Systematic review of conservation interventions to promote voluntary behavior change.

Understanding human behavior is vital to developing interventions that effectively lead to proenvironmental behavior change, whether the focus is at the individual or societal level. However, interventions in many fields have historically lacked robust forms of evaluation, which makes it hard to be confident that these conservation interventions have successfully helped protect the environment. We conducted a systematic review to assess how effective nonpecuniary and nonregulatory interventions have been in changing environmental behavior. We applied the Office of Health Assessment and Translation systematic review methodology. We started with more than 300,000 papers and reports returned by our search terms and after critical appraisal of quality identified 128 individual studies that merited inclusion in the review. We classified interventions by thematic area, type of intervention, the number of times audiences were exposed to interventions, and the length of time interventions ran. Most studies reported a positive effect (n = 96). The next most common outcome was no effect (n = 28). Few studies reported negative (n = 1) or mixed (n = 3) effects. Education, prompts, and feedback interventions resulted in positive behavior change. Combining multiple interventions was the most effective. Neither exposure duration nor frequency affected the likelihood of desired behavioral change. Comparatively few studies tested the effects of voluntary interventions on non-Western populations (n = 17) or measured actual ecological outcome behavior (n = 1). Similarly, few studies examined conservation devices (e.g., energy-efficient stoves) (n = 9) and demonstrations (e.g., modeling the desired behavior) (n = 5). There is a clear need to both improve the quality of the impact evaluation conducted and the reporting standards for intervention results.

Es vital entender el comportamiento humano para desarrollar intervenciones que lleven efectivamente a un cambio proambiental en el comportamiento, ya sea que estén enfocadas a nivel individual o de sociedad. Sin embargo, históricamente las intervenciones han carecido de evaluaciones sólidas en muchas áreas, lo que dificulta confiar en que estas intervenciones de conservación hayan ayudado exitosamente a proteger el ambiente. Realizamos una revisión sistemática para analizar cuán efectivas han sido las intervenciones no pecuniarias y no regulatorias en el cambio del comportamiento ambiental. Aplicamos la metodología de revisión sistemática de la Oficina de Evaluación y Traducción de la Salud (OETS). Comenzamos con más de 300,000 artículos y reportes arrojados por nuestros términos de búsqueda y después de una valoración crítica de calidad, identificamos 128 estudios individuales que merecían su inclusión dentro de la revisión. Clasificamos las intervenciones en área temática, tipo de intervención, número de veces que el público estuvo expuesto a las intervenciones y duración de las intervenciones. La mayoría de los estudios reportaron un efecto positivo (n = 96). El siguiente resultado más común fue sin efecto (n = 28). Pocos estudios reportaron efectos negativos (n = 1) o mixtos (n = 3). Las intervenciones educativas, de indicaciones y de retroalimentación resultaron en un cambio conductual positivo. La combinación de las múltiples intervenciones fue la más efectiva. Ni la duración de la exposición ni la frecuencia afectaron la probabilidad del cambio conductual deseado. En comparación, pocos estudios analizaron los efectos de las intervenciones voluntarias sobre las poblaciones no occidentales (n = 17) o midieron el comportamiento ecológico resultante actual (n = 1). De manera similar, pocos estudios examinaron los dispositivos (p. ej.: estufas ahorradoras de energía) (n = 9) y demostraciones (p. ej.: modelar un comportamiento deseado) (n = 5) de conservación. Hay una necesidad evidente de mejorar la calidad de las evaluaciones de impacto realizadas y de reportar los estándares para los resultados de las intervenciones.

Experimental evidence of the impact of framing of actors and victims in conservation narratives.

