Effects of victimization and perpetration in observing bullying scenes: an eye-tracker study

Introduction: Previous research showed that bullying experiences are associated with different ways of interpreting and behaving in bullying dynamics. However, it remains uncertain whether these distinctions can already be present during the first step of information processing: the allocation of attention. Aims: The study explored attentional patterns of Italian students with different bullying experiences in daily life while observing different roles represented through bullying vignettes. Methods: Participants (72 students, Mage= 11.18) were categorized as victims, bully-victims, or not involved based on their scores on a self-report questionnaire. They observed 9 bullying vignettes on which different portraits were presented (bully, victim, pro-bully, defender, bystander) while the eye-tracker registered attentional indexes (fixation, visit and duration). Results: Kruskal- Wallis and pairwise comparisons revealed a significant effect for the portraits of the bully and the pro-bully as bully-victims exhibited greater fixations and visits than victims, while students not involved showed no significant differences with the other groups. Conclusion: Our research reveals that bully-victims focused more on threatening cues while victims diverged their gaze from them, confirming that the experience of bullying influences how they explore aggressive situations. Learning how involved students direct their attention helps us understand different responses, leading to powerful interventions

Congenital Adrenal Hyperplasias Presenting in the Newborn and Young Infa Front Pediatr. 2020 Dec 22;8:593315. doi: 10.3389/fped.2020.593315. eCollection 2020. PMID: 33415088

Congenital adrenal hyperplasia includes autosomal recessive conditions that affect the adrenal cortex steroidogenic enzymes (cholesterol side-chain cleavage enzyme; 3b-hydroxysteroid dehydrogenase; 17a-hydroxylase/17,20 lyase; P450 oxidoreductase; 21-hydroxylase; and 11b-hydroxylase) and proteins (steroidogenic acute regulatory protein). These are located within the three major pathways of the steroidogenic apparatus involved in the production of mineralocorticoids, glucocorticoids, and androgens. Many countries have introduced newborn screening program (NSP) based on 17-OH-progesterone (17-OHP) immunoassays on dried blood spots, which enable faster diagnosis and treatment of the most severe forms of 21-hydroxylase deficiency (21-OHD). However, in several others, the use of this diagnostic tool has not yet been implemented and clinical diagnosis remains challenging, especially for males. Furthermore, less severe classic forms of 21-OHD and other rarer types of CAHs are not identified by NSP. The aim of this mini review is to highlight both the main clinical characteristics and therapeutic options of these conditions, which may be useful for a differential diagnosis in the neonatal period, while contributing to the biochemical evolution taking place in the steroidogenic field. Currently, chromatographic techniques coupled with tandem mass spectrometry are gaining attention due to an increase in the reliability of the test results of NPS for detecting 21-OHD. Furthermore, the possibility of identifying CAH patients that are not affected by 21-OHD but presenting elevated levels of 17-OHP by NSP and the opportunity to include the recently investigated 11-oxygenated androgens in the steroid profiles are promising tools for a more precise diagnosis and monitoring of some of these conditions

A sequence variation in 3\u2019UTR of CYP21A2 gene correlates with a mild form of Congenital Adrenal Hyperplasia

Congenital adrenal hyperplasia is mainly caused by the deficiency of the 21-hydroxylase enzyme coded by the CYP21A2 gene. However, some alleles inthe nonclassical form (NC-CAH) remain without identified mutations, suggesting the involvement of regulatory regions. Our objective was to study an allele carrying the variant *13 G>A in the 3\u2019UTR of the CYP21A2 gene identified in some patients with NC-CAH in order to verify the possible implication of this variation with the phenotype observed. From all the subjects in whom the CYP21A2 gene was analysed, 14 patients and 7 relatives heterozygous or homozygous for the *13 G>A substitution in 3\u2019UTR were selected. Sequencing of DNA, genotyping, multiplex ligation-dependent probe amplification (MLPA), in-vitro studies and bioinformatic analysis were performed.The haplotype of the *13 G>A allele resulted identical in all the subjects and no other concomitant mutations were found in the region extending from 3 Kb upstream the starting codon to the polyadenilation signal. MLPA analysis showed the monomodular structure of the allele composed by one C4A gene and one CYP21A2 gene without a second module with the CYP21A1P pseudogene. Bioinformatic analysis predicted a consistent alteration of the RNA folding due to this sequence variation and no miRNA target sequence in this region. In vitro studies confirmed a decreased expression level for the mRNA carrying the *13 G>A variation . In conclusion, this substitution in the 3\u2019UTR of the gene seems to be implicated in the NC-CAH phenotype suggesting the importance of analysing also the untranslated regions

