Enhanced HbF reactivation by multiplex mutagenesis of thalassemic CD34+ cells in vitro and in vivo.

Thalassemia or sickle cell patients with hereditary persistence of fetal hemoglobin (HbF) have an ameliorated clinical phenotype and, in some cases, can achieve transfusion independence. Inactivation via genome editing of γ-globin developmental suppressors, such as BCL11A or LRF/ZBTB7A, or of their binding sites, have been shown to significantly increase expression of endogenous HbF. To broaden the therapeutic window beyond a single-editing approach, we have explored combinations of cis- and trans-editing targets to enhance HbF reactivation. Multiplex mutagenesis in adult CD34+ cells was well tolerated and did not lead to any detectable defect in the cells' proliferation and differentiation, either in vitro or in vivo. The combination of 1 trans and 1 cis mutation resulted in high editing retention in vivo, coupled with almost pancellular HbF expression in NBSGW mice. The greater in vivo performance of this combination was also recapitulated using a novel helper-dependent adenoviral-CRISPR vector (HD-Ad-dualCRISPR) in CD34+ cells from ß-thalassemia patients transplanted to NBSGW mice. A pronounced increase in HbF expression was observed in human red blood cells in mice with established predominant ß0/ß0-thalassemic hemopoiesis after in vivo injection of the HD-Ad-dualCRISPR vector. Collectively, our data suggest that the combination of cis and trans fetal globin reactivation mutations has the potential to significantly increase HbF both totally and on a per cell basis over single editing and could thus provide significant clinical benefit to patients with severe ß-globin phenotype.

High body mass index and sleep problems during pregnancy: A meta-analysis and meta-regression of observational studies.

Despite the well-established correlation of weight and sleeping problems, little is known about the nature of the association. The present study examined whether pregnant women with high body mass index have a risk of developing sleep problems, and identified any covariates that affect this relationship. We systematically searched electronic databases, specialized journals, various clinical trial registries, grey literature databases and the reference list of the identified studies. All observational studies were obtained from inception until 9 August 2020. The Newcastle-Ottawa Scale was adopted to assess the quality of studies. Stata software was used to conduct meta-analysis and meta-regression. Forty-six observational studies involving 2,240,804 participants across 16 countries were included. Quality assessment scores ranged from 4 to 10 (median = 6). Meta-analyses revealed that the risk of sleep apnea, habitual snoring, short sleep duration and poor sleep quality is increased in pregnant women with high body mass index, but not for daytime sleepiness, insomnia or restless legs syndrome. Subgroup differences were detected on body mass index between different regions, nature of population, year of publication, age group and study quality. Random-effects meta-regression analyses showed that year and quality of publication were covariates on the relationships between pre-pregnant body mass index and sleep apnea risk. Our review shows that sleep apnea, habitual snoring, short sleep duration and poor sleep quality are important concerns for pregnant women with high body mass index. Developing screening and targeted interventions is recommended to promote efficacious perinatal care.

Evaluating the effectiveness of interprofessional advanced cardiac life support training: a sequential mixed-methods approach.

Interprofessional advanced cardiac life support (IP-ACLS) training is a holistic learning experience; thus, our research team incorporated this training into an undergraduate programme. Considering that IP-ACLS training is a new paradigm in nursing education, evaluating its effectiveness is essential. This research aimed to (1) evaluate the effectiveness of this training on improving the perceived level of interprofessional collaboration, self-efficacy and emotion regulation and (2) explore the learning experience of nursing students during training. The study design adopted a sequential mixed-method approach comprising a two-group pretest and posttest design amongst 120 students, followed by nine focus group discussions. Quantitative results demonstrated significant improvements in the perceived level of interprofessional collaboration, self-efficacy and emotion regulation at post-intervention and follow-up. Qualitative data were collected through video recording and field notes. Thematic analysis was performed following the method of Braun and Clarke. Qualitative analysis of focus group transcripts identified three themes: synergistic partnership, clinical readiness and improving further training. Quantitative and qualitative results were integrated in accordance with the mixed data analysis framework. These results complemented one another. The training provided an authentic learning experience and a good steppingstone to nursing students who are preparing to work interprofessionally in the future.

Personalised eHealth interventions in adults with overweight and obesity: A systematic review and meta-analysis of randomised controlled trials.