Media narratives play a crucial role in framing marine conservation dilemmas by depicting human actors, such as fish consumers or the fishing industry, as responsible for negative effects of their actions on species and ecosystems. However, there is little evidence documenting how such narratives affect preferences for reducing bycatch. Behavioral science research shows that people can act less prosocially when more actors are responsible for a collective outcome (responsibility diffusion effect) and when more victims need to be helped (compassion fade effect); thus, the media's framing of actors and victims may have a significant effect on preferences. We conducted the first test of responsibility diffusion and compassion fade in a marine context in an online experiment (1548 participants in the United Kingdom). In 9 media narratives, we varied the type of actors responsible for fisheries bycatch (e.g., consumers and industry) and victims (e.g., a single species, multiple species, and ecosystems) in media narratives and determined the effects of the narratives on participants' support for bycatch policies and intentions to alter fish consumption. When responsibility for negative effects was attributed to consumers and industry, the probability of participants reporting support for fisheries policies (e.g., bycatch enforcement or consumer taxes) was ∼30% higher (odds ratio = 1.32) than when only consumers were attributed responsibility. These effects were primarily driven by female participants. Narratives had no effect on personal intentions to consume fish. Varying the type of victim had no effect on policy support and intentions. Our results suggest that neither responsibility diffusion nor compassion fade automatically follows from increasing the types of actors and victims in media narratives and that effects can depend on the type of outcome and population subgroup.

Las narrativas mediáticas juegan un papel importante en el encuadre de los dilemas de conservación marina al representar a los actores humanos, por ejemplo, los consumidores de pescado o la industria pesquera, como los responsables de los efectos negativos de sus acciones sobre las especies y ecosistemas. Sin embargo, hay poca evidencia que documente cómo estas narrativas afectan las preferencias para reducir la captura incidental. Investigaciones de la ciencia conductual muestran que las personas pueden actuar menos a favor de la sociedad cuando más actores son responsables de un resultado colectivo (efecto de difusión de la responsabilidad) y cuando más víctimas necesitan asistencia (efecto de la desaparición de la compasión); por lo tanto, el encuadre mediático de los actores y las víctimas puede tener un efecto significativo sobre las preferencias. Realizamos el primer análisis de la difusión de la responsabilidad y la desaparición de la compasión en un contexto marino con un experimento en línea (1,548 participantes en el Reino Unido). Diversificamos el tipo de actores responsables de la captura incidental (p. ej.: consumidores e industria) y sus víctimas (p. ej.: una sola especie, múltiples especies y ecosistemas) en nueve narrativas mediáticas y determinamos sus efectos sobre el respaldo que dan los participantes a las políticas de captura incidental y sus intenciones de alterar el consumo de pescado. Cuando se le atribuyó la responsabilidad de los efectos negativos a los consumidores y a la industria, la probabilidad de que los participantes apoyaran las políticas pesqueras (p. ej.: implementación de la captura incidental o impuestos al consumidor) fue ∼30% más alta (razón de probabilidad = 1.32) que cuando se le atribuyó la responsabilidad solamente a los consumidores. Estos efectos fueron impulsados principalmente por las mujeres participantes. Las narrativas no tuvieron efectos sobre las intenciones personales de consumir pescado. La variación en el tipo de víctimas no tuvo efectos sobre el apoyo a las políticas y las intenciones. Nuestros resultados sugieren que ni la difusión de la responsabilidad ni la desaparición de la compasión ocurren automáticamente tras incrementar los tipos de actores y víctimas en las narrativas mediáticas y que los efectos pueden depender del tipo de resultado y del subgrupo poblacional. Evidencia Experimental del Impacto que Tiene el Encuadre de los Actores y las Víctimas en las Narrativas de Conservación.

Omission of Radiation in Conservative Treatment for Breast Cancer: Opportunity for De-escalation of Care.

