Tumour budding as a prognostic biomarker in biopsies and resections of neoadjuvant-treated rectal adenocarcinoma.

BACKGROUND: Tumour budding (TB) is a marker of tumour aggressiveness which, when measured in rectal cancer resection specimens, predicts worse outcomes and response to neoadjuvant therapy. We investigated the utility of TB assessment in the setting of neoadjuvant treatment. METHODS AND RESULTS: A single-centre, retrospective cohort study was conducted. TB was assessed using the hot-spot International Tumour Budding Consortium (ITBCC) method and classified by the revised ITBCC criteria. Haematoxylin and eosin (H&E) and AE1/AE3 cytokeratin (CK) stains for ITB (intratumoural budding) in biopsies with PTB (peritumoural budding) and ITB (intratumoural budding) in resection specimens were compared. Logistic regression assessed budding as predictors of lymph node metastasis (LNM). Cox regression and Kaplan-Meier analyses investigated their utility as a predictor of disease-free (DFS) and overall (OS) survival. A total of 146 patients were included; 91 were male (62.3%). Thirty-seven cases (25.3%) had ITB on H&E and 79 (54.1%) had ITB on CK assessment of biopsy tissue. In univariable analysis, H&E ITB [odds (OR) = 2.709, 95% confidence interval (CI) = 1.261-5.822, P = 0.011] and CK ITB (OR = 2.165, 95% CI = 1.076-4.357, P = 0.030) predicted LNM. Biopsy-assessed H&E ITB (OR = 2.749, 95% CI = 1.258-6.528, P = 0.022) was an independent predictor of LNM. In Kaplan-Meier analysis, ITB identified on biopsy was associated with worse OS (H&E, P = 0.003, CK: P = 0.009) and DFS (H&E, P = 0.012; CK, P = 0.045). In resection specimens, CK PTB was associated with worse OS (P = 0.047), and both CK PTB and ITB with worse DFS (PTB, P = 0.014; ITB: P = 0.019). In multivariable analysis H&E ITB predicted OS (HR = 2.930, 95% CI = 1.261-6.809) and DFS (HR = 2.072, 95% CI = 1.031-4.164). CK PTB grading on resection also independently predicted OS (HR = 3.417, 95% CI = 1.45-8.053, P = 0.005). CONCLUSION: Assessment of TB using H&E and CK may be feasible in rectal cancer biopsy and post-neoadjuvant therapy-treated resection specimens and is associated with LNM and worse survival outcomes. Future management strategies for rectal cancer might be tailored to incorporate these findings.

Introducing a common taxonomy to support learning from failure in conservation.

Although some sectors have made significant progress in learning from failure, there is currently limited consensus on how a similar transition could best be achieved in conservation and what is required to facilitate this. One of the key enabling conditions for other sectors is a widely accepted and standardized classification system for identifying and analyzing root causes of failure. We devised a comprehensive taxonomy of root causes of failure affecting conservation projects. To develop this, we solicited examples of real-life conservation efforts that were deemed to have failed in some way, identified their underlying root causes of failure, and used these to develop a generic, 3-tier taxonomy of the ways in which projects fail, at the top of which are 6 overarching cause categories that are further divided into midlevel cause categories and specific root causes. We tested the taxonomy by asking conservation practitioners to use it to classify the causes of failure for conservation efforts they had been involved in. No significant gaps or redundancies were identified during this testing phase. We then analyzed the frequency that particular root causes were encountered by projects within this test sample, which suggested that some root causes were more likely to be encountered than others and that a small number of root causes were more likely to be encountered by projects implementing particular types of conservation action. Our taxonomy could be used to improve identification, analysis, and subsequent learning from failed conservation efforts, address some of the barriers that currently limit the ability of conservation practitioners to learn from failure, and contribute to establishing an effective culture of learning from failure within conservation.

