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Application of microcapsules in medical textile has been studied and commercially developed widely in recent years. The aim of this work was to propose an eco-friendly microencapsulation by solvent evaporation method that can contribute to the manufacture of medical textile products using microcapsules. Quillaja saponin was used as bio-sourced surfactant and ethyl acetate was used as the less toxic, non-halogenated organic solvent. The influences of saponin concentration, stirring speed during emulsification step and the volume of ethyl acetate used to saturate the aqueous phase before emulsification on the microcapsule size and morphology were investigated. The results showed that eudragit microcapsules of ibuprofen with diameter in range from 17 to 34 µm, which were suitable for medical textile applications, were successfully elaborated. The saponin concentration varied from 0.025 to 0.1 wt%. The stirring speed was changed from 700 to 600 rpm. The volume of ethyl acetate used in saturation step was 0, 8 and 12 ml. In the scope of investigation, the saponin concentration and the volume of ethyl acetate used in saturation step did affect the microcapsule size and morphology while the stirring speed did not. The saturation step really helped to reduce the formation of irregular microparticles and to narrow the size distribution, but the microcapsules became more porous, weaker and were deformed significantly by drying in the fabric treatment process.

This study investigates the impact of openness on human capital in 112 countries worldwide in 2000–2019. An two-stage least square fixed-effect model with instrumental variables is used to unravel the complicated relationship between human capital and its key determinants. The empirical results show that the impacts of openness vary greatly among different groups of countries. For example, while FDI has no effect on human capital in developed and upper-middle-income developing countries, its effect is positive in lower-middle-income countries and negative in low-income countries. Exports stimulate human capital formation in developed countries but in low-income developing countries, they act as an impediment to human capital formation. Imports have a positive effect on human capital in developing countries; however, in developed countries, the effect of imports on human capital is negative. International cooperation effectively raises the level of human capital in developed, upper-middle-income and low-income countries; however, it has no effect in lower-middle-income countries. In light of mindsponge—mindspongeconomics and the SM3D knowledge management system, the study’s findings particularly shed light on the long-term development roadmap associated with human capital and openness expansion in less developed and emerging countries in years to come.

In this study, Al-Mg alloy coatings were deposited on a C45 steel substrate by using the electric arc spray technique. The corrosion behavior of the coating was studied by electrochemical measurements and salt spray tests. The electrochemical tests were performed in 3.5 wt.% NaCl solution. Polarization tests indicated that the corrosion potential of the coating is significantly more negative than that of steel substrate after 240 hours of immersion. EIS measurements showed that after 24 hours of immersion, the total impedance increased with increasing the immersion time. This could be due to sealing of pores by corrosion products which hinder further penetration of the solution through the coating. Red rust spots did not appear on the surface of the coating after 1360 hours of salt spray test. The corrosion rate of the coating decreased with the increase in salt spray test time. After the salt spray test, the coating surface pitted and the cross section of the coating appeared with many long voids. In addition, XRD analysis proved that the main corrosion product was Al(OH)3. The results obtained indicated that arc thermal sprayed Al-Mg alloy coatings can reliably protect steel structures against corrosion in chloride-containing aqueous solutions.

Primer sequences used for PCR and Sanger sequencing.

The fibroblast growth factor receptor (FGFR) signaling pathway is aberrantly activated in approximately 15% to 20% of patients with intrahepatic cholangiocarcinoma. Currently, several FGFR kinase inhibitors are being assessed in clinical trials for patients with FGFR-altered cholangiocarcinoma. Despite evidence of initial responses and disease control, virtually all patients eventually develop acquired resistance. Thus, there is a critical need for the development of innovative therapeutic strategies to overcome acquired drug resistance. Here, we present findings from a patient with FGFR2-altered metastatic cholangiocarcinoma who enrolled in a phase II clinical trial of the FGFR inhibitor, infigratinib (BGJ398). Treatment was initially effective as demonstrated by imaging and tumor marker response; however, after 8 months on trial, the patient exhibited tumor regrowth and disease progression. Targeted sequencing of tumor DNA after disease progression revealed the FGFR2 kinase domain p.E565A and p.L617M single-nucleotide variants (SNV) hypothesized to drive acquired resistance to infigratinib. The sensitivities of these FGFR2 SNVs, which were detected post-infigratinib therapy, were extended to include clinically relevant FGFR inhibitors, including AZD4547, erdafitinib (JNJ-42756493), dovitinib, ponatinib, and TAS120, and were evaluated in vitro. Through a proteomics approach, we identified upregulation of the PI3K/AKT/mTOR signaling pathway in cells harboring the FGFR2 p.E565A mutation and demonstrated that combination therapy strategies with FGFR and mTOR inhibitors may be used to overcome resistance to FGFR inhibition, specific to infigratinib. Collectively, these studies support the development of novel combination therapeutic strategies in addition to the next generation of FGFR inhibitors to overcome acquired resistance in patients.

