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Romidepsin (FK228) improves the survival of allogeneic skin grafts through downregulating the production of donor-specific antibody via suppressing the IRE1α-XBP1 pathway / 浙江大学学报(英文版)(B辑:生物医学和生物技术)
Yuliang GUO; Siyu SONG; Xiaoxiao DU; Li TIAN; Man ZHANG; Hongmin ZHOU; Zhonghua-Klaus CHEN; Sheng CHANG.
Journal of Zhejiang University. Science. B ; (12): 392-406, 2022.
Article in English | WPRIM | ID: wpr-929069
Antibody-mediated rejection (AMR) is one of the major causes of graft loss after transplantation. Recently, the regulation of B cell differentiation and the prevention of donor-specific antibody (DSA) production have gained increased attention in transplant research. Herein, we established a secondary allogeneic in vivo skin transplant model to study the effects of romidepsin (FK228) on DSA. The survival of grafted skins was monitored daily. The serum levels of DSA and the number of relevant immunocytes in the recipient spleens were evaluated by flow cytometry. Then, we isolated and purified B cells from B6 mouse spleens in vitro by magnetic bead sorting. The B cells were cultured with interleukin-4 (IL-4) and anti-clusters of differentiation 40 (CD40) antibody with or without FK228 treatment. The immunoglobulin G1 (IgG1) and IgM levels in the supernatant were evaluated by enzyme-linked immunosorbent assay (ELISA). Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and western blotting were conducted to determine the corresponding levels of messenger RNA (mRNA) and protein expression in cultured cells and the recipient spleens. The results showed that FK228 significantly improved the survival of allogeneic skin grafts. Moreover, FK228 inhibited DSA production in the serum along with the suppression of histone deacetylase 1 (HADC1) and HDAC2 and the upregulation of the acetylation of histones H2A and H3. It also inhibited the differentiation of B cells to plasma cells, decreased the transcription of positive regulatory domain-containing 1 (Prdm1) and X-box-binding protein 1 (Xbp1), and decreased the expression of phosphorylated inositol-requiring enzyme 1 α (p-IRE1α), XBP1, and B lymphocyte-induced maturation protein-1 (Blimp-1). In conclusion, FK228 could decrease the production of antibodies by B cells via inhibition of the IRE1α-XBP1 signaling pathway. Thus, FK228 is considered as a promising therapeutic agent for the clinical treatment of AMR.
Animals , Depsipeptides , Endoribonucleases , Hematopoietic Stem Cell Transplantation , Histone Deacetylase Inhibitors/pharmacology , Mice , Protein Serine-Threonine Kinases , Skin Transplantation
Histone Deacetylase Inhibitors in the in Vitro Expansion of Hematopoietic Stem Cells / 中国医学科学院学报
Miao CHEN; Jun-Ling ZHUANG.
Acta Academiae Medicinae Sinicae ; (6): 109-115, 2021.
Article in Chinese | WPRIM | ID: wpr-878706
The self-renewal and differentiation of hematopoietic stem cells(HSCs)are highly regulated by epigenetic modification,in which histone acetylation can activate or silence gene transcription.Histone deacetylase inhibitors(HDACIs)can inhibit the activity of histone deacetylase in HSCs to increase histone acetylation.A variety of HDACIs,such as trichostatin A and valproic acid,are used to expand HSCs in vitro,especially cord blood HSCs,combined with cytokines in serum-free culture to obtain more long-term repopulating cells.HDACIs promote the transcription of pluripotent genes related to stem cell self-renewal and inhibit the expression of genes related to differentiation,so as to promote the expansion and inhibit differentiation of HSCs.The expansion of cord blood HSCs by small molecular HDACIs in vitro is expected to improve the quantity of cord blood HSCs.The further research will focus on high-throughput screening for the most powerful HDACIs and the highly selective HDACIs,exploring the combination of epigenetic modifiers of different pathways.
Epigenesis, Genetic , Fetal Blood , Hematopoietic Stem Cells , Histone Deacetylase Inhibitors/pharmacology , Valproic Acid/pharmacology
Effects of brain-specific knockout of Sirt6 gene on cerebral cortex in aged mice / 中华行为医学与脑科学杂志
Xinhuan WANG; Lin HOU; Bin YIN; Wei LIU; Xiaozhong PENG.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 481-488, 2021.
Objective:To study the effect of senescence gene silent information regulator 6 (Sirt6) knockout on the brain of aged mice.Methods:Sirt6-flox transgenic mice were constructed, and the mouse brain tissue was specifically knocked out by Emx1-Cre tool mice.According to genotyping, 11 wild-type mice were selected as control group(WT group) and 10 Sirt6 gene konckout mice were selected as conditional knockout group(cKO group). Body size and body weight of the aged mice were measured and cerebral cortex thickness was measured by HE staining.Brain neurogenesis was analyzed with EdU markers.The expression of RNA-binding protein HuR and apoptosis-related protein Caspase-3 were detected by Western blot.Meanwhile, histone acetylation levels in the cortex were detected.Results:Sirt6 brain tissue-specific knocked out mice were successfully constructed.Compared with the brain tissue area((2.07±0.22) cm 2)and cortical thickness ((970.56±80.91) μm) of WT mice in the 12-month-old group, the brain tissue area ((1.61±0.14)cm 2) and cortical thickness ((822.88±53.94) μm) in Sirt6 cKO group were smaller, and the differences were statistically significant (both P<0.05). EdU incorporation into nerve cells showed that the number of EdU incorporation into periventricular nerve cells in cKO group was lower ((4.75±1.48)) than that in WT group ((10.29±1.93)). The difference was statistically significant ( P<0.001). In the experiment of 17 months age group, mice in cKO group were smaller in body size, lower in body weight ((29.00±1.08) g) and smaller in brain area ((1.54±0.55)cm 2)compared with WT group in body size, body weight ((35.25±4.17) g) and brain tissue area ((1.98±0.18) cm 2)(both P<0.05). The expression of Caspase-3 and HuR in cortical proteins of these two age groups decreased( t=2.95, 5.38, both P<0.05), and the expression of H3K9ac and H3K56ac increased( t=3.53, 2.78, both P<0.05), but the expression of Sirt1 homologous gene remained unchanged( t=1.26, P>0.05). Conclusion:The specific deletion of Sirt6 in brain tissue can lead to the decrease of brain neurogenesis in aged mice, and the aggravation of aging and the increase of apoptosis, which may be the reason for the thinning of cerebral cortex and brain tissue atrophy.The molecular mechanism is speculated to be related to the increase of acetylation level after Sirt6 knockout
Cellular environment controls the dynamics of histone H3 lysine 56 acetylation in response to DNA damage in mammalian cells
Vadla, Raghavendra; Chatterjee, Nirupama; Haldar, Devyani.
J Biosci ; 2020 Jan; : 1-12
Article | IMSEAR | ID: sea-214347
Epigenetic changes play a crucial role in sensing signals and responding to fluctuations in the extracellularenvironment. How the cellular micro-environment affects DNA damage response signalling in chromatincontext is not extensively studied. Histone acetylation is dynamic and very sensitive to changes in theextracellular environment. Existing literature on H3 lysine 56 acetylation (H3K56ac) levels upon DNA damagein mammals presents a conflicting picture. The occurrence of both increased and decreased H3K56ac uponDNA damage in our experiments led us to investigate the role of the micro-environment on H3K56ac. Here,we show that the global levels of H3K56ac increase as cells grow from low density to high density whileSIRT1 and SIRT6 expression decrease. Additionally, rising lactic acid levels increase H3K56ac. Our resultsshow that cell density and accumulation of metabolites affect dynamics of H3K56ac in response to DNAdamage. Upon DNA damage, H3K56ac increases in low density cells with low initial acetylation, whileacetylation decreases in high cell density cells. These results highlight that H3K56ac levels upon DNA damageare dependent on the metabolites in the extracellular milieu which impact chromatin structure by regulatingchromatin modifying enzymes. Accumulation of lactic acid at high cell density reflects conditions similar to thetumour micro-environment. As H3K56ac increases in tumours, lactic acid and low pH might alter H3K56ac intumours, leading to deregulated gene expression, contributing to tumour progression.
Role of histone acetylation in the pathogenesis of IgA nephropathy / 上海交通大学学报(医学版)
Qin DAI.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 1063-1068, 2020.
Objective: To study the role of abnormal histone acetylation modification in the pathogenesis of IgA nephropathy (IgAN), and explore the pathogenesis of IgAN from the perspective of epigenetics. Methods: 30 patients with primary IgAN and 19 healthy controls were included. Of the 19 controls, 16 cases were used to collect blood samples and 3 cases were used to collect normal renal tissues. Peripheral venous blood was collected to extract mononuclear cells (PBMC) for detection. The levels of histone H3 and H4 acetylation in PBMC of IgAN group and healthy control group were detected by ELISA. Real-time PCR was used to detect the mRNA expressions of P300, CREBBP, HDAC1-3, HDAC7, HDAC8, C1GALT1 and ST6GALNAC2. The acetylation levels of histone H3 and H4 in promoter regions of C1GALT1 and ST6GALNAC2 were detected by chromatin immuneprecipitation (CHIP). The protein expression of HDAC1 and H3Ac was detected by immunofluorescence. T-test was used for statistical analysis. Results: Compared with the healthy control group, the acetylation levels of H3 and H4 in IgAN group were significantly higher than those in healthy control group (P=0.035, P=0.012);the mRNA expressions of P300, CREBBP, HDAC1, HDAC8 and ST6GALNAC2 were significantly increased in IgAN group (P=0.002, P=0.001, P=0.001, P=0.045 P=0.012); the mRNA expressions of HDAC2 and C1GALT1 were decreased significantly (P=0.035, P=0.008); the acetylation degree of H3 and H4 in C1GALT1 promoter region was significantly decreased (P=0.043, P=0.005), the degree of H3 and H4 acetylation in the promoter region of ST6GALNAC2 gene was increased significantly in IgAN group (P=0.038, P=0.021);immune fluorescent results of renal tissue indicated that the HDAC1 protein expression was increased obviously, and H3Ac protein expression was significantly reduced in the renal tissue of IgAN patiegts. Conclusion: Abnormal acetylation modification exists in IgAN patients. Histone acetylation modification may participate in the pathogenesis of IgAN by regulating the expression of glycosylase gene.
Enriched environment improves sevoflurane-induced cognitive impairment during late-pregnancy via hippocampal histone acetylation
Yu, Zhiqiang; Wang, Jinxin; Zhang, Peijun; Wang, Jianbo; Cui, Jian; Wang, Haiyun.
Braz. j. med. biol. res ; 53(10): e9861, 2020. graf
Article in English | LILACS, ColecionaSUS | ID: biblio-1132476
Fetal exposure to sevoflurane induces long-term cognitive impairment. Histone acetylation regulates the transcription of genes involved in memory formation. We investigated whether sevoflurane exposure during late-pregnancy induces neurocognitive impairment in offspring, and if this is related to histone acetylation dysfunction. We determined whether the effects could be reversed by an enriched environment (EE). Pregnant rats were exposed to 2.5% sevoflurane or control for 1, 3, or 6 h on gestational day 18 (G18). Sevoflurane reduced brain-derived neurotrophic factor (BDNF), acetyl histone H3 (Ac-H3), and Ac-H4 levels and increased histone deacetylases-2 (HDAC2) and HDAC3 levels in the hippocampus of the offspring on postnatal day 1 (P1) and P35. Long-term potentiation was inhibited, and spatial learning and memory were impaired in the 6-h sevoflurane group at P35. EE alleviated sevoflurane-induced cognitive dysfunction and increased hippocampal BDNF, Ac-H3, and Ac-H4. Exposure to 2.5% sevoflurane for 3 h during late-pregnancy decreased hippocampal BDNF, Ac-H3, and Ac-H4 in the offspring but had no effect on cognitive function. However, when the exposure time was 6 h, impaired spatial learning and memory were linked to reduced BDNF, Ac-H3, and Ac-H4, which could be reversed by EE.
Animals , Female , Pregnancy , Rats , Cognitive Dysfunction , Acetylation , Histones , Maze Learning , Brain-Derived Neurotrophic Factor , Sevoflurane , Hippocampus
Molecular cloning and subcellular localization of six HDACs and their roles in response to salt and drought stress in kenaf (Hibiscus cannabinus L)
Wei, Fan; Tang, Danfeng; Li, Zengqiang; Kashif, Muhammad Haneef; Khan, Aziz; Lu, Hai; Jia, Ruixing; Chen, Peng.
Biol. Res ; 52: 20, 2019. tab, graf
Article in English | LILACS | ID: biblio-1011422
