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Cancer is considered one of the leading causes of death in the United States. Although preventive strategies, early detection, and improved treatment options have been developed, novel targets and therapeutics are still needed. Since concluding that cancer is mediated by genetic and epigenetic alterations of the cell, many research groups are now focusing on other means of prevention and therapy via nutrition, epigenetic mechanisms, and non-coding RNAs which have been shown to control gene expression and have many different functions at the cellular level. With the advent of high-throughput sequencing in human cancer, the potential to identify novel biomarkers and therapeutic targets of disease has increased tremendously and led to the identification of many non-coding RNAs that are dysregulated in various cancers. Gene expression and regulation is important in maintaining the homeostasis of normal tissues and cells. Not uncommonly, up- or down-regulation of particular genes are associated with cancer as a result of increased or decreased expression of transcriptional targets. This review focuses on the role of nutrition in cancer and the dysregulation of non-coding RNAs with particular emphasis on long non-coding RNAs and microRNAs in different cancer types.

Alternative splicing (AS), a tightly regulated process, contributes to proteome diversification essential for tissue development and identity. During brain development, splicing targets include genes involved in several cellular processes like endocytosis, cytoskeleton dynamics and vesicle-mediated transport. Deregulation of AS in cancers enables tumor cells to alter the transcriptome in favor of isoforms that drive tumor progression. Annexin A7 (ANXA7) is a hydrophilic protein that binds membranes in a calcium-dependent manner; it is important for membrane scaffolding and vesicle trafficking. ANXA7 is alternatively spliced by PTBP1 and expressed as two isoforms - unspliced ANXA7 Isoform 1 (I1), containing cassette exon 6 and spliced Isoform 2 (I2). We determined that this splicing of ANXA7 is lineage-specific in the brain - I1 is highly expressed in post-mitotic mature neurons while I2 is expressed in neural and glial precursors. In order to understand the functional impact of these isoforms, we generated GBM cell lines that express endogenous I2 (I2 cells), or endogenous I2 + exogenous I1 (I1 cells). We evaluated the influence of I1 and/or I2 expression on RTK levels, activation and downstream events using western blot and co-immunoprecipitation assays. Receptor trafficking was observed using flow cytometry and immunofluorescence assays tagging different components of the endocytic pathway. In GBM, we observed high levels of PTBP1 with a subsequent increase in I2 levels indicative of dysregulated AS. Consequently, we observed upregulated activation of several receptor tyrosine kinases (RTKs) such as EGFR, MET, PDGFRα and EGFR vIII in the I2 cells. In I1 cells, however, we observed reduced levels and activation of all aforementioned RTKs along with the diminished activation of downstream pathways. Using EGFR signaling as a model, we found that in I1 cells EGFR co-localized with markers of lysosomal degradation indicating efficient trafficking of activated EGFR for endosomal degradation; co-localization was absent in I2 cells indicative of impaired trafficking. We propose that I1, by virtue of the amino acids encoded by cassette exon 6, binds to conserved motifs within multiple RTKs and targets them for endosomal degradation thereby attenuating tumorigenic signaling in GBM. Our results show that ANXA7 I1 is tumor suppressive in GBM and that loss of I1 promotes tumorigenicity by perpetuating RTK signaling. We present a mechanism that explains, in part, how I1 mediates the degradation of multiple tumorigenic RTKs by spatio-temporally regulating their endosomal trafficking. A refined understanding of how I1 functions will provide the foundation for future, selective therapies that target dysregulated AS in GBM. Citation Format: Sindhu Nair, Rajani Rajbhandari, Centdrika Dates-Hurt, Markus Bredel. Dysregulated splicing of Annexin A7 impairs receptor tyrosine kinase trafficking in glioblastoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3747.

