Concomitant Occurrence of Peripheral Neuropathy and Vision Loss Due to Multivitamin Deficiency After Bariatric Surgery.

Vitamin deficiencies, especially after Bariatric surgery, are common and, when not properly addressed, can lead to debilitating complications. Bariatric procedures, to variable degrees, alter the anatomy and physiology of the gastrointestinal; this alteration makes these patients more susceptible to developing nutritional deficiencies. Peripheral neuropathy is one of the complications that can arise from nutritional deficiencies, and it can cause severe functional impairment. Vision loss is a relatively uncommon complication after weight loss procedure. Changes in the retinal nerve fiber layer, choroidal thickness, and visual fields due to hypovitaminosis result in nutritional optic neuropathy and retinopathy. The main retinal complication is nyctalopia (night blindness), which is caused by vitamin A deficiency. We present a case of concomitant peripheral neuropathy and vision loss secondary to reduced levels of multiple vitamins following gastric bypass surgery. This case highlights the need for regular vitamin level monitoring and appropriate replenishment in patients after bariatric surgery to prevent significant morbidities.

Coilin mediates m6A RNA methylation through phosphorylation of METTL3.

MicroRNAs (miRNAs) are a class of noncoding RNAs that regulate gene expression. An important step in miRNA biogenesis occurs when primary miRNAs are bound and cleaved by the microprocessor to generate precursor miRNAs. Regulation at this step is essential and one such regulator includes m6A RNA methylation, an RNA modification found on primary miRNAs that is installed by METTL3 and bound by hnRNPA2B1. Our lab has recently discovered that the Cajal body marker protein coilin also participates in miRNA biogenesis and hypothesized that coilin may be influencing miRNA biogenesis through m6A RNA methylation. Here we report that coilin suppression reduces m6A on primary Let7a and miR-21. We also found that coilin suppression reduced the protein expression of hnRNPA2B1 and METTL3. We observed an interaction between coilin and ectopically expressed METTL3 and found that coilin suppression reduced the nucleoplasmic portion of METTL3 and blunted ectopic METTL3 phosphorylation. Finally, coilin suppression disrupted the greater METTL3 complex with cofactors METTL14 and WTAP. Collectively, our work has uncovered a role for coilin in mediating m6A RNA methylation and provides an avenue by which coilin participates in miRNA biogenesis.

Examining the Perceptions Among Undergraduate Nursing Students Using Virtual Reality in a Community Course: A Mixed-Methods Explanatory Study.

PURPOSE: This study investigated undergraduate nursing students' perceptions of integrating virtual reality (VR) technology into a community nursing course, specifically home and environmental patient safety assessments. DESIGN: This study used a mixed-methods approach with an explanatory design. METHODS: Participants were recruited from a community health course. The students engaged in a VR simulation involving home visits using the vizHome platform. Quantitative and qualitative data were gathered using the Technology Acceptance Model (TAM) and the System Usability Scale (SUS). Semi-structured interviews were conducted. Quantitative data were collected through Qualtrics and secure Zoom connections for the interviews. The data were analyzed with SPSS and MAXQDA. FINDINGS: The results indicated that participants perceived VR as valuable and easy to use for learning home assessment skills. The System Usability Scale (SUS) score revealed room for improvement. Technical limitations were identified as challenges that must be addressed to enhance the user experience. CONCLUSION: Participants acknowledged VR's potential to supplement traditional learning methods, providing safe and realistic exposure to diverse home environments. While VR was seen as beneficial, it was not considered a replacement for actual home visits in community nursing education. CLINICAL EVIDENCE: This study provided clinical teaching evidence on the usability and student perceptions of VR in community courses.

The motivational beliefs and attitudes about writing of international students enrolled in online academic English classes during the COVID-19 pandemic.

