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Supplemental Table 1, NMR Results, Supplemental Table 2, Supplemental Figures 1 and 2

Introduction: The pandemic caused by coronavirus disease 2019 has adversely affected the health of all individuals including pregnant women. The susceptibility of pregnant women and their foetuses to severe acute respiratory syndrome coronavirus 2 infection is not clearly known. The objective of this review is to summarise the available evidence on foetal and neonatal outcomes of pregnant women with confirmed coronavirus disease 2019. Methods: The review was conduced by searching the PubMed and MedRxiv databases for studies reporting outcome of pregnancy with confirmed coronavirus disease from December 2019 to April 2020 using combination of terms "pregnancy", "coronavirus disease 2019", "foetal outcome" and "neonatal outcome". Only peer reviewed articles reporting outcome of pregnancy with confirmed coronavirus disease 2019 were included in the review irrespective of language. Ongoing pregnancies, induced abortion, and pregnancies without outcome were excluded from analysis. Variables extracted and analysed included gestational age at delivery, mode of delivery, foetal and neonatal outcome. Results: In total, 21 studies consisting 230 pregnant women including three twin pregnancies were enrolled in the study. Fever and cough were the most common symptoms reported in pregnant women. One hundred and sixty-two (70.4%) pregnant women underwent cesarean section and there were 68 (29.6%) preterm deliveries. Eight babies tested positive out of 161 newborns that were tested for coronavirus infection. Among adverse outcomes reported, there were two miscarriages, two still births and only one neonatal death. Conclusions: Outcome of pregnancy with coronavirus disease 2019 in late trimester appears to be favourable. Occurrence of preterm delivery and cesarean section appear higher among infected pregnant women in comparison to general population. There was no conclusive evidence of vertical transmission.

Introduction: Blood transfusion is a common practice among children with severe anaemia. A common assumption among physician is that transfusion of 5 ml/kg packed red cell will increase the haemoglobin of the recipient by 1 gm/dl. This study was aimed at assessing the effect of volume of blood product and hematocrit of donor blood in the rise of haemoglobin concentration after transfusion in children without active bleeding. Methods: A prospective observational study was conducted in 32 children aged between one to 15 years without active bleeding who received blood transfusion in Paediatric ward and Paediatric Intensive Care Unit of a tertiary care hospital in Nepal from December 2013 to November 2014. Haemoglobin levels before and after transfusion were compared and any adverse reactions that occurred during transfusion were recorded. Hematocrit of each donor blood was measured. Results: The overall rise of haemoglobin after transfusion of blood product with hematocrit of 53 ± 8.9 % at the rate 14.5 ± 6.0 ml/kg was 2.7 ± 1.4 gm/dl. The rise of haemoglobin after transfusion of whole blood with hematocrit of 38.8 ± 4.4% at the rate 15.5 ± 6.9 ml/kg was 1.8 ± 1.2 gm/dl. The rise of haemoglobin after transfusion of packed cell with hematocrit 57.0 ± 4.8 % at the rate 14.2 ± 5.8 ml/kg was 2.9 ± 1.4 gm/dl. Conclusion: This study supports the general consensus of 1 gm/dl rise of haemoglobin after transfusion of packed red cell at 5 ml/kg.

Urban areas provide a range of benefits to sustain human livelihood and contribute to human well-being through urban ecosystem services. Open spaces in core urban areas of Kathmandu valley in Nepal carries multiple advantages of stimulating social cohesion, offers safe area immediately after a crisis induced by natural hazards, contributes in environmental improvement and mitigates urban flooding. In most urban areas of Nepal, unplanned urbanization has resulted in alteration of landscapes from permeable vegetated surfaces to a series of impervious interconnected surfaces resulting in large quantities of stormwater runoff, requiring wider implementation of water sensitive urban design. After 2015 Gorkha earthquake, several blue-green infrastructure projects are implemented by local governments inside Kathmandu valley in open spaces. This paper presents application examples of green infrastructure projects and through case studies provides a framework for optimization of green infrastructure systems in Nepal. The paper also provides a practitioners perspective on the current state of knowledge, highlights technical challenges in green infrastructure implementation in Kathmandu and points out recommendations to overcome them.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer with poor prognosis and chemotherapy with gemcitabine has limited effects and is associated with development of drug resistance. Treatment of Panc1 and MiaPaca2 pancreatic cancer cells with gemcitabine induced expression of the orphan nuclear receptor 4A2 (NURR1) and analysis of The Cancer Genome Atlas indicated the NURR1 is overexpressed in pancreatic tumors and is a negative prognostic factor for patient survival. Results of NURR1 knockdown or treatment with the NURR1 antagonist 1,1-bis(3΄-indolyl)-1-(p-chlorophenyl)methane (C-DIM 12) demonstrated that NURR1 was prooncogenic in pancreatic cancer cells and regulated cancer cell and tumor growth and survival. NURR1 is induced by gemcitabine and serves as a key drug resistance factor and is also required for gemcitabine-induced cytoprotective autophagy. NURR1-regulated genes were determined by RNA sequencing of mRNAs expressed in MiaPaCa2 cells expressing NURR1 and in CRISPR/Cas9 gene–edited cells for NURR1 knockdown and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of the differentially expressed genes showed that autophagy was the major pathway regulated by NURR1. Moreover, NURR1 regulated expression of two major autophagic genes, ATG7 and ATG12, which are also overexpressed in pancreatic tumors and like NURR1 are negative prognostic factors for patient survival. Thus, gemcitabine-induced cytoprotective autophagy is due to the NURR1–ATG7/ATG12 axis and this can be targeted and disrupted by NURR1 antagonist C-DIM12 demonstrating the potential clinical applications for combination therapies with gemcitabine and NURR1 antagonists. Significance: Gemcitabine induces NURR1-dependent ATG7 and ATG12 cytoprotective autophagy in PDA cells that can be reversed by NURR1 antagonists.