Sequence for FGFR2-KIAA1598 fusion.

Sensitivity of the FGFR2 secondary mutations to FGFR inhibitors

Neuroimaging data analysis often requires purpose-built software, which can be challenging to install and may produce different results across computing environments. Beyond being a roadblock to neuroscientists, these issues of accessibility and portability can hamper the reproducibility of neuroimaging data analysis pipelines. Here, we introduce the Neurodesk platform, which harnesses software containers to support a comprehensive and growing suite of neuroimaging software (https://www.neurodesk.org/). Neurodesk includes a browser-accessible virtual desktop environment and a command line interface, mediating access to containerized neuroimaging software libraries on various computing platforms, including personal and high-performance computers, cloud computing and Jupyter Notebooks. This community-oriented, open-source platform enables a paradigm shift for neuroimaging data analysis, allowing for accessible, flexible, fully reproducible, and portable data analysis pipelines.

To reduce the reliance on fishmeal (FM), other protein sources have been evaluated on cultured animals. In a 60-days feeding trial, marrons (Cherax cainii) were fed a FM diet and five test diets containing 100% of plant-based protein sources such as soybean, lupin and valorised animal-based proteins such as poultry-by-product, black soldier fly and tuna hydrolysate. At the end of the trial, DNA samples from marron gut and rearing water were investigated through DNA-based 16S rRNA gene sequencing. Plant-based diets increased abundance for Aeromonas, Flavobacterium and Vogesella, whereas animal and insect proteins influenced diverse bacterial groups in the gut linked to various metabolic activities. Insect meal in the water favoured the growth of Firmicutes and lactic acid bacteria, beneficial for the marron health. Aeromonas richness in the gut and reared water signified the ubiquitous nature of the genus in the environment. The higher bacterial diversity in the gut and water with PBP and BSF was further supported by qPCR quantification of the bacterial single-copy gene, rpoB. The overall results suggested that PBP and BSF can exhibit positive and influential effects on the gut and water microbial communities, hence can be used as sustainable ingredients for the crayfish aquaculture.

Objectives: Here we describe the durable clinical outcomes of four FGFR2 fusion positive pancreatic cancer patients treated with FGFR inhibitors as well as a pancreatic specific analysis of FGFR alterations through our collaboration with Foundation Medicine, Inc. (FMI). Methods: We identified four patients with FGFR2 fusion positive metastatic pancreatic cancer through tumor RNAseq (OSU-SpARKFuse) which identifies gene fusions from cell free DNA (cfDNA). We assessed two tumor markers (Ca19-9 and FGFRDx, an FGFR-focused cell free DNA liquid biopsy assay) and various clinical outcomes (Best overall response, duration, and side effects to their treatments). Additionally, 10,146 tumor-tissue clinical cases of pancreatic cancers were assayed by hybrid-capture based comprehensive genomic profiling (Foundation Medicine, Inc.) to identify genomic alterations in >300 cancer-associated genes, MSI, and tumor mutation burden (TMB). Results: All four patients demonstrated durable clinical responses to various FGFR inhibitors. Patient 1 harbored an FGFR2-USP33 fusion and exhibited a partial response to pemigatinib for 25 months following progression on gemcitabine/abraxane and FOLFIRI chemotherapy. Patient 2 harbored an FGFR2-INA fusion with 12 months of stable disease to pazopanib after experiencing progression from chemotherapy. Patient 3 harbored an FGFR2-INA fusion and exhibited a 12+ month on-going, partial response to pemigatinib. Patient 4 harbored an FGFR2-CEP55 fusion and demonstrated stable disease on ponatinib for 15 months, a 24-month response to gemcitabine/platinum chemotherapy, and a 7+ month on-going partial response to infigratinib. Through the genomic profiling of 10,146 tumor-tissue clinical cases of pancreatic cancer, we identified 108 FGFR fusions resulting in a prevalence of 1%. We are currently investigating co-occurring and mutually-exclusive genomic alterations, including TMB and MSI-H status, for pancreatic cancer patients harboring FGFR alterations. Conclusions: This retrospective analysis provides clinical evidence that there is a subpopulation of KRAS wild-type FGFR2 fusion positive pancreatic patients who can have beneficial and lasting responses to FGFR inhibitors further suggesting a need to investigate and expand the availability of FGFR inhibitors to more cancer types. Citation Format: Leah Stein, Julie W. Reeser, Michele R. Wing, Karthikeyan Murugesan, Anoosha Paruchuri, Zachary Risch, Eric Samorodnitsky, Emily L. Hoskins, Amy Smith, Thuy Dao, Melissa Babcook, Muhammad Imam, Aharon Freud, Sameek Roychowdhury. Clinical impact of FGFR inhibitors on FGFR2 positive pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3395.