Purpose/Objective(s): Glioblastomas (GBM) are highly malignant tumors that arise from astrocytes. GBMs rely on signaling through receptor tyrosine kinases (RTK) to ensure tumorigenicity. EGFR, PDGFRA and MET are distinct RTKs; each is capable of binding a ligand, which promotes receptor dimerization and kinase activation. Annexin A7 (ANXA7), a protein involved in membrane binding and vesicle trafficking, is spliced into 2 isoforms - isoforms 1 (I1) and isoform 2 (I2). A normal adult brain will express both isoforms, however, in GBM, I1 expression is prevented. We determined that I1, but not I2, is tumor suppressive, in part, because it promotes the endocytic degradation of EGFR. Because I1 is typically not expressed in GBM, EGFR signaling persists unabated and tumors thrive. Recently, we extended these observations and determined that I1 also impacts additional RTKs, including PDGFRA and MET. By understanding how ANXA7 I1 impacts PDGFRA and MET signaling, we hope to target multiple tumorigenic RTKs and improve therapy for patients with GBM. Materials/Methods: To understand how and when I1 downregulates PDGFRα and MET, U251-malignant glioma (MG) cells, which only express I2 (P), were modified to express I1. Cells were serum starved overnight, and then stimulated with PDGFAA and HGF(20 ng/ml), which activate the aforementioned receptors respectively. As a positive control, cells were stimulated with EGF to activate EGFR. Total protein was then collected and analyzed by western blot analyses. Next, we sought to determine whether I1 was competent to down-regulate several RTKs coincidentally. To do this, P and I1 cells were serum starved, stimulated with EGF, PDGFAA or both, and total protein levels were evaluated. Results: The levels of activated EGFR, PDGFRα and MET are dramatically reduced in those cells that express I1 as compared to cells expressing I2 (P). Upon activation with both PDGFAA and EGF, in cells expressing I1 (I1), levels of PDGFRα and EGFR were markedly reduced 30 minutes post-stimulation as compared to cells expressing I2. Conclusion: Our results suggest I1 may be a master regulator of RTK endocytosis and degradation. Further investigations of the impact of I1 on endocytic trafficking patterns using immunofluorescence and confocal microscopy are underway. Collectively, understanding how I1 functions and targets multiple tumorigenic RTKs will provide the foundation for future selective therapies that restore I1 expression in GBM and reduce tumorigenicity. Citation Format: Zachary B. White, Centdrika Dates Hurt, Rajani Rajbhandari, Sindhu Nair, Susan Nozell, Markus Bredel. Impact of ANXA7 I1 expression on PDGFRA and MET endosomal trafficking in glioblastoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3758.

Dysregulation of receptor tyrosine kinase genes is a frequent genetic abnormality in GBM. A common genetic alteration is amplification and/or overexpression of the epidermal growth factor receptor (EGFR) gene. Annexin A7 (ANXA7) is a hydrophilic protein that binds membranes in a calcium dependent manner; it is important for membrane scaffolding and vesicle trafficking. ANXA7 is alternatively spliced and expressed as two isoforms – unspliced ANXA7 Isoform 1 (I1), containing cassette exon 6 and spliced Isoform 2 (I2). Our lab has determined that I1 is tumor suppressive in GBM and that loss of I1 promotes tumorigenicity by perpetuating EGFR signaling. However, if restored, I1 but not I2, reduces EGFR levels, activation and downstream pathways and diminishes tumorigenicity. In patients, I1 levels directly correlate with survival and prognosis. We propose that I1 inhibits EGFR signalling by binding to and promoting degradation of EGFR in a calcium dependent manner. In GBM, I1 is absent, preventing endosomal degradation of EGFR and leading to perpetual signalling. Our approach is novel; rather than address how/whether EGFR is activated, we focus on why EGFR fails to be downregulated afterwards. Herein, we present a mechanism that explains, in part, how I1 mediates endosomal degradation of EGFR. Further studies are underway to identify exactly where and how I1 modifies endosomal trafficking and degradation of EGFR. We are also exploring the role of I1 in regulating the degradation of other receptor tyrosine kinases (RTKs), such as MET and PDGFRA. Collectively, understanding how I1 functions will provide the foundation for future, selective therapies that restore I1 expression and/or function to reduce GBM tumorigenicity.