INTRODUCTION: De-escalation of breast cancer treatment aims to reduce patient and financial toxicity without compromising outcomes. Level I evidence and National Comprehensive Cancer Network guidelines support omission of adjuvant radiation in patients aged >70 y with hormone-sensitive, pT1N0M0 invasive breast cancer treated with endocrine therapy. We evaluated radiation use in patients eligible for guideline concordant omission of radiation. METHODS: Subgroup analysis of patients eligible for radiation omission from two pooled randomized controlled trials, which included stage 0-III breast cancer patients undergoing breast conserving surgery, was performed to evaluate factors associated with radiation use. RESULTS: Of 631 patients, 47 (7.4%) met radiation omission criteria and were treated by 14 surgeons at eight institutions. The mean age was 75.3 (standard deviation + 4.4) y. Majority of patients identified as White (n = 46; 97.9%) and non-Hispanic (n = 44; 93.6%). The mean tumor size was 1.0 cm; 37 patients (88.1%) had ductal, 4 patients (9.5%) had lobular, and 17 patients (40.5%) had low-grade disease. Among patients eligible for radiation omission, 34 (72.3%) patients received adjuvant radiation. Those who received radiation were significantly younger than those who did not (74 y, interquartile range = 4 y, versus 78 y, interquartile range = 11 y, P = 0.03). There was no difference in radiation use based on size (P = 0.4), histology (P = 0.5), grade (P = 0.7), race (P = 1), ethnicity (P = 0.6), institution (P = 0.1), gender of the surgeon (P = 0.7), or surgeon (P = 0.1). CONCLUSIONS: Fewer than 10% of patients undergoing breast conservation met criteria for radiation omission. Nearly three-quarters received radiation therapy with younger age being a driver of radiation use, suggesting ample opportunity for de-escalation, particularly among younger eligible patients.

Are we choosing wisely? Drivers of preoperative MRI use in breast cancer patients.

INTRODUCTION: Factors contributing to the use of preoperative MRI remain poorly understood. METHODS: Data from a randomized controlled trial of stage 0-3 breast cancer patients undergoing breast conserving surgery between 2016 and 2018 were analyzed. RESULTS: Of the 396 patients in this trial, 32.6% had a preoperative MRI. Patient age, race, ethnicity, tumor histology, and use of neoadjuvant therapy were significant predictors of MRI use. On multivariate analysis, younger patients with invasive lobular tumors were more likely to have a preoperative MRI. Rates also varied significantly by individual surgeon (p < 0.001); in particular, female surgeons (39.9% vs. 24.0% for male surgeons, p = 0.001) and those in community practice (58.9% vs. 14.2% for academic, p < 0.001) were more likely to order preoperative MRI. Rates declined over the two years of the study, particularly among female surgeons. CONCLUSIONS: Preoperative MRI varies with patient age and tumor histology; however, there remains variability by individual surgeon.

Factors Affecting Time to Surgery in Breast Cancer Patients.

BACKGROUND: We sought to determine factors affecting time to surgery (TTS) to identify potential modifiable factors to improve timeliness of care. METHODS: Patients with clinical stage 0-3 breast cancer undergoing partial mastectomy in 2 clinical trials, conducted in ten centers across the US, were analyzed. No preoperative workup was mandated by the study; those receiving neoadjuvant therapy were excluded. RESULTS: The median TTS among the 583 patients in this cohort was 34 days (range: 1-289). Patient age, race, tumor palpability, and genomic subtype did not influence timeliness of care defined as TTS ≤30 days. Hispanic patients less likely to have a TTS ≤30 days (P = .001). There was significant variation in TTS by surgeon (P < .001); those practicing in an academic center more likely to have TTS ≤30 days than those in a community setting (55.1% vs 19.3%, P < .001). Patients who had a preoperative ultrasound had a similar TTS to those who did not (TTS ≤30 days 41.9% vs 51.9%, respectively, P = .109), but those who had a preoperative MRI had a significantly increased TTS (TTS ≤30 days 25.0% vs 50.9%, P < .001). On multivariate analysis, patient ethnicity was no longer significantly associated with TTS ≤30 (P = .150). Rather, use of MRI (OR: .438; 95% CI: .287-.668, P < .001) and community practice type (OR: .324; 95% CI: .194-.541, P < .001) remained independent predictors of lower likelihood of TTS ≤30 days. CONCLUSIONS: Preoperative MRI significantly increases time to surgery; surgeons should consider this in deciding on its use.

Resection of Cavity Shave Margins in Stage 0-III Breast Cancer Patients Undergoing Breast Conserving Surgery: A Prospective Multicenter Randomized Controlled Trial.

