Educator Perspectives on Mental Health Supports at the Primary Level.

During the early years of formal education, young students develop a number of formative academic, motor, behavioral, and socioemotional skills that lay the foundation for future learning. Since student mental health in the early grades predicts academic achievement in later grades, mental health interventions are essential at the primary school level. Not only are teachers expected to provide academic instruction, they are now involved in providing students with mental health services, despite a lack of training to do so. The current study sought to gather the perspectives of 38 primary-level educators to gain understanding about mental health knowledge, current approaches to mitigating mental health challenges, and barriers that prevent them from successfully addressing student mental health issues. Using thematic analysis, three themes developed: (1) Educators indicate supporting primary students' mental health is within their role; (2) Systems-level constraints prevent effective mental health supports; and (3) Staff desire increased mental health resources. Implications for educators and practice are discussed.

Importance, quality, and engagement: School mental health providers' perceptions regarding within-district transition care coordination practices.

Although care coordination (CC; i.e., the organization of care activities between professionals to facilitate appropriate service delivery; McDonald et al., 2007) has yet to be studied extensively within schools, preliminary research suggests coordinating school mental health supports can be beneficial (Francis et al., 2021) and that interprofessional and interagency collaboration is warranted to meet student needs (McClain et al., 2022). We examined the perceptions of school mental health providers (SMHPs) regarding importance, quality, and engagement with within-district transition CC practices within a multitiered system of support framework. Participants were 163 SMHPs who endorsed being involved in designing, providing, or implementing mental health services in a U.S. school district. The three scales used to measure engagement with CC practices were based on the Care Coordination Measures Atlas (McDonald et al., 2014) and were found to have promising preliminary psychometrics. Descriptive statistics indicated SMHPs endorsed CC as very important but perceived school and district personnel to view it as less important, reported their own quality of CC was slightly above that of their school and district, and regularly engaged in broad CC practices. Moreover, bivariate correlations indicated SMHP's personal views of CC importance were not associated with the quality of school and district CC, yet engagement in broad CC activities was associated with transition facilitation practices, and attitudes about CC were associated with engagement in broad CC activities. Implications of findings are discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Cys-X scanning for expansion of active-site residues and modulation of catalytic functions in a glutathione transferase.

We propose Cys-X scanning as a semisynthetic approach to engineer the functional properties of recombinant proteins. As in the case of Ala scanning, key residues in the primary structure are identified, and one of them is replaced by Cys via site-directed mutagenesis. The thiol of the residue introduced is subsequently modified by alternative chemical reagents to yield diverse Cys-X mutants of the protein. This chemical approach is orthogonal to Ala or Cys scanning and allows the expansion of the repertoire of amino acid side chains far beyond those present in natural proteins. In its present application, we have introduced Cys-X residues in human glutathione transferase (GST) M2-2, replacing Met-212 in the substrate-binding site. To achieve selectivity of the modifications, the Cys residues in the wild-type enzyme were replaced by Ala. A suite of simple substitutions resulted in a set of homologous Met derivatives ranging from normethionine to S-heptyl-cysteine. The chemical modifications were validated by HPLC and mass spectrometry. The derivatized mutant enzymes were assayed with alternative GST substrates representing diverse chemical reactions: aromatic substitution, epoxide opening, transnitrosylation, and addition to an ortho-quinone. The Cys substitutions had different effects on the alternative substrates and differentially enhanced or suppressed catalytic activities depending on both the Cys-X substitution and the substrate assayed. As a consequence, the enzyme specificity profile could be changed among the alternative substrates. The procedure lends itself to large-scale production of Cys-X modified protein variants.

A Systematic Review of Secondary Traumatic Stress and Compassion Fatigue in Teachers.

