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Genome editing in human cells with targeted nucleases now enables diverse experimental and therapeutic genome engineering applications, but extension to primary human B cells remains limited. Here we report a method for targeted genetic engineering in primary human B cells, utilizing electroporation of CRISPR-Cas9 ribonucleoproteins (RNPs) to introduce gene knockout mutations at protein-coding loci with high efficiencies that in some cases exceeded 80%. Further, we demonstrate knock-in editing of targeted nucleotides with efficiency exceeding 10% through co-delivery of oligonucleotide templates for homology directed repair. We delivered Cas9 RNPs in two distinct in vitro culture systems to achieve editing in both undifferentiated B cells and activated B cells undergoing differentiation, reflecting utility in diverse experimental conditions. In summary, we demonstrate a powerful and scalable research tool for functional genetic studies of human B cell biology that may have further applications in engineered B cell therapeutics.

ObjectivesTo determine the prevalence of sleep-disordered breathing (SDB) symptoms among children with head and neck vascular malformations and to compare obstructive sleep apnea (OSA)-18 scores between children with head and neck vascular malformations and children with non-head and neck vascular malformations.Study designRetrospective cohort and prospective cross-sectional studies METHODS: Forty-three pediatric subjects with head and neck vascular malformations evaluated at a tertiary-care multidisciplinary vascular anomalies center were included in a retrospective cohort study. Eighty-three consecutive pediatric subjects with vascular malformations evaluated at the same center were included in the prospective cross-sectional study.ResultsIn the retrospective cohort study, 20 (47%) subjects with head and neck malformations had documented SDB symptoms. Of those with SDB symptoms, five (25%) required long-term tracheotomy. The children with SDB symptoms had greater vascular malformation size, more extensive pharyngeal involvement, greater vascular malformation mass effect on airway, and closer proximity of malformation to airway when compared to children without SDB symptoms. For the prospective cross-sectional study, 23% of pediatric subjects had malformations of the head and neck. Those with head and neck malformations had a higher OSA-18 score and a lower overall quality of life (QOL) score when compared to subjects with non-head and neck malformations.ConclusionNearly half of children with head and neck vascular malformations have SDB symptoms. Children with head and neck vascular malformations have a higher OSA-18 score and lower overall QOL score when compared to children with non-head and neck vascular malformations.Level of evidence4. Laryngoscope, 127:2159-2164, 2017.

Purpose of reviewChronic sialadenitis can affect patients of all age ranges and typically presents as recurrent or chronic painful swelling of the salivary glands. In children, the most common cause of sialadenitis is juvenile recurrent parotitis. Salivary stones, or sialolithiasis, are a much less common cause. Historically, for patients with chronic sialadenitis who failed conservative management, salivary gland removal was the standard treatment option. Recently, however, sialendoscopy has emerged as an effective gland-preserving procedure for sialadenitis evaluation and treatment in adults and children. The aim of this review is to discuss pediatric sialadenitis evaluation and treatment, including sialendoscopy indications, technique, and outcomes.Recent findingsSialendoscopy is a well tolerated and effective treatment for sialadenitis Sialendoscopy and salivary duct irrigation have been shown to improve frequency and severity of sialadenitis episodes in patients with juvenile recurrent parotitis. Salivary stones are managed successfully with endoscopic and combined endoscopic-assisted open approaches. Minimally invasive approaches with sialendoscopy have improved the ability to preserve salivary glands in patients with recurrent sialadenitis.SummarySialendoscopy is a well tolerated and effective procedure for recurrent sialadenitis in children.

A 3-year-old girl presented for routine closure of her tracheostomy site. She was intubated easily for the procedure, and the wound was closed with a drain in place. In recovery, the mother noticed fullness in the patient's submandibular region, and on examination, the girl had subcutaneous emphysema in the neck bilaterally. She returned to the operating room for exploration, and air was released from the surgical site. The wound was again closed with a drain in place, and the patient was extubated uneventfully. After arriving to the pediatric intensive care unit for monitoring, the patient acutely developed respiratory distress and was found to have pneumomediastinum and pneumothorax and was emergently intubated. She was observed closely, and the following day, the pneumothorax improved, and she successfully extubated without further complication.