Paraspeckles compound 1 (PSPC1) is a multifunctional protein that plays an important role in cancer cells where PSPC1 is a master regulator of pro-oncogenic responses that includes activation of TGFβ (TGFβ1) and TGFβ-dependent EMT and metastasis. The pro-oncogenic activities of PSPC1 closely resembled those observed for the orphan nuclear receptor 4A1 (NR4A1, Nur77) and knockdown of NR4A1 decreased expression of PSPC1 in MDA-MB-231 breast, H1299 lung and SNU449 liver cancer cells. Similar results were observed in these same cell lines after treatment with bis-indole – derived (CDIMs) NR4A1 antagonists. Moreover, PSPC1-dependent regulation of TGFβ, genes associated with cancer stem cells and epithelial to mesenchymal transition (EMT) were also downregulated after NR4A1 silencing or treatment of breast, lung and liver cancer cells with CDIM/NR4A1 antagonists. Results of chromatin immunoprecipitation (ChIP) assays suggest that NR4A1 regulates PSPC1 through interaction with an NBRE sequence in the PSPC1 gene promoter. These results coupled with in vivo studies showing that NR4A1 antagonists inhibit breast tumor growth and downregulate PSPC1 in tumors indicate that the pro-oncogenic nuclear PSPC1 factor can be targeted by CDIM/NR4A1 antagonists.

Background and objective The prevalence of non-alcoholic fatty liver disease (NAFLD) is 60% in patients with type 2 diabetes mellitus (T2DM). NAFLD can lead to non-alcoholic steatohepatitis (NASH), both of which are the leading causes of cirrhosis. This study was undertaken to evaluate whether empagliflozin, a sodium-glucose cotransporter-2 (SGLT-2) inhibitor, reduces liver fat content in these patients after therapy. Methods After enrolling patients of T2DM with NAFLD, they were administered empagliflozin 10 mg once daily orally for six months without modifying existing oral hypoglycemic agents (OHA) if any. All demographic data were collected, and anthropometric measurements, as well as laboratory investigations, were performed, and controlled attenuation parameter (CAP) and liver stiffness (LS) were measured using FibroScan® (Echosens, Paris, France) at baseline, and six months of therapy. The adverse effects related to therapy were also taken into account. Results There was a significant decrease in mean CAP value from 282.07 ± 47.29 dB/m to 263.07 ± 49.93 dB/m and LS from 5.89 ± 4.23 kPa to 5.04 ± 1.49 kPa along with a significant decrease in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transferase (GGT) among the patients. Compared to the baseline, there was a significant reduction in post-treatment weight, body mass index (BMI), and blood pressure (BP). The most commonly observed adverse effects of the therapy were urinary tract infection (UTI) (17.8%), nasopharyngitis (11.9%), and hypoglycemia (10.71%). Conclusion A reduction in hepatic fat content was seen in our prospective study cohort after six months of empagliflozin therapy. Empagliflozin also led to beneficial effects such as weight loss and reduction in transaminases and GGT. Given the absence of significant side effects of the therapy, empagliflozin could be used as an effective treatment modality for T2DM patients with NAFLD, which are two conditions commonly seen in combination.

Background Assessment of nonalcoholic fatty liver disease (NAFLD) includes estimation of liver fat (steatosis). Controlled attenuation parameter (CAP) value obtained by FibroScan® (Echosens, Paris, France) is an alternative to liver biopsy for diagnosing and estimating steatosis (S). This study aimed to estimate the liver fat by CAP in NAFLD patients. Methods An observational cross-sectional study was conducted at the Liver Unit of Bir Hospital, from January 2021 to May 2021 after ethical clearance from the Institutional Review Board of the National Academy of Medical Sciences. A convenient sampling method was used. Data were analyzed with descriptive and inferential statistics involving bivariate and multivariate analysis. Results A total of 127 NAFLD patients were enrolled. The mean (±SD) CAP value was 271.53 (±50.69) dB/m. Total cholesterol, triglyceride, and body mass index (BMI) correlated positively (p

The ACT‐America project is a NASA Earth Venture Suborbital‐2 mission designed to study the transport and fluxes of greenhouse gases. The open and freely available ACT‐America data sets provide airborne in situ measurements of atmospheric carbon dioxide, methane, trace gases, aerosols, clouds, and meteorological properties, airborne remote sensing measurements of aerosol backscatter, atmospheric boundary layer height and columnar content of atmospheric carbon dioxide, tower‐based measurements, and modeled atmospheric mole fractions and regional carbon fluxes of greenhouse gases over the Central and Eastern United States. We conducted 121 research flights during five campaigns in four seasons during 2016–2019 over three regions of the US (Mid‐Atlantic, Midwest and South) using two NASA research aircraft (B‐200 and C‐130). We performed three flight patterns (fair weather, frontal crossings, and OCO‐2 underflights) and collected more than 1,140 h of airborne measurements via level‐leg flights in the atmospheric boundary layer, lower, and upper free troposphere and vertical profiles spanning these altitudes. We also merged various airborne in situ measurements onto a common standard sampling interval, which brings coherence to the data, creates geolocated data products, and makes it much easier for the users to perform holistic analysis of the ACT‐America data products. Here, we report on detailed information of data sets collected, the workflow for data sets including storage and processing of the quality controlled and quality assured harmonized observations, and their archival and formatting for users. Finally, we provide some important information on the dissemination of data products including metadata and highlights of applications of ACT‐America data sets., Key Points Atmospheric Carbon and Transport – America (ACT‐America) provides a unique, weather‐oriented collection of atmospheric CO2, CH4, trace gases, and meteorological properties measurementsACT‐America data are free and open to the public from the Oak Ridge National Laboratory Distributed Active Archive Center (ORNL DAAC)ACT‐America data are uniquely suited to improve the accuracy and precision of regional inverse greenhouse gas (GHG) flux estimates

