current study addressed these gaps by investigating the longitudinal associations between racial/ethnic and SES-based educator-perpetrated discrimination and adolescents' academic well-being and assessed whether psychological resources moderated these pathways. To do so, the study used longitudinal data from a diverse sample of 750 9 th grade students (54% female; 41% White, 34% Latina/o/x, 8% Asian American, 6% African American, 11% biracial/other race/ethnicity; 43% had parents with an associate's degree or less) in the Southwestern U.S. who were subsequently surveyed one year later. Educator-perpetrated racial/ethnic discrimination was negatively associated with students' school engagement, and both psychological resilience and self-efficacy emerged as protective for students' educational expectations in the face of racial/ethnic and SES-based discrimination, respectively. The results of the current study highlight the role of discriminatory treatment in educational disparities and provide insights on effective coping strategies to combat the negative impacts of discrimination in academics., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Objective. In this work we adapted a protocol for the fast generation of human neurons to build 3D neuronal networks with controlled structure and cell composition suitable for systematic electrophysiological investigations. Approach. We used biocompatible chitosan microbeads as scaffold to build 3D networks and to ensure nutrients-medium exchange from the core of the structure to the external environment. We used excitatory neurons derived from human-induced pluripotent stem cells (hiPSCs) co-cultured with astrocytes. By adapting the well-established NgN2 differentiation protocol, we obtained 3D engineered networks with good control over cell density, volume and cell composition. We coupled the 3D neuronal networks to 60-channel micro electrode arrays (MEAs) to monitor and characterize their electrophysiological development. In parallel, we generated two-dimensional neuronal networks cultured on chitosan to compare the results of the two models. Main results. We sustained samples until 60 d in vitro (DIV) and 3D cultures were healthy and functional. From the structural point of view, the hiPSC derived neurons were able to adhere to chitosan microbeads and to form a stable 3D assembly thanks to the connections among cells. From a functional point of view, neuronal networks showed spontaneous activity after a couple of weeks. Significance. We presented a particular method to generate 3D engineered cultures for the first time with human-derived neurons coupled to MEAs, overcoming some of the limitations related to 2D and 3D neuronal networks and thus increasing the therapeutic target potential of these models for biomedical applications., (Creative Commons Attribution license.)

Background: The novel coronavirus disease of 2019 (COVID-19) outbreak provoked a profound healthcare system reorganization. This study aimed to compare the reasons for requesting a non-deferrable neurological evaluation before the COVID-19 pandemic and during the lockdown., Methods: Retrospective observational study including non-deferrable neurological outpatients before the pandemic (pre-COVID-19 group, n = 223) and during the Italian second wave of the COVID-19 pandemic (LOCKDOWN group, n = 318)., Results: The number of patients sent for cerebrovascular disorders, headache, and vertigo significantly dropped between the pre-COVID-19 era and the lockdown period. While in the pre-COVID-19 group, the most frequent diagnosis was cerebrovascular disorder; neuropsychiatric disorders ranked first in the LOCKDOWN group. Moreover, the percentage of appropriate non-deferrable neurological evaluations significantly increased in the LOCKDOWN group compared with the pre-COVID-19 group., Discussion: Our study shows a significant increase of neuropsychiatric disorders in non-deferrable neurologic evaluations during the Italian second wave of the COVID-19. Overall, cases were more severe and required a more complex management during the lockdown compared with the pre-COVID era. These findings confirm that a careful approach to prevent the psychological consequences of the pandemic is needed, and long-term rearrangements of the healthcare system are desirable to guarantee appropriate management.

The current study investigated how parents' and teachers' educational expectations both directly and indirectly shaped young people's academic outcomes in a nationally-representative sample of high school students (Education Longitudinal Study; N = 9654 adolescents). Higher parent and math teacher expectations in 10th grade were associated with better 12th grade math scores and higher grade point averages, math course-taking sequence, and educational attainment two years post-high school. High parent expectations generally magnified the particularly strong positive effects of high math teacher expectations, and there was some evidence of variation in links between adult expectations and outcomes by both student race/ethnicity and socioeconomic status. Parents' educational involvement at school, teacher-student relationships, and school-parent communication mediated the links between adult educational expectations and academic outcomes., (© 2021 Wiley Periodicals LLC.)

Educational interventions typically center on youth displaying early academic risk, potentially overlooking those falling off track academically later in their educational careers. The current study investigated the extent to which life course transitions experienced during adolescence were linked to falling off-track academically in high school. Data from the National Longitudinal Study of Adolescent to Adult Health (N = 4284; 53% female; M age  = 14.88) documented that 1516 students displayed no educational risk in early high school, yet 14% did not pursue 4-year college by age 24. Analyses revealed the unique life course transitions predictive of falling off-track academically (i.e., sexual intercourse, alcohol use, family transitions, residential mobility). The study's findings highlight important intervention avenues to promote adolescents' continued educational persistence.