OBJECTIVE: Single-center studies have demonstrated that resection of cavity shave margins (CSM) halves the rate of positive margins and re-excision in breast cancer patients undergoing partial mastectomy (PM). We sought to determine if these findings were externally generalizable across practice settings. METHODS: In this multicenter randomized controlled trial occurring in 9 centers across the United States, stage 0-III breast cancer patients undergoing PM were randomly assigned to either have resection of CSM ("shave" group) or not ("no shave" group). Randomization occurred intraoperatively, after the surgeon had completed their standard PM. Primary outcome measures were positive margin and re-excision rates. RESULTS: Between July 28, 2016 and April 13, 2018, 400 patients were enrolled in this trial. Four patients (2 in each arm) did not meet inclusion criteria after randomization, leaving 396 patients for analysis: 196 in the "shave" group and 200 to the "no shave" group. Median patient age was 65 years (range; 29-94). Groups were well matched at baseline for demographic and clinicopathologic factors. Prior to randomization, positive margin rates were similar in the "shave" and "no shave" groups (76/196 (38.8%) vs. 72/200 (36.0%), respectively, P = 0.604). After randomization, those in the "shave" group were significantly less likely than those in the "no shave" group to have positive margins (19/196 (9.7%) vs. 72/200 (36.0%), P < 0.001), and to require re-excision or mastectomy for margin clearance (17/196 (8.7%) vs. 47/200 (23.5%), P < 0.001). CONCLUSION: Resection of CSM significantly reduces positive margin and re-excision rates in patients undergoing PM.

Motivations for the use and consumption of wildlife products.

The dominant approach to combating the illegal wildlife trade has traditionally been to restrict the supply of wildlife products. Yet conservationists increasingly recognize the importance of implementing demand-side interventions that target the end consumers in the trade chain. Their aim is to curb the consumption of wildlife or shift consumption to more sustainable alternatives. However, there are still considerable knowledge gaps in understanding of the diversity of consumer motivations in the context of illegal wildlife trade, which includes hundreds of thousands of species, different uses, and diverse contexts. Based on consultation with multiple experts from a diversity of backgrounds, nationalities, and focal taxa, we developed a typology of common motivations held by wildlife consumers that can be used to inform conservation interventions. We identified 5 main motivational categories for wildlife use: experiential, social, functional, financial, and spiritual, each containing subcategories. This framework is intended to facilitate the segmentation of consumers based on psychographics and allow the tailoring of interventions-whether behavior change campaigns, enforcement efforts, or incentive programs-to the specific context in which they will be used. Underlining the importance of consumer research and collaborating with local actors is an important step toward promoting a more systematic approach to the design of demand reduction interventions.

Motivaciones para el Uso y Consumo de Productos de Fauna Resumen Tradicionalmente, la estrategia dominante para combatir el mercado ilegal de fauna ha sido restringir la oferta de productos de fauna. Aun así, los conservacionistas cada vez reconocen más la importancia de implementar intervenciones por el lado de la demanda que se enfoquen en los consumidores finales en la cadena de mercado. Su objetivo es reducir el consumo de fauna o redirigir ese consumo hacia alternativas más sustentables. Sin embargo, todavía existen vacíos de conocimiento en el entendimiento de la diversidad de motivos para los consumidores dentro del contexto del mercado ilegal de fauna, el cual incluye cientos de miles de especies, diferentes usos y contextos diversos. Con base en consultas a varios expertos con una diversidad de antecedentes, nacionalidades y taxones de enfoque desarrollamos una tipología de motivos comunes que tienen los consumidores de fauna que pueden usarse para orientar las intervenciones de conservación. Identificamos cinco categorías principales de motivos para el uso de fauna: vivencial, social, funcional, financiero y espiritual, cada uno con subcategorías. Este marco de trabajo tiene la intención de facilitar la segmentación de consumidores con base en psicográficos y permitir la personalización de las intervenciones - sean campañas de cambios en el comportamiento, esfuerzos por hacer cumplir las reglas o programas de incentivos - al contexto específico en el que serán utilizadas. Resaltar la importancia de los estudios de mercado y la colaboración con los actores locales es un paso importante hacia la promoción de una estrategia más sistemática para el diseño de intervenciones para reducir la demanda.

Cystocerebral Syndrome: An Updated Review and a New Proposed Mechanism for an Often Forgotten Cause of Delirium.