BACKGROUND: Histone acetylation is an important epigenetic modification that regulates gene activity in response to stress. Histone acetylation levels are reversibly regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). The imperative roles of HDACs in gene transcription, transcriptional regulation, growth and responses to stressful environment have been widely investigated in Arabidopsis. However, data regarding HDACs in kenaf crop has not been disclosed yet. RESULTS: In this study, six HDACs genes (HcHDA2, HcHDA6, HcHDA8, HcHDA9, HcHDA19, and HcSRT2) were isolated and characterized. Phylogenetic tree revealed that these HcHDACs shared high degree of sequence homology with those of Gossypium arboreum. Subcellular localization analysis showed that GFP-tagged HcHDA2 and HcHDA8 were predominantly localized in the nucleus, HcHDA6 and HcHDA19 in nucleus and cytosol. The HcHDA9 was found in both nucleus and plasma membranes. Real-time quantitative PCR showed that the six HcHDACs genes were expressed with distinct expression patterns across plant tissues. Furthermore, we determined differential accumulation of HcHDACs transcripts under salt and drought treatments, indicating that these enzymes may participate in the biological process under stress in kenaf. Finally, we showed that the levels of histone H3 and H4 acetylation were modulated by salt and drought stress in kenaf. CONCLUSIONS: We have isolated and characterized six HDACs genes from kenaf. These data showed that HDACs are imperative players for growth and development as well abiotic stress responses in kenaf.
Stress, Physiological/physiology , Hibiscus/enzymology , Histone Acetyltransferases/physiology , Droughts , Histone Deacetylases/physiology , Transcriptional Activation/physiology , Cloning, Molecular , Hibiscus/growth & development , Hibiscus/physiology , Real-Time Polymerase Chain Reaction
Regulation on brain-derived neurotrophic factor and relevant mechanism of anemarrhena saponin in H2O2-induced SH-SY5Y cells / 上海交通大学学报(医学版)
Shuang-Shuang YANG.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 578-585, 2019.
Objective • To investigate the effect of anemarrhena saponin (ZMS) on mRNA level of brain-derived neurotrophic factor (BDNF) and relevant mechanism in oxidative stress damage of SH-SY5Y cells. Methods • SH-SY5Y cells treated with H2O2 were chosen as cell models of oxidative stress. Cell viability was determined using cell counting kit-8 (CCK-8). The mRNA levels of BDNF and its important transcripts were detected by quantitative real-time PCR (qPCR). The histone deacetylases (HDACs) activity fluorescence quantification assay kit was used to measure the effect of ZMS on HDACs activity. Western blotting was used to detect the protein expression levels of acetylated histone H3, acetylated histone H4, specific acetylation site-related proteins, and HDAC1/2/3. Results • qPCR showed that ZMS could increase the mRNA levels of BDNF and its transcript in the cell models. Western blotting showed that ZMS pretreatment could increase the protein levels of acetylated histone H3, acetylated histone H4 and acetylated histone H3K14, and there was no significant effect on protein levels of HDAC1/2/3. In addition, HDACs activity fluorescence quantification assay kit showed that ZMS could inhibit HDACs activity significantly. Conclusion • ZMS can increase the mRNA levels of BDNF and its transcript in oxidative stress damage cell models, which may be related to the regulation of histone acetylation level
Effect of Long-time Slow-frozen Storage on Epigenetic Changes of Histone in Cleavage-stage Embryos / 中山大学学报(医学科学版)
Jia-xuan MA; Bing CAI; Yan-wen XU; Can-quan ZHOU.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 781-787, 2019.
@#【Objective】To investigate the impact of long- term storage time on epigenetic modification of histone in human cleavage stage embryos.【Methods】According to the length of storage time,donated embryos after slow-freezing were divided into 3 groups :6-year group ,9-year group ,and 12-year group ,while the control group consisted of donated fresh embryos. Immunocytochemistry was performed to compare the expression levels of HDAC1, H3K9ac, H3K4me3 ,and H3K9me3 among 4 groups. Transcription levels of HDAC1 ,SUV39H1 ,SETDB1 ,and KDM5A were analyzed through Single-Cell qRT-PCR.【Results】The relative abundances of HDAC1 and SUV39H1 mRNA showed no significant differences among 4 groups(P > 0.05). SETDB1 exhibited a climbing pattern as storage time increased,but no significant difference was observed(P > 0.05). There were no differences in H3K9 trimethylation and H3K9 methylation among 4 groups. However ,the expression level of KDM5A increased with the increasing storage time(P < 0.05).【Conclusions】 Storage time did not affect the expression of deacetylase HDAC1,methylase SUV39H1 and SETDB1. H3K9ac/me3 and H3K4me3 also exhibited no significant difference as the storage time increases. However ,the increasing storage length might induce the elevating expression of demethylase KDM5A,which may be associated with inhibition of embryonic transcription.
Effect of histone acetylation on osteogenic differentiation of periodontal ligament stem cells derived from periodontitis tissue / 华西口腔医学杂志
Jin SUN; Yun LIU; Qian QU; Juan QU; Wei LUO; Feng ZHANG; Min WU.
West China Journal of Stomatology ; (6): 102-105, 2019.
Epigenetics is defined as a change in gene expression without the alteration of the genetic sequence. Such a change would be inherited by offspring. Histone acetylation is a type of epigenetics. Existing studies proposed that chronic periodontitis is related to epigenetic modification. In this review, we summarised the influence of chronic periodontitis on periodontal ligament stem cells by histone acetylation.
Acetylation , Cell Differentiation , Cells, Cultured , Histones , Metabolism , Osteogenesis , Periodontal Ligament , Stem Cells , Physiology
Insights on the epigenetic mechanisms underlying pulmonary arterial hypertension
Luna, R C P; de Oliveira, Y; Lisboa, J V C; Chaves, T R; de Araújo, T A M; de Sousa, E E; Miranda Neto, M; Pirola, L; Braga, V A; de Brito Alves, J L.
Braz. j. med. biol. res ; 51(12): e7437, 2018. tab, graf
Article in English | LILACS | ID: biblio-974252
Pulmonary arterial hypertension (PAH), characterized by localized increased arterial blood pressure in the lungs, is a slow developing long-term disease that can be fatal. PAH is characterized by inflammation, vascular tone imbalance, pathological pulmonary vascular remodeling, and right-sided heart failure. Current treatments for PAH are palliative and development of new therapies is necessary. Recent and relevant studies have demonstrated that epigenetic processes may exert key influences on the pathogenesis of PAH and may be promising therapeutic targets in the prevention and/or cure of this condition. The aim of the present mini-review is to summarize the occurrence of epigenetic-based mechanisms in the context of PAH physiopathology, focusing on the roles of DNA methylation, histone post-translational modifications and non-coding RNAs. We also discuss the potential of epigenetic-based therapies for PAH.
Humans , DNA Methylation/genetics , RNA, Untranslated/genetics , Epigenesis, Genetic/genetics , Histone Code/genetics , Hypertension, Pulmonary/physiopathology , Hypertension, Pulmonary/genetics , Pulmonary Artery/pathology , Down-Regulation/genetics , Up-Regulation/genetics , Ubiquitination/genetics , Molecular Targeted Therapy , Hypertension, Pulmonary/therapy
Regulatory mechanism of ginsenoside Rd on histone H3 acetylation / 中草药
Jia-Shu ZHANG.
Chinese Traditional and Herbal Drugs ; (24): 2931-2936, 2018.
Objective Based on quantitative proteomics analysis and molecular biology experimental verification, the regulatory mechanism of ginsenoside Rd on histone H3 acetylation levels was elucidated. Methods The effects of ginsenoside Rd on the dynamic changes of proteome of HEK293T cells were detected by.stable isotope labeling with amino acid (SILAC) technique and LC-MS/MS; Quantitative proteomics database analysis was used to monitor the changes in histone acetyltransferase HATs and histone deacetylase HDACs expression levels. Western blotting and qRT-PCR were used to verify the changes of related protein expression and transcriptional level. Gene knockdown experiments were performed using siRNAs to determine the role of ginsenoside Rd in regulating the level of acetyl modifications at histone H3K9 and K18 sites. Results The histone H3K9ac, H3K18ac expression levels in HEK293T cells decreased after ginsenoside Rd treatment, but the P300 catalytic modification of these two sites did not change significantly; At the same time, ginsenoside Rd up-regulated the transcription and expression of HDAC2, and siHDAC2 treatment reversed the down-regulation effects of ginsenoside Rd on H3K9ac and H3K18ac in HEK293T cells. Conclusion Ginsenoside Rd down-regulates the acetylation level of lysine at histone H3K9 and K18 sites by up-regulating HDAC2, thereby affecting transcriptional activation of downstream genes.
The effect of sevoflurane anesthesia on learning and memory and hippocampal histone acetylation in mice / 中华行为医学与脑科学杂志
Dengxin ZHANG; Bingxin XUE; Bin ZHANG; Gaoshang CHAI.
Objective To investigate the effect of sevoflurane anesthesia on learning and memory in mice and the relationship with histone acetylation. Methods Thirty-six adult male C57BL/6 mice were ran-domly divided into control group(mice inhaled 95% O2 for 6 h), sevoflurane (Sevo) group (1. 5% Sevo group, 2% Sevo group, 3% Sevo group:mice inhaled 1. 5%, 2% and 3% sevoflurane for 6 h respectively) , sevoflurane + SAHA (Sevo + SAHA) group (mice were intraperitoneally injected with histone deacetylase inhibitor SAHA ( 25 mg/kg) . And 1 h later, 3% Sevo was inhaled continuously for 6 hours. ) and SAHA group(mice were intraperitoneally injected with histone deacetylase inhibitor SAHA (25 mg/kg)). The abil-ity of learning and memory in mice was estimated by Morris water maze. The expression levels of Ac-H3, BDNF and Syt-I protein in the hippocampus were detected by Western blot. Results In Morris water maze, 3% sevoflurane anesthesia significantly prolonged the escape latency((46. 91±1. 84)s),and significantly decreased the ratio of target time((35. 84±5. 40)%) compared with that of control group((23. 46±2. 67)s, (49. 74±4. 91)%,P<0. 05). Compared with 3% Sevo group,the ratio of target time in Sevot+SAHA group ((46. 86±4. 37)%) was increased(P<0. 05). Moreover,3% sevoflurane anesthesia significantly decreased the expression levels of Ac-H3 (10. 23±2. 45),BDNF (6. 72±1. 21) and Syt-I (8. 25±2. 11) in the hippo-campus compared with that of control group((15. 45±2. 58),(10. 17±1. 45) and (15. 02±3. 38),P<0. 05) . However,pre-administration of the histone deacetylase inhibitor SAHA significantly increased the ra-tio of target time in Morris water maze,and improved the expression levels of Ac-H3 (14. 06±2. 79),BDNF (10. 13±1. 06) and Syt-I (14. 16±3. 66) in Sevo+SAHA group compared with that of Sevo group (P<0. 05) . Conclusion The high dose of sevoflurane anesthesia can induce learning and memory impairment through the inhibitation of histone acetylation in the hippocampus.
Botulinum neurotoxin serotype A heavy chain intervenes in the H3 acetylation: A preliminary study / 中国比较医学杂志
Jing LAN; Fu LIU; Yang GAO; Zhiqiang LI; Ya LIU; Mingwei ZHAO; Haixia YUAN; Xiaqing LI.
Chinese Journal of Comparative Medicine ; (6): 4-9, 2018.
Objective To investigate the effect and molecular mechanism of botulinum neurotoxin serotype A (BoNT/A) heavy chain on neuron regeneration. Methods Cell culture, rats, immunofluorescence, SDS-PAGE and western blot, etc. were adopted in this study to explore the alterations of histone-3 acetylation (acetyl-H3 ) by local treatment of BoNT/A heavy chain to spinal cord injury (SCI) in rats (in vivo) or by adding it into cell culture (in vitro). Meanwhile, the relevance of acetyl-H3 to neurite out-growth based on SCI and cell culture with BoNT/A heavy chain application was approached as well. Results The application of BoNT/A heavy chain to cultured Neuro-2a cells increased the level of H3 acetylation. The increase of H3 acetylation was paralleled with the growth of neuritogenesis. Also, the neuronal treatment of BoNT/A heavy chain to SCI promoted the re-growth of neuronal processes surrounding the lesions. The growth of neuronal processes was positively correlated to the level of H3 acetylation. During the periods of BoNT/A heavy chain treatment in vivo or in vitro, the increase of H3 acetylation showed two peaks. Conclusions BoNT/A heavy chain increased the H3 acetylation, which might be one of its neuritogenic mechanisms.
The effects of sodium fluoride on histone acetylation of CyclinD1/cyclin-dependent kinases 4 gene in human osteoblasts / 中华地方病学杂志
Weimin YAN; Juan MING; Tongzhao YOU; Xilan WANG; Peng LUO; Xueli PAN.
Chinese Journal of Endemiology ; (12): 13-18, 2018.
