A novel role for trithorax in the gene regulatory network for a rapidly evolving fruit fly pigmentation trait.

Animal traits develop through the expression and action of numerous regulatory and realizator genes that comprise a gene regulatory network (GRN). For each GRN, its underlying patterns of gene expression are controlled by cis-regulatory elements (CREs) that bind activating and repressing transcription factors. These interactions drive cell-type and developmental stage-specific transcriptional activation or repression. Most GRNs remain incompletely mapped, and a major barrier to this daunting task is CRE identification. Here, we used an in silico method to identify predicted CREs (pCREs) that comprise the GRN which governs sex-specific pigmentation of Drosophila melanogaster. Through in vivo assays, we demonstrate that many pCREs activate expression in the correct cell-type and developmental stage. We employed genome editing to demonstrate that two CREs control the pupal abdomen expression of trithorax, whose function is required for the dimorphic phenotype. Surprisingly, trithorax had no detectable effect on this GRN's key trans-regulators, but shapes the sex-specific expression of two realizator genes. Comparison of sequences orthologous to these CREs supports an evolutionary scenario where these trithorax CREs predated the origin of the dimorphic trait. Collectively, this study demonstrates how in silico approaches can shed novel insights on the GRN basis for a trait's development and evolution.

Appendicitis and Cholecystitis in Pregnancy.

Up to 2% of pregnant women develop a disease that requires nonobstetrical operative intervention during pregnancy. We discuss the issues unique to pregnant patients as they pertain to the presentation, diagnosis, and management of nonobstetric surgical disease, with an emphasis on 2 of the most common diseases that affect pregnant women: appendicitis and cholecystitis. Surgery has been demonstrated to be safe and effective during pregnancy, provided proper precautions are taken into account. It is the consensus of multiple professional committees and societies that no pregnant women should be delayed or denied a necessary surgery because of pregnancy.

SMAD2 and p38 signaling pathways act in concert to determine XY primordial germ cell fate in mice.

The sex of primordial germ cells (PGCs) is determined in developing gonads on the basis of cues from somatic cells. In XY gonads, sex-determining region Y (SRY) triggers fibroblast growth factor 9 (FGF9) expression in somatic cells. FGF signaling, together with downstream nodal/activin signaling, promotes male differentiation in XY germ cells by suppressing retinoic acid (RA)-dependent meiotic entry and inducing male-specific genes. However, the mechanism by which nodal/activin signaling regulates XY PGC fate is unknown. We uncovered the roles of SMAD2/3 and p38 MAPK, the putative downstream factors of nodal/activin signaling, in PGC sexual fate decision. We found that conditional deletion of Smad2, but not Smad3, from XY PGCs led to a loss of male-specific gene expression. Moreover, suppression of RA signaling did not rescue male-specific gene expression in Smad2-mutant testes, indicating that SMAD2 signaling promotes male differentiation in a RA-independent manner. By contrast, we found that p38 signaling has an important role in the suppression of RA signaling. The Smad2 deletion did not disrupt the p38 signaling pathway even though Nodal expression was significantly reduced, suggesting that p38 was not regulated by nodal signaling in XY PGCs. Additionally, the inhibition of p38 signaling in the Smad2-mutant testes severely impeded XY PGC differentiation and induced meiosis. In conclusion, we propose a model in which p38 and SMAD2 signaling coordinate to determine the sexual fate of XY PGCs.

Predicting the Risk of Postoperative Respiratory Failure in Elective Abdominal and Vascular Operations Using the National Surgical Quality Improvement Program (NSQIP) Participant Use Data File.

