bluuebunny/crossref_metadata_2025_split huggingface dataset:
Equitable practices in school mental health
Authors: DeBoer Jacob L. , Allouche Sam F. , Vasquez Jennifer I. , Rhodes Judith 
Published: February 2022

AbstractGiven the prevalence and negative outcomes associated with untreated mental health concerns, there is an urgent need for schools to enhance their capacity to prevent, identify, and address these needs. This is particularly important for students from culturally and linguistically diverse backgrounds, who are more likely to be underserved and face barriers to mental health treatment. Current methods of addressing mental health at school are reactive and insufficient, failing to address the needs of all students. Without an explicit focus on dismantling systemic racism and inequity within schools, our society will continue to underserve those already disenfranchised. As such, this paper outlines the challenges as well as the importance of incorporating culture into a conceptualization of universal mental health in schools. We include state‐, district‐, and school‐level solutions in this conceptualization. Specifically, we propose sample solutions to the implementation of equity‐focused, comprehensive mental health practices within a multitiered system of support, focusing predominantly on universal (i.e., tier 1 level) practices.
Bronchoscopy in the management of children from developing countries undergoing congenital heart surgery
Authors: Schnapper Michael , Dalal Ilan , Mandelberg Avigdor , Raucher Sternfeld Alona , Sasson Lior , Armoni Domany Keren 
Published: February 2022

AbstractObjectivesTo study the clinical characteristics and impact of bronchoscopy in children from developing countries, referred for cardiac surgery, through the "Save a Child's Heart" (SACH) organization.MethodsWe performed a retrospective hospital‐chart review of SACH children (0–18 years old) referred between 2006 and 2021 who underwent fiberoptic bronchoscopy. We examined demographics, congenital‐heart‐disease (CHD) types, bronchoscopy's indications and findings, subsequent recommendations, number of ventilation, and intensive‐care‐unit days. The primary outcome was percent changes in management and diagnosis, following the bronchoscopy. We included a control group matched‐for‐age and CHD type, who did not undergo bronchoscopy.ResultsWe performed 82 bronchoscopies in 68 children: 18 (26.5%) preoperatively; 46 (67.6%) postoperatively; and four (5.9%) both. The most prevalent CHDs were Tetralogy‐of‐Fallot (27.9%) and ventricular‐septal‐defect (19.1%). The main indications were persistent atelectasis (41%) and mechanical ventilation/weaning difficulties (27.9%). Bronchoscopic evaluations revealed at least one abnormality in 51/68 (75%) children. The most common findings were external airway compression (23.5%), bronchomalacia (19.1%), and mucus secretions (14.7%). Changes in management were made in 35 (51.4%) cases, with a major change made in 14/35 (40%) children. Compared to the control group, the children undergoing bronchoscopy were both ventilated longer (median 6 vs. 1.5 days, p < 0.0001) and stayed longer in the intensive care unit (median 1.5 vs. 18.5 days, p < 0.0001).ConclusionA bronchoscopy is an important tool in the diagnosis and management of the unique group of children from developing countries with CHD referred for cardiac surgery. The results of our study, reveal a more complicated clinical course in children requiring bronchoscopy compared to controls.
Ultra‐Thermostability of Spatially Confined and Fully Protected Perovskite Nanocrystals by In Situ Crystallization
Authors: Li Qinyi , Shen Dongyang , Luo Chengzhao , Zheng Zhishuai , Xia Wenlin , Ma Wenchen , Li Jie , Yang Yixing , Chen Song , Chen Yu 
Published: February 2022

AbstractAlthough all‐inorganic perovskite materials present multiple fascinating optical properties, their poor stability undermines their potential application in the field of multi‐color display. Herein, spatially confined CsPbBr3 nanocrystals are in situ crystallized within uniform mesoporous SiO2 nanospheres (MSNs) to regulate their size distribution, passivate their surface defects, shield them from water/oxygen, and more importantly, enhance their thermotolerance. As a result, the remnant PL intensity of the prepared spatially confined perovskite (CsPbBr3) nanocrystals by in situ crystallization within uniform mesoporous SiO2 nanospheres (SCP@MSNs) powders can be maintained over 98% of its initial value even after being immersed in harsh conditions (0.1 m HCl or 0.1 m NaOH) for 60 days. Furthermore, the prepared SCP@MSNs‐PDMS film demonstrates astonishing thermostability by maintaining almost consistent room temperature PL intensities after continuous heating–cooling cycles between 200 and 25 °C, which would greatly improve its processability during potential industrial manufacturing. The fabricated LCD backlit based on SCP@MSNs covers 124% of NTSC standard and 95.6% of Rec. 2020 standard, indicating its great potential in practical display field.
Towards an AI‐driven business development framework: A multi‐case study
Authors: John Meenu Mary , Olsson Helena Holmström , Bosch Jan 
Published: February 2022