The under-representation of several ethnic groups in existing genetic databases and studies have undermined our understanding of the genetic variations and associated traits or diseases in many populations. Cost and technology limitations remain the challenges in performing large-scale genome sequencing projects in many developing countries, including Vietnam. As one of the most rapidly adopted genetic tests, non-invasive prenatal testing (NIPT) data offers an alternative untapped resource for genetic studies. Here we performed a large-scale genomic analysis of 2683 pregnant Vietnamese women using their NIPT data and identified a comprehensive set of 8,054,515 single-nucleotide polymorphisms, among which 8.2% were new to the Vietnamese population. Our study also revealed 24,487 disease-associated genetic variants and their allele frequency distribution, especially 5 pathogenic variants for prevalent genetic disorders in Vietnam. We also observed major discrepancies in the allele frequency distribution of disease-associated genetic variants between the Vietnamese and other populations, thus highlighting a need for genome-wide association studies dedicated to the Vietnamese population. The resulted database of Vietnamese genetic variants, their allele frequency distribution, and their associated diseases presents a valuable resource for future genetic studies.

The search for suitable fish meal replacements in aqua-diets is a salient agenda in the constant effort of making aquaculture practices more sustainable. In this study, we tested four customised diets composed by systematic inclusion of pre-selected fish meal substitutes, lupin kernel meal, BSF meal, TH and PBM on growth, metabolism, cytokine profile, gut morphology and microbiota of juvenile Lates calcarifer. Five isoproteic and isoenergetic diets were prepared viz. FM100 as a control (without fish meal substitute), while FM75, FM50, FM25 and FM0 indicates replacement of fish meal (FM) at 25%, 50%, 75%, and 100%, respectively by a mixture of four different pre-selected non-fish meal (NFM) ingredients. Fish fed FM100, FM75, FM50, FM25 exhibited consistent growth and haematological response, while the fish fed no fishmeal (FM0) showed significant decline in final body weight (FBW) and specific growth rate (SGR). The poor growth performance was correlated with a decrease in villous width, microvilli height and goblet cells density. A significant shift in abundance profile of Psychrobacter in the gut microbial profile of fish fed FM50 was noticed compared to fish fed FM100. The results of qRT-PCR showed up-regulated expression of innate immune responsive genes in the FM50 group. The adverse impacts on growth performance and gut health of fish fed FM0 suggest that the complete substitution of fishmeal is not advisable and the inclusion range of these alternatives should be decided for a species only after examining their effect on maximal physiological performance.

Introduction Relapsed SCLC is characterized by therapeutic resistance and high mortality rate. Despite decades of research, mechanisms responsible for therapeutic resistance have remained elusive owing to limited tissues available for molecular studies. Thus, an unmet need remains for molecular characterization of relapsed SCLC to facilitate development of effective therapies. Methods We performed whole-exome and transcriptome sequencing of metastatic tumor samples procured from research autopsies of five patients with relapsed SCLC. We implemented bioinformatics tools to infer subclonal phylogeny and identify recurrent genomic alterations. We implemented immune cell signature and single-sample gene set enrichment analyses on tumor and normal transcriptome data from autopsy and additional primary and relapsed SCLC data sets. Furthermore, we evaluated T cell-inflamed gene expression profiles in neuroendocrine (ASCL1, NEUROD1) and non-neuroendocrine (YAP1, POU2F3) SCLC subtypes. Results Exome sequencing revealed clonal heterogeneity (intertumor and intratumor) arising from branched evolution and identified resistance-associated truncal and subclonal alterations in relapsed SCLC. Transcriptome analyses further revealed a noninflamed phenotype in neuroendocrine SCLC subtypes (ASCL1, NEUROD1) associated with decreased expression of genes involved in adaptive antitumor immunity whereas non-neuroendocrine subtypes (YAP1, POU2F3) revealed a more inflamed phenotype. Conclusions Our results reveal substantial tumor heterogeneity and complex clonal evolution in relapsed SCLC. Furthermore, we report that neuroendocrine SCLC subtypes are immunologically cold, thus explaining decreased responsiveness to immune checkpoint blockade. These results suggest that the mechanisms of innate and acquired therapeutic resistances are subtype-specific in SCLC and highlight the need for continued investigation to bolster therapy selection and development for this cancer.