OBJECTIVE: This study aims to develop a Respiratory Failure Risk Score (RFRS) with good predictability for elective abdominal and vascular patients to be used in the outpatient setting for risk stratification and to guide preoperative pulmonary optimization. SUMMARY BACKGROUND DATA: Postoperative respiratory failure (RF), defined as ventilator dependency for more than 48 hours or unplanned reintubation within 30 days, is associated with increased mortality and hospital costs. Many tools have been previously described for risk stratification, but few target elective surgical candidates. METHODS: Our training sample included patients undergoing inpatient, nonemergent general and vascular procedures sampled for the American College of Surgeon National Surgical Quality Improvement Program 2012 Participant Use File. Multivariable logistic regression identified independent preoperative risk factors associated with RF, used to derive a weighted RFRS. We then determined goodness-of-fit and optimal cutoff values through receiver operator characteristic analysis and Youden indices to evaluate internal and external validity with a retrospective institutional validation sample (2013 and 2014). RESULTS: Multivariable analysis of 151,700 patients from the National Surgical Quality Improvement Program Participant Use File identified 12 variables independently associated with RF. The RFRS showed good external prediction in the validation sample with a c-statistic of 0.73 (95% confidence interval, 0.68-0.79). With the highest Youden index, 30 was determined to be the optimal cutoff value with a sensitivity 0.62 and specificity of 0.75. Additional cutoff values of 15 and 40 optimized sensitivity (>0.80) and specificity (>0.80), respectively. CONCLUSIONS: In the preoperative setting, the RFRS can effectively stratify patients into low (<15), moderate low (15-29), moderate high (30-39), and high risk (>39) to assist in patient counseling and guide application of perioperative pulmonary optimization measures.

Topologically protected states in one-dimensional continuous systems and Dirac points.

We study a class of periodic Schrödinger operators on ℝ that have Dirac points. The introduction of an "edge" via adiabatic modulation of a periodic potential by a domain wall results in the bifurcation of spatially localized "edge states," associated with the topologically protected zero-energy mode of an asymptotic one-dimensional Dirac operator. The bound states we construct can be realized as highly robust transverse-magnetic electromagnetic modes for a class of photonic waveguides with a phase defect. Our model captures many aspects of the phenomenon of topologically protected edge states for 2D bulk structures such as the honeycomb structure of graphene.

Successful Reinnervation of the Diaphragm After Intercostal to Phrenic Nerve Neurotization in Patients With High Spinal Cord Injury.

OBJECTIVE: Our objective in this study was to extend diaphragmatic pacing therapy to include paraplegic patients with high cervical spinal cord injuries between C3 and C5. INTRODUCTION: Diaphragmatic pacing has been used in patients experiencing ventilator-dependent respiratory failure due to spinal cord injury as a means to reduce or eliminate the need for mechanical ventilation. However, this technique relies on intact phrenic nerve function. Recently, phrenic nerve reconstruction with intercostal nerve grafting has expanded the indications for diaphragmatic pacing. Our study aimed to evaluate early outcomes and efficacy of intercostal nerve transfer in diaphragmatic pacing. METHODS: Four ventilator-dependent patients with high cervical spinal cord injuries were selected for this study. Each patient demonstrated absence of phrenic nerve function via external neck stimulation and laparoscopic diaphragm mapping. Each patient underwent intercostal to phrenic nerve grafting with implantation of a phrenic nerve pacer. The patients were followed, and ventilator dependence was reassessed at 1 year postoperatively. RESULTS: Our primary outcome was measured by the amount of time our patients tolerated off the ventilator per day. We found that all 4 patients have tolerated paced breathing independent of mechanical ventilation, with 1 patient achieving 24 hours of tracheostomy collar. CONCLUSIONS: From this study, intercostal to phrenic nerve transfer seems to be a promising approach in reducing or eliminating ventilator support in patients with C3 to C5 high spinal cord injury.

Somatic mosaicism for a lethal TRPV4 mutation results in non-lethal metatropic dysplasia.

