A rare case of cutaneous myxoma with trichofolliculoma-like features.

Carney complex is a rare genetic disorder associated with a number of cutaneous lesions, especially cutaneous myxomas. We present a rare case of cutaneous myxoma (superficial angiomyxoma) with trichofolliculoma-like features in a patient with Carney complex, and explore how the associated histopathology provides critical context for elucidating the etiology of this benign neoplasm.

Therapeutic Targeting of KDM1A/LSD1 in Ewing Sarcoma with SP-2509 Engages the Endoplasmic Reticulum Stress Response.

Multi-agent chemotherapeutic regimes remain the cornerstone treatment for Ewing sarcoma, the second most common bone malignancy diagnosed in pediatric and young adolescent populations. We have reached a therapeutic ceiling with conventional cytotoxic agents, highlighting the need to adopt novel approaches that specifically target the drivers of Ewing sarcoma oncogenesis. As KDM1A/lysine-specific demethylase 1 (LSD1) is highly expressed in Ewing sarcoma cell lines and tumors, with elevated expression levels associated with worse overall survival (P = 0.033), this study has examined biomarkers of sensitivity and mechanisms of cytotoxicity to targeted KDM1A inhibition using SP-2509 (reversible KDM1A inhibitor). We report, that innate resistance to SP-2509 was not observed in our Ewing sarcoma cell line cohort (n = 17; IC50 range, 81 -1,593 nmol/L), in contrast resistance to the next-generation KDM1A irreversible inhibitor GSK-LSD1 was observed across multiple cell lines (IC50 > 300 µmol/L). Although TP53/STAG2/CDKN2A status and basal KDM1A mRNA and protein levels did not correlate with SP-2509 response, induction of KDM1B following SP-2509 treatment was strongly associated with SP-2509 hypersensitivity. We show that the transcriptional profile driven by SP-2509 strongly mirrors KDM1A genetic depletion. Mechanistically, RNA-seq analysis revealed that SP-2509 imparts robust apoptosis through engagement of the endoplasmic reticulum stress pathway. In addition, ETS1/HIST1H2BM were specifically induced/repressed, respectively following SP-2509 treatment only in our hypersensitive cell lines. Together, our findings provide key insights into the mechanisms of SP-2509 cytotoxicity as well as biomarkers that can be used to predict KDM1A inhibitor sensitivity in Ewing sarcoma. Mol Cancer Ther; 17(9); 1902-16. ©2018 AACR.

Conservation of Neotropical migratory birds in tropical hardwood and oil palm plantations.

Tropical forests in the Americas are undergoing rapid conversion to commercial agriculture, and many migratory bird species that use these forests have experienced corresponding populations declines. Conservation research for migratory birds in the tropics has focused overwhelmingly on shade coffee plantations and adjacent forest, but both cover types are now in decline, creating an urgent need to evaluate conservation opportunities in other agricultural systems. Here we compare how a community of 42 Neotropical migratory bird species and a subset of five conservation-priority species differ in usage and habitat associations among a secondary forest baseline and four expanding commercial plantation systems in Guatemala: African oil palm, teak, rubber, and mixed-native hardwoods. We found that mixed-native hardwood plantations supported the highest richness and diversity of all migrants and that the three hardwood plantation types generally outperformed oil palm in richness and diversity metrics. Despite this, oil palm supported high abundance of several common and widespread species also experiencing range-wide population declines and may therefore play an important role in conserving common species. Mature secondary forest hosted low abundance and diversity of the full migratory community, but high abundance and richness of conservation priority migrants along with native hardwood and teak plantations. Likewise, the percentage of forest cover on the landscape was positively associated with priority migrant abundance and richness but negatively associated with the abundance of migrants in general, highlighting how individual species within the broad group of Neotropical migratory landbirds respond differently to anthropogenic changes in land use. Across all cover types, the retention of tall overstory trees increased the abundance, richness, and diversity of all migrants, which indicates that vertical structural diversity and remnant trees are important habitat features for birds in agricultural landscapes. Our findings show that conservation opportunities exist in hardwood and oil palm plantations, though the species likely to benefit from conservation action will vary among plantation types. For the subset of conservation priority migrants, our results suggest that conservation efforts should combine strategies that retain and restore secondary forest, promote the adoption of native hardwood and teak plantations, and promote the retention of tall, remnant trees in agricultural landscapes.

