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Here we present a novel method "Genomic inverse PCR for exploration of ligated breakpoints" (GIPFEL) that allows the sensitive detection of recurrent chromosomal translocations. This technique utilizes limited amounts of DNA as starting material and relies on PCR based quantification of unique DNA sequences that are created by circular ligation of restricted genomic DNA from translocation bearing cells. Because the complete potential breakpoint region is interrogated, a prior knowledge of the individual, specific interchromosomal fusion site is not required. We validated GIPFEL for the five most common gene fusions associated with childhood leukemia (MLL-AF4, MLL-AF9, MLL-ENL, ETV6-RUNX1, and TCF3-PBX1). A workflow of restriction digest, purification, ligation, removal of linear fragments and precipitation enriching for circular DNA was developed. GIPFEL allowed detection of translocation specific signature sequences down to a 10-4 dilution which is close to the theoretical limit. In a blinded proof-of-principle study utilizing DNA from cell lines and 144 children with B-precursor-ALL associated translocations this method was 100% specific with no false positive results. Sensitivity was 83%, 65%, and 24% for t(4;11), t(9;11) and t(11;19) respectively. Translocation t(12;21) was correctly detected in 64% and t(1;19) in 39% of the cases. In contrast to other methods, the characteristics of GIPFEL make it particularly attractive for prospective studies.

Current diagnostic approaches that characterize T-cell deficiency by analyzing diversity of T-cell receptor sequences effectuate limited informational gain about the actual restrictiveness. For deeper insight into T-cell receptor repertoires we developed next-generation-sequencing-spectratyping, which employs high coverage Roche/454 sequencing of T-cell receptor (β)-chain amplicons. For automated analysis of high-throughput-sequencing data, we developed a freely available software, the TCR profiler. Gene usage, length, encoded amino acid sequence and sequence diversity of the complementarity determining region 3 were determined and comprehensively integrated into a novel complexity score. Repertoires of CD8+ T cells from children with idiopathic or hepatitis-induced very severe aplastic anemia (n=7), children two months after bone marrow transplantation (n=7) and healthy controls (children n=5, adults n=5) were analyzed. Complexity scores clearly distinguished between healthy and diseased, and even between different immune deficiency states. The repertoire of aplastic anemia patients was dominated by public (i.e. present in more than one person) T-cell receptor clonotypes, whereas only 0.2% or 1.9% were public in normal children and adults, respectively. The CDR3 sequence ASSGVGFSGANVLT was highly prevalent in 3 cases of hepatitis-induced anemia (15–32% of all sequences), but was only low expressed in idiopathic aplastic anemia (2–5%, n=4) or healthy controls (

Abstract Aim of the present study is to evaluate the relationship between genetic and phenotypic data in a series of patients affected by grade I and II of foveal hypoplasia with stable fixation and good visual acuity using multimodal imaging techniques. All patients underwent complete clinical and instrumental assessment including structural Optical Coherence Tomography (OCT), OCT Angiography and Adaptive Optics (AO) imaging. Central macular thickness (CMT), inner nuclear layer (INL), vessel density in superficial capillary plexus were the main variables evaluated with OCT technology. Cone density, cone spacing, cone regularity, cone dispersion and angular density were the parameters evaluated with AO. Genetic evaluation and trio exome sequencing were performed in all affected individuals. Eight patients (3 males and 5 females) with a mean age of 12.62 years (range 8–18) were enrolled. The mean best corrected visual acuity (BCVA) was 0.18 ± 0.13 logMAR, mean CMT was 291.9 ± 16.6 µm and INL was 26.2 ± 4.6 µm. The absence of a foveal avascular zone (FAZ) was documented by examination of OCT-A in seven patients in the superficial capillary plexus. However, there was a partial FAZ in the deep plexus in patients P5 and P8. Of note, all the patients presented with major retinal vessels clearly crossing the foveal center. All individuals exhibited a grade I or II of foveal hypoplasia. In 5 patients molecular analyses showed an extremely mild form of albinism caused by compound heterozygosity of a TYR pathogenic variant and the hypomorphic p.[Ser192Tyr;Arg402Gln] haplotype. One patient had Waardenburg syndrome type 2A caused by a de novo variant in MITF. Two patients had inconclusive molecular analyses. All the patients displayed abnormalities on OCT-A. Photoreceptor count did not differ from normal subjects according to the current literature, but qualitative analysis of AO imaging showed distinctive features likely related to an abnormal pigment distribution in this subset of individuals. In patients with foveal hypoplasia, genetic and multimodal imaging data, including AO findings, can help understand the physiopathology of the foveal hypoplasia phenotype. This study confirms that cone density and visual function can both be preserved despite the absence of a pit.

