Neuregulin-3 Regulates Epithelial Progenitor Cell Positioning and Specifies Mammary Phenotype

Mutation of Neuregulin-3 (Nrg3) results in defective embryonic mammary gland development. Here, we investigate functions of Nrg3 signaling in embryonic mammary morphogenesis. Nrg3 regulates the distribution of epithelial progenitor cells within the presumptive mammary-forming region during early mammary morphogenesis. Basal and suprabasal epithelial cells are significantly smaller within the hypoplastic mammary primordium (MP) that forms in Nrg3 mutants, indicative of failure to acquire mammary epithelial cell (MEC) morphological phenotype. Activation of Erbb4 JM-a CYT-1, an Erbb4 isoform expressed in the developing MP, leads to MEC spreading and migration. Nrg3 promotes the accumulation of epithelial progenitor cells at the MP site in embryo explant cultures. Our results implicate Nrg3 signaling in mediating key events of mammary mesenchyme specification, including mesenchymal condensation, mitosis, and induction of mammary marker expression. Taken together, our results show Nrg3 has a major role in conferring specification of the mammary phenotype to both epithelial and mesenchymal progenitor cells

Intraoperative Evaluation of Nasal Valve Repair Interventions: A Prospective Analysis

Objectives: To allow for early identification and treatment of inadequate nasal valve repair interventions in the intraoperative setting, based on degree of nasal valve collapse quantified by suction-assisted pressure readings. Patient outcomes were measured by comparison of pre- and post-operative Nasal Obstruction Symptom Evaluation (NOSE) surveys. Study Design: Prospective study. Methods: All enrolled patients undergo suction-assisted evaluation of nasal valve collapse before surgical intervention. Patients randomized into the experimental group underwent repeat assessment after various nasal valve interventions, compared to a control group where adequacy of interventions was assessed by palpation of the nasal ala. Results: 20 patients who underwent nasal valve repair were first randomized into control (10) or experimental (10) groups. Two patients in the control group did not receive nasal valve work due to pre-operative readings and were excluded from further analysis. Nasal valve interventions included alar rim grafts (5), spreader grafts (10), batten grafts (2), and nasal valve suture suspension (8). After nasal valve interventions, average suction reading at first sign of collapse increased by 92% (p \u3c 0.0001) and average suction reading at maximal collapse increased by 16% (p \u3c 0.0001). Pre-operative NOSE scores decreased by 55% (p \u3c 0.0001) at the first follow-up visit at 9.3±3.5 days. No patients in the experimental group required additional nasal valve interventions after repeat suction-assisted evaluations intraoperatively. Conclusion: Intraoperative suction-assisted evaluation of nasal valve collapse can help assess adequacy of nasal valve interventions and determine whether additional interventions are necessary to improve nasal valve integrity.https://jdc.jefferson.edu/otoposters/1011/thumbnail.jp

Fat Graft for Parotidectomy Defect Reconstruction in the Setting of Malignant Disease

Objectives: Currently, limited data examines the safety of utilizing fat transfers in the setting of malignant parotid disease. Here we evaluate the safety of fat graft reconstruction of parotidectomy defects in the setting of malignant disease. Study Design: Retrospective cohort study Methods: Electronic chart review of patients who underwent parotidectomy from 2012-2020 were reviewed. Results: Three hundred and sixty-one patients were identified at a single institution who underwent parotidectomy, and 113 (31.3%) were for malignancy. One hundred and thirty-two patients underwent fat graft reconstruction (49.2%, n=65 for umbilical, 50.8%, n=67 for dermal). One-third of patients had malignant pathology (34.8%, n=46). The most common malignant tumors were squamous cell carcinoma (n=15), acinic cell carcinoma (n=9), and mucoepidermoid carcinoma (n=6). Twenty patients (45.5%) received postoperative radiation therapy. Complications included: surgical site necrosis (13%), hematoma (4.3%), and infection (2.2%). Overall incidence of malignant recurrence was 4.4% with a mean time of follow-up of 10.3 (range 0 – 77.3) months. Incidence of malignant recurrence in the fat graft reconstruction subset was 0% with a mean follow-up of 9.8 (range 0.2 – 49.3) months. There was no association with use of fat graft and recurrence (p\u3e0.05). Conclusion: Parotidectomy defects for malignant neoplasms can be reconstructed with fat graft transfers with no impact on surveillance for disease recurrence.https://jdc.jefferson.edu/otoposters/1010/thumbnail.jp

Sox9 regulates cell state and activity of embryonic mouse mammary progenitor cells.

