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Objectives SSc is a devastating autoimmune disease characterized by fibrosis and obliterative vasculopathy affecting the skin and visceral organs. While the processes mediating excessive extracellular matrix deposition and fibroblast proliferation are clear, the exact link between autoimmunity and fibrosis remains elusive. Th17 cells have been proposed as critical drivers of profibrotic inflammation during SSc, but little is known about the immune components supporting their pathogenic role. Our aim was to determine cytokine responses of stimulated monocyte-derived dendritic cells (Mo-DCs) and to determine how they influence T-cell cytokine production in SSc. Material and methods Dendritic cells (DCs) activate and shape T cell differentiation by producing polarizing cytokines. Hence, we investigated the cytokine responses of monocyte-derived DCs (Mo-DCs) from patients with limited cutaneous SSc (lcSSc), diffuse cutaneous SSc (dcSSc) and healthy controls (HCs) after stimulation with toll-like receptor (TLR) agonists. Also, using co-culture assays, we analysed T cell subpopulations after contact with autologous TLR-activated Mo-DCs. Results In general, we observed an increased production of Th17-related cytokines like IL-1β, IL-17F, IL-21 and IL-22 by SSc compared with HC Mo-DCs, with variations between lcSSc vs dcSSc and early- vs late-stage subgroups. Noticeably, we found a significant increment in IL-33 production by Mo-DCs in all SSc cases regardless of their clinical phenotype. Strikingly, T cells displayed Th2, Th17 and dual Th2–Th17 phenotypes after exposure to autologous TLR-stimulated Mo-DCs from SSc patients but not HCs. These changes were pronounced in individuals with early-stage dcSSc and less significant in the late-stage lcSSc subgroup. Conclusions Our findings suggest that functional alterations of DCs promote immune mechanisms favouring the aberrant T cell polarization and profibrotic inflammation behind clinical SSc heterogeneity.

Strengthening infection prevention and control programs in nonacute care settings is a national priority. Efforts require thorough and ongoing appraisal of organizational structures, human resources including personnel training and competencies, system challenges and adaptive strategies implemented. Assessment of those in infection preventionist (IP) roles outside of the acute care setting is necessary to capture ongoing changes and challenges in the IP profession.This cross-sectional study utilized data derived from the 2020 APIC MegaSurvey and applied descriptive and bivariate analyses to describe the state of infection prevention and control programs and personnel across nonacute clinical settings in the United States.Of 1,991 respondents, 57% of frontline IPs or administration/director IPs (1,051) indicated working in 1 or more nonacute care clinical settings. Of these, 33% (343) worked exclusively in only 1 type of nonacute care setting. Consistent with findings from the 2015 APIC MegaSurvey, IPs employed in nonacute care settings are a homogenous group with 88% of respondents indicating they are white, non-Hispanic (88%), female (94%), with nursing as their primary discipline (95%). A notable change in the proportion of time spent on health care-associated infection (HAI) activities in general was found, with a 31% decrease in reported time spent compared to respondents from the 2015 survey. Nearly half (47%) of respondents reported an annual salary of $50,000-$80,000; only 35% of respondents reported they were satisfied with their overall compensation. More than half (57%) of respondents reported having 5 or less years' experience in IPC and the majority, 82% reported they expected to be working in the IP profession in the next 5 years.The majority of IPs in nonacute care settings also worked in acute care. Of those who exclusively worked in nonacute care settings, they were predominately female, white, and had an educational background in nursing. A decrease in time spent on HAI activities was noted compared to respondents in 2015. Although the 2020 APIC MegaSurvey captured information previously not assessed in 2015, further studies are necessary to more robustly characterize the IP profession in nonacute care settings. Enhancements to current resources and services provided by APIC may serve to fill gaps in nonacute care settings related to gaining experience in research, general expertise, advocacy, and diversity.

This article highlights the gap in formal credit-based community college student leadership development opportunities. The authors offer eight leadership competencies to guide the development of a stackable community college leadership development certificate. This certificate serves to enhance equitable workplace development skills for the large demographic of racially minoritized students who access postsecondary education through the community college system.

