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Supplementary Figure S1. Dose-response curves of combinations of orlistat or NanoOrl with taxanes; Supplementary Figure S2. Synergy analysis with Combenefit (Loewe model);Supplementary Figure S3. Synergy analysis with Combenefit [Highest Single Agent (HSA) model];Supplementary Figure. S4. Morphological changes associated with combination treatment;Supplementary Figure S5. Combination of NanoOrl plus docetaxel stabilizes microtubules in taxane-resistant cells;Supplementary Fig. S6. Example of overestimation of synergy; Supplementary Fig. S7. Example of underestimation of synergy.

This study examined the role of demographics, civic beliefs, and the impact of the COVID-19 pandemic in association with distinct forms of civic participation. College students were recruited across 10 institutions of higher education to complete an online survey. Bivariate, multivariable linear, and logistic regressions were performed. Findings indicated that participants from traditionally marginalized backgrounds were more likely to engage in systemchallenging forms of civic participation and community engagement than those from more privileged backgrounds. Participants who rated high in critical reflection, viewed racism as a key issue, and were heavily impacted by the COVID-19 pandemic were also more likely to engage in system-challenging forms of civic participation. Participants who endorsed beliefs supporting current systems of power were more likely to report they intended to vote. Results highlight implications for antiracist activism, community engagement, and traditional political civic behaviors.

Youth who report prosocial relationships with natural mentors show increased social and academic success and life satisfaction. However, little research investigates how mentors impact aspects of youth identity such as athleticism or religiosity. The present study applies a Bayesian Additive Regression Trees (BART) analysis model to data from a national longitudinal study ( N = 15,701) to predict specific outcomes in educational, athletic, and religious domains. This analytic approach is uniquely well-suited for accurately drawing conclusions within highly collinear, high dimensional datasets. Analyses included demographic variables and childhood base rates of academic success, fitness, and religious beliefs as covariates. Findings indicate that the presence of an academic mentor during adolescence longitudinally predicts educational attainment, while athletic or religious mentors did not have significant impacts in terms of increasing athleticism or religiosity. These results suggest that academic mentors may have more longitudinal impacts on student success than other types of mentors.

In the United States, schools provide a large portion of child and adolescent mental health services; however, systems are restricted by resource and workforce shortages while the need for services...

The preference for and exercise of autonomous decision-making in adolescence is a normative developmental process. Yet, increased autonomy is associated with both risks and benefits. Connection to others through positive relationships, including mentoring relationships, is one context that predicts healthy autonomous decision-making. In other ways, such relationships can interfere or stifle the development of autonomy. In synthesizing the existing scientific literature on autonomy development and autonomy-supportive practices, we propose a framework for considering the role of mentors in supporting autonomy through five domains of influence: role modeling, encouraging, providing access to resources, relationships, and experiences, advocacy, and conversations about behavior change. We provide suggestions for research and practice.

Tumor cells require increased rates of cell metabolism to generate the macromolecules necessary to sustain proliferation. They rely heavily on NAD+ as a cofactor for multiple metabolic enzymes in anabolic and catabolic reactions. NAD+ also serves as a substrate for PARPs, sirtuins, and cyclic ADP-ribose synthases. Dysregulation of the cyclic ADP-ribose synthase CD38, the main NAD'ase in cells, is reported in multiple cancer types. This study demonstrates a novel connection between CD38, modulation of NAD+, and tumor cell metabolism in prostate cancer. CD38 expression inversely correlates with prostate cancer progression. Expressing CD38 in prostate cancer cells lowered intracellular NAD+, resulting in cell-cycle arrest and expression of p21Cip1 (CDKNA1). In parallel, CD38 diminishes glycolytic and mitochondrial metabolism, activates AMP-activated protein kinase (AMPK), and inhibits fatty acid and lipid synthesis. Pharmacologic inhibition of nicotinamide phosphoribosyltransferase (NAMPT) mimicked the metabolic consequences of CD38 expression, demonstrating similarity between CD38 expression and NAMPT inhibition. Modulation of NAD+ by CD38 also induces significant differential expression of the transcriptome, producing a gene expression signature indicative of a nonproliferative phenotype. Altogether, in the context of prostate cancer, the data establish a novel role for the CD38–NAD+ axis in the regulation of cell metabolism and development. Implications: This research establishes a mechanistic connection between CD38 and metabolic control. It also provides the foundation for the translation of agents that modulate NAD+ levels in cancer cells as therapeutics. Mol Cancer Res; 16(11); 1687–700. ©2018 AACR.