Background: Funded by the Third EU Public Health Programme, Optimity Advisors has followed up on the work carried out by the Health System Performance Assessment HSPA Expert Group in the development of an assessment framework for integrated care. The study conducted has assessed the level penetration and adoption of integrated care in the EU28, Norway and Iceland, and the level of maturity of several health systems at national, regional and local level. Through engagement with stakeholders in the integrated care field, a new, user-friendly framework for the performance assessment of integrated care has been co-developed and further validated by Optimity Advisors, DG SANTE i.e. Directorate-General for Health and Consumers of the European Union, CHAFEA Consumers, Health, Agriculture and Food Executive Agency and stakeholders across the integrated care community in the 28 Member States, Norway and Iceland. The Integrated Care Performance Assessment Framework was developed based on previously validated frameworks, such as the WHO 2015 framework People-centred and integrated health services: an overview of the evidence. Aims and Objectives: The main objective of this workshop will be to communicate the preliminary results and the experience of using the framework to assess the performance of a number of integrated care systems at local, regional and national level. Format: The agenda for the workshop will be as follows: Introduction to the methodological approach and preliminary outputs of the framework, by Optimity Advisors and DG SANTE; Presentation by one or two representatitves of integrated care projects who have contributed in the co-design of the framework explaining the design process; Demonstration of the use of the framework by representatitves of integrated care projects, who will show how to obtain a performance profile for integrated care systems; Group discussion on the different conditions in different health systems that can enhance, or limit the adoption of the framework, and on what could be done to minimise the barriers and maximise facilitating conditions; Conclusion by the facilitators on future steps to address the issues raised by the attendees, and final remarks. Target audience: We envision the target audience of the workshop to be comprised of individuals who have an integrated care performance assessment need, including health managers, policy makers, and other stakeholders that are directly involved in decision-making actions within integrated health systems. Learnings/Take away: By the end of the session, we expect attendees to i have an understanding of a new way to assess the performance of integrated care systems, ii develop a performance profile and actionable roadmap for performance improvement of an integrated care system of their choosing.

Increased aerobic glycolysis and de novo lipid biosynthesis are common characteristics of invasive cancers. UDP-glucuronosyltransferases (UGTs) are phase II drug metabolizing enzymes that in normal cells possess the ability to glucuronidate these lipids and speed their excretion; however, de-regulation of these enzymes in cancer cells can lead to an accumulation of bioactive lipids, which further fuels cancer progression. We hypothesize that UGT2B isoform expression is down-regulated in cancer cells and that exogenous re-introduction of these enzymes will reduce lipid content, change the cellular phenotype, and inhibit cancer cell proliferation. In this study, steady-state mRNA levels of UGT isoforms from the 2B family were measured using qPCR in 4 breast cancer and 5 pancreatic cancer cell lines. Expression plasmids for UGT2B isoforms known to glucuronidate cellular lipids, UGT2B4, 2B7, and 2B15 were transfected into MCF-7 and Panc-1 cells, and the cytotoxic effects of these enzymes were analyzed using trypan blue exclusion, annexin V/PI staining, TUNEL assays, and caspase-3 immunohistochemistry. There was a significant decrease in cell proliferation and a significant increase in the number of dead cells after transfection with each of the 3 UGT isoforms in both cell lines. Cellular lipids were also found to be significantly decreased after transfection. The results presented here support our hypothesis and emphasize the important role UGTs can play in cellular proliferation and lipid homeostasis. Evaluating the effect of UGT expression on the lipid levels in cancer cell lines can be relevant to understanding the complex regulation of cancer cells, identifying the roles of UGTs as “lipid-controllers” in cellular homeostasis, and illustrating their suitability as targets for future clinical therapy development.