Objectives/hypothesisThis study aimed to characterize the common salivary gland disorders presenting in the pediatric population and to describe the diagnostic and therapeutic outcomes of sialendoscopy in this population.Study designRetrospective observational study in a tertiary care pediatric medical center.MethodsMedical records review of patients under 18 years of age who presented to pediatric otolaryngology with symptoms related to a salivary gland disorder from 2002 to 2014.ResultsFifty patients were identified with an average age of 7.5 years at presentation. Eighty percent (40/50) of cases were diagnosed with juvenile recurrent parotitis (JRP), and 15 underwent sialendoscopy. The other 10 (20%) patients presented with sialolithiasis. These patients had a higher average age at presentation (12.4 vs. 6.3 years), and the majority were successfully removed with sialendoscopic techniques. Juvenile recurrent parotitis patients who underwent sialendoscopy had significantly higher costs of care during the period of observation compared to those who did not have a procedure, without a statistically significant difference in outcomes.ConclusionsSialendoscopy is an effective tool for stone retrieval in pediatric sialolithiasis. Juvenile recurrent parotitis patients who underwent sialendoscopy had outcomes similar to those selected for conservative therapy, calling into question whether the substantially higher care costs can be justified. Further prospective studies addressing cost-effectiveness will help define the role of sialendoscopy in JRP.Level of evidence4.

Head and neck lymphatic malformations can create airway management challenges requiring tracheotomy. Sirolimus, an inhibitor of mammalian target of rapamycin (mTOR), may inhibit growth of lymphatic malformations. We describe two patients born with large lymphatic malformations with improved airway symptoms following sirolimus therapy. Patient #1 underwent tracheotomy and multi-modal therapy including sirolimus with reduction in airway involvement but regrowth after discontinuation of sirolimus. Patient #2 also experienced a significant response to sirolimus allowing for extubation and discharge without tracheotomy. Early initiation of sirolimus therapy should be considered as a means to avoid tracheotomy in complex head and neck lymphatic malformations.

BackgroundThe incidence of localized intravascular coagulopathy (LIC) in venous malformations varies with lesion size and location, as well as the presence of palpable phleboliths. The development of LIC can cause pain and hemorrhage and can progress to disseminated intravascular coagulopathy (DIC) and thromboembolic disease resulting in death in some cases. Early recognition of LIC can relieve symptoms and prevent progression to life-threatening complications.ObjectiveThe aim of this work was to identify MRI features of venous malformation associated with LIC. We hypothesized that venous malformations with larger capacitance, slower flow and less physiological compression (greater stasis) were more likely to be associated with LIC.Materials and methodsIn this HIPAA-compliant and IRB-approved study, we retrospectively reviewed clinical records and MRI for consecutive patients undergoing evaluation of venous malformations at our multidisciplinary Birthmarks and Vascular Anomalies Center between 2003 and 2013. Inclusion required consensus diagnosis of venous malformation and availability of laboratory data and MRI; patients on anticoagulation or those previously undergoing surgical or endovascular treatment were excluded. LIC was diagnosed when D-dimer exceeded 1,000 ng/mL and/or fibrinogen was less than 200 mg/dL. Two board-certified radiologists assessed the following MRI features for each lesion: morphology (spongiform vs. phlebectatic), presence of phleboliths, size, location (truncal vs. extremity), and tissue type(s) involved (subcutis, muscle, bone and viscera). Univariate logistic regression analyses were used to test associations between LIC and MRI findings, and stepwise regression was applied to assess the significance of the individual imaging predictors.ResultsSeventy patients, 37 with LIC, met inclusion criteria during the 10-year study period (age: 14.5 +/- 13.6 years [mean +/- standard deviation]; 30 male, 40 female). Both elevated D-dimer and low fibrinogen were associated with the presence of phleboliths, larger lesion sizes and visceral involvement on MRI (all P