Despite the growing attention to motivation, less is known about international students' motivational beliefs and attitudes about academic writing. In this study, we aimed to explore the motivational factors influencing international students' performance in academic English classes at a large public research university in the western United States. Specifically, we examined students' self-efficacy, goal orientation, beliefs, and affect for writing, along with their malleability, and their contributions to academic achievement in academic English writing classes. The sample comprised 97 students, predominantly from China, enrolled in online academic English courses. Exploratory factor analysis tended to extract more complex models of the motivational constructs than principal component analysis. Students' self-efficacy and enjoyment of writing significantly increased from the beginning to the end of the 10-week term, suggesting motivational factors' malleability. Hierarchical linear modeling revealed that students' self-efficacy at the beginning of the term positively predicted their final grades. However, logistic mixed modeling revealed that students who held stronger beliefs about writing as a means of exploring and expressing ideas had lower odds of passing. Our findings contribute to the understanding of international students' motivation in academic English settings in higher education and offers potential pedagogical interventions to enhance their academic success.

Coilin as a regulator of NF-kB mediated inflammation in preeclampsia.

The nuclear factor-Kappa B (NF-κB) pathway is a crucial mediator of inflammatory signaling. Aberrant activation of NF-κB is associated with several disorders including preeclampsia (PE). Many regulators of the NF-κB pathway have been identified, including microRNAs (miRNAs). Specifically, miR-517-3p targets mRNA encoding TNFAIP3 Interacting Protein 1 (TNIP1), an inhibitor of NF-κB signaling. Activation of NF-κB increases production of the cytokine TNF superfamily member 15 (TNFSF15), leading to the upregulation of anti-angiogenic soluble vascular endothelial growth factor receptor 1 (sFlt-1). We have previously observed that Cajal bodies (CBs), subnuclear domains, are associated with the chromosome 19 miRNA gene cluster (C19MC), which encodes miR-517-3p. We have also found that coilin, the CB marker protein, is a positive regulator of miRNA biogenesis. Here we report that coilin is a regulator of miR-517-3p, sFlt-1, TNIP1, TNFSF15 and NF-κB activation, and this regulation is influenced by hypoxia. We also report that coilin and CBs are induced in the reduced uterine perfusion pressure (RUPP) rat model of PE. Collectively, the data presented here implicate coilin as a novel regulator of NF-κB activation and sFlt-1 upregulation.

Usability of Microsoft Teams and Stream in Nursing Education.

This manuscript presents a technology acceptance and usability evaluation of an online software platform utilized for educating nursing students. Online nursing education is a complex and multifaceted process requiring nurse educators to be competent in technical informatics. Increasingly, nurse educators are asked to transform the traditional face-to-face classroom to include remote or hybrid experiences with little or no formal training. The pandemic may have created an atmosphere of stress and confusion while forcing a transition to an online learning environment. Recent trends in nursing education call for the increased use of technology that enhances the learning experience and mimic clinical practice. As the pandemic's restrictions are removed, nursing education will continue to require technological innovations. Variables measured during the study were usability (System Usability Scale), perceived usefulness, and perceived ease of use. The results of the study suggest participants perceived Microsoft Teams and Stream as useful and easy to use. In this situation, the use of technology positively supported the teaching-learning process.

Coilin enhances phosphorylation and stability of DGCR8 and promotes miRNA biogenesis.

MicroRNAs (miRNAs) are ∼22 nt small noncoding RNAs that control gene expression at the posttranscriptional level through translational inhibition and destabilization of their target mRNAs. The biogenesis of miRNAs involves a series of processing steps beginning with cropping of the primary miRNA transcript by the Microprocessor complex, which is composed of Drosha and DGCR8. Here we report a novel regulatory interaction between the Microprocessor components and coilin, the Cajal body (CB) marker protein. Coilin knockdown causes alterations in the level of primary and mature miRNAs, let-7a and miR-34a, and their miRNA targets, HMGA2 and Notch1, respectively. We also found that coilin knockdown affects the levels of DGCR8 and Drosha in cells with (HeLa) and without (WI-38) CBs. To further explore the role of coilin in miRNA biogenesis, we conducted a series of coimmunoprecipitation experiments using coilin and DGCR8 constructs, which revealed that coilin and DGCR8 can form a complex. Additionally, our results indicate that phosphorylation of DGCR8, which has been shown to increase protein stability, is impacted by coilin knockdown. Collectively, our results implicate coilin as a member of the regulatory network governing miRNA biogenesis.