Introducción de una taxonomía común como apoyo al aprendizaje a partir del fracaso en la conservación Resumen Mientras que algunos sectores han progresado significativamente en el aprendizaje a partir del fracaso, actualmente hay un consenso limitado sobre cómo podría lograrse una transición similar en la conservación y qué se requiere para facilitarla. Una de las condiciones habilitantes más importantes en otros sectores es un sistema de clasificación estandarizado y aceptado por la mayoría para la identificación y análisis de las causas fundamentales del fracaso. Diseñamos una taxonomía completa de las causas fundamentales del fracaso que afecta a los proyectos de conservación. Para desarrollarla, solicitamos ejemplos de esfuerzos de conservación reales que de alguna manera se consideraron como fracasos, identificamos las causas fundamentales no aparentes de su fracaso y luego las usamos para desarrollar una taxonomía genérica de tres niveles de las maneras en las que fracasan los proyectos, en cuyo nivel superior están seis categorías de causas generales que después se dividen en categorías de nivel medio de categorías de causas y causas fundamentales específicas. Pusimos a prueba la taxonomía al pedirle a los practicantes de la conservación que la usaran para clasificar las causas del fracaso de los esfuerzos de conservación en los que han participado. No identificamos vacíos o redundancias importantes durante esta fase de prueba. Después, analizamos la frecuencia con la que los proyectos de esta muestra se enfrentaron a causas fundamentales particulares, lo que sugirió que algunas causas fundamentales tienen mayor probabilidad de ocurrir y que un número reducido de causas fundamentales tiene mayor probabilidad de ocurrir en proyectos que implementan ciertos tipos de acciones de conservación. Nuestra taxonomía podría usarse para mejorar el análisis, identificación y aprendizaje subsecuente a partir del fracaso de los esfuerzos de conservación; tratar algunas de las barreras que en la actualidad limitan a los practicantes de la conservación a aprender del fracaso; y contribuir al establecimiento de una cultura efectiva del aprendizaje a partir del fracaso dentro de la conservación.

Endometrial Mesonephric-like Adenocarcinoma Presenting as an Ocular Lesion: A Case Report.

Endometrial mesonephric-like carcinoma (ML-CA) is a recently recognized subtype of aggressive endometrial adenocarcinoma that is morphologically and immunophenotypically similar to mesonephric carcinoma but not typically associated with mesonephric remnants. Here, we report a case of 58-yr-old female who had a past medical history of fibroids and of irregular menstrual bleeding for ~20 yr who presented with visual disturbance. On further investigation, she was found to have a large choroidal peri-papillary tumor of the right eye. A presumptive diagnosis of choroidal melanoma was made. Right eye enucleation was performed, and microscopy revealed moderately differentiated metastatic adenocarcinoma. Further work up was advised. A uterine mass was identified on imaging followed by endometrial biopsy that showed a morphologically and immunohistochemically similar tumor to that in the eye. A hysterectomy was carried out and a malignant neoplasm with varying morphologic patterns including gland formation, solid sheets of tumor cells, cribriform, glomeruloid, spindled and papillary areas was seen. The immunohistochemical profile showed diffuse strong positivity for AE1/AE3, TTF1, P16, and vimentin. CD56, GATA3, Napsin A, and CD10 were focally positive. The neoplastic cells were negative for the following markers ER, PR, WT1, calretinin, and synaptophysin. PDL-1 was negative and mismatch repair protein was proficient. An identical KRAS mutation was detected in both the uterine corpus and ocular tumors. The findings are in keeping with a uterine mesonephric-like adenocarcinoma with an ocular metastasis. An Oncomine Focus-Mutation profile, Thermo-Fisher Scientific Inc., a 60 gene oncologic panel, performed on the ocular tumor, revealed no further mutations.

The relative importance of COVID-19 pandemic impacts on biodiversity conservation globally.

The COVID-19 pandemic has had an enormous impact on almost all aspects of human society and endeavor; the natural world and its conservation have not been spared. Through a process of expert consultation, we identified and categorized, into 19 themes and 70 subthemes, the ways in which biodiversity and its conservation have been or could be affected by the pandemic globally. Nearly 60% of the effects have been broadly negative. Subsequently, we created a compendium of all themes and subthemes, each with explanatory text, and in August 2020 a diverse group of experienced conservationists with expertise from across sectors and geographies assessed each subtheme for its likely impact on biodiversity conservation globally. The 9 subthemes ranked highest all have a negative impact. These were, in rank order, governments sidelining the environment during their economic recovery, reduced wildlife-based tourism income, increased habitat destruction, reduced government funding, increased plastic and other solid waste pollution, weakening of nature-friendly regulations and their enforcement, increased illegal harvest of wild animals, reduced philanthropy, and threats to survival of conservation organizations. In combination, these impacts present a worrying future of increased threats to biodiversity conservation but reduced capacity to counter them. The highest ranking positive impact, at 10, was the beneficial impact of wildlife-trade restrictions. More optimistically, among impacts ranked 11-20, 6 were positive and 4 were negative. We hope our assessment will draw attention to the impacts of the pandemic and, thus, improve the conservation community's ability to respond to such threats in the future.