Nuclear receptor 4A1 (NR4A1, Nur77, TR3) is more highly expressed in breast and solid tumors compared to non-tumor tissues and is a pro-oncogenic factor in solid tumor-derived cancers. NR4A1 regulates cancer cell growth, survival, migration, and invasion, and bis-indole-derived compounds (CDIMs) that bind NR4A1 act as antagonists and inhibit tumor growth. Preliminary structure-binding studies identified 1,1-bis(3′-indolyl)-1-(3,5-disubstitutedphenyl)methane analogs as NR4A1 ligands with low KD values, we further investigated the anticancer activity of the four most active analogs (KD’s ≤ 3.1 µM) in breast cancer cells and in athymic mouse xenograft models. The treatment of MDA-MB-231 and SKBR3 breast cancer cells with the 3-bromo-5-methoxy, 3-chloro-5-trifluoromethoxy, 3-chloro-5-trifluoromethyl, and 3-bromo-5-trifluoromethoxy phenyl-substituted analogs decreased cell growth and the expression of epidermal of growth factor receptor (EGFR), hepatocyte growth factor receptor (cMET), and PD-L1 as well as inhibited mTOR phosphorylation. In addition, all four compounds inhibited tumor growth in athymic nude mice bearing MDA-MB-231 cells (orthotopic) at a dose of 1 mg/kg/d, which was not accompanied by changes in body weight. These 3,5-disubstituted analogs were the most potent CDIM/NR4A1 ligands reported and are being further developed for clinical applications.

The nuclear receptor (NR) superfamily of transcription factors encodes expression of 48 human genes that are important for maintaining cellular homeostasis and in pathophysiology, and this has been observed for all sub-families including orphan receptors for which endogenous ligands have not yet been identified. The orphan NR4A1 (Nur77 and TR3) and other members of this sub-family (NR4A2 and NR4A3) are immediate early genes induced by diverse stressors, and these receptors play an important role in the immune function and are up-regulated in some inflammatory diseases including solid tumors. Although endogenous ligands for NR4A have not been identified, several different classes of compounds have been characterized as NR4A1 ligands that bind the receptor. These compounds include cytosporone B and structurally related analogs, bis-indole derived (CDIM) compounds, the triterpenoid celastrol and a number of other chemicals including polyunsaturated fatty acids. NR4A1 ligands bind different regions/surfaces of NR4A1 and exhibit selective NR4A1 modulator (SNR4AM) activities that are dependent on ligand structure and cell/tissue context. NR4A1 ligands exhibit pharmacologic activities in studies on cancer, endometriosis metabolic and inflammatory diseases and are promising agents with clinical potential for treating multiple diseases.

Colorectal cancer (CRC) is a highly complex disease with multiple risk factors. The orphan nuclear receptor 4A1 (NR4A1) is overexpressed in several cancers and is a negative prognostic factor for cancer patient survival. Previous reports indicate a potential role for overexpression of NR4A1 in T-cell exhaustion and in this study, we aim to investigate the antitumorigenic activity of two bis-indole derived ligands (DIMs) that act as receptor antagonists. Immune competent C57BL/6 mice and mouse MC-38 colon cancer cells were used and tumor Infiltrating Lymphocytes (TILs) were isolated from mice either untreated or treated with CDIM/NR4A1 antagonists. FACS analysis and Real -Time PCR were performed to determine expression of exhaustion markers in these tumor T-cell population. 1,1-Bis(3΄-indolyl)-1-(3-bromo-5-trifluoromethoxyphenyl)methane (DIM-3-Br-5-OCF3) and the 3,5-dichlorophenyl analog (DIM-3,5-Cl2) at doses of 2.5 and 7.5 mg/kg/d inhibited tumor growth and downregulated expression of PD-L1, an NR4A1-regulated gene in MC-38 - derived tumors and cells. The mouse MC-38 colon cancer cell line was used in a syngeneic mouse model of colon cancer and tumor infiltrating lymphocytes (TILs) from MC-38 cell-derived colon tumors exhibited multiple markers of T-cell exhaustion in both CD8+ and CD4+ T-cells. Lymphocytes from an enlarged spleen in these tumor-bearing animals also exhibited markers of CD8+ and CD4+ T-cell exhaustion. Analysis of CD8+ cells from TILs showed that treatment with the NR4A1 antagonists modulated expression of several genes associated with T-cell exhaustion namely a decrease in TOX, TOX2 and NFAT mRNAs, activation of T-Bet. The percentage of CD8+ and CD4+ T-cells from tumors and spleen expressing PD-1, 2B4 and TIGIT was decreased in the treated vs control mice and TIM3 expression was also decreased in CD8+ (tumors and spleen) and CD4+ (tumors) T-cells. These findings suggest that NR4A1 antagonists are highly effective as anticancer agents in this mouse syngeneic colon tumor model by inactivating the pro-oncogenic activities of NR4A1 in tumors and by remediating NR4A1-regulated T-cell exhaustion in tumor and splenic lymphocytes. Citation Format: Kumaravel Mohankumar, Gus Wright, Subhashree Kumaravel, Rupesh Shrestha, Lei Zhang, Maen Abdelrahim, Robert S. Chapkin, Stephen Safe. Nuclear receptor 4A1 ligands target T-cell exhaustion in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 236.