Background The fibroblast growth factor receptor (FGFR) signaling pathway is activated in multiple tumor types through gene amplifications, single base substitutions, or gene fusions. Multiple small molecule kinase inhibitors targeting FGFR are currently being evaluated in clinical trials for patients with FGFR chromosomal translocations. Patients with novel gene fusions involving FGFR may represent candidates for kinase inhibitors. Methods A targeted RNA-sequencing assay identified a KLK2-FGFR2 fusion gene in two patients with metastatic prostate cancer. NIH3T3 cells were transduced to express the KLK2-FGFR2 fusion. Migration assays, Western blots, and drug sensitivity assays were performed to functionally characterize the fusion. Results Expression of the KLK2-FGFR2 fusion protein in NIH3T3 cells induced a profound morphological change promoting enhanced migration and activation of downstream proteins in FGFR signaling pathways. The KLK2-FGFR2 fusion protein was determined to be highly sensitive to the selective FGFR inhibitors AZD-4547, BGJ398, JNJ-42756943, the irreversible inhibitor TAS-120, and the non-selective inhibitor Ponatinib. The KLK2-FGFR2 fusion did not exhibit sensitivity to the non-selective inhibitor Dovitinib. Conclusions Importantly, the KLK2-FGFR2 fusion represents a novel target for precision therapies and should be screened for in men with prostate cancer.

Interdigitating dendritic cell sarcoma (IDCS) is an extremely rare cancer of dendritic cell origin that lacks a standardized treatment approach. Here, we performed genomic characterization of metastatic IDCS through whole exome sequencing (WES) of tumor tissues procured from a patient who underwent research autopsy. WES was also performed on a treatment-naïve tumor biopsy sample obtained from prior surgical resection. Our analyses revealed ultra-hypermutation, defined as >100 mutations per megabase, in this patient's cancer, which was further characterized by the presence of three distinct mutational signatures including UV radiation and APOBEC signatures. To characterize clonal heterogeneity, we used the bioinformatics tool Canopy to leverage single nucleotide and copy number variants to catalog six subclones across various metastatic tumors. Truncal alterations, defined as being present in all clonal tumor cell populations, in this patient's cancer include point mutations in TP53 and CDKN2A and amplifications of c-KIT and APOBEC3A-H, which are likely driver mutations. In summary, we have performed genomic characterization evaluating tumor mutational burden (TMB) and heterogeneity in a patient with metastatic IDCS. Despite ultra-hypermutation, this patient's cancer was not responsive to treatment with PD-1 inhibition. Our results underscore the importance of characterizing clonal heterogeneity in TMB-high cancers.

Quality of produced seedlings relies mainly on the substrate on which it grows and develops. In this study, three common biomaterials – pine bark, carbonized rice husk and mushroom sawdust, were mixed with growing media in two ratios (10% and 20%) to assess their effects on growth and seedling quality of Prunus sargentii seedlings in containerized production system. Generally, 10% pine bark treatment showed the highest results among others in some parameters of seedling growth – height and root collar diameter. For total biomass, 10% rice husk treatment was highest, followed by 10% pine bark treatment. Seedling quality index at 10% rice husk treatment resulted highest (24% increase compared to control) followed by 10% pine bark treatment (20% increase compared to control). Our results suggested that both rice husk and pine bark can be used in 10% mixed ratio to growing media to gain the qualified P. sargentii seedlings in forest nurseries, which in turn, will help to establish successful reforestation in the field.