Mesenchymal stromal cells (MSCs) are currently used for cartilage cell therapy because of their well proven capacity to differentiate in chondrocytes. The advantage of MSC-based therapy is the possibility of producing a high number of chondrocytes for implants. The transplant procedure, however, has some limitations, since MSCs may produce non-functional chondrocytes. This limit has been challenged by cultivating MSC in media with hydrogels containing hyaluronic acid (HA), extractive chondroitin sulfate (CS), or bio-fermentative unsulphated chondroitin (BC) alone or in combination. Nevertheless, a clear study of the effect of glycosaminoglycans (GAGs) on chondrocyte differentiation is still lacking, especially for the newly obtained unsulfated chondroitin of biotechnological origin. Are these GAGs playing a role in the commitment of stem cells to chondrocyte progenitors and in the differentiation of progenitors to mature chondrocytes? Alternatively, do they have a role only in one of these biological processes? We evaluated the role of HA, CS, and - above all - BC in cell commitment and chondrocyte differentiation of MSCs by supplementing these GAGs in different phases of in vitro cultivation. Our data provided evidence that a combination of HA and CS or of HA and BC supplemented during the terminal in vitro differentiation and not during cell commitment of MSCs improved chondrocytes differentiation without the presence of fibrosis (reduced expression of Type I collagen). This result suggests that a careful evaluation of extracellular cues for chondrocyte differentiation is fundamental to obtaining a proper maturation process., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Alessio, Stellavato, Aprile, Cimini, Vassallo, Di Bernardo, Galderisi and Schiraldi.)

The cells present in the stromal compartment of many tissues are a heterogeneous population containing stem cells, progenitor cells, fibroblasts, and other stromal cells. A SSEA3(+) cell subpopulation isolated from human stromal compartments showed stem cell properties. These cells, known as multilineage-differentiating stress-enduring (MUSE) cells, are capable of resisting stress and possess an excellent ability to repair DNA damage. We isolated MUSE cells from different mouse stromal compartments, such as those present in bone marrow, subcutaneous white adipose tissue, and ear connective tissue. These cells showed overlapping in vitro biological properties. The mouse MUSE cells were positive for stemness markers such as SOX2, OCT3/4, and NANOG. They also expressed TERT, the catalytic telomerase subunit. The mouse MUSE cells showed spontaneous commitment to differentiation in meso/ecto/endodermal derivatives. The demonstration that multilineage stem cells can be isolated from an animal model, such as the mouse, could offer a valid alternative to the use of other stem cells for disease studies and envisage of cellular therapies.

During their life span, cells have two possible states: a non-cycling, quiescent state (G0) and a cycling, activated state. Cells may enter a reversible G0 state of quiescence or, alternatively, they may undergo an irreversible G0 state. The latter may be a physiological differentiation or, following a stress event, a senescent status. Discrimination among the several G0 states represents a significant investigation, since quiescence, differentiation, and senescence are progressive phenomena with intermediate transitional stages. We used the expression of Ki67, RPS6, and beta-galactosidase to identify healthy cells that progressively enter and leave quiescence through G0-entry, G0 and G0-alert states. We then evaluated how cells may enter senescence following a genotoxic stressful event. We identified an initial stress stage with the expression of beta-galactosidase and Ki67 proliferation marker. Cells may recover from stress events or become senescent passing through early and late senescence states. Discrimination between quiescence and senescence was based on the expression of RPS6, a marker of active protein synthesis that is present in senescent cells but absent in quiescent cells. Even taking into account that fixed G0 states do not exist, our molecular algorithm may represent a method for identifying turning points of G0 transitional states that continuously change.

Mutations in PRoline Rich Transmembrane protein 2 (PRRT2) cause pleiotropic syndromes including benign infantile epilepsy, paroxysmal kinesigenic dyskinesia, episodic ataxia, that share the paroxysmal character of the clinical manifestations. PRRT2 is a neuronal protein that plays multiple roles in the regulation of neuronal development, excitability, and neurotransmitter release. To better understand the physiopathology of these clinical phenotypes, we investigated PRRT2 interactome in mouse brain by a pulldown-based proteomic approach and identified α1 and α3 Na + /K + ATPase (NKA) pumps as major PRRT2-binding proteins. We confirmed PRRT2 and NKA interaction by biochemical approaches and showed their colocalization at neuronal plasma membrane. The acute or constitutive inactivation of PRRT2 had a functional impact on NKA. While PRRT2-deficiency did not modify NKA expression and surface exposure, it caused an increased clustering of α3-NKA on the plasma membrane. Electrophysiological recordings showed that PRRT2-deficiency in primary neurons impaired NKA function during neuronal stimulation without affecting pump activity under resting conditions. Both phenotypes were fully normalized by re-expression of PRRT2 in PRRT2-deficient neurons. In addition, the NKA-dependent afterhyperpolarization that follows high-frequency firing was also reduced in PRRT2-silenced neurons. Taken together, these results demonstrate that PRRT2 is a physiological modulator of NKA function and suggest that an impaired NKA activity contributes to the hyperexcitability phenotype caused by PRRT2 deficiency.