BackgroundColorectal cancer (CRC) is a highly complex disease with multiple risk factors and both genetic and environmental components contribute to disease incidence.1 2 Cancer immunotherapy using immune-checkpoint blockades represents a major advance in treatment strategy.3 4 The orphan nuclear receptor 4A1 (NR4A1) is overexpressed in lung, colon, liver and breast cancers and in Rhabdomyosarcoma and is a negative prognostic factor for cancer patient survival.5–8 Previous studies in breast cancer cells showed that PD-L1 was regulated by NR4A1 which activates transcription factor Sp1 bound to the PD-L1 gene promoter. Genome-wide studies have identified NR4A1 as a key mediator of T-cell dysfunction and NR4A1 also plays an important role in regulating genes which are involved in tumor-induced T-cell exhaustion.9 Bis-indole derived NR4A1 ligand that act as receptor antagonists have been developed in this laboratory and these compounds block pro-oncogenic NR4A1-regulated genes/pathways.MethodsImmune competent C57BL/6 mice and mouse MC-38 colon cancer cells were used and tumor Infiltrating Lymphocytes (TILs) were isolated from mice either untreated or treated with CDIM/NR4A1 antagonists. FACS analysis and Real -Time PCR were performed to determine expression of exhaustion markers in these tumor T-cell population.ResultsTwo compounds: 1,1-bis(3′-indolyl)-1-(3-bromo-5-trifluoromethoxyphenyl)methane (DIM-3-Br-5-OCF3) and 3,5-dichlorophenyl analog (DIM-3,5-Cl2) inhibited tumor growth and weight at doses of 2.5 and 7.5mg/kg/day (figure 1). Tumor CD8+ T-cells isolated from mice treated with DIM-3,5-Cl2 and DIM-3-Br-5-OCF3 exhibited decreased mRNA expression of NR4A1 and high mobility group – box transcription factors NFAT, TOX and TOX2 and increased mRNA levels of Interferon-γ (IFNγ), granzyme β (GzB) and perforin compared to control animals (figure 2). As TOX and TOX2 cooperate with NR4A1 to modulate CD8+ T-cell exhaustion, we investigated the expression of several inhibitory receptors of T-cell exhaustion in CD8+ TILs, including PD-1, 2B4, TIGIT and TIM3. Following treatment with DIM-3,5-Cl2 or DIM-3-Br-5-OCF3, there was a significant decrease in the percentage of PD1 and 2B4 cells and a decrease in TIGIT and TIM3 (figure 3). These results indicate that NR4A1 antagonists reverses T-cell exhaustion.ConclusionsNR4A1 plays a critical role in T-cell dysfunction, and this includes T-cell exhaustion.10 11 Our results demonstrate that the NR4A1 antagonists reverse many markers of T-cell exhaustion including activation of cytokines. The combined effects of NR4A1 antagonists in both tumors and T-cells result in inhibition of colon tumorigenesis by targeting pathways/genes in tumor cells and by enhancing immune surveillance via reversal of T-cell exhaustion.ReferencesAhmed M. Colon cancer: a Clinician's perspective in 2019. Gastroenterology Res 2020;13(1):1–10. Epub 2020/02/26. doi: 10.14740/gr1239.Keum N, Giovannucci E. Global burden of colorectal cancer: emerging trends, risk factors and prevention strategies. Nat Rev Gastroenterol Hepatol 2019;16(12):713–32. Epub 2019/08/29. doi: 10.1038/s41575-019-0189-8.Pardoll DM. The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 2012;12(4):252–64. Epub 2012/03/23. doi: 10.1038/nrc3239.Ribas A, Wolchok JD. Cancer immunotherapy using checkpoint blockade. Science 2018;359(6382):1350–1355.Yang et al. Distinct epigenetic features of tumor-reactive CD8+ T cells in colorectal cancer patients revealed by genome-wide DNA methylation analysis. Genome Biol 2019;21(1):2. doi: 10.1186/s13059-019-1921-y.Safe S, Karki K. The paradoxical roles of orphan nuclear receptor 4A (NR4A) in cancer. Mol Can Res 2020. Epub 2020/10/28. doi: 10.1158/1541-7786.MCR-20-0707.Lee SO, et al. Diindolylmethane analogs bind NR4A1 and are NR4A1 antagonists in colon cancer cells. Molecular Endocrinology. 2014;28(10):1729–39. doi: 10.1210/me.2014-1102.Hedrick E, et al. The nuclear orphan receptor NR4A1 regulates β1-integrin expression in pancreatic and colon cancer cells and can be targeted by NR4A1 antagonists. Mol Carcinog 2017;56(9):2066–2075.Liu X, et al. Genome-wide analysis identifies NR4A1 as a key mediator of T cell dysfunction. Nature 2019;567(7749):525–9. Epub 2019/03/01. doi: 10.1038/s41586-019-0979-8.Chen J et al. NR4A transcription factors limit CAR T cell function in solid tumours. Nature. 2019;567(7749):530–4. Epub 2019/03/01. doi: 10.1038/s41586-019-0985-x. PubMed PMID: 30814732; PMCID: PMC6546093.Seo H, et al. TOX and TOX2 transcription factors cooperate with NR4A transcription factors to impose CD8(+) T cell exhaustion. Proc Natl Acad Sci USA 2019;116(25):12410–5.Ethics ApprovalAll animal studies were carried out according to the ethical procedures approved by the Texas A&M University Institutional Animal Care and Use Committee. Approval number is 2020-0138.Abstract 732 Figure 1CDIM/NR4A1 antagonists inhibit colon tumor growth. Model for regulation of genes with GC-rich promoters by NR4A1/SP1 (A). C57BL/6 mice bearing MC-38 cells as xenografts were treated for 21 days with corn oil (control), DIM-3-Br-5-OCF3 (2.5 and 7.5 mg/kg/d), DIM-3,5-Cl2 (2.5 and 7.5 mg/kg/d) and effects on tumor volume (B), and tumor weights (C) were determined.Abstract 732 Figure 2CDIM analogs alter transcription factors expression. FACS analysis and CD4 and CD8 – specific antibodies were used to determine T-cell population in tumors of mice treated with corn oil (control), DIM-3-Br-5-OCF3 (2.5 and 7.5 mg/kg/d) and DIM-3,5-Cl2 (2.5 and 7.5 mg/kg/d) (A). Real time PCR was used to determine expression of nuclear factors (B) and cytokine (C) mRNA levels in CD8+ T-cells isolated from tumors. Results are expressed as means ± SD replicates from each treatment group and significant (pAbstract 732 Figure 3NR4A1 ligands decreases T-cell exhaustion markers. FACS analysis in tumors derived from mice treated with corn oil (control), DIM-3-Br-5-OCF3 (2.5 and 7.5 mg/kg/d) and DIM-3,5-Cl2 (2.5 and 7.5 mg/kg/d) using specific antibodies was carried out to determine percentage of CD8+ T-cells expressing T-cell exhaustion markers - PD1 (A), 2B4 (B), TIGIT (C) and TIM3 (D). Significant (p

