Articles de revues sur le sujet « Mathews, wilda »

These books offer ideas for dynamic professional development, theories about good teaching practices, ways to collaborate with colleagues, and important ideas and information about current issues in education. Reviewed are: Teamwork: Setting the Standard for Collaborative Teaching, Grades 5–9 by Monique Wild, Amanda Mayeaux, and Kathryn Edmonds; Teachers in Professional Communities: Improving Teaching and Learning by Ann Lieberman and Lynne Miller; Put Thinking to the Test by Lori Conrad, Missy Mathews, Cheryl Zimmerman, and Patrick Allen; and What Really Matters in Response to Intervention by Richard Allington.

Daffodils are an attractive group by their striking flowers and scents, therefore very appreciated as ornamental plants. Cultivars are very numerous, leading to more than 27,000 names considering them (Kington 2008). In the wild, the Iberian Peninsula can be recognized as the centre of speciation of Narcissus Linnaeus (1753: 289) (Barra et al. 2011). Consequently, the richest global diversity is found there (Hanks 2002). It is a complex genus that according to diverse authors varies from 25 to 87 species (cf. Webb 1980, Blanchard 1990, Mathew 2002, Zonneveld 2008, Aedo 2013, RHS 2016). Moreover, new species are still described (Escobar García 2018). Some daffodils have been described considering only herbarium vouchers. Nonetheless, it is encouraged to study populations from living specimens by two main reasons: (i) some features such as corona or perigone size—and ratios that include them—cannot be retrieved from exsiccata; and (ii) to study daffodils in the wild allow better understanding of their ecology and phenology (Barra Lázaro et al. 2016). The latter is important to clarify species spatial distribution and their hybrids.

Acute ozone (O3) inhalation has been shown to cause airway and pulmonary epithelial injury with accompanying inflammation responses. Robust evidence exists that O3 induces airway hyperresponsiveness (AHR) in humans and in animal models. Several pathways exist that culminate in airway smooth muscle contraction, but the mechanism(s) by which O3 elicits AHR are unclear. Here, we review the recent report by Kasahara et al. (Kasahara DI, Mathews JA, Park CY, Cho Y, Hunt G, Wurmbrand AP, Liao JK, Shore SA. Am J Physiol Lung Cell Mol Physiol 309: L736–L746, 2015.) describing the role of two Rho kinase (ROCK) isoforms in O3-induced AHR utilizing a murine haploinsufficiency model. Compared with wild-type (WT) mice, the authors report that ROCK1+/− and ROCK2+/− mice exhibited significantly reduced AHR following acute exposure to O3. Additionally, WT mice treated with fasudil, an FDA-approved ROCK1/2 inhibitor, recapitulated reduction in AHR as seen in ROCK haplotypes. It was suggested that, although the two ROCK isoforms are both induced by Rho, they have different mechanisms by which they mediate O3-induced AHR: ROCK1 via hyaluronan signaling vs. ROCK2 acting downstream of inflammation at the level of airway smooth muscle contraction. These observations provide an important framework to develop novel ROCK-targeting therapies for acute O3-induced AHR.

The plasma membrane calcium/calmodulin-dependent calcium ATPase (PMCA) (Shull, G.E., and J. Greeb. 1988. J. Biol. Chem. 263:8646–8657; Verma, A.K., A.G. Filoteo, D.R. Stanford, E.D. Wieben, J.T. Penniston, E.E. Strehler, R. Fischer, R. Heim, G. Vogel, S. Mathews, et al. 1988. J. Biol. Chem. 263:14152–14159; Carafoli, E. 1997. Basic Res. Cardiol. 92:59–61) has been proposed to be a regulator of calcium homeostasis and signal transduction networks of the cell. However, little is known about its precise mechanisms of action. Knock-out of (mainly neuronal) isoform 2 of the enzyme resulted in hearing loss and balance deficits due to severe inner ear defects, affecting formation and maintenance of otoconia (Kozel, P.J., R.A. Friedman, L.C. Erway, E.N. Yamoah, L.H. Liu, T. Riddle, J.J. Duffy, T. Doetschman, M.L. Miller, E.L. Cardell, and G.E. Shull. 1998. J. Biol. Chem. 273:18693–18696). Here we demonstrate that PMCA 4b is a negative regulator of nitric oxide synthase I (NOS-I, nNOS) in HEK293 embryonic kidney and neuro-2a neuroblastoma cell models. Binding of PMCA 4b to NOS-I was mediated by interaction of the COOH-terminal amino acids of PMCA 4b and the PDZ domain of NOS-I (PDZ: PSD 95/Dlg/ZO-1 protein domain). Increasing expression of wild-type PMCA 4b (but not PMCA mutants unable to bind PDZ domains or devoid of Ca2+-transporting activity) dramatically downregulated NO synthesis from wild-type NOS-I. A NOS-I mutant lacking the PDZ domain was not regulated by PMCA, demonstrating the specific nature of the PMCA–NOS-I interaction. Elucidation of PMCA as an interaction partner and major regulator of NOS-I provides evidence for a new dimension of integration between calcium and NO signaling pathways.

AbstractThe genusCrocusL. (Iridaceae) is monophyletic and contains about 100 species throughout the world.Crocusspecies have horticultural, medicinal and pharmacological importance. Saffron is the dried styles ofC. sativusand is one of the world’s most expensive spices by weight. Controversy exits about the taxonomy of the genus and the species relationship. Exploring genetic diversity and inter-specific cross-ability are important tasks for conservation of wild taxa and for breeding of cultivatedC. sativus. The present study was performed to study genetic variability and population structure in fiveCrocusL. species includingCrocus almehensisBrickell & Mathew,C. caspiusFischer & Meyer,C. speciosusMarschall von Biberstein,C. haussknechtiiBoissier, andC. sativusL. by inter simple sequence repeat (ISSR) molecular markers. We also used published internal transcribed spacer (ITS) sequences to study species relationship and compare the results with ISSR data. The results revealed a high degree of genetic variability both within and among the studied species. Neighbor joining (NJ) tree and network analysis revealed that ISSR markers are useful inCrocusspecies delimitation. Population fragmentation occurred inC. caspiusandC. sativus. Both ISSR and sequenced based analyses separatedC. sativusfrom the other studied species. Close genetic affinity ofC. sativusandC. pallisiiand inter-specific gene flow was supported by both data sets.

Abstract Knockdown of the growth hormone receptor (GHR) in melanoma cells downregulates ATP-binding cassette (ABC) transporters and sensitizes them to multiple drug treatments in vitro. The goal for this study is to understand whether a GHR antagonist (GHA) could suppress different types of cancers by sensitizing tumors to drug treatments in vivo through the downregulation of ABC transporters. Sera from GHA transgenic mice inhibited the proliferation of mouse melanoma cells (B16F10) in culture and suppressed expression of multiple ABC transporters. When B16-F10 cells were intradermally inoculated into GHA mice, tumor size was markedly reduced, as was STAT5 activation and ABCG1, ABCG2 levels. We then tested the effect of the GHA on the efficacy of cisplatin in vivo. GHA sensitized melanomas to cisplatin, leading to the smallest tumors among all groups. GHKO mice showed the same effect. We further extended this investigation to hepatocellular carcinoma (HCC). GHA mice were subcutaneously inoculated with mouse HCC (Hepa1 6 cells) and subsequently treated with sorafenib. The HCC tumors in GHA mice were markedly sensitive to sorafenib treatment compared to the same in wild-type mice. RNA analysis showed that when HCC was exposed to the combined treatment, relatively decreased ABC transporters expression was found. Immunohistochemical staining showed that phosphorylation of STAT5 and ABCB1 were downregulated in HCC tissues, suggesting that GHA in vivo downregulates ABC transporters in HCC, and therefore sensitizes them to sorafenib. Clinical data derived from HCC patients using the TCGA database showed that multiple ABC transporters in both types of cancer correlate with GHR levels; that is, when GHR levels are relatively high, patient survival is significantly decreased. Additionally, higher ABCC1 levels also lead to significantly decreased survival rate in HCC patients. Collectively, the results indicate that a GHA is effective in sensitizing both melanoma and HCC to available treatments in vivo and may be used as a therapeutic strategy for higher efficacy of tumor clearance. Citation Format: Yanrong Qian, Reetobrata Basu, Samuel Mathes, Prateek Kulkarni, Emily Davis, Darlene Berryman, Edward List, Silvana Duran, John J. Kopchick. Growth hormone receptor antagonist sensitizes melanoma and hepatocarcinoma to drug treatments via downregulation of ABC transporters in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2621.