Muse cells are adult stem cells that are present in the stroma of several organs and possess an enduring capacity to cope with endogenous and exogenous genotoxic stress. In cell therapy, the peculiar biological properties of Muse cells render them a possible natural alternative to mesenchymal stromal cells (MSCs) or to in vitro-generated pluripotent stem cells (iPSCs). Indeed, some studies have proved that Muse cells can survive in adverse microenvironments, such as those present in damaged/injured tissues. We performed an evaluation of Muse cells' proteome under basic conditions and followed oxidative stress treatment in order to identify ontologies, pathways, and networks that can be related to their enduring stress capacity. We executed the same analysis on iPSCs and MSCs, as a comparison. The Muse cells are enriched in several ontologies and pathways, such as endosomal vacuolar trafficking related to stress response, ubiquitin and proteasome degradation, and reactive oxygen scavenging. In Muse cells, the protein-protein interacting network has two key nodes with a high connectivity degree and betweenness: NFKB and CRKL. The protein NFKB is an almost-ubiquitous transcription factor related to many biological processes and can also have a role in protecting cells from apoptosis during exposure to a variety of stressors. CRKL is an adaptor protein and constitutes an integral part of the stress-activated protein kinase (SAPK) pathway. The identified pathways and networks are all involved in the quality control of cell components and may explain the stress resistance of Muse cells.

Purpose: The purpose of our study was to assess the potential role of chest CT in the early detection of COVID-19 pneumonia and to explore its role in patient management in an adult Italian population admitted to the Emergency Department., Methods: Three hundred and fourteen patients presented with clinically suspected COVID-19, from March 3 to 23, 2020, were evaluated with PaO2/FIO2 ratio from arterial blood gas, RT-PCR assay from nasopharyngeal swab sample and chest CT. Patients were classified as COVID-19 negative and COVID-19 positive according to RT-PCR results, considered as a reference. Images were independently evaluated by two radiologists blinded to the RT-PCR results and classified as "CT positive" or "CT negative" for COVID-19, according to CT findings., Results: According to RT-PCR results, 152 patients were COVID-19 negative (48%) and 162 were COVID-19 positive (52%). We found substantial agreement between RT-PCR results and CT findings (p < 0.000001), as well as an almost perfect agreement between the two readers. Mixed GGO and consolidation pattern with peripheral and bilateral distribution, multifocal or diffuse abnormalities localized in both upper lung and lower lung, in association with interlobular septal thickening, bronchial wall thickening and air bronchogram, showed higher frequency in COVID-positive patients. We also found a significant correlation between CT findings and patient's oxygenation status expressed by PaO2/FIO2 ratio., Conclusion: Chest CT has a useful role in the early detection and in patient management of COVID-19 pneumonia in a pandemic. It helps in identifying suspected patients, cutting off the route of transmission and avoiding further spread of infection.

Background: The term mesenchymal stromal cells (MSCs) designates an assorted cell population comprised of stem cells, progenitor cells, fibroblasts, and stromal cells. MSCs contribute to the homeostatic maintenance of many organs through paracrine and long-distance signaling. Tissue environment, in both physiological and pathological conditions, may affect the intercellular communication of MSCs., Methods: We performed a secretome analysis of MSCs isolated from subcutaneous adipose tissue (sWAT) and visceral adipose tissue (vWAT), and from bone marrow (BM), of normal and obese mice., Results: The MSCs isolated from tissues of healthy mice share a common core of released factors: components of cytoskeletal and extracellular structures; regulators of basic cellular functions, such as protein synthesis and degradation; modulators of endoplasmic reticulum stress; and counteracting oxidative stress. It can be hypothesized that MSC secretome beneficially affects target cells by the horizontal transfer of many released factors. Each type of MSC may exert specific signaling functions, which could be determined by looking at the many factors that are exclusively released from every MSC type. The vWAT-MSCs release factors that play a role in detoxification activity in response to toxic substances and drugs. The sWAT-MSC secretome contains proteins involved in in chondrogenesis, osteogenesis, and angiogenesis. Analysis of BM-MSC secretome revealed that these cells exert a signaling function by remodeling extracellular matrix structures, such as those containing glycosaminoglycans. Obesity status profoundly modified the secretome content of MSCs, impairing the above-described activity and promoting the release of inflammatory factors., Conclusion: We demonstrated that the content of MSC secretomes depends on tissue microenvironment and that pathological condition may profoundly alter its composition. Video abstract.

Making a smooth transition to the K-12 (kindergarten through Grade 12) classroom context sets the stage for academic success throughout the life course. Parents' early education-related behaviors are linked with children's adjustment, yet less is known about how parental school readiness beliefs motivate parenting practices at this educational transition. We investigated the associations between parental school readiness beliefs (general and child-specific) following the transition to kindergarten and parents' involvement the following year. Using data from the Early Childhood Longitudinal Study-Kindergarten 2011 cohort ( N = 9,790), general school readiness beliefs and child-specific academic and behavioral competency beliefs were associated with school-based involvement in first grade. Kindergarten parents who held higher child-specific academic competency beliefs also reported less homework involvement and had greater teacher-reported classroom-based involvement in first grade. Family poverty status differences did not emerge. Findings can inform efforts to increase parental involvement by elucidating the ways in which parents' beliefs about their children motivate involvement strategies.

