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Gene expression analysis is critical to precisely characterize complex tissues and provide insight into a disease condition. Techniques like PCR, sequencing, and northern blotting are highly sensitive and specific but are unable to provide information about spatial positioning of target genes. Visualization of gene expression with a spatial context can be critical in identifying complex milieus in heterogenous tissues like tumors. The RNAscope in situ hybridization (ISH) technology detects target RNA expression with high sensitivity and specificity at single-cell resolution. To understand the cellular cross talk between different cell populations, it is essential to simultaneously study gene and protein expression within a complex tissue. This chapter details combining the RNAscope ISH assay with immunofluorescence (IF) in one protocol to simultaneously visualize gene expression and protein expression in human tumor tissue and mouse brain tissue.

This paper addresses the problem of safe navigation in an environment with randomly placed static obstacles. We convert a commercial powered wheelchair into a semiautonomous vehicle with limited sight (environment awareness), by instrumenting it using off-the-shelf ultrasonic sensors and associated electronic boards (Arduino). In the continuity of our previous work where we had used stochastic dynamic programming to formulate optimization problems which led to relatively large size look-up tables that can be used as supervisory control, here we propose to extract rules using those results (that data). The advantage of this approach is a lowcomputational cost for future online implementation, and the drawback is a suboptimal policy. The feasibility is assessed by running simulations in a fairly realistic environment (Unity3D).

International audience; OBJECTIVE:Insulo-opercular seizures are characterized by diverse semiology, related to the insula's multiple functional roles and extensive connectivity. We aimed to identify semiologic subgroups and correlate these with insulo-opercular subregions based on connectional architecture.METHODS:We retrospectively collected a large series of 37 patients with insulo-opercular seizures explored by stereoelectroencephalography (SEEG) from three epilepsy centers. A new human brain atlas (Brainnetome Atlas, BNA) based on both anatomic and functional connections was employed to segment insulo-opercular cortex. Semiology and SEEG changes were carefully reviewed and quantified. Principal component analysis and cluster analysis were used to correlate semiologic characteristics with insulo-opercular subregions.RESULTS:Four main semiologic subgroups were identified, organized along an anteroventral to posterodorsal axis based on BNA. Group 1 was characterized by epigastric sensation and/or integrated gestural motor behaviors with or without feelings of fear or rage, involving the anteroventral insular regions and mesial temporal lobes. Group 2 was characterized by auditory sensations and symmetric proximal/axial tonic signs involving the posteroventral temporal operculum. The characteristics of group 3 were orofacial and laryngeal signs, involving the intermediate insulo-opercular regions. The features of group 4 were somatosensory signs followed by nonintegrated gestural motor behaviors and/or asymmetric tonic signs involving the posterodorsal insulo-opercular regions with propagation to the mesial frontal lobes. Thus anteroventral seizure organizations predominantly showed limbic system semiology, whereas more posterodorsal regions were associated with semiology involving mainly the sensorimotor system. Subjective symptoms proved to be particularly discriminating factors.SIGNIFICANCE:Insulo-opercular seizures can be categorized in terms of clinical semiology and correlate with connectional architecture subregions along an anteroventral-posterodorsal axis in line with the cytoarchitectonic gradient rather than the gyral anatomy of the insula cortex. This provides new insights into facilitating differential diagnosis and presurgical localization but also highlights the importance of considering connectional architecture in determining neural correlates of complex semiologic patterns.

Early infantile epileptic encephalopathy type 28 is a refractory epilepsy with early onset, poor prognosis, and hereditary causes. WW domain-containing oxidoreductase (WWOX) gene mutation can result in epileptic encephalopathy, but the mechanism remains unclear. We present the case of a patient with epilepsy and WWOX compound heterozygous mutations. The seizures manifested as tonic-clonic, convulsive and were refractory to drugs. Magnetic resonance imaging showed a widened subarachnoid space and thin corpus callosum. The patient died from asphyxia at the age of one year and 23 days. Peripheral blood was taken from the patient and his parents, and whole-exome sequencing was investigated to determine possible gene mutation. Two compound heterozygous mutations were identified: c.172+1GC (with no amino acid change) and c.984CG (amino acid change: p.Tyr328Ter). The pathophysiology of epileptic encephalopathy related to the WWOX gene remains to be determined, and further studies are required to elucidate possible mechanisms.

Gene expression analysis is critical to precisely characterize complex tissues and provide insight into a disease condition. Techniques like PCR, sequencing, and northern blotting are highly sensitive and specific but are unable to provide information about spatial positioning of target genes. Visualization of gene expression with a spatial context can be critical in identifying complex milieus in heterogenous tissues like tumors. The RNAscope in situ hybridization (ISH) technology detects target RNA expression with high sensitivity and specificity at single-cell resolution. To understand the cellular cross talk between different cell populations, it is essential to simultaneously study gene and protein expression within a complex tissue. This chapter details combining the RNAscope ISH assay with immunofluorescence (IF) in one protocol to simultaneously visualize gene expression and protein expression in human tumor tissue and mouse brain tissue.

BACKGROUND Ventilation is critical in airway management, and failure can be fatal. The optimal ventilation approach for endotracheal intubation in children with difficult airways remains controversial. The Pediatric Difficult Intubation (PeDI) Registry is an international multicenter registry that collects intubation data in difficult to intubate children. The registry captures the initial (at induction) and final ventilation technique (at intubation), the use of neuromuscular blocking drugs (NMBDs), airway reactivity during intubation, and complications. We analyzed data in the PeDI Registry to determine the frequency of use of various ventilation techniques and associated complications. Because spontaneously breathing patients ventilate throughout intubation, we hypothesized that spontaneous ventilation would be associated with fewer complications than other approaches. METHODS We queried the PeDI Registry for cases entered between September 2012 and February 2016, from 16 children's hospitals. We categorized the attending anesthesiologist's ventilation plan into 3 groups: spontaneous ventilation, controlled ventilation after administering an NMBD, and controlled ventilation without administering an NMBD. Generalized Estimating Equation (GEE) model, with a binomial family distribution and logit link, was used to determine the association between ventilation technique and the risk of complications, as well as to account for within-site clustering. Propensity score matching was further applied to balance pretreatment characteristics of ventilation groups. RESULTS Of 1289 anticipated difficult intubations, 507 (39%) were managed with spontaneous ventilation, 453 (35%) controlled ventilation with an NMBD, and 329 (26%) controlled ventilation without an NMBD. Complications occurred in 242 (18.8%; 95% confidence interval [CI], 16.6%-20.9%) patients. Of these, 218 (16.9%) were nonsevere, and 24 (1.9%) were severe. The spontaneous ventilation group had 114 (22.5%, standardized residual [Std.Res] = 4.29) nonsevere complications, which was higher than the controlled ventilation with an NMBD 60 (13.3%, Std.Res = -2.58), and controlled ventilation without an NMBD 44 (13.4%, Std.Res = -1.98), P < .001. Nearest neighbor matching with caliper width equal to 0.2 of the standard deviation (SD) of the logit of the propensity score also demonstrated that patients with spontaneous ventilation had greater odds of complications compared to controlled ventilation techniques: odds ratio (OR) = 2.07 (95% CI, 1.36-3.15; P = .001). CONCLUSIONS Spontaneous ventilation is associated with more nonsevere complications, such as hypoxemia and laryngospasm, than controlled ventilation techniques during intubation of children with difficult airways. Inadequate anesthetic depth may contribute to increased complications.

