Epigenetic regulation of p63 blocks squamous-to-neuroendocrine transdifferentiation in esophageal development and malignancy.

While cell fate determination and maintenance are important in establishing and preserving tissue identity and function during development, aberrant cell fate transition leads to cancer cell heterogeneity and resistance to treatment. Here, we report an unexpected role for the transcription factor p63 (Trp63/TP63) in the fate choice of squamous versus neuroendocrine lineage in esophageal development and malignancy. Deletion of p63 results in extensive neuroendocrine differentiation in the developing mouse esophagus and esophageal progenitors derived from human embryonic stem cells. In human esophageal neuroendocrine carcinoma (eNEC) cells, p63 is transcriptionally silenced by EZH2-mediated H3K27 trimethylation (H3K27me3). Upregulation of the major p63 isoform ΔNp63α, through either ectopic expression or EZH2 inhibition, promotes squamous transdifferentiation of eNEC cells. Together these findings uncover p63 as a rheostat in coordinating the transition between squamous and neuroendocrine cell fates during esophageal development and tumor progression.

What Is the Optimal Stenting Duration After Ureteroscopy and Stone Intervention? Impact of Dwell Time on Postoperative Emergency Department Visits.

PURPOSE: AUA stone management guidelines recommend stenting duration following ureteroscopy be minimized to reduce morbidity; stents with extraction strings may be used for this purpose. However, an animal study demonstrated that short dwell time results in suboptimal ureteral dilation, and a pilot clinical study showed this increases postprocedure events. Using real-world practice data we examined stent dwell time after ureteroscopy and its association with postoperative emergency department visits. MATERIALS AND METHODS: We used the Michigan Urological Surgery Improvement Collaborative registry to identify ureteroscopy and stenting procedures (2016-2019). Pre-stented cases were excluded. Stenting cohorts with and without strings were analyzed. Using multivariable logistic regression we evaluated the risk of an emergency department visit occurring on the day of, or day after, stent removal based on dwell time and string status. RESULTS: We identified 4,437 procedures; 1,690 (38%) had a string. Median dwell time was lower in patients with a string (5 vs 9 days). Ureteroscopy in younger patients, smaller stones, or renal stone location had a higher frequency of string use. The predicted probability of an emergency department visit was significantly greater in procedures with string, compared to without string, when dwell times were less than 5 days (P < .01) but were not statistically significant after. CONCLUSIONS: Patients who had ureteroscopy and stenting with a string have short dwell times. Patients are at increased risk of a postoperative emergency department visit around the time of stent removal if dwell time is ≤4 days. We recommended stenting duration of at least 5 days in nonpre-stented patients.

The development and stem cells of the esophagus.

The esophagus is derived from the anterior portion of the foregut endoderm, which also gives rise to the respiratory system. As it develops, the esophageal lining is transformed from a simple columnar epithelium into a stratified squamous cell layer, accompanied by the replacement of unspecified mesenchyme with layers of muscle cells. Studies in animal models have provided significant insights into the roles of various signaling pathways in esophageal development. More recent studies using human pluripotent stem cells (hPSCs) further demonstrate that some of these signaling pathways are conserved in human esophageal development. In addition, a combination of mouse genetics and hPSC differentiation approaches have uncovered new players that control esophageal morphogenesis. In this Review, we summarize these new findings and discuss how the esophagus is established and matures throughout different stages, including its initial specification, respiratory-esophageal separation, epithelial morphogenesis and maintenance. We also discuss esophageal muscular development and enteric nervous system innervation, which are essential for esophageal structure and function.

Beware of the sneeze.

Switch/sucrose non-fermentable-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 (SMARCB1), also known as integrase interactor 1-deficient sinonasal carcinoma, is a rare entity that was first described in 2014. Since then, there have been 39 cases published in the literature, with basaloid or plasmacytoid/rhabdoid morphology being the most common pathological subtype. We report a patient with switch/sucrose non-fermentable-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 (integrase interactor 1)-deficient sinonasal carcinoma who had permanent vision loss after valsalva-induced acute hemorrhage and resultant orbital compartment syndrome.

Analysis of Periorbital Soft Tissue in Thyroid Eye Disease.