AbstractArtificial intelligence (AI) and the use of machine learning (ML) and deep learning (DL) technologies are becoming increasingly popular in companies. These technologies enable companies to leverage big quantities of data to improve system performance and accelerate business development. However, despite the appeal of ML/DL, there is a lack of systematic and structured methods and processes to help data scientists and other company roles and functions to develop, deploy and evolve models. In this paper, based on multi‐case study research in six companies, we explore practices and challenges practitioners experience in developing ML/DL models as part of large software‐intensive embedded systems. Based on our empirical findings, we derive a conceptual framework in which we identify three high‐level activities that companies perform in parallel with the development, deployment and evolution of models. Within this framework, we outline activities, iterations and triggers that optimize model design as well as roles and company functions. In this way, we provide practitioners with a blueprint for effectively integrating ML/DL model development into the business to achieve better results than other (algorithmic) approaches. In addition, we show how this framework helps companies solve the challenges we have identified and discuss checkpoints for terminating the business case.
Protective effect of nimbolide against streptozotocin induced gestational diabetes mellitus in rats via alteration of inflammatory reaction, oxidative stress, and gut microbiota
Authors: Ma Yifei , Xu Shan , Meng Juan , Li Lu 
Published: February 2022

AbstractBackgroundGestational diabetes mellitus (GDM) is a significant pregnancy‐related condition, which showed effect on the development of fetal. Anti‐inflammatory and antioxidant therapy commonly used for the treatment of GDM. Nimbolide already confirmed their anti‐inflammatory and anti‐oxidant effect against various animal disease model. Our objective in this research is to investigate the protective effect of nimbolide against STZ induced GDM and elucidate the mechanism.MethodsIn this experimental study, pregnant female Wistar rats were used and STZ (40 mg/kg) was used to induce the GDM. Blood glucose level (BGL), body weight and plasma insulin were assessed at regular time (gestational day 0, 9, and 18). Water intake, food intake, fecal and urine output were also estimated. In the female rats, hemoglobin (Hb), glycalated hemoglobin (HbA1c), hepatic glycogen, fructosamine, adiponectin, leptin, lipid, antioxidant and inflammatory cytokines parameters were estimated. In the fetuses, the fetues weight, implementation loss, and fetal weight were estimated. At the completion of the protocol, biochemical parameters were calculated. Gut microbiota was estimated in end of the study.ResultsNimbolide treatment significantly (p < .001) improved the fetuses level and suppressed the fetal weight and implantation loss. Nimbolide treatment significantly (p < .001) suppressed the BGL and enhanced the body weight, insulin level. Nimbolide treatment suppressed the water intake, food intake, urinary and fecal output. Nimbolide significantly (p < .001) suppressed the fructosamine, leptin and enhanced the adiponectin level. Nimbolide treatment significantly (p < .001) decreased the malonaldehyde (MDA) level and boosted the total antioxidant capacity (TAC), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S‐transferase (GST) and catalase (CAT); suppressed the level of TNF‐α, IL‐1β, IL‐6, and boosted the level of IL‐10. Furthermore, nimbolide treatment reversed the gut microbiota alteration induced via STZ in female rats. At the phylum level, the Firmicutes and Bacteroidetes relative abundance was altered via nimbolide treatment. The ratio of F/B boosted in GDM group and nimbolide treatment significantly (p < .001) suppressed. Nimbolide considerably suppressed the firmicutes and enhanced the Bacteroidetes, CAG‐352, Lacnospirace.ConclusionBased on the findings, we may conclude that nimbolide protects the pregnant rats from GDM via alteration of inflammation, oxidative stress, and gut microbiota.
In Vitro Models for the Evaluation of Antisense Oligonucleotides in Skin
Authors: Bremer Jeroen , van den Akker Peter C. 
Published: February 2022

