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A novel small-molecule arylsulfonamide causes energetic stress and suppresses breast and lung tumor growth and metastasis
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Xin Dai1, Stefan Kaluz2,4, Ying Jiang5, Lei Shi 6, DeAngelo Mckinley5, Yingzhe Wang5, Binghe Wang7, Erwin G. Van Meir2,3,4 and Chalet Tan1
1 Department of Pharmaceutics and Drug Delivery, University of Mississippi, Oxford, MS, USA
2 Department of Neurosurgery, Emory University, Atlanta, GA, USA
3 Department of Hematology and Medical Oncology, Emory University, Atlanta, GA, USA
4 Winship Cancer Institute, Emory University, Atlanta, GA, USA
5 Department of Pharmaceutical Sciences, Mercer University, Atlanta, GA, USA
6 Department of Biology, Georgia State University, Atlanta, GA, USA
7 Department of Chemistry, Georgia State University, Atlanta, GA, USA
Correspondence to:
Binghe Wang, email:
Erwin G. Van Meir, email:
Chalet Tan, email:
Keywords: arylsulfonamide; AMPK; mTORC1; HIF-1; metastasis
Received: September 04, 2017 Accepted: October 10, 2017 Published: October 29, 2017
Neoplastic cells display reprogrammed metabolism due to the heightened energetic demands and the need for biomass synthesis of a growing tumor. Targeting metabolic vulnerabilities is thus an important goal for cancer therapy. Here, we describe a novel small-molecule arylsulfonamide (N-cyclobutyl-N-((2,2-dimethyl-2H-pyrano[3,2-b]pyridin-6-yl)methyl)-3,4-dimethoxybenzenesulfonamide) that exerts potent cytotoxicity and energetic stress on tumor cells while largely sparing non-cancerous human cells. In tumor cells, it stimulates glycolysis and accelerates glucose consumption. Consequently, intracellular ATP levels plummet, triggering activation of AMP-activated protein kinase (AMPK), and diminishing the mammalian target of rapamycin complex 1 (mTORC1) and hypoxia-inducible factor 1 (HIF-1) signaling. In orthotopic triple-negative breast cancer and subcutaneous lung cancer mouse models, this arylsulfonamide robustly suppresses primary tumor growth, inhibits the formation of distant metastases to the lung, and extends mouse survival while being very well tolerated. These therapeutic effects are further potentiated by co-administration of 2-deoxy-D-glucose (2-DG), a glucose analog and glycolysis inhibitor. Collectively, our findings provide preclinical proof of concept for the further development of this arylsulfonamide in combination with 2-DG towards cancer treatment.
Uncontrolled tumor cell proliferation and growth impose tremendous demand for ATP, amino acids, lipids and nucleotides. To fuel the anabolic processes, tumor cells consume a much larger quantity of glucose than their normal counterparts and metabolize it predominantly through glycolysis even under oxygen-rich conditions, the metabolic phenotype commonly referred to as aerobic glycolysis or Warburg effect [1, 2]. In normal differentiated mammalian cells, glucose is catabolized to CO2 in a stepwise fashion via glycolysis, mitochondrial tricarboxylic acid (TCA, Krebs) cycle and oxidative phosphorylation (OXPHOS). In contrast, in tumor cells, glucose primarily undergoes glycolysis as mitochondrial oxidation is greatly reduced. Although the majority of glucose-derived carbon is converted to lactate and subsequently excreted into the extracellular space, substantial amounts of metabolic intermediaries are generated to serve as essential precursors for macromolecular biosynthesis [3].
Glycolysis occurs in the cytosol and is inefficient in terms of ATP production, as it yields only two ATP molecules per molecule of glucose, whereas the complete oxidation of each glucose molecule in mitochondria can generate up to 34 ATP molecules. A careful evaluation of the energy budget reveals that glycolysis is not the dominant producer of ATP in most tumor cell types [4, 5]. In fact, about 5% of the glucose flux still enters the mitochondria, and oxidation of metabolic intermediaries through the TCA cycle and OXPHOS contributes to about half of the total ATP production in tumor cells [1].
AMP-activated protein kinase (AMPK) is a ubiquitous energy sensor in eukaryotes that is responsive to changes in intracellular AMP/ATP ratio. AMPK acts as a metabolic checkpoint and a key regulator of energy homeostasis by promoting catabolic pathways generating ATP while switching off ATP-consuming anabolic processes, such as protein synthesis [6, 7]. Pivotal among the downstream targets of AMPK is the mammalian target of rapamycin complex 1 (mTORC1), a serine/threonine protein kinase highly conserved in all eukaryotes that coordinates cell growth with energy and nutrient availability [8]. A number of oncogenic events, such as inactivating mutations of tumor suppressor genes and hyperactivation of oncogenes converge on mTORC1 upregulation. Attenuating mTORC1 kinase activity thus represents an important objective in cancer therapy as this halts cell growth [9].
Hypoxia is a hallmark of the solid tumor microenvironment. The partial oxygen tension in normal tissues ranges from 30–60 mmHg, whereas in human tumors it is mostly below 10 mmHg (equivalent to 1.4% O2) or even less than 5 mmHg [10, 11]. Intratumoral hypoxia has long been associated with malignant phenotypes, poor prognosis and resistance to radiotherapy and chemotherapy [12]. The main mediators of biological responses to hypoxic stress in mammalian cells are hypoxia-inducible factors (HIF-1 and HIF-2), heterodimeric transcription factors composed of an oxygen-labile α–subunit (HIF-1α or HIF-2α) and a stable β-subunit (HIF-1β). By transcriptionally controlling the expression of over 100 target genes, HIFs are critically involved in tumor initiation, metabolic adaptation to hypoxia, apoptotic resistance, cancer stem cell growth, inflammatory cell recruitment, angiogenesis and metastasis [13, 14].
We have previously synthesized and characterized a series of small-molecule arylsulfonamides for their ability to block hypoxia/HIF-mediated transcription and related anticancer effects [15-20]. Structure-activity relationship optimization studies led us to develop novel arylsulfonamide N-cyclobutyl-N-((2,2-dimethyl-2H-pyrano[3,2-b]pyridin-6-yl)methyl)-3,4-dimethoxybenzenesulfonamide (abbreviated as 64B, Supplementary Figure 1) as a promising new lead compound with augmented anti-HIF potency and improved solubility [15]. Here, we report for the first time on the metabolic perturbation and anticancer efficacy of this optimized arylsulfonamide. We demonstrate that 64B alters glucose metabolism and depletes intracellular ATP, resulting in AMPK activation and diminished mTORC1/HIF-1 signaling in tumor cells. Furthermore, 64B strongly suppresses tumor growth and metastasis in triple-negative breast and lung cancer mouse models, which is further potentiated by co-administration with glycolysis inhibitor 2-deoxy-D-glucose (2-DG), supporting its further development towards cancer treatment.
64B exerts potent cytotoxicity in tumor cells and impairs their motility
We first evaluated the cytotoxicity of 64B in a variety of human and murine tumor cell lines harboring various oncogenic mutations. The IC50 of 64B was found to be in the low micromolar range in tumor cells originating from breast, lung, pancreas, skin (melanoma) and brain (glioblastoma). By contrast, 64B showed little cytotoxicity in non-cancerous breast epithelial MCF-10A and lung fibroblast MRC-5 cells (Table 1). These results provide compelling evidence that 64B is cytotoxic against tumor cells independent of tissue origin or oncogenic mutations.
Table 1: Cytotoxicity of 64B in tumor cell lines with various tissue origins and oncogenic mutations. All cells are human except where indicated.
Cell lines
IC50 (μM)
Mutated Genes
Breast cancer
Breast cancer
Lung cancer
Lung cancer
Lung cancer (murine)
Pancreatic cancer
Pancreatic cancer
Melanoma (murine)
Lung fibroblast cells
Breast epithelial cells
In colony formation assays, the number and size of colonies formed by 64B-pretreated tumor cells were clearly reduced compared to controls (Figure 1A), while those of MCF-10A and MRC-5 cells were minimally affected (Supplementary Figure 2). Cell cycle analysis showed that 64B significantly increased the G1 phase in all tested tumor cell lines (Figure 1B). Moreover, 64B increased the cell population undergoing late apoptosis (Annexin V+/PI+) by 5–10 fold (Figure 1C). These results indicate that 64B inhibits tumor cell proliferation via G1 arrest and induction of apoptosis.
Epithelial cancer cells are well known for their high motility, which is attributed to the process of epithelial-to-mesenchymal transition (EMT) [21]. In scratch wound healing cell migration assays, tumor cells in the control group showed nearly complete wound closure as illustrated by the gap area being largely filled with migrating tumor cells, whereas the treatment with 64B severely blocked wound closure (Figure 1D). In transwell matrigel invasion assays, 64B notably reduced the invading cell population compared to controls (Figure 1E). Consistent with the less motile phenotype, the protein level of epithelial cell marker E-cadherin was significantly elevated in 64B-treated tumor cells, accompanied by a reduction in vimentin and Snail1, two mesenchymal markers (Figure 1F). These results indicate that 64B impairs the motility of tumor cells.
Figure 1: 64B exerts potent cytotoxicity in tumor cells and impairs their motility. MCF-7, MDA-MB-231, A549 and LLC cells were pre-treated with 64B (5 μM) for 24 h prior to starting the assays below. A, 64B reduces colony formation of tumor cells. Colonies were visualized by crystal violet staining following 14-day incubation. B, 64B causes G1 arrest in tumor cells. Cells were stained with propidium iodide (PI) and the DNA content was analyzed by flow cytometry, the cell cycle distribution was quantified by FlowJo software. C, 64B induces apoptosis in tumor cells. Following dual staining with Annexin V-FITC and PI, cells were analyzed by flow cytometry. D, 64B impedes tumor cell migration. Scratch wounds were generated in cell monolayer and the amount of gap closure measured following 24-h incubation. E, 64B diminishes tumor cell invasion. Cells were seeded in matrigel-coated transwell chambers and incubated for 24 h, and the invading cells on the bottom of the chamber inserts were stained with crystal violet. F, Western blot analysis showing the protein levels of E-cadherin, vimentin, and Snail1 in 64B-treated tumor cells. Protein band intensities were quantified by densitometric analysis using ImageJ software. All data show representative results obtained from three independent experiments, and the results are reported as the mean ± SD (n = 3). *, p < 0.05.