Alternative, collaborative, complimentary, integrative, natural, unconventional and holistic are the terms that are used to describe that do not conform to mainstream allopathic/western/orthodox health approaches. WHO supports traditional and alternative medicines when these have demonstrated benefits for the patient with minimal risks. In general, the harder a medical problem is to treat, the more treatment strategies exist. Patients and physicians are constantly looking for alternatives to drug therapy. Empirical and scientific evidence exists to support the benefits of acupuncture, manual therapies and several medicinal plants for chronic or mild conditions. For instance, the effectiveness of acupuncture, a popular treatment for relieving pain, has been demonstrated both through numerous clinical trials and laboratory experiments. As a result, 90% of pain clinics in the United Kingdom and 70% in Germany include acupuncture as a form of treatment. Acupressure is an easier form of Acupuncture wherein this technique of applying gentle pressure to various ‘pressure points’ on the body is used to cure various Diseases including relief of Pain. The aim of this paper is to give a introduction to acupuncture/acupressure to the general dental practitioner and to understand its history, background, current trends, the relevance and the possibilities of its practice in dentistry.

Knowledge on growth and nutrient uptake characteristics of urban trees and effective strategies to grow trees can help accomplish the goal of urban afforestation initiatives in a sustainable way. Thus, the study investigated the effects of different vermicompost (VC) application placements on the growth and nutrient uptake of three contrasting tree species (fast-growing Betula platyphylla and Larix kaempferi and slow-growing Chamaecyparis obtusa) to provide implications for growing tree stocks for sustainable urban afforestation programs. Five placement methods were used in the greenhouse trial: no fertilization (CON), surface placement (VCs), subsurface placement at 6-cm depth (VCc), bottom placement (35-cm depth (VCb)), and mixed with soil (VCm). We measured the growth parameters such as height, root collar diameter (RCD), and biomass and analyzed foliar nutrient concentrations in response to different placement treatments of VC. Relative height growth was the highest at VCc (132% (B. platyphylla), 114% (L. kaempferi)) and VCs ((57%) C. obtusa). Significant improvement in aboveground and belowground biomass growth of all species at VCs and VCc compared to the other treatments was also observed. Generally, VC treatments significantly increased N concentration compared to CON in all species. In conclusion, fertilizing the fast- and slow-growing urban tree species using VCs and/or VCc is relevant to growing high quality planting stocks for sustainable urban afforestation purposes.

Background In reclaimed land, the growth environment for plants may be unfavorable and the initial establishment and growth of seedlings could be limited because of low nutrient and water availability. Fertilization and control of understory vegetation that competes with seedlings may be of help in ameliorating soil physical and chemical properties, resulting in better seedling growth and reclamation success. However, the amount of nutrients understory vegetation absorbs in this ecological process has been rarely studied. Thus, we aimed to investigate the effect of soil amendment on biomass production and nutrient uptake by weeds in the nutrient-poor reclaimed area. We applied three levels of torrefied wood chip (TWC; 0, 2.5, and 5 Mg ha−1) and two levels of vermicompost (VC; 0 and 2.7 Mg ha−1) as soil physical improvements and organic soil amendments in reclaimed land in the Republic of Korea, with Populus euramericana used as the crop tree. Results TWC did not influence weed biomass, but 2.7 Mg ha−1 VC significantly increased weed biomass by 21% compared to 0 Mg ha−1 VC treatments. Nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) concentrations in weeds were the highest in control, but there was no statistical difference among treatments. However, VC treatment did marginally increase nutrient uptake in weeds, especially P, K, Ca, and Na. No treatments influenced crop tree height. Conclusion We conclude that VC can be used as an organic source of nutrients in reclaimed soil and that weed management is necessary to increase treatment effects on crop trees in this salt-affected reclaimed land.