Cystocerebral syndrome is an often forgotten cause of delirium in elderly males, which is quite easily treated. We reviewed the current body of literature documenting cystocerebral syndrome and proposed a new mechanism of action explaining why all patients identified thus far have been male. Data was obtained from articles describing cases of cystocerebral syndrome, urinary retention, and confusion in addition to delirium via a PubMed database search. We reviewed all articles describing cases of cystocerebral syndrome via the PubMed database using the Medical Subject Headings (MeSH) keywords of "cystocerebral syndrome," urinary retention and confusion," and "delirium and urinary retention or cystocerebral syndrome," and identified eight cases of cystocerebral syndrome including the original publication by Blackburn and Dunn. We found that all patients reported in the literature were males older than 70 years and often with concomitant benign prostatic hypertrophy (BPH) who presented with acute episodes of delirium that rapidly responded to bladder decompression. The authors seek to update the medical community regarding this uncommon phenomenon of delirium in elderly male patients. We also propose that the lack of female patients in the literature is reflective of their decreased intraurethral flow resistance as is currently being described in other avenues of research in the field of urodynamics.

Experimental Therapeutics for Challenging Clinical Care of a Patient with an Extremely Rare Homozygous APOC2 Mutation.

BACKGROUND: Among many causes of hypertriglyceridemia (HTG), familial chylomicronemia syndrome (FCS) is a rare monogenic disorder that manifests as severe HTG and acute pancreatitis. Among the known causal genes for FCS, mutations in APOC2 only account for <2% of cases. Medical nutrition therapy is critical for FCS because usual triglyceride- (TG-) lowering medications are ineffective. Therapeutic plasma exchange (TPE) with fresh frozen plasma (FFP) is an option to urgently reduce TG and pancreatitis episodes. Several novel biologics are under development to treat HTG and may provide therapeutic options for FCS in the future. OBJECTIVE: We present the challenging care of a 43-year-old man with FCS with apoC-II deficiency and the results of two types of TPE and of investigational TG-lowering biologic therapies. RESULTS: The patient's lipid profile was consistent with FCS. A novel homozygous variant was identified in APOC2, and its pathogenicity was confirmed. Even on a fat-restricted diet, his care was tremendously complicated with unremitting bouts of pancreatitis. TPE with FFP replacement lowered TG >90% post-sessions and appeared to reduce pancreatitis episodes. Experimental ANGPTL3 and APOC3 inhibitors each lowered TG by >50%. CONCLUSIONS: Our case demonstrates the importance of delineating and defining the underlying etiology of a rare disorder to optimize therapy and to minimize unfavorable outcomes.

A scoping review into the impact of animal imagery on pro-environmental outcomes.

With the recognition that most global environmental problems are a result of human actions, there is an increasing interest in approaches which have the potential to influence human behaviour. Images have a powerful role in shaping persuasive messages, yet research on the impacts of visual representations of nature is a neglected area in biodiversity conservation. We systematically screened existing studies on the use of animal imagery in conservation, identifying 37 articles. Although there is clear evidence that images of animals can have positive effects on people's attitudes to animals, overall there is currently a dearth of accessible and comparable published data demonstrating the efficacy of animal imagery. Most existing studies are place and context-specific, limiting the generalisable conclusions that can be drawn. Transdisciplinary research is needed to develop a robust understanding of the contextual and cultural factors that affect how animal images can be used effectively for conservation purposes.

EUS-guided fine-needle core liver biopsy sampling using a novel 19-gauge needle with modified 1-pass, 1 actuation wet suction technique.

BACKGROUND AND AIMS: EUS-guided fine-needle core biopsy sampling is a safe and effective technique for diagnosis of focal liver lesions. However, data are limited in its role in parenchymal disease. We evaluated the utility of EUS-guided parenchymal liver biopsy sampling with a modified 1-pass wet suction technique (EUS-modified liver biopsy sampling [EUS-MLB]) in patients with unexplained increase in liver-associated tests. METHODS: We retrospectively evaluated the safety and efficacy of EUS-MLB in patients referred for EUS to evaluate for biliary obstruction and pancreatic disorders but with associated unexplained liver tests. EUS-MLB was performed during the same session after biliary obstruction was excluded. RESULTS: One hundred sixty-five consecutive patients underwent EUS-MLB. The median age was 52 years (interquartile range [IQR], 42-65). Sixty-eight patients (41%) were men. The median of the maximum intact core tissue length was 2.4 cm (IQR, 1.8-3.5). The median total specimen length (TSL) was 6 cm (IQR, 4.3-8). The median number of complete portal tracts (CPTs) per TSL was 18 (IQR, 13- 24). The mean number of CPTs per sample length was 7.5 cm. Adverse events were uncommon (1.8%) and included abdominal pain and self-limited hematoma. CONCLUSIONS: EUS-guided fine-needle biopsy sampling using a novel 19-gauge core needle with a modified 1-pass 1 actuation wet suction technique (EUS-MLB) is a safe and effective way to evaluate patients with unexplained liver tests abnormalities who are undergoing EUS for exclusion of biliary obstruction.