64B alters glucose metabolism and depletes ATP in tumor cells
To determine whether 64B cytotoxicity is related to metabolic vulnerabilities in tumor cells, we examined whether various nutrients (glucose, pyruvate, glutamine, aspartate, serum) could rescue the cytotoxicity of 64B. Raising the glucose content in culture medium from 5.5 mM (1 g/L, physiological level, normoglycemia) to 25 mM (4.5 g/L, hyperglycemia) drastically weakened the cytotoxicity of 64B in tumor cells as evidenced by over 10-fold increase in its IC50, implying the involvement of glucose metabolism in 64B action (Figure 2A). Elevating pyruvate concentration in the culture medium also compromised the cytotoxicity of 64B as reflected by the doubled IC50. On the other hand, an increase in the level of glutamine or aspartate barely had any effect (Figure 2A), indicating that glutamine or aspartate anaplerosis cannot reverse 64B action. To further study the effect of TCA-cycle intermediaries on 64B activity, we employed the lipophilic precursors ethyl acetoacetate, dimethyl α-ketoglutarate (α-KG), dimethyl succinate and diethyl oxaloacetate (OAA) given their improved cell membrane permeability, as the alkyl groups can be hydrolyzed by intracellular esterases [22]. Although replenishing α-KG, succinate or OAA had negligible effects, the addition of acetoacetate, a precursor of acetyl-CoA, partially rescued the cytotoxicity of 64B (Figure 2A). As pyruvate is known to be oxidized to form acetyl-CoA, the commitment step into the TCA cycle [3], these results suggest a blockade in the conversion of pyruvate to acetyl-CoA in 64B-treated tumor cells.
To further investigate the link between 64B activity and altered glucose metabolism, we examined glucose consumption and energetic status in tumor cells. 64B-treated tumor cells consumed significantly more glucose than controls under both normo- and hyper-glycemic conditions, as reflected by a steeper decline in glucose levels in culture medium over a 24-h incubation period (Figure 2B). In tumor cells cultured under normoglycemia, 64B markedly reduced intracellular ATP levels within 12 h of incubation, which was further exacerbated by 24 h (Figure 2C and Supplementary Figure 3A). Such effect was absent in tumor cells grown in hyperglycemic medium. These results indicate that 64B causes intracellular ATP depletion in a glucose-dependent manner while accelerating glucose consumption.
To corroborate 64B-mediated perturbation of glucose metabolism in tumor cells, we next assessed the levels of key glucose metabolites. In tumor cells, even in the presence of abundant O2, glucose is sequentially catabolized to form pyruvate, which is predominantly reduced in the cytosol to generate lactate and NAD+, the latter serving as an important electron acceptor in both glycolysis and the TCA cycle. A small fraction of the pyruvate flux enters the mitochondria and is irreversibly oxidized to form acetyl-CoA, the commitment step into the TCA cycle [3]. We found that 64B-treated tumor cells secreted significantly higher levels of lactate into the culture medium than the controls, indicating that 64B stimulates aerobic glycolysis (Figure 2D and Supplementary Figure 3B). In tumor cells grown under normoglycemic conditions, 64B caused a significant increase in pyruvate along with a sharp decline in acetyl-CoA. The total cellular NAD(H) level was markedly decreased and accompanied by an increased NAD+/NADH ratio, reflecting a drastic decline in intracellular NADH level. The buildup of intracellular pyruvate and the concomitant reduction in acetyl-CoA and NADH is consistent with the notion that 64B treatment results in decreased entrance of pyruvate into the TCA cycle, which diminishes ATP production in the mitochondria. Of note, 64B caused negligible metabolic disturbance and energetic stress under hyperglycemia (Figure 2C, 2D), in agreement with weakened 64B cytotoxicity observed in Figure 2A.
Figure 2: 64B alters glucose metabolism and depletes ATP in tumor cells. Tumor cells were cultured in normoglycemic or hyperglycemic DMEM, and were treated with 64B (5 μM) for 24 h unless indicated otherwise. A, Cell proliferation assay showing the IC50 of 64B in MDA-MB-231 and A549 cells cultured for 72 h in the presence of 5.5 mM or 25 mM glucose, 1 mM or 10 mM pyruvate, 2 mM or 20 mM glutamine, 0.1 mM or 1 mM aspartate, 1% FBS or 10% FBS, 1 mM ethyl acetoacetate, 0.25 mM dimethyl α-KG, 1 mM dimethyl succinate and 1 mM diethyl OAA. B, Effect of 64B on glucose consumption in tumor cells by measuring the glucose level in culture medium. C, Effect of 64B on intracellular ATP level. D, Effect of 64B on the levels of metabolic intermediaries in tumor cells. All data show representative results obtained from three independent experiments, and the results are reported as the mean ± SD (n = 3). *, p < 0.05. **, p < 0.01.
64B downregulates mTORC1 signaling and disrupts HIF-1 transactivation
Depletion of intracellular ATP is known to activate AMPK, which in turn inhibits mTORC1 signaling, leading to global shutdown of protein synthesis [6]. We found strong p-AMPK induction 24 h after tumor cell treatment with 64B under normoglycemia, which was accompanied with a decrease in p-mTOR1 and its downstream effectors S6 ribosomal protein kinase 1 (S6K1), 4E-BP1 and EIF4G (Figure 3A). These effects were absent under hyperglycemic conditions.
To gauge the effect of 64B on mTORC1-dependent protein translation, we then examined the protein level of HIF-1α, the oxygen-sensitive and regulatory subunit of HIF-1, which is also a sensor of translation efficiency due to its rapid turnover [23]. In tumor cells grown in normoglycemic medium, 64B abrogated hypoxia (1% O2)-induced HIF-1α protein accumulation, whereas HIF-1β remained unaltered (Figure 3B). Consequently, this led to attenuated transactivation of HIF-1 target genes under hypoxia, as exemplified by reduced glut-1 and vegf transcription in 64B-treated tumor cells without affecting HIF-1α mRNA level (Figure 3C). Such effects were absent under hyperglycemic conditions. Together, these results demonstrate that 64B triggers the energy sensor AMPK in tumor cells and downregulates mTORC1 signaling, leading to diminished HIF-1α protein synthesis, in effect disrupting the hypoxia-induced HIF-1 transcriptome.
Glucose deprivation potentiates the anticancer activity of 64B
Given the inverse correlation between glucose abundance and 64B activity (Figures 2 and 3), we hypothesized that glucose deprivation would sensitize tumor cells to the cytotoxicity of 64B. We took two approaches to block glucose utilization in tumor cells. First, we reduced glucose availability by knockdown of glut-1 using siRNA. Glut-1 is the primary transporter that mediates glucose uptake into human epithelial cells [24]. We found that glut-1 siRNA greatly reduced Glut-1 protein expression (Figure 4A, 4D) and enhanced the cytotoxicity of 64B in tumor cells by 2-fold (Figure 4B), which resulted in a more severe reduction in intracellular ATP level (Figure 4C) and mTORC1 signaling (Figure 4D). Notably, co-treatment of glut-1 siRNA with 64B was able to sensitize hyperglycemic tumor cells to its cytotoxic effects.
Second, we used 2-DG, a glucose analog that competes with glucose uptake and inhibits glycolytic enzyme activity [25]. Again, 2-DG pronouncedly sensitized tumor cells to the cytotoxicity of 64B, and its effect was accentuated under hyperglycemia (Figure 4E). Importantly, in hyperglycemic tumor cells, the combined treatment with 64B and 2-DG significantly reduced intracellular ATP (Figure 4F) and mTORC1 signaling (Figure 4G), which was not achieved by either agent individually. Similarly, the sensitization effects of 2-DG on 64B activity were also observed in tumor cells under hypoxia (Figure 4H, 4I). Together, these results clearly indicate that glucose deprivation enhances the cytotoxicity of 64B in tumor cells.
Figure 3: 64B downregulates mTORC1 signaling and disrupts HIF-1 transactivation in tumor cells. Tumor cells were cultured in normoglycemic or hyperglycemic DMEM under normoxia (21% O2) or hypoxia (1% O2), and were treated with 64B (5 μM) for 24 h. A,B, Western blot analysis showing the effect of 64B on the levels of indicated proteins under normoxia (A) and hypoxia (B). C, qRT-PCR measuring the expression of HIF-1α, Glut-1 and VEGF mRNAs in 64B-treated tumor cells under hypoxia. All data show representative results obtained from three independent experiments, and the results are reported as the mean ± SD (n = 3). *, p < 0.05. **, p < 0.01.
Figure 4: Glucose deprivation potentiates the anticancer activity of 64B. MDA-MB-231 and A549 cells were cultured in normoglycemic or hyperglycemic DMEM, and were treated with 64B (5 μM) and/or 2-DG (2 mM) for 24 h unless indicated otherwise. A, Knockdown of Glut-1 expression by siGlut-1 was verified by immunofluorescent staining in tumor cells after 48 h. The nuclei of tumor cells were counter-stained with DAPI. B, C, Effect of the co-treatment with siGlut-1 and 64B on the cytotoxicity IC50 of 64B in tumor cells (B) and intracellular ATP level (C). Equal amount of scrambled siRNA (SCR) was used as a control. D, Western blot analysis showing effect of co-treatment with siGlut-1 and 64B on levels of indicated proteins. E, F, Effect of co-treatment with 64B and 2-DG on the cytotoxicity IC50 of 64B in tumor cells (E) and intracellular ATP level (F). G, Western blot analysis showing the effect of combined treatment with 64B and 2-DG on levels of indicated proteins. H, Effect of co-treatment with 64B and 2-DG on the cytotoxicity IC50 of 64B under hypoxia (1% O2) for 72 h. I, Western blot analysis showing effect of co-reatment with 64B and 2-DG on levels of indicated proteins under hypoxia. All data show representative results obtained from three independent experiments, and the results are reported as the mean ± SD (n = 3). *, p < 0.05. **, p < 0.01.
Combined treatment with 64B and 2-DG potently suppresses tumor growth and metastasis
We next investigated the therapeutic potential of 64B/2-DG combination in vivo. We first characterized the pharmacokinetic profile of 64B in mice bearing orthotopic MDA-MB-231 breast tumor xenografts. 64B was readily and continuously absorbed from the intraperitoneal cavity, as reflected by the sustained plasma concentration of about 3 µM observed between 15–60 min following i.p. injection (Supplementary Figure 4A). Co-administration of 2-DG minimally influenced the levels of 64B in plasma, tumor, brain and liver (Supplementary Figure 4B), indicating that 2-DG does not interfere with the pharmacokinetics of 64B.