The Cajal body protein coilin is a regulator of the miR-210 hypoxamiR and influences MIR210HG alternative splicing.

Hypoxia is a severe stressor to cellular homeostasis. At the cellular level, low oxygen triggers the transcription of a variety of genes supporting cell survival and oxygen homeostasis mediated by transcription factors, such as hypoxia-inducible factors (HIFs). Among many determinants dictating cell responses to hypoxia and HIFs are microRNAs (miRNAs). Cajal bodies (CBs), subnuclear structures involved in ribonucleoprotein biogenesis, have been recently proven to contribute to miRNA processing and biogenesis but have not been studied under hypoxia. Here, we show, for the first time, a hypoxia-dependent increase in CB number in WI-38 primary fibroblasts, which normally have very few CBs. Additionally, the CB marker protein coilin is upregulated in hypoxic WI-38 cells. However, the hypoxic coilin upregulation was not seen in transformed cell lines. Furthermore, we found that coilin is needed for the hypoxic induction of a well-known hypoxia-induced miRNA (hypoxamiR), miR-210, as well as for the hypoxia-induced alternative splicing of the miR-210 host gene, MIR210HG. These findings provide a new link in the physiological understanding of coilin, CBs and miRNA dysregulation in hypoxic pathology.

Cisplatin and phenanthriplatin modulate long-noncoding RNA expression in A549 and IMR90 cells revealing regulation of microRNAs, Wnt/ß-catenin and TGF-ß signaling.

The monofunctional platinum(II) complex, phenanthriplatin, acts by blocking transcription, but its regulatory effects on long-noncoding RNAs (lncRNAs) have not been elucidated relative to traditional platinum-based chemotherapeutics, e.g., cisplatin. Here, we treated A549 non-small cell lung cancer and IMR90 lung fibroblast cells for 24 h with either cisplatin, phenanthriplatin or a solvent control, and then performed microarray analysis to identify regulated lncRNAs. RNA22 v2 microRNA software was subsequently used to identify microRNAs (miRNAs) that might be suppressed by the most regulated lncRNAs. We found that miR-25-5p, -30a-3p, -138-5p, -149-3p, -185-5p, -378j, -608, -650, -708-5p, -1253, -1254, -4458, and -4516, were predicted to target the cisplatin upregulated lncRNAs, IMMP2L-1, CBR3-1 and ATAD2B-5, and the phenanthriplatin downregulated lncRNAs, AGO2-1, COX7A1-2 and SLC26A3-1. Then, we used qRT-PCR to measure the expression of miR-25-5p, -378j, -4516 (A549) and miR-149-3p, -608, and -4458 (IMR90) to identify distinct signaling effects associated with cisplatin and phenanthriplatin. The signaling pathways associated with these miRNAs suggests that phenanthriplatin may modulate Wnt/ß-catenin and TGF-ß signaling through the MAPK/ERK and PTEN/AKT pathways differently than cisplatin. Further, as some of these miRNAs may be subject to dissimilar lncRNA targeting in A549 and IMR90 cells, the monofunctional complex may not cause toxicity in normal lung compared to cancer cells by acting through distinct lncRNA and miRNA networks.

THE IMS PARADOX: A PERSPECTIVE ON STRUCTURAL ION MOBILITY-MASS SPECTROMETRY.