La pandemia de COVID-19 ha tenido un impacto enorme sobre casi todos los aspectos de la sociedad humana y sus proyectos; el mundo natural y su conservación no han sido la excepción. Por medio de un proceso de consultas a expertos, identificamos y categorizamos en 19 temas y 70 subtemas las maneras en las que la biodiversidad y su conservación han sido o podrían ser afectadas mundialmente por la pandemia. Casi el 60% de los efectos han sido claramente negativos. Posteriormente, creamos un compendio de todos los temas y subtemas, cada uno con textos explicativos, para que en agosto de 2020 un grupo diverso de conservacionistas experimentados con conocimiento de todos los sectores y geografías evaluara cada subtema de acuerdo con su probabilidad de impactar sobre la conservación de la biodiversidad en todo el mundo. Los nueve subtemas con la clasificación más alta tienen un impacto negativo. Estos temas son, en orden de clasificación: los gobiernos dejando de lado al ambiente durante su recuperación económica, reducción de los ingresos basados en el turismo de fauna, incremento en la destrucción de hábitat, financiamiento reducido del gobierno, aumento de la contaminación por plásticos y otros desechos sólidos, debilitamiento de las regulaciones en pro de la naturaleza y su aplicación, incremento en la captura ilegal de animales, disminución de la filantropía y amenazas para la supervivencia de las organizaciones de conservación. La combinación de estos impactos representa un futuro preocupante lleno de amenazas para la conservación de la biodiversidad y una capacidad reducida para contrarrestarlas. El impacto positivo con la clasificación más alta, el 10, fue el impacto benéfico de las restricciones en el mercado de fauna. De manera más optimista, entre los impactos clasificados de los lugares del 11 al 20, seis fueron positivos y cuatro fueron negativos. Esperamos que nuestra evaluación enfoque la atención hacia los impactos de la pandemia y así mejore la habilidad de la comunidad conservacionista para responder a tales amenazas en el futuro. Importancia Relativa de los Impactos de la Pandemia de COVID-19 sobre la Conservación Mundial de la Biodiversidad.

A retrospective comparison of salivary gland fine needle aspiration reporting with the Milan system for reporting salivary gland cytology.

BACKGROUND: Fine needle aspiration (FNA) is a routine sampling method in the diagnostic work up of salivary gland lesions. Despite universal use, no standardised classification existed for the cytopathological reporting of such entities until recently. The Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) aims to standardise the reporting of these lesions, offering risk of malignancy rates and clinical management recommendations. METHODS: We retrospectively applied MSRSGC to cases reported over a 5-year period. Salivary FNA specimens were reclassified according to the MSRSGC as (I) non-diagnostic, (II) non-neoplastic, (III) atypia of undetermined significance (AUS), (IV) benign neoplasm and salivary gland neoplasm of uncertain malignant potential, (V) suspicious for malignancy, and (VI) malignant. Cases with surgical resections were documented and risk of malignancy calculated for each group, where possible. We compared our outcomes with similar studies performed since publication of the Milan criteria. RESULTS: In total, 192 specimens were reassigned as non-diagnostic (n = 30), non-neoplastic (n = 31), AUS (n = 1), benign neoplasm (n = 97) and salivary gland neoplasm of uncertain malignant potential (n = 4), suspicious for malignancy (n = 3), and malignant (n = 26). There were 73 surgical resections. Our calculated risk of malignancy was within the proposed MSRSGC rates for the non-diagnostic, benign neoplasm and malignant groups. One AUS case did not undergo surgery. Benign and malignant sensitivities and specificities for the original reporting categories were 88.24% and 95.72%, and 100% and 95.45% for the MSRSGC, respectively. CONCLUSION: Salivary gland FNA has high diagnostic accuracy and the MSRSGC offers standardised reporting and assistance in the stratification of cases. This may improve communication between pathologists and clinicians with improved outcomes for patients.