This article presents progress made towards implementing a shared control framework for a smart wheelchair based upon stochastic dynamic programming (a model-based control design). First, we describe the mechanical, electrical and software design process of our instrumented wheelchair platform. Then, we detail a deterministic control-oriented model of the wheelchair motion dynamics using Euler-Lagrange equations. Finally, we discuss the development of a stochastic model of the human driver’s intention in view of using Markov chain.

Rhodium was reported to be an essential component of the efficient cocatalyst (Rh2−xCrxO3 or core/shell of Rh@Cr2O3) on GaN–ZnO for photocatalytic overall water splitting (POWS). Herein we demonstrate that the Rh can be replaced by less-expensive Pd deposited via atomic layer deposition and together with photodeposition of Cr2O3 shell, a comparable activity in POWS can be achieved even with much less Pd loading than Rh in Rh2−xCrxO3/GaN–ZnO under the same reaction conditions.

BackgroundThe tumor microenvironment (TME) is highly complex, comprised of tumor cells, immune cells, stromal cells, and extracellular matrix. Understanding spatial interactions between various cell types and their activation states in the TME is crucial for implementing successful immunotherapy strategies against various types of cancer. This study demonstrates a highly sensitive and specific multiplexed technique, the RNAscope HiPlex v2 in situ hybridization (ISH) assay for spatial and transcriptomic profiling of target genes to assess immune regulation in human lung, breast, cervical and ovarian FFPE tumor tissues.MethodsWe have expanded our current RNAscope HiPlex assay capability of iteratively multiplexing up to 12 targets in fixed and fresh frozen samples to include formalin fixed paraffin embedded (FFPE) tissues. The novel FFPE reagent effectively reduces background autofluorescence, improving the signal to noise ratio. We have leveraged this technology to investigate spatial expression of 12 oncology and immuno-oncology target genes, including tumor markers, immune checkpoint markers, immunosuppression markers, immune cell markers and secreted chemokine RNA expression profile within the TME. The targets were simultaneously registered using HiPlex image registration software v2 that enables background subtraction.ResultsWe visualized T cell infiltration and identified T cell subsets within tumors using CD3and CD8 expression and activated T cells by IFNG expression. We further identified subsets of pro- and anti-inflammatory macrophages by CD68 and CD163 expression as well effector cells which secrete chemokines and cytokine. We also detected the hypoxia markers HIF1A and VEGF to elucidate the immunosuppressive state of tumor cells. Preliminary analysis and quantification of the HIF1A expression using HALO® image analysis software showed higher copy numbers in the lung tumor as compared to the other tumors, demonstrating the sensitivity of the assay through differential expression. We additionally showed the differential expression of immune checkpoint markers PDCD1, and CD274 within the TME.ConclusionsUsing a highly sensitive multiplexed RNAscope HiPlex v2 ISH assay, we have demonstrated the capability of this technique to spatially resolve 12 targets in four different tumor types. The FFPE reagent efficiently quenched background autofluorescence in the tissues and identified immune cell signatures within the TME. Quantification of immunosuppressive markers further depicted a differential expression among various tumors. This technology is highly beneficial for investigating complex and spatial tumor-stroma interactions in basic science and translational research. The assay can also provide valuable understanding of the biological crosstalk among various cell types in complex and heterogeneous tissues.

Genomic DNA anomalies such as copy number variations (gene duplication, amplification, deletion) and gene rearrangements are important biomarkers and drug targets in many cancer types. DNA in-situ hybridization (ISH) is the gold standard method to directly visualize these molecular alterations in formalin-fixed paraffin-embedded (FFPE) tumor tissues at single-cell resolution within a histological section. However, currently available fluorescent ISH (FISH) assays provide limited morphological detail due to the use of fluorescent nuclear staining compared to chromogenic staining. Furthermore, FISH techniques rely on expensive fluorescence microscopes, risk loss of fluorescent signal over time and involve tedious imaging at high magnifications (100X). There is thus an unmet need for a sensitive and robust chromogenic DNA-ISH assay that can enable high-resolution detection of genomic DNA targets with the ease of bright-field microscopy. We present here DNAscope - a novel chromogenic DNA-ISH assay - for detecting and visualizing genomic DNA targets under a standard light microscope. DNAscope is based on the widely used RNAscope® double-Z probe design and signal amplification technology and provides unparalleled sensitivity and specificity with large signal dots readily visualized at 40X magnification and with full morphological context. Furthermore, DNAscope ensures specific DNA detection without interference from RNA due to the use of a novel RNA removal method. Using a duplex chromogenic detection assay in red and blue, we demonstrate highly specific and efficient detection of gene rearrangements (ALK, ROS1, RET and NTRK1), gene amplification (ERBB2, EGFR, MET) and deletion (TP53 and CDKN2A). The DNAscope assay has been carefully optimized for probe signal size and color contrast to enable easy interpretation of signal patterns under conventional light microscopy or digital pathology. Compared to conventional FISH assays, DNAscope probes are standard oligos that are designed in silico to be free of any repetitive sequences and can be rapidly synthesized for any DNA target. In conclusion, the DNAscope assay provides a powerful and convenient alternative to commonly used FISH assays in many cancer research applications. Citation Format: Li-Chong Wang, Farzaneh Tondnevis, Courtney Todorov, Jayson Gaspar, Aparna Sahajan, Vasudha Murlidhar, Bingqing Zhang, Xiao-Jun Ma. DNAscopeTM: A novel chromogenic in-situ hybridization technology for high-resolution detection of DNA copy number and structural variations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB190.

Complex tissues such as tumors are comprised of multiple cells types and extracellular matrix. These cells include heterogenous populations of immune cells that infiltrate the tumors. Understanding the composition of these immune infiltrates in the tumor microenvironment (TME) can provide key insights to guide therapeutic intervention and predict treatment response. Thorough understanding of complex tissue dynamics and immune cell characterization requires a multi-omics approach. Simultaneous detection of RNA and protein using in situ hybridization (ISH) and immunohistochemistry/immunofluorescence (IHC/IF) can reveal cellular sources of secreted proteins, identify specific cell types, and visualize the spatial organization of cells within the tissue. However, a sequential workflow of ISH followed by IHC/IF frequently yields suboptimal protein detection because the protease digestion step in the ISH protocol resulting in poor antibody signal. Here we demonstrate a newly developed integrated ISH/IHC workflow that can substantially improve RNA-protein co-detection, enabling the visualization and characterization of tumor immune infiltrates at single-cell resolution with spatial and morphological context. To characterize tumor-infiltrating immune cells in a tumor TMA (tumor microarray), we utilized the RNAscope Multiplex Fluorescence assay in combination with the RNA-Protein Co-detection Kit to detect multiple immune cell populations. Immune cells such as macrophages, T cells and NK cells were detected using specific antibodies against CD68, CD8, CD4 and CD56, respectively. Precise characterization of these immune cells was achieved by using probes against targets such as CCL5, IFNG, GNZB, IL-12, NCR1 etc. that not only help in identifying specific immune cells but also assist in determining their activation states. We identified subsets of T cells such as CD4+ regulatory T cells and CD8+ cytotoxic T lymphocytes. Additionally, we were able to determine the activation states of CD8+ T cells by visualizing the expression of IFNG and GZMB. Furthermore, infiltrating macrophages were identified by detecting the CD68 protein expression while the M1 and M2 subsets were differentiated by detecting the M2-specific target RNA for CD163. Similarly, NK cells were identified by detecting CD56 protein in combination with CCL5 and NCR1 RNA expression. Interestingly, the degree of infiltration of the different immune cell populations varied based on the tumor type. In conclusion, the new RNAscope-ISH-IHC co-detection workflow and reagents enable optimized simultaneous visualization of RNA and protein targets by enhancing the compatibility of antibodies - including many previously incompatible antibodies - with RNAscope. This new workflow provides a powerful new approach to identifying and characterizing tumor infiltrating populations of immune cells. Citation Format: Anushka Dikshit, Jyoti Phatak, Lydia Hernandez, Emerald Doolittle, Vasudha Murlidhar, Bingqing Zhang, Xiao-Jun Ma. Characterizing tumor-infiltrated immune cells with spatial context using an integrated RNAscope-immunohistochemistry co-detection workflow in FFPE tissues [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB235.