Microsatellites are short, repetitive segments of DNA, which are dysregulated in mismatch repair-deficient (MMRd) tumors resulting in microsatellite instability (MSI). MSI has been identified in many human cancer types with varying incidence, and microsatellite instability-high (MSI-H) tumors often exhibit increased sensitivity to immune-enhancing therapies such as PD-1/PD-L1 inhibition. Next-generation sequencing (NGS) has permitted advancements in MSI detection, and recent computational advances have enabled characterization of tumor heterogeneity via NGS. However, the evolution and heterogeneity of microsatellite changes in MSI-positive tumors remains poorly described. We determined MSI status in 6 patients using our previously published algorithm, MANTIS, and inferred subclonal composition and phylogeny with Canopy and SuperFreq. We developed a simulated annealing-based method to characterize microsatellite length distributions in specific subclones and assessed the evolution of MSI in the context of tumor heterogeneity. We identified three to eight tumor subclones per patient, and each subclone exhibited MMRd-associated base substitution signatures. We noted that microsatellites tend to shorten over time, and that MMRd fosters heterogeneity by introducing novel mutations throughout the disease course. Some microsatellites are altered among all subclones in a patient, whereas other loci are only altered in particular subclones corresponding to subclonal phylogenetic relationships. Overall, our results indicate that MMRd is a substantial driver of heterogeneity, leading to both MSI and subclonal divergence. Implications: We leveraged subclonal inference to assess clonal evolution based on somatic mutations and microsatellites, which provides insight into MMRd as a dynamic mutagenic process in MSI-H malignancies.

Cholangiocarcinoma is a highly aggressive and lethal malignancy, with limited treatment options available. Recently, FGFR inhibitors have been developed and utilized in FGFR-mutant cholangiocarcinoma; however, resistance often develops and the genomic determinants of resistance are not fully characterized. We completed whole-exome sequencing (WES) of 11 unique tumor samples obtained from a rapid research autopsy on a patient with FGFR-fusion-positive cholangiocarcinoma who initially responded to the pan-FGFR inhibitor, INCB054828. In vitro studies were carried out to characterize the novel FGFR alteration and secondary FGFR2 mutation identified. Multisite WES and analysis of tumor heterogeneity through subclonal inference identified four genetically distinct cancer cell populations, two of which were only observed after treatment. Additionally, WES revealed an FGFR2 N549H mutation hypothesized to confer resistance to the FGFR inhibitor INCB054828 in a single tumor sample. This hypothesis was corroborated with in vitro cell-based studies in which cells expressing FGFR2–CLIP1 fusion were sensitive to INCB054828 (IC50 value of 10.16 nM), whereas cells with the addition of the N549H mutation were resistant to INCB054828 (IC50 value of 1527.57 nM). Furthermore, the FGFR2 N549H secondary mutation displayed cross-resistance to other selective FGFR inhibitors, but remained sensitive to the nonselective inhibitor, ponatinib. Rapid research autopsy has the potential to provide unprecedented insights into the clonal evolution of cancer throughout the course of the disease. In this study, we demonstrate the emergence of a drug resistance mutation and characterize the evolution of tumor subclones within a cholangiocarcinoma disease course.

Background: Cholangiocarcinoma is an aggressive and deadly bile duct cancer. Due to its poor prognosis and limited treatments beyond first-line therapy, in-depth characterization of cholangiocarcinoma is needed to facilitate the development of novel therapies. Tumor heterogeneity has emerged as a contributor to treatment resistance, but it is largely unknown how it contributes to cholangiocarcinoma. Characterization of tumor heterogeneity in cholangiocarcinoma has largely been limited to biopsies and surgical resections and may not accurately reflect the complex and heterogeneous nature of the disease. A partnership with patients and their families for body donation is a powerful strategy to characterize the landscape of tumor heterogeneity in cholangiocarcinoma. Study Design: We performed whole exome sequencing (WES) of a pre-treatment tumor biopsy and multiple tumor samples collected through rapid research autopsy from 9 patients with metastatic cholangiocarcinoma. This will allow us to determine spatial heterogeneity, delineate cancer phylogeny, and identify spatial origin of treatment-resistant clones as well as drivers of metastasis and recurrence. Results and Conclusions: We completed research autopsies of 9 patients with metastatic cholangiocarcinoma including 5 with FGFR alterations, 1 with an IDH1 mutation, 1 with a BRCA mutation, 1 with a BAP1 splice site mutation, and 1 with hypermutation by an unknown mechanism. We observed varying patterns of clonal evolution in this cohort including persistent, sensitive, and emergent clones. One patient with metastatic cholangiocarcinoma with a novel FGFR2-CLIP1fusion received the FGFR inhibitor, INCB054828 and had a 5.5-month response but unfortunately progressed and succumbed to his disease. Multi-site WES and analysis of tumor heterogeneity through subclonal inference identified four genetically distinct clonal cancer cell populations, two of which were only observed in post-treatment samples (emergent clones). Additionally, WES revealed an FGFR2N549H mutation in a single tumor sample hypothesized to confer resistance to INCB054828. This hypothesis was corroborated in vitro, in which cells expressing the FGFR2-CLIP1 fusion protein were sensitive to INCB054828 (IC50of 10.16 nM), whereas cells expressing FGFR2-CLIP1 N549H were resistant (IC50of 1257.57 nM). Furthermore,the FGFR2 N549H mutation displayed cross-resistance to other selective FGFR inhibitors, but remained sensitive to the non-selective inhibitor, ponatinib. Cancer Relevance: These studies will augment our understanding of tumor heterogeneity in cholangiocarcinoma. The broader impact of this research includes opportunities to develop new therapies for cholangiocarcinoma, centralized storage of cholangiocarcinoma tumor samples, and demonstration of how research autopsy can be applied to study tumor heterogeneity in other cancer types. Citation Format: Melanie A. Krook, Russell Bonneville, Hui-Zi Chen, Julie W. Reeser, Michele R. Wing, Jharna Miya, Amy M. Smith, Anoosha Paruchuri, Thuy Dao, Dorrelyn M. Martin, Eric Samorodnitsky, Kaitlin R. Baker, Cynthia Timmers, Kristin Dittmar, Sharon Cole, Lianbo Yu, Aharon G. Freud, Patricia Allenby, Sameek Roychowdhury. Rapid autopsy of metastatic cholangiocarcinoma demonstrates diverse patterns of clonal heterogeneity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2909.