A quantitative-PCR based method to estimate ranavirus viral load following normalisation by reference to an ultraconserved vertebrate target.

Ranaviruses are important pathogens of amphibians, reptiles and fish. To meet the need for an analytical method for generating normalised and comparable infection data for these diverse host species, two standard-curve based quantitative-PCR (qPCR) assays were developed enabling viral load estimation across these host groups. A viral qPCR targeting the major capsid protein (MCP) gene was developed which was specific to amphibian-associated ranaviruses with high analytical sensitivity (lower limit of detection: 4.23 plasmid standard copies per reaction) and high reproducibility across a wide dynamic range (coefficient of variation below 3.82% from 3 to 3×108 standard copies per reaction). The comparative sensitivity of the viral qPCR was 100% (n=78) based on agreement with an established end-point PCR. Comparative specificity with the end-point PCR was also 100% (n=94) using samples from sites with no history of ranavirus infection. To normalise viral quantities, a host qPCR was developed which targeted a single-copy, ultra-conserved non-coding element (UCNE) of vertebrates. Viral and host qPCRs were applied to track ranavirus growth in culture. The two assays offer a robust approach to viral load estimation and the host qPCR can be paired with assays targeting other pathogens to study infection burdens.

Minireview: Role of genetic changes of faciogenital dysplasia protein 1 in human disease.

The FGD1 gene encodes for a guanine exchange factor (GEF) protein that specifically activates the Rho GTPase Cdc42. For cellular migration, Cdc42 is a key molecular switch that regulates cytoskeleton restructuring, gene transcription, cellular morphology, extension, and cell adhesion. In the past decade, germline mutations in the FGD1 gene have been associated with a rare X-linked disorder known as faciogenital dysplasia (FGDY). Malformations are consistent with a loss of cellular migration during embryonic development. Insertion and deletion mutations in FGD1 result in a frameshift causing inactivation of fgd1 protein. Since Cdc42 is a key molecular switch in cytoskeletal restructuring and cell adhesion, the loss of fgd1 is postulated to attenuate Cdc42-mediated cellular migration in embryonic development. In metastatic tumors, Cdc42 modulates migration and invasiveness. Fgd1 overexpression has been found in infiltrating and poorly differentiated breast and invasive prostate tumors. Amplification at Xp11.21, the FGD1 gene locus, has been reported in several cancers. Sequencing analyses in numerous types of cancer have found missense mutations in the FGD1 gene in metastatic tumors. FGDY and cancer studies suggest that the germline and somatic changes downregulate or upregulate the FGD1 gene playing a key role in the development of diseases.

The AtGRXS3/4/5/7/8 glutaredoxin gene cluster on Arabidopsis thaliana chromosome 4 is coordinately regulated by nitrate and appears to control primary root growth.

Nitrate and ammonium are the 2 most common forms of inorganic nitrogen available to plants in the soil. We previously identified a group of class III glutaredoxin genes whose expression is strongly upregulated by nitrate, but not ammonium, in Arabidopsis thaliana shoots and roots. A reverse genetics approach was used to functionally characterize a subset of these nitrate-regulated glutaredoxins, and we found that the AtGRXS3,4,5, and 8 genes function as negative regulators of primary root growth. AtGRXS3/4/5/8 are arranged in a tandem array on Arabidopsis chromosome 4, and these genes show very high levels of sequence similarity. Interestingly, there is one additional glutaredoxin, AtGRXS7, in this same gene cluster, but this gene was not identified as nitrate-responsive in our previous studies. We show here that AtGRXS7 is upregulated by nitrate and shows strong co-expression with the other glutaredoxins in this gene cluster. Further, AtGRXS7 was effectively silenced by the RNAi construct used to target AtGRXS3/4/5/8 for previous functional analyses. Overall, it appears that the 5 genes in the AtGRX3/4/5/7/8 cluster share virtually identical sequences, regulatory patterns, and functions, collectively acting to regulate primary root growth in response to soil nitrate.