When teachers care for children with trauma histories, they are at risk of developing compassion fatigue (CF), or a reduced empathic capacity (Hupe and Stevenson in J Child Custody Res Issues Pract 16(4):364-386, 2019. https://doi.org/10.1080/15379418.2019.1663334). They may also develop secondary traumatic stress (STS), a secondary condition resulting from a person learning about details of a traumatic event experienced by someone in their care (Essary et al. in Kappa Delta Pi Record 56(3):116-121, 2020). While CF and STS have been studied widely in healthcare and mental health professionals (Baird and Kracen in Couns Psychol Q 19(2):181-188, 2006; Caringi et al. in Adv Sch Ment Health Promot 8(4):244-256, 2015. https://doi.org/10.1080/1754730X.2015.1080123; Cieslak et al. in Psychol Serv 11(1):75-86, 2014), STS and CF have been understudied in the teaching profession (Caringi et al., 2015; Christian-Brandt et al. in Child Abuse Neglect 110(3):104437, 2020; Hupe & Stevenson, 2019). As such, we sought to complete a systematic review of the literature to answer two questions: (1) To what extent are CF and STS being studied in teachers?; and (2) How have CF and STS been studied in teachers? Qualitative data analysis led to the emergence of four themes across all included studies: (1) conceptualization of CF and STS; (2) teachers are at risk of developing CF and STS; (3) varying approaches can mitigate the risk of CF and STS in teachers; and (4) there is limited research on CF and STS in teachers. Limitations and directions for future research and practice are described.

Engineering the enantioselectivity of glutathione transferase by combined active-site mutations and chemical modifications.

Based on the crystal structure of human glutathione transferase M1-1, cysteine residues were introduced in the substrate-binding site of a Cys-free mutant of the enzyme, which were subsequently alkylated with 1-iodoalkanes. By different combinations of site-specific mutations and chemical modifications of the enzyme the enantioselectivity in the conjugation of glutathione with the epoxide-containing substrates 1-phenylpropylene oxide and styrene-7,8-oxide were enhanced up to 9- and 10-fold. The results also demonstrate that the enantioselectivity can be diminished, or even reversed, by suitable modifications, which can be valuable under some conditions. The redesign of the active-site structure for enhanced or diminished enantioselectivities have divergent requirements for different epoxides, calling for a combinatorial approach involving alternative mutations and chemical modifications to optimize the enantioselectivity for a targeted substrate. This approach outlines a general method of great potential for fine-tuning substrate specificity and tailoring stereoselectivity of recombinant enzymes.

MBNL142 and MBNL143 gene isoforms, overexpressed in DM1-patient muscle, encode for nuclear proteins interacting with Src family kinases.

Myotonic dystrophy type-1 (DM1) is the most prevalent form of muscular dystrophy in adults. This disorder is an RNA-dominant disease, caused by expansion of a CTG repeat in the DMPK gene that leads to a misregulation in the alternative splicing of pre-mRNAs. The longer muscleblind-like-1 (MBNL1) transcripts containing exon 5 and the respective protein isoforms (MBNL142-43) were found to be overexpressed in DM1 muscle and localized exclusively in the nuclei. In vitro assays showed that MBNL142-43 bind the Src-homology 3 domain of Src family kinases (SFKs) via their proline-rich motifs, enhancing the SFK activity. Notably, this association was also confirmed in DM1 muscle and myotubes. The recovery, mediated by an siRNA target to Ex5-MBNL142-43, succeeded in reducing the nuclear localization of both Lyn and MBNL142-43 proteins and in decreasing the level of tyrosine phosphorylated proteins. Our results suggest an additional molecular mechanism in the DM1 pathogenesis, based on an altered phosphotyrosine signalling pathway.

Engineering GST M2-2 for high activity with indene 1,2-oxide and indication of an H-site residue sustaining catalytic promiscuity.