Preventing COVID-19 infections and ensuring resident safety in the nursing home (NH) setting requires collaboration among NH leadership, staff, residents, and their advocates including families, caregivers, and nursing home ombudsmen. To address this multi-collaborative need, the University of North Texas Health Science Center Geriatric Workforce Enhancement Program developed training in two focus areas. The first focused on resilience training for Certified Nurse Aides (CNAs) with the United Way that addresses factors influencing well-being, economic stability, mental health, and other social determinants. The second focused on training residents, families, ombudsmen, and other resident advocates to become active partners with NH leadership and staff in the prevention and control of healthcare acquired infections. To ensure training reflects the needs and preferences of key NH stakeholders, we developed a Nursing Home Advisory Council in partnership with state Ombudsman Programs in Texas, Louisiana, and New Mexico.

In this article for Journal Club commentary entitled "Racial Disparities in Catheter Related Urinary Tract Infections Among Elderly Trauma Patients in the US", Keneally et al, conducted a study with the goal of assessing the role of social disparities in catheter associated urinary tract infections (CAUTIs). This cross-sectional study utilized secondary data to determine the possible CAUTI risk in ventilated, older (≥ 65 years of age) trauma patients exploring possible racial structural bias in healthcare. The analysis addressed the following questions in this specific population:While this study does consider race and discusses structural biases, which are important and sparsely researched in healthcare-associated infection (HAI) outcomes, the practice implementations are somewhat limited due to study design and analysis. Therefore, the focus of this Journal Club commentary will be reviewing basic steps Infection Preventionists (IPs) can take to critically appraise the literature for application in their facility's patient population.

Previous studies indicate variability in the accurate application of National Healthcare Safety Network surveillance criteria with limited data on possible contributing factors. In this cross-sectional, convenience sampled web-based survey sent to members of Texas infection prevention and control organizations, training, experience, and time spent on surveillance was collected and assessed including 2 case studies. Our results indicate correct identification of catheter-associated urinary tract infection (CAUTI) and central line-associated bloodstream infection (CLABSI) criteria may be associated with 2019 National Healthcare Safety Network training (CAUTI: aOR = 0.17, 95% CI: 0.04, 0.80; CLABSI: aOR = 0.45, 95% CI: 0.045, 4.56) and increased years of infection prevention experience (CAUTI: aOR = 1.35, 95% CI: 0.42, 4.33; CLABSI: aOR = 1.23, 95% CI: 0.24, 6.38). Routinely performing more hours of surveillance may increase accuracy of CLABSI identification, but not CAUTI.

En la actualidad, la influencia ejercida por la sociedad hacia la actividad empresarial, seguida de la importancia de ser responsable y sustentable ha permitido resaltar el efecto y relación del emprendimiento con los ámbitos económico, ambiental y social. El sector de bebidas alcohólicas ha incrementado su participación en el mercado mexicano, en la que existen un pequeño grupo de empresas que producen y comercializan destilados artesanales que se caracterizan por requerir desde el inicio de operaciones diferentes acciones para poder competir con las grandes empresas comerciales. Para ubicar el papel en el que las empresas se encuentran, se desarrolló metodológicamente una investigación cualitativa, de carácter exploratorio y descriptivo, de la que se pudo determinar el grado de relación entre el contexto de las actividad empresarial en el sector de bebidas artesanales y la relevancia de integrar de la sustentabilidad en su actividad empresarial, cuyo análisis se llevó a cabo mediante la aplicación de entrevistas con el objetivo de verificar los elementos que intervienen al momento de establecer actividades que involucren el aspecto social, ambiental y económico de la empresa en al menos 40 empresas cuyo giro son cooperativas, aquellas de reciente creación describieron no pueden establecer desde la administración básica pilares sólidos debido a la falta de un especialista en administración, lo que provoca no poder avanzar a una administración sustentable, y así dar respuesta a las demandas de la problemática actual.