We then evaluated the anticancer efficacy of 64B in two tumor models. In mice bearing orthotopic MDA-MB-231 triple-negative human breast tumor xenografts, daily treatment of 2-DG at an oral dose of 1 g/kg for 14 consecutive days modestly slowed down tumor growth, whereas 64B as a single agent at a daily i.p. dose of 60 mg/kg significantly retarded tumor growth (Figure 5A). The combined treatment with 64B and 2-DG further enhanced the anticancer efficacy and nearly completely arrested tumor growth. The weight of tumors harvested on day 28 confirmed the same trend (Figure 5B). The body weight of mice remained steady during the entire 4-week duration (Figure 5C), indicating no overt treatment-associated toxicities. Immunohistochemistry staining of the tumor tissues for the cell proliferation marker Ki-67 revealed that tumor cells in the 64B/2-DG-treated mice had the lowest mitotic index, while the tumors in PBS- and 2-DG-treated mice were highly proliferative (Figure 5D). Similarly, 64B/2-DG combination resulted in superior tumor growth retardation in mice bearing subcutaneous LLC tumors, a highly aggressive murine lung tumor model (Figure 5E, 5F, 5G, 5H). Treatment with 64B alone or in combination with 2-DG clearly improved the survival of LLC tumor-bearing mice (Figure 5I). Taken together, these results demonstrate potent anticancer efficacy of 64B in vivo, which can be further improved by the combined treatment with 2-DG.
Figure 5: Combined treatment with 64B and 2-DG markedly retards tumor growth. Mice received daily administration of 64B (60 mg/kg, i.p.) and/or 2-DG (1 g/kg, p.o.) for indicated duration (n = 5 mice/group). A, Treatment scheme (upper) and tumor growth curves (lower) in mice bearing orthotopic MDA-MB-231 breast tumors. The mice were treated for 14 days and sacrificed on day 28. B, Image and weight of MDA-MB-231 tumors harvested at termination. C, Average body weight of mice bearing MDA-MB-231 tumors during the entire course of the study. D, H&E staining and Ki-67 immunostaining of the MDA-MB-231 tumor sections. Images were acquired at 40× magnification. Quantification of Ki-67-positive cells was determined microscopically (mean cell number per field ± SD, 12 fields per tumor). E, Treatment scheme (upper) and tumor growth curves (lower) in mice bearing subcutaneous LLC tumors. The mice were treated for 10 days and sacrificed on day 12. F, Image and weight of LLC tumors harvested at termination. G, Average body weight of mice bearing LLC tumors during the entire course of the study. H, H&E staining and Ki-67 immunostaining of LLC tumor sections. Images were acquired at 40× magnification. Quantification of Ki-67-positive cells was determined microscopically (mean cell number per field ± SD, 12 fields per tumor). I, Kaplan-Meier survival curves in mice bearing subcutaneous LLC tumors. Mice were euthanized when the tumor size reached IACUC termination criteria.The results are reported as the mean ± SD (n = 5). *, p < 0.05. **, p < 0.01.
Finally, as metastasis is the major cause of death in patients, we determined the impact of 64B on tumor metastasis in these two tumor models, both of which are highly prone to lung metastasis [26-28]. Orthotopic MDA-MB-231 triple-negative breast cancer-bearing mice developed an average of 8 metastatic tumor nodules in the lung on day 28 post-tumor cell inoculation (Figure 6A). Treatment with 64B alone or in combination with 2-DG completely prevented tumor nodule formation in the lung, while 2-DG individually reduced the lung metastases by half. The mesenteric lymph nodes in PBS- and 2-DG-treated mice were noticeably enlarged due to their colonization by of proliferating tumor cells (Figure 6B). In contrast, those from 64B- and 64B/2-DG-treated mice maintained a normal morphology and showed over 75% reduction in Ki-67 positive cells (Figure 6C). Similarly, complete blockade of lung metastasis was observed in mice bearing subcutaneous LLC tumors that received 64B treatment individually or in combination with 2-DG (Figure 6D), and the mesenteric lymph nodes in these mice were much less colonized with proliferating tumor cells (Figure 6E, 6F). These results provide strong evidence that 64B suppresses tumor metastasis in vivo.
Figure 6: Combined treatment with 64B and 2-DG suppresses tumor metastasis. A, H&E staining and Ki-67 immunostaining of the lung tissue sections in mice bearing orthotopic MDA-MB-231 tumors (sacrificed on day 28). Metastatic tumor nodules were counted in the five lobes of the lungs and the quantification is shown on the right. B, Images of the mesenteric lymph nodes harvested from mice bearing orthotopic MDA-MB-231 breast tumors. C, H & E staining and Ki-67 immunostaining of the mesenteric lymph node tissue sections in mice bearing orthotopic MDA-MB-231 tumors. Quantification of Ki-67-positive cells was determined microscopically (mean cell number per field ± SD, 12 fields per lymph node) and is shown on the right. D, H & E staining and Ki-67 immunostaining of the lung tissue sections in mice bearing subcutaneous LLC tumors (sacrificed on day 12). Metastatic tumor nodules were counted in the five lobes of the lungs and the quantification is shown on the right. E, Images of mesenteric lymph nodes harvested in mice bearing subcutaneous LLC tumors. F, H & E staining and Ki-67 immunostaining of the mesenteric lymph node tissue sections in mice bearing subcutaneous LLC tumors. Quantification of Ki-67-positive cells was determined microscopically (mean cell number per field ± SD, 12 fields per lymph node) and is shown on the right. The data are reported as the mean ± SD (n = 5). *, p < 0.05. **, p < 0.01.
Tumor cells undergo pronounced metabolic reprogramming to engender macromolecular building blocks that are essential for rapid tumor cell proliferation and growth. As a master regulator for the biosynthesis of proteins, lipids and nucleic acids, mTORC1 plays a central role in coordinating anabolic pathways that ultimately drive tumor growth. Two of the best characterized mTORC1 immediate targets, S6Ks and 4E-BPs, are not only critically involved in protein synthesis by increasing ribosomal biogenesis and mRNA translational initiation, but also regulate lipogenesis and nucleotide synthesis [8, 9]. mTORC1 regulates translation of the mRNA encoding HIF-1α, the α subunit of hypoxia-inducible transcription factor HIF-1. Hypoxia is emerging as a key microenvironmental factor in the regulation of tumor metastasis, largely through HIF activation. Numerous HIF-1 target genes mediate invasiveness and the metastatic cascade during tumor progression [14]. Elevated HIF-1α protein levels in solid tumor biopsies correlate with metastasis in patients [29, 30], and inactivation of HIF-1 reduces the metastatic potential of tumor cells in animal models [31, 32].
We have had a long-standing interest in HIF-1 signaling and reported previously the synthesis and characterization of a series of arylsulfonamide compounds that potently and selectively inhibit the HIF-1 pathway [15-20]. Among them, N-((2,2-dimethyl-2H-chromen-6-yl)methyl)-3,4-dimethoxy-N-phenylbenzenesulfonamide (abbreviated as KCN1) markedly inhibits tumor growth and HIF activity in mice with little toxicity towards normal cells and tissues [17]. Recently, we have synthesized and screened more analogs of KCN1, among which 64B is found to be more potent than KCN1 (IC50~0.3 vs. 0.6 μM) in a HIF-dependent cell-based luciferase reporter assay [15]. In the present study, we investigated the anticancer activity and mechanism of 64B in vitro and in vivo. We show that 64B downregulates mTORC1/HIF-1 signaling in tumor cells under glucose-limiting conditions. Under normoglycemic cell culture conditions, 64B interferes with glucose metabolism in tumor cells and elicits energetic stress that triggers AMPK activation. In tumor cells, the vast majority of glucose influx is known to be biotransformed into lactate and excreted into the extracellular space; yet, some glucose flux to the TCA cycle remain indispensible because of its crucial role in generating metabolic intermediaries and intracellular ATP [33]. We demonstrate that 64B elevates lactate production in tumor cells while reducing the conversion of pyruvate to acetyl-CoA, which hinders TCA flux and ensuing ATP production. Further studies are needed to elucidate the molecular mechanism by which 64B disrupts of pyruvate flux into the TCA cycle.
The metabolic and energetic stress caused by 64B is further potentiated by co-treatment with 2-DG, a glucose analog and glycolysis inhibitor that has been in clinical trials for cancer treatment [34-36]. Synergistic anticancer activities have been reported when 2-DG is combined with small-molecule inhibitors targeting the TCA cycle or OXPHOS [37, 38]. By dual inhibition of glycolysis and mitochondrial respiration, more severe energetic stress can be achieved in tumor cells. Our in vitro studies clearly evidence that 64B is highly effective at inhibiting ATP production in cultured tumor cells under glucose-limited growth conditions or pharmacologically-induced glycolysis inhibition. Results of in vivo investigation further demonstrate that 64B possesses remarkable anticancer efficacy in animal models for cancer. 64B notably inhibits tumor growth when administered as a single agent or in combination with 2-DG in orthotopic MDA-MB-231 triple-negative breast cancer and subcutaneous LLC lung cancer mouse models, both of which are known for highly aggressive tumor growth and metastatic propensity to the lung. While having little efficacy as a monotherapy, 2-DG markedly augments tumor growth retardation by 64B, probably owing to a synergy in blocking tumor cell ATP production.
Most importantly, 64B exhibits strong anti-metastatic effect in vivo, completely blocking metastatic tumor nodule formation in the lung. The significant reduction in tumor burden at the primary site in 64B-treated mice likely minimizes tumor cell extravasation into the circulation. Whether 64B also impedes colonization in the distant organ remains to be established. EMT is an enabling step towards tumor metastasis, and is characterized by upregulation of mesenchymal markers such as intermediate filament protein vimentin and reduced expression of the normal epithelium adhesion protein E-cadherin [21]. Snail1 is a transcription factor that binds the promoter region of CDH1, encoding E-cadherin, to repress its transcription. Consistently, 64B-treated tumor cells display impaired migratory and invasive potential, which is accompanied by decreased protein levels of Snail1 and vimentin as well as an increase in E-cadherin expression. The translation of both vimentin and Snail1 mRNAs, along with a number of other pro-invasion messenger RNAs, is controlled by mTORC1 [39, 40], likely explaining their inhibition and restoration of E-cadherin in 64B-treated tumor cells.