Objective To establish an model of fluorosis with human primary osteoblasts in vitro and to detect the influences of different doses of sodium fluoride (NaF) on histone acetylation of CyclinD1,cyclindependent kinases 4 (CDK4) gene in human osteoblasts,then to explore the molecular mechanism of skeletal fluorosis from epigenetic perspective of the cell cycle regulation related genes.Methods Human primary osteoblasts from bone tissues of trauma surgery healthy people (car accident) were isolated by enzyme digestion and identified.The osteoblasts were treated with 0,125,250,500 and 1 000 μmol/L NaF for 72 h.The level of histone acetylation (H3K9,H3K14,H4K12,H4K16) in the transcription regulatory region (ChIP1 region) and in the coding region (ChIP2 region) of CyclinD1 and CDK4 genes were detected by quantitative chmmatin immuno-precipitation (Q-ChIP).Results ①After human osteoblasts were treated with 0,125,250,500 and 1 000 μmol/L NaF,respectively,the levels of histone acetylation of H3K9 in ChIP1 transcription regulatory region of CyclinD1 gene were 1.152 ± 0.104,1.174 ± 0.187,1.090 ± 0.176,1.170 ± 0.197 and 1.147 ± 0.097,respectively,the differences were not statistically significant (F =0.524,P > 0.05);the average levels of histone acetylation of H3K14 were 1.495 ± 0.117,1.465 ± 0.069,1.470 ± 0.187,1.760 ± 1.089 and 1.341 ± 0.443,the differences were not statistically significant (F =0.841,P > 0.05);the levels of histone acetylation of H4K12 were 1.239 ± 0.286,0.702 ± 0.063,0.765 ± 0.370,1.011 ± 0.321 and 1.319 ± 0.026,the differences were not statistically significant (F =2.329,P > 0.05);the levels of histone acetylation of H4K16 were 1.452 ± 0.217,1.621 ± 0.165,1.462 ±0.090,1.510 ± 0.146 and 1.564 ± 0.154,the differences were not statistically significant (F =0.123,P > 0.05).②The levels of histone acetylation of H3K9 in ChIP1 transcription regulatory region of CDK4 were 1.472 ± 0.163,1.580 ± 0.161,1.585 ± 0.132,1.451 ± 0.136 and 1.560 ± 0.039,the differences were not statistically significant (F =0.461,P > 0.05);the levels of histone acetylation of H3K14 were 0.919 ± 0.149,0.900 ± 0.059,0.911 ±0.162,0.663 ± 0.049 and 0.841 ± 0.122,the differences were not statistically significant (F =0.974,P > 0.05);the levels of histone acetylation of H4K12 were 0.456 ± 0.142,0.911 ± 0.126,0.969 ± 0.185,1.110 ± 0.146 and 0.931 ± 0.141,the differences were not statistically significant (F=5.459,P > 0.05);the levels of histone acetylation of H4K16 were 1.315 ± 0.083,1.374 ± 0.153,1.423 ± 0.055,1.300 ± 0.132 and 1.385 ± 0.696,the differences were not statistically significant (F =1.663,P > 0.05).③The differences of histone acetylation levels of H3K9,H3K14,H4K12 and H4K16 in ChIP2 coding region of CyclinD1 and CDK4 genes were not statistically significant between NaF treatment groups (F =0.392,0.823,0.999,0.397,0.705,0.049,1.065,0.196,P > 0.05).Conclusion The histone acetylation of CyclinD1 and CDK4 may not be involved in the transcriptional regulation in human primary osteoblasts treated with fluoride.
Epigenetic progress in obstructive sleep apnea syndrome / 中国综合临床
Tiankuo XUE; Yulan ZHU.
Clinical Medicine of China ; (12): 176-179, 2018.
Obstructive sleep apnea syndrome is a kind of common sleep and respiratory disorder with unknown cause. Epigenetics is a branch of genetics that studies the genetic variation of gene expression under the condition of unchanged nucleotide sequence, including DNA methylation, histone modification, chromatin remodeling,and non coding RNA regulation.
Aberrant histone modification in peripheral blood CD4+ T lymphocytes in oral lichen planus / 口腔疾病防治
SHEN Jun; YIN Cao; WANG Xuan; YE Huiming; JIANG Xiao.
Journal of Prevention and Treatment for Stomatological Diseases ; (12): 365-369, 2018.
Objective@#To investigate the histone acetylation level and histone deacetylase (HDAC) activity of peripheral blood CD4+ T lymphocytes in patients with oral lichen planus (OLP). @*Methods @#Twenty-three OLP patients were selected from August 2016 to January 2017 from the Stomatological Hospital, Southern Medical University. The diagnosis was confirmed by pathology, and the lesions were divided into a nonerosive OLP group (11 cases) and an erosive OLP group (12 cases). Ten healthy sex- and age-matched volunteers served as controls. Immunomagnetic beads were used to separate CD4+ T lymphocytes, and histones and nucleoproteins were extracted. The global histone H3/H4 acetylation levels and HDAC activity of CD4+ T lymphocytes from all subjects were detected by ELISA. The differences between the OLP and control groups were statistically analyzed. @*Results@#Global histone H3 hypoacetylation was observed in the OLP group compared with the control group (u = -2.410, P = 0.012). However, there was no significant difference in the serum CD4+ T lymphocyte histone H4 acetylation level between the OLP and control group (u = -1.412, P = 0.158). HDAC activity was significantly higher in the OLP group than in the healthy control group (F = 5.749, P = 0.023), and much higher HDAC activity was observed in the erosive group than in the nonerosive (P = 0.014) and healthy control groups (P = 0.001). The degree of histone H3 acetylation correlated negatively with increased HDAC activity in the OLP group (rs = -0.771, P < 0.001). There was no correlation between the level of histone H3 acetylation and HDAC activity in the healthy control group (rs = 0.382,P = 0.276). The histone H4 acetylation level in the OLP group showed no correlation with HDAC activity (rs = 0.149, P = 0.498), and the histone H4 acetylation level in the control group also showed no correlation with HDAC activity (rs = 0.527, P = 0.117).@*Conclusion @#Abnormal histone acetylation of CD4+ T lymphocytes in the peripheral blood of patients with OLP was identified and could be related to HDAC activity, suggesting that the epigenetic modification of histone acetylation may play a role in the pathogenesis of OLP.
The effects of sodium butyrate on ethanol-seeking behavior and H3K9 acetylation in NMDA receptor 2B subunit promotor region in the hippocampus of rats / 中华行为医学与脑科学杂志
Yanting ZHANG; Libin ZHANG; Yanqing ZHANG; Yahui XU; Xinsheng GUO; Shilong GUO; Ruiling ZHANG.
Objective To investigate the effects of sodium butyrate on ethanol-seeking behavior and H3K9 acetylation levels in NMDA receptor 2B subunit(NR2B) promoter region in the hippocampus of Wistar rats.To explore the epigenetic mechanism underlying ethanol-seeking behavior.Methods According to random number table,48 male Wistar rats were divided into saline group,sodium butyrate group,ethanol group and sodium butyrate + ethanol group,with 12 rats in each group and administered by intraperitioneal injection respectively.Conditioned place preference (CPP)was used to evaluate the ethanol-seeking behavior.Using Western-blot,real-time PCR and chromatin immunoprecipitation assays,the expression of NR2B protein,NR2BmRNA and the relative level acetylated H3K9 in NR2B promoter region in the hippocampus were determined respectively.Results The CPP test and the CPP score in each group were different (P< 0.05).Compared with the CPP test(261.1 ± 102.2) and the CPP score(48.5±94.6) of saline group,the CPP test ((406.8±109.2),(502.7±72.89)) and the CPP score((198.2± 119.4),(277.5±76.2)) of ethanol group and sodium butyrate + ethanol group were significantly higher(P<0.05),the CPP test(193.4±93.8) and the CPP score (9.7±94.0)of sodium butyrate group were not significantly different(P>0.05).Compared with the ethanol group,CPP test of sodium butyrate + ethanol group was significantly higher(P<0.05).The expression of NR2B protein,NR2BmRNA and acetylated level H3K9 in NR2B promoter region in the hippocampus in each group were different (P< 0.05).Compared with the expression of NR2B protein (1.00 ± 0.28),NR2BmRNA(1.00±0.14) and H3K9 acetylation in NR2B promoter region(1.00±0.25)in the hippocampus of saline group the expression of NR2B protein((1.40±0.34),(1.79±0.30)),NR2BmRNA((1.26±0.16),(1.50±0.08)) and aeetylated level H3K9 in NR2B promoter region ((1.68±0.16),(2.35±0.45)) of ethanol group and sodium butyrate ± ethanol group were significantly higher(P<0.05).The expression of NR2B protein(0.85±0.24),NR2BmRNA(1.05±0.13) and acetylated level H3K9 in NR2B promoter region(0.96±0.41) of sodium butyrate group were not significantly different(P>0.05).Compared with the ethanol group,the expression of NR2B protein,NR2BmRNA and acetylated level H3K9 in NR2B promoter region in the hippocampus of ethanol group,these of sodium butyrate + ethanol group were significantly higher (P<0.05).The CPP score were positively correlated with the expression of NR2B protein (r=0.474,P<0.05).The expression of NR2B protein were positively correlated with the expression of NR2BmRNA (r=0.468,P<0.05).The expression of NR2BmRNA were positively correlated with the expression of H3K9 acetylation in NR2B promoter region(r=0.596,P<0.05),and the CPP score were positively correlated with the expression of H3K9 acetylation in NR2B promoter region (r=0.542,P<0.05).Conclusion The increasing acetylation level of H3K9 in NR2B promoter region in the hippocampus may be one of the epigenetic mechanisms of promoting ethanolseeking behavior,and H3K9 deacetylation in NR2B promoter region in the hippocampus is likely to be a new target for controlling ethanol dependence.
Effects of p300/CBP on histone acetylation of Foxp3 gene in children with Kawasaki disease / 中华微生物学和免疫学杂志
Jiehua MEI; Qin WANG; Guobing WANG; Pengqiang WEN; Mingguo XU; Gen TANG; Dong CUI; Cong LIU; Dongli MA; Chengrong LI.
Chinese Journal of Microbiology and Immunology ; (12): 347-354, 2017.
Objective To investigate the effects of p300/CBP on histone acetylation of Foxp3 gene and its roles in the immunological pathogenesis of Kawasaki disease (KD).Methods Forty-six children with KD and twenty-eight age-matched health children were consented to participate in this study.Co-immunoprecipitation and real-time PCR were performed to detect Foxp3-associated acetylation levels of histone H4 and binding abilities of p300, CBP, pSmad3 (phosphorylated mothers against decapentaplegic homolog 3) and NF-AT (nuclear factor of activated T cells) with Foxp3 gene in CD4+ T cells.The percentages of CD4+CD25high Foxp3+ cells (Treg) and the expression of Foxp3, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), p300, CBP, TGF-βRⅡ (transforming growth factor β receptor Ⅱ) and pLAT1 at protein level were analyzed by flow cytometry.Quantitative real-time PCR was used to measure the expression of Foxp3, IL-10, TGF-β, TGF-βRⅠ, Egr-1 (early growth response protein 1), RARα (retinoic acid receptor α) and PLCγ1 (phospholipase C-γ1) in Treg cells at mRNA level.Plasma concentrations of TGF-β and retinol acid (RA) were measured by enzyme-linked immunosorbent assay.Results (1) The percentages of Treg cells, levels of Foxp3 and molecules associated with suppressive function of Treg cells (TGF-β, IL-10 and CTLA4), acetylation levels of histone H4 associated with promoter, conserved non-coding DNA sequence 1 (CNS1) and CNS2 of Foxp3 gene decreased remarkably during acute KD (P0.05).(2) The levels of p300 and CBP in Treg cells and their binding abilities with Foxp3 gene were down-regulated significantly during acute KD (P0.05).Conclusion Hypoacetylation of histone H4 associated with Foxp3 gene caused by insufficient expression of p300/CBP and their impaired binding abilities might be involved with immune dysfunction in KD.IVIG therapy regulates the expression of p300/CBP and their binding abilities with Foxp3 gene through up-regulating TGF-β signal.
Research on relationship between abnormal regulation of histone acetylation and depression in rats / 中国药理学通报
Haiyan LI; Xiaoyan FU; Ting CUI; Qingsong JIANG; Hongmei QIU.
Chinese Pharmacological Bulletin ; (12): 52-57,58, 2017.
Aim To study the role of histone acetyla-tion and its involvement in the depression-like behav-iors of rats induced by chronic unpredictable stress (CUS ). Methods Thirty male Sprague Dawley (SD ) rats were randomly divided into control group and model group.The method of solitary condition with CUS for consecutive 28 days was used to establish the rat depression model.The open-field test (OFT)and the forced swimming test (FST)were used to evaluate the depressive behaviors of rats;the real time PCR was used to detect the change of HDAC2 mRNA, and Western blot was used to determine the protein expres-sions of H3,H4,acH3 and acH4 in the prefrontal cor-tex and hippocampus of rats.Results Model group showed obvious depression-like behaviors with decrea-sing locomotive activity in OFT (P <0.01 )and in-creasing immobility time in FST (P<0.01),up-regu-lating the mRNA and protein expression of HDAC2 (P<0.0 1 ),and down-regulating the protein expression of acH3 and acH4 (P<0.01)in the prefrontal cortex and hippocampus significantly,compared with control group.Conclusion The mechanism of depressive be-haviors of rats induced by CUS may be associated with down-regulating the level of histone acetylation modifi-cation.
kw:"Histone acetylation"