Arboviruses such as Chikungunya (CHIKV), Dengue (DENV), and Zika virus (ZIKV) have emerged as a significant public health concern in Mexico. The existing literature lacks evidence regarding the dispersion of arboviruses, thereby limiting public health policy&rsquo, s ability to integrate the diagnosis, management, and prevention. This study seeks to reveal the clinical symptoms of CHIK, DENV, and ZIKV by age group, region, sex, and time across Mexico. The confirmed cases of CHIKV, DENV, and ZIKV were compiled from January 2012 to March 2020. Demographic characteristics analyzed significant clinical symptoms of confirmed cases. Multinomial logistic regression was used to assess the association between clinical symptoms and geographical regions. Females and individuals aged 15 and older had higher rates of reported significant symptoms across all three arboviruses. DENV showed a temporal variation of symptoms by regions 3 and 5, whereas ZIKV presented temporal variables in regions 2 and 4. This study revealed unique and overlapping symptoms between CHIKV, DENV, and ZIKV. However, the differentiation of CHIKV, DENV, and ZIKV is difficult, and diagnostic facilities are not available in rural areas. There is a need for adequately trained healthcare staff alongside well-equipped lab facilities, including hematological tests and imaging facilities.

Background Previous studies indicate inconsistent knowledge and application of the National Healthcare Safety Network (NHSN) surveillance definitions used to report Healthcare Associated Infections (HAI). Our study aims to determine factors associated with accurate application of the surveillance criteria. Methods Members of Texas Infection Prevention (IP) professional organizations were emailed a web-based survey in January 2020. Respondents answered demographic and other questions related to 2019 NHSN trainings completed and applied NHSN definitions to case scenarios on Catheter-Associated Urinary Tract Infection (CAUTI) and Central Line-Associated Bloodstream Infection (CLABSI). This cross-sectional study used logistic regression models to estimate associations between years of experience, hours spent on surveillance, NHSN trainings, and respondents’ ability to identify correct HAI criterion. Results Eighty-eight IPs responded to the survey. Twenty-five of 65 respondents (38%) who completed CAUTI training identified correct CAUTI criterion and 6 of 63 (10%) who completed CLABSI training identified correct CLABSI criterion. CAUTI training was associated with higher odds of accurately identifying CAUTI (Odds Ratio (OR) = 4.92, 95% Confidence Interval (CI): 1.2, 20.5) after adjusting for years of experience and hours spent on surveillance. Similar patterns of association were observed between completion of CLABSI training and correctly identifying CLABSI, but the result was not significant (OR = 2.35, 95% CI: 0.2, 23.8). Compared to those spending less time on NHSN surveillance ( 16 hours/week) had lower odds of identifying CAUTI but higher odds of identifying CLABSI. Conclusions Limited studies demonstrate variability in the accurate application of NHSN surveillance definitions. Possible factors contributing to this variability are also lacking. Our results suggest that recent NHSN training and more years of IP experience may be associated with increased accuracy of HAI identification. However, spending more time with NHSN surveillance every week may not improve one's ability to identify CAUTI.

Neuroblastoma is the most commonly diagnosed extracranial tumor in the first year of life. Approximately 9% of neuroblastoma patients present germline or somatic aberrations in the gene encoding for anaplastic lymphoma kinase (ALK). This increases in high-risk neuroblastomas, which have a 14% frequency of ALK aberrations at the time of diagnosis and show increasing numbers at relapse. Abrogating ALK activity with kinase inhibitors is employed as clinical therapy in malignancies such as non-small cell lung cancer and has shown good results in pediatric inflammatory myofibroblastic tumors and anaplastic large cell lymphomas. A phase I clinical trial of the first generation ALK inhibitor, crizotinib, in neuroblastoma patients showed modest results and suggested that further investigation was needed. Continuous development of ALK inhibitors has resulted in the third generation inhibitor repotrectinib (TPX-0005), which targets the active kinase conformations of ALK, ROS1 and TRK receptors. In the present study we investigated the effects of repotrectinib in a neuroblastoma setting in vitro and in vivo. Neuroblastoma cell lines were treated with repotrectinib to investigate inhibition of ALK and to determine its effect on proliferation. PC12 cells transfected with different ALK mutant variants were used to study the efficacy of repotrectinib to block ALK activation/signaling. The in vivo effect of repotrectinib was also analyzed in a neuroblastoma xenograft model. Our results show that repotrectinib is capable of inhibiting signaling activity of a range of ALK mutant variants found in neuroblastoma patients and importantly it exhibits strong antitumor effects in a xenograft model of neuroblastoma.