Conventionally, it is common that video retrieval methods aggregate the visual feature representations from every frame as the feature of the video, where each frame is treated as an isolated, static image. Such methods lack the power of modeling the intra-frame and inter-frame relationships for the local regions, and are often vulnerable to the visual redundancy and noise caused by various types of video transformation and editing, such as adding image patches, adding banner, etc. From the perspective of video retrieval, a video’s key information is more often than not convoyed by geometrically centered, dynamic visual content, and static areas often reside in regions that are farther from the center and often exhibit heavy visual redundancies temporally. This phenomenon is hardly investigated by conventional retrieval methods. In this article, we propose an unsupervised video retrieval method that simultaneously models intra-frame and inter-frame contextual information for video representation with a graph topology that is constructed on top of pyramid regional feature maps. By decomposing a frame into a pyramid regional sub-graph, and transforming a video into a regional graph, we use graph convolutional networks to extract features that incorporate information from multiple types of context. Our method is unsupervised and only uses the frame features extracted by pre-trained network. We have conducted extensive experiments and have demonstrated that the proposed method outperforms state-of-the-art video retrieval methods.

Characterizing heterogenous populations of tumor-infiltrating immune cells requires a multi-omics approach. Here we demonstrate a newly developed integrated in situ hybridization (ISH) and immunohistochemistry (IHC/IF) workflow that can substantially improve RNA-protein co-detection, enabling the visualization and characterization of tumor immune infiltrates at single-cell and spatial resolution. To characterize tumor-infiltrating immune cells in a tumor TMA (tumor microarray), we utilized the RNAscope Multiplex Fluorescence assay in combination with the RNA-Protein Co-detection Kit to detect multiple immune cell populations. Immune cells such as macrophages, T cells and NK cells were detected using antibodies against CD68, CD8, CD4 and CD56 in combination with probes targeting CCL5, IFNG, GNZB, IL-12, NCR1 etc. We identified CD4+ regulatory T cells and CD8+ cytotoxic T lymphocytes. Additionally, we determine the activation states of CD8+ T cells by visualizing IFNG and GZMB expression. Furthermore, infiltrating macrophages were detected by CD68 protein expression while the M1 and M2 subsets were differentiated by using the M2-specific marker, CD163. NK cells were identified by detecting CD56 protein in combination with CCL5 and NCR1 RNA expression. The degree of immune cell infiltration varied based on the tumor type. In conclusion, the new RNAscope-ISH-IHC co-detection workflow and reagents enable optimized simultaneous visualization of RNA and protein targets by enhancing the compatibility of antibodies, including many previously incompatible antibodies with RNAscope . This new workflow provides a powerful approach to identifying and characterizing tumor infiltrating immune cells.

Acute heart failure due to myocarditis is not common in Takayasu arteritis, let alone in combination with thrombosis affecting both ventricles and pulmonary arteries. The concomitant infection of non-tuberculosis mycobacterium further complicates the clinical scenario and poses challenges for implementation of tailored treatments. This case report describes a teenage girl with a history of intermittent claudication and Erythema Nodosum who developed acute heart failure. Detailed clinical investigations and imaging techniques confirmed the diagnosis.

A three-dimensional, low platinum (Pt) catalyst was prepared by decorating platinum on the palladium nanoflowers (Pd NF) by an underpotential deposition (UPD) method. The Pd NF was synthesized by a solvothermal approach, using oleic acid as the template and benzyl alcohol as the solvent-reducing agent. The obtained Pd with a morphology of uniform nanoflowers is composed of plentiful nanosheets. After decorating with platinum, the catalyst Pd NF@Pt exhibits much higher activity for the oxygen reduction reaction (ORR) compared to commercial Pt/C (Pt 20 wt%). The interaction between deposited Pt and Pd NF was revealed by XPS analysis, and the high performance of the Pd NF@Pt catalyst was attributed to following two aspects: the increased of dispersion of platinum based on Pd NF substrate, and the increased intrinsic activity of the active sites caused by the interaction of Pt and Pd NF.

The oxygen reduction reaction (ORR) is a key step in H2–O2 fuel cells, which, however, suffers from slow kinetics even for state-of-the-art catalysts. In this work, by making use of photocatalysis, the ORR was significantly accelerated with a polymer semiconductor (polyterthiophene). The onset potential underwent a positive shift from 0.66 to 1.34 V, and the current was enhanced by a factor of 44 at 0.6 V. The improvement was further confirmed in a proof-of-concept light-driven H2–O2 fuel cell, in which the open circuit voltage (Voc) increased from 0.64 to 1.18 V, and the short circuit current (Jsc) was doubled. This novel tandem structure combining a polymer solar cell and a fuel cell enables the simultaneous utilization of photo- and electrochemical energy, showing promising potential for applications in energy conversion and storage.

ObjectivesMillions of children die every year from serious childhood illnesses. Most deaths are avertable with access to quality care. Saving Children’s Lives (SCL) includes an abbreviated high-intensity training (SCL-aHIT) for providers who treat serious childhood illnesses. The objective of this study was to examine the impact of SCL-aHIT on knowledge acquisition and retention of providers.Setting76 participating centres who provide primary and secondary care in Kweneng District, Botswana.ParticipantsDoctors and nurses expected by the District Health Management Team to provide initial care to seriously ill children, completed SCL-aHIT between January 2014 and December 2016, submitted demographic data, course characteristics and at least one knowledge assessment.MethodsRetrospective, cohort study. Planned and actual primary outcome was adjusted acquisition (change in total knowledge score immediately after training) and retention (change in score at 1, 3 and 6 months), secondary outcomes were pneumonia and dehydration subscores. Descriptive statistics and linear mixed models with random intercept and slope were conducted. Relevant institutional review boards approved this study.Results211 providers had data for analysis. Cohort was 91% nurses, 61% clinic/health postbased and 45% pretrained in Integrated Management of Childhood Illness (IMCI). A strong effect of SCL-aHIT was seen with knowledge acquisition (+24.56±1.94, pConclusionsaHIT for care of the seriously ill child significantly increased provider knowledge and loss of knowledge occurred over time. IMCI training did not significantly impact overall knowledge acquisition nor retention, while professional status impacted overall score and lost to follow-up impacted retention.