Abstract Background: The diagnosis of intra-axial brain tumors requires histopathological examination of tissue obtained by neurosurgery in current clinical practice, which comes with inherent risks. Some patients, particularly those with primary CNS lymphoma (PCNSL), only undergo surgery to obtain a diagnosis and do not derive any therapeutic benefit from surgical resection or debulking. Less-invasive techniques for diagnosis, such as liquid biopsy, therefore provides an opportunity to mitigate surgical risk in these patients. Methods: Patients with histopathology confirmed glioblastoma (GBM, IDH wild type), brain metastases (BM), and PCNSL with accompanying cerebrospinal fluid (CSF) samples were included in our study. Cell-free DNA methylation profiling and shotgun proteomics were obtained for all patients and used to train classifiers to distinguish each tumour entity from others. Specifically, binomial elastic net regression models were built by combining both data modalities using established early and late integration paradigms and performance was compared to classifiers built from individual data types alone. Each model was repeated 100 fold and performance assessed on an untouched testing subset. Results: Our cohort includes 20 patients with GBM, 17 with BM, and 14 with PCNSL, each with matching CSF cell-free DNA methylation and shotgun proteomic profiling. We show that these data can be integrated to fully discriminate PCNSL from its major diagnostic counterparts with a perfect median AUC of 1.00 (95% CI 1-1) and 100% specificity. Integrated "lymphoma vs other" models significantly outperform models trained on methylation or protein data alone, though the same dramatic improvement was not demonstrated in GBM or BM, suggesting synergistic biological information is specific to lymphoma. Conclusions: There is a critical need to diagnose patients with intra-axial tumours, particularly PCNSL, without relying on invasive and costly surgery. We present the most specific and accurate CNS lymphoma classifier to date by integrating the methylome and proteome of CSF. This has the potential for immediate clinical utility, eliminating the need for biopsy in an important subset of these patients. Citation Format: Alex P. Landry, Jeffrey A. Zuccato, Vikas Patil, Mathew Voisin, Justin Z. Wang, Yosef Ellenbogen, Chloe Gui, Andrew Ajisebutu, Farshad Nassiri, Gelareh Zadeh. Integration of cerebrospinal fluid methylome and proteome can obviate the need for biopsy in central nervous system lymphoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1022.

Abstract Approximately 8-10% of pancreatic cancers do not harbor mutations in KRAS. Understanding the unique molecular and clinical features of this subset of pancreatic cancer (PC) is important to guide patient stratification for clinical trials of molecularly targeted agents. To this end we investigated a cohort of 795 PC patients from Dana-Farber Cancer Institute who had undergone somatic genomic characterization with OncoPanel, a targeted next-generation sequencing panel with coverage of more than 400 cancer-associated genes. A total of 9.2% (73/795) of cases in our cohort were KRAS WT. The KRAS WT cohort was statistically enriched for MSI-H PC and acinar cell carcinomas (p = 0.0035, p < 0.0001 respectively). Actionable alterations in alternative MAPK drivers were identified in 44% (32/73) of KRAS WT cases. BRAF alterations accounted for 56% (18/32) of detected alternative MAPK drivers, the majority of which (72%) were Class II which exhibit dimer-dependent constitutive activity. Receptor Tyrosine Kinase (RTK) fusion events in BRAF, NTRK1, NRG1, NTRK3, ROS1, and FGFR2 accounted for 25% (8/32) of detected alternative MAPK drivers in KRAS WT tumors. BRAF in-frame deletions showed increased sensitivity to dual pan-RAF and MEK inhibition in organoid models and one patient with a ROS1 fusion received prolonged clinical benefit from targeted therapy. In addition to alternative MAPK drivers, mutations in GNAS (p = 0.0014) and ARID2 (p = 0.045) were significantly enriched in KRAS WT PC, whereas TP53 mutations were significantly less frequent in KRAS WT cases. Interestingly, although not statistically significant, rates of mutation in the other canonical tumor suppressor genes (CDKN2A, SMAD4) were also lower in KRAS WT PC. Clinically, KRAS Mutant (MUT) PC were associated with a decreased overall survival (OS) compared to the KRAS WT cohort [median OS 17.5 vs 24.0 months, HR 1.38, p = 0.036], however this relationship was no longer significant after accounting for other clinical factors. For patients with KRAS WT PC, those with SMAD4 alterations had a significantly decreased OS (HR 6.24, p < 0.001), whereas presence of TP53 or CDKN2A mutations had no significant impact. Lastly, we found that KRAS WT PC was associated with a younger age of onset. Interestingly, we noted that KRAS WT PC patients with a younger age of onset had tumors with few oncogenic alterations whereas no such association was seen in KRAS MUT patients. Validation of this finding in a separate dataset is required and is currently ongoing. In summary, our clinical and genomic characterization of KRAS WT PC identifies a high prevalence of alternative MAPK drivers that are amenable to targeted therapies. Our cohort also recapitulates the previously reported clinical characteristics of KRAS WT PC and identifies the presence of SMAD4 alterations as significantly associated with decreased overall survival in KRAS WT PC. Additional analysis from multiple sources will be critical to risk stratify these patients further and to validate age-related findings. Citation Format: Harshabad Singh, Rachel B. Keller, Kevin S. Kapner, Julien Dilly, Srivatsan Raghavan, Chen Yuan, Eizabeth Cohen, Michael Tolstorukov, Emma Hill, Elizabeth Andrews, Lauren K. Brais, Annacarolina Da Silva, Kimberly Perez, Douglas A. Rubinson, Benjamin L. Schlechter, Michael H. Rosenthal, Jason L. Hornick, Valentina Nardi, Yvonne Li, Hersh Gupta, Andrew Cherniack, Mathew L. Meyerson, James M. Cleary, Jonathan A. Nowak, Brian M. Wolpin, Andrew A. Aguirre. Clinical-genomic analysis of KRAS wild-type pancreatic cancer confirms alternative targetable drivers and provides insight for age and risk related clinical stratification [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr A001.

Abstract Abstract 518 Targeted dosing of busulfan (Bu) has been shown to improve outcome of allogeneic HSCT (aHSCT) in patients with leukemia. There is limited data on correlation of Bu PK with outcome in patients with thalassemia major (TM)) undergoing aHSCT. We have previously shown that first dose trough level of Bu (Cmin1) predicts graft rejection (Chandy et al, BMT 2005), and Bu Css is significantly lower in patients with hepatic veno-occlusive disease (HVOD) (Srivastava et al, Blood 2004). The aim of the present study was to evaluate the correlations of Bu PK with outcome of HSCT in a larger cohort of patients and to evaluate the pharmacogenetic basis for the differences. We retrospectively analyzed oral Bu PK after 1st and 13th doses of Bu in 255 patients out of the 291 thalassemic patients who underwent aHSCT from matched related donors between 1991 till February 2010 at our centre. All patients received busulfan (at a dose of 14 or 16 mg/kg/day or 600mg/m2/day) in combination with Cy (at a dose of 160mg/kg for those >15 years or 200mg/kg for all others) as part of the conditioning regimen. Bu levels were measured by HPLC as previously described (Poonkuzhali et al, 1999). We also analyzed GSTA1*1B, GSTM1 and GSTT1 deletion polymorphisms in these patients. Based on Lucarelli's risk stratification, 18/291 patients belonged to class I, 121/291 were class II and 151/291 were class III. The class III patients were further risk stratified into class 3 high risk and low risk based on age and liver size (high risk: age >7 years and liver size >5cm; and the rest as low risk; Mathews et al, 2007). None of the Bu PK parameters were significantly different between Lucarelli classes as well as between class III high and low risk patients. For the entire group, EFS was 77%, OS 81%, NRM (non rejection mortality)15% and graft rejection 8.6%. Class III patients had a significantly lower EFS (p=0.0007) and OS (p=0.0051) compared to class I and II. Bu Cmin1 (p=0.007), but not Bu Css1 was significantly lower in those who rejected their graft compared to those who did not. On quartile analysis, patients with Cmin1 <156ng/ml had 30% (18/60) rejection compared to 8% (14/169) rejection in patients with Bu Cmin1 >156ng/ml (RR= 9.8; p=0.0001). Those with Bu Css1 in the lowest quartile also had significant risk of rejection (14/57 with Css1 <490 vs. 18/169 with Css1 >490 ng/ml; RR= 3.8, p=0.027) but the correlation was not as strong as that with Cmin1. Upon multivariate analysis of all the variables that were significantly influencing aHSCT outcome in univariate analysis, only Lucarelli class III high risk (p=0.034), SGOT level (p=0.036) and Bu Cmin1 (p=0.0001) were significantly influencing graft rejection. In addition, GSTA1*1B homozygous variant genotype was significantly associated with higher Bu Cmin1 (p=0.008) and Css1 (p=0.009). Since Bu Cmin1 was significantly influenced by GSTA1*1B genotype, we compared the combined risk of Cmin1 <156ng/ml and GSTA1*1B wild type genotype. None of the 6 patients with Bu Cmin1 <156ng/ml and GSTA1*1B homozygous mutant genotype rejected their graft. The incidence of graft rejection in the patients with GSTA1*1B wild type (17/113; 15%) and heterozygous (15/106; 9.4%) was higher than among those with homozygous mutant (1/34; 2.9%), (p=0.059). This is the largest available data on PK of oral busulfan patients with a single genetic disorder. We conclude that Bu Cmin1 is a stronger predictor of graft rejection than Css1 and that it is impacted by GSTA1*1B polymorphism in patients with beta thalassemia major undergoing aHSCT. Disclosures: Krishnamoorthy: INSERM U763: Employment, Research Funding.