Over the last decade Anaplastic Lymphoma Kinase (ALK), a receptor tyrosine kinase (RTK), has been identified as a fusion partner in a diverse variety of translocation events resulting in oncogenic signaling in many different cancer types. In tumors where full-length ALK RTK itself is mutated, such as neuroblastoma, the picture regarding the role of ALK as an oncogenic driver is less clear. Neuroblastoma is a complex and heterogeneous tumor that arises from the neural crest-derived peripheral nervous system. Although high-risk neuroblastoma is rare, it often relapses and becomes refractory to treatment. Thus, neuroblastoma accounts for 10-15% of all childhood cancer deaths. Since most cases are in children under the age of two, understanding the role and regulation of ALK during neural crest development is an important goal in addressing neuroblastoma tumorigenesis. An impressive array of tyrosine kinase inhibitors (TKIs) that act to inhibit ALK have been FDA approved for using in ALK-driven cancers. ALK TKIs bind differently within the ATP-binding pocket of the ALK kinase domain and have been associated with different resistance mutations within ALK itself that arise in response to therapeutic use, particularly in ALK fusion-positive NSCLC. This patient population has highlighted the importance of considering the relevant ALK TKI to be used for a given ALK mutant variant. In this poster we discuss ALK in neuroblastoma, as well as the use of ALK TKIs and other strategies to inhibit tumor growth. Current efforts combining novel approaches and increasing our understanding of the oncogenic role of ALK in neuroblastoma are aimed at improving efficacy of ALK TKIs as precision medicine options in the clinic. Citation Format: Jikui Guan, Diana Cervantes-Madrid, Joachim Siaw, Wasi Alam, Ganesh Umapathy, Marcus Borenäs, Dan Lind, Joanna Szydzik, Kathrin Pfeifer, Patricia Mendoza-Garcia, Ruth Palmer, Bengt Hallberg. Targeting anaplastic lymphoma kinase in neuroblastoma [abstract]. In: Proceedings of the AACR Special Conference on the Advances in Pediatric Cancer Research; 2019 Sep 17-20; Montreal, QC, Canada. Philadelphia (PA): AACR; Cancer Res 2020;80(14 Suppl):Abstract nr A36.

There is a strong rationale for targeting the metabolic alterations of cancer cells. The most studied of these are the higher rates of glycolysis, glutaminolysis and de novo synthesis of fatty acids (FAs). Despite the availability of pharmacological inhibitors of these pathways, no preclinical studies targeting them simultaneously have been performed. In the present study it was determined whether three key enzymes for glycolysis, glutaminolysis and de novo synthesis of FAs, hexokinase-2, glutaminase and fatty acid synthase, respectively, were overexpressed as compared to primary fibroblasts. In addition, we showed that at clinically relevant concentrations lonidamine, 6-diazo-5-oxo-L-norleucine and orlistat, known inhibitors of the mentioned enzymes, exerted a cell viability inhibitory effect. Genetic downregulation of the three enzymes also reduced cell viability. The three drugs were highly synergistic when administered as a triple combination. Of note, the cytotoxicity of the triple combination was low in primary fibroblasts and was well tolerated when administered into healthy BALB/c mice. The results suggest the feasibility and potential clinical utility of the triple metabolic targeting which merits to be further studied by using either repositioned old drugs or newer, more selective inhibitors.

Anaplastic lymphoma kinase (ALK) is a tyrosine kinase receptor that is a clinical target of major interest in cancer. Mutations and rearrangements in ALK trigger the activation of the encoded receptor and its downstream signaling pathways. ALK mutations have been identified in both familial and sporadic neuroblastoma cases as well as in 30 to 40% of relapses, which makes ALK a bona fide target in neuroblastoma therapy. Tyrosine kinase inhibitors (TKIs) that target ALK are currently in clinical use for the treatment of patients with ALK-positive non-small cell lung cancer. However, monotherapy with the ALK inhibitor crizotinib has been less encouraging in neuroblastoma patients with ALK alterations, raising the question of whether combinatorial therapy would be more effective. In this study, we established both phosphoproteomic and gene expression profiles of ALK activity in neuroblastoma cells exposed to first- and third-generation ALK TKIs, to identify the underlying molecular mechanisms and identify relevant biomarkers, signaling networks, and new therapeutic targets. This analysis has unveiled various important leads for novel combinatorial treatment strategies for patients with neuroblastoma and an increased understanding of ALK signaling involved in this disease.