Autophagy is a highly conserved degradative process that conveys dysfunctional proteins, lipids, and organelles to lysosomes for degradation. The post-mitotic nature, complex and highly polarized morphology, and high degree of specialization of neurons make an efficient autophagy essential for their homeostasis and survival. Dysfunctional autophagy occurs in aging and neurodegenerative diseases, and autophagy at synaptic sites seems to play a crucial role in neurodegeneration. Moreover, a role of autophagy is emerging for neural development, synaptogenesis, and the establishment of a correct connectivity. Thus, it is not surprising that defective autophagy has been demonstrated in a spectrum of neurodevelopmental disorders, often associated with early-onset epilepsy. Here, we discuss the multiple roles of autophagy in neurons and the recent experimental evidence linking neurodevelopmental disorders with epilepsy to genes coding for autophagic/lysosomal system-related proteins and envisage possible pathophysiological mechanisms ranging from synaptic dysfunction to neuronal death., (Copyright © 2020 Fassio, Falace, Esposito, Aprile, Guerrini and Benfenati.)

The current study investigated early adolescents' experiences of friend-related stress across middle school and its developmental consequences following the transition to high school. Using a sample of approximately 1,000 middle school students, four unique friend-related stress trajectories were observed across middle school: consistently low friend-related stress (57% of the sample), consistently high friend-related stress (7%), moderate and increasing friend-related stress (22%), and moderate but decreasing friend-related stress (14%). Groups characterized by higher levels of friend-related stress across middle school were linked to subsequent poorer socioemotional well-being, lower academic engagement, and greater involvement in and expectancies around risky behaviors following the transition to high school. Increased friend-related stress across the high school transition was also linked to poorer outcomes, even after taking into account earlier stress trajectories. Gender differences highlighted the particular struggles girls experience both in friend stress and in the links between friend stress and subsequent well-being., Competing Interests: Declaration of Conflicting Interests The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: The ubiquitous diffusion of radiofrequency (RF) radiation across human living environments has attracted the attention of scientists. Though the adverse health effects of RF exposure remain debatable, it has been reported that the interaction of such radiation with biological macromolecular structures can be deleterious for stem cells, inducing impairment of their main functions involving self-renewal and differentiation., Purpose: The purpose of this study was to determine whether exposure to RF of 169 megahertz (MHz) that is part of very high radiofrequency (VHF) range 30-300 MHz, could cause damage to stem cells by inducing senescence and loss of regenerative and DNA repair capacity., Methods: The study was conducted on mesenchymal stromal cells (MSCs) containing a subpopulation of stem cells. The MSCs were exposed to RFs of 169 MHz administered via an open meter 2G "Smart Meter" for different durations of time., Result: We did not observe modifications in MSC biology as a result of the RF exposure conducted in our experiments., Conclusion: We concluded that MSCs are insensitive to RF radiation exposure at 169 MHz for various time intervals, including longer durations., Competing Interests: Massimo Briccola, Paolo Giubbini, and Raffaella Marchesani were employed by e-distribuzione SpA at the time of the study. The authors report no other conflicts of interest in this work., (© 2019 Alessio et al.)