Thoracic spinal cord neuromodulation obtunds dorsal root ganglion afferent neuronal transduction of the ischemic ventricle.

Aberrant afferent signaling drives adverse remodeling of the cardiac nervous system in ischemic heart disease. The study objective was to determine whether thoracic spinal dorsal column stimulation (SCS) modulates cardiac afferent sensory transduction of the ischemic ventricle. In anesthetized canines (n = 16), extracellular activity generated by 62 dorsal root ganglia (DRG) soma (T1-T3), with verified myocardial ischemic (MI) sensitivity, were evaluated with and without 20-min preemptive SCS (T1-T3 spinal level; 50 Hz, 90% motor threshold). Transient MI was induced by 1-min coronary artery occlusion (CAO) of the left anterior descending (LAD) or circumflex (LCX) artery, randomized as to sequence. LAD and LCX CAO activated cardiac-related DRG neurons (LAD: 0.15 ± 0.04-1.05 ± 0.20 Hz, P < 0.00002; LCX: 0.08 ± 0.02-1.90 ± 0.45 Hz, P < 0.0003). SCS decreased basal neuronal activity of neurons that responded to LAD (0.15 ± 0.04 to 0.02 ± 0.01 Hz, P < 0.006) and LCX (0.08 ± 0.02 to 0.02 ± 0.01 Hz, P < 0.003). SCS suppressed responsiveness to transient MI (LAD: 1.05 ± 0.20-0.03 ± 0.01 Hz; P < 0.0001; LCX: 1.90 ± 0.45-0.03 ± 0.01 Hz; P < 0.001). Suprathreshold SCS (1 Hz) did not activate DRG neurons antidromically (n = 10 animals). Ventricular fibrillation (VF) was associated with a rapid increase in DRG activity to a maximum of 4.39 ± 1.07 Hz at 20 s after VF induction and a return to 90% of baseline within 10 s thereafter. SCS obtunds the capacity of DRG ventricular neurites to transduce the ischemic myocardium to second-order spinal neurons, a mechanism that would blunt reflex sympathoexcitation to myocardial ischemic stress, thereby contributing to its capacity to cardioprotect.NEW & NOTEWORTHY Aberrant afferent signaling drives adverse remodeling of the cardiac nervous system in ischemic heart disease. This study determined that thoracic spinal column stimulation (SCS) obtunds the capacity of dorsal root ganglia ventricular afferent neurons to transduce the ischemic myocardium to second-order spinal neurons, a mechanism that would blunt reflex sympathoexcitation to myocardial ischemic stress. This modulation does not reflect antidromic actions of SCS but likely reflects efferent-mediated changes at the myocyte-sensory neurite interface.

International evolution of fat, oil and grease (FOG) waste management - A review.

In recent years, issues relating to fat, oil and grease (FOG) in sewer systems have intensified. In the media, sewer blockages caused by FOG waste deposits, commonly referred to as 'fatbergs', are becoming a reminder of the problems that FOG waste can cause when left untreated. These FOG blockages lead to sanitary sewer overflows, property flooding and contamination of water bodies with sewage. Despite these financial and environmentally detrimental effects, a homogenous FOG waste management method has not been developed internationally. However, some successful enduring FOG management programmes have been established, such as in Dublin city and in Scandinavian countries. The aim of this paper is to carry out a review on existing FOG research and management approaches. FOG management involves comprehending: (1) FOG deposition factors in the sewer, (2) FOG prevention and awareness tactics undertaken internationally and (3) potential utilisation methods for FOG waste. This review will highlight that preventing FOG from entering the sewer is the most common approach, often through simple awareness campaigns. The diverted FOG is rarely valorised to bioenergy or biomaterials, despite its potential. Thus, all facets of the FOG waste lifecycle must be identified and managed. Advancements in processes and techniques must be assessed to best determine the future evolution of FOG waste management to assist in achieving a sustainable urban environment.

Heart failure induces changes in acid-sensing ion channels in sensory neurons innervating skeletal muscle.