Backgrounds: Perfectionism is a multifaceted personality disposition characterized by striving for perfection, pursuing exceedingly high standards, and making highly critical self-evaluations. This disposition can impact mental and behavioral health and influence learning behavior in students. Perfectionism can foster a mastery orientation, where students commit to challenges and maintain confidence despite failures, or lead to learned helplessness, where students give up in the face of failure, attributing it to a lack of ability. Perfectionistic pressures from the environment, such as those perceived from teachers, may influence students’ personal perfectionism and learning behavior, and, in turn, their academic achievement. Methods: A structural equation modelling approach with latent variables was used to investigate the sequential mediating role of personal perfectionism (standard, order, and discrepancy) and learned helplessness/mastery orientation in the relationship between students’ perceptions of teacher perfectionism and their school achievements. We hypothesize that a mismatch between teachers’ expectations or evaluations and students’ actual performance (Teacher Discrepancy) may lead to students experiencing a gap between their self-perception and their actual abilities (Personal Discrepancy). This misalignment could result in feelings of helplessness and reduced achievement. On the other hand, if students perceive their teachers as having high standards and maintaining good order (adaptive perfectionism), this could encourage students to develop similar high standards and organizational skills (adaptive perfectionism), fostering a mastery-oriented mindset and ultimately enhancing their achievement. The study included 505 Italian high school students (Mage=16; SD =1.35). The instruments used were the Teacher Almost Perfect Scale, the Italian version of the Almost Perfect Scale-Revised, and the Learned Helplessness Questionnaire. Results: The SEM analysis revealed significant indirect effects. From Teacher Standards to school achievement via Personal Standards and mastery orientation (β = .12, p ≤ .01); from Teacher Discrepancy to School Achievement via Personal Standards and mastery orientation (β = -.04, p ≤ .01); from Teacher Discrepancy to School Achievement via Personal Discrepancy and mastery orientation (β = -.04, p ≤ .01). Conclusions: The findings highlight the roles of personal perfectionism (standards and discrepancy) and learning behaviors (learned helplessness/mastery orientation) in mediating the relationship between students’ perceptions of their teachers’ perfectionism and their academic achievement. This suggests that both the positive and negative dimensions of perceived teacher perfectionism can significantly impact students’ learning outcomes, mediated through their own perfectionistic tendencies. This study extends knowledge of factors that can impact students’ mental health, with important practical implications for prevent internalizing psychological disorders such as depression and anxiety.

A positive school climate promotes regular attendance through strong relationships, a sense of belonging, diversity promotion, clear rules, and safety, leading to improved relational and educational outcomes. This study explores how perceived academic and interpersonal competence mediate the relationship between school climate components and absenteeism, using Havik’s(2015) model. A cross-sectionalstudy of 539 Italian high school students employed structural equation modelling with latent variables to examine these mediations. Results indicated that perceptions of interpersonal competence mediated the relationship between Peer Connectedness ( =.16,p

Spectroscopic and photophysical characterization of the two new polyaminic derivatives possessing naphthalene as signaling units of the light induced molecular movements are investigated. It is shown that the central spacer unit, with different connections to the central benzene with the polyaminic chain, leads to small but significant differences in the photophysical behavior of the two compounds.

Abstract Background Central retinal artery occlusion (CRAO) is an ophthalmic emergency, and its etiology is generally ascribed to vessel occlusion by a thrombus or embolus, eventually due to a hypercoagulable state. CRAO occurrence is described even in the pediatric population, but its incidence is very rare. SARS-CoV-2 infection has a multitude of presentations, and almost any organ may be involved including the ocular district. Cases of CRAO in patients affected by COVID-19 are reported in the literature in the adult population, but not in the pediatric one. Case presentation We describe the case of a six-year-old otherwise healthy girl, who presented a sudden and complete bilateral vision loss after a one-day fever. All the clinical, ophthalmological, laboratory and instrumental investigations led to the diagnosis of a right CRAO and the suspicion of a contralateral posterior optic nerve affection. These manifestations could not be ascribed to any etiological condition apart from the documented ongoing mild SARS-CoV-2 infection. Treatment with anticoagulants and steroids was tried but the visual outcome was poor during the one-month hospitalization and at the last follow-up. Conclusions To the best of our knowledge, this is the first report of CRAO in the course of SARS-CoV-2 infection in the pediatric age. In our review of the literature, we found few cases of CRAO in adults with COVID-19; we highlighted differences in anamnestic, clinical, and interventional aspects and therefore we tried to summarize the state of the art on this topic to facilitate further studies. Even if rare, the prognosis of CRAO is poor and the thrombolytic treatment could be effective only if rapidly administered, so the disease suspicion should be high in a patient with sudden vision loss, also in pediatric age.