We demonstrate that the anticancer activity of 64B is inversely affected by the glucose content to which tumor cells are exposed, and glucose deprivation potentiates 64B activity. It has been previously reported that glucose level influences the cytotoxicity of metformin, a Complex I inhibitor that halts ATP production by disrupting electron transport during mitochondrial respiration [41, 42]. Our data lend further support to prior findings that elevated glucose levels negate therapeutic interventions that target tumor glucose metabolism. Although growth media such as DMEM with high glucose content (4.5 g/L, 25 mM) are routinely used to cultivate tumor cells in vitro, they induce a metabolic phenotype that is drastically different from the much less permissive growth environment found within solid tumors [43]. Our findings underscore the importance of employing physiologically relevant glucose levels in cell cultures when evaluating anticancer agents that exploit glucose metabolic vulnerabilities.
In summary, our findings demonstrate that 64B potently suppresses tumor growth and metastasis in mouse models of triple-negative breast cancer and lung cancer. 64B perturbs tumor metabolism resulting in depletion of intracellular ATP and attenuation in mTORC1-mediated translation of critical pro-tumorigenic proteins including HIF-1 and EMT mediators that are critical for tumor growth and metastasis. The anticancer efficacy of 64B is further potentiated by glucose deprivation with 2-DG, compounding the metabolic and energetic stress exerted on tumor cells. Altogether, these data support further development of arylsulfonamides as a novel class of anticancer agents targeting metabolic vulnerabilities in cancer, especially in combination with a glycolysis inhibitor.
MATERIALS and METHODS
Cell lines and reagents
MCF-7, MDA-MB-231, A549, NCI-H1975, Lewis lung cancer (LLC), BxPC-3, Panc-1, SK-MEL-28, B16-F10, LN229, MRC-5 and MCF-10A were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Unless otherwise indicated, MCF-7, MDA-MB-231, A549, Panc-1, and MRC-5 were grown in DMEM (Life Technologies, Grand Island, NY) with 5.5 mM glucose (normoglycemic medium). LLC, SK-MEL-28 and LN229 were grown in DMEM with 25 mM glucose (hyperglycemic medium). NIH-H1975 and BxPC-3 were grown in RPMI 1640 medium, which contains 10 mM glucose (Life Technologies). All these media were supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Life Technologies). MCF-10A were cultured in DMEM/F12 (25 mM glucose) supplemented with 5% FBS, 20 ng/mL epidermal growth factor (EGF) (R&D Systems, Minneapolis, MN), 10 μg/mL insulin, 0.5 mg/mL hydrocortisone, 100 ng/mL cholera toxin (Sigma, St. Louis, MO) and 1% penicillin/streptomycin. All cells were cultured at 37°C in a 5% CO2 incubator. For the hypoxia conditions, cells were kept under 1% O2 in a modular incubator chamber (Billups-Rethenberg, Del Mar, CA) at 37°C.
64B was synthesized and characterized as reported in our previous work [15]. For all in vitro studies described below, a stock solution of 64B in DMSO was used to dilute to the final concentrations in the culture medium with DMSO concentration no more than 0.5% (v/v).
Cell proliferation assay
Cells were seeded at a density of 5,000 cells/well in 96-well plates and treated with 64B (1–100 μM), and their viability was determined after 72-h treatment. To study the effect of Glut-1 knockdown on the cytotoxicity of 64B, cells were transfected with Glut-1 siRNA for 24 h by Lipofectamine 2000 (Life Technologies) according to the manufacturer's protocol, followed by treatment with 64B (1–100 μM) for 48 h. To study the combination effect of 64B and 2-DG, cells were treated with 64B (1–100 μM) in the presence of 2-DG (2 mM) for 72 h. Subsequently, cells were fixed with 1% glutaraldehyde, stained with 0.1% crystal violet, and the crystals dissolved in 10% acetic acid. The absorbance was quantified at 595 nm on a FLUOstar Omega plate reader (Cary, NC). The relative cell viability was calculated as the percentage of absorbance of the drug-treated vs. the untreated controls.
Colony formation assay
Following pre-treatment with 64B (5 μM) for 24 h, cells were seeded in 12-well plates at a density of 150 cells/well, and were incubated without 64B for another 14 days. The adherent cell colonies (with a minimum of 50 cells per colony) were fixed with 1% glutaraldehyde, stained with 0.1% crystal violet, imaged, and counted using an inverted microscope (Olympus, Tokyo, Japan).
Cell cycle analysis
In 6-well plates (1.0 × 106 cells/well) cells were treated with 64B (5 μM) for 24 h, and then fixed in 70% ethanol at 4°C overnight, washed with cold PBS, re-suspended in a staining solution 0.1% Triton-X-100, 50 μg/ml propidium iodide (PI) and 1 mg/ml RNase A for 30 min at room temperature, and analyzed by a BD Accuri C6 Flow Cytometer System (San Jose, CA). The DNA content distribution was analyzed using the FlowJo 9.3.1 software (Tree Star, Ashland, OR).
Apoptosis assay
In 6-well plates, tumor cells (1.0 × 106 cells/well) were treated with 64B (5 μM) for 24 h and stained with Annexin V-FITC and PI (BD Biosciences), which were analyzed by flow cytometry.
Measurement of metabolic intermediaries of glucose
Tumor cells (1.0 × 106 cells/well) were seeded in 6-well plates containing DMEM with 5.5 mM or 25 mM glucose. After 24 h, the medium was replaced with 2 mL fresh medium containing 64B (5 μM) and the cells were incubated for up to 24 h. Cells were lysed, and intracellular metabolites were measured using commercial kits detecting ATP (Abcam, Cambridge, MA), pyruvate (Sigma-Aldrich), acetyl-CoA (PicoProbe, Abcam) and NAD+/NADH ratio (BioVision, Milpitas, CA), respectively, according to the manufacturers' recommended procedures. To measure pyruvate production, cells were cultured in pyruvate-free medium. Levels of glucose and lactate in the conditioned medium were analyzed by using a glucose colorimetric assay kit (Sigma-Aldrich) and an EnzyChromTM L-Lactate assay kit (BioAssay Systems, Hayward, CA), respectively.
Western blot analysis
Tumor cells were lysed using RIPA buffer supplemented with protease and phosphatase inhibitors. Cell lysates containing 40 μg of total proteins were resolved in 12% SDS−PAGE gels, and transferred onto nitrocellulose membranes (GE Healthcare, Pittsburgh, PA). Blots were probed with antibodies targeting the following proteins: p-mTOR, mTOR, p-AMPKα (Thr172), AMPKα, p-S6K1 (Thr398), S6K1, p-4E-BP1 (Thr37/46), 4E-BP1, p-EIF4G (Thr1108) and EIF4G (Cell Signaling, Danvers, MA); Snail1 and β-actin (Santa Cruz, Dallas, TX); HIF-1α and HIF-1β (BD Biosciences) and Glut-1 (Abcam). All antibodies were used at 1:1000 dilution. Protein band intensities were quantified by densitometric analysis using ImageJ software (NIH, Bethesda, MD).
Quantitative real-time PCR (qRT-PCR) analysis
Tumor cells were lysed in Trizol (Invitrogen) for total RNA extraction. The expression of mRNAs of HIF-1α, Glut-1 and VEGF was analyzed by qRT-PCR as previously described [44]. The sequences of the primers are listed in Supplementary Table 1.
Animal experiments
All animal procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The Institutional Animal Care and Use Committee at Mercer University approved the protocol (Approval Number: A1105007). For the orthotopic breast tumor model, MDA-MB-231 human cells (1 × 106 in 0.1 mL PBS) were injected into the fourth mammary pad of 6-week old female athymic nude mice (nu/nu, Charles River, Wilmington, MA). When the tumor size reached 50–100 mm3, the mice were randomized into four treatment groups (n=5 mice/group): (A) PBS control; (B) 2-DG; (C) 64B; (D) 64B/2-DG combination. 2-DG was orally (p.o.) administered to the mice at 1 g/kg once daily, while 64B was dissolved in Cremophor RH40:PEG400:deionized water at 3:1:8 (v/v/v) and intraperitoneally (i.p.) injected into the mice at 60 mg/kg once daily. For the mice receiving the 64B/2-DG combination therapy, the oral dosing of 2-DG was given 30 min prior to the i.p. administration of 64B. The mice were treated for 14 consecutive days. The tumor size was measured with a caliper twice weekly along with the mouse body weight, and the tumor volume was calculated as 1/2 × length × width2. For the subcutaneous lung tumor model, LLC cells (5 × 105 in 0.1 mL PBS) were injected into the axillary region of syngeneic 6 week-old male C57BL/6 mice (Charles River). When the tumors were palpable, the mice were randomized into four treatment groups (n=5) as described above.
To observe tumor metastasis to the lung and lymphatic tissues, the tumor xenografts, lungs and lymph nodes in the tumor-bearing mice were excised for immediate formalin fixation. Formalin-fixed and paraffin-embedded tissue specimens of 3- to 4-μm thickness were sectioned and stained with hematoxylin and eosin (H&E) and a Ki-67 antibody (Cell Signaling) following standard immunohistochemistry protocols.
Statistical analysis
All data were presented as the mean ± SD. Data from different groups were compared using Student's t-test. A p-value of less than 0.05 was considered statistically significant.
2-DG, 2-deoxy-D-glucose; 64B, N-cyclobutyl-N-((2,2-dimethyl-2H-pyrano[3,2-b]pyridin-6-yl)methyl)-3,4-dimethoxybenzenesulfonamide; α-KG, α-ketoglutarate; AMPK, AMP-activated protein kinase; EMT, epithelial-to-mesenchymal transition; HIF-1, hypoxia-inducible factor 1; KCN1, N-((2,2-dimethyl-2H-chromen-6-yl)methyl)-3,4-dimethoxy-N-phenylbenzenesulfonamide; mTORC1, mammalian target of rapamycin complex 1; OAA, oxaloacetate; OXPHOS, oxidative phosphorylation; PI, propidium iodide; TCA, tricarboxylic acid.
Authors' contributions
XD, YJ, LS, DM and YW conducted all experiments. XD and CT analyzed the data and wrote the manuscript. XD, SK, BW, EGVM and CT conceived the project and contributed to manuscript writing.
CONFLICTs OF INTEREST
EGVM and BW have ownership interest as co-inventors on a patent jointly owned by Emory University and Georgia State University. The other authors declare that they have no competing interests.
This work was supported in part by grants from the NIH (R01 CA116804, R01 CA176001, R01 CA180805, and P30 CA138292), a Winship Cancer Institute pilot grant, the V Foundation, the Max Cure Foundation, the Samuel Waxman Cancer Research Foundation, and the Alan B. Slifka Foundation.
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text:
MAY in instagrams ♥
Hi everyone, can you believe it? It's already the last day of May! This month went by so fast!! I want to share all my instagram-posts of the month May today, eventhough it wasn't a really good instagram-month for me. Anyways, I hope you like it!!

