High-Resolution Whole-Genome Analysis of Sister-Chromatid Contacts.

Sister-chromatid cohesion describes the orderly association of newly replicated DNA molecules behind replication forks. It plays an essential role in the maintenance and faithful transmission of genetic information. Cohesion is created by DNA topological links and proteinaceous bridges, whose formation and deposition could be potentially affected by many processes. Current knowledge on cohesion has been mainly gained by fluorescence microscopy observation. However, the resolution limit of microscopy and the restricted number of genomic positions that can be simultaneously visualized considerably hampered progress. Here, we present a high-throughput methodology to monitor sister-chromatid contacts (Hi-SC2). Using the multi-chromosomal Vibrio cholerae bacterium as a model, we show that Hi-SC2 permits to monitor local variations in sister-chromatid cohesion at a high resolution over a whole genome.

Scandcleft randomized trials of primary surgery for unilateral cleft lip and palate: Speech proficiency at 10 years of age.

BACKGROUND & AIM: To assess consonant proficiency and velopharyngeal function in 10-year-old children born with unilateral cleft lip and palate (UCLP) within the Scandcleft project. METHODS & PROCEDURES: Three parallel group, randomized, clinical trials were undertaken as an international multicentre study by nine cleft teams in five countries. Three different surgical protocols for primary palate repair (Arm B-Lip and soft palate closure at 3-4 months, hard palate closure at 36 months, Arm C-Lip closure at 3-4 months, hard and soft palate closure at 12 months, and Arm D-Lip closure at 3-4 months combined with a single-layer closure of the hard palate using a vomer flap, soft palate closure at 12 months) were tested against a common procedure (Arm A-Lip and soft palate closure at 3-4 months followed by hard palate closure at 12 months) in the total cohort of 431 children born with a non-syndromic UCLP. Speech audio and video recordings of 399 children were available and perceptually analysed. Percentage of consonants correct (PCC) from a naming test, an overall rating of velopharyngeal competence (VPC) (VPC-Rate), and a composite measure (VPC-Sum) were reported. OUTCOMES & RESULTS: The mean levels of consonant proficiency (PCC score) in the trial arms were 86-92% and between 58% and 83% of the children had VPC (VPC-Sum). Only 50-73% of the participants had a consonant proficiency level with their peers. Girls performed better throughout. Long delay of the hard palate repair (Arm B) indicated lower PCC and simultaneous hard and soft palate closure higher (Arm C). However, the proportion of participants with primary VPC (not including velopharyngeal surgeries) was highest in Arm B (68%) and lowest in Arm C (47%). CONCLUSIONS & IMPLICATIONS: The speech outcome in terms of PCC and VPC was low across the trials. The different protocols had their pros and cons and there is no obvious evidence to recommend any of the protocols as superior. Aspects other than primary surgical method, such as time after velopharyngeal surgery, surgical experience, hearing level, language difficulties and speech therapy, need to be thoroughly reviewed for a better understanding of what has affected speech outcome at 10 years. WHAT THIS PAPER ADDS: What is already known on the subject Speech outcomes at 10 years of age in children treated for UCLP are sparse and contradictory. Previous studies have examined speech outcomes and the relationship with surgical intervention in 5-year-olds. What this study adds to the existing knowledge Speech outcomes based on standardized assessment in a large group of 10-year-old children born with UCLP and surgically treated according to different protocols are presented. While speech therapy had been provided, a large proportion of the children across treatment protocols still needed further speech therapy. What are the potential or actual clinical implications of this work? Aspects other than surgery and speech function might add to the understanding of what affects speech outcome. Effective speech therapy should be available for children in addition to primary surgical repair of the cleft and secondary surgeries if needed.

VPI Management in SATB2 Syndrome: Use of MRI to Evaluate Anatomy and Physiology in Non-Cleft VPI.

This clinical case study describes the velopharyngeal anatomy and physiology in a patient who presented with SATB2-associated syndrome (SAS) and velopharyngeal insufficiency (VPI) in the absence of an overt cleft palate. The clinical presentation, treatment, outcome, and the contribution of anatomical findings from MRI to surgical treatment planning for this rare genetic disorder, SAS, are described. This case study contributes to our current understanding of the anatomy and physiology of the velopharyngeal mechanism in an individual born with SAS and non-cleft VPI. It also details the changes following bilateral buccal myomucosal flaps in this patient.