Abstract Chemotherapy is often rendered ineffective by the development of multiple drug resistance (MDR). Given time MDR occurs almost universally and once developed applies in a non-specific manner. The lack of MDR specificity means many drugs, including those a patient has no pervious exposure to, are affected which limits treatment options. The goal of this research is to show nuclear localized tissue factor pathway inhibitor 1 (TFPI1) plays a driving role in development of MDR, and that the Anaphase Promoting Complex (APC) is capable of inhibiting MDR by targeting TFPI1 for degradation. A coagulation regulator, TFPI1 normally inhibits the tissue factor pathway to prevent blood clot formation. However, TFPI1 is highly elevated in many types of MDR cancer where we believe it mediates resistance development. Previous work in our lab has shown plasmid overexpression of TFPI1 induces MDR in MCF7 breast cancer cells, in MDR cells TFPI1 does not localize to the cell surface but instead to the nucleus, and a putative TFPI1 nuclear localization sequence (NLS) was identified. To confirm the putative NLS is required for TFPI1’s localization to nucleus, and subsequent MDR development, NLS mutant or wild type TFPI1-GFP were overexpressed in MCF7 cells. Localization of TFPI1-GFP was observed using widefield and confocal microscopy. Wild type TFPI1-GFP localized to the nucleus while the NLS mutant TFPI1 did not. These results help to confirm the putative NLS and its importance to TFPI1s role in MDR development. The APC is a nuclear E3 ubiquitin ligase and highly conserved regulator of cell cycle progression. Defects in APC function are associated with reduced longevity and cancer. Other work in our lab focuses on the APC’s role in longevity using a yeast model. However, we believe the APC also plays a role in TFPI1’s promotion of MDR. Our observations suggest APC dysfunction exists within MDR cancer cells where APC substrates accumulate, as does TFPI1. We hypothesize that nuclear TFPI1 is an APC substrate, and that APC activation will result in TFPI1 degradation slowing MDR development. Our lab has isolated several small peptides we believe bind and activate the APC, and have used these peptides in conjugation with known APC activators such as Mad2 Inhibitor-1 (M2I-1) to show that APC activation is capable of reducing TFPI1 abundance in MCF7 cells treated with the chemotherapeutic Doxorubicin (Dox). These results suggest TFPI1 is an APC substrate. To summarize, we believe TFPI1 drives development of MDR cancer, for which TFPI1’s accumulation within the nucleus is important. We have shown TFPI1’s NLS is required for localization of the protein to the nucleus, and activation of the APC reduces nuclear accumulation of TFPI1. Moving forward our goal is to demonstrate the effect TFPI1 NLS knockout and APC activation have on the development of MDR in MCF7 cells undergoing drug selection with Dox. By exploring TFPI1’s role in MDR we may elucidate means to slow resistance development and extend chemotherapy effectiveness. Citation Format: Mathew Lubachowski, Troy Harkness. Does TFPI1 drive development of multiple drug resistant cancer and can the anaphase promoting complex prevent it [abstract]. In: Proceedings of the AACR Special Conference: Aging and Cancer; 2022 Nov 17-20; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_1):Abstract nr B006.

Abstract Glioblastoma patients have a poor prognosis with a low median survival rate mainly due to temozolomide (TMZ) resistance. NRF2 is an important transcript factor involved in chemotherapy resistance due to its ability to regulate genes related to the antioxidant response and to prevent cell death processes, such as ferroptosis. However, the relation between NRF2 and iron-dependent cell death is contradictory and poorly understood. This study aimed to analyze the role of NRF2 in ferroptosis modulation in glioblastoma cells. To this end, it was analyzed two human glioblastoma cell lines (U251MG and T98G) after treatment with TMZ, ferroptosis inducers (Erastin, RSL3, and Sorafenib), and ferroptosis inhibitor (Ferrostatin-1). Also, we performed gene expression analysis of glioma patients. Our results demonstrated that T98G compared to the U251MG was more resistant to chemotherapy and showed elevated levels of NRF2 expression and its targets xCT, HMOX1, and ABCC1. Interestingly, T98G cells revealed higher sensitivity and lipoperoxidation levels after ferroptotic treatment. Next, we established T98G NRF2 silenced cells and we observed a significant reduction in cellular viability after TMZ treatment when compared to wild-type cells. On the other hand, T98G-shNRF2 was more resistant to ferroptosis induction, indicating that NRF2 plays a key role in the modulation of chemoresistance and ferroptosis. After, we showed that NRF2 controls levels of ABCC1/MRP1 in glioblastoma cells, and ABCC1 silencing promotes sensitivity to TMZ and resistance to Erastin. These results support a possible mechanism of ferroptosis modulation by NRF2 on TMZ-resistant gliomas through ABCC1, which has been recently associated with ferroptosis induction by promoting efflux of glutathione out of the cell. Furthermore, we confirmed that NRF2 has a positive correlation with ABCC1 in glioma patients, and higher ABCC1 expression was associated with tumor aggressiveness. Also, we validated ABCC1 as an independent prognostic factor for poor overall survival on glioma. Finally, we observed that T98G cells have sensitivity to the ferroptosis inducer FDA-approved, sorafenib. Altogether our data suggest that high levels of NRF2 may result in ferroptosis sensitivity on glioblastoma through the high levels of expression of its pro-ferroptotic target ABCC1, once the xCT system is blocked by Erastin. Thus, glioblastoma cell vulnerability to ferroptosis by NRF2 and ABCC1 high expression can be an Achilles’ heel to reverse drug resistance on glioblastoma through the treatment with ferroptosis inducers. Citation Format: Izadora de Souza, Linda Karolynne Monteiro, Camila Banca Guedes, Marcela Latancia, Marina Tomaz Andrade, Matheus Molina Silva, Bruna Contieri, Bruna Felício Milazzotto Ribeiro, Mariana Lazarini, Luciana Gomes, Clarissa Rocha. Temozolomide-resistant glioblastoma cells are collaterally sensitive to ferroptosis through NRF2 high expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5360.