Background/Objectives: Burden Syndrome, also known as Caregiver Syndrome, particularly affects those who serve in the role of informal caregiver in the presence of family members with conditions. The ABCX dual model examines the impact on the caregiver of the diagnosis of autism spectrum disorder (ASD) on the family. This model considers the severity of the stressor (A), the additional demands of life stress (aA), the family’s internal resources (B), the family’s external resources (bB), the family’s assessment of the situation (C), coping strategies (cC), and outcome (X). The purpose of the present study is to investigate the relationships between resilience, guilt, and burden of care in caregivers of children with ASD. Methods: Various assessment instruments were used, including the “Caregiver Burden Inventory” to measure burden, the “Brief Resilience Scale” to assess resilience, the “Guilt Sensitivity Questionnaire” to examine guilt sensitivity, and the “DA.L.I.A.” to collect information on parent and child characteristics. A total of 80 parents/caregivers participated in the research, including 53 women (Age M = 41.72; SD = 7.8) and 27 men (Age M = 43.35; SD = 6.29). Results: The findings indicate that individuals’ resilience to stressful events correlates negatively with burden, a developmental subtype. However, guilt seems not to play a significant role in the overall perception of burden. In contrast, it was found that the use of informal supports is associated with higher levels of guilt and emotional burden, whereas the use of formal supports is correlated with higher emotional burden, but not higher perceptions of guilt. Conclusions: This study provides important information about the support needed by caregivers and suggests how to address emotional burdens to prevent burnout and support families with children with ASD.

Abstract Background Tumor hypoxia stimulates release of extracellular vesicles (EVs) that facilitate short- and long-range intercellular communication and metastatization. Albeit hypoxia and EVs release are known features of Neuroblastoma (NB), a metastasis-prone childhood malignancy of the sympathetic nervous system, whether hypoxic EVs can facilitate NB dissemination is unclear. Methods Here we isolated and characterized EVs from normoxic and hypoxic NB cell culture supernatants and performed microRNA (miRNA) cargo analysis to identify key mediators of EVs biological effects. We then validated if EVs promote pro-metastatic features both in vitro and in an in vivo zebrafish model. Results EVs from NB cells cultured at different oxygen tensions did not differ for type and abundance of surface markers nor for biophysical properties. However, EVs derived from hypoxic NB cells (hEVs) were more potent than their normoxic counterpart in inducing NB cells migration and colony formation. miR-210-3p was the most abundant miRNA in the cargo of hEVs; mechanistically, overexpression of miR-210-3p in normoxic EVs conferred them pro-metastatic features, whereas miR-210-3p silencing suppressed the metastatic ability of hypoxic EVs both in vitro and in vivo. Conclusion Our data identify a role for hypoxic EVs and their miR-210-3p cargo enrichment in the cellular and microenvironmental changes favoring NB dissemination.

Instagram is an image-sharing social networking service (SNS) particularly enjoyed by young adults. It excessive and problematic use impacts users’ body image (BI). Individuals with negative BI feel dissatisfied with their appearance and body, and perceive a discrepancy between their current and ideal image. Instagram, in contrast to other SNS, focused uniquely on image-based content. As such, it is likely that Instagram differs from other social media in term of mediating processes linking its use to BI, reflecting the need to understanding the relationship between maladaptive Instagram use and body dissatisfaction. The main purpose of this study is to investigate, through a self-determination theory (SDT) framework, the mediating role of basic psychological needs (BPN) satisfaction/frustration and needs-supportive/needs-thwarting interpersonal behaviors on the relationship between maladaptive Instagram use and body dissatisfaction. The results showed the role of BPN satisfaction/frustration (autonomy, competence and relatedness) in the relationship between maladaptive Instagram use and body dissatisfaction. The sample comprised 525 young adults from several Italian university. This study extends the knowledge of the factors that can be related to the BI of young adults, with important application implications.