Studies of large proteins, protein complexes, and membrane protein complexes pose new challenges, most notably the need for increased ion mobility (IM) and mass spectrometry (MS) resolution. This review covers evolutionary developments in IM-MS in the authors' and key collaborators' laboratories with specific focus on developments that enhance the utility of IM-MS for structural analysis. IM-MS measurements are performed on gas phase ions, thus "structural IM-MS" appears paradoxical-do gas phase ions retain their solution phase structure? There is growing evidence to support the notion that solution phase structure(s) can be retained by the gas phase ions. It should not go unnoticed that we use "structures" in this statement because an important feature of IM-MS is the ability to deal with conformationally heterogeneous systems, thus providing a direct measure of conformational entropy. The extension of this work to large proteins and protein complexes has motivated our development of Fourier-transform IM-MS instruments, a strategy first described by Hill and coworkers in 1985 (Anal Chem, 1985, 57, pp. 402-406) that has proved to be a game-changer in our quest to merge drift tube (DT) and ion mobility and the high mass resolution orbitrap MS instruments. DT-IMS is the only method that allows first-principles determinations of rotationally averaged collision cross sections (CSS), which is essential for studies of biomolecules where the conformational diversities of the molecule precludes the use of CCS calibration approaches. The Fourier transform-IM-orbitrap instrument described here also incorporates the full suite of native MS/IM-MS capabilities that are currently employed in the most advanced native MS/IM-MS instruments. © 2020 John Wiley & Sons Ltd. Mass Spec Rev.

Molecular determinants that govern scaRNA processing by Drosha/DGCR8.

The Cajal body (CB) is a subnuclear domain that participates in the biogenesis of many different types of ribonucleoproteins (RNPs), including small nuclear RNPs (snRNPs), small Cajal body-specific RNPs (scaRNPs) and telomerase. Most scaRNAs, the RNA component of scaRNPs, accumulate in CBs. However, there are three scaRNAs (scaRNA 2, 9, and 17) that are known to be processed into small, nucleolar-enriched fragments. Evidence suggests that these fragments are packaged into a new class of RNPs, called regulatory RNPs (regRNPs), and may modify small nucleolar RNP (snoRNP) activity, thus playing a role in rRNA modification. However, the mechanism by which these fragments are produced is unknown. Previous work has reported the involvement of Drosha and DGCR8 in the cleavage of primary-scaRNA9. Here, we expand on that knowledge by identifying sequence elements necessary for the efficient production of these RNA fragments and demonstrate that primary scaRNA 2 and 17 are also processed by the Drosha-DGCR8 complex. Collectively, our work establishes new factors in the scaRNP biogenesis pathway and adds to the ever-expanding list of noncanonical functions for the microprocessor complex.

First-Principles Collision Cross Section Measurements of Large Proteins and Protein Complexes.

Rotationally averaged collision cross section (CCS) values for a series of proteins and protein complexes ranging in size from 8.6 to 810 kDa are reported. The CCSs were obtained using a native electrospray ionization drift tube ion mobility-Orbitrap mass spectrometer specifically designed to enhance sensitivity while having high-resolution ion mobility and mass capabilities. Periodic focusing (PF)-drift tube (DT)-ion mobility (IM) provides first-principles determination of the CCS of large biomolecules that can then be used as CCS calibrants. The experimental, first-principles CCS values are compared to previously reported experimentally determined and computationally calculated CCS using projected superposition approximation (PSA), the Ion Mobility Projection Approximation Calculation Tool (IMPACT), and Collidoscope. Experimental CCS values are generally in agreement with previously reported CCSs, with values falling within ∼5.5%. In addition, an ion mobility resolution (CCS centroid divided by CCS fwhm) of ∼60 is obtained for pyruvate kinase (MW ∼ 233 kDa); however, ion mobility resolution for bovine serum albumin (MW ∼ 68 kDa) is less than ∼20, which arises from sample impurities and underscores the importance of sample quality. The high resolution afforded by the ion mobility-Orbitrap mass analyzer provides new opportunities to understand the intricate details of protein complexes such as the impact of post-translational modifications (PTMs), stoichiometry, and conformational changes induced by ligand binding.

Synergistic interactions between Cajal bodies and the miRNA processing machinery.