PURPOSE: To investigate periorbital tissue enlargement in thyroid eye disease (TED) by 3-dimensional CT volumetric analysis. METHODS: Twenty-four adult subjects, 16 with TED and 8 controls, were studied. Three-dimensional volumetric calculations were performed on CT imaging of the orbit and face, focusing on the retroorbicularis oculi fat, suborbicularis oculi fat, facial muscles in periorbital region, orbital fat, extraocular muscles, and orbital volume. Analysis was performed using JMP version 12 software. Each measure was compared between the TED and control groups using Wilcoxon rank sum test. Correlations were investigated between periorbital and orbital tissue using the Spearman's correlation coefficient method. RESULTS: A statistically significant increase in volume was measured in TED patients in the superior and inferior periorbital fat tissue (p = 0.0044, p = 0.047), including the retroorbicularis oculi fat (p = 0.0011), suborbicularis oculi fat (p = 0.0093), and a decrease in facial muscle of superior periorbital region (p = 0.035). Strong positive correlation was noted between the muscles of superior and inferior periorbital region (rs = 0.65; p = 0.0006), and between the suborbicularis oculi fat and retroorbicularis oculi fat (rs = 0.50; p = 0.013). No correlation was observed between orbital and periorbital tissue, except between the orbital fat and the inferior periorbital fat tissue (p = 0.047). CONCLUSIONS: Facial fat in the periorbital region is enlarged in TED, with the superior component correlating with orbital fat tissue expansion. These findings may assist in the clinical evaluation and management of disfigurement in TED patients.Superior and inferior periorbital fat is enlarged in thyroid eye disease with the superior periorbital fat expansion correlating with orbital fat expansion.

Single Suture Müeller Muscle Conjunctival Resection (ssMMCR): A Modified Technique for Ptosis Repair.

PURPOSE: To evaluate the efficacy in degree of ptosis correction achieved by single suture Müeller muscle conjunctival resection (ssMMCR) when compared with that of traditional MMCR. METHODS: A retrospective chart analysis of patients who underwent either ssMMCR or traditional MMCR at 2 institutions. Single suture MMCR was performed after using a ptosis clamp to imbricate conjunctiva and Müeller muscle. Margin-to-reflex distance 1 was measured pre- and postoperatively, and the change in margin-to-reflex distance 1 was analyzed for both groups. Patients were monitored in follow up for postoperative complications including lagophthalmos, corneal abrasions, and change in visual acuity. Statistical analysis was performed using the Microsoft Excel and Stata software programs. RESULTS: Twenty-seven and 30 patients underwent single suture and traditional MMCR, respectively. The ssMMCR and MMCR groups were followed postoperatively for approximately 4.2 and 9.7 months, respectively and the average margin-to-reflex distance 1 increased by 2.93 mm and 2.81 mm, respectively. Notably, there was no statistically significant difference in the means identified by t test. Of the 94 eyelid surgeries evaluated, 1 ssMMCR and 3 MMCR eyelids required further surgical revision for persistent ptosis, and 1 ssMMCR developed a transient corneal epithelial abrasion. Surgical revisions were rare with both procedures, and patient satisfaction was high. CONCLUSIONS: Single suture MMCR is an efficient and effective method for ptosis repair. It results in comparable outcomes including elevation in margin-to-reflex distance 1, safety profile, and reoperation rates, when compared with traditional MMCR.

Accurate definition of control strategies using cross validated stepwise regression and Monte Carlo simulation.

Drug manufacturing processes must consistently deliver safe and effective product. A key part of achieving this is process validation utilizing Quality by Design (QbD) principles. To meet process validation requirements, process characterization (PC) studies are often performed to expand process understanding and establish an appropriate control strategy that enables the manufacturing process to consistently deliver a target product profile. Two key elements of the control strategy resulting from PC work are a list of critical process parameters (CPPs) and defined operating ranges (ORs). These are frequently derived based on mathematical models describing the relationship between process parameters and critical quality attributes (CQAs). Risk assessment and design of experiments (DOE) techniques are effectively deployed in the industry to identify parameters to study and build process understanding. However, traditional data analysis techniques do not fully utilize the data produced by these studies. In particular, stepwise regression algorithms based on p-values are prone to generate false positives and overfit data, potentially leading to unnecessarily complex control strategies. Many of the deficiencies of traditional stepwise regression can be alleviated by applying cross validation to stepwise regression algorithms, as well as Monte Carlo simulations to estimate model accuracy and predict CQA distributions. These methods can greatly enhance process understanding and assist in the selection of CPPs. A series of PC studies were performed in bioreactors to evaluate a process to produce a recombinant monoclonal antibody. The studies examined process parameters such as dissolved oxygen, pH, temperature, inoculation density, as well as cell density at two key process steps. The resulting data were analyzed using several Monte Carlo based methods. First, cross validation was used to determine model size and select parameters to be included in the model. Next, Monte Carlo cross validation was used to compare the accuracy of different models. Finally, simulated CQA profiles were generated to validate proposed ORs. This workflow provides greater process understanding based on a given PC data set and provides higher statistical confidence in both CPP selection and establishment of a control strategy.