Abstract Endocrine therapy forms the backbone treatment for patients with estrogen receptor (ER) positive tumors in both the adjuvant and metastatic setting. Aromatase inhibitors (AI) are the most common endocrine treatment option. Mutation of ESR1, the gene encoding ERα, is a common mechanism of resistance to AIs which leads to ligand independent activity of ERα. Camizestrant (AZD9833) is a next generation SERD and pure ER antagonist that is in Phase 3 trials (SERENA-4: NCT04711252; SERENA-6: NCT04964934). Here we report the preclinical and clinical activity of camizestrant in patients with ESR1 wild-type (ESR1wt) and mutant (ESR1m) tumors. The binding affinities of camizestrant, fulvestrant, and estradiol to wt ERα and ERα variants with mutations in the ligand binding domain were assessed. All three compounds exhibited reduced binding to mutant forms of ERα compared with wt ERα; the Y537S mutation had the greatest impact on binding. This was reflected in requirement for greater concentrations of camizestrant and fulvestrant to degrade and antagonize mutated ERα and to impact cellular proliferation in MCF-7 cells that expressed Y537S ESR1m compared to ESR1wt MCF-7 cells. Furthermore, while a 3 mg/kg dose of camizestrant achieved a maximal anti-tumor effect in a ESR1wt patient derived xenograft model, a 10 mg/kg was required for maximal effect in a D538G ESR1m model. Considering this difference between ESR1m and ESR1wt, pharmacokinetic/pharmacodynamic modelling of preclinical data predicted that a camizestrant dose of 75 mg would be maximally efficacious in patients with ESR1m tumors. Indeed, analysis of ESR1m circulating tumor DNA levels in patients from the SERENA-1 (NCT03616587) Phase 1 trial showed a clear effect of 14 days treatment with 75 mg camizestrant resulting in a >2-fold reduction in ESR1m variant allele frequency in 12/13 (92%) cases with complete clearance of ESR1m ctDNA in 7/13 (54%) cases. Interestingly, the clinical activity of camizestrant was higher in heavily pretreated patients with metastatic breast cancer with ESR1m tumors compared to those with no detectable mutation (ESR1m not detected). At a camizestrant dose of 75 mg, median progression-free survival was 8.3 months (maturity 12/15) in patients with ESR1m tumors compared to 5.6 months (8/9) in those with ESR1m not detected (data cut-off 6 October 2021). Camizestrant-induced ERα degradation was seen in both groups (mean reduction in H-score 42% in ESR1m tumors (n= 12 evaluable pairs) and 46% in tumors with ESR1m not detected (n=7)). Whole transcriptome analysis revealed a trend towards higher ERα activity at baseline in ESR1m tumors compared to ESR1m not detected; ERα activity reduced on treatment in both groups. Consistent with the clinical activity data, camizestrant induced more profound reductions in cell proliferation in ESR1m tumors compared to ESR1m not detected tumors (as seen by greater reductions in Ki67-positive tumor cells). These data demonstrate the activity of camizestrant in patients with ESR1m tumors. Clinical activity along with degradation and antagonism of the ERα is also seen in patients with tumors in which ESR1 mutations are not detected. In this heavily pre-treated Phase 1 patient population from SERENA-1, ESR1m may be a predictive biomarker to enrich for patients with maintained endocrine sensitivity. The SERENA-6 trial is investigating the efficacy and safety of camizestrant plus a CDK4/6 inhibitor in patients with metastatic breast cancer and detectable ESR1m. We acknowledge Helen Heffron, PhD, from InterComm International who provided medical writing support funded by AstraZeneca. Citation Format: Christopher Morrow, Larissa Carnevalli, Richard D. Baird, Tim Brier, Carmela Ciardullo, Natalie Cureton, Mandy Lawson, Robert McEwen, Myria Nikolaou, Anne Armstrong, Begoña Bermejo, Emiliano Calvo, Eva Ciruelos, Javier Garcia-Corbacho, Erika Hamilton, Jason Incorvati, Peter Kabos, Mafalda Oliveira, Manish R Patel, Manuel Ruiz-Borregó, Nicholas Turner, Chris Twelves, Christos Vaklavas, Danielle Carroll, Steven Ching, Nevena Cvetesic, Michelle DuPont, Lisa Gibbons, Alastair Mathewson, Rhiannon Maudsley, Pablo Morentin Gutierrez, Avinash Reddy, Jaime Rodriguez-Canales, Susana Ros, Dhivya Sudhan, Andy Sykes, David Whitson, Teresa Klinowska, Justin Lindemann. The next generation oral selective estrogen receptor degrader (SERD) camizestrant (AZD9833) is active against wild type and mutant estrogen receptor α [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-07-13.

Abstract The BRAF kinase plays a critical role in the MAPK signaling pathway and is mutated in ~8% of all human cancers including melanoma (~60%), thyroid (~60%), and lung adenocarcinoma (~10%). The most common mutation in BRAF is V600E (Class I), which is found in >70% in these cancers. Despite the clinical success of approved small molecule inhibitors of BRAF V600E (vemurafenib, dabrafenib and encorafenib), this remains an area of unmet medical need because nearly all patients progress, due to either primary or acquired resistance. A bifunctional degradation activating compound (or BiDACTM degrader) may address the liabilities of approved drugs by overcoming, or preventing the emergence of, resistance to BRAF inhibitors. Here we describe CFT1946, an orally bioavailable cereblon-based BiDAC degrader of BRAF V600 mutant proteins, and provide an overview of the medicinal chemistry path leading to its discovery. CFT1946 degrades BRAF V600 mutant proteins, while maintaining exquisite selectivity against the proteome, sparing wild type BRAF (BRAF-WT), ARAF, and CRAF. In A375 cells, CFT1946 potently degraded BRAF V600E and inhibited ERK phosphorylation and cell growth while having no effect in the mutant KRAS, BRAF-WT driven cell line HCT116. In the A375 xenograft model, oral delivery of CFT1946 at 10 mg/kg PO BID resulted in deeper and more durable tumor regression compared to a clinically relevant dose of encorafenib. Further evaluation of CFT1946 in an engineered, clinically relevant BRAFi-resistant A375 cell line (endogenous BRAF V600E + engineered expression of NRAS Q61K) demonstrated that CFT1946 both degraded BRAF V600E and caused a loss of viability in these cells, while treatment with encorafenib had no effect. In xenografts derived from this BRAFi-resistant cell line, oral dosing of CFT1946 as a single agent led to tumor growth inhibition, while treatment with a clinically relevant dose of encorafenib had no effect on tumor growth. Furthermore, dosing CFT1946 in combination with the MEK inhibitor, trametinib, resulted in significant tumor regression, whereas combining encorafenib with the same dose of trametinib had no effect. The medicinal chemistry campaign resulting in CFT1946 focused on the improvement of in vivo pharmacokinetics and rational linker design to achieve high oral bioavailability in a beyond Rule of 5 heterobifunctional degrader. The preclinical data presented herein support the planned Phase 1/2 clinical trial of CFT1946 for the treatment of BRAF-V600 mutant solid tumors. Citation Format: Yanke Liang, Mathew E. Sowa, Katrina L. Jackson, Jeffrey R. Simard, Bridget Kreger, Ping Li, Laura Poling, Joelle Baddour, Andrew Good, Hongwei Huang, Scott Eron, Christopher G. Nasveschuk, Robert Yu, Mark Fitzgerald, Victoria Garza, Morgan W. O’Shea, Gesine Veits, Jeremy Y. Yap, Moses Moustakim, Ashley Hart, Roman V. Agafonov, Grace Sarkissian, Joe S. Patel, Richard Deibler, Kyle S. Cole, David Cocozziello, Fazlur Rahman, Andrew J. Phillips, Elizabeth Norton, Adam S. Crystal, Roy M. Pollock, Stewart L. Fisher. The discovery and characterization of CFT1946: A potent, selective, and orally bioavailable degrader of mutant BRAF for the treatment of BRAF-driven cancers [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 3425.

Abstract While oncogenic KRAS is believed to drive pancreatic cancer by maintaining active ERK signaling, our studies suggest that ERK signaling can be also regulated in a KRAS-independent manner. We found that survival motor neuron domain containing 1 (SMNDC1), a poorly studied splicing factor is amplified or upregulated in 30% of pancreatic ductal adenocarcinomas (PDACs) and associated with lower survival rates in patients. To determine the role of SMNDC1 in PDACs, we conducted loss- and gain-of-function studies and found that this splicing factor drives tumor growth by promoting the retention of A/C-rich cassette exons, which otherwise are excluded or retained at a lower rate after RNA splicing in normal cells. Importantly, across tumors, SMNDC1 promoted the inclusion of the cassette exon 4 (E4) of MAPK3 (ERK1), which encodes the kinase-activating phosphorylation sites (Thr202/Tyr204). Forced exclusion of MAPK3-E4 using antisense oligonucleotide morpholino (ASO), inhibited tumor establishment, ERK1 phosphorylation, and expression of ERK-targets. We then tested if retention of E4 in MAPK3 by SMNDC1 is a mechanism to resist chemical inhibition of the KRAS-MEK-ERK pathway. We found that PDAC cells acutely treated with selective MEK inhibitors (MEKi), trametinib, or selumetinib increased SMNDC1 and MAPK3 E4 retention expression. In addition, PDAC cells deficient for SMNDC1 were 25-fold more sensitive to trametinib than isogenic cells expressing SMNDC1. To demonstrate that increased E4 retention drives MEKi resistance, we concomitantly treated SMNDC1 proficient PDAC cells with trametinib and E4 ASO, which led to an 8-fold increase in sensitivity compared to cells treated with trametinib and control ASO. Also, we found that resistance to standard-of-care chemotherapeutic agents to treat PDAC, Gemcitabine (GEM) and 5-fluorouracil (5-FU), was also associated with increased expression of SMNDC1 and retention of E4 in MAPK3 in multiple PDAC cells lines. To determine the biochemical mechanism that leads to E4 inclusion by SMNDC1, we performed high-throughput RNA Bind-n-Seq (RBNS) and e-CLIP of SMNDC1 in PDAC cells which revealed specific RNA-binding preference for C-rich polypyrimidine tract upstream of retained MAPK3-E4. To validate these findings, we constructed a dual-fluorescent reporter mini-gene of MAPK3-E4 splicing, with either wild-type sequences in E4 and polypyrimidine tract, or different point mutations at the predicted cis-elements. We found that retention of the E4 of MAPK3 is dependent on SMNDC1-binding to C-rich sequences at distant 3’ splice sites of PDAC cells. These data support that PDAC cells exploit a “splicing switch” to promote ERK kinase activity and resistance to therapies, independent of activating mutations in the EGFR/RAS/MAPK pathway and offer a druggable alternative to block oncogenic signaling and altered RNA splicing in PDAC. Citation Format: Md Afjalus Siraj, Yushan Zhang, Prabir Chakraborty, Robert Tseng, Li-Ting Ku, Grant Goda, Gilbert Giri, Deanne Yugawa, Mathew Wang, Shamik Das, Anindya Dey, Shailendra Dhar Dwivedi, Geeta Rao, Min Zhang, Da Yang, Md Nazir Hossen, Wei-Qun Ding, Kar-Ming Fung, Daniel Dominguez, Resham Bhattacharya, Luisa Escobar-Hoyos, Priyabrata Mukherjee. SMNDC1 alters the splicing of ERK to potentiate its activity in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr C083.