Activation of the RAS-RAF-MEK-ERK signaling pathway is implicated in driving the initiation and progression of multiple cancers. Several inhibitors targeting the RAS-MAPK pathway are clinically approved as single- or polyagent therapies for patients with specific types of cancer. One example is the MEK inhibitor trametinib, which is included as a rational polytherapy strategy for treating EML4-ALK-positive, EGFR-activated, or KRAS-mutant lung cancers and neuroblastomas that also contain activating mutations in the RAS-MAPK pathway. In addition, in neuroblastoma, a heterogeneous disease, relapse cases display an increased rate of mutations in

Background Although evolution is the driving force behind many of today’s major public health and agriculture issues, both journalists and scientific researchers often do not use the term “evolve” in discussions of these topics. Methods In a total of 1,066 articles and 716 papers selected from 25 US newspapers and 34 scientific journals, we assess usage of the word “evolve” and its substitute words in the contexts of cancer tumor drug resistance, HIV drug resistance, mosquito insecticide resistance, and weed pesticide resistance. Results We find significant differences in the use of “evolve” among fields and sources. “Evolve” is used most when discussing weed pesticide resistance (25.9% in newspapers, 52.4% in journals) and least when discussing cancer tumor drug resistance (3.9% in newspapers, 9.8% in journals). On average, scientific journals use “evolve” more often (22.2%) than newspapers (7.8%). Different types of journals (general science, general clinical, cancer specific, and drug resistance specific) show significantly different “evolve” usages when discussing cancer tumor drug resistance. Discussion We examine potential explanations of these findings, such as the relatively recent framing of cancer in evolutionary terms, before looking at consequences of low “evolve” usage and of differential “evolve” usage across fields. Use of the word “evolve” may not reflect current understanding of the problems we examine. However, given that our ability to tackle resistance issues relies upon accurate understandings of what causes and exacerbates resistance, use of the word “evolve” when called for may help us confront these issues in the future.

Electrical power systems must balance demand and generation almost instantly to maintain frequency. This task is performed at each power station with the load frequency controller. On wider areas, the energy management systems supervise generation control automatically to preserve stability and reliability. Due to the unpredictability of the demand, the control response must be as fast as possible to prevent larger deviations that could lead to cascading failures and outages. In order to understand the dynamic behavior and capabilities of the controllers, real time simulation offers a great advantage over its offline predecessors, not only for operational training but also for prototyping. This paper studies the effectiveness of the real time simulator in the analysis of a power system load frequency control.

The aim of the present study was to investigate in vivo the feasibility and efficacy of the combination of lonidamine (LND), 6-diazo-5-oxo-L-norleucine (DON) and orlistat to simultaneously target glycolysis, glutaminolysis and de novo synthesis of fatty acids, respectively. The doses of LND and DON used in humans were translated to mouse doses (77.7 mg/kg and 145.5 mg/kg, respectively) and orlistat was used at 240 mg/kg. Three schedules of LND, DON and orlistat at different doses were administered by intraperitoneal injection to BALB/c mice in a 21-day cycle (schedule 1: LND, 0.5 mg/day; DON, 0.25 mg/day 1, 5 and 9; orlistat, 240 mg/kg/day; schedule 2: LND, 0.1 mg/day; DON, 0.5 mg/day 1, 5 and 9; orlistat, 240 mg/kg/day; schedule 3: LND, 0.5 mg/day; DON, 0.08 mg/day 1, 5 and 9; orlistat, 360 mg/kg/day) to assess tolerability. To determine the antitumor efficacy, a syngeneic tumor model in BALB/c mice was created using colon cancer CT26.WT cells, and a xenogeneic tumor model was created in nude mice using the human colon cancer SW480 cell line. Mice were treated with schedule 1. Animals were weighed, clinically inspected during the experiment and the tumor volume was measured at day 21. The 3 schedules assessed in the tolerability experiments were well tolerated, as mice maintained their weight and no evident clinical signs of toxicity were observed. Combination treatment with schedule 1 significantly decreased tumor growth in each mouse model. No evident signs of toxicity were observed and mice maintained their weight during treatment. The triple metabolic blockade of the malignant phenotype appears feasible and promising for cancer therapy.