Taxane-based therapy provides a survival benefit in patients with metastatic prostate cancer, yet the median survival is less than 20 months in this setting due in part to taxane-associated resistance. Innovative strategies are required to overcome chemoresistance for improved patient survival. Here, NanoOrl, a new experimental nanoparticle formulation of the FDA-approved drug, orlistat, was investigated for its cytotoxicity in taxane-resistant prostate cancer utilizing two established taxane-resistant (TxR) cell lines. Orlistat is a weight loss drug that inhibits gastric lipases, but is also a potent inhibitor of fatty acid synthase (FASN), which is overexpressed in many types of cancer. NanoOrl was also investigated for its potential to synergize with taxanes in TxR cell lines. Both orlistat and NanoOrl synergistically inhibited cell viability when combined with paclitaxel, docetaxel, and cabazitaxel in PC3-TxR and DU145-TxR cells, yet these combinations were also additive in parental lines. We observed synergistic levels of apoptosis in TxR cells treated with NanoOrl and docetaxel in combination. Mechanistically, the synergy between orlistat and taxanes was independent of effects on the P-glycoprotein multidrug resistance protein, as determined by an efflux activity assay. On the other hand, immunoblot and immunofluorescence staining with an anti-detyrosinated tubulin antibody demonstrated that enhanced microtubule stability was induced by combined NanoOrl and docetaxel treatment in TxR cells. Furthermore, TxR cells exhibited higher lipid synthesis, as demonstrated by 14C-choline incorporation that was abrogated by NanoOrl. These results provide a strong rationale to assess the translational potential of NanoOrl to overcome taxane resistance. Mol Cancer Ther; 16(9); 1819–30. ©2017 AACR.

Fatty acid synthase (FASN), the enzyme that catalyzes de novo synthesis of fatty acids, is expressed in many cancer types. Its potential as a therapeutic target is well recognized, but inhibitors of FASN have not yet been approved for cancer therapy. Orlistat (ORL), an FDA-approved lipase inhibitor, is also an effective inhibitor of FASN. However, ORL is extremely hydrophobic and has low systemic uptake after oral administration. Thus, new strategies are required to formulate ORL for cancer treatment as a FASN inhibitor. Here, we report the development of a nanoparticle (NP) formulation of ORL using amphiphilic bioconjugates that are derived from hyaluronic acid (HA), termed Nano-ORL. The NPs were loaded with up to 20 wt % weight of ORL at greater than 95% efficiency. The direct inhibition of the human recombinant thioesterase domain of FASN by ORL extracted from Nano-ORL was similar to that of stock ORL. Nano-ORL demonstrated a similar ability to inhibit cellular FASN activity when compared to free ORL, as demonstrated by analysis of (14)C-acetate incorporation into lipids. Nano-ORL treatment also disrupted mitochondrial function similarly to ORL by reducing adenosine triphosphate turnover in MDA-MB-231 and LNCaP cells. Nano-ORL demonstrated increased potency compared to ORL toward prostate and breast cancer cells. Nano-ORL decreased viability of human prostate and breast cancer cell lines to 55 and 57%, respectively, while free ORL decreased viability to 71 and 79% in the same cell lines. Moreover, Nano-ORL retained cytotoxic activity after a 24 h preincubation in aqueous conditions. Preincubation of ORL dramatically reduced the efficacy of ORL as indicated by high cell viability (85%) in both breast and prostate cell lines. These data demonstrate that NP formulation of ORL using HA-derived polymers retains similar levels of FASN, lipid synthesis, and ATP turnover inhibition while significantly improving the cytotoxic activity against cancer cell lines.

Living shorelines are a type of estuarine shoreline erosion control that incorporates native vegetation and preserves native habitats. Because they provide the ecosystem services associated with natural coastal wetlands while also increasing shoreline resilience, living shorelines are part of the natural and hybrid infrastructure approach to coastal resiliency. Marshes created as living shorelines are typically narrow (< 30 m) fringing marshes with sandy substrates that are well flushed by tides. These characteristics distinguish living shorelines from the larger meadow marshes in which most of the current knowledge about created marshes was developed. The value of living shorelines for providing both erosion control and habitat for estuarine organisms has been documented but their capacity for carbon sequestration has not. We measured carbon sequestration rates in living shorelines and sandy transplanted Spartina alterniflora marshes in the Newport River Estuary, North Carolina. The marshes sampled here range in age from 12 to 38 years and represent a continuum of soil development. Carbon sequestration rates ranged from 58 to 283 g C m-2 yr-1 and decreased with marsh age. The pattern of lower sequestration rates in older marshes is hypothesized to be the result of a relative enrichment of labile organic matter in younger sites and illustrates the importance of choosing mature marshes for determination of long-term carbon sequestration potential. The data presented here are within the range of published carbon sequestration rates for S. alterniflora marshes and suggest that wide-scale use of the living shoreline approach to shoreline management may come with a substantial carbon benefit.