Identification of two types of GGAA-microsatellites and their roles in EWS/FLI binding and gene regulation in Ewing sarcoma.

Ewing sarcoma is a bone malignancy of children and young adults, frequently harboring the EWS/FLI chromosomal translocation. The resulting fusion protein is an aberrant transcription factor that uses highly repetitive GGAA-containing elements (microsatellites) to activate and repress thousands of target genes mediating oncogenesis. However, the mechanisms of EWS/FLI interaction with microsatellites and regulation of target gene expression is not clearly understood. Here, we profile genome-wide protein binding and gene expression. Using a combination of unbiased genome-wide computational and experimental analysis, we define GGAA-microsatellites in a Ewing sarcoma context. We identify two distinct classes of GGAA-microsatellites and demonstrate that EWS/FLI responsiveness is dependent on microsatellite length. At close range "promoter-like" microsatellites, EWS/FLI binding and subsequent target gene activation is highly dependent on number of GGAA-motifs. "Enhancer-like" microsatellites demonstrate length-dependent EWS/FLI binding, but minimal correlation for activated and none for repressed targets. Our data suggest EWS/FLI binds to "promoter-like" and "enhancer-like" microsatellites to mediate activation and repression of target genes through different regulatory mechanisms. Such characterization contributes valuable insight to EWS/FLI transcription factor biology and clarifies the role of GGAA-microsatellites on a global genomic scale. This may provide unique perspective on the role of non-coding DNA in cancer susceptibility and therapeutic development.

Role for the EWS domain of EWS/FLI in binding GGAA-microsatellites required for Ewing sarcoma anchorage independent growth.

Ewing sarcoma usually expresses the EWS/FLI fusion transcription factor oncoprotein. EWS/FLI regulates myriad genes required for Ewing sarcoma development. EWS/FLI binds GGAA-microsatellite sequences in vivo and in vitro. These sequences provide EWS/FLI-mediated activation to reporter constructs, suggesting that they function as EWS/FLI-response elements. We now demonstrate the critical role of an EWS/FLI-bound GGAA-microsatellite in regulation of the NR0B1 gene as well as for Ewing sarcoma proliferation and anchorage-independent growth. Clinically, genomic GGAA-microsatellites are highly variable and polymorphic. Current data suggest that there is an optimal "sweet-spot" GGAA-microsatellite length (of 18-26 GGAA repeats) that confers maximal EWS/FLI-responsiveness to target genes, but the mechanistic basis for this remains unknown. Our biochemical studies, using recombinant Δ22 (a version of EWS/FLI containing only the FLI portion), demonstrate a stoichiometry of one Δ22-monomer binding to every two consecutive GGAA-repeats on shorter microsatellite sequences. Surprisingly, the affinity for Δ22 binding to GGAA-microsatellites significantly decreased, and ultimately became unmeasureable, when the size of the microsatellite was increased to the sweet-spot length. In contrast, a fully functional EWS/FLI mutant (Mut9, which retains approximately half of the EWS portion of the fusion) showed low affinity for smaller GGAA-microsatellites but instead significantly increased its affinity at sweet-spot microsatellite lengths. Single-gene ChIP and genome-wide ChIP-sequencing (ChIP-seq) and RNA-seq studies extended these findings to the in vivo setting. Together, these data demonstrate the critical requirement of GGAA-microsatellites as EWS/FLI activating response elements in vivo and reveal an unexpected role for the EWS portion of the EWS/FLI fusion in binding to sweet-spot GGAA-microsatellites.

Implications of inaccurate clinical nodal staging in pancreatic adenocarcinoma.