subset: commoncrawl


text:
Depending on the printer you are using, an administrator password is specified at the time of purchase.
The password is either "canon" or the printer serial number if it is specified.
Model whose administrator password is specified as "canon":
MG7500 series, MG6700 series, MG6600 series, MG5600 series
MG2900 series, MX490 series, MB5300 series, MB5000 series
MB2300 series, MB2000 series, E480 series, E460 series
iP110 series, iB4000 series, PRO-100S series, PRO-10S series
Set the password by using 0 to 32 characters.
Model whose serial number is used as the administrator password:
Where Serial Number Is Located
Set the password by using 4 to 32 characters.
operation panel of the printer
For more on initializing the printer, search for "UG067" or "UG505" on your printer's Online Manual and see the instructions shown.


text:
Here is a spring market basket from the produce section of our local store. When I am looking for ideas for what to cook I usually start shopping in the produce section because it inspires me. Left to right: baby romaine, red pepper, ugly tomato, golden beet, Easter Egg radishes, asparagus and an artichoke.
I decided to make a vegetarian dinner :
- Spring root vegetable salad
- Boiled artichoke with lemon
- Roasted vegetables over whole grain couscous
First, I started the artichoke since it takes the longest to cook:
Trim the spikes with scissors and cut the artichoke in half. Remove the center:
Place in boiling water and cook until tender, about 10-15 minutes. They tend to float so you should flip them half way:
Here are the finished artichoke halves - they are served on a white platter with fresh lemon.
The eggplant, asparagus and red peppers were cut in thin sticks. I lightly brushed them with olive oil and finely minced garlic:
And then broiled them in the oven for 10 minutes - look how wonderfully golden they came out:
Whole grain couscous is perhaps the easiest whole grain to prepare. It involves 2 ingredients and a minute of time:
1 cup water - bring to boil in a large measuring cup in the microwave
1 cup whole grain couscous - pour it into the boiling water, sit one minute, fluff with a fork. You can toss with lemon and herbs or a light dressing. It can go into salads or be a side dish. In this case it was the main entree with roasted vegetables on top:
When the vegetables were golden brown, they were removed from the oven and placed atop the fluffy whole grain couscous:
Meanwhile, while the veggies were cooking I prepared the spring root salad. Here are the Easter Egg radishes and the golden beet:
I shredded the beet with the grater and finely sliced the radishes atop the baby romaine. They were presented on a flat platter :


text:
Building for the Future: Easing California's Transportation Crisis with Tolls and Public-Private Partnerships
Publication Date: March 2005
Publisher(s): Reason Foundation
Author(s): Robert W. Jr. Poole
Topic: Transportation (Highways)
Coverage: California
California is projected to grow to 50 million people between 2000 and 2030, a net increase of 16 million. California's urban freeway systems are already nearing capacity, with massive congestion during ever-lengthening peak periods, yet current plans call for only minor increases in capacity. Since the freeways will continue to carry more than 90 percent of commuters and the vast majority of all freight, it is crucial that California's highway capacity keep pace with growth, so that people and goods can keep moving. But even before the current transportation funding crisis, available highway financial resources were hard-pressed even to maintain the existing infrastructure, let alone add to its capacity. Hence, this study proposes the creation of new financing and project-delivery methods for California highways, aimed at providing the resources to make major improvements in this vitally important infrastructure.


text:
Literature & Thought
Teach students to be critical thinkers through a unique questioning strategy supporting close reading of complex texts
The unique design of Literature & Thought provides the thematic literature and teaching support you need.
- informational text
- close reading strategies
- writing to sources
- critical thinking
- appropriate text complexity
- text dependent questions and tasks
Choose from 25 titles in historical events and eras, literary themes, literary genres, government and current events.
Student Editions
- A variety of thematically-connected test types—outstanding literature, literary nonfiction, and content-rich informational texts engage students and focus attention on critical thinking questions.
- Creating context—an introductory essay, graphics, and concept vocabulary support the essential question and help students connect with the overall theme.
- Essential questions (whole book) and cluster questions (units) focus on developing specific critical thinking skills through careful reading, textual analysis, discussion, and writing activities.
- Responding to cluster activities—textual analysis, writing, and discussion activities follow each cluster. The final cluster focuses on synthesizing information to answer the essential question.
Close Reading Skills Handbook—detailed guidance to help students turn standards into actionable skills practice.
- Close Reading guidelines for a careful interpretation of the text
- specific to the type of text being read—informational, argumentative, narrative, or poetry
- students are prompted with detailed "questions to ask" and "where to look for answers" for a guided first read and focused re-reading
- Textual Evidence—guidelines show students how to work textual evidence into a written literary analysis or informational report.
- Comparing Texts—focus points and questions for comparing and contrasting different texts with an emphasis on strategies if text types or medium differ.