The selective α-reduction of N-protected acyl aziridines is achieved with methyldiphenylsilyllithium in THF to produce Mannich bases.

The majority of cancers undergo metabolic alterations during initiation and progression of disease. Increased de novo lipogenesis is recognized as metabolic mark of cancer cells. The enzymes responsible for de novo fatty acid synthesis are often overexpressed in cancer, indicating that these enzymes are potential and promising targets for novel therapies. Acetyl CoA Carboxylase 1 (ACC1) is a cytosolic enzyme which catalyzes the rate limiting step of de novo fatty acid synthesis through the ATP and biotin-dependent carboxylation of acetyl-CoA to malonyl-CoA. In doing so, it regulates both fatty acid and acetate metabolism. Both pharmacological inhibition and siRNA knockdown of ACC1 have been demonstrated to deny prostate cancer cells of necessary amounts of fatty acids needs to proliferate, resulting in selective toxicity. Although previous research indicates ACC1 and the entire process of fatty acid metabolism to be promising avenues for novel therapy development, much remains to be understood. To address this, we have generated mice with prostate-specific deletion of exon 22 of the Acetyl CoA Carboxylase 1 (ACCL/L) gene by breeding with Probasin-Cre mice. Loss of ACC1 activity does not affect prostate development as survival, prostate weight, histology, and breeding were similar between ACCL/L; Cre+ and ACCL/L Cre- mice. By breeding ACC+/L mice with the established Pten knockout model of prostate cancer (PtenL/L; Cre+), we have found that homozygous inactivation of ACC1 (ACCL/L; PtenL/L; Cre+) is able to reduce tumor burden in mice at 12 weeks of age when compared to ACC+/+; PtenL/L; Cre+ littermates (average total prostate weight of 90mg versus 120mg respectively, normalized to body weight, p0.05). In our efforts to detail why ACC1 inactivation dampens early tumor burden but not later cancer progression, we have found that tumors from both 12 and 24 week old ACCL/L; PtenL/L; Cre+ mice show no changes in lipid profile when compared to tumors arising from age-matched ACC+/+; PtenL/L; Cre+ siblings. Interestingly, through metabolic analysis, we have found that different prostate cancer cell lines utilize fatty acids in unique ways. Under conditions that mimic bioenergetic stress, only androgen insensitive DU145 cells, and not LNCaPs, were able to oxidize exogenous fatty acids. Moreover, treatment with the non-isoform specific ACC inhibitor TOFA (5-Tetradecyloxy-2-furoic acid) abolished fatty acid oxidation under such conditions. These data indicate a potential for metabolic rewiring associated with disease progression and/or metabolic stress, and that blockade of ACC1 may not be sufficient to prevent disease progression. The role of ACC1 in acetate metabolism, redox modulation, fatty acid oxidation, and compensation for ACC1 activity by ACC2 are all potential mechanisms by which ACC1-deficient prostate cancer cells are able to survive and proliferate. Ultimately, these data further the notion that tumors rewire their metabolic circuits to meet the demands associated with cancer biology. Citation Format: Amanda L. Davis, Kristen Scott, Wu Jiansheng, Yong Q. Chen, Steven J. Kridel. ACC1 is required for prostate cancer initiation, but not progression? [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A21.

Tks5 is a Src substrate and adaptor protein previously recognized for its regulation of cancer cell invasion through modulation of specialized adhesion structures called podosomes/invadopodia. Here we show for the first time that Tks5 localizes to the podosomes of primary macrophages, and that Tks5 protein levels increase concurrently with podosome deposition during the differentiation of monocytes into macrophages. Similar results are reported for model THP-1 cells, which differentiate into macrophages and form proteolytically active podosomes in response to a PKC signaling agonist (PMA) and with sensitivity to a PKC inhibitor (bisindolylmaleimide). Genetic manipulation of Tks5 expression (silencing and overexpression) in stable THP-1 cell lines does not independently alter this macrophage differentiation process. Nor do these cells lose the ability to focalize F-actin and its accessory proteins into podosome-like structures following PMA treatment. However, Tks5 directly controls podosome-associated gelatin degradation and invasion through collective changes in adhesion, chemotaxis, and the expression/proteolytic activity of MMP9. The Src family kinase-dependent phosphorylation of Tks5 is also implicated in the regulation of THP-1 macrophage invasive behavior. These results therefore define a previously unappreciated function of Tks5 signaling specific to the functional attributes of the macrophage podosome in adhesion, motility, and extracellular matrix-remodeling.