Cajal bodies (CBs) are subnuclear domains involved in the formation of ribonucleoproteins (RNPs) including small nuclear RNPs (snRNPs). CBs associate with specific gene loci, which impacts expression and provides a platform for the biogenesis of the nascent transcripts emanating from these genes. Here we report that CBs can associate with the C19MC microRNA (miRNA) gene cluster, which suggests a role for CBs in the biogenesis of animal miRNAs. The machinery involved in the formation of miRNAs includes the Drosha/DGCR8 complex, which processes primary-miRNA to precursor miRNA. Further processing of precursor miRNA by Dicer and other components generates mature miRNA. To test if CBs influence the expression and formation of miRNAs, we examined two representative miRNAs (miR-520 h and let-7a) in conditions that disrupt CBs. CB disruption correlates with alterations in the level of primary and mature miRNA and the let-7a mRNA target, HMGA2. We have also found that the processing of some small CB-specific RNAs (scaRNAs) is directly mediated by the Drosha/DGCR8 complex. ScaRNAs form scaRNPs, which play an important role in snRNP formation. Collectively, our results demonstrate that CBs and the miRNA processing machinery functionally interact and together contribute to the biogenesis of miRNAs and snRNPs.

Do Children Classified With Specific Language Impairment Have a Learning Disability in Writing? A Meta-Analysis.

In this meta-analysis, we examined whether children classified with specific language impairment (SLI) experience difficulties with writing. We included studies comparing children with SLI to (a) typically developing peers matched on age (k = 39 studies) and (b) typically developing younger peers with similar language capabilities (k = six studies). Children classified with SLI scored lower on writing measures than their typically developing peers matched on age (g = -0.97) when all writing scores in a study were included in the analysis. This same pattern occurred for specific measures of writing: quality (g = -0.92), output (g = -1.00), grammar (g = -0.68), vocabulary (g = -0.68), and spelling (g = -1.17). A moderator analysis revealed that differences in the writing scores of children classified with SLI and typically developing peers matched on age were not as large, but were still statistically significant, when assessment involved a contrived response format (vs. measured based on students' writing), researcher-created measures (vs. norm-referenced tests), or SLI included just children with a speech disorder (vs. children with a language disorder). Children classified with SLI further scored lower on writing than typically developing peers with similar language capabilities (g = -0.47). We concluded that children with SLI experience difficulties with writing.

Tracking the Structural Evolution of 4-Aminobenzoic Acid in the Transition from Solution to the Gas Phase.

Here, cryogenic ion mobility-mass spectrometry (cryo-IM-MS) is used to investigate intracluster proton transfer reactions of 4-aminobenzoic acid during the transition from solution to the gas phase. Previous studies have shown that protonation of the amine group of 4-aminobenzoic acid (4-ABAH+) is favored in solution (N-protomer), whereas protonation of the carboxylic acid group is favored in the gas phase (O-protomer). Results from cryo-IM-MS (80 K) studies of hydrated 4-ABAH+ ions, 4-ABAH+(H2O)n, are interpreted as evidence that the proton transfer reaction occurs through a water bridge at n = 6 connecting the -NH3+ and -COOH groups, that is, a Grotthuss mechanism. The weak binding energy of water molecules imposes limits for obtaining first-principles collisional cross sections (CCSs) of hydrated ions; consequently, candidate structures for 4-ABAH+(H2O)0-6 ions are derived by correlating experimental arrival-time distributions to theoretically determined CCSs. To our knowledge, these are the first first-principles determinations of CCS for hydrated ions. Apolar cosolvents, particularly acetonitrile, have been postulated to inhibit proton transfer by blocking the Grotthuss mechanism, but our data suggest that acetonitrile simply stabilizes the ammonium ion.

Alteration of 28S rRNA 2'-O-methylation by etoposide correlates with decreased SMN phosphorylation and reduced Drosha levels.

The most common types of modification in human rRNA are pseudouridylation and 2'-O ribose methylation. These modifications are performed by small nucleolar ribonucleoproteins (snoRNPs) which contain a guide RNA (snoRNA) that base pairs at specific sites within the rRNA to direct the modification. rRNA modifications can vary, generating ribosome heterogeneity. One possible method that can be used to regulate rRNA modifications is by controlling snoRNP activity. RNA fragments derived from some small Cajal body-specific RNAs (scaRNA 2, 9 and 17) may influence snoRNP activity. Most scaRNAs accumulate in the Cajal body - a subnuclear domain - where they participate in the biogenesis of small nuclear RNPs, but scaRNA 2, 9 and 17 generate nucleolus-enriched fragments of unclear function, and we hypothesize that these fragments form regulatory RNPs that impact snoRNP activity and modulate rRNA modifications. Our previous work has shown that SMN, Drosha and various stresses, including etoposide treatment, may alter regulatory RNP formation. Here we demonstrate that etoposide treatment decreases the phosphorylation of SMN, reduces Drosha levels and increases the 2'-O-methylation of two sites within 28S rRNA. These findings further support a role for SMN and Drosha in regulating rRNA modification, possibly by affecting snoRNP or regulatory RNP activity.