Genetic and mutational analysis of virulence traits and their modulation in an environmental toxigenic Vibrio cholerae non-O1/non-O139 strain, VCE232.

Vibrio cholerae O1 and O139 isolates deploy cholera toxin (CT) and toxin-coregulated pilus (TCP) to cause the diarrhoeal disease cholera. The ctxAB and tcpA genes encoding CT and TCP are part of two acquired genetic elements, the CTX phage and Vibrio pathogenicity island-1 (VPI-1), respectively. ToxR and ToxT proteins are the key regulators of virulence genes of V. cholerae O1 and O139. V. cholerae isolates belonging to serogroups other than O1/O139, called non-O1/non-O139, are usually devoid of virulence-related elements and are non-pathogenic. Here, we have analysed the available whole genome sequence of an environmental toxigenic V. cholerae non-O1/non-O139 strain, VCE232, carrying the CTX phage and VPI-1. Extensive bioinformatics and phylogenetic analyses indicated high similarity of the VCE232 genome sequence with the genome of V. cholerae O1 strains, including organization of the VPI-1 locus, ctxAB, tcpA and toxT genes, and promoters. We established that the VCE232 strain produces an optimal amount of CT at 30 °C under AKI conditions. To investigate the role of ToxT and ToxR in the regulation of virulence factors, we constructed ΔtoxT, ΔtoxR and ΔtoxTΔtoxR deletion mutants of VCE232. Extensive genetic analyses of these mutants indicated that the toxT and toxR genes of VCE232 are crucial for CT and TCP production. However, unlike O1 isolates, the presence of either toxT or toxR gene is sufficient for optimal CT production in VCE232. In addition, the VCE232 ΔtoxR mutant showed differential regulation of the major outer membrane proteins, OmpT and OmpU. This is the first attempt to explore the regulation of expression of major virulence genes and regulators in an environmental toxigenic V. cholerae non-O1/non-O139 strain.

Posterior pharyngeal wall augmentation in post-adenoidectomy velopharyngeal insufficiency.

PURPOSE: To assess the efficacy of posterior pharyngeal wall augmentation using septal or conchal cartilages with other bulks-according to the persistent gap and the individual anatomy of each patient-in improving velopharyngeal function in patients who acquired persistent velopharyngeal insufficiency (VPI) post-adenoidectomy. METHODS: Observational descriptive prospective case series of 24 patients (their ages ranged between 3 and 26 years) who developed persistent VPI post-adenoidectomy (more than 3 months) although they had normal speech resonance before adenoidectomy. RESULTS: The present study demonstrated that statistically significant improvement in auditory perceptual assessment (APA) was found regarding all obligatory speech disorders and unintelligibility of speech. Significant improvement was observed in the degree of velar mobility, size of the persistent gap, and the gap distance between velum and posterior pharyngeal wall at rest and during phonation in post-operative evaluation versus pre-operative. A significant change was observed in the closure pattern of the velopharyngeal port (VPP) as all patients turned to coronal closure. CONCLUSIONS: Posterior pharyngeal wall augmentation could be used in VPI post-adenoidectomy up to 7 mm and lead to better speech outcomes. Also, it revealed that using conchal and/or septal cartilage as a graft regardless of the patient's age is a safe procedure.

Predictive Factors for Velopharyngeal Insufficiency Following Primary Cleft Palate Repair.