Deviations (DEVs) from resuscitation guidelines are associated with worse outcomes after adult in-hospital cardiac arrest (IHCA), but impact during pediatric IHCA is unknown.Retrospective cohort study of prospectively collected data from the American Heart Association's Get With The Guidelines-Resuscitation registry. Children who had an index IHCA of ≥1 min from 2000 to 2014 were included. DEVs are defined by the registry by category (airway, medications, etc.) A composite measure termed circulation DEV(C-DEV), defined as at least one process deviation in the following categories: medications, defibrillation, vascular access, or chest compressions, was the primary exposure variable. Primary outcome was survival to hospital discharge. Mixed-effect models with random intercept for each hospital assessed the relationship of DEVs with survival to hospital discharge. Robustness of findings was assessed via planned secondary analysis using propensity score matching.Among 7078 eligible index IHCA events, 1200 (17.0%) had DEVs reported. Airway DEVs (466; 38.8%) and medication DEVs (321; 26.8%) were most common. C-DEVs were present in 629 (52.4%). Before matching, C-DEVs were associated with decreased rate of ROSC (aOR = 0.53, CI95: 0.43-0.64, p 0.001) and survival to hospital discharge (aOR = 0.71, CI95: 0.60-0.86, p 0.001). In the matched cohort (C-DEV n = 573, no C-DEV n = 1146), C-DEVs were associated with decreased rate of ROSC (aOR 0.76, CI95 0.60-0.96, p = 0.02), but no association with survival to hospital discharge (aOR 1.01, CI95 0.81-1.25, p = 0.96).DEVs were common in this cohort of pediatric IHCA. In a propensity matched cohort, while survival to hospital discharge was similar between groups, events with C-DEVs were less likely to achieve ROSC.

To meet the requirements of national “Double first-class” construction plan, business schools strive to focus resources on establishing characteristic and preponderant disciplines as the “Double first-class” development goal during the 13th Five-Year Plan or even longer periods. In the trend of globalization, improving discipline construction in virtue of international accreditation in business education has become an important tool for business schools to improve education quality and enhance domestic and international reputation. Considering the effects of accreditation effort on school reputation and student skill improvement, this paper mainly studies accreditation mode selection for domestic business schools in the process of business education accreditation. The results show that: 1) accreditation effort across different accreditation modes shows performance of significant differences in improving school reputation; and 2) business education accreditation has positive impact on both school reputation and student skill improvement, thus uplifts the competitiveness of business school. This paper appears to be the first to study business education accreditation of business schools with quantitative analysis, and provides significant theoretical guidance for domestic business schools to promote the construction of first-class disciplines.

Obstructive sleep apnea (OSA) has a prevalence of 4% in children. Few studies have explored the role of secondhand smoke (SHS) on OSA severity and have shown contradicting results. Most studies have focused on the effect of SHS on snoring. This study explored the association of SHS exposure and OSA severity in children aged 3-18 years.This is a retrospective single center IRB-approved study. Electronic Medical Records (EMR) were queried between 1/24/2015 and 1/24/2018 to obtain data on SHS exposure with standard questionnaires from perioperative database. SHS was analyzed as a binary variable and OSA was measured using obstructive apnea hypopnea index (OAHI) from polysomnography (PSG) as a continuous variable. Analyses were done on all children and in those with severe OSA (OAHI≥10/h) as a subgroup.EMR query yielded 101,884 children of whom 3776 had PSG. Limiting baseline PSG in 3-18-year-old and reliable information on SHS yielded 167 analyzable children of whom 70 had severe OSA. Children exposed to SHS had significantly more public insurance than non-exposed (p 0.0001). Among children with severe OSA, median OAHI was significantly higher in SHS exposed compared to non-exposed (29.0vs.19.5,p = 0.04), but not across all children. In multivariable analysis SHS exposure increased OAHI by 48% in severe OSA subgroup (95%CI: 8%-102%; p = 0.01) when adjusted for race, body mass index, and adjusted household income.Children aged 3-18 years with severe OSA who were exposed to SHS were found to have 1.48 increase in odds of OAHI than those without SHS exposure. Results could be limited by retrospective nature of study and EMR tools.

BACKGROUND In infants and young children, anesthetic dosing is based on population pharmacokinetics and patient hemodynamics not on patient-specific brain activity. Electroencephalography (EEG) provides insight into brain activity during anesthesia. The primary goal of this prospective observational pilot study was to assess the prevalence of isoelectric EEG events-a sign of deep anesthesia-in infants and young children undergoing general anesthesia using sevoflurane or propofol infusion for maintenance. METHODS Children 0-37 months of age requiring general anesthesia for surgery excluding cardiac, intracranial, and emergency cases were enrolled by age: 0-3, 4-6, 7-12, 13-18, and 19-37 months. Anesthesia was maintained with sevoflurane or propofol infusion. EEG was recorded from induction to extubation. Isoelectric EEG events (amplitude

The purpose of this paper is to observe the protective action and its effective mechanism of eriodictyol on lipopolysaccharide (LPS)-induced acute lung injury (ALI). In this study, our results indicated that eriodictyol could dramatically suppress the inflammatory mediators, including interleukin-6 (IL-6), IL-1β, prostaglandin E2, and tumor necrosis factor-α in bronchoalveolar lavage fluid of LPS-challenged mice. Eriodictyol also alleviated the wet/dry ratio and improved pathological changes of the lung. In addition, eriodictyol significantly decreased myeloperoxidase activity and malondialdehyde content as well as increased superoxide dismutase activity. Moreover, eriodictyol inhibited the COX-2/NLRP3/NF-κB signaling pathway in the lung tissues of ALI mice. In conclusion, our observations validated that eriodictyol processed the protective effects on ALI mice, which was related to the regulation of the COX-2/NLRP3/NF-κB signaling pathway.

AIM To compare the diagnostic performance of multiparametric MRI (mpMRI) parameters for Gleason score (GS) and cellularity metrics of prostate cancer (PCa) in the peripheral zone (PZ) and transition zone (TZ) separately. MATERIALS AND METHODS In total, 225 PCa patients with preoperative mpMRI and whole-mount pathological sections were enrolled retrospectively. Detection rates of index lesions (highest GS or largest dimension) and clinically significant PCa (csPCa) were evaluated. Tumour-to-muscle ratio and skewness of T2 signal intensity, average apparent diffusion coefficient (ADCmean) and 10th percentile ADC (ADC10%) were derived and correlation with GS was performed with Spearman's correlation coefficient (ρ), while effectiveness in differentiating GS 6 from GS ≥7 was compared with receiver operating characteristic (ROC) analysis. Moreover, correlation of cellularity metrics with mpMRI parameters was evaluated with Pearson's correlation coefficient (r). RESULTS In total, 398 lesions were identified, with 87.1% (196/225) index lesions and 86.8% (249/287) csPCa detected. Compared to T2 parameters, ADC parameters, especially ADCmean, correlated better with GS (maximal ρ: –0.58 versus –0.33, p=0.011) and yielded significantly higher area under the curve (AUC) in differentiating GS 6 from GS ≥7 (maximal AUC: 0.854 versus 0.731, p=0.020) among PZ lesions. Moreover, ADCmean demonstrated significantly moderate correlation with the nuclear-to-cytoplasmic ratio and nuclear fraction (r=–0.403 and –0.514, p CONCLUSION mpMRI could effectively detect index and csPCa lesions. ADC parameters, especially ADCmean, correlated better with GS and cellularity metrics than T2 in PZ, while all parameters demonstrated poor performance within TZ lesions.