Genomic characterization of recurrent small cell lung cancer (SCLC) is needed to dissect the molecular mechanisms of relapse, metastatic dissemination, and chemoresistance. SCLC is a highly aggressive, lethal neuroendocrine cancer arising from the bronchial epithelium that develops in heavy smokers. Although many SCLC patients demonstrate sensitivity to first-line platinum-doublet chemotherapy, the response is short-lived and nearly all patients progress during therapy or relapse within several months of completing treatment. Treatment options beyond first-line chemotherapy are limited and ineffective. Metastatic SCLC has a poor prognosis and is associated with a 5-year overall survival rate of 5-10%. A well-characterized genetic hallmark of SCLC is the concurrent inactivation of tumor suppressor genes TP53 and RB1. Additional SCLC candidate driver genes have been reported and include CREBBP, MLL, NOTCH1-4, SOX2 and MYC. However, most SCLC tumors profiled in these studies were obtained from treatment-naïve patients. Therefore, genomic alterations that are acquired during therapy and drive the development of chemoresistance in recurrent SCLC remain undefined. We present results from whole exome sequencing (WES) of multiple metastatic tumors procured from five treatment-refractory SCLC patients who underwent research autopsy. WES was also performed on pre-treatment samples from three of these patients. Our results showed universal TP53 alterations and to a lesser extent RB1 mutations in our SCLC cohort. Next, we utilized bioinformatics methods to analyze clonal heterogeneity and evolution in these SCLC patients. This analysis demonstrated the existence of multiple clones of tumor cells in both the pre-treatment and treatment-resistant autopsy SCLC samples, suggesting that tumor heterogeneity occurs early in SCLC development and is subsequently maintained. Furthermore, allele-specific analysis of copy number variations in our SCLC cohort demonstrated near universal deletion of a region on chromosome 5q containing the tumor suppressor APC, consistent with a recently reported role for deregulated WNT signaling in chemoresistance development in relapsed SCLC. Finally, in one SCLC patient, we identified PTEN deletion (loss of heterozygosity) as a potential mechanism of metastasis specifically to the brain. In conclusion, we have performed genomic characterization of recurrent SCLC, defined by significant clonal diversity, through research autopsy and identified candidate genetic drivers of treatment resistance and organ-specific metastasis. Citation Format: Hui-Zi Chen, Russell Bonneville, Melanie A. Krook, Michele R. Wing, Julie W. Reeser, Jharna Miya, Anoosha Paruchuri, Eric Samorodnitsky, Lianbo Yu, Amy M. Smith, Thuy Dao, Dorrelyn Martin, Qishan Guo, Hailey Magenheim, Aharon G. Freud, Sharon Cole, Gregory Otterson, Peter Shields, David P. Carbone, Patricia Allenby, Sameek Roychowdhury. Genomic characterization of recurrent small cell lung cancer through research autopsy reveals clonal diversity and candidate driver of chemoresistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 748.

