Reassessing the association: Evaluation of a polyalanine deletion variant of RUNX2 in non-syndromic sagittal and metopic craniosynostosis.

The RUNT-related transcription factor RUNX2 plays a critical role in osteoblast differentiation, and alterations to gene dosage cause distinct craniofacial anomalies. Uniquely amongst the RUNT-related family, vertebrate RUNX2 encodes a polyglutamine/polyalanine repeat (Gln23-Glu-Ala17 in humans), with the length of the polyalanine component completely conserved in great apes. Surprisingly, a frequent 6-amino acid deletion polymorphism, p.(Ala84_Ala89)del, occurs in humans (termed 11A allele), and a previous association study (Cuellar et al. Bone 137:115395;2020) reported that the 11A variant was significantly more frequent in non-syndromic sagittal craniosynostosis (nsSag; allele frequency [AF] = 0.156; 95% confidence interval [CI] 0.126-0.189) compared to non-syndromic metopic craniosynostosis (nsMet; AF = 0.068; 95% CI 0.045-0.098). However, the gnomAD v.2.1.1 control population used by Cuellar et al. did not display Hardy-Weinberg equilibrium, hampering interpretation. To re-examine this association, we genotyped the RUNX2 11A polymorphism in 225 individuals with sporadic nsSag as parent-child trios and 164 singletons with sporadic nsMet, restricting our analysis to individuals of European ancestry. We compared observed allele frequencies to the non-transmitted alleles in the parent-child trios, and to the genome sequencing data from gnomAD v.4, which display Hardy-Weinberg equilibrium. Observed AFs (and 95% CI) were 0.076 (0.053-0.104) in nsSag and 0.082 (0.055-0.118) in nsMet, compared with 0.062 (0.042-0.089) in non-transmitted parental alleles and 0.065 (0.063-0.067) in gnomAD v.4.0.0 non-Finnish European control genomes. In summary, we observed a non-significant excess, compared to gnomAD data, of 11A alleles in both nsSag (relative risk 1.18, 95% CI 0.83-1.67) and nsMet (relative risk 1.29, 95% CI 0.87-1.92), but we did not replicate the much higher excess of RUNX2 11A alleles in nsSag previously reported (p = 0.0001).

Speech and Language Development, Hearing, and Feeding in Patients With Genetically Confirmed Crouzon Syndrome With Acanthosis Nigricans: A 36-Year Longitudinal Retrospective Review of Patients at the Oxford Craniofacial Unit.

OBJECTIVE: Crouzon syndrome with acanthosis nigricans (CAN) is caused by the specific mutation c.1172C>A (p.Ala391Glu) in the fibroblast growth factor receptor 3 gene, and has an estimated prevalence of 1:1,000,000 births. Most cases occur de novo; however, autosomal dominant inheritance may occur. The clinical presentation typically includes craniosynostosis, midface and maxillary hypoplasia, choanal atresia/stenosis, hydrocephalus, and intracranial hypertension. Patients develop acanthosis nigricans, a hyperkeratotic skin disorder. The authors present the first known study to investigate the speech, language, hearing, and feeding of patients with CAN. METHODS: A retrospective case-note review of patients with a genetically confirmed diagnosis of CAN attending the Oxford Craniofacial Unit during a 36-year period (1987-2023) was undertaken. RESULTS: Participants were 6 patients with genetically-confirmed CAN (5 females, 1 male), all cases arose de novo. All patients had craniosynostosis (n = 5/6 multisuture synostosis, n = 1/6 left unicoronal synostosis). Hydrocephalus was managed through ventriculoperitoneal shunt in 67% (n = 4/6) of patients, and 67% (n = 4/6) had a Chiari 1 malformation. Patients had a complex, multifactorial feeding history complicated by choanal atresia/stenosis (100%; n = 6/6), and significant midface hypoplasia. All patients required airway management through tracheostomy (83%; n = 5/6); and/or continuous positive airway pressure (67%; n = 4/6). All patients underwent adenotonsillectomy (100%; n = 6/6). Initial failure to thrive, low weight, and/or height were seen in 100% (n = 6/6) patients; 80% (n = 4/5) had reflux; 100% (n = 6/6) had nasogastric, or percutaneous endoscopic gastrostomy based feeding during their treatment journey. All patients had hearing loss (100%; n = 6/6). Early communication difficulties were common: receptive language disorder (50%; n = 3/6); expressive language disorder (50%; n = 3/6); and speech sound disorder in 50% (n = 3/6)-necessitating the use of Makaton in 80% of patients (n = 3/5). CONCLUSIONS: Patients with CAN experience significant respiratory, neurological, and structural obstacles to hearing, speech, language, and feeding. The authors present a recommended pathway for management to support patients in these domains.

Neoforehead Remodelling Techniques for Fronto-Orbital Advancement and Remodelling in the Oxford Craniofacial Unit: Lateral Remodelling and a Novel Central Self-Stabilizing S-Osteotomy Technique.