Heart failure is associated with diminished exercise capacity, which is driven, in part, by alterations in exercise-induced autonomic reflexes triggered by skeletal muscle sensory neurons (afferents). These overactive reflexes may also contribute to the chronic state of sympathetic excitation, which is a major contributor to the morbidity and mortality of heart failure. Acid-sensing ion channels (ASICs) are highly expressed in muscle afferents where they sense metabolic changes associated with ischaemia and exercise, and contribute to the metabolic component of these reflexes. Therefore, we tested if ASICs within muscle afferents are altered in heart failure. We used whole-cell patch clamp to study the electrophysiological properties of acid-evoked currents in isolated, labelled muscle afferent neurons from control and heart failure (induced by myocardial infarction) mice. We found that the percentage of muscle afferents that displayed ASIC-like currents, the current amplitudes, and the pH dose-response relationships were not altered in mice with heart failure. On the other hand, the biophysical properties of ASIC-like currents were significantly different in a subpopulation of cells (40%) from heart failure mice. This population displayed diminished pH sensitivity, altered desensitization kinetics, and very fast recovery from desensitization. These unique properties define these channels within this subpopulation of muscle afferents as being heteromeric channels composed of ASIC2a and -3 subunits. Heart failure induced a shift in the subunit composition of ASICs within muscle afferents, which significantly altered their pH sensing characteristics. These results might, in part, contribute to the changes in exercise-mediated reflexes that are associated with heart failure.

Prognostic significance of tumor budding in rectal cancer biopsies before neoadjuvant therapy.

Tumor budding is an increasingly important prognostic feature for pathologists to recognize. The aim of this study was to correlate intra-tumoral budding in pre-treatment rectal cancer biopsies with pathological response to neoadjuvant chemoradiotherapy and with long-term outcome. Data from a prospectively maintained database were acquired from patients with locally advanced rectal cancer who underwent neoadjuvant chemoradiotherapy. Pre-treatment rectal biopsies were retrospectively reviewed for evidence of intra-tumoral budding. Multivariate logistic regression was used to identify factors contributing to cancer-specific death, expressed as hazard ratios with 95% confidence intervals. Of the 185 patients with locally advanced rectal cancer, 89 patients met the eligibility criteria, of whom 18 (20%) exhibited budding in a pre-treatment tumor biopsy. Intra-tumoral budding predicted a poor pathological response to neoadjuvant chemoradiotherapy (higher ypT stage, P=0.032; lymph node involvement, P=0.018; lymphovascular invasion, P=0.004; and residual poorly differentiated tumors, P=0.005). No patient with intra-tumoral budding exhibited a tumor regression grade 1 or complete pathological response, providing a 100% specificity and positive predictive value for non-response to neoadjuvant chemoradiotherapy. Intra-tumoral budding was associated with a lower disease-free 5-year survival rate (33 vs 78%, P<0.001), cancer-specific 5-year survival rate (61 vs 87%, P=0.021) and predicted cancer-specific death (hazard ratio 3.51, 95% confidence interval 1.03-11.93, P=0.040). Intra-tumoral budding at diagnosis of rectal cancer identifies those who will poorly respond to neoadjuvant chemoradiotherapy and those with a poor prognosis.

The frequencies and clinical implications of mutations in 33 kinase-related genes in locally advanced rectal cancer: a pilot study.

BACKGROUND: Locally advanced rectal cancer (LARC: T3/4 and/or node-positive) is treated with preoperative/neoadjuvant chemoradiotherapy (CRT), but responses are not uniform. The phosphatidylinositol 3-kinase (PI3K), MAP kinase (MAPK), and related pathways are implicated in rectal cancer tumorigenesis. Here, we investigated the association between genetic mutations in these pathways and LARC clinical outcomes. METHODS: We genotyped 234 potentially clinically relevant nonsynonymous mutations in 33 PI3K and MAPK pathway-related genes, including PIK3CA, PIK3R1, AKT, STK11, KRAS, BRAF, MEK, CTNNB1, EGFR, MET, and NRAS, using the Sequenom platform. DNA samples were extracted from pretreatment LARC biopsy samples taken from 201 patients who were then treated with long-course neoadjuvant CRT followed by surgical resection. RESULTS: Sixty-two mutations were detected in 15 genes, with the highest frequencies occurring in KRAS (47 %), PIK3CA (14 %), STK11 (6.5 %), and CTNNB1 (6 %). Mutations were detected in BRAF, NRAS, AKT1, PIK3R1, EGFR, GNAS, MEK1, PDGFRA, ALK, and TNK2, but at frequencies of <5 %. As expected, a pathologic complete response (pCR) was associated with improved 5-year recurrence-free survival (RFS; hazard ratio, 0.074; 95 % CI 0.01-0.54; p = 0.001). Mutations in PI3K pathway-related genes (odds ratio, 5.146; 95 % CI 1.17-22.58; p = 0.030), but not MAPK pathway-related genes (p = 0.911), were associated with absence of pCR after neoadjuvant CRT. In contrast, in patients who did not achieve pCR, mutations in PI3K pathway-related genes were not associated with recurrence-free survival (p = 0.987). However, in these patients, codon 12 (G12D/G12 V/G12S) and 13 mutations in KRAS were associated with poor recurrence-free survival (hazard ratio, 1.579; 95 % confidence ratio, 1.00-2.48; p = 0.048). CONCLUSIONS: Mutations in kinase signaling pathways modulate treatment responsiveness and clinical outcomes in LARC and may constitute rational targets for novel therapies.