Background. The fibroblast growth factor receptor (FGFR) signaling pathway is aberrantly activated in approximately 20% of cases of cholangiocarcinoma through various genomic alterations including gene fusions, point mutations, and copy number amplifications. Currently, several FGFR kinase inhibitors are being assessed in clinical trials for patients with FGFR-altered cholangiocarcinoma. Despite evidence of initial responses and disease control, virtually all tumors eventually develop drug resistance. Overcoming acquired drug resistance to FGFR inhibitors has not been comprehensively studied. Methods. Apatient with metastatic cholangiocarcinoma and an activating FGFR2-KIAA1598fusion was enrolled on a Phase 2 clinical trial for the oral FGFR inhibitor, BGJ398. BGJ398 treatment was initially effective with radiographic and tumor marker response; however, after eight months of BGJ398 therapy, the patient developed disease progression. Sequencing of a post-progression liver biopsy revealed a novel secondary mutation (E566A) in the FGFR2 kinase domain, which was absent in the pre-treatment biopsy suggesting that this mutation may be responsible for the resistance to BGJ398. To confirm this, we conducted in vitrostudies to characterize the sensitivity of the FGFR2 secondary mutation to FGFR inhibitors. Results and Conclusions. In vitrocharacterization of the fusion and putative resistance mutation demonstrated that FGFR2-KIAA1598 WT cells were sensitive to BGJ398 with (IC50value of 10.95 nM) whereas the FGFR2-KIAA1598 E566A cells were resistant (IC50values of 422.4 nM). We investigated additional selective (AZD4547 and JNJ42756493), non-selective (dovtinib and ponatinib), and irreversible (TAS120) FGFR inhibitors. FGFR2-KIAA1598 WT cells were sensitive to AZD4547, JNJ42756493, and TAS120, whereas FGFR2-KIAA1598 E566A cells were resistant to these inhibitors. Interestingly, neither the FGFR2-KIAA1598 WT cells nor the FGFR2-KIAA1598 E566A cells were sensitive to dovitinib, a nonspecific tyrosine kinase inhibitor. Lastly, both populations of cells were equally sensitive to ponatinib, a non-selective FGFR kinase inhibitor. Proteomic analysis by reverse phase protein array (RPPA) revealed upregulation of the mTOR/PI3K/AKT pathway in fusion cells compared to empty controls (FDR: 0.00618), and this expression was further potentiated in FGFR2-KIAA1598 E566A (FDR: 1.96e-19). Furthermore, combination studies with the above FGFR inhibitors and the mTOR inhibitor, INK128, yielded synergistic antitumor effects suggesting that combination therapy may be able to overcome resistance to FGFR inhibitors. Significance. In conclusion, further elucidation of the mechanisms driving resistance to FGFR inhibitors will support the development of next generation FGFR inhibitors as well as combination therapeutic strategies. Citation Format: Melanie A. Krook, Alexandria M. Lenyo, Maxwell J. Wilberding, Hannah D. Barker, Mikayla Dantuono, Hui-Zi Chen, Julie W. Reeser, Michele R. Wing, Jharna Miya, Eric Samorodnitsky, Amy M. Smith, Thuy Dao, Dorrelyn M. Martin, Kristin Dittmar, Kristen K. Ciombor, John L. Hays, Aharon G. Freud, Sameek Roychowdhury. Efficacy of FGFR inhibitors and combination therapies for acquired resistance in FGFR2-fusion cholangiocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 335.

We investigated resistance switching behavior of the Ag/ZnO/Ti structures for random access memory devices. These films were prepared on glass substrate by dc sputtering technique at room temperature. The resistance switching follows unipolar switching mode with small switching voltages (0.4 V – 0.6 V). Our results figured out that the Ag/ZnO/Ti/Glass structure is a candidate structure for nonvolatile data storage applications.

This paper presents the synthesis and characterization of Y3Fe5O12, Gd3Fe5O12 and Dy3Fe5O12 nanoparticles by sol-gel method using initial salts solution of Y(NO3)3, Gd(NO3)3, Dy(NO3)3, Fe(NO3). The lattice constant, crystallite size and particle morphology of these ferrite garnet nanoparticles were studied by using X-ray diffraction and transmission electron microscopy (TEM). The results showed that the garnet samples prepared by this method were formed at 800oC, which is lower than the sintering temperature for ceramic bulk samples (1400oC). The particle size is in the range 25- 40 nm as observed via TEM image and the average particle size was found to be 37nm using Debye- Scherrer formula.