Hydration of Guanidinium Ions: An Experimental Search for Like-Charged Ion Pairs.

Guanidinium ions (GdmH+) are reported to form stable complexes (GdmH+/GdmH+) in aqueous solution despite strong repulsive interactions between the like-charged centers. These complexes are thought to play important roles in protein folding, membrane penetration, and formation of protein dimers. Although GdmH+ ions are weakly hydrated, semiempirical calculations provide evidence that these like-charged complexes are stabilized by water molecules, which serve important structural and energetic roles. Specifically, water molecules bridge between the GdmH+ ions of GdmH+/GdmH+ complexes as well as complexes involving the guanidinium side chains of arginine. Potential biological significances of like-charged complexes have been largely confirmed by ab initio molecular dynamics simulations and indirect experimental evidence. We report cryo-ion mobility-mass spectrometry results for the GdmH+/GdmH+ ion pair confined in a nanodroplet- the first direct experimental observation of this like-charged complex. A second like-charged complex, described as a water-mediated complex involving GdmH+ and H3O+, was also observed.

Why Children With Dyslexia Struggle With Writing and How to Help Them.

Purpose: Children with dyslexia often have related writing difficulties. In the simple view of writing model, high-quality writing depends on good transcription skills, working memory, and executive function-all of which can be difficult for children with dyslexia and result in poor spelling and low overall writing quality. In this article, we describe the challenges of children with dyslexia in terms of the simple view of writing and instructional strategies to increase spelling and overall writing quality in children with dyslexia. Method: For spelling strategies, we conducted systematic searches across 2 databases for studies examining the effectiveness of spelling interventions for students with dyslexia as well as including studies from 2 meta-analyses. To locate other instructional practices to increase writing quality (e.g., handwriting and executive function), we examined recent meta-analyses of writing and supplemented that by conducting forward searches. Results: Through the search, we found evidence of effective remedial and compensatory intervention strategies in spelling, transcription, executive function, and working memory. Some strategies included spelling using sound-spellings and morphemes and overall quality using text structure, sentence combining, and self-regulated strategy development. Conclusions: Many students with dyslexia experience writing difficulty in multiple areas. However, their writing (and even reading) skills can improve with the instructional strategies identified in this article. We describe instructional procedures and provide links to resources throughout the article.

Altered dynamics of scaRNA2 and scaRNA9 in response to stress correlates with disrupted nuclear organization.

Small Cajal body-specific RNAs (scaRNAs) are part of small Cajal body-specific ribonucleoproteins (scaRNPs) that modify small nuclear RNA (snRNA) in Cajal bodies (CBs). Several scaRNAs (scaRNA 2, 9 and 17) have been found to generate smaller, nucleolus-enriched fragments. We hypothesize that the fragments derived from scaRNA 2, 9 and 17 form regulatory RNPs that influence the level of modifications within rRNA by altering small nucleolar RNP (snoRNP) activity. Here we show that external factors such as DNA damaging agents can alter the scaRNA9 full length to processed fragment ratio. We also show that full-length scaRNA2 levels are likewise impacted by DNA damage, which correlates with the disruption of SMN, coilin and WRAP53 co-localization in CBs. The dynamics of scaRNA9 were also shown to be affected by Drosha levels, which suggests that this protein may participate in the biogenesis and processing of this non-coding RNA. Identification of factors that contribute to scaRNA 2, 9 and 17 processing may facilitate an assessment of how external stress can lead to changes in rRNA modifications.