Flavonoids are polyphenolic phytochemicals produced in fruits, nuts and vegetables and dietary consumption of these structurally diverse compounds is associated with multiple health benefits including increased lifespan, decreased cardiovascular problems and low rates of metabolic diseases. Preclinical studies with individual flavonoids demonstrate that these compounds exhibit anti-inflammatory and anticancer activities and they enhance the immune system. Their effectiveness in both chemoprevention and chemotherapy is associated with their targeting of multiple genes/pathways including nuclear receptors, the aryl hydrocarbon receptor (AhR), kinases, receptor tyrosine kinases and G protein-coupled receptors. However, despite the remarkable preclinical activities of flavonoids, their clinical applications have been limited and this is due, in part, to problems in drug delivery and poor bioavailability and these problems are being addressed. Further improvements that will expand clinical applications of flavonoids include mechanism-based precision medicine approaches which will identify critical mechanisms of action of individual flavonoids with optimal activities that can be used in combination therapies.

Background: The aryl hydrocarbon receptor (AhR) is expressed in gliomas and the highest staining is observed in glioblastomas. A recent study showed that the AhR exhibited tumor suppressor-like activity in established and patient-derived glioblastoma cells and genomic analysis showed that this was due, in part, to suppression of CXCL12, CXCR4 and MMP9. Methods: Selective AhR modulators (SAhRMs) including AhR-active pharmaceuticals were screened for their inhibition of invasion using a spheroid invasion assay in patient-derived AhR-expressing 15-037 glioblastoma cells and in AhR-silenced 15-037 cells. Invasion, migration and cell proliferation were determined using spheroid invasion, Boyden chambers and scratch assay, and XTT metabolic assays for cell growth. Changes in gene and gene product expression were determined by real-time PCR and Western blot assays, respectively. In vivo antitumorigenic activity of omeprazole was determined in SCID mice bearing subcutaneous patient-derived 15-037 cells. Results: Results of a screening assay using patient-derived 15-037 cells (wild-type and AhR knockout) identified the AhR-active proton pump inhibitor omeprazole as an inhibitor of glioblastoma cell invasion and migration only AhR-expressing cells but not in cells where the AhR was downregulated. Omeprazole also enhanced AhR-dependent repression of the pro-invasion CXCL12, CXCR4 and MMP9 genes, and interactions and effectiveness of omeprazole plus temozolomide were response-dependent. Omeprazole (100 mg/kg/injection) inhibited and delayed tumors in SCID mice bearing patient-derived 15-037 cells injected subcutaneously. Conclusion: Our results demonstrate that omeprazole enhances AhR-dependent inhibition of glioblastoma invasion and highlights a potential new avenue for development of a novel therapeutic mechanism-based approach for treating glioblastoma.

Transient abnormal myelopoiesis (TAM) also known as transient myeloproliferative disorder (TMD), a unique transient neonatal preleukaemic disorder characterized by clonal proliferation of megakaryoblasts, has been usually described to be associated with Down syndrome neonates. However, there are case reports of it occurring in neonates without Down phenotype, who are either mosaic for trisomy 21 or have trisomy 21 restricted to leukemic clone. This case report presents a case of TAM in a phenotypically normal neonate who presented in respiratory distress with features of tumour lysis syndrome (TLS) immediately after birth who was treated symptomatically and had spontaneous remission within three months.

Transforming growth factor β (TGFβ) enhances invasion of breast and lung cancer cells through phosphorylation-dependent nuclear export of the nuclear receptor 4A1 (NR4A1, Nur77). This response is inhibited by the NR4A1 antagonist 1,1-bis(3'-indoly)-1-(p-hydroxyphenyl) methane (CDIM8) and we hypothesized that similar effects would be observed in Rhabdomyosarcoma (RMS) cells. Although some kinase inhibitors block TGFβ-induced invasion of embryonal RMS (ERMS) cells, the mechanism differs from breast and lung cancer cells since NR4A1 is extranuclear in ERMS cells. However, CDIM8 blocks basal and TGFβ-induced invasion of RD and SMS-CTR ERMS cell lines but not Rh30 alveolar RMS (ARMS) cells. Moreover, this response in ERMS cells was independent of SMAD7 degradation or activation of SMAD2/SMAD3. β-Catenin silencing decreased ERMS cell invasion and CDIM8 induced proteasome-independent downregulation of β-catenin. The novel mechanism of CDIM8-mediated inhibition of basal and TGFβ-induced ERMS cell invasion was due to activation of the Bcl-2-NR4A1 complex, mitochondrial disruption, induction of the tumor suppressor-like cytokine interleukin-24 (IL-24) which in turn downregulates β-catenin expression. Thus, the NR4A1 antagonist inhibits TGFβ-induced invasion of ERMS cells through initial targeting of cytosolic NR4A1.

