Mutation in Eftud2 causes craniofacial defects in mice via mis-splicing of Mdm2 and increased P53.

EFTUD2 is mutated in patients with mandibulofacial dysostosis with microcephaly (MFDM). We generated a mutant mouse line with conditional mutation in Eftud2 and used Wnt1-Cre2 to delete it in neural crest cells. Homozygous deletion of Eftud2 causes brain and craniofacial malformations, affecting the same precursors as in MFDM patients. RNAseq analysis of embryonic heads revealed a significant increase in exon skipping and increased levels of an alternatively spliced Mdm2 transcript lacking exon 3. Exon skipping in Mdm2 was also increased in O9-1 mouse neural crest cells after siRNA knock-down of Eftud2 and in MFDM patient cells. Moreover, we found increased nuclear P53, higher expression of P53-target genes and increased cell death. Finally, overactivation of the P53 pathway in Eftud2 knockdown cells was attenuated by overexpression of non-spliced Mdm2, and craniofacial development was improved when Eftud2-mutant embryos were treated with Pifithrin-α, an inhibitor of P53. Thus, our work indicates that the P53-pathway can be targeted to prevent craniofacial abnormalities and shows a previously unknown role for alternative splicing of Mdm2 in the etiology of MFDM.

Craniofacial Defects in Embryos with Homozygous Deletion of Eftud2 in Their Neural Crest Cells Are Not Rescued by Trp53 Deletion.

Embryos with homozygous mutation of Eftud2 in their neural crest cells (Eftud2ncc-/-) have brain and craniofacial malformations, hyperactivation of the P53-pathway and die before birth. Treatment of Eftud2ncc-/- embryos with pifithrin-α, a P53-inhibitor, partly improved brain and craniofacial development. To uncover if craniofacial malformations and death were indeed due to P53 hyperactivation we generated embryos with homozygous loss of function mutations in both Eftud2 and Trp53 in the neural crest cells. We evaluated the molecular mechanism underlying craniofacial development in pifithrin-α-treated embryos and in Eftud2; Trp53 double homozygous (Eftud2ncc-/-; Trp53ncc-/-) mutant embryos. Eftud2ncc-/- embryos that were treated with pifithrin-α or homozygous mutant for Trp53 in their neural crest cells showed reduced apoptosis in their neural tube and reduced P53-target activity. Furthermore, although the number of SOX10 positive cranial neural crest cells was increased in embryonic day (E) 9.0 Eftud2ncc-/-; Trp53ncc-/- embryos compared to Eftud2ncc-/- mutants, brain and craniofacial development, and survival were not improved in double mutant embryos. Furthermore, mis-splicing of both P53-regulated transcripts, Mdm2 and Foxm1, and a P53-independent transcript, Synj2bp, was increased in the head of Eftud2ncc-/-; Trp53ncc-/- embryos. While levels of Zmat3, a P53- regulated splicing factor, was similar to those of wild-type. Altogether, our data indicate that both P53-regulated and P53-independent pathways contribute to craniofacial malformations and death of Eftud2ncc-/- embryos.

Transmembrane emp24 domain proteins in development and disease.

Regulated transport through the secretory pathway is essential for embryonic development and homeostasis. Disruptions in this process impact cell fate, differentiation and survival, often resulting in abnormalities in morphogenesis and in disease. Several congenital malformations are caused by mutations in genes coding for proteins that regulate cargo protein transport in the secretory pathway. The severity of mutant phenotypes and the unclear aetiology of transport protein-associated pathologies have motivated research on the regulation and mechanisms through which these proteins contribute to morphogenesis. This review focuses on the role of the p24/transmembrane emp24 domain (TMED) family of cargo receptors in development and disease.

Post-COVID-19 Mental Health Distress in 13 Million Youth: A Retrospective Cohort Study of Electronic Health Records.