Ohtahara syndrome, early infantile epileptic encephalopathy with a suppression burst EEG pattern, is an aetiologically heterogeneous condition starting in the first weeks or months of life with intractable seizures and profound developmental disability. Using whole exome sequencing, we identified biallelic DMXL2 mutations in three sibling pairs with Ohtahara syndrome, belonging to three unrelated families. Siblings in Family 1 were compound heterozygous for the c.5135C>T (p.Ala1712Val) missense substitution and the c.4478C>G (p.Ser1493*) nonsense substitution; in Family 2 were homozygous for the c.4478C>A (p.Ser1493*) nonsense substitution and in Family 3 were homozygous for the c.7518-1G>A (p.Trp2507Argfs*4) substitution. The severe developmental and epileptic encephalopathy manifested from the first day of life and was associated with deafness, mild peripheral polyneuropathy and dysmorphic features. Early brain MRI investigations in the first months of life revealed thin corpus callosum with brain hypomyelination in all. Follow-up MRI scans in three patients revealed progressive moderate brain shrinkage with leukoencephalopathy. Five patients died within the first 9 years of life and none achieved developmental, communicative or motor skills following birth. These clinical findings are consistent with a developmental brain disorder that begins in the prenatal brain, prevents neural connections from reaching the expected stages at birth, and follows a progressive course. DMXL2 is highly expressed in the brain and at synaptic terminals, regulates v-ATPase assembly and activity and participates in intracellular signalling pathways; however, its functional role is far from complete elucidation. Expression analysis in patient-derived skin fibroblasts demonstrated absence of the DMXL2 protein, revealing a loss of function phenotype. Patients' fibroblasts also exhibited an increased LysoTracker® signal associated with decreased endolysosomal markers and degradative processes. Defective endolysosomal homeostasis was accompanied by impaired autophagy, revealed by lower LC3II signal, accumulation of polyubiquitinated proteins, and autophagy receptor p62, with morphological alterations of the autolysosomal structures on electron microscopy. Altered lysosomal homeostasis and defective autophagy were recapitulated in Dmxl2-silenced mouse hippocampal neurons, which exhibited impaired neurite elongation and synaptic loss. Impaired lysosomal function and autophagy caused by biallelic DMXL2 mutations affect neuronal development and synapse formation and result in Ohtahara syndrome with profound developmental impairment and reduced life expectancy., (© The Author(s) (2019). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Mutations in TBC1D24 are described in patients with a spectrum of neurological diseases, including mild and severe epilepsies and complex syndromic phenotypes such as Deafness, Onycodystrophy, Osteodystrophy, Mental Retardation and Seizure (DOORS) syndrome. The product of TBC1D24 is a multifunctional protein involved in neuronal development, regulation of synaptic vesicle trafficking, and protection from oxidative stress. Although pathogenic mutations in TBC1D24 span the entire coding sequence, no clear genotype/phenotype correlations have emerged. However most patients bearing predicted loss of function mutations exhibit a severe neurodevelopmental disorder. Aim of the study is to investigate the impact of TBC1D24 knockdown during the first stages of neuronal differentiation when axonal specification and outgrowth take place. In rat cortical primary neurons silenced for TBC1D24, we found defects in axonal specification, the maturation of axonal initial segment and action potential firing. The axonal phenotype was accompanied by an impairment of endocytosis at the growth cone and an altered activation of the TBC1D24 molecular partner ADP ribosylation factor 6. Accordingly, acute knockdown of TBC1D24 in cerebrocortical neurons in vivo analogously impairs callosal projections. The axonal defect was also investigated in human induced pluripotent stem cell-derived neurons from patients carrying TBC1D24 mutations. Reprogrammed neurons from a patient with severe developmental encephalopathy show significant axon formation defect that were absent from reprogrammed neurons of a patient with mild early onset epilepsy. Our data reveal that alterations of membrane trafficking at the growth cone induced by TBC1D24 loss of function cause axonal and excitability defects. The axonal phenotype correlates with the disease severity and highlight an important role for TBC1D24 in connectivity during brain development.

Senescent cells secrete inflammatory cytokines, proteases, and other factors, which are indicated as senescence-associated secretory phenotype (SASP). There are contrasting studies on the role of the SASP in cancer. Studies suggested that cancer cells may misuse the senescent secretome for their growth. Other investigations evidenced that the SASP may induce cancer growth arrest, senescence, or apoptosis. These conflicting data can be reconciled considering that cancer cells can coax senescent cells to secrete factors for their survival, thus abrogating the SASP's anti-cancer effect. Cancer stage may also have an impact on the capacity of the SASP to block tumor proliferation and promote senescence. Indeed, senescence is associated with a permanent cell cycle arrest, which needs functional cell cycle checkpoints. We evaluated the SASP effect on the in vitro biological properties of PNT2 and PC3 cells, which are immortalized prostate cells and metastatic prostatic cancer cells, respectively. We evidenced that SASPs, coming either from mesenchymal stromal cells treated with H 2 0 2 or with low X-ray doses, induced senescence of immortalized cells but not of cancer cells. Hence, the SASP released by acute senescent cells should be considered as an effective weapon against pre-tumorigenesis events rather than an anti-cancer mechanism acting on malignant cells.

Mutations in the Tre2/Bub2/Cdc16 (TBC)1 domain family member 24 (TBC1D24) gene are associated with a range of inherited neurological disorders, from drug-refractory lethal epileptic encephalopathy and DOORS syndrome (deafness, onychodystrophy, osteodystrophy, mental retardation, seizures) to non-syndromic hearing loss. TBC1D24 has been implicated in neuronal transmission and maturation, although the molecular function of the gene and the cause of the apparently complex disease spectrum remain unclear. Importantly, heterozygous TBC1D24 mutation carriers have also been reported with seizures, suggesting that haploinsufficiency for TBC1D24 is significant clinically. Here we have systematically investigated an allelic series of disease-associated mutations in neurons alongside a new mouse model to investigate the consequences of TBC1D24 haploinsufficiency to mammalian neurodevelopment and synaptic physiology. The cellular studies reveal that disease-causing mutations that disrupt either of the conserved protein domains in TBC1D24 are implicated in neuronal development and survival and are likely acting as loss-of-function alleles. We then further investigated TBC1D24 haploinsufficiency in vivo and demonstrate that TBC1D24 is also crucial for normal presynaptic function: genetic disruption of Tbc1d24 expression in the mouse leads to an impairment of endocytosis and an enlarged endosomal compartment in neurons with a decrease in spontaneous neurotransmission. These data reveal the essential role for TBC1D24 at the mammalian synapse and help to define common synaptic mechanisms that could underlie the varied effects of TBC1D24 mutations in neurological disease.