text:
Circles Project
In May of 2008, on a trip to Arizona, I found myself photographing beautiful circular patterns in sandstone. I mentioned to a friend that I loved the simplicity of natural circles, and their sometimes unexpected appearance in nature. She suggested that I start a "Circles Project".
In the back of my mind, I also thought about a book I read to my children a thousand times when they were small – the quiet, beautiful story of a little girl who lacked confidence in her own artistic ability – The Dot – by Peter H. Reynolds. If you haven't read it – I highly recommend it. The lesson rings true no matter your age, and the book is one of just a few that I'll keep in my own personal collection even after my children are grown.
(For those who are interested, Mr. Reynolds has written other excellent books as well. Another favorite of mine is titled "Ish." Again, these are children's stories… but no less relevant because they were meant for a young audience. I've met thousands of adults who struggle with the same issues.)
Anyway – with that story and my friend's suggestion in mind, I set out to capture a series of natural circles. I've been shooting circles for more than three years… ever since that trip to Arizona… and I'll continue to add to the collection over time. Even now, after all this time, there is something about natural circles that reminds me to simplify… my life, my composition, my thought processes.
Personal projects are a great way to keep the creative juices flowing.. and to focus your photographic vision over time. Although I am always shooting other things as well – shooting circles is like adding to those fun collections we had as kids. Bottle caps or stamps or coins. But there's something of myself in this collection, too. It's more personal.
You can see that my circle images are pretty different from one another. Tiny pebbles in circles carved from solid rock… mud bubbles formed as gas escapes from underground… patterns in the sand that were created as grasses moved in the wind… lichen growing on rocks…
The variety is part of what makes the project so appealing to me. You start with a single image – a single idea – and pretty soon, you start seeing things in an entirely different light. I find circles in places I wouldn't expect to find them!
I love finding another circle to add to my collection. So why not create your own project or assignment. Think about what appeals to you – something that interests you or defines you in some way. Take a simple theme and spin it – I think you'll find that it becomes far more than you expected!


text:
Written evidence suggests that people have used globes to model the world around them since antiquity; Strabo (63/64BCE-24CE) reported that Crates of Mallos had a globe of the equivalent of 10 feet in diameter. Globes are delicate, though, and the surviving evidence for early globe use is sparse. The earliest globe that survives today was made in 1492 by Martin Behaim, a German navigator and geographer in the employ of King João II of Portugal. Behaim's globe recorded not only the lie of the lands being discovered by seabourne explorers, but also details of overseas commodities, market places and local trading protocols. Thus, the earliest surviving globe, which probably reflects many others produced around the same time, features information on more than cartography.
Globes retained appeal to a wide range of audiences into the 16th century. Gemma Frisius, a Dutch cartographer, mathematician and instrument maker who worked at Louvain, wrote in his Principiis de astronomiae et cosmographiae (1530) that:
[T]he mounted globe ... is the only one of all instruments whose frequent usage delights astronomers, leads geographers, confirms historians, enriches and improves legists [les legists], is admired by grammarians, guides pilots, in short, aside from its beauty, its form is indescribably useful and necessary for everyone.
Frisius' account of globes as beautiful as well as useful is significant - globes have tended to attract attention as art objects, as well as for their depiction of geographically, politically and economically important lands. Globes were often exchanged as gifts among important rulers, since they signified command of the world, but were also suitably stately for presentation to powerful figures.
The variety of uses to which globes could be turned is illustrated by the case of a globe made by Gerard Mercator, a Flemish cartographer who trained under Frisius. Mercator included rhumb lines on his globe of 1541, which meant that the globe could have been of some use in navigational instruction. However, Mercator made the globe for Nicolas Perrenot de Granvelle, an important figure in the privy council of Emperor Charles V. De Granvelle would have been attracted by the possibility of possessing the world symbolically by possessing the globe, and the courtly setting suggests that the beauty and grandeur of the globe would have been as important as its utility.
There are no 16th-century globes in the Whipple collection, but the objects presented here serve to demonstrate the variety of meanings that globes have attracted.
Globes retained their decorative function in the 17th century, and some innovative designs were produced that promoted the gentlemanly use of globes as accessories or furniture items. Pocket globes were first produced in England by Joseph Moxon XR (1627-1691) in 1673, and gentlemen might well have used these miniature instruments as status symbols. Moxon also collaborated with Roger Palmer XR to make the 'English Globe' in 1679 (Image 1), which was best used in the garden. Indeed, the fact that the 'English globe' could not be rotated on its stand meant that it could only be used for latitudes matching that of the south of England and would have been useless on sea voyages. Pocket globes and instruments such as the 'English globe' meant that makers could promote globes among new audiences.
» Read more about pocket globes
» Read more about the "English globe"
While makers innovated some designs to attract new users, they kept abreast of cartographic developments and continued to produce globes that recorded the most recent discoveries. Robert Morden, who collaborated with William Berry and Philip Lea to produce a globe sometime between 1683 and 1690, declared in his Geography rectified, or, A description of the world (1693) that:
[I]t is a boldness justifiable by truth, to affirm that all former Geographies diligently compared with the more accurate Observations and Discoveries of late years are greatly defective, and strangely erroneous.
While Morden was not advocating the use of the globe in practical navigation, he certainly felt that accuracy was important. If a globe recorded the latest discoveries in the way that Morden described, possession of the globe would indicate the owner's awareness of the most cutting edge geographical findings.
» Read more about the globe made by Robert Morden
The 18th century is sometimes seen as a second 'age of exploration', following the initial voyages of discovery in the late 15th and 16th centuries. When four Royal Navy ships were wrecked off the Scilly Isles in 1707 on account of navigational errors, the British Government established the Commissioners for the Discovery of the Longitude at Sea (also known as the Board of Longitude), which offered large monetary prizes to anyone who could devise a method for accurately finding longitude G . This motivated a more practical approach to sailing in the 18th century.
Globes sat awkwardly among air pumps and microscopes, the established apparatus of the rationalists. Many judged that globes could only be used to record, but not to make, discoveries. For example, Joseph Priestley (1733-1804), a Fellow of the Royal Society, felt that globes were only explanatory devices, which "like books, have no uses more extensive than the view of human ingenuity".
In spite of the cynicism that some felt towards globes, they proved useful for recording the new discoveries made in the 18th century, including many of the Pacific islands and New Caledonia. Existing territories were also updated in line with continuing exploratory ventures. For instance, California, which had been depicted as an island since the early 17th century, was mapped as a peninsula with increasing frequency throughout the 18th century.
Globes had traditionally had some role in the training of navigators and astronomers and the use of the instruments in education underwent a major expansion in the 19th century. Strategies to bring the globe into the world of the child included making smaller versions and using the objects as part of a game.
Pocket globes could have provided small, if not accurate, representations of larger globes at relatively low cost, so that children could learn about the various rings on globes, or about the approximate arrangement of the continents. Many globes designed for children's education were comstructed from various pieces in the style of 3-D jigsaw puzzles. Other globe makers concentrated on trying to make their globes as portable as possible, so that the objects could be carried from classroom to classroom or between the homes of different pupils in the case of travelling tutors. John Betts' XR umbrella globe (Image 2) provided one way to do this.
» Read more about pocket globes
» Read about a jigsaw puzzle globe
Beyond the specific area of globes, the 19th century saw the development of many technologies that we take for granted today. Among these innovations were electric lighting for streets and for the home, and the telegraph cable. Now that light could be reliably provided throughout the day and night, and locations across the seas reached without arduous travel, many of the lessons that globes had been designed to teach became redundant.
For instance, it became less important that students could work out how many hours of daylight there would be at a given latitude on a particular day of the year. Globe design reflected this change in function, as the horizon ring G fell from common use towards the end of the century. Prior to this, from around the middle of the century, many makers abandoned the use of constellation figures on celestial globes in an attempt to appeal to the self-consciously scientific spirit of the age.
Technological advances in the 20th century acted on globes in a variety of ways. On one hand, the development of radio based navigation techniques in the 1940s and 1950s meant that globes were no longer used for navigation. As electric lighting had made the demonstration of solar phenomena that terrestrial globes offered less compelling, so radio navigation brought an end to the widespread use of celestial globes to show man's position relative to the sphere of fixed stars.
» Read about an early 20th-century globe used for navigating at sea
New technologies also affected perceptions of globes positively, though. When satellite pictures showing the earth's coastlines became available in the 1960s, the view that had been constructed by compiling evidence accumulated over centuries of exploration was visually confirmed for the first time. The move towards large-scale production of globes in the 20th century, for instance by the Replogle company, also meant that the public were more exposed than ever to the instruments that had been the preserve of princes and merchants 500 years before.
Katie Taylor
Katie Taylor, 'A brief history of globes', Explore Whipple Collections, Whipple Museum of the History of Science, University of Cambridge, 2009 [http://www.hps.cam.ac.uk/whipple/explore/globes/abriefhistoryofglobes/]


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Michigan State University Archives & Historical Collections (UAHC) is responsible for preserving the rich heritage and legacy of Michigan State University. This could not be accomplished without the generous support of alumni, friends, and donors.
Your history is our country's history. Letters from relatives, a grandparent's diary, old family photographs, and other materials collected over the years provide unique information about your life and the history of your family. These documents may be important to the local community. When you donate personal or family papers to an archive, your family history becomes a permanent part of the community's—and the nation's—collective memory. Your support improves the quality of our collections.
If you wish to donate personal papers, photographs, scrapbooks, audio-visual materials, or other in-kind materials, please contact Sarah Roberts, acquisitions archivist of the UAHC.
Sarah Roberts
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MSU Film and Video Preservation Fund
Save Spartan memories forever with your gift!
We have hundreds of film reels and videotapes documenting athletics and campus events from the 1930s – 1990s.
This collection must be restored and digitized soon – before the content is lost to time.
You can help preserve this Spartan history with a gift to the MSU Film and Video Preservation Fund!