Another two Republicans, Reps. Peter T. King of New York and Steve Stivers of Ohio voted just for the THUD bill on Thursday.
The Interior-Environment bill would provide $35.9 billion in discretionary spending subject to budget caps for FY 2019 — $300 million more than FY 2018 and $6 billion more than requested.
It also rejects the administration's proposed 43 percent cut to the EPA and keeps funding at the FY 2018 level, while increasing funding for core, non-wildfire functions of the U.S. Forest Service as well as for the National Park Service and Bureau of Land Management.


Perhaps one of the greatest controversies among financial economists is that between classical and behavioral finance. Clearly, economists on both sides have made significant contributions to our understanding of how investors behave and how financial markets work, as is evident in that leading thinkers from both schools have received the Nobel Memorial Prize in Economic Sciences.
So, if both schools have something to contribute, is it possible to reconcile their theories and find a middle ground? I believe that the answer is yes. In fact, I worked with an academic (Yale's Roger Ibbotson) and two of my colleagues at Morningstar (Thomas Idzorek and James Xiong) on a new book published by the CFA Institute Research Foundation called Popularity: A Bridge Between Classical and Behavioral Finance. As the title says, we think that our approach, which we call popularity, provides a bridge between the opposing camps.
The idea of popularity is simple. Investors are willing to pay more for securities with popular characteristics and less for securities with unpopular characteristics. This causes popular stocks to have lower returns and unpopular stocks higher returns. Thus, investors who are willing to hold unpopular stocks will, over the long run, earn higher returns than other investors. Because stocks of smaller companies are less popular than stocks of larger companies, there is a size premium. Similarly, stocks trading at low price/earnings ratios are less popular than stocks trading at high price/earnings ratios, often because they are out of favor, less well-known, or operate in less glamorous industries. Hence, there is a value premium.
The beauty of the idea of popularity is that it applies to all security characteristics, whether concern for them is rational or irrational. Thus, popularity encompasses both rational and behavioral explanations of market phenomena, and serves as a bridge between the rational and behavioral schools of finance.
In "Popularity and Asset Pricing" Idzorek and Ibbotson seek to express the insights of popularity in an equation for the expected return of a security as a linear function of its risk and nonrisk characteristics. What they lacked was a formal theory as to why this would be true. This is where I came in. Making use of my training in microeconomics, I extended the utility function of the capital asset pricing model (CAPM) to include preferences for security characteristics other than risk and expected return. I interpret this extension to be the dimensions of popularity discussed by Idzorek and Ibbotson (2014). I dubbed the resulting model the popularity asset pricing model (PAPM).
In the PAPM, the expected excess return of each security is a linear function of not only its beta, but also of what I call popularity loadings, which measure the popularity of the security based on its characteristics relative to those of the beta-adjusted market portfolio. The popularity loadings are multiplied by market-wide popularity premiums, which are aggregations of the preferences of the investors regarding the nonrisk characteristics. In this way, the market aggregates investor preferences in determining the influence of security characteristics on the expected returns and prices of the securities.
In the CAPM, the market portfolio is optimal for the average investor, so indexing is an optimal strategy. In the PAPM, the market portfolio may not be optimal for any investor, so the case for indexing is diminished.
In the CAPM, each investor holds the market portfolio in combination with the risk-free asset (long or short), based on the investor's risk aversion. In the PAPM, each investor forms a customized portfolio of the risky assets that reflects his or her attitudes toward security characteristics, and combines that with the risk-free asset (long or short). Hence, each investor's portfolio of risky assets is not available in any index fund.
The new book Popularity: A Bridge Between Classical and Behavioral Finance includes everything from the previous work of Roger Ibbotson, Thomas Idzorek, and myself on popularity and the PAPM, and new material including empirical work (thanks to James Xiong). We hope that this work will help tear down walls and build bridges between the classical and behavioral camps.