OBJECTIVE: Velopharyngeal insufficiency (VPI) remains a known complication of primary palatoplasty. We sought to identify factors associated with the incidence of VPI and create a predictive model for VPI development in our population. DESIGN: A single-institution, retrospective review. SETTING: Multidisciplinary clinic in a tertiary academic institution. PATIENTS: A total of 453 consecutive patients undergoing primary palatoplasty from 1999 to 2016 were reviewed. Inclusion required follow-up past age 5. Patients who were non-verbal, and thus unable to undergo speech evaluation, were excluded. MAIN OUTCOME MEASURES: Primary outcome was VPI, defined as revision palatoplasty or recommendation by speech-language pathology. RESULTS: Of 318 patients included, 179 (56%) were male. Median age at primary repair was 1.0 years (0.9-1.1) with a median age of 8.8 years at last follow-up. One hundred nineteen (37%) patients developed VPI at a median age of 5.0 years (3.8-6.5). Higher rates were seen with posterior fistula (65% vs 14%, P <.01) and straight-line repair (41% vs 9%, P <.01), with lower rates in patients with Veau I clefts (22% vs 39%, P <.05). Patients with VPI were older at last follow-up. Following multivariate regression, factors remaining significant were posterior fistula (odds ratio [OR]: 11.3, 95% CI: 6.1-22.0), primary Furlow repair (OR: 0.18, 95% CI: 0.03-0.68), genetic diagnoses (OR: 2.92, 95% CI: 1.1-7.9), and age at last follow-up (OR: 1.11, 95% CI: 1.01-1.2). CONCLUSIONS: Length of follow-up, posterior fistulae, and genetic diagnoses are associated with VPI formation. Furlow repair may protect against formation of VPI. Use of allograft, Veau class, birth type, birth weight, and race are not independently associated with VPI formation.

The value of CT radiomics features to predict visceral pleural invasion in ≤3 cm peripheral type early non-small cell lung cancer.

OBJECTIVE: To investigate predictive value of CT-based radiomics features on visceral pleural invasion (VPI) in ≤3.0 cm peripheral type early non-small cell lung cancer (NSCLC). METHODS: A total of 221 NSCLC cases were collected. Among them, 115 are VPI-positive and 106 are VPI-negative. Using a stratified random sampling method, 70% cases were assigned to training dataset (n = 155) and 30% cases (n = 66) were assigned to validation dataset. First, CT findings, imaging features, clinical data and pathological findings were retrospectively analyzed, the size, location and density characteristics of nodules and lymph node status, the relationship between lesions and pleura (RAP) were assessed, and their mean CT value and the shortest distance between lesions and pleura (DLP) were measured. Next, the minimum redundancy-maximum relevance (mRMR) and least absolute shrinkage and selection operator (LASSO) features were extracted from the imaging features. Then, CT imaging prediction model, texture feature prediction model and joint prediction model were built using multifactorial logistic regression analysis method, and the area under the ROC curve (AUC) was applied to evaluate model performance in predicting VPI. RESULTS: Mean diameter, density, fractal relationship with pleura, and presence of lymph node metastasis were all independent predictors of VPI. When applying to the validation dataset, the CT imaging model, texture feature model, and joint prediction model yielded AUC = 0.882, 0.824 and 0.894, respectively, indicating that AUC of the joint prediction model was the highest (p < 0.05). CONCLUSION: The study demonstrates that the joint prediction model containing CT morphological features and texture features enables to predict the presence of VPI in early NSCLC preoperatively at the highest level.

Agreements between speech language pathologists and naïve listeners' judgements of intelligibility in children with cleft palate.

Speech-language pathologists (SLPs) working in interdisciplinary craniofacial teams need to know how much their judgments of speech intelligibility could predict the patient's communication difficulties with non-clinical communicative partners. This study examines the relationship between experienced SLPs and naïve listeners in judgments of speech intelligibility in speakers with cleft palate. A speech perception study was conducted using speech samples from 20 speakers with velopharyngeal insufficiency (VPI) following cleft palate. Speech samples were presented to 70 naïve listeners and 10 SLPs who were experts in cleft palate treatment. Speech intelligibility scores from naïve listeners' orthographic transcriptions were obtained as the percentage of correctly identified words. Speech intelligibility scores from SLPs were obtained using a five-point rating scale. Spearman rank correlation indicated a very high level of overall agreement between naïve listeners and SLPs at the speaker level scores (rs = -.94, p < .001). While the listeners' judgment seems highly related across highly intelligible speakers, the differences in agreements increase when the speaker is unintelligible. The high correlations between scores for naïve listeners and SLPs suggest that speech intelligibility in children with VPI could be predicted by ratings done in the clinic by expert SLPs.

Simultaneous gene expression and multi-gene silencing in Zea mays using maize dwarf mosaic virus.