Abstract A toxicity reduced conditioning regimen containing Treosulfan (Treo), fludarabine (Flu), thiotepa for high risk Thal Major (TM) has been used since 2009 at our centre that has significantly improved transplant outcomes of these patients compared to the historical cohort of patients receiving busulfan/ cyclophosphamide based myeloablative regimen (Mathews et al, 2013). Limited knowledge is available on the pharmacokinetics (PK), pharmacogenetics (PG) and pharmacodynamics of fludarabine and treosulfan, especially in non-malignant hematological disorders like TM. We describe here the PK of Flu and Treo in patients with TM undergoing HSCT, the factors influencing the inter-individual variability in PK and the role of these factors on HSCT outcome. Seventy one patients diagnosed with TM undergoing HSCT with Flu/Treo based conditioning regimen between January 2012 and January 2015 were included (Table: Patient demographics). Selected functional polymorphisms in the NT5E, DCK, hENT1 and GST genes that are involved in fludarabine or treosulfan metabolism were screened. All patients received Flu 40mg/m2/day x 4 days as an 1hr infusion on days 1 and 4 and Treo as 14g/m2/day x 3 days at the rate 5g/hr. Plasma was separated from the peripheral blood collected at predetermined time points after the infusion of Flu and Treo PK analysis. Plasma Flu was analyzed using a LC-MS/MS method and the concentration was expressed as mMole/ml while Treo was analyzed using a HPLC-RI method and concentration was expressed as mg/L. Flu and Treo PK was estimated using nonlinear mixed effects modeling via Monolix 4.3.3. The covariates tested for both PK were: age, sex, body weight, BSA, ferritin, and polymorphisms in NT5E, hENT1, dCK and GST genes. The PK parameters AUC, CL, V and k were estimated on day 1 for Treo and on day 1 and day 4 for Flu (Table). The influence of Flu and Treo PK and PG on graft rejection, early transplant related mortality (TRM) & chimerism status was estimated using logistic regression analysis. Wide inter-individual variation in Flu and Treo PK was noted (7 and 9 fold Vs 5 and 8 fold respectively for Day 1 & 4 Flu AUC & Cl; 33 & 31 fold variation in Treo AUC and Cl) (Table). Flu CL was significantly higher on day 4 compared to day1 (Figure A). The variation in Flu PK was explained by genetic variants in NT5E and dCK. Patients having variant genotype for the SNPs in NT5E (rs2295890) and dCK (rs11544786) showed significantly lower plasma Flu clearance compared to those with wild type genotype (p=0.006 & p=0.05 respectively) (Figure B). This is consistent with our previous report in patients with aplastic anemia undergoing HSCT (Mohanan et al. 2014; Blood: 124 (21)). None of the genetic variants in the GST genes explained the variation in Treo PK. Day21 mortality was seen in 6/71 patients (8.5%) and graft rejection in 3/66 evaluable patients (4.5%). Analysis of the influence of PK and PG variables on transplant outcome showed significantly high first dose Flu AUC to be associated with D21 mortality upon Univariate analysis (median 42.5, range 32.1-63.7 compared to 31.8, range 15.2-111 mMole*h/mL, in those with and without TRM respectively; p=0.043); none of these parameters were significantly associated with graft rejection or mixed chimerism. There was no association between Treo PK parameters and graft rejection or TRM. The influence of Flu and Treo PK on regimen related toxicity is yet to be evaluated. The lack of the influence of PK on transplant outcome could be due to lower incidence of rejection and TRM in this cohort. Further analysis in a larger cohort of patients will be done once we enroll more patients for PK analysis. Our results demonstrate that Flu PK is influenced by genetic variants in NT5E and dCK, the enzymes involved in Flu biotransformation. The relationship between high-plasma Flu exposure and TRM and given the fact that multiple factors influence TRM, we can extrapolate that the plasma Flu AUC may be a surrogate marker of overall preparative regimen intensity as reported previously (Long-Boyle et al, Bone Marrow Transplant, 2011). The lack of association of genetic variants in GST genes in explaining the inter-patient variability in treosulfan exposure suggests the involvement of other drug metabolizing genes on treosulfan PK. We are currently evaluating the role of genetic variants in a large panel of drug metabolizing genes on explaining this inter-individual variability in Treo PK. Disclosures No relevant conflicts of interest to declare.

Abstract Introduction: Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin's lymphoma (NHL), accounting for roughly 30% of newly diagnosed cases in the US. DLBCL can progress quickly, and in advanced cases is inconsistently cured with current therapies. Ibrutinib, a first-in-class Bruton's tyrosine kinase (BTK) inhibitor, is approved as a treatment for patients (pts) with mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL) who have had one prior treatment. The ABC subtype of DLBCL is considered especially high risk and characterized by chronic active B-cell receptor (BCR) signaling, which is blocked by ibrutinib. Recent phase II clinical trial results of ibrutinib as a single agent in DLBCL pts show an overall response rate of 41% in the ABC subtype (Wilson et al. ASH 2012). Responses of various cancers to single kinase targeted therapies are often limited by the cell's ability to bypass the target via alternative pathways or acquired mutations in the target or its pathway. It has been shown that a small number of CLL pts acquire resistance to ibrutinib through mutations in BTK and its substrate phospholipase C gamma 2 (PLCG2) in the B lymphoma cells following prolonged treatments (Woyach et al. NEJM 2014). Such mechanisms may be overcome by combinations of targeted agents. Through screening of wild-type and acquired ibrutinib-resistant ABC-DLBCL cell lines (e.g. expressing BTK C481S), we identify and report herein B-cell lymphoma-2 (BCL-2) and spleen tyrosine kinase (SYK) inhibitors that synergize with ibrutinib in vitro and in vivo. Methods: Gene expression was analyzed by RT-qPCR using TaqMan Gene Expression Master Mix. Human DLBCL cell lines were treated with drugs for 3 days and the effect on cell growth was determined by CellTiter-Glo luminescent cell viability assay. SCID mice were treated when the TMD8 tumors reached 100-150 mm3. Annexin-V-positive and PI-negative population was detected as apoptotic cells in tumor cells at sacrifice. Cell adhesion and migration assays were performed as previously described (Chang et al. Blood 2013). Analysis of clinical samples used for BCL-2 gene expression profiling was performed using Affymetrix microarrays on FFPE specimens from the phase 2 PCYC-1106 trial (NCT01325701) and a rank based statistic (RankProd) was used to determine the significance of gene expression changes. Results: DLBCL cell lines with higher BCL-2 expression were more sensitive to single agent ABT-199 than those with lower expression. Treatment of DLBCL cells with ibrutinib alone increased BCL-2 expression as well as their sensitivity to BCL-2 inhibitors. Combination treatment with BCL-2 inhibitors and ibrutinib completely inhibited tumor growth in murine models of ABC-DLBCL (Figure). Increased apoptotic cell populations were detected in the combination treated tumors compared to either treatment alone. Clinically, pretreatment tissue samples (n=28) from ABC-DLBCL pts who experienced objective responses to ibrutinib (CR+PR) had lower BCL-2 gene expression. A high BCL-2 mutation rate was observed in pts with poor response to ibrutinib (SD+PD). However, none of these mutations occurred in the BH3 or BH1 domains, both of which appear to interact with ABT-199 based on a 3-dimensional co-crystal structure of the inhibitor with BCL-2 (PDB code 4MAN) and further molecular simulation results. These findings suggest the potential benefit from combination therapy. SYK is another downstream mediator of BCR signaling. Pretreatment of DLBCL cells with SYK inhibitors (e.g. R406) increased their sensitivity to ibrutinib. Ibrutinib resistant B-lymphoma cells with either C481S BTK or R665W PLCG2 mutations were re-sensitized to ibrutinib in combination with BCL-2 or SYK inhibitors, inhibiting cell growth, IgM-induced calcium flux, cell adhesion or migration in mutant containing cells. Conclusions: Consistent with previous results from high-throughput combinatorial screenings of drugs interact favorably with ibrutinib (Mathews Griner, et al. PNAS 2013), we found BCL-2 and SYK may function in alternative survival pathways in DLBCL cells upon BTK inhibition. Human B lymphomas harboring ibrutinib-resistant C481S BTK or R665W PLCG2 may be re-sensitized by BCL-2 or SYK inhibitors, both of which provide a rationale for the design of combination therapies. Figure 1 Combination of ibrutinib and ABT-199 on the effect of TMD-8 tumor growth. Figure 1. Combination of ibrutinib and ABT-199 on the effect of TMD-8 tumor growth. Disclosures Kuo: Pharmacyclics: Employment. Crowley:Pharmacyclics: Employment. Xue:Pharmacyclics: Employment. Schweighofer:Pharmacyclics: Employment. Cheung:Pharmacyclics: Employment. Hsieh:Pharmacyclics: Employment. Eckert:Pharmacyclics: Employment. Versele:Janssen Pharmaceutica: Employment. Chang:Pharmacyclics, Inc: Employment, Equity Ownership.