AbstractThe genodermatosis dystrophic epidermolysis bullosa (DEB) is caused by mutations in the COL7A1 gene which encodes type VII collagen (C7). In the cutaneous basement membrane zone, C7 secures attachment of the epidermal basal keratinocyte to the papillary dermis by means of anchoring fibril formation. The complete absence of these anchoring fibrils leads to severe blistering of skin and mucosa upon the slightest friction and early mortality. To date, although preclinical advances toward therapy are promising, treatment for the disease is merely symptomatic. Therefore, research into novel therapeutics is warranted.Antisense oligonucleotide (ASO)-mediated exon skipping is such a therapy. Clinical examination of naturally occurring exon skipping suggested that this mechanism could most likely benefit the most severely affected patients. The severe form of DEB is caused by biallelic null mutations. Exon skipping aims to bind an ASO to the mutated exon of the pre-mRNA in the cell nucleus. Thereby, the ASO inhibits the recognition of the mutated exon by the splicing machinery, and as a result, the mutated exon is spliced out from the mRNA with its surrounding introns, i.e., it is skipped. Here, we describe in vitro methods to evaluate ASO-mediated exon skipping in a preclinical setting.
In Vitro Delivery of PMOs in Myoblasts by Electroporation
Authors: Goossens Remko , Aartsma-Rus Annemieke 
Published: February 2022

AbstractAntisense oligonucleotides (AONs) are small synthetic molecules of therapeutic interest for a variety of human disease. Their ability to bind mRNA and affect its splicing gives AONs potential use for exon skipping therapies aimed at restoring the dystrophin transcript reading frame for Duchenne muscular dystrophy (DMD) patients. The neutrally charged phosphorodiamidate morpholino oligomers (PMOs) are a stable and relatively nontoxic AON modification. To assess exon skipping efficiency in vitro, it is important to deliver them to target cells. Here, we describe a method for the delivery of PMOs to myoblasts by electroporation. The described protocol for the Amaxa 4D X unit nucleofector system allows efficient processing of 16 samples in one nucleocuvette strip, aiding in high-throughput PMO efficacy screens.
Rapid Determination of MBNL1 Protein Levels by Quantitative Dot Blot for the Evaluation of Antisense Oligonucleotides in Myotonic Dystrophy Myoblasts
Authors: Moreno Nerea , González-Martínez Irene , Artero Rubén , Cerro-Herreros Estefanía 
Published: February 2022

AbstractWestern blot assays are not adequate for high-throughput screening of protein expression because it is an expensive and time-consuming technique. Here we demonstrate that quantitative dot blots in plate format are a better option to determine the absolute contents of a given protein in less than 48 h. The method was optimized for the detection of the Muscleblind-like 1 protein in patient-derived myoblasts treated with a collection of more than 100 experimental oligonucleotides.
Evaluation of Exon Skipping and Dystrophin Restoration in In Vitro Models of Duchenne Muscular Dystrophy
Authors: López-Martínez Andrea , Soblechero-Martín Patricia , Arechavala-Gomeza Virginia 
Published: February 2022

AbstractSeveral exon skipping antisense oligonucleotides (eteplirsen, golodirsen, viltolarsen, and casimersen) have been approved for the treatment of Duchenne muscular dystrophy, but many more are in development targeting an array of different DMD exons. Preclinical screening of the new oligonucleotide sequences is routinely performed using patient-derived cell cultures, and evaluation of their efficacy may be performed at RNA and/or protein level. While several methods to assess exon skipping and dystrophin expression in cell culture have been developed, the choice of methodology often depends on the availability of specific research equipment.In this chapter, we describe and indicate the relevant bibliography of all the methods that may be used in this evaluation and describe in detail the protocols routinely followed at our institution, one to evaluate the efficacy of skipping at RNA level (nested PCR) and the other the restoration of protein expression (myoblot), which provide good results using equipment largely available to most research laboratories.
Generation of Human iPSC-Derived Myotubes to Investigate RNA-Based Therapies In Vitro
Authors: Herrero-Hernandez Pablo , Bergsma Atze J. , Pijnappel W. W. M. Pim 
Published: February 2022