This par four has one of the most difficult tee shots on the golf course. The tee shot must carry a cattails area and go between the tree lines. Favor the right side as there is no relief to the left. The approach shot to the right avoids the bunker guarding the front left of the green and allows the ball to bounce down to the left from the small knoll on the right side of the green. The two tier green is narrow and 120 feet deep so yardages into the green are important.


We begin tonight in the Garment District at Italian restaurant, Nittis. Blending fashion and food, Nittis offers an energetic environment reimagining the Noir of the 1950s and New York in the 1980s. The family owners behind the restaurant aimed to bring back beloved personal recipes, featuring modern ingredients and stylized presentations. The father, Will Durollari, comes from a family of culinary professionals whose father was also a restaurateur. The sons, Arben and Kuj Durollari, are the masterminds behind the fashion brand BOND with clients including The Weeknd, Anwar Hadid, and Migos. Together they have brought their artistic aesthetic to the concept, design, and kitchen by collaborating on every aspect of the restaurant. Highlights of the menu include Fagottini Porcini, a reimagined Meatball Parmigiana on a rosemary focaccia panini, and a Florentine style NY Strip Florentina.
After your legit fabulous dinner, take the night up a level as you head to the city's highest rooftop lounge, Sky Room, offering 360-degree views of Manhattan. Sip on cocktails like the Sao Tropez with Leblon cachaca, limes, strawberry puree, simple syrup, sour mix, and Juliette rose wine while taking in incredible views of the city. Toast to new heights and the evening ahead as you sit back, relax and enjoy a delicious nightcap.
idk tip: The menu also includes an extensive global rotating wine list from Italy, California and France and cocktails featuring specially infused bourbons and vodkas including Captain Crunch and Fruit Loops.
idk tip: The Sky Room lounge is located on the roof of the Fairfield Inn & Suites by Marriott New York.


Hotel dieu, French Hôtel-dieu, in France, any medieval hospital; the name now refers only to those whose history goes back to the Middle Ages. Many examples from the Gothic period still remain, notably that of Angers (1153–84), the so-called salle des morts at Ourscamp (early 13th century), and that of Tonnerre (c. 1300).
In all of these the most important feature is a vast hall for beds for the sick. In the two earliest the hall is vaulted and divided into three aisles by pillars, on either side of which it was possible to place four rows of beds. At Tonnerre the great hall, nearly 60 feet (18 m) wide and 300 feet (90 m) long, was roofed with wooden trusses and had a wooden barrel-vault ceiling. The beds were in little chambers along the sides, open to supervision from a gallery that ran continuously around the side walls immediately below the windowsills.
At Beaune the hotel dieu, founded in 1443, is of quite different character. It is a two-storied timber structure and occupies three sides of a courtyard. In addition to the halls for the sick, various other rooms for the use of the nuns were furnished.