The substrate-binding H-site of human glutathione transferase (GST) M2-2 was subjected to iterative saturation mutagenesis in order to obtain an efficient enzyme with the novel epoxide substrate indene 1,2-oxide. Residues 10, 116, and 210 were targeted, and the activities with the alternative substrates, benzyl isothiocyanate and the prodrug azathioprine, undergoing divergent chemical reactions were monitored for comparison. In general, increased activities were found when the smaller residues Gly, Ser, and Ala replaced the original Thr210. The most active mutant T210G was further mutated at position 116, but no mutant showed enhanced catalytic activity. However, saturation mutagenesis of position 10 identified one double mutant T210G/I10C with 100-fold higher specific activity with indene 1,2-oxide than wild-type GST M2-2. This enhanced epoxide activity of 50 µmol min(-1) mg(-1) resulted primarily from an increased k(cat) value (70 s(-1)). The specific activity is 24-fold higher than that of wild-type GST M1-1, which is otherwise the most proficient GST enzyme with epoxide substrates. A second double mutant T210G/I10W displayed 30-fold increased activity with azathioprine, 0.56 µmol min(-1) mg(-1). In both double mutants, the replacement of Ile10 led to narrowed acceptance of alternative substrates. Ile10 is evolutionarily conserved in related class Mu GSTs. Conservation usually indicates preservation of a particular function, and in the Mu class, it would appear that the conserved Ile10 is not necessary to maintain catalytic functions but to prevent loss of broad substrate acceptance. In summary, our data underscore the facile transition between alternative substrate selectivity profiles in GSTs by a few mutations.

Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells.

Myotonic dystrophy (DM) is caused by a (CTG)(n) expansion in the 3'-untranslated region of DMPK gene. Mutant transcripts are retained in nuclear RNA foci, which sequester RNA binding proteins thereby misregulating the alternative splicing. Controversy still surrounds the pathogenesis of the DM1 muscle distress, characterized by myotonia, weakness and wasting with distal muscle atrophy. Eight primary human cell lines from adult-onset (DM1) and congenital (cDM1) patients, (CTG)(n) range 90-1800, were successfully differentiated into aneural-immature and contracting-innervated-mature myotubes. Morphological, immunohistochemical, RT-PCR and western blotting analyses of several markers of myogenesis indicated that in vitro differentiation-maturation of DM1 myotubes was comparable to age-matched controls. In all pathological muscle cells, (CTG)(n) expansions were confirmed by long PCR and RNA fluorescence in situ hybridization. Moreover, the DM1 myotubes showed the splicing alteration of insulin receptor and muscleblind-like 1 (MBNL1) genes associated with the DM1 phenotype. Considerable myotube loss and atrophy of 15-day-differentiated DM1 myotubes indicated activated catabolic pathways, as confirmed by the presence of apoptotic (caspase-3 activation, cytochrome c release, chromatin fragmentation) and autophagic (P62/LC3) markers. Z-VAD treatment significantly reduced the decrease in myonuclei number and in average width in 15-day-differentiated DM1 myotubes. We thus propose that the muscle wasting typical in DM1 is due to impairment of muscle mass maintenance-regeneration, through premature apoptotic-autophagic activation, rather than altered myogenesis.

Alternative mutations of a positively selected residue elicit gain or loss of functionalities in enzyme evolution.

All molecular species in an organism are connected physically and functionally to other molecules. In evolving systems, it is not obvious to what extent functional properties of a protein can change to selective advantage and leave intact favorable traits previously acquired. This uncertainty has particular significance in the evolution of novel pathways for detoxication, because an organism challenged with new xenobiotics in the environment may still require biotransformation of previously encountered toxins. Positive selection has been proposed as an evolutionary mechanism for facile adaptive responses of proteins to changing conditions. Here, we show, by saturation mutagenesis, that mutations of a hypervariable residue in human glutathione transferase M2-2 can differentially change the enzyme's substrate-activity profile with alternative substrates and, furthermore, enable or disable dissimilar chemical reactions. Crystal structures demonstrate that activity with epoxides is enabled through removal of steric hindrance from a methyl group, whereas activities with an orthoquinone and a nitroso donor are maintained in the variant enzymes. Given the diversity of cellular activities in which a single protein can be engaged, the selective transmutation of functional properties has general significance in molecular evolution.