BACKGROUND: Many patients with stage I-II pancreatic adenocarcinoma do not undergo resection. We hypothesized that (1) clinical staging underestimates nodal involvement, causing stage IIB to have a greater percent of resected patients and (2) this stage-shift causes discrepancies in observed survival. METHODS: The Surveillance, Epidemiology, and End Results (SEER) research database was used to evaluate cause-specific survival in patients with pancreatic adenocarcinoma from 2004-2012. Survival was compared using the log-rank test. Single-center data on 105 patients who underwent resection of pancreatic adenocarcinoma without neoadjuvant treatment were used to compare clinical and pathologic nodal staging. RESULTS: In SEER data, medium-term survival in stage IIB was superior to IB and IIA, with median cause-specific survival of 14, 9, and 11 months, respectively (P < .001). Seventy-two percent of stage IIB patients underwent resection vs 28% in IB and 36% in IIA (P < .001). In our institutional data, 12.4% of patients had clinical evidence of nodal involvement vs 69.5% by pathologic staging (P < .001). Among clinical stage IA-IIA patients, 71.6% had nodal involvement by pathologic staging. CONCLUSION: Both SEER and institutional data support substantial underestimation of nodal involvement by clinical staging. This finding has implications in decisions regarding neoadjuvant therapy and analysis of outcomes in the absence of pathologic staging.

Induction and inhibition of CPAF activity during analysis of Chlamydia-infected cells.

Studies of the chlamydial protease CPAF have been complicated by difficulties in distinguishing bona fide intracellular proteolysis from in vitro proteolysis. This confounding issue has been attributed to CPAF activity in lysates from Chlamydia-infected cells. We compared three methods that have been used to inhibit in vitro CPAF-mediated proteolysis: (1) pre-treatment of infected cells with the inhibitor clasto-lactacystin, (2) direct cell lysis in 8 M urea and (3) direct lysis in hot 1% SDS buffer. We identified a number of experimental conditions that reduce the effectiveness of each method in preventing CPAF activity during lysate preparation. The amount of in vitro proteolysis in a lysate was variable and depended on factors such as the specific substrate and the time in the intracellular infection. Additionally, we demonstrated for the first time that artifactual CPAF activity is induced before cell lysis by standard cell detachment methods, including trypsinization. Protein analysis of Chlamydia-infected cells therefore requires precautions to inhibit CPAF activity during both cell detachment and lysate preparation, followed by verification that the cell lysates do not contain residual CPAF activity. These concerns about artifactual proteolysis extend beyond studies of CPAF function because they have the potential to affect the analyses of host and chlamydial proteins from Chlamydia-infected cells.

Clinical and biochemical function of polymorphic NR0B1 GGAA-microsatellites in Ewing sarcoma: a report from the Children's Oncology Group.

BACKGROUND: The genetics involved in Ewing sarcoma susceptibility and prognosis are poorly understood. EWS/FLI and related EWS/ETS chimeras upregulate numerous gene targets via promoter-based GGAA-microsatellite response elements. These microsatellites are highly polymorphic in humans, and preliminary evidence suggests EWS/FLI-mediated gene expression is highly dependent on the number of GGAA motifs within the microsatellite. OBJECTIVES: Here we sought to examine the polymorphic spectrum of a GGAA-microsatellite within the NR0B1 promoter (a critical EWS/FLI target) in primary Ewing sarcoma tumors, and characterize how this polymorphism influences gene expression and clinical outcomes. RESULTS: A complex, bimodal pattern of EWS/FLI-mediated gene expression was observed across a wide range of GGAA motifs, with maximal expression observed in constructs containing 20-26 GGAA motifs. Relative to white European and African controls, the NR0B1 GGAA-microsatellite in tumor cells demonstrated a strong bias for haplotypes containing 21-25 GGAA motifs suggesting a relationship between microsatellite function and disease susceptibility. This selection bias was not a product of microsatellite instability in tumor samples, nor was there a correlation between NR0B1 GGAA-microsatellite polymorphisms and survival outcomes. CONCLUSIONS: These data suggest that GGAA-microsatellite polymorphisms observed in human populations modulate EWS/FLI-mediated gene expression and may influence disease susceptibility in Ewing sarcoma.