Assessing population viability while accounting for demographic and environmental uncertainty.

Predicting the future trend and viability of populations is an essential task in ecology. Because many populations respond to changing environments, uncertainty surrounding environmental responses must be incorporated into population assessments. However, understanding the effects of environmental variation on population dynamics requires information on several important demographic parameters that are often difficult to estimate. Integrated population models facilitate the integration of time series data on population size and all existing demographic information from a species, allowing the estimation of demographic parameters for which limited or no empirical data exist. Although these models are ideal for assessments of population viability, they have so far not included environmental uncertainty. We incorporated environmental variation in an integrated population model to account for both demographic and environmental uncertainty in an assessment of population viability. In addition, we used this model to estimate true juvenile survival, an important demographic parameter for population dynamics that is difficult to estimate empirically. We applied this model to assess the past and future population trend of a rare island endemic songbird, the Montserrat Oriole Icterus oberi, which is threatened by volcanic activity. Montserrat Orioles experienced lower survival in years with volcanic ashfall, causing periodic population declines that were compensated by higher seasonal fecundity in years with high pre-breeding season rainfall. Due to the inclusion of both demographic and environmental uncertainty in the model, the estimated population growth rate in the immediate future was highly imprecise (95% credible interval 0.844-1.105), and the probability of extinction after three generations (in the year 2028) was low (2.1%). This projection demonstrates that accounting for both demographic and environmental sources of uncertainty provides a more realistic assessment of the viability of populations under unknown future environmental conditions.

Application of a Standardized Terminology and Nomenclature for Respiratory Cytology: Experience from a Large Tertiary Respiratory Cancer Centre.

INTRODUCTION: In 2020, the World Health Organization-International Agency for Research on Cancer/International Academy of Cytology (WHO-IARC IAC) joint project was commenced to develop standardized nomenclature and diagnostic criteria in cytopathology internationally. Our institution has been coding all respiratory cytological specimens in a similar fashion for over 10 years. Our aim was to analyse the effectiveness of our respiratory cytology coding system by calculating the estimated risk of malignancy (ROM) and rates of each diagnostic category. METHODS: Over a 2 year period, all endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA), bronchial brushing, bronchial washing, bronchial lavage, and sputum specimens reported at our institution were analysed. For each specimen, the diagnostic code, the relevant indication for each diagnostic procedure, the diagnosis, and the presence or absence of a positive corresponding biopsy were recorded. RESULTS: In total, 1,432 respiratory cytological specimens from 945 patients over a 2-year period were analysed. 467 specimens were confirmed to be associated with a malignant process. The overall ROM for respiratory cytology specimens was 37.7% for nondiagnostic, 18.1% for benign, 46.7% for atypical, 85.7% for suspicious for malignancy, and 91.9% for malignant. For each diagnostic procedure, the ROM increased from the benign to malignant categories. DISCUSSION/CONCLUSION: Our ROM rates for overall respiratory cytology specimens and for EBUS-TBNA, bronchial brushing, and bronchial washing specimens separately are concordant with other major international studies. With the WHO-IARC IAC joint project in progress and an international respiratory cytology coding system being developed, our study has the potential to add value by providing indicative ROM rates, which can be used to inform the development of this new classification system. Our rates of diagnostic accuracy are in keeping with international standards, which support the accuracy of our data.