Nitrate-Regulated Glutaredoxins Control Arabidopsis Primary Root Growth.

Nitrogen is an essential soil nutrient for plants, and lack of nitrogen commonly limits plant growth. Soil nitrogen is typically available to plants in two inorganic forms: nitrate and ammonium. To better understand how nitrate and ammonium differentially affect plant metabolism and development, we performed transcriptional profiling of the shoots of ammonium-supplied and nitrate-supplied Arabidopsis (Arabidopsis thaliana) plants. Seven genes encoding class III glutaredoxins were found to be strongly and specifically induced by nitrate. RNA silencing of four of these glutaredoxin genes (AtGRXS3/4/5/8) resulted in plants with increased primary root length (approximately 25% longer than the wild type) and decreased sensitivity to nitrate-mediated inhibition of primary root growth. Increased primary root growth is also a well-characterized phenotype of many cytokinin-deficient plant lines. We determined that nitrate induction of glutaredoxin gene expression was dependent upon cytokinin signaling and that cytokinins could activate glutaredoxin gene expression independent of plant nitrate status. In addition, crosses between "long-root" cytokinin-deficient plants and "long-root" glutaredoxin-silenced plants generated hybrids that displayed no further increase in primary root length (i.e. epistasis). Collectively, these findings suggest that AtGRXS3/4/5/8 operate downstream of cytokinins in a signal transduction pathway that negatively regulates plant primary root growth in response to nitrate. This pathway could allow Arabidopsis to actively discriminate between different nitrogen sources in the soil, with the preferred nitrogen source, nitrate, acting to suppress primary root growth (vertical dimension) in concert with its well-characterized stimulatory effect on lateral root growth (horizontal dimension).

Characterization and pharmacologic targeting of EZH2, a fetal retinal protein and epigenetic regulator, in human retinoblastoma.

Retinoblastoma (RB) is the most common primary intraocular cancer in children, and the third most common cancer overall in infants. No molecular-targeted therapy for this lethal tumor exists. Since the tumor suppressor RB1, whose genetic inactivation underlies RB, is upstream of the epigenetic regulator EZH2, a pharmacologic target for many solid tumors, we reasoned that EZH2 might regulate human RB tumorigenesis. Histologic and immunohistochemical analyses were performed using an EZH2 antibody in sections from 43 samples of primary, formalin-fixed, paraffin-embedded human RB tissue, cryopreserved mouse retina, and in whole cell lysates from human RB cell lines (Y79 and WERI-Rb1), primary human fetal retinal pigment epithelium (RPE) and fetal and adult retina, mouse retina and embryonic stem (ES) cells. Although enriched during fetal human retinal development, EZH2 protein was not present in the normal postnatal retina. However, EZH2 was detected in all 43 analyzed human RB specimens, indicating that EZH2 is a fetal protein expressed in postnatal human RB. EZH2 expression marked single RB cell invasion into the optic nerve, a site of invasion whose involvement may influence the decision for systemic chemotherapy. To assess the role of EZH2 in RB cell survival, human RB and primary RPE cells were treated with two EZH2 inhibitors (EZH2i), GSK126 and SAH-EZH2 (SAH). EZH2i impaired intracellular adenosine triphosphate (ATP) production, an indicator of cell viability, in a time and dose-dependent manner, but did not affect primary human fetal RPE. Thus, aberrant expression of a histone methyltransferase protein is a feature of human RB. This is the first time this mechanism has been implicated for an eye, adnexal, or orbital tumor. The specificity of EZH2i toward human RB cells, but not RPE, warrants further in vivo testing in animal models of RB, especially those EZH2i currently in clinical trials for solid tumors and lymphoma.