Complete Excision of a Simple Dacryops Using Fibrin Sealant and Trypan Blue Mixture.

A 69-year-old woman presented to the oculofacial plastic service with a painless superotemporal subconjunctival mass in the OS. Over the past year, the lesion had been progressively enlarging, resulting in horizontal diplopia with lateral gaze. Visual acuity was within normal limits with no evidence of optic neuropathy. On examination, the lesion was tense, transilluminated, and was clinically consistent with a simple dacryops. Complete excision of the lesion was planned under local anesthesia with monitored care. To facilitate complete removal of the lesion, fibrinogen and a mixture of thrombin and trypan blue were injected to fill the cyst cavity. This blue-stained fibrin clot allowed for easy visualization of the border and ensured complete excision without collateral damage to surrounding normal tissue. Simple dacryops is often difficult to remove completely with its capsule intact and this technique allows for clear delineation of the cyst and preservation of epithelial integrity for complete and efficient removal.

Do Illuminated Foot Pedals Improve the Speed and Accuracy of Pedal Activation During Endoscopic Procedures?

PURPOSE: Endourologic procedures such as percutaneous nephrolithotomy (PCNL) employ the use of foot pedals in low-light operating room (OR) settings. These pedals can be especially difficult to locate or distinguish when several pedals are present during a single operation. Improper instrument activation in the OR has led to serious complications ranging from unintentional electrocautery to patient burns and even an intraoperative explosion. This study evaluates the impact of color-coded illumination on speed and efficiency of foot pedal activation. MATERIALS AND METHODS: During a simulated PCNL procedure, the foot pedals for a C-arm, laser, and ultrasonic lithotripter (USL) were placed in random positions. Ten participants performed pedal activation in a randomized sequence. Objective outcomes included time to instrument activation, number of attempted pedal presses, number of incomplete pedal presses, and number of incorrect pedal presses. Subjective preferences for pedal illumination were also determined. Data were analyzed using Mann-Whitney U, Wilcoxon signed-rank, and Chi-square tests with p < 0.05 indicating statistical significance. RESULTS: Illuminated foot pedals were associated with decreases in the average activation time for all instruments collectively (3.95 seconds vs 6.49 seconds; p = 0.017) and individually (C-arm: 3.07 seconds vs 4.21 seconds; p = 0.006; laser: 13.04 seconds vs 15.18 seconds; p < 0.001; USL: 3.28 seconds vs 4.91 seconds; p < 0.001) compared with nonilluminated pedals. Illuminated pedals were associated with fewer attempted pedal presses (33.5 vs 39.5; p = 0.007) and incomplete pedal presses (1.5 vs 8.5; p = 0.002). The number of incorrect pedal presses decreased with illumination, but this did not reach statistical significance (0 vs 0.5; p = 0.08). Participants reported that illumination simplified pedal activation and recommended its use (p < 0.01). CONCLUSION: Color-coded illumination improved the speed and efficiency of foot pedal activation during simulated PCNL. Participants subjectively preferred using illuminated foot pedals for endourologic procedures and felt that they improved safety and efficiency.

Suture constructs for rehabilitation using early active motion after tendon transfer surgery.

The purpose of this study was to better define an ideal tendon transfer suture construct to allow for early active range of motion. A side-to-side tendon construct was used to test suture technique (cross stich vs. Krackow stitch), number of suture throws, and calibre of suture. A minimum load to failure of 100 N was used to comfortably allow early motion while minimizing rupture risk. All constructs tested, except the 4-0 Krackow construct, were strong enough to withstand 100 N of load. The choice of suture should be based on surgeon preference, patient compliance, and specific surgery, and 3-0 non-absorbable suture may be more suitable for tendon transfers from a yield force standpoint.

Management of Severe Cicatricial Entropion With Labial Mucous Membrane Graft in Cicatricial Ocular Surface Disorders.