Neuromodulation targets intrinsic cardiac neurons to attenuate neuronally mediated atrial arrhythmias.

Our objective was to determine whether atrial fibrillation (AF) results from excessive activation of intrinsic cardiac neurons (ICNs) and, if so, whether select subpopulations of neurons therein represent therapeutic targets for suppression of this arrhythmogenic potential. Trains of five electrical stimuli (0.3-1.2 mA, 1 ms) were delivered during the atrial refractory period to mediastinal nerves (MSN) on the superior vena cava to evoke AF. Neuroanatomical studies were performed by injecting the neuronal tracer DiI into MSN sites that induced AF. Functional studies involved recording of neuronal activity in situ from the right atrial ganglionated plexus (RAGP) in response to MSN stimulation (MSNS) prior to and following neuromodulation involving either preemptive spinal cord stimulation (SCS; T(1)-T(3), 50 Hz, 200-ms duration) or ganglionic blockade (hexamethonium, 5 mg/kg). The tetramethylindocarbocyanine perchlorate (DiI) neuronal tracer labeled a subset (13.2%) of RAGP neurons, which also colocalized with cholinergic or adrenergic markers. A subset of DiI-labeled RAGP neurons were noncholinergic/nonadrenergic. MSNS evoked an ∼4-fold increase in RAGP neuronal activity from baseline, which SCS reduced by 43%. Hexamethonium blocked MSNS-evoked increases in neuronal activity. MSNS evoked AF in 78% of right-sided MSN sites, which SCS reduced to 33% and hexamethonium reduced to 7%. MSNS-induced bradycardia was maintained with SCS but was mitigated by hexamethonium. We conclude that MSNS activates subpopulations of intrinsic cardiac neurons, thereby resulting in the formation of atrial arrhythmias leading to atrial fibrillation. Stabilization of ICN local circuit neurons by SCS or the local circuit and autonomic efferent neurons with hexamethonium reduces the arrhythmogenic potential.

Investigative study of hemodilution ratio: 4Vs for vein diameter, valve, velocity, and volumetric blood flow as factors for optimal forearm vein selection for intravenous infusion.

BACKGROUND: Multimodal research and guidelines recognize veins in the forearm used for peripheral intravenous catheter (PIVC) insertion can optimize dwell time. Yet, many PIVCs are still placed in areas of flexion or suboptimal locations such as the back of the hand causing premature failure of >50%. This study identified characteristics of the forearm cephalic vein that make the anatomical location highly successful for PIVC insertion. The goal was to increase the understanding of the human vasculature in association with fluid mechanics in veins above the wrist and below the antecubital fossa. METHODOLOGY: A prospective in-vivo study with 10 consented healthy human volunteers (HHVs) was performed with Color Pulse Wave Doppler Ultrasound that captured high-resolution video and images of vein diameter, velocity of blood flow, and location of venous valves in the forearm. RESULTS: Forearm vein diameter was not directly correlated with higher or lower Velocity of Blood Flow (0.58 cm = 3.0 cm/s). However, Volumetric Blood Flow rates tended to be lower (2.51-8.28 mL/min) with Vein Diameters smaller than 0.29 cm. Ultrasound assessments and Volumetric Blood Flow calculations confirmed natural turbulence in blood and retrograde blood reflux correlated with venous valves opening and closing. Areas of turbulence, with pulse flushing, created backflow with retrograde blood flow around and into the catheter. CONCLUSIONS: Placement of long PIVCs in the cephalic veins of the upper forearm yield adequate flow and hemodilution capacity for veins with at least a 3 to 1 hemodilution ratio. The data from this study, along with previous research, suggest that PIVC placement in the cephalic vein, based on selection criteria, may help to reduce or eliminate intravenous complications such as chemical or mechanical thrombophlebitis causing premature catheter failure. Application of these investigational principles may result in better outcomes and catheter longevity for patients who require intravenous infusions.

The role of botanic gardens in the science and practice of ecological restoration.

Many of the skills and resources associated with botanic gardens and arboreta, including plant taxonomy, horticulture, and seed bank management, are fundamental to ecological restoration efforts, yet few of the world's botanic gardens are involved in the science or practice of restoration. Thus, we examined the potential role of botanic gardens in these emerging fields. We believe a reorientation of certain existing institutional strengths, such as plant-based research and knowledge transfer, would enable many more botanic gardens worldwide to provide effective science-based support to restoration efforts. We recommend botanic gardens widen research to include ecosystems as well as species, increase involvement in practical restoration projects and training practitioners, and serve as information hubs for data archiving and exchange.