OBJECTIVES Estimate the inter-rater reliability of critical care nurses performing a pediatric modification of the Glasgow Coma Scale in a contemporary PICU. DESIGN Prospective observation study. SETTING Large academic PICU. PATIENTS/SUBJECTS All 274 nurses with permanent assignments in the PICU were eligible to participate. A subset of 18 nurses were selected as study registered nurses. All PICU patients were eligible to participate. INTERVENTIONS None. MEASUREMENTS AND MAIN RESULTS PICU nurses were educated and demonstrated proficiency on a pediatric modification of the Glasgow Coma Scale we created to make it more applicable to a diverse PICU population that included patients who are sedated, mechanically ventilated, and/or have developmental disabilities. Each study registered nurse observed a sample of nurses perform the Glasgow Coma Scale, and they independently scored the Glasgow Coma Scale. Patients were categorized as having developmental disabilities if their preillness Pediatric Cerebral Performance Category score was greater than or equal to 3. Fleiss' Kappa (κ), intraclass correlation coefficient, and percent agreement assessed inter-rater reliability for each Glasgow Coma Scale component (eye, verbal, motor) and age-specific scale (≥ 2 and < 2-yr-old). The overall percent agreement between study registered nurses and nurses was 89% for the eye, 91% for the verbal, and 79% for the motor responses. Inter-rater reliability ranged from good (intraclass correlation coefficient = 0.75) to excellent (intraclass correlation coefficient = 0.96) for testable patients. Agreement on the motor response was significantly lower for children with developmental disabilities (< 2 yr: 59% vs 95%; p = 0.0012 and ≥ 2 yr: 55% vs 91%; p = 0.0012). Agreement was significantly worse for intermediate range Glasgow Coma Scale motor responses compared with responses at the extremes (e.g., motor responses 2, 3, 4 vs 1, 5, 6; p < 0.05). CONCLUSIONS A pediatric modification of the Glasgow Coma Scale performed by trained PICU nurses has excellent inter-rater reliability, although reliability was reduced in patients with developmental disabilities and for intermediate range Glasgow Coma Scale responses. Further research is needed to determine the effectiveness of this Glasgow Coma Scale modification to detect clinical deterioration.

ObjectiveTo determine trends in opioid prescribing for home use after pediatric outpatient surgery.DesignRetrospective analysis of a de-identified database.SettingMultispecialty children’s hospital and freestanding surgery centers.Patients, ParticipantsA total of 65,190 encounters of pediatric outpatient surgeries from 2013 through 2017 for nine different surgical specialties. Patients in the cardiothoracic service and nonpainful procedures were excluded.MainOutcome Measures. Incidence rate of prescribing, dose, number of doses available (i.e., duration of therapy), and maximum weight-based home opioid availability from 2013 to 2017. Additional independent variables included sex, age, weight, race/ethnicity, insurance type (private vs public), and surgical service.ResultsThe incidence rate of receiving a take-home opioid prescription at discharge ranged from 18% to 21% between 2013 and 2017, with no clear directional trend. Among patients prescribed opioids, however, the maximum available take-home dose steadily declined from 2013 through 2017 (P ConclusionsThe rate of receiving a take-home opioid prescription and the dose prescribed remained stable from 2013 to 2017, but the duration of treatment steadily declined, and beginning in 2015, the amount per dose also decreased. Certain subgroups of patients were more likely to be prescribed opioids and will require further investigation and confirmation.

The photoelectrochemical (PEC) method offers an alternative approach to photovoltaic devices for solar electricity generation. The water oxidation reaction (WOR) on the anode and oxygen reduction reaction (ORR) on the cathode is an ideal design for energy transfer owing to their superiority in terms of cleanliness, eco-friendliness, and natural abundance. However, solar electricity production based on O2 circulation by a fuel-free PEC cell is very challenging because it is extremely hard to extract electrons from water molecules owing to the uphill and sluggish WOR together with enormous overpotential for the cathodic ORR. Herein, a PEC cell based on the OH- /O2 redox pair is reported for efficient and sustainable solar electricity production by using two photoelectrodes of TiO2 and polyterthiophene in alkaline electrolyte. This fuel-free PEC cell delivers an open-circuit voltage up to 0.90 V and a maximum power density of 222 μW cm-2 with O2 -saturated NaOH electrolyte under AM 1.5 G solar irradiation. A record solar-to-electricity conversion efficiency of 0.22 % is achieved in the case of tandem illumination of the two photoelectrodes. In addition, the dual photoelectrode remains robust in accelerated and day-night cycling operation under natural atmosphere for more than a week. This PEC cell is free of fuel, separating membranes, and cocatalyst, which may guide future designs for clean and simple devices for solar energy conversion.

Chronic gouty arthritis, caused by a persistent increase in, and the deposition of, soluble uric acid (sUA), can induce pathological chondrocyte destruction; however, the effects of urate transport and intracellular sUA on chondrocyte functionality and viability are yet to be fully determined. Thus, the aim of the present study was to investigate the presence and functionality of a urate transport system in chondrocytes. The expression profiles of two primary urate reabsorptive transporters, glucose transporter 9 (GLUT9) and urate transporter 1 (URAT1), in human articular cartilage and chondrocyte cell lines were examined via western blotting, reverse transcription‑quantitative PCR, immunohistochemistry and immunofluorescence. Then, chondrocytes were incubated with exogenous sUA at increasing concentrations. Negative control assays were conducted via the specific knockdown of GLUT9 and URAT1 with lentiviral short hairpin (sh)RNAs, and by pretreatment with benzbromarone, a known inhibitor of the two transporters. Intracellular UA concentrations were measured using colorimetric assays. The expression levels of GLUT9 and URAT1 were determined in cartilage tissues and chondrocyte cell lines. Incubation of chondrocytes with sUA led to a concentration‑dependent increase in intracellular urate concentrations, which was inhibited by GLUT9 or URAT1 knockdown, or by benzbromarone pretreatment (27.13±2.70, 44.22±2.34 and 58.46±2.32% reduction, respectively). In particular, benzbromarone further decreased the already‑reduced intracellular UA concentrations in HC‑shGLUT9 and HC‑shURAT1 cells by 46.79±2.46 and 39.79±2.22%, respectively. Cells overexpressing GLUT9 and URAT1 were used as the positive cell control, which showed increased intracellular UA concentrations that could be reversed by treatment with benzbromarone. In conclusion, chondrocytes may possess an active UA transport system. GLUT9 and URAT1 functioned synergistically to transport UA into the chondrocyte cytoplasm, which was inhibited by specific gene knockdowns and drug‑induced inhibition. These results may be fundamental in the further investigation of the pathological changes to chondrocytes induced by sUA during gouty arthritis, and identified UA transport processes as potential targets for the early control of chronic gouty arthritis.