Objective: Persistent asymptomatic cervical lymphadenopathy (PACL) is a common outpatient referral diagnosis for pediatric otolaryngologists. Historically, excisional biopsy under general anesthesia has been the gold standard for diagnosis but is associated with some risks. Current literature provides little guidance on less invasive monitoring. Our hypothesis is that the majority of children who present with PACL can be safely monitored with ultrasound and avoid the risks of excisional biopsy. Study Design: A retrospective review was performed of patients <18 years of age, referred to a tertiary care children's hospital for PACL who also underwent at least 1 neck ultrasound from 2007 to 2021. Patients with acute neck infections, congenital masses, or known rheumatologic, immunologic, or malignant conditions were excluded. A multivariate logistic regression model was used to determine patient and nodal factors associated with the decision for operative management. Setting: University of California, San Francisco Pediatric Otolaryngology Department. Results: Among the 197 patients meeting inclusion criteria, 30 (15.2%) underwent surgical biopsy. Overall, 26% underwent repeat ultrasound with a mean interval of 6.6 months, and a mean decrease in nodal size of 0.34 cm. Of the 30 surgical cases, 27 (90%) patients had benign pathology. Multivariate regression analysis revealed pain (p =.04), firmness (p <.001), and lack of a normal fatty hilum on ultrasound (p =.04) as statistically significantly correlated with decisions for surgical management. Conclusion: The majority of pediatric PACL is benign and does not require an excisional biopsy to rule out lymphoma. Serial clinical follow‐up with neck ultrasound can be used to safely monitor patients. [ABSTRACT FROM AUTHOR]

Infantile hemangiomas (IHs) are benign vascular tumors. Clinical history and physical examination are the most important factors for diagnosis, with most IHs having a typical presentation. Treatment is required for some IHs that cause significant cosmetic deformity or functional compromise. Propranolol is the first-line treatment of most IHs. Ongoing research is increasing our understanding of the pathophysiology of these tumors and should help to identify future potential therapeutic targets.

ObjectiveTo develop a consensus statement on the appropriate use of computed tomography (CT) for paranasal sinus disease.Subjects and methodsA modified Delphi method was used to refine expert opinion and reach consensus by the panel.ResultsAfter 3 full Delphi rounds, 33 items reached consensus and 16 statements were dropped because of not reaching consensus or redundancy. The statements that reached consensus were grouped into 4 categories: pediatric sinusitis, medical management, surgical planning, and complication of sinusitis or sinonasal tumor. The panel unanimously agreed with 13 of the 33 statements. In addition, at least 75% of the panel strongly agreed with 14 of 33 statements across all of the categories.ConclusionsFor children, careful consideration should be taken when performing CT imaging but is needed in the setting of treatment failures and complications, either of the pathological process itself or as a result of iatrogenic (surgical) complications. For adults, imaging is necessary in surgical planning, for treatment of medical and surgical complications, and in all aspects of the complete management of patients with sinonasal and skull base pathology.

ObjectiveTo develop a consensus statement on the appropriate use of computed tomography (CT) for paranasal sinus disease.Subjects and methodsA modified Delphi method was used to refine expert opinion and reach consensus by the panel.ResultsAfter 3 full Delphi rounds, 33 items reached consensus and 16 statements were dropped because of not reaching consensus or redundancy. The statements that reached consensus were grouped into 4 categories: pediatric sinusitis, medical management, surgical planning, and complication of sinusitis or sinonasal tumor. The panel unanimously agreed with 13 of the 33 statements. In addition, at least 75% of the panel strongly agreed with 14 of 33 statements across all of the categories.ConclusionsFor children, careful consideration should be taken when performing CT imaging but is needed in the setting of treatment failures and complications, either of the pathological process itself or as a result of iatrogenic (surgical) complications. For adults, imaging is necessary in surgical planning, for treatment of medical and surgical complications, and in all aspects of the complete management of patients with sinonasal and skull base pathology.