This paper attempts to dissolve the issue of prostitution, which is still framed as a radical feminist issue pertaining to gender inequality and the vulnerability women face in such a career; rather, the idea post feminists (pro-prostitution feminists) hold believe prostitution, and more specifically, high class (call girl) prostitution is an empowering, positive and liberating occupation that women should be freely allowed to enter without stigmatization. We look at numerous women who have provided positive accounts of their lives as high end prostitutes, even when beholding university educations and Ph.Ds. After extensive secondary data collection to support these women’s experiences, we find high end prostitution can in fact empower, build confidence in, and gratify women.

There is a shortage of therapeutical agents for invasive cervical cancer in late stages of development; however, a number of promising molecules are currently in early phases of development.This review briefly discusses the current achievements in treating cervical cancer with an emphasis in emerging agents based on a literature search on pubmed and related sites for cervical cancer information. This is not a systematic review.In advanced disease, modest survival gains have been achieved with cisplatin doublets. Contrariwise, chemoradiation has increased survival rates in locally advanced disease, but there is still room for improvement. Anti-vascular endothelial growth factor and anti-epidermal growth factor receptor monoclonal antibodies are promising molecules that are at present in late-phase development, but a high number of miscellaneous agents are in early development. Strong experimental bases support that the 'Achilles' heel' of cervical cancer are the HPV-E6/E7 oncogenes. Unfortunately, agents aimed at targeting these cervical cancer-driven players are found in very early development; hence, major research efforts must be focused on developing technological strategies for their effective targeting using nucleic acid-based vehicles for safe and effective delivery to cancer cells as well as accelerating the search for small-molecule inhibitors of E6/E7 themselves or their interacting cellular proteins.

In 2014, a fatal infection with Legionella pneumophila serogroup 1 occurred in a neonate after a water birth. The death highlighted the need for infection control education, client awareness, and standardization of cleaning procedures in Texas midwife facilities.

During the 2012 outbreak of West Nile virus in the United States, approximately one third of the cases were in Texas. Of those, about half occurred in northern Texas. Models based on infected blood donors and persons with neuroinvasive disease showed, respectively, that ≈0.72% and 1.98% of persons in northern Texas became infected.

The RAS-RAF-MEK-ERK pathway is a major player in initiation and progression of multiple cancers where it can be hyperactivated by upstream regulatory proteins, such as tyrosine kinases, phosphatases, and GTPases. Several inhibitors targeting the RAS-MAPK pathway exhibit anticancer activity and are approved as single agent in specific cancers. One such is the MEK inhibitor trametinib, which is included as a rational polytherapy strategy for treating EML4-ALK, EGFR, and K-RAS mutant lung cancer and neuroblastoma containing hyperactivating RAS-MAPK pathway mutations. In neuroblastoma, a heterogeneous disease, relapse cases display an increased rate of mutations of ALK, NRAS, and NF1 genes, leading to hyperactivation of RAS-MAPK signaling. Targeting the RAS-MAPK pathways offers attractive options for combinatorial treatment together with ALK inhibitors, since monotreatment has not yet produced strong clinical results in ALK-positive neuroblastoma patients. Here we investigate the response of ALK-positive neuroblastoma cell lines to MEK inhibition, employing trametinib. We show that pharmacologic inhibition of the MEK-ERK pathway in ALK-positive neuroblastoma cells results in increased levels of activation/phosphorylation of AKT and ERK5. This feedback response is regulated by the mTORC2 complex protein SIN1. Taken together, our results indicate that blocking MEK-ERK leads to increased activation of AKT signaling core in ALK-positive neuroblastoma, intensifying survival signals in these cells. Our results contraindicate use of MEK inhibitors as an effective single- and polytherapeutic strategy in ALK-positive neuroblastoma. Citation Format: Bengt Hallberg, Ganseh Umapathy, Jikui Guan, Dan Gustafsson, Niloufar Javanmardi, Diana Cervantes-Madrid, Anna Djoos, Tommy Martinsson, Ruth Palmer. Anaplastic lymphoma kinase addicted neuroblastoma cell lines are associated with growth upon treatment with MEK inhibitor trametinib [abstract]. In: Proceedings of the AACR Special Conference: Pediatric Cancer Research: From Basic Science to the Clinic; 2017 Dec 3-6; Atlanta, Georgia. Philadelphia (PA): AACR; Cancer Res 2018;78(19 Suppl):Abstract nr B27.