The human chr15q11-q13 imprinted cluster is linked to several disorders, including Prader-Willi (PWS) and Angelman (AS) syndromes. Recently, disease modeling approaches based on induced pluripotent stem cells (iPSCs) have been used to study these syndromes. A concern regarding the use of these cells for imprinted disease modeling is the numerous imprinting defects found in many iPSCs. Here, by reprogramming skin fibroblasts from a control and AS individuals, we generated several iPSC lines and addressed the stability of imprinting status across the PWS/AS domain. We focused on three important regulatory DNA elements which are all differentially methylated regions (DMRs), methylated on the maternal allele: the PWS imprinting center (PWS-IC), which is a germline DMR and the somatic NDN and MKRN3 DMRs, hierarchically controlled by PWS-IC. Normal PWS-IC methylation pattern was maintained in most iPSC lines; however, loss of maternal methylation in one out of five control iPSC lines resulted in a monoallelic to biallelic switch for many imprinted genes in this domain. Surprisingly, MKRN3 DMR was found aberrantly hypermethylated in all control and AS iPSCs, regardless of the methylation status of the PWS-IC master regulator. This suggests a loss of hierarchical control of imprinting at PWS/AS region. We confirmed these results in established iPSC lines derived using different reprogramming procedures. Overall, we show that hierarchy of imprinting control in donor cells might not apply to iPSCs, accounting for their spectrum of imprinting alterations. Such differences in imprinting regulation should be taken into consideration for the use of iPSCs in disease modeling.

Synaptic transmission is critically dependent on synaptic vesicle (SV) recycling. Although the precise mechanisms of SV retrieval are still debated, it is widely accepted that a fundamental role is played by clathrin-mediated endocytosis, a form of endocytosis that capitalizes on the clathrin/adaptor protein complex 2 (AP2) coat and several accessory factors. Here, we show that the previously uncharacterized protein KIAA1107, predicted by bioinformatics analysis to be involved in the SV cycle, is an AP2-interacting clathrin-endocytosis protein (APache). We found that APache is highly enriched in the CNS and is associated with clathrin-coated vesicles via interaction with AP2. APache-silenced neurons exhibit a severe impairment of maturation at early developmental stages, reduced SV density, enlarged endosome-like structures, and defects in synaptic transmission, consistent with an impaired clathrin/AP2-mediated SV recycling. Our data implicate APache as an actor in the complex regulation of SV trafficking, neuronal development, and synaptic plasticity., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

The transition to high school is disruptive for many adolescents, yet little is known about the supportive relational processes that might attenuate the challenges students face as they move from middle to high school, particularly for students from more diverse backgrounds. Identifying potential buffers that protect youth across this critical educational transition is important for informing more effective support services for youth. In this study, we investigated how personal characteristics (gender, nativity, parent education level) and changes in support from family, friends, and school influenced changes in socioemotional adjustment and academic outcomes across the transition from middle to high school. The data were drawn from 252 students (50% females, 85% Latina/o). The results revealed declines in students' grades and increases in depressive symptoms and feelings of loneliness across the high school transition, with key variation by student nativity and gender. Additionally, stable/increasing friend support and school belonging were both linked to less socioemotional disruptions as students moved from middle to high school. Increasing/stable school belonging was also linked to increases in school engagement across the high school transition. These findings suggest that when high school transitions disrupt supportive relationships with important others in adolescents' lives, adolescents' socioemotional well-being and, to a lesser extent, their academic engagement are also compromised. Thus, in designing transition support activities, particularly for schools serving more low-income and race/ethnic minority youth, such efforts should strive to acclimate new high school students by providing inclusive, caring environments and positive connections with educators and peers.

Compared to white girls, sexual maturation is accelerated in African American girls as measured by indicators of pubertal development, including age at first menses. Increasing epidemiological evidence suggests that the timing of pubertal development may have strong implications for cardio-metabolic health in adolescence and adulthood. In fact, younger menarcheal age has been related prospectively to poorer cardiovascular risk factor profiles, a worsening of these profiles over time, and an increase in risk for cardiovascular events, including non-fatal incident cardiovascular disease and cardiovascular-specific and all-cause mortality. Yet, because this literature has been limited almost exclusively to white girls/women, whether this same association is present among African American girls/women has not been clarified. In the current narrative review, the well-established vulnerability of African American girls to experience earlier pubertal onset is discussed as are findings from literatures examining the health outcomes of earlier pubertal timing and its antecedents, including early life adversity exposures often experienced disproportionately in African American girls. Gaps in these literatures are highlighted especially with respect to the paucity of research among minority girls/women, and a conceptual framework is posited suggesting disparities in pubertal timing between African American and white girls may partially contribute to well-established disparities in adulthood risk for cardio-metabolic disease between African American and white women. Future research in these areas may point to novel areas for intervention in preventing or lessening the heightened cardio-metabolic risk among African American women, an important public health objective.