Dominant mutations in TRPV4, which encodes the Transient Receptor Potential Cation Channel Subfamily V Member 4 calcium channel, result in a series of musculoskeletal disorders that include a set of peripheral neuropathies and a broad phenotypic spectrum of skeletal dysplasias. The skeletal phenotypes range from brachyolmia, in which there is scoliosis with mild short stature, through perinatal lethal metatropic dysplasia. We describe a case with phenotypic findings consistent with metatropic dysplasia, but in whom no TRPV4 mutation was detected by Sanger sequence analysis. Exome sequence analysis identified a known lethal metatropic dysplasia mutation, TRPV4L618P , which was present at lower frequency than would be expected for a heterozygous change. The affected individual was shown to be a somatic mosaic for the mutation, providing an explanation for the milder than expected phenotype. The data illustrate that high-throughput sequencing of genomic DNA can facilitate detection of mosaicism with higher sensitivity than Sanger sequence analysis and identify a new genetic mechanism for metatropic dysplasia. © 2016 Wiley Periodicals, Inc.

The role of nature-based infrastructure (NBI) in coastal resiliency planning: A literature review.

The use of nature-based infrastructure (NBI) has attracted increasing attention in the context of protection against coastal flooding. This review is focused on NBI approaches to improve coastal resilience in the face of extreme storm events, including hurricanes. We not only consider the role of NBI as a measure to protect people and property but also in the context of other ecological goods and services provided by tidal wetlands including production of fish and shellfish. Although the results of many studies suggest that populated areas protected by coastal marshes were less likely to experience damage when exposed to the full force of storm surge, it was absolutely critical to place the role of coastal wetlands into perspective by noting that while tidal marshes can reduce wave energy from low-to-moderate-energy storms, their capacity to substantially reduce storm surge remains poorly quantified. Moreover, although tidal marshes can reduce storm surge from fast moving storms, very large expanses of habitat are needed to be most effective, and for most urban settings, there is insufficient space to rely on nature-based risk reduction strategies alone. The success of a given NBI method is also context dependent on local conditions, with potentially confounding influences from substrate characteristics, topography, near shore bathymetry, distance from the shore and other physical factors and human drivers such as development patterns. Furthermore, it is important to better understand the strengths and weaknesses of newly developed NBI projects through rigorous evaluations and characterize the local specificities of the particular built and natural environments surrounding these coastal areas. In order for the relevant science to better inform policy, and assist in land-use challenges, scientists must clearly state the likelihood of success in a particular circumstance and set of conditions. We conclude that "caution is advised" before selecting a particular NBI method as there is no "one size fits all" solution to address site-specific conditions.

Follicle-stimulating hormone synthesis and fertility are intact in mice lacking SMAD3 DNA binding activity and SMAD2 in gonadotrope cells.

The activin/inhibin system regulates follicle-stimulating hormone (FSH) synthesis and release by pituitary gonadotrope cells in mammals. In vitro cell line data suggest that activins stimulate FSH ß-subunit (Fshb) transcription via complexes containing the receptor-regulated SMAD proteins SMAD2 and SMAD3. Here, we used a Cre-loxP approach to determine the necessity for SMAD2 and/or SMAD3 in FSH synthesis in vivo. Surprisingly, mice with conditional mutations in both Smad2 and Smad3 specifically in gonadotrope cells are fertile and produce FSH at quantitatively normal levels. Notably, however, we discovered that the recombined Smad3 allele produces a transcript that encodes the entirety of the SMAD3 C-terminal Mad homology 2 (MH2) domain. This protein behaves similarly to full-length SMAD3 in Fshb transcriptional assays. As the truncated protein lacks the N-terminal Mad homology 1 (MH1) domain, these results show that SMAD3 DNA-binding activity as well as SMAD2 are dispensable for normal FSH synthesis in vivo. Furthermore, the observation that deletion of proximal exons does not remove all SMAD3 function may facilitate interpretation of divergent phenotypes previously described in different Smad3 knockout mouse lines.

Pten in stromal fibroblasts suppresses mammary epithelial tumours.