Rose rosette disease is an incurable, destructive disease that affects both wild and cultivated roses. Over the past several decades, the disease has spread over much of the U.S., though it was first observed in Florida in 2013. This 6-page fact sheet describes the symptoms and diagnosis of the disease, as well as the cultural, chemical, and, possibly, biological controls that can minimize its spread. Written by Binoy Babu, Mathews L. Paret, Tim Schubert, Carlye Baker, Gary Knox, Fanny Iriarte, James Aldrich, Laura Ritchie, Carrie L. Harmon, and Svetlana Y. Folimonova, and published by the UF Department of Plant Pathology, May 2015. http://edis.ifas.ufl.edu/pp317

This research primarily deals with mathematically ranking the level of accuracy of various Artificial Intelligence (AI) based machine learning classifiers, using Mathews Correlation Coefficient (MCC), leveraging Confusion Matrices. A detailed Literature survey was done to gather the existing knowledge. This knowledge was used as foundational basis to further build the scope of this research project. The classifiers used were Support Vector Machine (SVM), K-Nearest Neighbors (KNN), and Convolutional Neural Network (CNN). These classifiers were used for Face recognition of individuals. A total of 33 adult test subjects with 10 images per subject resulting in a total of 330 distinct images were part of this research project. These test subjects were from the Labeled Faces in The Wild publicly available database along with teachers from James B. Conant High School who voluntarily participated in this research. Python based face recognition program wrappers and associated environment was built around pre-existing classifiers and the image data was passed to these wrappers. These wrappers recognized the faces of individuals in the images over 10 trials wherein each trial consisted of 33 distinct images, with varying degree of accuracy and presented that as outputs. These outputs were used to determine the Confusion Matrices which in turn were used to calculate the MCC scores. The MCC scores were plotted, and these results showed that the SVM AI classifier had the highest level of relative accuracy for face recognition, followed by KNN and CNN classifiers.

The role of literature in the formation of culture is an oft repeated and much discussed topic of interest in any civilized world. In spite of that, very seldom we realize the sanctity and validity of this discourse. It is a grim reality that the modern world looks at odds with its cultural concerns and subsequently it alienates itself from the framework of responsibility, which is an utter disregard for a developing culture. Being a preliminary branch of art, literature serves a dual role in the scheme of things for the cultural development. On the other hand, it is an outcome of cultural progression, because the literary treasure of any social group is a part of its culture. On the other hand literature provides all the necessary ingredients for the formation of a culture – complex, that is an aggregate of cultural ethos. Mathew Arnold described art as a “criticism of life”. This view was an extension of Aristotle’s belief that art depends upon the imitation of nature. There was however some other scholars like Oscar Wilde, who believed that life itself imitates art. Both these theories are interesting and vital in understanding the concerns of literature as a work of art in the construction and preservation of culture. At the same time we have to formulate a canon for delineating the cultural concerns of literature. The topic of this research paper is an attempt to draw our attention toward this fundamental issue, that relates the human society to a larger canvass of its designated ideals.

CONTENTS: FOREWORD PAUL LAURENCE DUNBAR • Ere Sleep Comes Down to Soothe the Weary Eyes • Death Song • Life • After the Quarrel • Ships that Pass in the Night • We Wear the Mask • Sympathy • The Debt JOSEPH S. COTTER, SR • The Tragedy of Pete • The Way-side Well JAMES WELDON JOHNSON • From the German of Uhland • The Glory of the Day Was in Her Face • The Creation • The White Witch • My City WILLIAM EDWARD BURGHARDT Du BOIS • A Litany of Atlanta WILLIAM STANLEY BRAITHWAITE • Scintilla • Rye Bread • October XXIX, 1795 • Del Cascar JAMES EDWARD MCCALL • The New Negro ANGELINA WELD GRIMKE • Hushed by the Hands of Sleep • Greenness • • The Eyes of My Regret • Grass Fingers • Surrender • The Ways o' Men • Tenebris • When the Green Lies Over the Earth • A Mona Lisa • Paradox • Your Hands • I Weep • For the Candle Light • Dusk. • The Puppet Player • A Winter Twilight ANNE SPENCER • Neighbors • I Have a Friend • Substitution • Questing • Life-long, Poor Browning • Dunbar • Innocence • Creed • Lines to a Nasturtium • At the Carnival MARY EFFIE LEE NEWSOME • Morning Light • Pansy • Sassafras Tea • Sky Pictures • The Quilt • The Baker's Boy • Wild Roses • Quoits JOHN FREDERICK MATHEUS • Requiem FENTON JOHNSON • When I Die • Puck Goes to Court • The Marathon Runner • JESSIE FAUSET • Words! Words! • Touche • Noblesse Oblige • La Vie C'est la Vie • The Return • Rencontre • Fragment ALICE DUNBAR NELSON • Snow in October • Sonnet • I Sit and Sew GEORGIA DOUGLAS JOHNSON • Service • Hope • The Suppliant • Little Son • Old Black Men • Lethe • Proving • I Want to Die While You Love Me • Recessional • My Little Dreams • What Need Have I for Memory? • When I Am Dead • The Dreams of the Dreamer • The Heart of a Woman CLAUDE McKAy • America • Exhortation: Summer, 1919 • Flame-heart • The Wild Goat • Russian Cathedral • Desolate • Absence • My House JEAN TOOMER • Reapers • Evening Song • Georgia Dusk • Song of the Son • Cotton Song • Face • November Cotton Flower JOSEPH S. COTTER, JR • Rain Music • Supplication • An April Day • The Deserter • And What Shall You Say? • The Band of Gideon BLANCHE TAYLOR DICKINSON • The Walls of Jericho • Poem • Revelation • That Hill • To an Icicle • Four Walls FRANK HORNE • On Seeing Two Brown Boys in a Catholic Church • To a Persistent Phantom • Letters Found Near a Suicide • Nigger LEWIS ALEXANDER • Negro Woman • Africa • Transformation • The Dark Brother • Tanka I-VIII • Japanese Hokku • Day and Night STERLING A. BROWN • Odyssey of Big Boy • Maumee Ruth • Long Gone • To a Certain Lady, in Her Garden • Salutamus • Challenge • Return CLARISSA SCOTT DELANY • Joy • Solace • Interim • The Mask LANGSTON HUGHES • I, Too • Prayer • Song for a Dark Girl • Homesick Blues • Fantasy in Purple • Dream Variation • The Negro Speaks of Rivers • Poem • Suicide's Note • Mother to Son • A House in Taos GWENDOLYN B. BENNETT • Quatrains • Secret • Advice • To a Dark Girl • Your Songs • Fantasy • Lines Written at the Grave of Alexander Dumas • Hatred • Sonnet—l • Sonnet—2 AnNA BONTEMPS • The Return • A Black Man Talks of Reaping • To a Young Girl Leaving the Hill Country • Nocturne at Bethesda • Length of Moon • Lancelot • Gethsemane • A Tree Design • Blight • The Day-breakers • Close Your Eyes! • God Give to Men • Homing • Golgotha Is a Mountain ALBERT RICE • The Black Madonna • To a Certain Woman COUNTEE CULLEN • I Have a Rendezvous with Life • Protest • Yet Do I Marvel • To Lovers of Earth: Fair Warning • From the Dark Tower • To John Keats, Poet, at Springtime • Four Epitaphs • Incident DONALD JEFFREY HAYES • Inscription • Auf Wiedersehen • Night • Confession • Nocturne • After All • JONATHAN HENDERSON BROOKS • The Resurrection • The Last Quarter Moon of the Dying Year • Paean GLADYS MAY CASELY HAYFORD • Nativity • Rainy Season Love Song • The Serving Girl • Baby Cobina LuCY ARIEL WILLIAMS • Northboun' GEORGE LEONARD ALLEN • To Melody • Portrait RICHARD BRUCE • Shadow • Cavalier WARING CUNEY • The Death Bed • A Triviality • I Think I See Him There • Dust • No Images • The Radical • True Love EDWARD S. SILVERA • South Street • Jungle Taste HELENE JOHNSON • What Do I Care for Morning • Sonnet to a Negro in Harlem • Summer Matures • Poem • Fulfillment • The Road • Bottled • Magalu WESLEY CURTWRIGHT • The Close of Day LULA LOWE WEEDEN • Me Alone • Have You Seen It • Robin Red Breast 228 • The Stream • The Little Dandelion • Dance