TBC1D24-related disorders include a wide phenotypic ranging from mild to lethal seizure disorders, non-syndromic deafness, and composite syndromes such as DOORS (deafness, onychodystrophy, osteodystrophy, mental retardation, and seizures). The TBC1D24 gene has a role in cerebral cortex development and in presynaptic neurotransmission. Here, we present a familial case of a lethal early-onset epileptic encephalopathy, associated with two novel compound heterozygous missense variants on the TBC1D24 gene, which were detected by exome sequencing. The detailed clinical data of the three siblings is summarized in order to support the variability of the phenotype, severity, and progression of this disorder among these family members. Functional studies demonstrated that the identified novel missense mutations result in a loss of expression of the protein, suggesting a correlation between residual expression, and the disease severity. This indicates that protein expression analysis is important for interpreting genetic results when novel variants are found, as well as for complementing clinical assessment by predicting the functional impact. Further analysis is necessary to delineate the clinical presentation of individuals with TBC1D24 pathogenic variants, as well as to develop markers for diagnosis, prognosis, and potential targeted treatments. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Unlabelled: Electroencephalography (EEG) is useful to objectively diagnose/grade hepatic encephalopathy (HE) across its spectrum of severity. However, it requires expensive equipment, and hepatogastroenterologists are generally unfamiliar with its acquisition/interpretation. Recent technological advances have led to the development of low-cost, user-friendly EEG systems, allowing EEG acquisition also in settings with limited neurophysiological experience. The aim of this study was to assess the relationship between EEG parameters obtained from a standard-EEG system and from a commercial, low-cost wireless headset (light-EEG) in patients with cirrhosis and varying degrees of HE. Seventy-two patients (58 males, 61 ± 9 years) underwent clinical evaluation, the Psychometric Hepatic Encephalopathy Score (PHES), and EEG recording with both systems. Automated EEG parameters were calculated on two derivations. Strong correlations were observed between automated parameters obtained from the two EEG systems. Bland and Altman analysis indicated that the two systems provided comparable automated parameters, and agreement between classifications (normal versus abnormal EEG) based on standard-EEG and light-EEG was good (0.6 < κ < 0.8). Automated parameters such as the mean dominant frequency obtained from the light-EEG correlated significantly with the Model for End-Stage Liver Disease score (r = -0.39, P < 0.05), fasting venous ammonia levels (r = -0.41, P < 0.01), and PHES (r = -0.49, P < 0.001). Finally, significant differences in light-EEG parameters were observed in patients with varying degrees of HE., Conclusion: Reliable EEG parameters for HE diagnosing/grading can be obtained from a cheap, commercial, wireless headset; this may lead to more widespread use of this patient-independent tool both in routine liver practice and in the research setting. (, (© 2016 by the American Association for the Study of Liver Diseases.)

The negative-ion accelerator for the MITICA neutral beam injector has been designed and optimized in order to reduce the thermo-mechanical stresses in all components below limits compatible with the required fatigue life. However, deviation from the expected beam performances can be caused by "off-normal" operating conditions of the accelerator. The purpose of the present work is to identify and analyse all the "off-normal" operating conditions, which could possibly become critical in terms of thermo-mechanical stresses or of degradation of the optical performances of the beam.

Purpose: The goal of this study was test expectations derived from sociological and developmental perspectives that the association between phonics instruction in kindergarten classrooms and reading achievement during the first year of school in the low-income population would depend on whether children had previously attended preschool as well as the socioeconomic composition of their elementary schools., Methodological Approach: Autoregressive modeling was applied to nationally representative data from 7,710 children from low-income families in the Early Childhood Longitudinal Study-Kindergarten Cohort, with a series of sensitivity tests to improve causal inference and explore the robustness of results., Findings: The association between phonics instruction and achievement was strongest among children from low-income families who had not attended preschool and then enrolled in socioeconomically disadvantaged elementary schools and among children from low-income families who had attended preschool and then enrolled in socioeconomically advantaged elementary schools., Research and Practical Implications: Insight into educational inequality can be gained by situating developing children within their proximate ecologies and institutional settings, especially looking to the match between children and their contexts. These findings are relevant to policy discussions of early education, instructional practices, and desegregation.

The present study describes the generation and the characterization of a stable cell line of neural stem cells derived from embryonic mouse hypothalamus. These cells (AC1) grow as an adherent culture in defined serum-free medium and express typical markers of neurogenic radial glia and of hypothalamic precursors. After prolonged expansion, AC1 cells may be efficiently induced to differentiate into neurons and astroglial cells in vitro and start to express some hormonal neuropeptides, like TRH, CRH, and POMC. Based on the capabilities of AC1 cells to be stably expanded and to develop neuroendocrine lineages in vitro, these cells might represent a novel tool to elucidate the mechanisms involved in the development of the hypothalamus and in the specific differentiation of neuroendocrine neurons.