Academic affiliated training centers in humanitarian health, Part I: program characteristics and professionalization preferences of centers in North America.

The collaborative London based non-governmental organization network ELRHA (Enhancing Learning and Research for Humanitarian Assistance) supports partnerships between higher education institutions and humanitarian organizations worldwide with the objective to enhance the professionalization of the humanitarian sector. While coordination and control of the humanitarian sector has plagued the response to every major crisis, concerns highlighted by the 2010 Haitian earthquake response further catalyzed and accelerated the need to ensure competency-based professionalization of the humanitarian health care work force. The Harvard Humanitarian Initiative sponsored an independent survey of established academically affiliated training centers in North America that train humanitarian health care workers to determine their individual training center characteristics and preferences in the potential professionalization process. The survey revealed that a common thread of profession-specific skills and core humanitarian competencies were being offered in both residential and online programs with additional programs offering opportunities for field simulation experiences and more advanced degree programs. This study supports the potential for the development of like-minded academic affiliated and competency-based humanitarian health programs to organize themselves under ELRHA's regional "consultation hubs" worldwide that can assist and advocate for improved education and training opportunities in less served developing countries.

Panic attack history and anxiety sensitivity in relation to cognitive-based smoking processes among treatment-seeking daily smokers.

INTRODUCTION: Empirical research has found that panic attacks are related to increased risk of more severe nicotine withdrawal and poor cessation outcome. Anxiety sensitivity (AS; fear of anxiety and related sensations) has similarly been found to be related to an increased risk of acute nicotine withdrawal and poorer cessation outcome. However, research has yet to examine the relative contributions of panic attacks and AS in terms of cognitive-based smoking processes (e.g., negative reinforcement smoking expectancies, addictive and negative affect-based reduction smoking motives, barriers to cessation, problem symptoms experienced while quitting). METHOD: Participants (n = 242; 57.4% male; M (age) = 38.1) were daily smokers recruited as a part of a larger randomized control trial for smoking cessation. It was hypothesized that both panic attacks and AS would uniquely and independently predict the studied cognitive-based smoking processes. RESULTS: As hypothesized, AS was uniquely and positively associated with all smoking processes after controlling for average number of cigarettes smoked per day, current Axis I diagnosis, and participant sex. However, panic attack history was only significantly related to problem symptoms experienced while quitting smoking. CONCLUSIONS: Although past research has demonstrated significant associations between panic attacks and certain aspects of cigarette smoking (e.g., severity of nicotine withdrawal; lower abstinence rates, and negative affect reduction motives), the present findings suggest that AS may be more relevant to understanding beliefs about and motives for smoking behavior as well as perceptions of cessation-related difficulties.

Anxiety sensitivity and cognitive-based smoking processes: testing the mediating role of emotion dysregulation among treatment-seeking daily smokers.

The current study investigated whether emotion dysregulation (difficulties in the self-regulation of affective states) mediated relationships between anxiety sensitivity (fear of anxiety and related sensations) and cognitive-based smoking processes. Participants (n = 197; 57.5% male; mean age = 38.0 years) were daily smokers recruited as part of a randomized control trial for smoking cessation. Anxiety sensitivity was uniquely associated with all smoking processes. Moreover, emotion dysregulation significantly mediated relationships between anxiety sensitivity and the smoking processes. Findings suggest that emotion dysregulation is an important construct to consider in relationships between anxiety sensitivity and cognitive-based smoking processes among adult treatment-seeking smokers.

Main and interactive effects of emotion dysregulation and breath-holding duration in relation to panic-relevant fear and expectancies about anxiety-related sensations among adult daily smokers.