Fronto-orbital advancement and remodelling (FOAR) has undergone many modifications over the years, aimed at improving outcomes and reducing risks for patients. This work describes 2 techniques for remodelling the neoforehead used by the Oxford Craniofacial Unit since 1995: lateral remodelling and a central S-Osteotomy. Both methods adopt bone from the vertex as a neoforehead, but they differ in their techniques to adapt its shape to that of the newly remodelled orbital bandeau. The novel S-Osteotomy technique can be successfully applied to all FOAR procedures, irrespective of underlying synostosis and calvarial symmetry. It was originally developed for when 2 separate bony panels were required to create a neoforehead in asymmetrical cases, but was adopted for single panel neoforehead designs in metopic synostosis with the idea it may reduce temporal hollowing. An investigation of temporal hollowing in these patients who underwent either of the described methods was undertaken to assess this hypothesis with no statistically significant difference seen ( P =0.1111). Both techniques on average resulted in minimal hollowing that was not felt to require any revision, supporting the belief that temporal hollowing is a multifactorial issue. This work describes 2 successful methods of neoforehead remodelling and introduces the S-Osteotomy technique that can be applied in all FOAR procedures.

Streamlining the Diagnosis of Atypical Facial Palsies: A 5-year Review of 805 Patients.

BACKGROUND: Currently, there are no definitive guidelines in the investigation and management of atypical facial palsies (AFPs). Our aim was to determine the etiology of AFPs presenting to a tertiary facial palsy center and to review the current spectrum of diagnostic and management approaches to these conditions. METHODS: A retrospective cohort analysis of attendees at the Queen Victoria Hospital multidisciplinary facial palsy clinic over a 5-year period from 2016 to 2020 was conducted. Demographic data were collated from the QVH Research and Governance team. Those presenting with classic Bell's palsy or Ramsay-Hunt syndrome were excluded. Anyone with atypical presentations (including multiple recurrences, focal neurological deficits, polycranial neuropathies, autoimmune conditions, hemifacial spasms, hearing/balance issues, weight loss, segmental facial palsies, and gradual onset presentations) were included under the AFP category. These patients were subjected to standard serological and radiological investigations and their follow-ups were reviewed. RESULTS: A total of 849 patients were identified, and 805 had actual facial palsy presentations. Of these, 172 patients had AFP. The majority of these patients had MRI imaging tests, which were useful, but the remaining serological tests were found to correlate more with symptom clusters and specific questions rather than with random tests for all AFPs. CONCLUSIONS: Although serological and radiological investigations help in the diagnosis of AFP, specific questions and presentations help streamline the diagnosis, without affecting its accuracy whilst reducing unnecessary tests and, thereby, cost and time. We present an algorithm organized by specific questions of presentations in those with AFPs.

An Unusual Pattern of Closed Flexor Tendon Avulsion.

Closed avulsion of both flexor tendons is an uncommon injury pattern. We discuss a classic rugby jersey injury that resulted in avulsion of both flexor tendons with the flexor digitorum superficialis (FDS) avulsion incorporating a large fracture of the middle phalanx. To our knowledge, this pattern has been described only once in the literature. We propose a modification to the flexor tendon avulsion classification allowing incorporation of this injury pattern to help guide its management.

Schwann cell-like differentiated adipose stem cells promote neurite outgrowth via secreted exosomes and RNA transfer.

BACKGROUND: Adipose derived stem cells can be stimulated to produce a growth factor rich secretome which enhances axon regeneration. In this study we investigated the importance of exosomes, extracellular vesicles released by many different cell types, including stem cells and endogenous nervous system Schwann cells (SCs), on neurite outgrowth. METHODS: Adipose derived stem cells were differentiated towards a Schwann cell-like phenotype (dADSCs) by in vitro stimulation with a mix of factors (basic fibroblast growth factor, platelet derived growth factor-AA, neuregulin-1 and forskolin). Using a precipitation and low-speed centrifugation protocol the extracellular vesicles were isolated from the medium of the stem cells cultures and also from primary SCs. The conditioned media or concentrated vesicles were applied to neurons in vitro and computerised image analysis was used to assess neurite outgrowth. Total RNA was purified from the extracellular vesicles and investigated using qRT-PCR. RESULTS: Application of exosomes derived from SCs significantly enhanced in vitro neurite outgrowth and this was replicated by the exosomes from dADSCs. qRT-PCR demonstrated that the exosomes contained mRNAs and miRNAs known to play a role in nerve regeneration and these molecules were up-regulated by the Schwann cell differentiation protocol. Transfer of fluorescently tagged exosomal RNA to neurons was detected and destruction of the RNA by UV-irradiation significantly reduced the dADSCs exosome effects on neurite outgrowth. In contrast, this process had no significant effect on the SCs-derived exosomes. CONCLUSIONS: In summary, this work suggests that stem cell-derived exosomes might be a useful adjunct to other novel therapeutic interventions in nerve repair.