Embryonic mammary cells are a unique population comprised of undifferentiated, highly plastic progenitor cells that create normal mammary tissues. The mammary gland continues to develop after birth from descendants of embryonic mammary cells. Here, we establish cell lines from mouse mammary organs, immediately after they formed during prenatal development, to facilitate studies of primitive mammary cells, which are difficult to isolate in sufficient quantities for use in functional experiments. We show that some lines can be induced to secrete milk, a distinguishing feature of mammary epithelial cells. Targeted deletion of Sox9, from one clone, decreases the ability to respond to lactogenic stimuli, consistent with a previously identified role for Sox9 in regulating luminal progenitor function. Sox9 ablation also leads to alterations in 3D morphology and downregulation of Zeb1, a key epithelial-mesenchymal transition regulator. Prenatal mammary cell lines are an invaluable resource to study regulation of mammary progenitor cell biology and development

A Sox2–Sox9 signalling axis maintains human breast luminal progenitor and breast cancer stem cells

Increased cancer stem cell content during development of resistance to tamoxifen in breast cancer is driven by multiple signals, including Sox2-dependent activation of Wnt signalling. Here, we show that Sox2 increases and estrogen reduces the expression of the transcription factor Sox9. Gain and loss of function assays indicate that Sox9 is implicated in the maintenance of human breast luminal progenitor cells. CRISPR/Cas knockout of Sox9 reduces growth of tamoxifen-resistant breast tumours in vivo. Mechanistically, Sox9 acts downstream of Sox2 to control luminal progenitor cell content and is required for expression of the cancer stem cell marker ALDH1A3 and Wnt signalling activity. Sox9 is elevated in breast cancer patients after endocrine therapy failure. This new regulatory axis highlights the relevance of SOX family transcription factors as potential therapeutic targets in breast cancer

FORENSIC PAIN MEDICINE SECTION Original Research Article Societal Costs of Prescription Opioid Abuse, Dependence, and Misuse in the United Statesp me_1075 657..667

Abstract Objectives. The objective of this study was to estimate the societal costs of prescription opioid abuse, dependence, and misuse in the United States. Methods. Costs were grouped into three categories: health care, workplace, and criminal justice. Costs were estimated by 1) quantity method, which multiplies the number of opioid abuse patients by cost per opioid abuse patient; and 2) apportionment method, which begins with overall costs of drug abuse per component and apportions the share associated with prescription opioid abuse based on relative prevalence of prescription opioid to overall drug abuse. Excess health care costs per patient were based on claims data analysis of privately insured and Medicaid beneficiaries. Other data/ information were derived from publicly available survey and other secondary sources. Conclusions. The costs of prescription opioid abuse represent a substantial and growing economic burden for the society. The increasing prevalence of abuse suggests an even greater societal burden in the future

SWATH mass spectrometry as a tool for quantitative profiling of the matrisome.

Proteomic analysis of extracellular matrix (ECM) and ECM-associated proteins, collectively known as the matrisome, is a challenging task due to the inherent complexity and insolubility of these proteins. Here we present sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH MS) as a tool for the quantitative analysis of matrisomal proteins in both non-enriched and ECM enriched tissue without the need for prior fractionation. Utilising a spectral library containing 201 matrisomal proteins, we compared the performance and reproducibility of SWATH MS over conventional data-dependent analysis mass spectrometry (DDA MS) in unfractionated murine lung and liver. SWATH MS conferred a 15-20% increase in reproducible peptide identification across replicate experiments in both tissue types and identified 54% more matrisomal proteins in the liver versus DDA MS. We further use SWATH MS to evaluate the quantitative changes in matrisome content that accompanies ECM enrichment. Our data shows that ECM enrichment led to a systematic increase in core matrisomal proteins but resulted in significant losses in matrisome-associated proteins including the cathepsins and proteins of the S100 family. Our proof-of-principle study demonstrates the utility of SWATH MS as a versatile tool for in-depth characterisation of the matrisome in unfractionated and non-enriched tissues. SIGNIFICANCE: The matrisome is a complex network of extracellular matrix (ECM) and ECM-associated proteins that provides scaffolding function to tissues and plays important roles in the regulation of fundamental cellular processes. However, due to its inherent complexity and insolubility, proteomic studies of the matrisome typically require the application of enrichment workflows prior to MS analysis. Such enrichment strategies often lead to losses in soluble matrisome-associated components. In this study, we present sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH MS) as a tool for the quantitative analysis of matrisomal proteins. We show that SWATH MS provides a more reproducible coverage of the matrisome compared to data-dependent analysis (DDA) MS. We also demonstrate that SWATH MS is capable of accurate quantification of matrisomal proteins without prior ECM enrichment and fractionation, which may simplify sample handling workflows and avoid losses in matrisome-associated proteins commonly linked to ECM enrichment

Edible crabs “Go West”: migrations and incubation cycle of Cancer pagurus revealed by electronic tags

Crustaceans are key components of marine ecosystems which, like other exploited marine taxa, show seasonable patterns of distribution and activity, with consequences for their availability to capture by targeted fisheries. Despite concerns over the sustainability of crab fisheries worldwide, difficulties in observing crabs’ behaviour over their annual cycles, and the timings and durations of reproduction, remain poorly understood. From the release of 128 mature female edible crabs tagged with electronic data storage tags (DSTs), we demonstrate predominantly westward migration in the English Channel. Eastern Channel crabs migrated further than western Channel crabs, while crabs released outside the Channel showed little or no migration. Individual migrations were punctuated by a 7-month hiatus, when crabs remained stationary, coincident with the main period of crab spawning and egg incubation. Incubation commenced earlier in the west, from late October onwards, and brooding locations, determined using tidal geolocation, occurred throughout the species range. With an overall return rate of 34%, our results demonstrate that previous reluctance to tag crabs with relatively high-cost DSTs for fear of loss following moulting is unfounded, and that DSTs can generate precise information with regards life-history metrics that would be unachievable using other conventional means