BACKGROUND: Maize dwarf mosaic virus (MDMV), a member of the genus Potyvirus, infects maize and is non-persistently transmitted by aphids. Several plant viruses have been developed as tools for gene expression and gene silencing in plants. The capacity of MDMV for both gene expression and gene silencing were examined. RESULTS: Infectious clones of an Ohio isolate of MDMV, MDMV OH5, were obtained, and engineered for gene expression only, and for simultaneous marker gene expression and virus-induced gene silencing (VIGS) of three endogenous maize target genes. Single gene expression in single insertion constructs and simultaneous expression of green fluorescent protein (GFP) and silencing of three maize genes in a double insertion construct was demonstrated. Constructs with GFP inserted in the N-terminus of HCPro were more stable than those with insertion at the N-terminus of CP in our study. Unexpectedly, the construct with two insertion sites also retained insertions at a higher rate than single-insertion constructs. Engineered MDMV expression and VIGS constructs were transmissible by aphids (Rhopalosiphum padi). CONCLUSIONS: These results demonstrate that MDMV-based vector can be used as a tool for simultaneous gene expression and multi-gene silencing in maize.

Impact of Surgery for Velopharyngeal Insufficiency on Eustachian Tube Function in Children: Pharyngeal Flap Versus Sphincter Pharyngoplasty.

OBJECTIVE: Velopharyngeal insufficiency (VPI) is a common speech disorder in patients with a history of cleft palate (CP) or 22q11.2 deletion syndrome. Pharyngeal flap (PF) and sphincter pharyngoplasty (SP) are 2 common surgeries to treat this disorder by decreasing unwanted nasal air emission and hypernasal resonance. Because Eustachian tube dysfunction (ETD) in patients with CP may be more frequent after surgery for VPI, we examined whether ETD was associated with either type of surgery. DESIGN: Retrospective cohort study. SETTING: Children's hospital-based tertiary referral center. PATIENTS: A total of 225 children with VPI who underwent primary PF (201) or SP (24) between 2006 and 2017. OUTCOME MEASURES: We examined differences in risk of ETD according to both surgical groups and proxies for postoperative nasal obstruction. These proxies included postoperative resonance measures and development of obstructive sleep apnea (OSA). RESULTS: Both surgical groups had similar preoperative measures, except the PF group had higher hypernasality by PSA. Postoperatively, the PF group demonstrated lower hypernasal resonance by nasometry and PSA. There were no differences between PF and SP groups with regard to ETD. Proxies for postoperative nasal obstruction also were not predictive of postoperative ETD. Degree of CP and younger age were found to be risk factors for ETD. CONCLUSION: There was no significant difference in the effects of PF and SP on ETD in this study. Neither lower hypernasality nor incidence of OSA had any impact on ETD. Degree of CP and younger age were the only significant risk factors for ETD that this study identified.

Velopharyngeal Muscle Morphology in Children With Unrepaired Submucous Cleft Palate: An Imaging Study.

OBJECTIVE: To identify quantitative and qualitative differences in the velopharyngeal musculature and surrounding structures between children with submucous cleft palate (SMCP) and velopharyngeal insufficiency (VPI) and noncleft controls with normal anatomy and normal speech. METHODS: Magnetic resonance imaging was used to evaluate the velopharyngeal mechanism in 20 children between 4 and 9 years of age; 5 with unrepaired SMCP and VPI. Quantitative and qualitative measures of the velum and levator veli palatini in participants with symptomatic SMCP were compared to noncleft controls with normal velopharyngeal anatomy and normal speech. RESULTS: Analysis of covariance revealed that children with symptomatic SMCP demonstrated increased velar genu angle (15.6°, P = .004), decreased α angle (13.2°, P = .37), and longer (5.1 mm, P = .32) and thinner (4 mm, P = .005) levator veli palatini muscles compared to noncleft controls. Qualitative comparisons revealed discontinuity of the levator muscle through the velar midline and absence of a musculus uvulae in children with symptomatic SMCP compared to noncleft controls. CONCLUSIONS: The levator veli palatini muscle is longer, thinner, and discontinuous through the velar midline, and the musculus uvulae is absent in children with SMCP and VPI compared to noncleft controls. The overall velar configuration in children with SMCP and VPI is disadvantageous for achieving adequate velopharyngeal closure necessary for nonnasal speech compared to noncleft controls. These findings add to the body of literature documenting levator muscle, musculus uvulae, and velar and craniometric parameters in children with SMCP.