Photoelectrochemical (PEC) oxidation of glucose on bismuth vanadate (BiVO4) electrode for sensor application and solar-to-electricity production was investigated in this work. The BiVO4-based PEC sensor displays a sensitivity of 17.38 μA cm−2 mM−1 for glucose sensing in the concentration range of 5–35 mM. Furthermore, electricity can be collected by extracting solar energy and bioenergy in a certain photo fuel cell (PFC) device via anodic reaction of photo-oxidizing glucose and cathodic reaction of oxygen reduction. This study offers a promising strategy for glucose sensing and provides a cost-effective way for solar-to-electricity generation.

The building envelope is a key factor affecting the energy efficiency of buildings. Often, building envelope design requires both explicit knowledge and tacit knowledge. In practice, the tacit knowledge embedded in existing envelope design cases is significantly underused. To enhance the application of such knowledge, this paper proposes a case-based reasoning (CBR) model for the decision support of building envelop design during the preliminary design stage. A case library of 100 certificated green public and commercial buildings is used to develop the model. An experiment on a test case is performed to verify the methodology and usability of the model. Furthermore, records for 25 certificated green public buildings are used to validate the effectiveness of the model. The result shows that the accuracy rate of the CBR model is 84% when considering heating and cooling demands of envelopes. In conclusion, the proposed CBR model is promising for improving the efficiency of envelope design for public and commercial buildings while reducing the reliance on expert participation.

Electron transfer processes from semiconductor to molecular catalysts was studied in a model hybrid photocatalytic hydrogen evolution system composed of [Co((III))(dmgH)2PyCl] (CoPy) and CdS under different pH conditions. Thermodynamic and kinetic studies revealed that photocatalytic H2 evolution under high pH conditions (pH 13.5) can only account for the thermodynamically more favorable single-step simultaneous two-electron transfer from photoirradiated CdS to Co(III)Py to produce unavoidable intermediate Co(I)Py, rather than a two-step successive one-electron transfer process. This finding not only provides new insight into the charge transfer processes between semiconductors and molecular catalysts but also opens up a new avenue for the assembly and optimization of semiconductor-molecular catalyst hybrid systems processed through multielectron transfer processes.

The linear-array lidar has been recently developed and applied for its superiority of vertically non-scanning, large field of view, high sensitivity and high precision. The beam shaper is the key component for the linear-array detection. However, the traditional beam shaping approaches can hardly satisfy our requirement for obtaining unbiased and complete backscattered intensity data. The required beam distribution should roughly be oblate U-shaped rather than Gaussian or uniform. Thus, an optimal beam shaping approach is proposed in this paper. By employing a pair of conical lenses and a cylindrical lens behind the beam expander, the expanded Gaussian laser was shaped to a line-shaped beam whose intensity distribution is more consistent with the required distribution. To provide a better fit to the requirement, off-axis method is adopted. The design of the optimal beam shaping module is mathematically explained and the experimental verification of the module performance is also presented in this paper. The experimental results indicate that the optimal beam shaping approach can effectively correct the intensity image and provide ~30% gain of detection area over traditional approach, thus improving the imaging quality of linear-array lidar.

Biomarkers such as DNA, RNA, and protein are powerful tools in clinical diagnostics and therapeutic development for many diseases. Identifying RNA expression at the single cell level within the morphological context by RNA in situ hybridization provides a great deal of information on gene expression changes over conventional techniques that analyze bulk tissue, yet widespread use of this technique in the clinical setting has been hampered by the dearth of automated RNA ISH assays. Here we present an automated version of the RNA ISH technology RNAscope that is adaptable to multiple automation platforms. The automated RNAscope assay yields a high signal‐to‐noise ratio with little to no background staining and results comparable to the manual assay. In addition, the automated duplex RNAscope assay was able to detect two biomarkers simultaneously. Lastly, assay consistency and reproducibility were confirmed by quantification of TATA‐box binding protein (TBP) mRNA signals across multiple lots and multiple experiments. Taken together, the data presented in this study demonstrate that the automated RNAscope technology is a high performance RNA ISH assay with broad applicability in biomarker research and diagnostic assay development. J. Cell. Biochem. 117: 2201–2208, 2016. © 2016 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc.

A young female with recurrent tophaceous gout and infertility presented to our clinic. On clinical evaluation, hypoglycaemia, hypertriglyceridaemia, lactic acidosis, and hepatomegaly were noted. Targeted gene sequencing revealed a novel composite heterozygous c.190G>T/c.508C>T mutation in the G6PC gene of the patient, leading to a diagnosis of glycogen storage disease type Ia. Her father possessed a heterozygous c.190G>T mutation, and her mother possessed a heterozygous c.508C>T mutation. A search of the previous literature revealed 16 reported cases of glycogen storage disease type Ia with gout. Here, we describe a female patient with gout, review previous cases, and discuss the mechanisms of gout and hyperuricaemia in glycogen storage disease type Ia.

Photocatalytic selective oxidation represents an environment-friendly strategy for chemical transformation. Herein, we report the oxidation of sulfides into sulfoxides with high selectivity (up to 99%) on Pt/BiVO 4 photocatalyst in water under visible light illumination. The system exhibited excellent performance for the oxidation of sulfides in water compared with organic solvents. Isotopic oxygen (H 2 18 O) experiments clearly revealed that the oxygen atoms in the sulfoxide were mainly from water. Electron paramagnetic resonance demonstrated that the reactive oxygen species were the peroxy species, which were generated from water oxidation on BiVO 4 via the two-electron pathway. Dioxygen was reduced to water through proton-coupled electron transfer.

Spatially resolved gene expression has emerged as a crucial technique to understand complex multicellular interactions within the tumor and its microenvironment. Interrogation of complex cellular interactions within the tumor microenvironment (TME) requires a multi-omics approach where multiple RNA and protein targets can be visualized within the same tumor sample and be feasible in FFPE sample types. Simultaneous detection of RNA and protein can reveal cellular sources of secreted proteins, identify specific cell types, and visualize the spatial organization of cells within the tissue. Examination of RNA by in situ hybridization (ISH) and protein by immunohistochemistry (IHC) or immunofluorescence (IF) are widely used and accepted techniques for the detection of biomarkers in tumor samples. Given the similarities in workflow, co-detection of RNA and protein by combining ISH and IHC/IF in a single assay can be a powerful multi-omics solution for interrogating the complex tumor and its microenvironment. In this report we combined the single cell, single molecule RNA ISH technology known as RNAscope with IHC/IF to simultaneously detect RNA and protein in the same FFPE tumor section using both chromogenic and fluorescence detection methods. We demonstrate co-localization of target mRNA and the corresponding protein in human cancer samples, visualize infiltration of immune cells into the TME, characterize the activation state of immune cells in the TME, identify single cell gene expression within cellular boundaries demarcated by IHC/IF, examine cell type-specific expression of multiple immune checkpoint markers, and distinguish endogenous T cells from activated CAR+ T cells. Overall, we show that co-detection of RNA by the RNAscope ISH assay and protein by the IHC/IF assay in the same FFPE section is a feasible methodology. The combined RNAscope ISH-IHC/IF workflow is a powerful technique that can be used to study gene expression signatures at the RNA and protein level with spatial and single cell resolution. By leveraging the strength of the similar workflows of RNAscope ISH and IHC/IF assays, this methodology combines transcriptomics and proteomics in the same tissue section, providing a multi-omics approach for characterizing complex tissues and revealing cell type specific gene expression with spatial and single cell resolution. Citation Format: Anushka Dikshit, Jyoti Phatak, Siobhan Kernag, Helly Pimental, Hailing Zong, Courtney Todorov, Lydia Hernandez, Jeffrey Kim, Bingqing Zhang, Courtney Anderson, Xiao-Jun Ma. Spatially resolve RNA and protein simultaneously in FFPE tumor samples by combining RNAscope in situ hybridization and immunohistochemistry assays [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2706.