The relationship between hepatic encephalopathy (HE) and the sleep-wake disturbances exhibited by patients with cirrhosis remains debated. The aim of this study was to examine the usefulness of sleep-wake interview within the context of HE assessment. One-hundred-and-six cirrhotic patients were asked three yes/no questions investigating the presence of difficulty falling asleep, night awakenings and daytime sleepiness. All underwent formal HE assessment, quantitative electroencephalography and standardised psychometry. Fifty-eight were monitored for 8 ± 6 months in relation to the occurrence of HE. Patients complaining of daytime sleepiness (n = 75, 71 %) had slower EEGs than those who did not report it (relative alpha power: 37 ± 19 vs. 48 ± 17 %, p < 0.05). In addition, daytime sleepiness was associated with the presence of portal-systemic shunt (79 vs. 57 %, p < 0.05) and HE history (72 vs. 45 %, p < 0.05). Finally, the absence of excessive daytime sleepiness had a Negative Predictive Value of 92 % (64-100) in relation to the development of HE during the follow-up period. These data support the appropriateness of adding a yes/no question on the presence of excessive daytime sleepiness to routine assessment of patients with cirrhosis, to help identify those who do not need further, formal HE screening.

Aim: The aim of this randomized controlled trial was to assess whether action observation treatment may improve upper limb motor functions in children with cerebral palsy (CP)., Method: All children with CP admitted to our unit for rehabilitation from May 2009 to May 2010 were eligible. Inclusion criteria were age between 6 years and 11 years, an IQ of at least 70, and no major visual and/or auditory deficits. Fifteen children were enrolled and randomly assigned to either a case group (n=8; four males, four females; median age 7 y 6 mo) or control group (n=7; five males, two females; median age 8 y). Six participants had left-sided hemiplegia, six right-sided hemiplegia, and three had tetraplegia; 10 were independent walkers. Those in the case group were asked to observe video clips showing daily age-appropriate actions, and afterwards to imitate them. Participants in the control group were asked to observe video clips with no motor content and afterwards to execute the same actions as cases. The primary outcome measure was the Melbourne Assessment Scale. Children were scored twice at baseline (2 wks apart), and at the end of treatment, by a physician blind to group assignment., Results:   At baseline groups did not differ on functional evaluation. After treatment, the functional score gain (Δ) was significantly different in the case and control groups (p=0.026)., Interpretation: The present results support the notion that action observation treatment can be an effective part of the rehabilitation programme in children with CP., (© The Authors. Developmental Medicine & Child Neurology © 2012 Mac Keith Press.)

This longitudinal study examined Latino adolescents' feelings of loneliness and the repercussions of loneliness for later educational success. Participants were 640 Latino students (56% girls, 62% Mexican/Mexican-American) who reported on loneliness across the first 2 years of high school. Growth mixture modeling identified three distinct loneliness trajectory classes for the Latino adolescents--consistently low, chronically high, and low but increasing. Language brokering, language use, and school mobility emerged as predictors of class membership. Increasingly and chronically lonely youth experienced academic difficulty, both in terms of academic progress and exit exam success, but support from friends served as a buffer of the negative relationship between loneliness and academic success. This study highlights the pernicious effects of loneliness and suggests promoting prosocial friendship support as a means of facilitating more positive academic outcomes for Latino youth.

This study examined the role of adolescents' and mothers' self-reports of English and heritage language proficiency in youth's academic and emotional adjustment among 444 Chinese American families. Adolescents who were proficient in English tended to exhibit higher reading achievement scores, math achievement scores, and overall GPA. Mothers who were English proficient tended to have children with higher academic achievement and fewer depressive symptoms. Results also indicated that adolescents' heritage language maintenance was associated with positive adjustment, particularly amongst foreign-born youth and for youth whose parents were highly proficient in the heritage language. Mother-adolescent match in heritage language proficiency was related to higher math achievement scores and overall GPA. Additionally, higher heritage language proficiency was associated with fewer depressive symptoms for foreign-born but not U.S.-born youth. Overall, the findings suggest that proficiency in both the English and heritage language may confer advantages to Chinese American youth.

Researchers, service providers, and policymakers must uncover and better understand the issues facing youths in Asian gangs in order to most effectively intervene with appropriate policies and programs. The present investigation sampled young male Filipino gang members in Hawai'i. Thematic analyses of the focus group data challenge the commonly held view of racial harmony in Hawai'i. It appears that racial and social discrimination from peers and authority figures propel Filipino boys to seek out gang membership as a way to protect themselves from being targets of oppression.

The sociometric and theoretical models traditionally associated with the determinants of unionisation and based on international comparisons, generally use linear analysis of a restricted number of complex variables. The effectiveness of such models often have an unsatisfactory result. In this study a different theoretical and methodological approach has been used. It is based on a integrated scheme made by transposing the principles of the "Squashing Theory" (Buscema, 1994) into the social field, by the connectionist paradigm, and by the Artificial Neural Networks (ANN). This has permitted the carrying out of a larger data base (49 variables, 12 Nations, 12 Years maximum time interval), the adjusting of a model that has an elevated explanatory capacity (in terms of open variance) an ulterior capacity of generalization (in terms of a one-year span) and of metageneralization (in terms of generalization to nonprocessed nations). What emerges is surely the need for more and greater in-depth study of socializing facts which follow such aspects.