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Iceland is like no other places we have seen on earth before. The majestic mountain ranges, volcanic geothermal pools, glaciers, hot springs, waterfalls, and many more have all captured our breathe away. We must say Iceland is on the top of our favorite countries. This land of fire and ice offers travelers many things to do and see while exploring this almost untouched land.
Iceland is one of the most remarkable country we visited. It encompasses many scenic attractions that we cannot imagine. We recommended for travelers to do a ring road during the summer months as there are sun light almost all day long. Iceland barely gets dark even at midnight. The roads are in great condition to drive in as long you stay on the main road (Route 1). Even during the drive, you will see many different amazing scenes (pictures below). Iceland is also known for the northern lights which can be seen in the winter months but the day light will be much shorter which limits you from doing much outdoor activities. Be prepared to bring a water-proof jacket as it can rain any time in Iceland. As for accommodations, we stayed at hotels, guest houses, rental cabins, and studio apartments. We planned the trip around the drive around the ring road, therefore the drive will stop at the nearest location with accommodation we can find for that night. Keep in mind that since some accommodations are hundreds of miles from the next one so there are days where the drive is longer than others. Pricing for each room or guesthouse is around $200-350 per night. On average, expect to drive around 5-6 hours a day for 8 day in Iceland. However, the drive was pleasant given the different scenic landscapes to mesmerize in.
The following blog will show you all the sites we visited, including all pictures of each site. You can use this to help you decide which one to visit, as there are countless spots in Iceland and you won't be able to see them all unless you are there for a longer period of time.
Tip: There are a lot of speed cameras across Iceland, drive the posted speed limit. Each ticket is pretty hefty, penalty is around $200+ USD per ticket, even if you only go 10+ kph over the limit. If you do get tickets, call the number on the ticket and ask them nicely to get the fees reduced, it worked for us!
Feel free to ask any questions you may have. Enjoy!
Driving on the ring road all around Iceland is a scenic en-devour in of itself! The sights you'll see on the road are simply breathtaking, take a look at a few photos below to see for yourself!
In May 2015, we organized a group trip with our friends to Iceland. Our plans were to drive around the entire ring road circling the island for 9 days to explore and see as much as we can. We are all astonished of how uniquely beautiful Iceland is and unlike anywhere else in the world. Our flight was on WOW Air departing from Baltimore (BWI) to Reykjavik (KEF). WOW Air is a budget Icelandic airline which has occasionally huge fare sales as low as $99 one-way ticket from the U.S east coast to Reykjavik. Our ticket was $320 per person and they charge extra for checked luggage so we paid an additional $99 for our bag (round-trip).
Actual map of our road trip recorded by Google's My Tracks. Starting in Reykjavik we drove counter-clockwise around the island.
Upon arriving to the airport in the morning, we proceeded to the rental car office. Do keep in mind that cars in Iceland are a lot more compact than cars in the U.S. We rented a car that advertised to fit 7 passengers but there was no way that car could fit all our group and luggage. Cars in Iceland are that small! As a result, we ended up getting 2 separate vehicles because that is the only option that would fit our group + luggage. Car rental is pricey in Iceland ranging from $500-600 for 8 days.
Driving off in the rental cars, we dropped off our luggage at the Studio Apartment in Reykjavik and drive to our first destination – the Blue Lagoon. This is a geothermal spa which is one of the most visited places in Iceland. It is still a must go destination when you're in Iceland. Entrance tickets are about $50 USD and includes a towel and a sample pack of skin products. After enjoying ourselves, we ate at the Lava Restaurant which is located at the site. The food there is not that great and is overpriced, around $50 USD per person. We recommend to get back and eat inside the city if you can.
Studio Apartments in Reykavik – Rooms in Iceland are SUPER small.
Overall, everyone in our group enjoyed a relaxing day at the Blue Lagoon and it was a great start to our trip in Iceland.
Thingvellir National Park – this park is a big lava field known for its historic significance. Inside the park, you will find a diving spot called Silfra which is tectonic boundary between the North American and European plates. In addition, the water in this lake is one of the purest and clearest water in the world, and you can drink straight from it.
Strokkur Geyser – fountain geyser in geothermal area which is often compared to Old Faithful in Yellowstone National Park (USA).
Gullfoss – this is the first waterfall we saw in Iceland which is the most popular one for many tourists visiting Iceland. Keep in mind that as you get closer to most waterfalls, you will get wrenched from the water of the fall so be sure to wear a rain jacket or waterproof jacket.
Seljalandsfoss – this is one of our favorites waterfall in Iceland as you can go right behind the fall to get a different perspective.
Skogafoss – another beautiful waterfall.
On day three, we checked out of our Studio Apartment in Reykjavik and proceeded on to the next destination on the Ring Road.
Dyrhólaey – this is a small peninsula with high cliff and offshore stacks where you can see different breed of birds including Icelandic bird – puffin.
Reynisfjara – this is a black sand pebbles beach with scenic cliffs and caves in unique column formations.
Day 4 – Ice! Lots of Ice!
For us, this is our favorite day of the trip as we saw the most scenic attractions Iceland has to offer. The day starts with a glacier hike excursion of $78/pp in Skaftafell National Park. During this tour, we walked on glaciers, saw ice caves, and captured melted spring water from the glaciers.
Jokulsarlon Glacial Lagoon – this is by far our group favorite spot in Iceland. This is a deep lagoon made up with many broken pieces of glaciers and ice floating right in the lagoon. This is the most picturesque place and definitely a must-see- spot. Our group also took a boat tour right in the lagoon for $35/pp.
Dettifoss – this is known for the most powerful waterfall in Europe. Be prepared to hike for about 1 mile in snow and muddy path to see the fall but it is all worth it.
Krafla and Víti – this is a volcanic crater with blue water. However, it was frozen and covered in snow when we were there. This picture below captured by the drone.
Námafjall – this is a geothermal field with bubbling mud pools, steam vents, hot boiling springs, and distinct sulfur smell.
Our adventure of whale watching in Husavik was definitely an experience to remember. We were all ecstatic getting on the boat despite the warning of rough at sea. Most of us got seasick on the boat even with taking Dramamine but we did see many whales. Whale watching excursion was $65/pp.
Ásbyrgi Canyon – this is a canyon inside a national park lies in the northern part of Iceland.
Goðafoss – another impressive waterfall.
We all went horseback riding along the Black River in North Iceland as a group excursion for $55/pp, then proceeded to our next destination.
Our last full day in Iceland, we explored the Snæfellsnes peninsula where we visited the Kirkjufell mountain and drove around the peninsula coast.
On our last day we spent our time exploring Reykjavik.
Thank you! We really like your blog too, a lot of detailed information!


The left side of a matching pair, "Portiere Cascade I" was originally designed in the early 18th century in Lille. One can tell this by the addition of a French town in the distance, which weren't added to verdures until then. Prior to that, verdures owned their distinctive look to the use of highly detailed foliage. That said, you can still enjoy the amazing attention to detail in this later verdure. The use of colors, combined with the exquisite attention to the littlest detail, is one of the reasons why verdures remain so popular, even to this day. The result is a tapestry with lots of depth and visual interest, tempered by the addition of more traditional landscape elements. All these elements combine to create a tapestry that's truly a fine work of art. This tapestry is lined and has a tunnel for ease of hanging.


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text:
Taxing Issues | STATES IN CRISIS: Problems and Solutions
July 29, 2020 | STATES IN CRISIS: Problems and Solutions
Michael Leachman
Vice President for State Fiscal Policy, CBPP
Michael Leachman is Vice President for State Fiscal Policy at CBPP. He directs the Center's state policy research, overseeing analyses of state policy trends, how federal policy decisions affect states, and state policy choices that improve equity and boost opportunity.
Prior to joining the Center in 2009, Leachman was a policy analyst for nine years at the Oregon Center for Public Policy (OCPP), a member of the State Priorities Partnership. His work at OCPP included research on corporate income taxes, reserve funds, and spending limits.
Earlier in his career, Leachman worked as a community organizer in Chicago and, during graduate school, conducted a range of research projects in collaboration with community organizations.
Leachman holds a Ph.D. in sociology from Loyola University Chicago.
Adam Michel
Senior Policy Analyst, The Heritage Foundation
Adam N. Michel focuses on tax policy and the federal budget as a Senior Policy Analyst in the Grover M. Hermann Center for the Federal Budget at The Heritage Foundation. His research focuses on how taxes impact the well-being and opportunity of Americans.
Michel is published and quoted in outlets such as The New York Times, Wall Street Journal, The Washington Post, Politico, US News and World Report, and The Hill. He has also joined Fox News, CNN, Fox Business, and CNBC to discuss taxes and the economy. Prior to joining Heritage he was a Program Manager for the Spending and Budget Initiative at the Mercatus Center at George Mason University, where he worked on a broad range of topics relating to federal fiscal policy.
Michel received his PhD in economics from George Mason University in Fairfax, VA and a BA in politics from Whitman College in Walla Walla, WA.