IntroductionI work at a regional campus of La Trobe University, Australia. More precisely, I work at the Bendigo campus of La Trobe University. At Bendigo, we are often annoyed when referred to and addressed as ‘regional’ students and staff. Really, we should not be. After all, Bendigo campus is an outpost of La Trobe’s metropolitan base. It is funded, run, and directed from Bundoora (Melbourne). The word ‘regional’ simply describes the situation. A region is an “administrative division of a city or a district [… or …] a country” (Brown 2528). And the Latin etymology of region (regio, regere) includes “direction, line”, and “rule” (Kidd 208, 589). Just as the Bendigo campus of La Trobe is a satellite of the metropolitan campus, the town of Bendigo is an outpost of Melbourne. So, when we are addressed and interpellated (Althusser 48) as regional, it is a reminder of the ongoing fact that Australia is (still) a colony, an outpost of empire, a country organised on the colonial model. From central administrative hubs, spokes of communication, and transportation spread to the outposts. When Bendigo students and staff are addressed as regional, in a way we are also being addressed as colonial.In this article, the terms ‘region’ and ‘regional’ are deployed as inextricably associated with the Australian version of colonialism. In Australia, in the central metropolitan hubs, where the colonial project is at its most comprehensive, it is hard to see what remains, to see what has escaped that project. The aim of this article is to explore how different aspects of the country escape the totalising project of Australian colonialism. This exploration is undertaken primarily through a discussion of the ways in which some places on this continent remain not regional (and thus, not colonial) how they keep the metropolis at bay, and how they, thus, keep Europe at bay. This discussion includes a general overview of the Australian colonial project, particularly as it pertains to First Nations Peoples, their knowledge and philosophies, and the continent’s unique ecologies. Then the article becomes more speculative, imagining different ways of seeing and experiencing time and place in this country, ways of seeing the remains and refuges of pre-1788, not-regional, and not-colonial Australia. In these remains and refuges, there persist the flourishing and radical difference of this continent’s ecologies and, not surprisingly, the radical suitedness of tens of thousands of years of First Nations Peoples’ culture and thinking to that ecology, as Country. In what remains not regional, I argue, are answers to the question: How will we live here in the Anthropocene?A Totalising ProjectSince 1788, in the face of the ongoing presence and resistance of First Nations cultures, and the continent’s radically unique ecologies, the Australian colonial project has been to convert the continent into a region of Europe. As such, the imposed political, administrative, scientific, and economic institutions are largely European. This is also so, to a lesser extent, of social and cultural institutions. While the continent is not Europe geologically, the notion of the Anthropocene suggests that this is changing (Crutzen and Stoermer). This article does not resummarise the vast body of scholarship on the effects of colonisation, from genocide to missionary charity, to the creation of bureaucratic and comprador classes, and so on. Suffice to say that the different valences of colonisation—from outright malevolence to misguided benevolence–produce similar and common effects. As such, what we experience in metropolitan and regional Australia, is chillingly similar to what people experience in London. Chilling, because this experience demonstrates how the effects of the project tend towards the total.To clarify, when I use the name ‘Australia’ I understand it as the continent’s European name. When I use the term ‘Europe’ or ‘European’, I refer to both the European continent and to the reach and scope of the various colonial and imperial projects of European nations. I take this approach because I think it is necessary to recognise their global effects and loads. In Australia, this load has been evident and present for more than two centuries. On one hand, it is evident in the social, cultural, and political institutions that come with colonisation. On another, it is evident in the environmental impacts of colonisation: impacts that are severely compounded in Australia. In relation to this, there is vital, ongoing scholarship that explores the fact that, ecologically, Australia is a radically different place, and which discusses the ways in which European scientific, aesthetic, and agricultural assumptions, and the associated naturalised and generic understandings of ‘nature’, have grounded activities that have radically transformed the continent’s biosphere. To name but a few, Tim Flannery (Eaters, “Ecosystems”) and Stephen Pyne, respectively, examine the radical difference of this continent’s ecology, geology, climate, and fire regimes. Sylvia Hallam, Bill Gammage, and Bruce Pascoe (“Bolt”, Emu) explore the relationships of First Nations Peoples with that ecology, climate, and fire before 1788, and the European blindness to the complexity of these relationships. For instance, William Lines quotes the strikingly contradictory observations of the colonial surveyor, Thomas Mitchell, where the land is simultaneously “populous” and “without inhabitants” and “ready for the immediate reception of civilised man” and European pastoralism (Mitchell qtd. in Lines 71). Flannery (Eaters) and Tim Low (Feral, New) discuss the impacts of introduced agricultural practices, exotic animals, and plants. Tom Griffiths tells the story of ‘Improving’ and ‘Acclimatisation Societies’, whose explicit aims were to convert Australian lands into European lands (32–48). The notion of ‘keeping Europe at bay’ is a response to the colonial assumptions, practices, and impositions highlighted by these writers.The project of converting this continent and hundreds of First Nations Countries into a region of Europe, ‘Australia’, is, in ambition, a totalising one. From the strange flag-plantings, invocations and incantations claiming ownership and dominion, to legalistic conceptions such as terra nullius, the aim has been to speak, to declare, to interpellate the country as European. What is not European, must be made European. What cannot be made European is either (un)seen in a way which diminishes or denies its existence, or must be made not to exist. These are difficult things to do: to not see, to unsee, or to eradicate.One of the first acts of administrative division (direction and rule) in the Port Phillip colony (now known as Victoria) was that of designating four regional Aboriginal Protectorates. Edward Stone Parker was appointed Assistant Protector of Aborigines for the Loddon District, a district which persists today for many state and local government instrumentalities as the Loddon-Mallee region. In the 1840s, Parker experienced the difficulty described above, in attempting to ‘make European’ the Dja Dja Wurrung people. As part of Parker’s goal of Christianising Dja Dja Wurrung people, he sought to learn their language. Bain Attwood records his frustration:[Parker] remarked in July 1842. ‘For physical objects and their attributes, the language readily supplies equivalent terms, but for the metaphysical, so far I have been able to discover scarcely any’. A few years later Parker simply despaired that this work of translation could be undertaken. ‘What can be done’, he complained, ‘with a people whose language knows no such terms as holiness, justice, righteousness, sin, guilt, repentance, redemption, pardon, peace, and c., and to whose minds the ideas conveyed by those words are utterly foreign and inexplicable?’ (Attwood 125)The assumption here is that values and concepts that are ‘untranslatable’ into European understandings mark an absence of such value and concept. Such assumptions are evident in attempts to convince, cajole, or coerce First Nations Peoples into abandoning traditional cultural and custodial relationships with Country in favour of individual private property ownership. The desire to maintain relationships with Country are described by conservative political figures such as Tony Abbott as “lifestyle choices” (Medhora), effectively declaring them non-existent. In addition, processes designed to recognise First Nations relationships to Country are procedurally frustrated. Examples of this are the bizarre decisions made in 2018 and 2019 by Nigel Scullion, the then Indigenous Affairs Minister, to fund objections to land claims from funds designated to alleviate Indigenous disadvantage and to refuse to grant land rights claims even when procedural obstacles have been cleared (Allam). In Australia, given that First Nations social, cultural, and political life is seamlessly interwoven with the environment, ecology, the land–Country, and that the colonial project has always been, and still is, a totalising one, it is a project which aims to sever the connections to place of First Nations Peoples. Concomitantly, when the connections cannot be severed, the people must be either converted, dismissed, or erased.This project, no matter how brutal and relentless, however, has not achieved totality.What Remains Not Regional? If colonisation is a totalising project, and regional Australia stands as evidence of this project’s ongoing push, then what remains not regional, or untouched by the colonial? What escapes the administrative, the institutional, the ecological, the