About 25% of non-small cell lung cancer (NSCLC) patients bear one or more KRAS mutations in their tumors, which is correlated with poor prognosis. The precise identification of somatic mutations in tumors is becoming increasingly important for studying tumor progression and developing targeted therapies. While sequencing technologies allow for mutation-profiling, they do not permit direct visualization and association of genetic alterations with cellular morphology. In addition, DNA mutational status does not predict expression of the mutant allele which may provide information connecting genotype to phenotype. Therefore, a technology for mutation detection at the transcript level directly in the tumor context is desirable. To address this need we developed a specialized RNA in situ hybridization (ISH) method known as BaseScope. The BaseScope assay has a unique signal amplification system that allows for highly sensitive and specific detection of single nucleotide point mutations in tissues. BaseScope probes specific for KRAS G12C, G12A, G12V, G12S and wild type KRAS were designed and expression of each point mutation was assessed in a NSCLC tumor microarray with 48 tumor cores with known KRAS mutation status as determined by DNA sequencing. RNA quality and background signal threshold for each tumor core were determined using PPIB (positive) and dapB (negative) control probes. Using the sequencing data as the gold standard, the BaseScope assay demonstrated 83-100% sensitivity and 97-100% specificity for various KRAS mutations [Table 1]. For KRAS G12C, the assay correctly identified all 6 sequencing-positive cores and identified the rest as negatives. For KRAS G12V, the assay detected 5 of 6 mutated cores with 100% specificity. Interestingly, for KRAS G12S and KRAS G12A mutations, the BaseScope assay demonstrated 100% sensitivity and 97% specificity. Furthermore, it was observed that 100% of the KRAS-mutated tumors showed expression for both wild type and mutant KRAS alleles within these NSCLC tumors. In summary, we demonstrate the development of an RNA ISH assay for point mutations detection with morphological context in FFPE tissues. Unlike current sequencing methods that lack spatial information this assay has the unique ability to identify very small subclones whose frequency within the tumor might fall below the detection limit of sequencing. Performance characteristics of BaseScope KRAS assaysKRAS POINT MUTATIONSNo. of cores with specified mutationsNo. of cores without specified mutationsBaseScope sensitivityBaseScope specificityG12C633100% (6/6)100% (33/33)G12A336100% (3/3)97.2% (35/36)G12V63383% (5/6)100% (33/33)G12S138100% (1/1)97.3% (37/38) Citation Format: Anushka Dikshit, Helen Jarnagin, Emerald Doolittle, Courtney Anderson, Bingqing Zhang, Xiao-Jun Ma. Visualization of KRAS point mutations in non-small cell lung cancer tumors with morphological context using the BaseScope in situ hybridization assay [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2705.

(Photo)electrochemistry enables the synthesis of high-value fine chemicals and highly selective activation of molecules that are difficult to prepare using conventional chemical methods. In this work, light-driven NADH (reduced nicotinamide adenine dinucleotide) regeneration is achieved using a molecular Rh(III) mediator on Si photoelectrodes. This process is observed to be highly sensitive to the surface nature of Si photoelectrodes, exhibiting an overpotential reduction up to 600 mV on Si nanowires (SiNWs) as compared to planar Si. The use of a molecular mediator and SiNWs enables 100% selectivity toward NADH synthesis within a broad potential window. The origin of the striking difference is identified as the multifaceted nature of SiNWs.

As we strive to prolong patient survival, the advent of targeted therapy for the treatment of ovarian cancer has significantly added to our armamentarium. Unfortunately, both chemotherapy and molecularly targeted PARPi approaches share the overarching limitation of the emergence of drug resistance. One key aspect towards realizing the potential of targeted therapies is a better understanding of the intrinsic and acquired resistance mechanisms that limit their efficacy. Through comprehensive genomic analysis of post-treatment patient samples, we recently identified the most common mechanism of acquired drug resistance in high-grade serous ovarian cancer (HGSC) to date, a transcriptional fusion involving ABCB1. ABCB1 encodes P-gp also known as multidrug resistance protein 1 (MDR1), a multi-transmembrane domain protein that is a member of the superfamily of ATP binding cassette (ABC) transporters involved in the cellular efflux of chemotherapeutic drugs. The SLC25A40-ABCB1 fusion was associated with upregulation of ABCB1 expression, whilst leaving the predicted ABCB1 protein unaltered. Interestingly, fusion events were only detected in patients who had been exposed to chemotherapies that are known substrates of P-gp, with the probability of fusion events closely correlated to the number of lines of P-gp substrate chemotherapy. Surprisingly, WGS analysis of patient samples revealed that not all tumor cells in fusion-positive patients carry the fusion. An intriguing possibility is that resistance within tumor sites is spatially ordered rather than random. Identifying such patterning could explain why tumor eradication has not been effective for the majority of HGSC patients to date. To address the subclonal localization and spatial patterning of ABCB1 fusions in HGSC, CASCADE (rapid autopsy program), biopsy specimens, and PDX tissue are being prescreened (qRT-PCR) to identify those with the highest levels of ABCB1 expression and thus most likely to harbor fusions. In situ DNA and RNA detection assays are being employed to identify ABCB1 fusions. To examine whether fusion negative cells also overexpress P-gp, IHC analysis will then be conducted to co-register fusion positivity and protein expression. We have successfully identified fusion events using ACD BaseScope technology and examined the localization of key HGSC genes including CCNE1 and ABCB1 through PCR FISH assays. In summary, this study will decipher the diversity of resistance mechanisms within individual HGSC patients, thereby providing critical information required for next-generation chemotherapy and PARPi clinical trials aimed at reversing or bypassing acquired resistance. Citation Format: Kathleen I. Pishas, Elizabeth L. Christie, Jessica A. Beach, Kathryn Alsop, Alison Freimund, Nidhi Vashistha, Niyati Jhaveri, Emerald Doolittle, Wei Wei, Bingqing Zhang, Xiao-Jun Ma, David D.L. Bowtell. Spatial characterization of drug resistance in ovarian cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr A65.