AbstractAlternative pre-mRNAsplicing can be cell-type specific and results in the generation of different protein isoforms from a single gene. Deregulation of canonical pre-mRNAsplicing by disease-associated variants can result in genetic disorders. Antisense oligonucleotides (AONs) offer an attractive solution to modulate endogenous gene expression through alteration of pre-mRNAsplicing events. Relevant in vitro models are crucial for appropriate evaluation of splicing modifying drugs. In this chapter, we describe how to investigate the splicing modulating activity of AONs in an in vitro skeletal muscle model, applied to Pompe disease. We also provide a detailed description of methods to visualize and analyze gene expression in differentiated skeletal muscle cells for the analysis of muscledifferentiation and splicing outcome. The methodology described here is relevant to develop treatment options using AONs for other genetic muscle diseases as well, including Duchenne muscular dystrophy, myotonic dystrophy, and facioscapulohumeral muscular dystrophy.
Eye on a Dish Models to Evaluate Splicing Modulation
Authors: Hau Kwan-Leong , Lane Amelia , Guarascio Rosellina , Cheetham Michael E. 
Published: February 2022

AbstractInherited retinal dystrophies, such as Leber congenital amaurosis, Stargardt disease, and retinitis pigmentosa, are characterized by photoreceptor dysfunction and death and currently have few treatment options. Recent technological advances in induced pluripotent stem cell (iPSC) technology and differentiation methods mean that human photoreceptors can now be studied in vitro. For example, retinal organoids provide a platform to study the development of the human retina and mechanisms of diseases in the dish, as well as being a potential source for cell transplantation. Here, we describe differentiation protocols for 3D cultures that produce retinal organoids containing photoreceptors with rudimentary outer segments. These protocols can be used as a model to understand retinal disease mechanisms and test potential therapies, including antisense oligonucleotides (AONs) to alter gene expression or RNA processing. This “retina in a dish” model is well suited for use with AONs, as the organoids recapitulate patient mutations in the correct genomic and cellular context, to test potential efficacy and examine off-target effects on the translational path to the clinic.
Antisense RNA Therapeutics: A Brief Overview
Authors: Arechavala-Gomeza Virginia , Garanto Alejandro 
Published: February 2022

AbstractNucleic acid therapeutics is a growing field aiming to treat human conditions that has gained special attention due to the successful development of mRNA vaccines against SARS-CoV-2. Another type of nucleic acid therapeutics is antisense oligonucleotides, versatile tools that can be used in multiple ways to target pre-mRNA and mRNA. While some years ago these molecules were just considered a useful research tool and a curiosity in the clinical market, this has rapidly changed. These molecules are promising strategies for personalized treatments for rare genetic diseases and they are in development for very common disorders too. In this chapter, we provide a brief description of the different mechanisms of action of these RNA therapeutic molecules, with clear examples at preclinical and clinical stages.
Delivery of Antisense Oligonucleotides to the Mouse Retina
Authors: Garanto Alejandro 
Published: February 2022

AbstractThe eye is the organ in charge of vision and, given its properties, has become an excellent organ to test genetic therapies, including antisense oligonucleotide (AON) technology. In fact, the first AON receiving FDA and EMA approval was meant to treat an eye condition. Currently, dozens of clinical trials are being conducted for a variety of subtypes of inherited retinal disease. Although most of them are based on gene augmentation therapies, a phase 3 and two phase 1/2 clinical trials using AONs are ongoing. Since the retina is a layered structure of nondividing cells, obtaining human retinal tissue and expanding it in the lab is not possible, unless induced pluripotent stem cell technology is used. Mouse models have helped to elucidate the function of many genes, and the retinal structure is quite similar to that of humans. Thus, drug delivery to the mouse eye can provide valuable information for further optimization of therapies. In this chapter, the protocol for intravitreal injections of AONs is described in detail.
Preclinical Evaluation of the Renal Toxicity of Oligonucleotide Therapeutics in Mice
Authors: Echevarría Lucía , Goyenvalle Aurelie 
Published: February 2022