The tumour stroma is believed to contribute to some of the most malignant characteristics of epithelial tumours. However, signalling between stromal and tumour cells is complex and remains poorly understood. Here we show that the genetic inactivation of Pten in stromal fibroblasts of mouse mammary glands accelerated the initiation, progression and malignant transformation of mammary epithelial tumours. This was associated with the massive remodelling of the extracellular matrix (ECM), innate immune cell infiltration and increased angiogenesis. Loss of Pten in stromal fibroblasts led to increased expression, phosphorylation (T72) and recruitment of Ets2 to target promoters known to be involved in these processes. Remarkably, Ets2 inactivation in Pten stroma-deleted tumours ameliorated disruption of the tumour microenvironment and was sufficient to decrease tumour growth and progression. Global gene expression profiling of mammary stromal cells identified a Pten-specific signature that was highly represented in the tumour stroma of patients with breast cancer. These findings identify the Pten-Ets2 axis as a critical stroma-specific signalling pathway that suppresses mammary epithelial tumours.

Addressing Consent Issues in Donation After Circulatory Determination of Death.

Given the widening gap between the number of individuals on transplant waiting lists and the availability of donated organs, as well as the recent plateau in donations based on neurological criteria (i.e., brain death), there has been a growing interest in expanding donation after circulatory determination of death. While the prevalence of this form of organ donation continues to increase, many thorny ethical issues remain, often creating moral distress in both clinicians and families. In this article, we address one of these issues, namely, the challenges surrounding patient and surrogate informed consent for donation after circulatory determination of death. First we discuss several general concerns regarding consent related to this form of organ donation, and then we address additional issues that are unique to three different patient categories: adult patients with medical decision-making capacity or potential capacity, adult patients who lack capacity, and pediatric patients.

The effect of vertebral fracture on the early neurologic recovery in patients with central cord syndrome.

PURPOSE: To compare early changes in the ASIA Motor Score (AMS) between patients with central cord syndrome (CCS) from an acute fracture to patients without a fracture. METHODS: Patients with CCS were identified and stratified based on the presence of a fracture. The AMS through the first week of the patients' hospitalization was obtained. Initial injury severity as well as early neurologic recovery was measured using the AMS. Analysis of variance was performed to determine if age, gender, rectal tone at presentation, congenital stenosis, or surgery within 24 h significantly effected the change in AMS. RESULTS: A strong trend (p = 0.0504) towards a more severe initial neurologic injury in patients with a fracture (AMS 59.7) than in patients without a fracture (AMS 70.2) was identified. However, in the week after injury, patients with a fracture had an improvement in their neurologic function (ΔAMS +4.8) while patients without a fracture demonstrated neurologic decline (ΔAMS -5.9). The change in AMS between patients with and without a fracture was nearly significant (p = 0.06). CONCLUSION: Patients with central cord syndrome present with similar symptoms, but injuries with and without a fracture may be associated with a different early neurologic recovery. Patients with a fracture have a more severe injury at initial presentation, but tend to have neurologic improvement in the first week; conversely patients without a fracture have a less severe initial neurologic injury, but tend to have a slight decline in neurologic function over the first week.

A Second Opinion: A Case Narrative on Clinical Ethics Mediation.

Contrasting traditional and common forms if ethics consultation with bioethics mediation. I describe the case of a "second opinion" consultation in the care of a patient with advanced cancer for whom treatment was futile. While the initial ethics consultation, performed by a colleague, let to a recommendation that some may deem ethical, the process failed to involve key stakeholders and failed to explore the underlying values and reasons for the opinions voiced by various stakeholders. The process of mediation ultimately led to creative solutions in which all stakeholders could reach consensus on a plan of care.

A Second Opinion: A Case Narrative on Clinical Ethics Mediation.

Contrasting traditional and common forms of ethics consultation with bioethics mediation, I describe the case of a "second opinion" consultation in the care of a patient with advanced cancer for whom treatment was futile. While the initial ethics consultation, performed by a colleague, led to a recommendation that some may deem ethical, the process failed to involve key stakeholders and failed to explore the underlying values and reasons for the opinions voiced by various stakeholders. The process of mediation ultimately led to creative solutions in which all stakeholders could reach consensus on a plan of care.