Assessing the environmental impact (EI) of regulatory planning (a lower tier in the Chinese urban planning system) has significant implications for improving the environmental performance of the future built environment. Current environmental impact assessments generally focus on microscale and macroscale, while mesoscale studies at region level are lacking. To fill this gap, this paper proposes an LCA-based environmental impact assessment model for regulatory planning, which focuses on embodied EI of urban construction and operational EI associated with the operation of the built environment. To ensure the regulatory plans both meet the environmental limits and fit well with environmental policy requirements, this paper also presents a new impact analysis method by integrating environmental carrying capacity analysis and distance to target method. The Statutory Plan of Shenzhen Futian Free Trade Zone 03-T2 is used as a case study to test and demonstrate the applying process and functions of the proposed method. Results indicate that the proposed model can effectively quantify the future environmental impact associated with a regional development plan, and can potentially be used as a tool for urban administrators to make decisions on regulatory planning revision and provide references for environmental management.

Professional antigen presenting cells such as dendritic cells (DCs) are vital for priming naive CD8+ T cells and developing a sustainable anti-tumor immune response. Natural killer (NK) cells within the tumor microenvironment (TME) recruit a specific population of DCs called conventional type 1 DCs (cDC1s). However, these cells are low in abundance making their detection in the tissue context challenging. To interrogate the presence of cDC1 and NK cells in the TME and reveal their spatial relationship to each other we utilized the highly sensitive and specific RNAscope Multiplex Fluorescence in situ hybridization (ISH) assay. NK cells and cDC1 cells were identified by using cell specific marker probes in 4 cervical cancer samples. Similarly, CTLs were visualized to determine if there is a correlation between the presence of cDC1 and NK cells and infiltration of CTLs within the cervical cancer tumors. Our results revealed a strong correlation between the presence of NK cells, cDC1 cells, and CTLs within 3 out of 4 cervical cancer samples. The NK cells showed expression of the chemokines XCL1 and CCL5, suggesting that the XCR1+/CCR5+ cDC1 cells may have been potentially recruited by these NK cells in the TME. Regions high in cDC1 and NK cells also showed significantly higher levels of CTL recruitment, indicated by the presence of CD8+/IFNG+ T cells. Conversely, 1 of the 4 cervical cancer samples demonstrated relatively lower levels of NK cells which correlated with lower cDC1 cells and in turn lower CTL infiltration. In conclusion, by utilizing the RNAscope Multiplex ISH assay we identified and visualized the spatial relationship between NK cells, CTLs, and cDC1 cells, highlighting the strength of this technology to spatially interrogate the TME.

Photoelectrochemical (PEC) glucose sensing has been developed as a novel promising electrochemical approach for advanced analysis due to the advantages of high sensitivity and selectivity as well as good stability. Here, we present a Fe2O3 nanorod (Fe2O3NR)-based PEC sensor, which shows excellent performance in glucose detection for the concentrations in the range of 0.2–2.0 mM with a linear correlation coefficient of 0.997 and a sensitivity of 100.46 μA cm−2 mM−1. Furthermore, the glucose determination can be achieved without inputting external electrical power via a photo fuel cell (PFC) which is fabricated by the Fe2O3NR photoanode and platinum cathode. The generated maximum power density (Pmax) shows a linear sensitivity of 3.53 μW cm−2 mM−1 to the variation of glucose concentrations from 0.5 to 2.5 mM with correlation coefficient of 0.990. Therefore, the Fe2O3NR-based PEC sensor not only realizes the electricity generation from renewable solar energy and bioenergy but also accomplishes the detection of glucose by self-power.

Because precision medicine is highly dependent on the accurate detection of biomarkers, there is an increasing need for standardized and robust technologies that measure RNA biomarkers in situ in clinical specimens. While grind-and-bind assays like RNAseq and quantitative RT-PCR enable highly sensitive gene expression measurements, they also require RNA extraction and thus prevent valuable expression analysis within the morphological tissue context. The in situ hybridization (ISH) assay described here can detect RNA target sequences as short as 50 nucleotides at single-nucleotide resolution and at the single-cell level. This assay is complementary to the previously developed commercial assay and enables sensitive and specific in situ detection of splice variants, short targets, and point mutations within the tissue. In this protocol, probes were designed to target unique exon junctions for two clinically important splice variants, EGFRvIII and METΔ14. The detection of short target sequences was demonstrated by the specific detection of CDR3 sequences of T-cell receptors α and β in the Jurkat T-cell line. Also shown is the utility of this ISH assay for the distinction of RNA target sequences at single-nucleotide resolution (point mutations) through the visualization of EGFR L858R and KRAS G12A single-nucleotide variations in cell lines using automated staining platforms. In summary, the protocol shows a specialized RNA ISH assay that enables the detection of splice variants, short sequences, and mutations in situ for manual performance and on automated stainers.

Because precision medicine is highly dependent on the accurate detection of biomarkers, there is an increasing need for standardized and robust technologies that measure RNA biomarkers in situ in clinical specimens. While grind-and-bind assays like RNAseq and quantitative RT-PCR enable highly sensitive gene expression measurements, they also require RNA extraction and thus prevent valuable expression analysis within the morphological tissue context. The in situ hybridization (ISH) assay described here can detect RNA target sequences as short as 50 nucleotides at single-nucleotide resolution and at the single-cell level. This assay is complementary to the previously developed commercial assay and enables sensitive and specific in situ detection of splice variants, short targets, and point mutations within the tissue. In this protocol, probes were designed to target unique exon junctions for two clinically important splice variants, EGFRvIII and METΔ14. The detection of short target sequences was demonstrated by the specific detection of CDR3 sequences of T-cell receptors α and β in the Jurkat T-cell line. Also shown is the utility of this ISH assay for the distinction of RNA target sequences at single-nucleotide resolution (point mutations) through the visualization of EGFR L858R and KRAS G12A single-nucleotide variations in cell lines using automated staining platforms. In summary, the protocol shows a specialized RNA ISH assay that enables the detection of splice variants, short sequences, and mutations in situ for manual performance and on automated stainers.

This Statistical Analysis Plan details the statistical procedures to be applied for the analysis of data for the multicenter electroencephalography study. It consists of a basic description of the study in broad terms and separate sections that detail the methods of different aspects of the statistical analysis, summarized under the following headings (a) Background; (b) Definitions of protocol violations; (c) Definitions of objectives and other terms; (d) Variables for analyses; (e) Handling of missing data and study bias; (f) Statistical analysis of the primary and secondary study outcomes; (g) Reporting of study results; and (h) References. It serves as a template for researchers interested in writing a Statistical Analysis Plan.

Solar water splitting promises a solution to challenges associated with the intermittent nature of solar energy. Of different implementations, photoelectrochemical water splitting, where one or more photoelectrodes harvest light and catalyze water splitting, represents a convenient platform to understand the governing principles of charge behaviors, especially at the light absorber|H2O interface. This Perspective recognizes and discusses the importance of the photoelectrode surface to solar water splitting performance. It presents discussions within the context of a prototypical water splitting material, Ta3N5, which has gained growing attention lately for its outstanding initial performance. Insights into the mechanisms by which Ta3N5 functions are presented, followed by examples of recent efforts to circumvent the issues that Ta3N5 decays rapidly under solar water splitting conditions. Our visions on the future directions of semiconductor-based solar water splitting will be presented at the end.