redpajama_set_name: RedPajamaCommonCrawl


Sometimes antivirus provides false results over keygens, cracks and patches. That`s all because cracks and keygens sometimes modify your system files. In 99.99% of cases cracks and keygens are safe. However you must take a look AV scan report below and decide for yourself whether to proceed to download or not.
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Dr. Howard F. Vos, 93, of Philadelphia, formerly of New York, died April 8, 2019 at the Springs at Watermark Logan in Philadelphia.
A noted professor of history at the King's College in New York, he received his Ph.D from Northwestern University and his Th.D from Dallas Theological Seminary.
He was a published scholar in the field of history and archaeological studies. Dr Vos was on the faculty of Kings from 1971-1995 and had also served on the faculty at Trinity College and the Moody Bible Institute with concentrations in ancient history and biblical archeology.
In moving to Philadelphia, he became a member of the Tenth Presbyterian Church of Philadelphia and had also been associated with the Museum at the University of Pennsylvania.
Husband of the late Emmagene Vos, who died in 2012; father of the late Valerie Gay who died in 2008, he is survived by his daughter Marcia Zachensky and her husband Peter of Pleasantville NY; four nieces and two nephews.
His graveside service will be Friday 11:00AM at the Arlington Cemetery, Drexel Hill , Pa 19026.
I first met Howard in 1977 as a new student at the King's College. As a history major under Howard and graduating in 1981 it became apparent that he was far more than my professor, but a personal friend and mentor. I went on to raise a family and then train at Moody Aviation as a pilot, later doing missions and humanitarian work in Mexico, and Howard became one of my very first supporters and prayer partners in ministry. He remained a faithful friend and supporter for many years with my work with Brigade Air. Although we only spoke two or three times a year over the next 25 years, every conversation was met with his opening statement, "I still pray for you everyday".
I still use my old New American Standard Bible. On page 2091 of the 1995 edition is an article, "Archaeology and the Bible", written by Howard. Now and then, I would still tease friends that I personally knew one of the authors of my Bible, Howard Vos.
He was an amazing servant of God, scholar, and played an instrumental role in my life and ministry, and will be sorely missed. See you in Glory Howard.
Dr. Vos was a life changing mentor for me. He was excited about history, world events, The King's College, and of course Christ. His passion was infectious, and I caught it full bore. Dr. Vos guided me through graduate school, and gave me advice and counsel during my career in Congress and even in the private sector. He would call me on occassion to inquire about certain legislation, political and world events. He was always learning and teaching - simultaneously. He told me he prayed for me every day, and that he was proud of all I was doing. He always reminded me that the Lord placed me in a position to make a difference.
My father, Bryan Burtch, went to both Wheaton College and Dallas Theological Seminary at the same time as Dr. Vos. (My parents were translators with Wycliffe Bible Translators in Peru and Colombia.) My freshman year, I was so blessed to have my parents in NY near me because they had furlough then, so they were able to attend many of the events. I remember them coming to a parent orientation day and Dad saying "oh, we need to find Dr. Vos as I know him". So we ran into him in the chapel. They both immediately recognized each other even though it was 35+ years later. Everytime Dr. Vos saw me in the 4 years I was at TKC, he would always ask me how my Dad was doing. I took his Western Civilization class and I credit that for a lifelong interest in history, even though I was a music major.


You must be a current member to stand for election as an office bearer. There are membership categories for markets, individuals (ie. stall holders) and industry & supporters.
Memberships are for 12 months. Join online now.
Please advise of you intention to join the AGM web conference, so we know to expect you. Send us an email or call 0413 920 620 to RSVP.


All Lennox homes currently listed for sale in Lennox as of 04/19/2019 are shown below. You can change the search criteria at any time by pressing the 'Change Search' button below.
If you have any questions or you would like more information about any Lennox homes for sale or other Lennox real estate, please contact us and one of our knowledgeable Lennox REALTORS® would be pleased to assist you.
"Lennox Terrace Gardens! Small 17 unit Complex on a Cul-De-Sac. Tri level townhouses. Garage and private Bedroom with en suite bathroom are located on first floor. First level Bedroom is perfect for college student, elderly parents or extra income. Some of these 1st level bedrooms have a private entrance. On the second floor you have a beautiful bright and large living room which adjoins the Large open kitchen, a full size bathroom and a full size laundry room. All Cabinets are custom maple cabinets with granite counter tops. The third floor offers a large private master bedroom with en suite bathroom and two closets. a second bedroom is adjacent with another full bath. Easy low maintenance complex, no yard to clean up! 2 car attached garage, AC/ Central Heat. Stainless steel appliances to be installed ( dishwasher, range and microwave over the range) . Buyer to choose carpet color ( included in price from seller samples ) (hardwood floors from seller samples are optional, buyer to pay difference in price) .Central vacuum system and Alarm system in place. Monthly HOA covers structure Fire insurance, exterior lighting and all maintenance."
"One of The Largest Units in Lennox Terrace Gardens! Small 17 unit Complex on a Cul-De-Sac. Tri level townhouse Style Condo Complex. Garage and private Bedroom with en suite bathroom are located on first floor. First level Bedroom is perfect for college student, elderly parents or extra income. Some of these 1st level bedrooms have a private entrance. On the second floor you have a beautiful bright and large living room which adjoins the Large open kitchen, a full size bathroom and a full size laundry room. All Cabinets are custom maple cabinets with granite counter tops. The third floor offers a large private master bedroom with en suite bathroom and two closets. a second bedroom is adjacent with another full bath. Easy low maintenance complex, no yard to clean up! 2 car attached garage, AC/ Central Heat. Seller will install stainless steel dishwasher, range and microwave over range prior to closing. Buyer may choose carpet color ( included in price from seller samples ) (hardwood floors from seller samples are optional, buyer to pay difference in price). Central vacuum system and Alarm system in place. Monthly HOA covers structure Fire insurance, exterior lighting and all maintenance"
"Lennox Terrace Gardens! Small 17 unit Complex on a Cul-De-Sac. Tri level townhouse style condo development. Garage and private Bedroom with en suite bathroom are located on first floor. First level Bedroom is perfect for college student, elderly parents or extra income. Some of these 1st level bedrooms have a private entrance. On the second floor you have a beautiful bright and large living room which adjoins the Large open kitchen, a full size bathroom and a full size laundry room. All Cabinets are custom maple cabinets with granite counter tops. The third floor offers a large private master bedroom with en suite bathroom and two closets. a second bedroom is adjacent with another full bath. Easy low maintenance complex, no yard to clean up! 2 car attached garage, AC/ Central Heat. Seller will install stainless steel dishwasher, range and microwave over range prior to closing. Buyer may choose carpet color ( included in price from seller samples ) (hardwood floors from seller samples are optional, buyer to pay difference in price). Central vacuum system and Alarm system in place. Monthly HOA covers structure Fire insurance, exterior lighting and all maintenance."
"Located at the end of a cul-de-sac, this new 3-bedroom, 4-bathroom end-unit townhome has a garage and private bedroom with en suite bathroom on the first floor - perfect for a student, in-laws or extra income. On the second floor is a bright and large living room, with 9+ foot high ceilings, wwhich connects to the open kitchen, a full size bathroom and a full size laundry room. The custom-built cabinets are maple and come with granite counter tops. The third floor offers a large private master bedroom with en-suite bathroom and two large closets. A second bedroom is adjacent with another full bathroom. Buyer has option to upgrade flooring and seller will install stainless steel appliances such as the dishwasher, range and microwave over range prior to closing. Monthly HOA covers structure fire insurance, exterior lighting and maintenance. You don't want to miss the opportunity of living in the newly constructed Lennox Terrace Gardens!"


It's nearly summer, and yes, we're screaming for ice cream. The seasonal appearance of the ice cream push cart goes back to the early days of the 20th century. But the timeless allure of a travelling truck of frozen ice cream treats, sandwiches, and shakes remains as strong as ever to young and old alike. Today's ice cream trucks in the USA feature products made with artisanal methods, locavore ingredients and a passion for bringing people together in a common love of sugar, cream and deliciousness.
From ice cream trike, to truck, and now storefront in Avondale, Big Spoon spreads its mission of building community through the common love of small batch, artisan ice cream. Choose from yummy seasonal flavors like Lemon Buttermilk Poppy Seed, Honeysuckle Blackberry, or Cinnamon Toast Crunch.
Enjoy a scoop of the signature black magic in a cone, or go all out with a two-handed sandwich of vanilla ice cream between salted toffee almond grahams. Scoops or pints are served from a roving retro blue 1963 International Harvester van, aptly named Gatsby.
Home of the farm-to-cone, Parfait is committed to housemade everything (including the sprinkles). They use locally sourced ingredients from farms in the Washington area, and grow many of the herbs and berries used in their ice cream, including lavender, mint and rosemary. Enjoy seasonal treats at their best, like Local Strawberry, Peaches & Cream, and vegan apricot sorbet.
Ice cream sur mer! All that sailing in the sun can make one crave cool iced treats. Sea Snacks LLC comes to the maritime rescue with its 20-foot Mako ice cream boat, found in the waters of Casco Bay, Maine. Sea Snacks is stocked with old-school favorites like Klondike bars, Italian ices, and ice cream sandwiches. Boaters should be in search of a colorful pink and blue 'Ice Cream' flag in the Bay.
Charlotte is graced with two soft serve ice cream trucks to meet local demand. The Sticks Truck offers regular ice cream delights, while the Cones Truck has specialty creations, including the Carolina Gold. This sundae features ribbons of chocolate, caramel and white chocolate sauce, topped with crushed homemade vanilla wafers.
Take one beat-up postal van turned food truck and you've got the makings of an ice cream empire. Coolhaus creators paired homemade cookies and ice cream, combining them into 'cool houses'. These innovative sandwiches – 'sammies' – are named after famous architects, like IM Pei-nut Butter, and served in edible wrappers. Coolhaus has 10 mobile ice cream trucks and carts through the U.S., and will scoop for corporate events and weddings too.
Passion for food science is behind the French custard-style ice cream base made from scratch, without chemical stabilizers or fake anything. Local ingredients are seasonally sourced from the Portland Farmers Market. Flavors scooped from a cheerful blue truck can run exotic, from Blackstrap Gingersnap to Thai Rice. The founder's passion began with beer, so naturally, the 21+ only Chocolate Porter Beer Float is a must sip.
Located in Golden Gate Park near museums and the Japanese Tea Garden, Twirl and Dip serves up natural soft serve in a twirly swirl of organic vanilla bean and organic chocolate ice cream. The cones are dipped into dark chocolate, or topped with handmade toppings and sauces. Fresh fruit ice lollies are also available in seasonal flavors like watermelon lemonade.
The bright retro-fitted mail truck serves up all natural and hormone free ice cream on the go. Their signature product is the S'Cream Ball, an ice cream ball on a stick dipped into the chocolate topping of your choice. Also available in a S'Cream Cup, should the stick prove to be a challenging eat.
They do things on a grander scale in NYC! Van Leeuwen has not one but six trucks offering frozen treats with ingredients that are naturally sourced. Unique flavor combinations change seasonally, including Ube with Coconut Jam, Passion Fruit Layer Cake, or Sweet Sticky Black Rice. Interested in trying this at home? The Van Leeuwen Artisan Ice Cream Cookbook shares over 100 vegan and classic recipes and stories of ice cream truck life.
Sing for your sweets in San Jose. Treatbot serves up handmade ice cream with a side of karaoke in a can't-miss-it fire engine red truck. Choices include unique flavors like Eastside Horchata, Macapuno (a mutant Philipino coconut), and the signature 408 – named after a California phone code – that combines caramel ice cream, fudge, and Oreo cookies.
Hello! specializes in Italian-style ice cream (less cream and air than in the regular version), served from a bright 1965 International Metro van named Ingrid. Operating in summer months, Hello! serves up gelato, sorbetto and other Italian desserts in traditional flavors like pistachio, zabaglione, and espresso.