In 2015, massive earthquakes of 7.8 and 7.4 magnitude struck Nepal. This resulted in severe economic and infrastructural damage, not to mention many human casualties. The government of Nepal has identified 625,000 houses as fully destroyed and 180,000 houses as being partially damaged. This research is a comparative study of traditional-urban, peri-urban, and remote rural settlements of Nepal which were severely hit by the earthquake. It provides an overview of interests and perceptions of local communities in terms of the recovery process. Furthermore, this research also identifies resilience in terms of basic service recovery (basic shelter, electricity, water supply, telecommunication, groceries/food) and existing challenges in housing recovery programs. Assessing the different settlement types individually also allows for tailored policy recommendations to bridge related gaps. From the survey conducted, it can be seen that earthquake affected people’s perception of housing (re)construction has changed considerably and that they are more interested in having earthquake resistant houses after the 2015 events. Analysis also shows that, unlike in urban areas, people in rural areas tend to build stronger houses when they understand the scientific reason behind earthquake-induced damages. Lack of financing is a major hindrance for reconstruction in all study areas, and there is a need for government and financial institutes to engage to create favourable financing schemes.

In 2015, a massive earthquake of 7.8 and 7.4 magnitude struck Nepal. This resulted in severe economic and infrastructural damage, not to mention many human casualties. The government of Nepal has identified 625,000 houses as fully destroyed and 180,000 houses as being partially damaged (PDNA Vol B, Post disaster needs assessment – sector reports. Government of Nepal, Kathmandu, 2015; Sector Plans – GoN, Sector plans and financial projections -working documents. National Reconstruction Authority, Government of Nepal, Kathmandu, 2016).

Bis-indole derivatives including 1,1-bis(3’-indolyl)-1-(4-chlorophenyl)methane (DIM-C-pPhCl) and substituted quinolines such as chloroquine (CQ) and amodiaquine (AQ) are nuclear receptor 4A2 (NR4A2, Nurr1) ligands and they exhibit anti-inflammatory activities in mouse and rat models of Parkinson’s disease, respectively. However, computational modeling demonstrates that the quinoline derivatives interact with the ligand binding domain whereas the bis-indoles preferentially interact with a C-terminal cofactor binding site of NR4A2. In this study, the effects of DIM-C-pPhCl and related analogs were compared to CQ/AQ as inducers of NR4A2-responsive genes including vasoactive intestinal peptide (VIP), osteopontin (OPN), proopiomelanocortin (POMC) and neuropilin 1 (NRP1) in Panc1 and Panc28 pancreatic cancer cells. The results demonstrate that, among the bis-indole analogs, their relative potencies as inducers were structure-gene and cell context dependent. In contrast, CQ and AQ were significantly less potent than the bis-indole derivatives and, for some of the NR4A2-regulated genes, CQ and AQ were inactive as inducers. These results demonstrate that, although bis-indole and quinoline derivatives have been characterized as activators of NR4A2-dependent gene expression, these two classes of compounds exhibit different activities, indicating that they are selective NR4A2 modulators.

Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma that is primarily observed in children and adolescent, and accounts for 50% of soft tissue sarcomas in children. There are two major forms of RMS, namely embryonal RMS (ERMS) and alveolar RMS (ARMS) and these tumors are readily distinguished by their histological and genomic characteristics. ERMS patients have a better prognosis than ARMS patients and both groups undergo radiation, surgical and chemotherapeutic regiments. Many ARMS patients express different oncogenes including the histone methyltransferase G9a and the PAX3-FOXO1 fusion gene that contribute to ARMS cell growth, survival and migration/invasion. Studies in this laboratory have demonstrated that both G9a and PAX3-FOXO1 are regulated by the orphan nuclear receptor 4A1 (NR4A1) and can be targeted by bis-indole derived (CDIMs) NR4A1 antagonists. A recent study showed that the flavonoid kaempferol decreased G9a expression in gastric cancer cells and we hypothesized that kaempferol may be an NR4A1 ligand. Kamepferol and quercetin were used as models and both compounds bound NR4A1 at low μM concentrations and decreased NR4A1-dependent transactivation in ARMS cells. Functional studies showed that like CDIM/NR4A1 ligands, kaempferol and quercetin decrease ARMS cell growth, survival and migration/invasion, consistent with their activity as antagonists and inhibitors of NR4A1 regulated pro-oncogenic functions. In addition, kaempferol and quercetin also decreased expression of G9a, PAX3-FOXO1 and downstream genes regulated by them in ARMS cells further demonstrating their activity as NR4A1 antagonists. These results suggest that NR4A1-active flavonoids could be readily incorporated into chemotherapies used for treatment of ARMS patients to improve their efficacy and decrease the toxic side-effects of currently used cytotoxic drugs. Citation Format: Rupesh Shrestha, Kumaravel Mohankumar, Gregory Martin, Stephen Safe. Flavonoids are NR4A1 antagonists and target PAX3-FOXO1 and G9a in alveolar rhabdomyosarcoma cells [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 629.