Paradoxical Potentiation of Acid-Sensing Ion Channel 3 (ASIC3) by Amiloride via Multiple Mechanisms and Sites Within the Channel.

Acid-Sensing Ion Channels (ASICs) are proton-gated sodium-selective cation channels that have emerged as metabolic and pain sensors in peripheral sensory neurons and contribute to neurotransmission in the CNS. These channels and their related degenerin/epithelial sodium channel (DEG/ENaC) family are often characterized by their sensitivity to amiloride inhibition. However, amiloride can also cause paradoxical potentiation of ASIC currents under certain conditions. Here we characterized and investigated the determinants of paradoxical potentiation by amiloride on ASIC3 channels. While inhibiting currents induced by acidic pH, amiloride potentiated sustained currents at neutral pH activation. These effects were accompanied by alterations in gating properties including (1) an alkaline shift of pH-dependent activation, (2) inhibition of pH-dependent steady-state desensitization (SSD), (3) prolongation of desensitization kinetics, and (4) speeding of recovery from desensitization. Interestingly, extracellular Ca2+ was required for paradoxical potentiation and it diminishes the amiloride-induced inhibition of SSD. Site-directed mutagenesis within the extracellular non-proton ligand-sensing domain (E79A, E423A) demonstrated that these residues were critical in mediating the amiloride-induced inhibition of SSD. However, disruption of the purported amiloride binding site (G445C) within the channel pore blunted both the inhibition and potentiation of amiloride. Together, our results suggest that the myriad of modulatory and blocking effects of amiloride are the result of a complex competitive interaction between amiloride, Ca2+, and protons at probably more than one site in the channel.

An update of the Worldwide Integrated Assessment (WIA) on systemic insecticides. Part 2: impacts on organisms and ecosystems.

New information on the lethal and sublethal effects of neonicotinoids and fipronil on organisms is presented in this review, complementing the previous Worldwide Integrated Assessment (WIA) in 2015. The high toxicity of these systemic insecticides to invertebrates has been confirmed and expanded to include more species and compounds. Most of the recent research has focused on bees and the sublethal and ecological impacts these insecticides have on pollinators. Toxic effects on other invertebrate taxa also covered predatory and parasitoid natural enemies and aquatic arthropods. Little new information has been gathered on soil organisms. The impact on marine and coastal ecosystems is still largely uncharted. The chronic lethality of neonicotinoids to insects and crustaceans, and the strengthened evidence that these chemicals also impair the immune system and reproduction, highlights the dangers of this particular insecticidal class (neonicotinoids and fipronil), with the potential to greatly decrease populations of arthropods in both terrestrial and aquatic environments. Sublethal effects on fish, reptiles, frogs, birds, and mammals are also reported, showing a better understanding of the mechanisms of toxicity of these insecticides in vertebrates and their deleterious impacts on growth, reproduction, and neurobehaviour of most of the species tested. This review concludes with a summary of impacts on the ecosystem services and functioning, particularly on pollination, soil biota, and aquatic invertebrate communities, thus reinforcing the previous WIA conclusions (van der Sluijs et al. 2015).

Improving tumor budding reporting in colorectal cancer: a Delphi consensus study.

Tumor budding is a long-established independent adverse prognostic marker in colorectal cancer, yet methods for its assessment have varied widely. In an effort to standardize its reporting, a group of experts met in Bern, Switzerland, in 2016 to reach consensus on a single, international, evidence-based method for tumor budding assessment and reporting (International Tumor Budding Consensus Conference [ITBCC]). Tumor budding assessment using the ITBCC criteria has been validated in large cohorts of cancer patients and incorporated into several international colorectal cancer pathology and clinical guidelines. With the wider reporting of tumor budding, new issues have emerged that require further clarification. To better inform researchers and health-care professionals on these issues, an international group of experts in gastrointestinal pathology participated in a modified Delphi process to generate consensus and highlight areas requiring further research. This effort serves to re-affirm the importance of tumor budding in colorectal cancer and support its continued use in routine clinical practice.