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To keep up to date please download the Show My Homework App which is free. You will then be sent push notifications to let you know what homework has been issued to your child. All S1 - S4 pupils have been given a letter for parents/guardians with parental login details attached.
Solar Bear Theatre Company
St Roch'students have been working with 'Solar Bear Theatre Company' over the last year to help them develop their new show 'The Arrival' based on Shaun Tan's graphic novel of the same name. This week 'Solar Bear' returned to give us a world premier of their show before it ngoees on tour all over Scotland. This included a full day of theatrical events - characters turning up in classrooms with suitcases looking for directions, missing posters appeared in hallways and mysterious animals were vanishing. This was a fantastic partnershoip between Solar Bear and St Roch's and we hope to continue this work with the theatre company's different shows overthe next few years.
S1 and S2 Internet Safety
Faculty of Business Education and Computer Science pupils working with our new campus policeman.
St Roch's new Football Pitch
Our amazing new 3G football pitch is nearing completion. It measures in at 100m by 60m and meets SFA regulations/standards which means it meets the requirements to play professional football games on it. It is estimated to be finished by the Easter holidays. We are all very excited to use this new pitch and today we got to try it out for the first time.
The workers still have some work to do such as laying a couple more rolls of grass, cut in the marking for the pitch and they also need to spread 30 tons of sand across it and put in rubber pellets to make the grass not slippy and safe for us to use.
All of the workers have worked extremely really hard to make this wonderful plan work and they are doing a great job. The workers let us sit on some of the machines to see how some of the work might look from their point of view.
In the end we had so much fun learning from the project manager and the workers. We got to try it out for the first time today and we had loads of fun. We are looking forward to seeing the football pitch coming to life.
Saint Roch's Parent Council fun day pics
Senior Deaf students on vocational training at RSBi
Fair Trade Ambassador - Mrs McEachen
Parent Council - Mrs Hewitt receiving the Lord Provost award
Broadcast Operator Apprenticeships @ BBC (Various Locations)
UK media jobs & paid internships in TV (runners, production crew) Journalism (editorial assistants, reporters), PR & more. Advice for graduates & students.
If your applying for for a full time course commencing 28 August 2017 applications will open from January 25th 2017. see link below
TESCO Bags of Help - Grant Ref No: 7069
Congratulations! Congratulations! Congratulations!
WOW, we came first and will get £12,000
"We are delighted to let you know that following the Tesco Bags of Help vote in your local Tesco stores, your project came first and has been chosen by Tesco Customers to receive a grant award of £12,000.00!"
Voting was held at Tesco stores between Monday 31st October 2016 and Sunday 13th November 2016 and proved to be hugely successful with over 6 million votes being cast by Tesco Customers across England, Scotland and Wales.
Meet the School Parent Council
Funding we have secured and paid out over the last year or so
Funding Secured:-
£4000 secured to assist with London trip June 2015
Successful Table Top sale. Raised just over £300 for Costa Rica Trip – November 2015
£100 secured to assist with the Senior Citizen Christmas Lunch December 2015
£400 secured for Panto Trip December 2015
£500 secured to assist with Grease Production – June 2016
£4000 secured to assist with the Costa Rica Trip – June 2016
£2500 secured to assist with school garden project
Awarded from own funds:-
£100 to 2 pupils travelling to Lourdes 2015
£100 towards Grease Production
Senior Citizen Christmas Lunch 2015 – paid balance
£400 towards school trip to Paris
Supported purchase of school hoodies for 6th year pupils
2 x £20 vouchers at awards ceremony 2015 & 2016
Congratulations to Ben Matovu who won the Power Transformation Challenge.
Powering Transformation is a programme developed by The Transformation Trust, and supported by Dell. This exciting programme gave pupils an opportunity to identify a local issue whether it was in school or in the community and come up with an IT based Solution. The challenge was run for 4 weeks with Ms Hanif and Dell Volunteers assisted. Every pupil participated very well in it and there were many fantastic ideas to solve many issues that affect our everyday lives.
Well done everybody. Great effort.
The Bike Station Academy is a project focused on training young people between the ages of 15 - 21. St Rochs became involved in this project last year with four pupils completing the award. The group are trained in fields such as cycle mechanics and enterprise skills. The Bike Station are aiming to help them set up a community bike hub which can cater for the local area and promote cycling to the people that live there.
Saint Roch's Choir
Bags of Help Initiative
Tesco has teamed up with Groundwork on its Bags of Help initiative across Scotland. The scheme will see three community groups and projects (and we're one of them!) awarded grants of £12,000, £10,000 and £8,000 – all raised from the 5p bag charge. Greenspace Scotland are working with Groundwork to support Scottish communities.
Bags of Help offers community groups and projects in each of Tesco's regions across the UK a share of revenue generated from the 5p charge levied on single-use carrier bags.
The public will now vote in store from 31 October to 13 November on who should receive the £12,000, £10,000 and £8,000 awards. In total, there is over £12.5 million up for grabs.
You can vote for St Roch's in store and the pupils of the school will benefit directly from this initiative.
See the Bags of Help website - www.tesco.com/bagsofhelp
A big thank you to the St. Roch's Secondary Parent Teacher Council who made a successful application to the Trades House of Glasgow Commonwealth Fund. The finance will be used to contribute to the production of 'Grease' which takes place in St. Roch's Secondary School on Wednesday 8th and Thursday 9th June 2016. Cast from the show were presented with the cheque from the Deacon Convener at a special awards ceremony in the Trade Hall in Glasgow. They were accompanied by Ms Simpson and Ms Hewitt from the Parent Teacher Council.
ATTENTION ALL S4 and S5 pupils
Hi All
Just in case you are unaware, the expression of interest Survey Monkey form for next terms college options is live on the Glasgow Online website
Can you please let your pupils know that if they are interested then they should fill out the Level 4-7 Survey on this page
The survey closes at the end of March and we will let you or your pupils know at that time what option they have been successful in gaining a place on in order for them to complete the College participation form
Under-18's St Roch's Panthers Basketball Team, winners of the Glasgow and Surrounding Areas Senior Boys League competition after remaining undefeated throughout the season.
S3 prefects with Archbishop Tartaglia @ Secondary Schools Mass
Congratulations to Kieran McLaughlin of S3. Kieran won the Power Transformation mini challenge working in partnership with Dell by engaging with a problem in his community and inventing a creative technological solution. Power Transformation were very impressed with his idea of oxygen zones to help stop the public's health being damaged by pollution. It was original, professionally presented, and addressed a prominent issue: how pollution has an adverse effect on our communities, not just plant or non-human animal based ecosystems. Well Done Kieran!
S4 Collection for Calais second hand clothes
Tweets by @St_Rochs
Link to Education Privacy Statement
Nationals in a Nutshell
Highers in a Nutshell
SQA 2019 Exam Timetable
SQA Qualifications Guide for pupils and parents
PlanIT Plus
Read our guestbook


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At first, I sat down to update and refresh the look we took in 2012 at celebrities and their Celine Luggage Totes. Once I started digging, though, it became clear that adding a dozen new pictures to the original post wouldn't do stars' love for their Celine bags any justice. Instead, it would require a full follow-up to the original investigation (I use that term loosely), and that's what we have for you today.
Celine Luggage Totes aren't the new thing on the block anymore, and it no longer requires a contact at a high-end boutique and a prepaid waitlist spot to get your hands on one. I still see the bags carried all the time on the streets of New York, though, and celebs of all stripes, from A-List to No List, still trot theirs out regularly. Find out who's been carrying what below.
Alessandra Ambrosio
Jennifer Garner
Kate Walsh
Brooklyn Decker
Kate Upton
Hilary Duff
Kirstie Alley
Bethenny Frankel
Hilary Duff
Gwyneth Paltrow
Tamara Ecclestone
Paulina Rubio
Kylie Jenner
Jordana Brewster
Nicole Kidman
Kate Walsh
Ashley Tisdale
Mischa Barton
Ashley Tisdale
Kylie Jenner
Reese Witherspoon
Lindsay Lohan
Kate Upton
Julianne Hough
Miranda Kerr
Ashley Tisdale
Khloe Kardashian
Hilary Duff
Tamara Ecclestone
Karolina Kurkova
Bethenny Frankel
Khloe Kardashian
Tamara Ecclestone
Lily Aldridge
Nene Leakes
P.S. Please consider supporting our small, bag-loving team by clicking our links before shopping or checking out at your favorite online retailers like Amazon, Neiman Marcus, Nordstrom, or any of the listed partners on our shop page. We truly appreciate your support!