The effect of velopharyngeal insufficiency on voice quality in Estonian Children with Cleft Palate.

Most children born with cleft palate (CP) or cleft lip and palate (CL/P) have velopharyngeal insufficiency (VPI), some degree of hypernasal resonance, articulation disorders and laryngeal dysphonia. Combination of different CL/P specific problems may mask laryngeal dysphonia and therefore, it may remain undiagnosed and untreated by clinicians. The research aimed to study the effect of VPI on voice quality in Estonian CL/P children. We included 18 CL/P and 79 healthy children. Combination of objective (Multi-Dimensional Voice Program (MDVP)) and subjective (Pediatric Voice Handicap Index (pVHI), GRBAS scale, video-nasoendoscopy (VNE), video-laryngostroboscopy (VLS)) assessment methods were assisted and performed by our multidisciplinary cleft teams. We found that (1) overall quality of life is greatly affected by voice, resonance and articulation disorders in CL/P group, (2) more than half of the CL/P children had morphological changes of the vocal folds, (3) the severity of VPI did not result in worse outcomes of acoustic parameters of voice.

Relationship of Velopharyngeal Insufficiency With Face Mask Therapy in Patients With Cleft Lip and Palate.

OBJECTIVE: To determine whether orthodontic/dentofacial orthopedic maxillary protraction face mask therapy induces changes in velopharyngeal functioning in a cohort of pediatric patients having cleft palate with or without cleft lip. DESIGN: Retrospective chart review. SETTING: A children's hospital in the United States. PARTICIPANTS: Forty-three pediatric patients with cleft palate, with or without cleft lip, syndromic or with isolated clefts, who received face mask therapy from January 2009 to April 2016. INTERVENTION: Clinical data were extracted for review and analysis from medical records obtained from the Cleft Database/Research Registry (CDB-RR). MAIN OUTCOME MEASURES: Pittsburgh Weighted Speech Scores (PWSS) before and after therapy. RESULTS: There was a significant increase in PWSS after face mask therapy for patients with a PWSS score of 0 prior to treatment. Patients with PWSS >0 before treatment remained largely stable after face mask therapy. Maxillary advancement was not significantly associated with change in PWSS or fistula presence/absence. CONCLUSIONS: There is an increased risk of velopharyngeal insufficiency with maxillary protraction face mask treatment in patients with cleft palate. Patient counseling and obtaining consent regarding speech changes during treatment are recommended.

Novel treatment for mild and moderate velopharyngeal insufficiency using implantable AlloDerm for posterior pharyngeal wall augmentation and review of surgical repair techniques.

PURPOSE: Mild and moderate velopharyngeal insufficiency is a relatively common structural defect of the velopharyngeal sphincter that occurs congenitally or secondarily to various medical conditions resulting in speech inadequacy. Currently, multiple surgical methods exist to treat mild and moderate velopharyngeal insufficiency; however, the revision rates are high and the outcomes are variable. This case series describes a novel technique using implantable AlloDerm to repair the posterior pharyngeal wall to treat mild and moderate velopharyngeal insufficiency. MATERIALS AND METHODS: This paper presents four patients with mild or moderate velopharyngeal insufficiency who were treated with implantable AlloDerm in the posterior pharyngeal wall at a large, safety-net hospital in New England from 2000 to 2019. Additionally, a review of surgical repair techniques for velopharyngeal insufficiency was conducted with synthesis of a qualitative overview. RESULTS: There were sufficient follow-up data in three of these patients. All three reported subjective improvements in symptoms after the procedure. One patient had implant extrusion one month following the procedure with subsequent removal. CONCLUSION: Ultimately, implantable AlloDerm for posterior pharyngeal wall augmentation is a useful, low risk method for treating mild to moderate velopharyngeal insufficiency.

Prosthetic Rehabilitation with Palatal Lift/Augmentation in a Patient with Neurologic/Motor Deficit Due To Cancer Therapy for Chondrosarcoma.