TRANSGLUTAMINASE-CATALYZED POLYMERIZATION OF TROPONIN IN-VITRO

In the presence of calcium ions, tissue transglutaminase catalyzes the polymerization of skeletal muscle troponin to high molecular weight insoluble aggregate. The specific action of transglutaminase is proved by the isolation of glutamyl-spermidine isopeptide derivatives. The process involves mainly the troponin T subunit (TnT), with formation of dimers and trimers of TnT, which were reactive with specific antibodies by immunoblotting. Furthermore when incubation is carried out in the presence of radioactive polyamines, the label is incorporated selectively into TnT subunit

Overexpression of the stress-protein Grp94 reduces cardiomyocyte necrosis due to calcium overload and simulated ischemia

Increase in free intracellular calcium [Ca2+](i) plays a crucial role in cardiomyocyte ischemic injury. Here we demonstrate that overexpression of the sarcoplasmic-reticulum stress-protein Grp94 reduces myocyte necrosis due to calcium overload or simulated ischemia. Selective three-to eightfold Grp94 increase, with no change in Grp78 or calreticulin amount, was achieved by stable transfection of skeletal C2C12 and cardiac H9c2 muscle cells. After exposure to the calcium ionophore A23187, LDH release from five different Grp94-overexpressing clones of either C2C12 and H9c2 origin was significantly lower than that of control ones and [Ca2+](i) increase was significantly delayed. The number of necrotic cells, evaluated by propidium iodide uptake, was reduced when cells from the Grp94-overexpressing H9c2 clone were exposed to conditions simulating ischemia. Experiments performed in neonatal rat cardiomyocytes cotransfected with grp94 and the green fluorescent protein (GFP) cDNAs validated the protective effect of Grp94 overexpression. A lower percentage of propidium-iodide positive/GFP-fluorescent myocytes co-expressing exogenous Grp94, with respect to myocytes expressing GFP alone, was observed after exposure to either A23187 (6.6% vs. 14.0%, respectively) or simulated ischemia (8.5% vs. 17.7%, respectively). In conclusion, the selective increase in Grp94 protects cardiomyocytes from both ischemia and calcium overload counteracting [Ca2+](i) elevations

Improving the diagnosis of 11β-hydroxylase deficiency using home-made MLPA probes: identification of a novel chimeric CYP11B2/CYP11B1 gene in a Sicilian patient

Purpose: 11β-Hydroxylase deficiency (11OHD) represents the second most common cause of congenital adrenal hyperplasia. It is caused by mutations in the CYP11B1 gene localized about 40 kb from the CYP11B2 gene with which it shares a homology of 95 %. The asymmetric recombination of these two genes is involved both in 11OHD and in glucocorticoid-remediable aldosteronism (GRA). Our objective was to set up an easy and rapid method to detect these hybrid genes and other kinds of deletions, to improve the molecular diagnosis of 11OHD. Methods: A set of 8 specific probes for both the CYP11B1 and the CYP11B2 genes to be used for multiplex ligation-dependent probe amplification (MLPA) analysis was designed to detect rearrangements of these genes. Results: The method developed was tested on 15 healthy controls and was proved to be specific and reliable; it led us to identify a novel chimeric CYP11B2/CYP11B1 gene in one patient that carried the known A306V mutation on the other allele. Specific amplification and sequencing of the hybrid gene confirmed the breakpoint localization in the second intron. Conclusions: The MLPA kit developed enables the detection of deletions, duplications or chimeric genes and represents an optimal supplement to DNA sequence analysis in patients with 11OHD. In addition, it can also be used to show the presence of the opposite chimaera associated with GRA