Given that many existing electronic health (eHealth) interventions with a general approach have limited effects, a personalised approach is necessary. We aimed to evaluate the effectiveness of personalised eHealth interventions in reducing body weight and identify the effective key features of such interventions. We searched seven databases for randomised controlled trials (RCTs) from inception until September 6, 2018. Of the 26,733 records identified, 15 RCTs were included. Meta-analysis revealed a significant reduction (-2.77 kg, 95% confidence interval - 3.54 to -2.00 kg) in the personalised eHealth intervention group compared with that in the control group (Z = -7.04, p < .001). The duration of the interventions ranged from 14 weeks and three trials had follow-up assessments at 6 and 12 months. Our subgroup analyses highlighted several crucial design elements of future personalised eHealth interventions by utilising a combination of tailored content and customised feedback with human feedback, usage of theoretical basis, short message service, device, reminder, self-monitoring, goal setting and synchronous communication for 12 to 14 weeks. Egger's regression asymmetry test suggested no evidence of publication bias (p = .458). Using meta-regression we found evidence that a statistically significant impact of age and year of publication on the effectiveness of intervention. The overall evidence grade of outcomes ranged from very low to low, hence future trials should use well-designed RCTs.

Gingival Mesenchymal Stem Cell (GMSC) Delivery System Based on RGD-Coupled Alginate Hydrogel with Antimicrobial Properties: A Novel Treatment Modality for Peri-Implantitis.

PURPOSE: Peri-implantitis is one of the most common inflammatory complications in dental implantology. Similar to periodontitis, in peri-implantitis, destructive inflammatory changes take place in the tissues surrounding a dental implant. Bacterial flora at the failing implant sites resemble the pathogens in periodontal disease and consist of Gram-negative anaerobic bacteria including Aggregatibacter actinomycetemcomitans (Aa). Here we demonstrate the effectiveness of a silver lactate (SL)-containing RGD-coupled alginate hydrogel scaffold as a promising stem cell delivery vehicle with antimicrobial properties. MATERIALS AND METHODS: Gingival mesenchymal stem cells (GMSCs) or human bone marrow mesenchymal stem cells (hBMMSCs) were encapsulated in SL-loaded alginate hydrogel microspheres. Stem cell viability, proliferation, and osteo-differentiation capacity were analyzed. RESULTS: Our results showed that SL exhibited antimicrobial properties against Aa in a dose-dependent manner, with 0.50 mg/ml showing the greatest antimicrobial properties while still maintaining cell viability. At this concentration, SL-containing alginate hydrogel was able to inhibit Aa growth on the surface of Ti discs and significantly reduce the bacterial load in Aa suspensions. Silver ions were effectively released from the SL-loaded alginate microspheres for up to 2 weeks. Osteogenic differentiation of GMSCs and hBMMSCs encapsulated in the SL-loaded alginate microspheres were confirmed by the intense mineral matrix deposition and high expression of osteogenesis-related genes. CONCLUSION: Taken together, our findings confirm that GMSCs encapsulated in RGD-modified alginate hydrogel containing SL show promise for bone tissue engineering with antimicrobial properties against Aa bacteria in vitro.

Technology-delivered personalized nutrition intervention on dietary outcomes among adults with overweight and obesity: A systematic review, meta-analysis, and meta-regression.

The prevalence of overweight and obesity has continued to increase globally, and one-size-fits-all dietary recommendations may not be suitable for different individual characteristics. A personalized nutrition intervention may be a potential solution. This review aims to evaluate the effects of the technology-delivered personalized nutrition intervention on energy, fat, vegetable, and fruit intakes among adults with overweight and obesity. A three-step comprehensive search strategy was performed from 10 databases and seven clinical registries in published and unpublished trials. A total of 46 randomized controlled trials (RCTs) involving 19,670 adults with overweight and obesity from 14 countries are included. Subgroup and meta-regression analyses were conducted. Meta-analyses showed a reduction of energy intake (-128.05, 95% CI: -197.08, -59.01) and fat intake (-1.81% energy/days, 95% CI: -3.38, -0.24, and -0.19 scores, 95% CI: -0.40, 0.02) in the intervention compared with the comparator. Significant improvements in vegetable and fruit intakes with 0.12-0.15 servings/day were observed in the intervention. Combined one- and two-way interactions had a greater effect on energy intake reduction compared with their counterparts. Meta-regression analyses revealed that no significant covariates were found. Given that the certainty of the evidence was rated as low or very low, further well-designed RCTs with long-term follow-up are warranted.

Humanoid robot-assisted interventions among children with diabetes: A systematic scoping review.