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Crude steel production in the 66 countries reporting to the World Steel Association totalled 114 million tonnes in November 2010, 5.1% higher than in November 2009. However, the eleven months total was 1,281 million tonnes, 16.2% higher than the same period last year.
Excluding China, the monthly rise was 5.3%, whereas the year to date total increased by 21.7%. All regions showed an increase in the month and year to date, except for the November total for South America and Oceania.
Steel production in the 27 member countries of the European Union increased by 4.1% in November 2010 compared to the previous November to 14.7 million tonnes; the eleven months total, however, was 26% higher at 160 million tonnes. German production rose in November by 8%, bringing the eleven months total up 37% to 40.6 million tonnes. Production in Italy, on the other hand, increased by 19% in November, while the year to date total rose by 30% to 23.8 million tonnes. However, Spanish steel production actually fell by 20% in the month, although the year to date total increased by 13.6% to 15.1 million tonnes. French November production was down by 1.6%, while the eleven months total rose by 21.7% to 14.3 million tonnes. UK monthly steel production dropped by 22.8%, the largest fall in the EU, bringing the year to date total up 1.6% to 9.1 million tonnes. The Belgian eleven month total rose by nearly 45% to 7.4 million tonnes.
In the rest of Europe the largest producer is Turkey, where steel production increased by 15.7% in November, and by 12.9% in the year to date to 26.1 million tonnes. Although the monthly total in Serbia actually fell compared to November 2009, the eleven months total rose by 31.4% to 1.2 million tonnes, just ahead of the Swiss total.
While Turkish steel production rose by 13% in 2010 compared to 2009, Turkish steel exports fell by 10% to 14.5 million tonnes. The largest drop in exports was in deformed reinforcing bars, down from 8.1 to 5.4 million tonnes in 2010. However, exports of billets rose from 2.2 to 3.4 million tonnes. Total imports of steel in the eleven months rose by 8.7% to just under 10 million tonnes.
Turkey's largest export market was the Middle East which took 6.9 million tonnes (47.5%) in the first eleven months of 2010; United Arab Emirates, Saudi Arabia and Iraq were the three largest markets within that. Africa accounted for 2.8 million tonnes (19.5%) in 2010 with Egypt, Libya and Algeria the top three markets. The European Union only accounted for 1.8 million tonnes (12.7%) with Italy, Romania and the United Kingdom being the largest recipients. Asian countries accounted for less than one million tonnes (6.3%) with Singapore and Thailand taking over 60% of the tonnage.
According to ACEA, the European car manufacturers' association, European car registrations fell by 6.5% in November, and were down by 5.1% in the eleven months. Registrations in Germany, the largest market, decreased by 6.2% in November, and by 25% in the year to date to 2.7 million cars. French registrations were down by 10.8% in November but only by 2.4% in the year total to 2.0 million cars. The UK total showed an 11.5% fall in November, but the eleven months total was 3.4% up at 1.9 million cars, while in Italy monthly registrations were down by 21% with the year 8% down at 1.8 million cars. The fifth largest market, Spain, was 25% down in November, but 6% up in the year to date at 913 thousand cars.
In the CIS countries crude steel production in Russia rose by 2.7% in November bringing the year to date total up 12% to 61.1 million tonnes. Ukrainian November production increased by 6.8%, making the eleven months total 30 million tonnes, an increase of 12.7%. Steel production in Kazakhstan only increased by 3.3% in the year to date to 3.9 million tonnes.
Steel production in the USA rose by 13% in November, bringing the year to date total up 41% to 73.9 million tonnes. Canadian production was up just 2% in November, while the eleven months total was up by 44% to 12 million tonnes. Mexican steel production rose by 7.7% in the month, although the year to date total increased by 21% to 15.3 million tonnes.
In South America, Brazilian crude steel production actually fell by 2.8% in November, although the year to date total was up 27% to 30.4 million tonnes compared to last year. Argentinian production in November rose by 10%, bringing the eleven months total up nearly 31% to 4.7 million tonnes. Venezuelan crude steel production, on the other hand, decreased by 37% in the month, while the year to date total was down by 45% to 1.9 million tonnes.
In Africa and the Middle East steel production rose overall. Iranian production jumped by 23% in November, while the year to date total rose by 7.8% to 10.9 million tonnes. The South African monthly total was only 3.7% up with the eleven months total 14% higher at 7.75 million tonnes. In Egypt steel production rose by nearly 26% in the month, and was 16.5% up in the year to date to 5.8 million tonnes. Saudi Arabian production rose by 8.6% in the month bringing the January to November total up 8.3% to 4.6 million tonnes. Qatar steel production increased by 39% in the eleven months to 1.8 million tonnes.
In Asia Chinese steel production only increased by 4.8% in November, while the eleven months total was up by 10% to 577 million tonnes, 45% of the world total. Indian steel production increased by 3.2% in the month, and by 7.4% in the year to date to 61.3 million tonnes. Japanese production showed a very slight increase in November, while the year to date total was up by nearly 28% to 100 million tonnes. South Korean steel production rose by 16.4% in the month, and by 19.2% in the year to date to 52.5 million tonnes. Taiwanese steel production, however, fell by 12% in November, although the January to November total increased by 25.6% to 17.8 million tonnes.
Phil Hunt, ISSB Ltd
|Oct 2009
|Jan 2010
|Apr 2010
|Jul 2010
|Oct 2010
|Nov 2009
|Feb 2010
|May 2010
|Aug 2010
|Nov 2010
|Dec 2009
|Mar 2010
|Jun 2010
|Sep 2010
|Dec 2010


text: This is a sponsored post in collaboration with Clorox. All thoughts are my own. When I was a kid, birthdays were fun, exciting, something I was anxiously waiting for for months. The party, the presents….the cake! As a young adult, my birthday was a cause for days long celebrations, nights of dancing and flirting, and days of dinners, outings and shopping for trendy clothes with friends. Those … [Read more...]


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Thursday, April 24, 2008
My New Name For A Blog
What Jane Said About What Digby Said.
Maybe it's nothing more than the fact that it's been sunny and warm, with the daylight extending almost to 8:00 pm. Maybe it's the lilacs and woad in full bloom in my back yard (For the last few days, I swear that I've literally been able to hear the Earth thrumming: Beltane, Beltane, Beltane!) Maybe it's the win Hillary pulled off in Pennsylvania due, in large part, to those old women in boxy blazers. Or maybe there's more, but, in any case, I'm lately feeling quite hopeful about almost everything.
As Jane notes that Digby said:
To me, this primary is actually a good thing for the fall. All this hand wringing strikes me as typical Democratic nervous nellie-ism. A huge increase in Democratic voter registration, building of strong ground operations in most states, new technologies being beta tested, lots of media coverage and battle testing for the nominee are of benefit to the nominee in the fall. Meanwhile, the Democrats stay at center stage while McCain wanders around in obscurity, failing to raise money and leaving a trail of gaffes in his wake. As long as they don't know at whom to aim their fire the Republicans can't cement their narrative. In the end, I remain convinced that we are going into an election that is so fundamentally seismic that either of them can win it, even if more closely than we might want, due to the breakthrough nature of their campaigns. The primary continuing on is not going to change that.
I don't think people realize that the democratization the internet has brought to the system is also one of the main reasons why the campaign goes on. If you think superdelegates are undemocratic, back in the bad old days (of a couple of cycles ago) big party donors pulled the strings by pulling the money when they decided that someone had no chance to win. Today, both candidacies are where they are on the basis of avid small donor supporters contributing online and that's prolonged things past the point where it would have in the past. Thousands of Clinton supporters keep sending her money-- ten million since last night, apparently. So, if you don't like the fact that the campaign continues, blame the internet. It wouldn't have happened under the old paradigm.
Obama supporters could do themselves and the party a big favor if they'd quit crying about how that eeeeevvvvvvvvvviiiiiiillllllll woman needs to shut up "so we can speak with one (man's) voice!" and step down before she "destroys" the guy who is supposedly going to be able to stand up to the Republican slime machine. I didn't like it when Sandra Day and her Gang of Four shut down democracy in Florida and I don't like it when Obama's supporters try to do it in the Democratic primary. I am just saying. Have a little faith -- in your candidate and in the process.
As Digby explains:
the fact that Clinton is still winning big primaries and getting campaign contributions makes it ridiculous to expect her --- or any politician --- to quit (no matter what the NY Times editorial board says.) She has a legitimate constituency (nearly half the voters) in the party that wants her to see this through. As Somerby says in response to Maureen Dowd's typically daft (and equal opportunity insulting) column this morning:
This year's campaign has shown what can happen when a party has two closely-matched candidates. There are potential downsides for the party, as anyone can see. But journos like Dowd think it's their role to demand that the person they hate should just quit. Those million-plus Democrats [who voted for her yesterday] don't exist in Dowd's world. In Dowd's world, Dowd wants Clinton to quit. And so, by the laws of childish dreams, "the Democrats" must want that too.
Since I don't think the Democratic Party will crumble from the stress of finishing up the primaries (or even a brokered convention for that matter) until Obama officially wraps up the number of delegates and superdelegates to go over the magic number, I think she has a right to continue.
4 comments:
- ifthethunderdontgetya™³²®© said...
Hillary supporters could do the party a big favor if they'd drop the "Obamabot" namecalling.
Also, I'd appreciate it if they'd stop running Karl Rove's campaign for him.
Hecate, this is for you. Enjoy. :)
- LittleIsis said...
You're right Hecate!!! (when aren't you?) We Dems should stop freaking out. She's got more Democratic primary votes than any candidate in history (more than FDR!!!) Give me a break. And as long as she mantains her new popular vote lead, I think she might win at the convention. And ifthethunderdontgetya, I understand what you mean, but Obama supporters should stop harassing her supporters and calling them racists and acting like sexists pigs, and trying to force her out of the race.
The "You get out of the race" "No, YOU get out opf the race" is ridiculous. This is called democracy, and it's playing out exactly as it should when the system works. The voters in all the states are having their say, something we haven't seen in way too long. It may be nerve-wracking for some of us, and that's just too bad. All this means is that we have two superb candidates and excellent chances of winning it all in November. Not exactly something to lament.


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Fucked Up will be playing a one off show to support The Vaccines at London's Alexandra Palace on November 17th.
Will The Vaccines be able to handle our Canadian bruisers? Will they escape without being upstaged? Perhaps so, check out their take on the Minor Threat cover of the The Standells' track "Sometimes Good Guys Don't Wear White." They're even joined on stage by original 'Threat member Lyle Preslar.


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Big Readr update!
We're pleased to announce that a major Readr update went live on the App Store today. The improvements in this version include a simplified process for importing recipes and a new icon. We hope these enhancements will make you enjoy using Readr even more!
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