This clinical report describes the prosthetic rehabilitation of a 25-year-old man with a history of grade II chondrosarcoma at the skull base who had undergone surgical resection and thereafter developed velopharyngeal incompetency (VPI), dysarthria, and dysphagia. Upon baseline fiberoptic endoscopic evaluation of swallowing (FEES), the patient had an atypical pattern of VPI with minimal to no velar lift during speech, blow, or suck tasks, but near complete velar lift and seal during swallowing. A palatal augmentation prosthesis combined with a resilient palatal lift extension was fabricated to enhance speech by displacing the soft palate and to decrease hypernasality, while avoiding interference with bolus transport. A resilient wrought wire extension was necessary to accommodate the velar movement upon swallowing while keeping the integrity of the velar lift during speech. In conclusion, this unique combination prosthesis was able to help the patient's atypical pattern of VPI by improving speech and preserving swallowing function, which was confirmed during a post-endoscopic evaluation.

Elucidating the mechanism governing cell rotation under DEP using the volumetric polarization and integration method.

Cell rotation can be achieved by utilizing rotating electric fields through which torques are generated due to phase difference between the dipole moment of cells and the external electric field. While reports of cell rotation under non-rotating electrical fields, such as dielectrophoresis (DEP), are abound, the underlying mechanism is not fully understood. Because of this, contradicting arguments remain regarding if a single cell can rotate under conventional DEP. What's more, the current prevailing DEP theory is not adequate for identifying the cause for such disagreements. In this work we applied our recently developed Volumetric Polarization and Integration (VPI) method to investigate the possible causes for cell rotation under conventional DEP. Three-dimensional (3D) computer models dealing with a cell in a DEP environment were developed to quantify the force and torque imparted on the cell by the external DEP field using COMSOL Multiphysics software. Modeling results suggest that eccentric inclusions with low conductivity inside the cell will generate torques (either in clockwise or counter-clockwise directions) sufficient to cause cell rotation under DEP. For validation of modeling predictions, experiments with rat adipose stem cells containing large lipid droplets were conducted. Good agreement between our modeling and experimental results suggests that the VPI method is powerful in elucidating the underlying mechanisms governing the complicated DEP phenomena.

Barley stripe mosaic virus (BSMV) as a virus-induced gene silencing vector in maize seedlings.

Barley stripe mosaic virus (BSMV) was the first reported and still widely used virus-induced gene silencing (VIGS) vector for monocotyledons including wheat and barley. Despite BSMV's reported infectivity on maize (Zea mays), the use of the virus as a vector in maize has not been optimized. Here, we assayed infectivity of BSMV in different maize cultivars by vascular puncture inoculation. Through knockdown of the endogenous host phytoene desaturase gene, we demonstrate for the first time that BSMV can be used as a VIGS vector in maize. This adds BSMV to the repertoire of tools available for functional studies in maize.

The Detailed Comparison of Cell Death Detected by Annexin V-PI Counterstain Using Fluorescence Microscope, Flow Cytometry and Automated Cell Counter in Mammalian and Microalgae Cells.

The evaluation of cell wellness is an important task for molecular biology research. This mainly comprises the assessment for morphology and viability of culturing cells. Annexin V-Propidium iodide counterstaining has been currently one of the common and easy methods to discriminate apoptotic and necrotic cell profiles. The method is operated by fluorescence-based detection of counterstain via laser beam-employed instruments including flow cytometer, fluorescence microscope and automated cell counter. The detection is primarily conducted based on the same principle; however the efficiency of instruments may vary. Here we evaluated the efficiency of those instruments for the clear-cut detection of cell death through various mammalian and microalgae cell lines. To the best of our knowledge, this is the first study revealing comparative analyses of apoptotic and necrotic cells in mammalian and microalgae cells using Annexin V-PI counterstain detected by flow cytometer, fluorescence microscope and automated cell counter. Fluorescence microscope and cell counter instruments were also tested and compared for the traditional trypan blue-based cell viability detection performance. For these, cell death was induced by UV-irradiation and/or bee venom for mammalian (pancreatic cancer, metastatic breast cancer and mouse fibroblasts) and microalgae cells (Chlorella vulgaris), respectfully. Findings postulated that automated cell counter and fluorescence microscopy revealed similar patterns for the detection by both counterstain and trypan blue in mammalian cells. Interestingly, flow cytometry did provide an accurate and significant detection for only one mammalian cell line when UV-treatment was followed by routine Annexin V-Propidium iodide counterstaining. Unlike, only flow cytometry revealed a significant change in the detection of death of microalgae cells by Annexin V-Propidium iodide method, but both Annexin and conventional trypan blue methods were not applicable for the automated cell counter and microscopic detections for microalgae cells. The related outputs propose that the obtaining reliable quantitation strongly depends on cell type and instruments used. These suggest the necessity of optimization and validation endeavors before any cell death detection initiative. The analytical outcomes present insights into detailed assessment of cell death detection of eukaryotic cells and provide a direction to researchers to consider.