AbstractAntisense oligonucleotides (ASO) therapeutics hold great promise for the treatment of numerous diseases, and several ASO drugs have now reached market approval, confirming the potential of this approach. However, some candidates have also failed, due to limited biodistribution/uptake and poor safety profile. In pursuit of better delivery and higher cellular uptake, ASO are being optimized, and new chemistries are developed or conjugated with various ligands. While these developments may lead to candidates with higher potency, it is important to keep the safety aspects in sight and screen for potential toxicity in early phases of preclinical development to avoid subsequent failure in clinical development. Our understanding of ASO-mediated toxicity keeps improving with increased preclinical and clinical data available. In this chapter, we will focus on the assessment of renal toxicity in mice and describe methods to measure the levels of general urinary biomarkers as well as acute kidney injury biomarkers following ASO treatment.
Determination of Optimum Ratio of Cationic Polymers and Small Interfering RNA with Agarose Gel Retardation Assay
Authors: Aydin Omer , Kanarya Dilek , Yilmaz Ummugulsum , Tunç Cansu Ümran 
Published: February 2022

AbstractNanomaterials have aroused attention in the recent years for their high potential for gene delivery applications. Most of the nanoformulations used in gene delivery are positively charged to carry negatively charged oligonucleotides. However, excessive positively charged carriers are cytotoxic. Therefore, the complexed oligonucleotide/nanoparticles should be well-examined before the application. In that manner, agarose gel electrophoresis, which is a basic method utilized for separation, identification, and purification of nucleic acid molecules because of its poriferous nature, is one of the strategies to determine the most efficient complexation rate. When the electric field is applied, RNA fragments can migrate through anode due to the negatively charged phosphate backbone. Because RNA has a uniform mass/charge ratio, RNA molecules run in agarose gel proportional according to their size and molecular weight. In this chapter, the determination of complexation efficiency between cationic polymer carriers and small interfering RNA (siRNA) cargos by using agarose gel electrophoresis is described. siRNA/cationic polymer carrier complexes are placed in an electric field and the charged molecules move through the counter-charged electrodes due to the phenomenon of electrostatic attraction. Nucleic acid cargos are loaded to cationic carriers via the electrostatic interaction between positively charged amine groups (N) of the carrier and negatively charged phosphate groups (P) of RNA. The N/P ratio determines the loading efficiency of the cationic polymer carrier. In here, the determination of N/P ratio, where the most efficient complexation occurs, by exposure to the electric field with a gel retardation assay is explained.
Development and Use of Cellular Systems to Assess and Correct Splicing Defects
Authors: Suárez-Herrera Nuria , Tomkiewicz Tomasz Z. , Garanto Alejandro , Collin Rob W. J. 
Published: February 2022

AbstractA significant proportion of mutations underlying genetic disorders affect pre-mRNA splicing, generally causing partial or total skipping of exons, and/or inclusion of pseudoexons. These changes often lead to the formation of aberrant transcripts that can induce nonsense-mediated decay, and a subsequent lack of functional protein. For some genetic disorders, including inherited retinal diseases (IRDs), reproducing splicing dynamics in vitro is a challenge due to the specific environment provided by, e.g. the retinal tissue, cells of which cannot be easily obtained and/or cultured. Here, we describe how to engineer splicing vectors, validate the reliability and reproducibility of alternative cellular systems, assess pre-mRNA splicing defects involved in IRD, and finally correct those by using antisense oligonucleotide-based strategies.
Introduction: The Pandemic of Argumentation
Authors: Oswald Steve , Lewiński Marcin , Greco Sara , Villata Serena 
Published: February 2022

AbstractThis chapter introduces the volume and contextualises its scope, which covers communicative aspects of the current COVID-19 pandemic as well as the epidemic of misinformation from the perspective of argumentation theory. Argumentation theory is uniquely placed to understand and account for the challenges of public reason as expressed through argumentative discourse. The book thus focuses on the extent to which the forms, norms and functions of public argumentation have changed in the face of the COVID-19 pandemic. This question is investigated along descriptive, normative and prescriptive research lines at the core of the COST Action project CA 17132: European network for Argumentation and Public PoLicY analysis (APPLY). Contributions are divided into three groups, which (i) examine various features and aspects of public and institutional discourse about the COVID-19 pandemic, (ii) scrutinise the way health policies have been discussed, debated, attacked and defended in the public sphere, and (iii) consider a range of measures meant to improve the quality of public discourse, and public deliberation in particular, in such a way that concrete proposals for argumentative literacy are brought to light.