The orphan nuclear receptor 4A1 (Nur77, TR3, NR4A1) is highly expressed in solid tumors and is a negative prognostic factor for survival of breast, lung, and liver cancer patients (1) and ongoing studies in our laboratory show that NR4A1 is pro-oncogenic in multiple tumor types. We have also identified a series of bis-indole derived compounds including 1,1-bis(3'-indolyl)-1-(p-hydroxyphenyl) methane (DIM-C-pPhOH; CDIM8) that act as NR4A1 antagonists that inhibit pro-oncogenic NR4A1-regulated genes and pathways in multiple cancer cell lines (2). Mechanistic studies show that many of the NR4A1-regulated genes involve interactions of NR4A1 with gene promoter DNA or with Sp-bound to GC-rich sequences and similar mechanisms have previously been observed for steroid hormone and other nuclear receptors. Our studies have identified paraspeckle component 1 (PSPC1) as an NR4A1 regulated gene in breast, lung and liver cancer cells and PSPC1 gene expression and protein levels are suppressed after treatment with C-DIM/NR4A1 antagonists. PSPC1 promotes tumorigenesis, epithelial-to-mesenchymal transition (EMT), stemness and metastasis and is a master activator of pro-metastatic switches and a potential target for anti-metastatic drugs. Knockdown of PSPC1 downregulates EMT and stemness markers in breast, lung and liver cancer cells. The novel buttressed analog of CDIM8 (DIM-C-pPhOH-3-Br-5-OCF3) at a concentration of 10 uM downregulates PSPC1 expression and its downstream targets (Snail, Slug, Nanog, OCT4, SOX2, c-myc). Knockdown of NR4A1 or SP1 inhibits PSPC1 expression in MDA-MB-231, H1299 and SNU449 cell lines respectively. These results identify PSPC1 as a novel gene that regulates EMT and invasion in these cancer cells and NR4A1 is an upstream regulator of PSPC1 gene expression. Thus, NR4A1 ligands act as an antimetastatic mimic by inhibiting PSPC1 and the mechanism of PSPC1 regulation involves ligand induced interactions of NR4A1/SP1 with a GC-rich region of the PSPC1 gene promoter. References:1.Safe S, Jin UH, Hedrick E, Reeder A, Lee SO. Minireview: Role of orphan nuclear receptors in cancer and potential as drug targets. Mol Endocrinol. 2014;28(2):157-72. 2.Mohankumar K, Li X, Sridharan S, Karki K, Safe S. Nuclear receptor 4A1 (NR4A1) antagonists induce ROS-dependent inhibition of mTOR signaling in endometrial cancer. Gynecol Oncol. 2019; 154(1):218-227. Citation Format: Kumaravel Mohankumar, Rupesh Shrestha, Stephen Safe. NR4A1 ligands inhibits PSPC1 regulated EMT and stemness markers in breast, lung and liver cancer cells [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 2428.

An integrated data platform harmonizes many disparate soil moisture data sets to better inform disaster response planners, climate scientists and meteorologists, farmers, and others.

Vegetation biomass estimates across drylands at regional scales are critical for ecological modeling, yet the low-lying and sparse plant communities characterizing these ecosystems are challenging to accurately quantify and measure their variability using spectral-based aerial and satellite remote sensing. To overcome these challenges, multi-scale data including field-measured biomass, terrestrial laser scanning (TLS) and airborne laser scanning (ALS) data, were combined in a hierarchical modeling framework. Data derived at each scale were used to validate an increasingly broader index of sagebrush (Artemisia tridentata) aboveground biomass. First, two automatic crown delineation methods were used to delineate individual shrubs across the TLS plots. Second, three models to derive shrub volumes were utilized with TLS data and regressed against destructively-sampled individual shrub biomass measurements. Third, TLS-derived biomass estimates at 5 m were used to calibrate a biomass prediction model with a linear regression of ALS-derived percent vegetation cover (adjusted R2 = 0.87, p

Fine-scale variation of vegetation structure in dryland systems, such as the Great Basin in the western US, is critical to understanding ecosystem responses to changing land-use conditions. High resolution airborne hyperspectral (HyMap) and LiDAR datasets acquired across independent collection sites can reduce uncertainty in predictive ecosystem modeling and provide a basis for regional upscaling to satellite observations of structural metrics such as cover and height. In the first part of our study, we combined ground reference and airborne data collected at three sagebrush-steppe locations and used the statistical data mining tool random forests to identify remote sensing variables most relevant to estimating shrub cover. In the second part of our study, we hypothesized that vegetation indices derived from hyperspectral satellite observations would not only reliably predict shrub cover but also be relatable to shrub height; thereby augmenting the collection of vertical structure estimates from future satellite platforms such as ICESAT-2. To test this hypothesis, we simulated HyspIRI observations to derive variables to relate to LiDAR-based estimates of shrub cover and height. We generated the same hyperspectral variables as in the first part of this study but at coarser resolution (60 m) and we again used random forests to model shrub cover and height and identify predictors of greatest importance. Overall, combining LiDAR and HyMap datasets at the airborne scale improved shrub cover model results (r2 = 0.58) compared to LiDAR alone (r2 = 0.49). Primary shrub cover variables of importance were HIQR (the interquartile range of height of all LiDAR vegetation returns), HMAD (median absolute deviation from median height of all LiDAR vegetation returns), a narrowband index sensitive to anthocyanins, the ratio of LiDAR vegetation returns to total returns, and a red to green ratio. In addition, HyspIRI-simulated narrowband vegetation indices were relatable to LiDAR-derived shrub cover and height variables (r2 ranging from 0.63 to 0.71) with relatively low root mean square error.