Comparison of Pre- and Postoperative Sleep Studies in Patients Undergoing Sphincter Pharyngoplasty.

OBJECTIVE: To determine if sphincter pharyngoplasty changes sleep study parameters on patients undergoing surgery for velopharyngeal insufficiency (VPI). DESIGN: Retrospective chart review on patients undergoing sphincter pharyngoplasty for VPI with pre- and postoperative polysomnography completed. SETTING: Institutional study at a tertiary pediatric hospital. PATIENTS: All patients who underwent sphincter pharyngoplasty over a 20-year period were reviewed; all patients with both pre- and postoperative sleep studies were collected for evaluation. INTERVENTIONS: Sphincter pharyngoplasty for patients with VPI. MAIN OUTCOME MEASURE: Sleep study parameters collected include apnea-hypopnea index (AHI), obstructive and central apneas, hypopneas, and mixed events. The preoperative values were compared to postoperative values. RESULTS: There were 98 patients collected with sleep studies for review. Of these, 32 patients had both pre- and postoperative sleep studies. The AHI increased from 1.8 preoperatively to a postoperative value of 4.8 ( P = .004). The number of obstructive events per night went from 4.6 to 17.6 postoperatively ( P = .04). The number of hypopneas increased from 4.0 to 13.6 ( P = .003). The other parameters were not statistically different, central events decreased from 4.8 to 2.1 ( P = .086), and mixed events were essentially unchanged from 0.2 to 0.5 ( P = .17) events per night. CONCLUSIONS: Patients undergoing sphincter pharyngoplasty for VPI may experience an increase in their postoperative AHI, obstructive events per night, and hypopneas. Sleep studies should be performed both pre- and postoperatively to assess the need for intervention or change in management.

Factors Predicting Hospital Length of Stay Following Pharyngeal Flap for Velopharyngeal Insufficiency.

OBJECTIVE: Routine hospital admission following pharyngeal flap (PF) to correct velopharyngeal insufficiency (VPI) is the standard at most hospitals. Nevertheless, there is increasing resistance from third-party providers to approve stays longer than a "short stay" (23-hour) observation period. The purpose of the current study was to evaluate length of stay (LOS) and document potential influencing factors following PF. DESIGN: Retrospective chart review. Demographic and perioperative data were collected, and statistical analyses were performed to determine associations with hospital length of stay (LOS). Readiness for discharge was determined by oral intake, analgesic requirement, and respiratory status. SETTING: Tertiary care children's hospital Participants: All patients undergoing PF for VPI between 1990 and 2014. OUTCOME MEASURES: (1) LOS, (2) % satisfying all discharge criteria within a 23-hour observational time frame. RESULTS: Seventy-five patients were studied, with an average age of 6.8 years. Mean LOS was 65.4 hours. Only 11 patients (14.9%) met all discharge criteria by 23 hours. Multivariate predictors of shorter LOS were increasing patient age, male gender, lack of syndromic association, administration of an intraoperative antiemetic and steroids, and shorter anesthetic duration. Time to first oral intake correlated positively with LOS. Administration of intraoperative antiemetics increased the odds of meeting all discharge criteria within 23 hours by a factor of 12. CONCLUSIONS: Identification of factors associated with LOS after PF may allow providers to predict and potentially mitigate hospital stays. Nevertheless, most patients undergoing PF are not clinically ready for discharge within a short-stay (23-hour) observation period.