Ductal carcinoma in situ with distorting sclerosis on core biopsy may be predictive of upstaging on excision.

AIMS: The aim of this study was to examine clinicopathological features of patients with core biopsy diagnoses of ductal carcinoma in situ (DCIS) that may predict invasion on subsequent excision, as upstaging has significant implications regarding the need for axillary staging via sentinel lymph node biopsy (SLNB). METHODS AND RESULTS: We identified 186 patients with a diagnosis of DCIS as the highest-stage lesion on core biopsy. Pathological and clinical features were assessed via slide and chart review, respectively. Distorting sclerosis was defined as irregular angulation of glands involved by DCIS but lacking definite invasion according to histology and/or immunohistochemical staining for myoepithelial markers. Thirty-two of 186 (17.2%) cases had upstaging to either microinvasive (nine) or invasive (23) ductal carcinoma. SLNB was performed in 29 of 32 (90.6%) cases with upstaging and in 55 of 154 (35.7%) cases without (P < 0.0001). Upstaging was significantly associated with the presurgical variables of radiological mass (P = 0.009) and distorting sclerosis (P = 0.0005) and the postsurgical feature of multifocality (P < 0.0001). CONCLUSIONS: Sentinel lymph node biopsy is frequently performed for patients with upstaging from DCIS on core biopsy to microinvasive or invasive carcinoma on excision. DCIS with distorting sclerosis without definite invasion on core biopsy may be predictive of upstaging. This feature may be useful in selecting patients to undergo SLNB at the time of excision to avoid reoperation.

Effectiveness of methylcobalamin and folinic Acid treatment on adaptive behavior in children with autistic disorder is related to glutathione redox status.

Treatments targeting metabolic abnormalities in children with autism are limited. Previously we reported that a nutritional treatment significantly improved glutathione metabolism in children with autistic disorder. In this study we evaluated changes in adaptive behaviors in this cohort and determined whether such changes are related to changes in glutathione metabolism. Thirty-seven children diagnosed with autistic disorder and abnormal glutathione and methylation metabolism were treated with twice weekly 75 µg/Kg methylcobalamin and twice daily 400 µg folinic acid for 3 months in an open-label fashion. The Vineland Adaptive Behavior Scale (VABS) and glutathione redox metabolites were measured at baseline and at the end of the treatment period. Over the treatment period, all VABS subscales significantly improved with an average effect size of 0.59, and an average improvement in skills of 7.7 months. A greater improvement in glutathione redox status was associated with a greater improvement in expressive communication, personal and domestic daily living skills, and interpersonal, play-leisure, and coping social skills. Age, gender, and history of regression did not influence treatment response. The significant behavioral improvements observed and the relationship between these improvements to glutathione redox status suggest that nutritional interventions targeting redox metabolism may benefit some children with autism.

DEK expression in Merkel cell carcinoma and small cell carcinoma.

BACKGROUND: The chromatin architectural factor DEK maps to chromosome 6p and is frequently overexpressed in several neoplasms, including small cell lung carcinoma, where it is associated with poor prognosis, tumor initiation activity and chemoresistance. DEK expression has not been studied in cutaneous Merkel cell carcinoma. METHODS: We applied a DEK monoclonal antibody to 15 cases of Merkel cell carcinoma and 12 cases of small cell carcinoma. DEK nuclear immunoreactivity was scored based on percentage (0, negative; 1+, <25%; 2+, 25-50%; 3+, >50%) and intensity (weak, moderate or strong). RESULTS: All 15 Merkel cell carcinoma cases (100%) showed diffuse (3+) nuclear positivity (14 strong, 1 weak). Six of 12 small cell carcinoma cases (50%) showed diffuse (3+) and strong nuclear positivity, while one case exhibited focal (1+) weak nuclear positivity. The remaining five cases were negative for DEK expression. CONCLUSIONS: Our results suggest that DEK may be involved in the pathogenesis of Merkel cell carcinoma and therefore may provide therapeutic implications for Merkel cell carcinomas. In addition, the difference in DEK expression between Merkel cell carcinoma and small cell carcinoma suggests possible separate tumorigenesis pathways for the two tumors.