The current study investigated the main and interactive effects of emotion dysregulation and distress tolerance in relation to panic-relevant variables among daily smokers. The sample consisted of 172 adults (61.2% male; M(age)=31.58, SD=11.51), who reported smoking an average of 15.99 cigarettes per day (SD=10.00). Results indicated that both emotion dysregulation and distress tolerance were significantly related to interoceptive fear and agoraphobia. Additionally, emotion dysregulation, but not distress tolerance, was significantly related to anxiety sensitivity. All effects were evident above and beyond the variance accounted for by average cigarettes per day, tobacco-related physical illness, and panic attack history. The interaction between emotion dysregulation and distress tolerance significantly predicted interoceptive and agoraphobic fears as well as the cognitive component of anxiety sensitivity. Such findings underscore the importance of emotion dysregulation and distress tolerance in regard to panic-specific fear and expectancies about anxiety-related sensations among daily smokers.

Prospective evaluation of the effects of anxiety sensitivity and state anxiety in predicting acute nicotine withdrawal symptoms during smoking cessation.

The current investigation explored the main and interactive effects of anxiety sensitivity (AS) and state anxiety in predicting acute nicotine withdrawal symptoms experienced during the initial 14 days of smoking cessation. Participants included 123 adult daily smokers (84 women; Mage = 45.93 years, SD = 10.34) undergoing psychosocial-pharmacological cessation treatment. Results indicated that after controlling for the effects of participant sex and nicotine dependence, state anxiety but not AS significantly predicted initial levels of nicotine withdrawal symptoms. Results also demonstrated that both state anxiety and AS were significantly related to the change in nicotine withdrawal symptoms over time. Finally, our results revealed a significant interaction between AS and state anxiety. Specifically, higher levels of AS were associated with a stronger relation between state anxiety and nicotine withdrawal symptoms experienced during the cessation attempt. Results suggest that among high AS persons, state anxiety may be more relevant, compared to those low in AS, in regard to experiencing withdrawal symptoms as more intense during the early phases of quitting.

Microsatellites with macro-influence in ewing sarcoma.

Numerous molecular abnormalities contribute to the genetic derangements involved in tumorigenesis. Chromosomal translocations are a frequent source of these derangements, producing unique fusion proteins with novel oncogenic properties. EWS/ETS fusions in Ewing sarcoma are a prime example of this, resulting in potent chimeric oncoproteins with novel biological properties and a unique transcriptional signature essential for oncogenesis. Recent evidence demonstrates that EWS/FLI, the most common EWS/ETS fusion in Ewing sarcoma, upregulates gene expression using a GGAA microsatellite response element dispersed throughout the human genome. These GGAA microsatellites function as enhancer elements, are sites of epigenetic regulation and are necessary for EWS/FLI DNA binding and upregulation of principal oncogenic targets. An increasing number of GGAA motifs appear to substantially enhance EWS/FLI-mediated gene expression, which has compelling biological implications as these GGAA microsatellites are highly polymorphic within and between ethnically distinct populations. Historically regarded as junk DNA, this emerging evidence clearly demonstrates that microsatellite DNA plays an instrumental role in EWS/FLI-mediated transcriptional regulation and oncogenesis in Ewing sarcoma. This unprecedented role of GGAA microsatellite DNA in Ewing sarcoma provides a unique opportunity to expand our mechanistic understanding of how EWS/ETS fusions influence cancer susceptibility, prognosis and transcriptional regulation.

Chronic pain and marijuana use among a nationally representative sample of adults.

This study sought to examine the relations between chronic pain and marijuana use in a large nationally representative survey of adults (n = 5,672; 53% female; M(age) = 45.05, SD = 17.9) conducted in the United States. After controlling for sociodemographic variables, lifetime history of depression, and alcohol abuse/dependence, there was a significant association between lifetime chronic pain and lifetime and current marijuana use. Moreover, current chronic pain was significantly associated with lifetime marijuana use. There was no significant association between current chronic pain and current marijuana use, possibly owing to limited statistical power. Results suggest that there are generally consistent statistically significant relations between chronic pain and marijuana use. Future work is needed to explicate the developmental patterning between chronic pain and marijuana use. This paper presents the potential linkage between chronic pain and marijuana use. Results from this study suggest that it may be beneficial for clinicians to assess for marijuana use among patients suffering from chronic pain. Such patients may be using marijuana as a maladaptive coping strategy.