Leptospirosis is an important zoonotic disease in Nepal; however, there is a lack of information on sources of leptospirosis infection for people and associated risk factors. We implemented a case-control study nested within a cross-sectional survey to investigate zoonotic risks of human leptospirosis among adult, febrile patients seeking treatment in healthcare centres in Kaski District, Nepal, from April to October 2013. The study population was 239 febrile patients; the data consisted of paired blood samples; questionnaires on clinical symptoms; occupational, environmental and animal exposures; and a blood sample from animals in the household. In total, 63 cattle, 92 buffalo, 181 goats, 20 dogs and 14 rodents from 119 households were blood sampled. Serology was performed on the samples using the microscopic agglutination test (MAT) for a panel of 20 serovars with a starting dilution of 1:50. Clinical leptospirosis was defined as a titre of 1:400 or stronger, or a 4-fold or greater rise between acute and convalescent titres, or evidence of seroconversion for one or more serovars represented by a titre of

[1] Woody encroachment is a globally occurring phenomenon that contributes to the global carbon sink. The magnitude of this contribution needs to be estimated at regional and local scales to address uncertainties present in the global- and continental-scale estimates, and guide regional policy and management in balancing restoration activities, including removal of woody plants, with greenhouse gas mitigation goals. The objective of this study was to estimate carbon stored in various successional phases of woody encroachment. Using lidar measurements of individual trees, we present high-resolution estimates of aboveground carbon storage in juniper woodlands. Segmentation analysis of lidar point cloud data identified a total of 60,628 juniper tree crowns across four watersheds. Tree heights, canopy cover, and density derived from lidar were strongly correlated with field measurements of 2613 juniper stems measured in 85 plots (30 × 30 m). Aboveground total biomass of individual trees was estimated using a regression model with lidar-derived height and crown area as predictors (Adj. R2 = 0.76, p

Presentation from Ecological Society of America 2017Background/Question/MethodsRoot zone soil moisture (RZSM) is an important constraint on carbon flux, and soil moisture data may help to reduce uncertainties in carbon cycle modeling. Over the last few years, availability of RZSM data has grown considerably, due to the development of new airborne sensor capabilities (e.g. AirMOSS), wider installation of in situ networks (e.g. SoilSCAPE, COSMOS), and launch of the Soil Moisture Active-Passive (SMAP) satellite. The Oak Ridge National Laboratory Distributed Active Archive Center (ORNL DAAC; http://daac.ornl.gov) currently distributes about 0.7TB (8085 files) of RZSM data products. One key challenge that may limit effective utilization of these data is their heterogeneity in terms of (1) spatial footprints (points vs gridded representation), (2) frequency (sub-hourly to daily), (3) sub-surface measurement depths, and (4) methods of measurements (in situ vs remote-sensing).Results/ConclusionsWe have developed an integrated visualization and subsetting platform for RZSM datasets available for North America based on open-source software libraries. Harmonizing these disparate data in a single system not only adds value to the existing data but also facilitates exploratory analysis and data discovery among different groups of stakeholders. In this presentation, we describe the new RZSM data platform (available at https://airmoss.ornl.gov/visualize/ ) and share case studies that leverage the visualization system for real-world science applications.

A method is presented for the co-registration of overlapping elevation surfaces based on local slope analysis. Comparison and statistical analysis of local slope versus local elevation difference between overlapping surfaces allows for the estimation of both vertical and horizontal offsets between the two surfaces. This method is then used to re-align the flightlines of a Light Detection and Ranging (lidar) dataset collected in southern Idaho resulting in a dataset with significantly higher accuracy than the original. The relative horizontal accuracy is doubled, with a final value of approximately 25 to 30 cm, while the relative vertical accuracy is improved by several centimeters to a final value of approximately 6 to 7 cm.

With the progression of LiDAR (Light Detection and Ranging) towards a mainstream resource management tool, it has become necessary to understand how best to process and analyze the data. While most ground surface identification algorithms remain proprietary and have high purchase costs; a few are openly available, free to use, and are supported by published results. Two of the latter are the multiscale curvature classification and the Boise Center Aerospace Laboratory LiDAR (BCAL) algorithms. This study investigated the accuracy of these two algorithms (and a combination of the two) to create a digital terrain model from a raw LiDAR point cloud in a semi-arid landscape. Accuracy of each algorithm was assessed via comparison with >7,000 high precision survey points stratified across six different cover types. The overall performance of both algorithms differed by only 2%; however, within specific cover types significant differences were observed in accuracy. The results highlight the accuracy of both algorithms across a variety of vegetation types, and ultimately suggest specific scenarios where one approach may outperform the other. Each algorithm produced similar results except in the ceanothus and conifer cover types where BCAL produced lower errors.

Quantification of biophysical parameters of urban trees is important for urban planning, and for assessing carbon sequestration and ecosystem services. Airborne lidar has been used extensively in recent years to estimate biophysical parameters of trees in forested ecosystems. However, similar studies are largely lacking for individual trees in urban landscapes. Prediction models to estimate biophysical parameters such as height, crown area, diameter at breast height, and biomass for over two thousand individual trees were developed using best subsets multiple linear regression for a study area in central Oklahoma, USA using point cloud distributional metrics from an Optech ALTM 2050 lidar system. A high level of accuracy was attained for estimating individual tree height (R2 = 0.89), dbh (R2 = 0.82), crown diameter (R2 = 0.90), and biomass (R2 = 0.67) using lidar-based metrics for pooled data of all tree species. More variance was explained in species-specific estimates of biomass (R2 = 0.68 for Juniperus virginiana to 0.84 for Ulmus parviflora) than in estimates from broadleaf deciduous (R2 = 0.63) and coniferous (R2 = 0.45) taxonomic groups-or the data set analysed as a whole (R2 = 0.67). The metric crown area performed particularly well for most of the species-specific biomass equations, which suggests that tree crowns should be delineated accurately, whether manually or using automatic individual tree detection algorithms, to obtain a good estimation of biomass using lidar-based metrics. United States. National Aeronautics and Space Administration – NNS06AA57G National Institute of Food and Agriculture (U.S.) - NIFA PINEMAP Project Published version