Mannich bases are prepared from the selective α-reduction of acyl aziridines using silyllithium reagents. The reaction proceeds via an aziridine ring-opening assisted Brook rearrangement.

The alteration of cellular metabolic processes is a common event in the initiation and progression of several cancer types. One metabolic hallmark of cancer cells is an increase in de novo lipogenesis. The enzymes responsible for de novo fatty acid synthesis are overexpressed in cancer, suggesting that these enzymes may be appropriate targets in efforts to inhibit or treat disease. Specifically, Acetyl CoA Carboxylase 1 (ACC1), the cytosolic enzyme that catalyzes the rate limiting step of de novo fatty acid synthesis through the ATP and biotin-dependent carboxylation of acetyl-CoA to malonyl-CoA, has received attention as a potential target for cancer treatment. Pharmacological inhibition and siRNA mediated knockdown of ACC1 has been shown to selectively kill prostate cancer cells in vitro by denying cells of adequate amounts of fatty acids needed to proliferate and survive. Although ACC1 has been implicated in cancer, its role has not been examined in a spontaneous model of cancer. To address this issue, we first generated mice with prostate-specific deletion of exon 22 of the Acetyl CoA Carboxylase 1 (ACCL/L) gene by breeding with Probasin-Cre mice. Loss of ACC1 activity did not appear to affect prostate function as survival, prostate weight, histology, and breeding was similar between ACCL/L Cre+ and ACCL/L Cre− mice. To determine the role of ACC1 in prostate cancer, the ACC+/L mice were crossed with PtenL/L; Cre+ mice. To our surprise, ACC1 activity was not required for prostate cancer development or progression in vivo, despite its apparent requirement in vitro. Total prostate weight and individual lobe weight did not differ among ACC+/L; PtenL/L; Cre+ mice, ACCL/L; PtenL/L; Cre+ mice and ACC1+/+; PtenL/L; Cre+ mice. These early studies suggest that ACC1 activity, and perhaps the fatty acid synthesis pathway, may not be required for prostate cancer development. The role of ACC1 in redox modulation, rescue of impaired de novo fatty acid synthesis by circulating and dietary fatty acid, and the potential compensation for ACC1 activity by ACC2 in prostate cancer are possible explanations for this unexpected phenotype. These results could have significant impact on the development and use of anti-cancer therapeutics that target the fatty acid synthesis pathway. Citation Format: Amanda L. Davis, Kristen Scott, Yong Chen, Steven Kridel. Acetyl coA carboxylase1 and its role in prostate cancer initiation and progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1878. doi:10.1158/1538-7445.AM2013-1878

Invadopodia are membrane protrusions that facilitate matrix degradation and cellular invasion. Although lipids have been implicated in several aspects of invadopodia formation, the contributions of de novo fatty acid synthesis and lipogenesis have not been defined. Inhibition of acetyl-CoA carboxylase 1 (ACC1), the committed step of fatty acid synthesis, reduced invadopodia formation in Src-transformed 3T3 (3T3-Src) cells, and also decreased the ability to degrade gelatin. Inhibition of fatty acid synthesis through AMP-activated kinase (AMPK) activation and ACC phosphorylation also decreased invadopodia incidence. The addition of exogenous 16∶0 and 18∶1 fatty acid, products of de novo fatty acid synthesis, restored invadopodia and gelatin degradation to cells with decreased ACC1 activity. Pharmacological inhibition of ACC also altered the phospholipid profile of 3T3-Src cells, with the majority of changes occurring in the phosphatidylcholine (PC) species. Exogenous supplementation with the most abundant PC species, 34∶1 PC, restored invadopodia incidence, the ability to degrade gelatin and the ability to invade through matrigel to cells deficient in ACC1 activity. On the other hand, 30∶0 PC did not restore invadopodia and 36∶2 PC only restored invadopodia incidence and gelatin degradation, but not cellular invasion through matrigel. Pharmacological inhibition of ACC also reduced the ability of MDA-MB-231 breast, Snb19 glioblastoma, and PC-3 prostate cancer cells to invade through matrigel. Invasion of PC-3 cells through matrigel was also restored by 34∶1 PC supplementation. Collectively, the data elucidate the novel metabolic regulation of invadopodia and the invasive process by de novo fatty acid synthesis and lipogenesis.