Personality disorders and cigarette smoking among adults in the United States.

INTRODUCTION: There is a paucity of empirical information pertaining to the association between personality disorders and cigarette smoking. The present study examined whether, and to what degree, personality disorders are associated with cigarette smoking; investigated the specificity of any observed smoking-personality disorder association; and the role of mood/anxiety disorders, substance use, and nicotine dependence in those relations. METHODS: Data were drawn from the National Epidemiologic Survey of Alcohol and Related Conditions (NESARC), a nationally representative sample of 43,083 adults in the United States. RESULTS: Results indicated a substantial percentage of those with personality disorders are nicotine dependent. Interestingly, the association between dependent, avoidant, histrionic, schizoid and paranoid personality disorders as well as former dependent smoking was partially explained by co-occurring mood/anxiety disorders, and adjusting for such clinical conditions appeared to generally attenuate the strength of many other associations. Finally, the association between personality disorders and smoking appears to differ by specific personality disorder, with some of the strongest relations being evident for antisocial personality disorder. DISCUSSION: These novel empirical findings are discussed in relation to the relevance of cigarette smoking among those with personality disorders.

Exploring the mediational role of coping motives for marijuana use in terms of the relation between anxiety sensitivity and marijuana dependence.

This study evaluated the prediction that coping motives for marijuana use would mediate the relation between anxiety sensitivity and a marijuana dependence diagnosis after controlling for other co-occurring marijuana use motives. Participants were 136 current marijuana users (47.1% women; M(age)= 21.9, SD = 7.2). Results were consistent with a mediational effect, with the relation between anxiety sensitivity and marijuana dependence being explained by the addition of coping motives into the model. These results provide novel information related to the putative explanatory role of coping motives for marijuana use in the relation between anxiety sensitivity and marijuana dependence.

Uni-morbid and co-occurring marijuana and tobacco use: examination of concurrent associations with negative mood states.

The aim of the current investigation was to examine uni-morbid and co-occurring tobacco and marijuana use in relation to the negative emotional symptoms of anxiety and depression. Participants were 250 adult individuals (132 women; mean age = 22.43 years, standard deviation = 9.00 years) who were divided into one of four non-overlapping substance use categories: tobacco use only (n = 39), marijuana use only (n = 62), co-occurring tobacco and marijuana use (n = 82), and neither tobacco nor marijuana use (n = 67). Results revealed three key findings. First, tobacco-only using individuals reported significantly greater negative affectivity than any of the other groups. Second, tobacco-only users reported greater anxious arousal symptoms than either the marijuana or non-substance use groups, but not the combined group. Third, tobacco-only users reported greater levels of depressive symptoms than either marijuana users or non-substance users. These findings provide novel information about tobacco and marijuana use and how these variables relate to the experience of general and specific types of negative emotional symptoms.

Perceived physical health and heart-focused anxiety among daily adult cigarette smokers: associations with affect-relevant smoking motives and outcome expectancies.

The present investigation examined heart-focused anxiety and perceived physical health in terms of affect-relevant cigarette smoking motives and outcome expectancies. Participants were a community sample of 140 adult daily smokers (81 women, mean age = 29.60 years, SD = 11.98). In terms of smoking motives, both heart-focused anxiety and perceived physical health incrementally predicted smoking for negative affect reduction motives above and beyond relevant covariates (gender, weekly alcohol consumption, daily smoking rate). Yet heart-focused anxiety, but not perceived physical health, was incrementally predictive of habitual motives, relative to the same covariates. With regard to smoking outcome expectancies, heart-focused anxiety was incrementally predictive of negative reinforcement outcome expectancies, whereas perceived physical health was not. Alternatively, perceived physical health was incrementally predictive of negative personal consequence outcome expectancies, but heart-focused anxiety was not. Findings are discussed in relation to the role of perceived health vulnerabilities in clarifying affect-oriented smoking motives and expectancies.