Tamoxifen (Tam) is a selective estrogen receptor modulator used to inhibit breast tumor growth. Tam can be directly N-glucuronidated via the tertiary amine group or O-glucuronidated after cytochrome P450–mediated hydroxylation. In this study, the glucuronidation of Tam and its hydroxylated and/or chlorinated derivatives [4-hydroxytamoxifen (4OHTam), toremifene (Tor), and 4-hydroxytoremifene (4OHTor)] was examined using recombinant human UDP-glucuronosyltransferases (UGTs) from the 1A subfamily and human hepatic microsomes. Recombinant UGT1A4 catalyzed the formation of N-glucuronides of Tam and its derivatives and was the most active UGT enzyme toward these compounds. Therefore, it was hypothesized that single nucleotide polymorphisms (SNPs) in the promoter region of UGT1A4 have the ability to significantly decrease the glucuronidation rates of Tam metabolites in the human liver. In vitro activity of 64 genotyped human liver microsomes was used to determine the association between the UGT1A4 promoter and coding region SNPs and the glucuronidation rates of Tam, 4OHTam, Tor, and 4OHTor. Significant decreases in enzymatic activity were observed in microsomes for individuals heterozygous for −163G/A and −217T/G. These alterations in glucuronidation may lead to prolonged circulating half-lives and may potentially modify the effectiveness of these drugs in the treatment of breast cancer.

Latinos constitute the largest ethnic minority group in the United States. However, the cultural adaptation and dissemination of evidence-based parenting interventions among Latino populations continues to be scarce despite extensive research that demonstrates the long-term positive effects of these interventions. The purpose of this article is threefold: (1) justify the importance of cultural adaptation research as a key strategy to disseminate efficacious interventions among Latinos, (2) describe the initial steps of a program of prevention research with Latino immigrants aimed at culturally adapting an evidence-based intervention informed by parent management training principles, and (3) discuss implications for advancing cultural adaptation prevention practice and research, based on the initial feasibility and cultural acceptability findings of the current investigation.

Tuberculosis (TB) and Human Immunodeficiency Virus (HIV) are two catastrophic diseases affecting millions of people worldwide every year, and are considered to be pandemic by the World Health Organization. This study aims to compare the recent trends in TB and HIV in the United States and Sub-Saharan African Countries. Data (incidence, prevalence and death rates of HIV and TB) for the United States, Cameroon, Nigeria, and South Africa were collected from The Joint United Nations Programme for HIV/AIDS (UNAIDS), US Census Bureau and World Health Organization (WHO) databases and analyzed using Statistical Analysis Software (SAS v 9.1). Analysis of Variance (ANOVA) was performed to compare the variables of interest between the countries and across time. Results showed that percent rates of TB cases, TB deaths, HIV cases and HIV deaths were significantly different (P P P < 0.001) in the prevalence of TB and HIV between the United States and the Sub-Saharan African countries, as well as differences within the Sub-Saharan African countries from 1993 to 2006. More analysis needs to be carried out in order to determine the prevalence and incidence of HIV and TB among multiple variables like gender, race, sexual orientation and age to get a comprehensive picture of the trends of HIV and TB.

The Assertive Community Treatment model (ACT) was developed more than 30 years ago to treat individuals with serious and persistent mental illness. This qualitative study highlights practical challenges encountered when establishing ACT teams in diverse settings serving people who are homeless with co-occurring mental and addictive disorders (COD). Program administrators and evaluators from nine programs located in seven states completed a survey on implementation challenges, fidelity, modifications to the ACT model, and program successes. Challenges encountered related largely to staffing and funding limitations as well as to difficulties with implementing the ACT model without modifications. Several modifications to the model were believed beneficial to recruiting and retaining consumers. These included emphasizing housing, adding staff positions not prescribed by ACT, implementing mini-teams within the program, delivering in-office services in a group format, and placing time-limited services by transi...

An estimated 3.5 million people in the United States meet criteria for being homeless (National Law Center on Homelessness & Poverty, 2007), about 1.4% to 2% of the population (Greenberg & Rosenhec...