Introduction Receptor tyrosine kinases (RTKs) play important roles in cellular proliferation and differentiation. In addition, RTKs reveal oncogenic potential when their kinase activities are constitutively enhanced by point mutation, amplification, or rearrangement of the corresponding genes. We are investigating ALK (Anaplastic Lymphoma Kinase) tyrosine kinase in neuroblastoma (NB) - the most frequently occurring extracranial tumour in children, where ALK activating mutations are found. In this study we performed phosphoproteomic and a gene signature profiling of ALK activity from several different neuroblastoma cells comparing treatment with the first and third generations ALK inhibitors. Material and methods Crizotinib was one of the first ALK TKIs to be described; however the response in patients with ALK positive NB was poor. In contrast, robust and sustained clinical responses we seen in ALK rearranged IMT. Lorlatinib is a third generation ALK TKI with very positive responses in NSCLC. We treated two ALK addicted (CLB-BAR, CLB-GE) and one ALK non-addicted (SK-N-AS) NB cell lines with either crizotinib or lorlatinib and performed phosphoproteomic and RNAseq profiling to identify common targets for future combinational treatment for patients. Results and discussions Profiling of ALK addicted CLB-BAR and CLB-GE cell lines led to the identification of 3345 and 2252 phosphoproteins, respectively. Phosphorylation of the ALK receptor itself was decreased in both lines upon treatment with crizotinib or lorlatinib. Parallel RNAseq profiling was performed at 24 hours. Comparison with non-treated controls revealed 19 232 differently expressed genes. In ALK addicted cells phosphoproteomic analysis revealed more than 50 proteins that were dephosphorylated upon treatment with ALK TKIs. More than 400 genes were downregulated, and more than 600 were upregulated, upon treatment with either crizotinib or lorlatinib. As an outcome we obtained a list of predicted targets for further analysis. We focused on validation of downstream signalling molecules such as: FOXO3a/4, DUSP4 and ETV3/4. Conclusion In this study we established and compared phosphoproteomic and gene signature profiles of ALK activity in NB cells employing first and third generation ALK TKIs. This analysis has unveiled a number of leads for novel combinatorial treatment strategies for NB patients as well as an increased understanding ALK dependant signalling processes.

Abnormal metabolism is another cancer hallmark. The two most characterized altered metabolic pathways are high rates of glycolysis and glutaminolysis, which are natural targets for cancer therapy. Currently, a number of newer compounds to block glycolysis and glutaminolysis are being developed; nevertheless, lonidamine and 6-diazo-5-oxo-L-norleucine (DON) are two old drugs well characterized as inhibitors of glycolysis and glutaminolysis, respectively, whose clinical development was abandoned years ago when the importance of cancer metabolism was not fully appreciated and clinical trial methodology was less developed. In this review, a PubMed search using the words lonidamine and 6-diazo-5-oxo-L-norleucine (DON) was undertaken to analyse existing information on the preclinical and clinical studies of these drugs for cancer treatment. Data show that they exhibit antitumor effects; besides there is also the suggestion that they are synergistic. We conclude that lonidamine and DON are safe and potentially effective drugs that need to be reevaluated in combination as metabolic therapy of cancer.