Objectives/hypothesisTo describe the clinical presentation and airway characteristics of infants with airway hemangiomas and concomitant PHACE syndrome and to determine the prevalence of airway hemangiomas in PHACE subjects at our institution.Study designCase series.MethodsRetrospective review including clinical presentation, airway findings, treatment measures, and outcomes.ResultsA total of 23 subjects were diagnosed with definite PHACE at our institution between September 1, 2005 and September 1, 2011. Twelve (52%) of these subjects had documented airway hemangiomas, six of whom were diagnosed and treated at our institution. All six subjects underwent direct laryngoscopy and bronchoscopy by a pediatric otolaryngologist. Five (83%) subjects had subglottic hemangioma. Three subjects (50%) had additional hemangioma within the airway located on the epiglottis, vocal folds, posterior pharyngeal wall, and tracheal wall. Five subjects (83%) were treated with propranolol, five (83%) were treated with systemic steroids, and one subject received vincristine. One subject required laser ablation of subglottic hemangioma and tracheotomy. All subjects were airway symptom free at last follow-up (average, 35 months; range, 13-76 months).ConclusionsAirway hemangiomas can be a life-threatening complication of PHACE syndrome. At our institution, 52% of all PHACE subjects were diagnosed with airway hemangiomas. Early detection of airway involvement is paramount. Given the high rates of airway hemangiomas, we recommend performing direct laryngoscopy and bronchoscopy in all PHACE patients with respiratory symptoms. We recommend having a low threshold for airway evaluation in asymptomatic PHACE patients, especially those who will not be otherwise started on propranolol.

ObjectivesTo demonstrate the feasibility of drug-induced sleep endoscopy (DISE) in the pediatric population and to examine DISE results in children with persistent sleep-disordered breathing (SDB) after tonsillectomy and adenoidectomy (T&A).DesignRetrospective case series with medical chart review.SettingTertiary pediatric medical center.PatientsThirteen pediatric subjects with persistent SDB after T&A are included in the study.InterventionDrug-induced sleep endoscopy was per-formed on all patients with documentation of all sites of persistent airway obstruction.ResultsMultilevel upper-airway obstruction was identified in the majority of patients, most commonly related to tongue base obstruction, adenoid regrowth, and/or inferior turbinate hypertrophy. There were no differences among the 4 subgroups.ConclusionsFindings from DISE suggest that multiple factors contribute to airway obstruction in persistent SDB after T&A. Further research can address the extent to which directed surgical treatment can improve outcomes in these patients.

This article discusses the management of newborn traumatic nasal deformity using a nostril retainer. The incidence of nasal septal deformity in newborns varies, and the etiology is debated among authors. The article presents three cases in which a nostril retainer was used to treat newborns with nasal deformity or obstruction. The results showed significant improvement in nasal position, and the retainer was well-tolerated by the infants. The article suggests that the nostril retainer can be a simple and non-invasive method for addressing postnatal nasal deformity and obstruction in newborns. [Extracted from the article]

Objective: To describe the prevalence and clinical characteristics of airway findings in a multi-institutional cohort of PHACE patients. Study Design: Multicenter retrospective case series. Setting: Multidisciplinary vascular anomalies clinics at 2 institutions. Methods: Data were collected from the electronic medical record, including clinical presentation, airway findings, treatment, and outcomes. Results: Of 55 PHACE patients, 22 (40%) had airway hemangiomas. Patients with airway involvement were more commonly female (P =.034, odds ratio [OR] 23, 95% confidence interval [CI] 1.3-410) and of Caucasian ethnicity (P =.020, OR 5.3, 95% CI 1.3-21). Anatomically, patients with bilateral S3 involvement had higher rates of airway disease (P =.0012, OR 15, 95% CI 2.9-77). Most patients with airway hemangiomas had stridor (68%). Of the patients managed in the propranolol era (2008 or later, n = 35), 14 had airway involvement. All 14 were treated with propranolol, whereas 13 (62%) of 21 nonairway patients were treated with propranolol. The average treatment duration was longer in the airway patients (22.1 vs 16.7 months). All patients who underwent tracheostomy (n = 4) did so before 2008. Conclusion: Risk factors for airway involvement in PHACE include female gender, Caucasian ethnicity, and stridor. Since the widespread use of propranolol, fewer patients have required surgical management of their airway disease. Given the high prevalence of airway involvement even in patients without stridor, assessment of the airway is a crucial component of a comprehensive PHACE workup. [ABSTRACT FROM AUTHOR]