Tamoxifen (Tam) is a selective estrogen receptor (ER) modulator used to inhibit breast tumor growth. Tam can be directly N-glucuronidated via the tertiary amine group or O-glucuronidated after cytochrome P450 mediated hydroxylation. In this study, the glucuronidation of Tam and its hydroxylated and/or chlorinated derivatives [4OHTamoxifen (4OHTam), Toremifene (Tor), and 4OHTor] was examined using recombinant human UGTs from the 1A subfamily and human hepatic microsomes. Recombinant UGT1A4 catalyzed the formation of N-glucuronides of Tam and its derivatives and was the most active UGT enzyme toward these compounds. Therefore, it was hypothesized that single nucleotide polymorphisms (SNPs) in the promoter region of UGT1A4 have the ability to significantly decrease the glucuronidation rates of Tam metabolites in the human liver. In vitro activity of 64 genotyped human liver microsomes were used to determine the association between the UGT1A4 promoter and coding region SNPs and the glucuronidation rates of Tam, 4OHTam, Tor, and 4OHTor. Significant decreases in enzymatic activity were observed in microsomes for individuals heterozygous for -163G/A and -217T/G. These alterations in glucuronidation may lead to prolonged circulating half-lives and potentially modify the effectiveness of these drugs in the treatment of breast cancer.

Tamoxifen (Tam) is classified as a selective estrogen receptor modulator (SERM) and is used for treatment of patients with ER-positive breast cancer. However, it has been shown that Tam and its cytochrome P450-generated metabolite 4-hydroxy-Tam (4OH-Tam) also exhibit cytotoxic effects in ER-negative breast cancer cells. These observations suggest that Tam and 4OH-Tam can produce cytotoxicity via estrogen receptor (ER)-independent mechanism(s) of action. The molecular targets responsible for the ER-independent effects of Tam and its derivatives are poorly understood. Interestingly, similar to Tam and 4OH-Tam, cannabinoids have also been shown to exhibit anti-proliferative and apoptotic effects in ER-negative breast cancer cells, and estrogen can regulate expression levels of cannabinoid receptors (CBRs). Therefore, this study investigated whether CBRs might serve as novel molecular targets for Tam and 4OH-Tam. We report that both compounds bind to CB1 and CB2Rs with moderate affinity (0.9-3 μM). Furthermore, Tam and 4OH-Tam exhibit inverse activity at CB1 and CB2Rs in membrane preparations, reducing basal G-protein activity. Tam and 4OH-Tam also act as CB1/CB2R-inverse agonists to regulate the downstream intracellular effector adenylyl cyclase in intact cells, producing concentration-dependent increases in intracellular cAMP. These results suggest that CBRs are molecular targets for Tam and 4OH-Tam and may contribute to the ER-independent cytotoxic effects reported for these drugs. Importantly, these findings also indicate that Tam and 4OH-Tam might be used as structural scaffolds for development of novel, efficacious, non-toxic cancer drugs acting via CB1 and/or CB2Rs.

Objetivos. Determinar la seroprevalencia de la infeccion por Trypanosoma cruzi en Ushuaia, la ciudad mas austral del mundo. Metodos. Se analizaron muestras de suero de 2 991 personas, obtenidas entre enero de 1995 y diciembre de 1996. Las muestras fueron procesadas por hemaglutinacion indirecta (HAI) e inmunoensayo enzimatico (ELISA) o inmunofluorescencia indirecta (IFI). Resultados. La seroprevalencia general de la infeccion por T. cruzi fue de 6,8%. La prevalencia segun el pais de origen fue de 41,1% en los bolivianos, 5,0% en los argentinos y 0,9% en los chilenos; en embarazadas fue de 5,9%, en examenes obligatorios de 6,3% y en consultas dirigidas de 30,8%. Conclusiones. Se destaca la magnitud de la infeccion por T. cruzi en una zona donde no existe el insecto vector. Debido al riesgo de la transmision congenita y transfusional, es necesario mantener el control de la sangre a transfundir y reforzar el seguimiento de los hijos de mujeres infectadas para un diagnostico precoz y tratamiento oportuno de la infeccion.