BACKGROUND: Although the humanoid robot is highly engaging for children, whether humanoid robot-assisted interventions could help in diabetes management is still unclear. OBJECTIVES: This review aimed to elucidate the roles of humanoid robot in managing diabetes and observe the effects of humanoid robot-assisted interventions among children with type I diabetes mellitus. METHODS: Arksey and O'Malley's methodological framework was used to conduct this scoping review. DATA RESOURCE: We conducted a comprehensive search in 10 databases, including IEEE Xplore Digital Library, CINAHL, Cochrane Library, EMBASE, PubMed, Scopus, Web of Science, ProQuest, WanFang, and CNKI, from May 1, 1989 to February 1, 2020. The reference lists of key texts were examined and citation chaining was conducted. In addition, our team searched gray literature and relevant journals to maximise the searching. REVIEW METHODS: Two independent reviewers examined all full articles for fitness with the eligibility criteria. Items of data extraction included the characteristics of articles, description of intervention, and mechanism of humanoid robot in improving the management of children with diabetes. RESULTS: A total of 2,755 articles were retrieved, 22 of which were included from five countries across different ethnicities. Five roles of humanoid robots in managing diabetes among children with type I diabetes mellitus were identified: disease manager, educator, advocator, emotional supporter, and communicator. To some extent, effects seem to be related to the positive improvement of affective, experiential, relational, cognitive, and behavioral perspectives from humanoid robot-assisted intervention. CONCLUSIONS: The roles of humanoids have been designed to perform their multidomain functionality. Although many positive effects were observed, the majority of selected articles have methodological issues. Future studies need an improved methodology for large-scale controlled trials.

Global Regulatory DNA Potentiation by SMARCA4 Propagates to Selective Gene Expression Programs via Domain-Level Remodeling.

The human genome encodes millions of regulatory elements, of which only a small fraction are active within a given cell type. Little is known about the global impact of chromatin remodelers on regulatory DNA landscapes and how this translates to gene expression. We use precision genome engineering to reawaken homozygously inactivated SMARCA4, a central ATPase of the human SWI/SNF chromatin remodeling complex, in lung adenocarcinoma cells. Here, we combine DNase I hypersensitivity, histone modification, and transcriptional profiling to show that SMARCA4 dramatically increases both the number and magnitude of accessible chromatin sites genome-wide, chiefly by unmasking sites of low regulatory factor occupancy. By contrast, transcriptional changes are concentrated within well-demarcated remodeling domains wherein expression of specific genes is gated by both distal element activation and promoter chromatin configuration. Our results provide a perspective on how global chromatin remodeling activity is translated to gene expression via regulatory DNA.

Corrigendum: Positively selected enhancer elements endow osteosarcoma cells with metastatic competence.

This corrects the article DOI: 10.1038/nm.4475.

Positively selected enhancer elements endow osteosarcoma cells with metastatic competence.

Metastasis results from a complex set of traits acquired by tumor cells, distinct from those necessary for tumorigenesis. Here, we investigate the contribution of enhancer elements to the metastatic phenotype of osteosarcoma. Through epigenomic profiling, we identify substantial differences in enhancer activity between primary and metastatic human tumors and between near isogenic pairs of highly lung metastatic and nonmetastatic osteosarcoma cell lines. We term these regions metastatic variant enhancer loci (Met-VELs). Met-VELs drive coordinated waves of gene expression during metastatic colonization of the lung. Met-VELs cluster nonrandomly in the genome, indicating that activity of these enhancers and expression of their associated gene targets are positively selected. As evidence of this causal association, osteosarcoma lung metastasis is inhibited by global interruptions of Met-VEL-associated gene expression via pharmacologic BET inhibition, by knockdown of AP-1 transcription factors that occupy Met-VELs, and by knockdown or functional inhibition of individual genes activated by Met-VELs, such as that encoding coagulation factor III/tissue factor (F3). We further show that genetic deletion of a single Met-VEL at the F3 locus blocks metastatic cell outgrowth in the lung. These findings indicate that Met-VELs and the genes they regulate play a functional role in metastasis and may be suitable targets for antimetastatic therapies.

Pictorial essay: imaging of peripheral nerve sheath tumours.

Peripheral nerve sheath tumours (PNST) may be benign or malignant. Benign PNSTs include neurofibroma and schwannoma. Neurogenic tumours share certain characteristic imaging features, suggested by a fusiform-shaped mass with tapered ends, the "split-fat" sign, atrophy of the muscles supplied by the involved nerve, the "fascicular sign," and the "target sign"; these imaging features are best demonstrated on magnetic resonance imaging. This pictorial essay emphasizes the characteristic signs and distinguishing features of PNSTs on imaging.

Magnetic resonance imaging of cruciate ligament injuries of the knee.

The anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) are important stabilizers of the knee. These ligaments are commonly injured in sports and motor vehicle accidents. An accurate diagnosis of cruciate ligament injuries is vital in patient care. Magnetic resonance imaging (MRI) can diagnose cruciate ligament injuries with high accuracy. The imaging diagnosis should be based on primary signs, although the secondary signs are sometimes useful, particularly in chronic cases. The detection of associated injuries of other structures of the knee, including menisci, collateral ligaments, cartilage, and bone, are also important. Accurate interpretation of the MRI examination requires a meticulous MRI technique, knowledge of the imaging anatomy, and understanding of the lesion appearance. This pictorial essay reviews the MRI appearance of normal and injured cruciate ligaments. Mechanisms of injury, primary and secondary MRI signs, and associated abnormalities are discussed.