The management of cicatricial entropion represents a therapeutic challenge especially when the underlying causes are progressive cicatricial diseases that affect the ocular surface. The authors aimed to report long-term efficacy of labial mucous membrane graft to manage severe cicatricial entropion of the upper eyelid. This study is a retrospective chart review of patients who underwent tarsotomy associated with labial mucous membrane graft to treat severe cicatricial entropion of the upper eyelid. Surgeries were performed over a 16-year period. Clinical data (age, gender, etiology of the cicatricial entropion, improvement of symptoms, eyelid position, recurrence, complications, and follow-up period) were extracted from these patients' charts. Etiology of the cicatricial entropion, improvement of symptoms, eyelid position, recurrence, complications, and follow-up period were evaluated. Sixty-three eyelids from 44 patients underwent surgery. Mean follow-up was 48.4 ±â€Š46.1 months (range 6 months to 15 years). Main underlying diagnoses were Stevens-Johnson syndrome (63%), trachoma (19%), chemical injury (8%), and trauma (5%). Forty-three patients (98%) reported improvement of ocular symptoms after the procedure. Complete resolution (restoration of the upper eyelid margin to normal anatomic position with good esthetic appearance) was achieved in 52 eyelids (83%). Recurrence occurred in 7 (11%) eyelids. No postoperative infection, failure of graft survival, or other complications were observed. The use of labial mucous membrane as a posterior lamella graft showed good functional and cosmetic outcomes, long-term stability and low recurrence rates in the treatment of severe cicatricial entropion of the upper eyelid.

Avian Influenza A(H7N2) Virus in Human Exposed to Sick Cats, New York, USA, 2016.

An outbreak of influenza A(H7N2) virus in cats in a shelter in New York, NY, USA, resulted in zoonotic transmission. Virus isolated from the infected human was closely related to virus isolated from a cat; both were related to low pathogenicity avian influenza A(H7N2) viruses detected in the United States during the early 2000s.

Jackets Off: The Impact of Laser Fiber Stripping on Power Output and Stone Degradation.

PURPOSE: To investigate the effect of laser fiber stripping on stone fragmentation and laser fiber power output. MATERIALS AND METHODS: In a benchtop simulation of laser lithotripsy, 20 BegoStone phantoms were positioned within a ureteral model and irradiated for 10 minutes at 8 Hz and 0.8 J. A freshly cleaved 365 µm laser fiber was used for all trials, with half of the fibers also undergoing stripping. Power output was measured at 1-minute intervals, beginning with an initial prelithotripsy recording at 0 minutes. Fiber tips were imaged with scanning electron microscopy. In a single-blinded manner, final masses of residual stone fragments were measured and used to quantify stone breakdown. Independent-sample Mann-Whitney U tests were performed with significance set at p < 0.05, comparing stripped and unstripped fiber tips with respect to power output and fraction of stone fragmentation. RESULTS: Mean power output after 1 minute of lasing was significantly greater in unstripped laser fibers (p = 0.015), while fibers, whether stripped or not, demonstrated no significant output differences prelithotripsy or at any time from 2 to 10 minutes. However, stripped laser fibers achieved significantly increased stone breakdown compared to unstripped fibers (p = 0.004), fragmenting 63 mg (25%) more of the initial stone mass per trial. CONCLUSIONS: Although unstripped laser fibers provided superior power output at 1 minute, output at all other time points was similar between stripped and unstripped fibers. However, despite similar optical output, stripped laser fibers achieved greater stone fragmentation, possibly due to improved contact between stone and fiber tip.

Upper Eyelid Blepharoplasty: Evaluation, Treatment, and Complication Minimization.

Upper eyelid blepharoplasty is one of the most common procedures performed worldwide for both functional and cosmetic indications. There is a high rate of patient satisfaction; however, in this era of social media, patient expectations are higher than ever. Today's digitally savvy patients expect perfect outcomes with no complications and rapid recovery. To achieve optimal results, a careful preoperative evaluation and sound surgical technique is essential for minimizing complications. Here the authors review their approach to the management of the blepharoplasty patient.

Percutaneous Externally Assembled Laparoscopic vs Laparoendoscopic Single-Site Nephrectomy in a Porcine Model: A Prospective, Randomized, Blinded, Study.