This study examined the association between ownership of high-risk (“vicious”) dogs and the presence of deviant behaviors in the owners as indicated by court convictions. We also explored whether two characteristics of dog ownership (abiding licensing laws and choice of breed) could be useful areas of inquiry when assessing risk status in settings where children are present. Our matched sample consisted of 355 owners of either licensed or cited dogs that represented high or low-risk breeds. Categories of criminal convictions examined were aggressive crimes, drugs, alcohol, domestic violence, crimes involving children, firearm convictions, and major and minor traffic citations. Owners of cited high-risk (“vicious”) dogs had significantly more criminal convictions than owners of licensed low-risk dogs. Findings suggest that the ownership of a high-risk (“vicious”) dog can be a significant marker for general deviance and should be an element considered when assessing risk for child endangerment.

​FormerFormer consultant ophthalmic surgeon Salisbury Infirmary and Odstock Hospital, Wiltshire (b 1921; q Oxford/The London Hospital 1946; DO, FRCS), d 19 February 2005. Figure 1 After house jobs and service in the Royal Navy John held ophthalmic posts in Oxford and at Moorfields Hospital, where he became senior registrar. He was appointed to Salisbury in 1955, where he remained for the rest of his career. John was a member of the Magic Circle, and a skilled carpenter and joiner, who designed and made miniature dolls houses and automata (the picture shows him with a medieval cathedral builder, part of an enormous model that he made of Salisbury Cathedral). His great love was the restoration of a near derelict croft on the Hebridean island of Coll. John published House in the Hebrides in 2004 about the croft's restoration and associated adventures. He leaves Doreen, four children, and six grandchildren.

The consequences of aging of the immune system include impaired T-lymphocyte responsiveness and aberrant immunoglobulin production. Although T cells from elderly individuals have a well-described defect in lymphoblastic transformation in response to some polyclonal mitogens, immunoglobulin abnormalities have lacked a clear in vitro model. Peripheral blood mononuclear cells from 13 young and 13 old healthy donors were cultured with phytohemagglutinin (PHA) or pokeweed mitogen (PWM). Old-donor-cell phytohemagglutinin (PHA), but not PWM, cultures had significantly lower lymphoblastic transformation compared with young donor cultures. IgG, IgA, and IgM production tended to be lower in old- versus young-donor PWM cell cultures. By contrast, despite lower lymphoblastic transformation in old-donor PHA cell cultures, immunoglobulin production was higher for old- versus young-donor cell cultures. No significant age differences were present in initial lymphocyte counts, percent B cells, T cells or monocytes, or helper/suppressor ratios to explain this enhancement in immunoglobulin production. PHA-stimulated mononuclear cell cultures in the aged demonstrate not only a defect in proliferation but also increased immunoglobulin production. This in vitro system may be useful to characterize further the pathogenesis of altered immunoglobulin production in the elderly.

Drosophila melanogaster larvae require choline for growth but when fed dl-carnitine in place of choline, carnitine is decarboxylated to β-methylcholine and phosphatidyl-β-methylcholine is synthesized to replace phosphatidylcholine. Feeding (−)-carnitine in place of choline results in a moderate deficiency of lecithin-type phospholipid and a corresponding reduction in growth. The capacity of larvae to employ (+)-carnitine for phosphatidyl-β-methylcholine synthesis and subsequently for growth is very limited. Only 41% as much phosphatidyl-β-methylcholine is synthesized by larvae fed 5.7 X 10−4 M (+)-carnitine as is formed when the same concentration of (−)-carnitine is fed. This suggests that (−)-carnitine is the preferred substrate for carnitine decarboxylase, the enzyme that degrades carnitine to form β-methylcholine. Utilization of carnitine for phospholipid synthesis is clearly dependent upon a choline-deficient state. The presence of small quantities of choline, either in body tissues or in the diet, inhibits the incorporation of carnitine derivatives into phospholipid. Intact carnitine is needed for maximum larval growth and adult activity, indicating that D. melanogaster larvae synthesize a suboptimal quantity of carnitine. Thus, dietary carnitine supplements in vivo carnitine synthesis for physiological purposes.