OBJECTIVE: To investigate the impact of the SARS-CoV-2 infection on the rates of mental disorders in youth. METHOD: The study involved 7,519,465 children and 5,338,496 adolescents from the TriNetX Research Network, all without prior mental disorder histories. Among them, 290,145 children and 223,667 adolescents had SARS-CoV-2-positive tests or confirmed COVID-19 diagnoses. Kaplan-Meier survival analysis was used to evaluate the probability of developing new mental disorders (any codes in International Classification of Diseases, Tenth Revision, Clinical Modification (ICD-10-CM) F01-F99 category and suicidal behaviors) within 2 years post infection, compared to the propensity score-matched youth who were never infected. RESULTS: Within 2 years post SARS-CoV-2 infection, children had a probability of 0.15 in acquiring new psychiatric diagnoses, compared to 0.026 for matched non-infected children; adolescents had a 0.19 probability against 0.05 for their non-infected counterparts. The hazard ratio (HR) was 6.0 (95% CI = 5.8-6.3) for children and 4.2 for adolescents (95% CI = 4.1-4.4), with children vs adolescents HR of 1.4 (95% CI = 1.36-1.51). Elevated HRs were observed for almost all subcategories of mental disorders and suicidal behaviors, with variations based on sex, severity of SARS-CoV-2 infection, and viral variants. COVID-19 was similar to other respiratory infections and was associated with a similarly increased rate of mental disorders in adolescents, but had a significantly higher effect on children (HR = 1.57, 95% CI =1.53-1.61). CONCLUSION: This study revealed significant mental health distress following SARS-CoV-2 infection in youth, which was more pronounced in children than in adolescents. These findings underscore the urgent need to support at-risk youth, particularly those who contracted SARS-CoV-2 at younger ages and had more severe infections. CLINICAL GUIDANCE.

Long-Term Hand and Shoulder Function in Children following Early Surgical Intervention for a Birth-Related Upper Brachial Plexus Injury.

Purpose To better understand the long-term hand and shoulder outcomes of upper brachial plexus birth injuries. Methods We evaluated shoulder and hand function in 32 patients (13 males; 19 females) with a C5/C6 birth injury history). All patients had undergone primary nerve surgery as infants, and 12 underwent a simultaneous shoulder procedure as they presented with a fixed internal rotation contracture of the shoulder. On average, all patients were evaluated and examined 15 years postoperatively. The shoulder function was evaluated using the Miami Shoulder Scale. Hand function was measured by the 9-hole peg test (9-HPT) and statistical analysis included comparison of 9-HPT time against normative data using the Student's t -test. Results The cohort includes 22 right-hand-dominant and 10 left-hand-dominant patients. Mean age at surgery was 10 months; mean age at follow-up was 15 years ± 2 years 2 months. Cumulative shoulder function was "good" or "excellent" (Miami score) in 23 patients. For 9-HPT, 23 out of 32 patients seen had an involved hand with a significant alteration in function. Conclusion Early nerve surgery in cases of upper brachial plexus birth injuries result in the desired outcome. To ensure timely and targeted therapy for any residual deficits, it is imperative that limitations in hand function among children with an Erb's palsy.

The relationship between postpartum pain and mother-infant bonding: A prospective observational study.

INTRODUCTION: Postpartum pain is associated with impaired maternal recovery and may influence mother-infant bonding. METHODS: Participants who underwent a vaginal or cesarean delivery were assessed 24 h postpartum. Postpartum pain intensity was measured using the Verbal Numeric Score (VNS) (0-10) and classified as non-severe (<8) or severe pain (≥8). Maternal-infant bonding was evaluated using the Post-Partum Bonding Questionnaire (PBQ; 0-125), with a score > 5 defining impaired bonding. Demographic data included age, BMI, parity, education level, economic status, partnership, prior history of depression, familial history of depression, desire to breastfeed, epidural analgesia during labor, rooming in, and Edinburgh Postnatal Depression Scale (EPDS). Data were analyzed using 2 separate multivariable logistic regression models for vaginal and cesarean deliveries, where maximum postpartum pain was the independent variable and impaired postpartum bonding was the dependent variable and controlled for the other factors collected. RESULTS: Severe postpartum pain (VNS ≥ 8) showed no significant relationship with impaired bonding when controlling for confounding variables. In vaginal deliveries, there was an association between a history of depression and impaired bonding (Odds Ratio 2.2 [1.07-4.65], p = 0.04) and EPDS > 10 and impaired bonding (OR 11.5 [3.2-73.6], p < 0.001). For cesarean deliveries, rooming in with the baby had a protective effect (OR 11.5 [3.2-73.6], p < 0.001). CONCLUSIONS: Contrary to expectations, severe postpartum pain did not influence maternal-infant binding in the cohort of patients with vaginal and cesarean deliveries. Instead, factors such as maternal mental health and rooming-in practices appeared to exert more significant influence. CLINICAL TRIAL REGISTRATION: NCT05206552.