OBJECTIVE: To compare the outcomes between laparoendoscopic single-site (LESS) and a novel percutaneous externally assembled laparoscopic (PEAL) nephrectomy in an in vivo porcine model. MATERIALS AND METHODS: Ten female farm pigs were randomized to LESS nephrectomy (5) or PEAL nephrectomy (5). Operative times, estimated blood loss, and intraoperative and postoperative complications were compared. The surgeons used a Likert scale to grade difficulty of peritoneal access, port placement, tool assembly, hilar dissection, closure, and overall difficulty of surgery. Scar assessment was performed by a blinded plastic surgeon using the Vancouver Scar Scale. Descriptive statistics were reported as median and range. The Mann-Whitney U test was used for continuous and ordinal variables. A p value <0.05 was considered significant. RESULTS: Median operative time was significantly shorter in the PEAL group vs the LESS group (85 minutes vs 127 minutes, p = 0.03). Median Likert scores showed overall hilar dissection and nephrectomy to be significantly easier using PEAL compared with LESS (2 vs 9, p < 0.01 for both). The PEAL instruments left no visible scar at 5 feet in any animal, and only 1 out of 10 scars could be identified on physical examination. CONCLUSIONS: The PEAL surgical paradigm demonstrates nearly scarless outcomes while providing shorter operative times and easier performance than LESS nephrectomy in a porcine model.

Establishment of prostate cancer spheres from a prostate cancer cell line after phenethyl isothiocyanate treatment and discovery of androgen-dependent reversible differentiation between sphere and neuroendocrine cells.

Prostate cancer can transform from androgen-responsive to an androgen-independent phenotype. The mechanism responsible for the transformation remains unclear. We studied the effects of an epigenetic modulator, phenethyl isothiocyanate (PEITC), on the androgen-responsive LNCaP cells. After treatment with PEITC, floating spheres were formed with characteristics of prostate cancer stem cells (PCSC). These spheres were capable of self-renewal in media with and without androgen. They have been maintained in both types of media as long term cultures. Upon androgen deprivation, the adherent spheres differentiated to neuroendocrine cells (NEC) with decreased proliferation, expression of androgen receptor, and PSA. NEC reverse differentiated to spheres when androgen was replenished. The sphere cells expressed surface marker CD44 and had enhanced histone H3K4 acetylation, DNMT1 down-regulation and GSTP1 activation. We hypothesize that PEITC-mediated alteration in epigenomics of LNCaP cells may give rise to sphere cells, whereas reversible androgenomic alterations govern the shuttling between sphere PCSC and progeny NEC. Our findings identify unrecognized properties of prostate cancer sphere cells with multi-potential plasticity. This system will facilitate development of novel therapeutic agents and allow further exploration into epigenomics and androgenomics governing the transformation to hormone refractory prostate cancer.

Prospective Comparative Analysis of 4 Different Intraocular Pressure Measurement Techniques and Their Effects on Pressure Readings.

PURPOSE: To compare intraocular pressure (IOP) measurement using the Goldmann applanation tonometry (GAT) without fluorescein, with fluorescein strips, with fluorescein droplets, and IOP measurement with Tono-Pen Avia (TPA). PATIENTS AND METHODS: This was a prospective comparative clinical analysis. It was performed in clinical practice. The study population consisted of 40 volunteer patients, 1 eye per patient. All patients who were 18 years and older having routine ophthalmological examination were eligible to participate. Active corneal abrasions and/or ulcers, previous glaucoma surgery, or prostheses interfering with GAT measurement were excluded. GAT IOP was measured first without fluorescein, then with fluorescein strip, then with fluorescein droplet, and finally with the TPA device. The main outcome measure was central corneal IOP. RESULTS: Mean±SD IOP measurements for GAT without fluorescein, with fluorescein strip, with fluorescein droplet, and for TPA groups were 12.65±3.01, 14.70±2.82, 15.78±2.64, and 16.33±3.08 mm Hg, respectively. Repeated-measures analysis of variance corrected with the Greenhouse-Geisser estimate ([Latin Small Letter Open E]=0.732) showed that measuring technique had a significant effect on IOP measurements (F2.20,85.59=34.66, P<0.001). The pairwise post hoc testing showed statistically significant mean differences (P≤0.001) between all techniques except when GAT with fluorescein droplet was compared with TPA (P=0.222). The Bland-Altman analyses showed 95% limits of agreement maximum potential discrepancies in measurement ranging from 5.89 mm Hg in the GAT with fluorescein strip versus droplet compared with 11.83 mm Hg in the GAT with fluorescein strip versus TPA comparison. CONCLUSIONS: IOP measurement technique significantly impacted the values obtained. The ophthalmologist should ensure consistent measurement technique to minimize variability when following patients.