Unbiased assessment of genome integrity and purging of adverse outcomes at the target locus upon editing of CD4 T-cells for the treatment of Hyper IgM1.

Hyper IgM1 is an X-linked combined immunodeficiency caused by CD40LG mutations, potentially treatable with CD4 T-cell gene editing with Cas9 and a "one-size-fits-most" corrective template. Contrary to established gene therapies, there is limited data on the genomic alterations following long-range gene editing, and no consensus on the relevant assays. We developed drop-off digital PCR assays for unbiased detection of large on-target deletions and found them at high frequency upon editing.

Scalable GMP-compliant gene correction of CD4+ T cells with IDLV template functionally validated and .

Hyper-IgM1 is a rare X-linked combined immunodeficiency caused by mutations in the CD40 ligand () gene with a median survival of 25 years, potentially treatable with CD4+ T cell gene editing with Cas9 and a one-size-fits-most corrective donor template. Here, starting from our research-grade editing protocol, we pursued the development of a good manufacturing practice (GMP)-compliant, scalable process that allows for correction, selection and expansion of edited cells, using an integrase defective lentiviral vector as donor template. After systematic optimization of reagents and conditions we proved maintenance of stem and central memory phenotypes and expression and function of in edited healthy donor and patient cells recapitulating the physiological regulation.

Instrucciones de Alta por Video: Effectiveness of Video Discharge Instructions for Spanish-Speaking Caregivers in the Pediatric Emergency Department.

Introduction: Although evidence supports the addition of video discharge instructions to improve caregiver knowledge among English-speaking caregivers of children in the pediatric emergency department, there is no evidence about the effectiveness of videos for Spanish-speaking caregivers. The purpose of this study was to test whether Spanish video discharge instructions added to standard written and oral discharge instructions would result in improved knowledge and satisfaction among caregivers compared with written and oral instructions alone.

Methods: Spanish videos were created for fever, gastroenteritis, and bronchiolitis.

Evidence-Based Practice: Video-Discharge Instructions in the Pediatric Emergency Department.

Problem: While a high quality discharge from a Pediatric Emergency Department helps caregivers feel informed and prepared to care for their sick child at home, poor adherence to discharge instructions leads to unnecessary return visits, negative health outcomes, and decreased patient satisfaction. Nurses at the Inova Loudoun Pediatric ED utilized the Johns Hopkins Model of Evidence Based Practice to answer the following question: Among caregivers who have children discharged from the ED, does the addition of video discharge instructions (VDI) to standard written/verbal discharge instructions (SDI) result in improved knowledge about the child's diagnosis, treatment, illness duration, and when to seek further medical care?

Methods: A multidisciplinary team reviewed available evidence and created VDI for three common pediatric diagnoses: gastroenteritis, bronchiolitis, and fever. Knowledge assessments were collected before and after delivery of discharge instructions to caregivers for both the SDI and VDI groups.

Characterization of human mitochondrial ferritin promoter: identification of transcription factors and evidences of epigenetic control.

Mitochondrial ferritin (FtMt) is an iron storage protein belonging to the ferritin family but, unlike the cytosolic ferritin, it has an iron-unrelated restricted tissue expression. FtMt appears to be preferentially expressed in cell types characterized by high metabolic activity and oxygen consumption, suggesting a role in protecting mitochondria from iron-dependent oxidative damage. The human gene (FTMT) is intronless and its promoter region has not been described yet.