Postmastectomy Breast Reconstruction With the Totally Autologous Latissimus Dorsi Flap in the Thin, Small-Breasted Woman: Give It More Thought!

Introduction: Thin women have fewer autologous tissue breast reconstructive options than their higher body mass index counterparts-due to a lack of adequate donor sites. They are therefore usually offered expander/implant techniques. The total autologous latissimus dorsi flap is generally used in "well-padded" individuals, as they have enough fat on their back on which a completely autologous reconstruction could be based. When implant-based reconstruction is contraindicated (for instance due to planned adjuvant radiotherapy) or unacceptable to the patient, the total autologous latissimus dorsi flap can provide adequate tissue volume by utilizing the additional back fat deposits even in the thin, small-breasted patient. This option is often overlooked by many surgeons. Our case series assesses indications and patient and surgeon satisfaction with the cosmetic outcome of this technique. Methods: The oncological and clinical details of 6 patients with breast cancer who underwent total autologous latissimus dorsi myocutaneous flap immediate breast reconstruction by a single surgeon over an 8-year period were reviewed. An objective assessment of satisfaction with the cosmetic result was made by whether any additional surgical interventions (ipsilateral fat grafting/implant augmentation or contralateral liposuction/ reduction) were needed or not. A subjective assessment of breast symmetry by the surgeon using photographic records was also undertaken. The aesthetic outcomes were also objectively quantified using the BCCT.core software, initially developed for assessing the results of breast conservation surgery. Results: All 6 patients had small breasts and a low or normal body mass index. The mastectomies were performed for invasive carcinoma (n = 3) and extensive high-grade ductal carcinoma in situ (n = 3). Four had axillary surgery (2 sentinel lymph node biopsies and 2 axillary clearances), and 3 received adjuvant radiotherapy. All were happy with their reconstructive outcomes, and none suffered major postoperative complications or disease recurrence. None requested or needed any subsequent ipsilateral adjustment or contralateral symmetrizing procedures. Subjectively, the reconstructions provided acceptable or excellent cosmetic results. The cosmetic results were categorized as excellent or good on the BCCT.core scoring system. Conclusion: This underutilized method of totally autologous breast reconstruction in thinner patients with lower body mass indexes yielded good, well-accepted cosmetic results without recourse to adjustment procedures, contralateral balancing surgery, or complex microvascular surgery. We recommend that the total autologous latissimus dorsi flap should be given more consideration when planning immediate breast reconstruction in this challenging group of thin, small-breasted patients.

Outpatient workload in the NHS: A new challenge for the burn services?

The aim of this study was to evaluate the impact of outpatient care on activities at the Adults' Regional Burns Centre and the Children's Regional Burns Unit, Mid Yorkshire Hospitals NHS Trust UK, where outpatient and inpatient responsibilities are shared between the nursing staff. Data for all inpatient and outpatient interactions (all care related activities with the presence of a Registered Nurse (RN)) was collected prospectively by the attending RN for two consecutive months (October and November 2014). We also retrospectively collected data related to daily RN staffing levels, and Centre/Unit admissions and discharges. We compared our results with the staffing standard recommended by the National Burn Care Review 2001 and the National Burn Care Standards 2013. In the two months analysed we recorded a total number of 1232 interactions: 240 for the adult inpatient service, 385 for the adult outpatient service, 155 for the paediatric inpatient service, 452 for the paediatric outpatient service. Considering this burden and collating it with daily RN staffing levels, we highlighted an understaffing in 42.6% (26/61) of the days for the Adult Regional Burn Centre and in 100% (61/61) of the days and nights for the Children Regional Burn Unit. The impact of outpatients on the activities of a Burns Centre/Unit is a factor that should be taken into account when evaluating staffing needs and planning services. This is a variable not currently considered in any guideline for safe staffing.

The role of exosomes in peripheral nerve regeneration.

Peripheral nerve injuries remain problematic to treat, with poor functional recovery commonly observed. Injuries resulting in a nerve gap create specific difficulties for axonal regeneration. Approaches to address these difficulties include autologous nerve grafts (which are currently the gold standard treatment) and synthetic conduits, with the latter option being able to be impregnated with Schwann cells or stem cells which provide an appropriate micro-environment for neuronal regeneration to occur. Transplanting stem cells, however, infers additional risk of malignant transformation as well as manufacturing difficulties and ethical concerns, and the use of autologous nerve grafts and Schwann cells requires the sacrifice of a functioning nerve. A new approach utilizing exosomes, secreted extracellular vesicles, could avoid these complications. In this review, we summarize the current literature on exosomes, and suggest how they could help to improve axonal regeneration following peripheral nerve injury.