incantatory, and the interpellative reach of the regional? I think that despite this reach, there are such remains. The frustration, the anger, and antipathy of Parker, Abbott, and Scullion bear this out. Their project is unfinished and the resistance to it infuriates. I think that, in Australia, the different ways in which pre-1788 modes of life persist are modes of life which can be said to be ‘keeping Europe at bay’.In Reports from a Wild Country: Ethics for Decolonisation, Deborah Bird Rose compares Western/European conceptualisations of time, with those of the people living in the communities around the Victoria River in the Northern Territory. Rose describes Western constructions of time as characterised by disjunction (for example, the ‘birth’ of philosophy, the beginnings of Christianity) and by irreversible sequence (for example, concepts of telos, apocalypse, and progress). These constructions have become so naturalised as to carry a “seemingly commonsensical orientation toward the future” (15). Orientation, in an Australian society “built on destruction, enables regimes of violence to continue their work while claiming the moral ground of making a better future” (15). Such an orientation “enables us to turn our backs on the current social facts of pain, damage, destruction and despair which exist in the present, but which we will only acknowledge as our past” (17).In contrast to this ‘future vision’, Rose describes what she calls the ‘canonical’ time-space conceptualisation of the Victoria River people (55). Here, rather than a temporal extension into an empty future, orientation is towards living, peopled, and grounded origins, with the emphasis on the plural, rather than a single point of origin or disjunction:We here now, meaning we here in a shared present, are distinct from the people of the early days by the fact that they preceded us and made our lives possible. We are the ‘behind mob’—those who come after. The future is the domain of those who come after us. They are referred to as […] those ‘behind us’. (55)By way of illustration, when we walk into a sheep paddock, even if we are going somewhere (even the future), we are also irrevocably walking behind ancestors, predecessor ecologies, previous effects. The paddock, is how it is, after about 65,000 years of occupation, custodianship, and management, after European surveyors, squatters, frontier conflict and violence, the radical transformation of the country, the destruction of the systems that came before. Everything there, as Freya Mathews would put it, is of “the given” (“Becoming” 254, “Old” 127). We are coming up behind. That paddock is the past and present, and what happens next is irrevocably shaped by it. We cannot walk away from it.What remains not regional is there in front of us. Country, language, and knowledge remain in the sheep paddock, coexisting with everyone and everything else that everyone in this country follows (including the colonial and the regional). It is not gone. We have to learn how to see it.By the Fox or the Little EagleFigure 1: A Scatter of Sulphur-Crested Cockatoo Feathers at Wehla. Image Credit: Terry Eyssens.As a way of elaborating on this, I will tell you about a small, eight hectare, patch of land in Dja Dja Wurrung Country. Depending on the day, or the season, or your reason, it could take fifteen minutes to walk from one end to the other or it might take four hours, from the time you start walking, to the time when you get back to where you started. At this place, I found a scatter of White Cockatoo feathers (Sulphur-Crested Cockatoo—Cacatua galerita). There was no body, just the feathers, but it was clear that the Cockatoo had died, had been caught by something, for food. The scatter was beautiful. The feathers, their sulphur highlights, were lying on yellow-brown, creamy, dry grass. I dwelled on the scatter. I looked. I looked around. I walked around. I scanned the horizon and squinted at the sky. And I wondered, what happened.This small patch of land in Dja Dja Wurrung Country is in an area now known as Wehla. In the Dja Dja Wurrung and many other Victorian languages, ‘Wehla’ (and variants of this word) is a name for the Brushtail Possum (Trichosurus vulpecula). In the time I spend there/here, I see all kinds of animals. Of these, two are particularly involved in this story. One is the Fox (Vulpes vulpes), which I usually see just the back of, going away. They are never surprised. They know, or seem to know, where everyone is. They have a trot, a purposeful, cocky trot, whether they are going away because of me or whether they are going somewhere for their own good reasons. Another animal I see often is the Little Eagle (Hieraaetus morphnoides). It is a half to two-thirds the size of a Wedge-tailed Eagle (Aquila audax). It soars impressively. Sometimes I mistake a Little Eagle for a Wedge-tail, until I get a better look and realise that it is not quite that big. I am not sure where the Little Eagle’s nest is but it must be close by.I wondered about this scatter of White Cockatoo feathers. I wondered, was the scatter of White Cockatoo feathers by the Fox or by the Little Eagle? This could be just a cute thought experiment. But I think the question matters because it provokes thinking about what is regional and what remains not regional. The Fox is absolutely imperial. It is introduced and widespread. Low describes it as among Australia’s “greatest agent[s] of extinction” (124). It is part of the colonisation of this place, down to this small patch of land in Dja Dja Wurrung Country. Where the Fox is, colonisation, and everything that goes with it, remains, and maintains. So, that scatter of feathers could be a colonial, regional happening. Or maybe it is something that remains not regional, not colonial. Maybe the scatter is something that escapes the regional. The Little Eagles and the Cockatoos, who were here before colonisation, and their dance (a dance of death for the Cockatoo, a dance of life for the Little Eagle), is maybe something that remains not regional.But, so what if the scatter of White Cockatoo feathers, this few square metres of wind-blown matter, is not regional? Well, if it is ‘not regional’, then, if Australia is to become something other than a colony, we have to look for these things that are not regional, that are not colonial, that are not imperial. Maybe if we start with a scatter of White Cockatoo feathers that was by the Little Eagle, and then build outwards again, we might start to notice more things that are not regional, that still somehow escape. For example, the persistence of First Nations modes of land custodianship and First Nations understandings of time. Then, taking care not to fetishise First Nations philosophies and cultures, take the time and care to recognise the associations of all of those things with simply, the places themselves, like a patch of land in Dja Dja Wurrung Country, which is now known as Wehla. Instead of understanding that place as something that is just part of the former Aboriginal Protectorate of Loddon or of the Loddon Mallee region of Victoria, it is Wehla.The beginning of decolonisation is deregionalisation. Every time we recognise the not regional (which is hopefully, eventually, articulated in a more positive sense than ‘not regional’), and just say something like ‘Wehla’, we can start to keep Europe at bay. Europe’s done enough.seeing and SeeingChina Miéville’s The City and The City (2009) is set in a place, in which the citizens of two cities live. The cities, Besźel and Ul Qoma, occupy the same space, are culturally and politically different. Their relationship to each other is similar to that of border-sharing Cold War states. Citizens of the two cities are forbidden to interact with each other. This prohibition is radically policed. Even though the citizens of Besźel and Ul Qoma live in adjoining buildings, share roads, and walk the same streets, they are forbidden to see each other. The populations of each city grow up learning how to see what is permitted and to not see, or unsee, the forbidden other (14).I think that seeing a scatter of White Cockatoo feathers and wondering if it was by the Fox or by the Little Eagle is akin to the different practices of seeing and not seeing in Besźel and Ul Qoma. The scatter of feathers is regional and colonial and, equally, it is not. Two countries occupy the same space. Australia and a continent with its hundreds of Countries. What remains not regional is what is given and Seen as such. Understanding ourselves as walking behind everything that has gone before us enables this. As such, it is possible to see the scatter of White Cockatoo feathers as by the Fox, as happening in ‘regional Australia’, as thus characterised by around 200 years of carnage, where the success of one species comes at the expense of countless others. On the other hand, it is possible to See the feathers as by the Little Eagles, and as happening on a small patch of land in Dja Dja Wurrung Country, as a dance that has been happening for hundreds of thousands, if not millions, of years. It is a way of keeping Europe at bay.I think these Cockatoo feathers are a form of address. They are capable of interpellating something other than the regional, the colonial, and the imperial. A story of feathers, Foxes, and Little Eagles can remind us of our ‘behindness’, and evoke, and invoke, and exemplify ways of seeing and engaging with where we live that are tens of thousands of years old. This is both an act of the imagination and a practice of Seeing what is really there. 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