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text:
During August recess, contact Congress members about support for JCPA
David Chavern
From David Chavern, News Media Alliance
A free and diverse press is the backbone of a healthy and vibrant democracy, but local newspapers across the country are under incredible financial pressure and in jeopardy of going out of business. One of the largest problems facing local journalism is that control of access to trustworthy news online has become concentrated on just two platforms: Facebook and Google. There must be a fair exchange for the significant value news publishers' content provides to Google and Facebook so that publishers can continue to invest in high-quality journalism.
In order to help level the playing field, the bipartisan "Journalism Competition and Preservation Act," reintroduced earlier this year in the House and the Senate, would temporarily allow news publishers to collectively negotiate with the tech platforms for fair compensation for the use of their content. Many Members of Congress from both sides of the aisle have already stepped up and expressed strong support for the bill, recognizing its role as a vital solution to the journalism crisis, but more support is needed to get us across the finish line.
ACT NOW: During this August recess, let your Members of Congress know that their constituents support this important legislation.
Journalism is critical to a functioning democracy and the price of doing nothing – the continuing disappearance of local newspapers – is too great. It is more important than ever to take action that supports quality journalism.
In addition to contacting your Member of Congress, please also consider signing our Change.org petition at www.change.org/savelocalnews.
If you have any questions, please contact us at policy@newsmediaalliance.org.


text:
nwaaff
northwest asian american film festival
About NWAAFF
LATE-NIGHT SHORTS PROGRAM: LET'S TALK ABOUT SEX
A series of short films exploring Asian American sexuality, culminating in MASTERS OF THE PILLOW, an unprecedented documentary about UC Davis professor Darrell Hamamoto's attempt to repair the sexual image of Asian American males by producing an Asian American erotic film. Without a doubt, you will have never seen anything quite like it. NOTE: Due to explicit content of a graphic and/or sexual nature, you must be 18 or older to attend this program — no exceptions.
FILMS:
STERILIZED CHERRY by Susan Choi
Seattle premiere Animation, Korea/USA 2002, 3 min.A critique of stereotypical images of Asian women utilizing images of Lolita set against pin-up photos and schoolgirl video clips of the artist.
* Official Selection, Kwangju Biennale 2002 Susan Choi is a multi-media artist based in Los Angeles. Born in Seoul Korea in 1975, she moved to Los Angeles at age 11. Her work investigates the relationships of power negotiated through one's gender, sexuality, and ethnicity.
THE QUEST FOR LENGTH by Gene Rhee
Seattle premiere
Mockumentary, USA 2001, 17 min.A man goes on a quest to increase the size of his penis and finds out more than he bargained for.
* Official Selection, Sundance Film Festival
* Best Comedy Shorts of 2002, American Cinematheque A graduate of the USC film school's Peter Stark Program, Gene Rhee has sold or optioned projects to Dreamworks SKG, Fox Family Channel, and Hallmark Entertainment and is currently attached to direct the feature film FORSAKEN SEOUL for Maverick Films.
ASIAN PRIDE PORN by Greg Pak
Narrative, USA 1999, 3 min.An infomercial spoof in which Tony award-winning playwright David Henry Hwang hawks progressive pornography featuring smart Asian women and sexually empowered Asian men.
* Audience Award, SinCine Film Festival, NYC Greg Pak is an award-winning writer and director. He also edits FilmHelp.com and AsianAmericanFilm.com, and he was recently named one of "25 Filmmakers to Watch" by Filmmaker Magazine.
MASTERS OF THE PILLOW (work-in-progress cut) by James Hou
Documentary, USA 2003, 60 min.Darrell Y. Hamamoto, a professor of Asian American Studies at the University of California, Davis, specializes in media and popular culture. He is also an amateur adult video producer striving to eroticize Asian American sexuality through a yellow porno movement. Interviews of popular Asian American artists are combined with explosive behind-the-scenes footage of the making of Hamamoto's debut film. James Hou was born and raised in San Francisco. His first short fiction film THE PICKUP screened last year at the Hawaii International Film Festival, San Diego Asian Film Festival, Chicago Asian American Showcase and the Hip Hop Film Fest. He is currently developing a feature film which he plans to direct.
SHORTS PROGRAM: DIRECTOR'S CHOICE
CLOSING NIGHT PROGRAM: ROBOT STORIES
Program spotlight: CLOSING NIGHT GALA: SORCERESS OF THE NEW PIANO
Program spotlight: OPENING NIGHT GALA: THE GRACE LEE PROJECT
PANEL DISCUSSION AND SCREENING: SEX, VIOLENCE, AND ROBOTS: THE NEXT WAVE OF ASIAN AMERICAN CINEMA
CALLS FOR ENTRIES
Northwest Asian Weekly
SHORTS PROGRAM: DIRECTOR'S CHOICE


text:
Q: Restart Activity Android I'm working on an Android application that uses AndEngine. I try to restart an activity in this way:
private void restartActivity() {
    startActivity(new Intent(this, TMXTiledMapExample.class));
    finish();
}

This solution works on most devices, but I've realized that not the Nexus4.
In fact, I get the following error :
08-06 11:24:17.416: E/AndroidRuntime(6299): FATAL EXCEPTION: main
08-06 11:24:17.416: E/AndroidRuntime(6299): java.lang.NullPointerException
08-06 11:24:17.416: E/AndroidRuntime(6299):     at org.andengine.ui.activity.BaseGameActivity.onResumeGame(BaseGameActivity.java:222)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at org.andengine.ui.activity.BaseGameActivity$4.run(BaseGameActivity.java:373)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at android.os.Handler.handleCallback(Handler.java:730)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at android.os.Handler.dispatchMessage(Handler.java:92)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at android.os.Looper.loop(Looper.java:137)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at android.app.ActivityThread.main(ActivityThread.java:5103)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at java.lang.reflect.Method.invokeNative(Native Method)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at java.lang.reflect.Method.invoke(Method.java:525)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at com.android.internal.os.ZygoteInit$MethodAndArgsCaller.run(ZygoteInit.java:737)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at com.android.internal.os.ZygoteInit.main(ZygoteInit.java:553)
08-06 11:24:17.416: E/AndroidRuntime(6299):     at dalvik.system.NativeStart.main(Native Method)

A: Try to restart activity like this:
Intent intent = getIntent();
finish();
startActivity(intent);

A: if like this u're close new activity

startActivity(new Intent(this, TMXTiledMapExample.class));
    finish();

just change finish() first

finish();
  startActivity(new Intent(this, TMXTiledMapExample.class));

redpajama_set_name: RedPajamaStackExchange


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text:
apt_packages=""

show_usage() {
    echo "Usage: $1 [-f] [-h]"
    echo "Sets up and updates both python and native dependencies"
    exit 0
}

found_exe() {
    hash "$1" 2>/dev/null
}

found_lib() {
    local found=1
    local paths="$(ld --verbose | grep SEARCH_DIR | tr -s '; ' '\n' | grep -o '/[^"]*')"
    for p in $paths; do
        if [ -e "$p/$1" ]; then
            found=0
        fi
    done
    return $found
}

check_no_root() {
    if [ $(id -u) -eq 0 ]; then
      echo "This script should not be run as root or with sudo."
      exit 1
    fi
}

install_fann() {
    echo "Compiling FANN..."
    if ! found_exe cmake || ! found_exe curl; then
        echo "Please install cmake and curl first."
        exit 1
    fi
    rm -rf /tmp/fann-2.2.0
    curl -L https://github.com/libfann/fann/archive/2.2.0.tar.gz | tar xvz -C /tmp
    local prev_d="$(pwd)"
    cd /tmp/fann-2.2.0
    cmake .
    sudo make install
    cd "$prev_d"
}

create_virtualenv() {
    if [ ! -d ".venv/" ]; then
        python3 -m venv .venv/ --without-pip
        curl https://bootstrap.pypa.io/get-pip.py | .venv/bin/python
    fi
}

install_deps() {
    echo "Installing packages..."

    if found_exe sudo; then
        local SUDO=sudo
    fi

    while read line; do
        local exe=${line%%:*}
        local packages="$(echo ${line##*:} | tr -s ' ')"
        if found_exe $exe; then
            $SUDO $exe $packages
            break
        elif [ "$exe" = "other" ]; then
            if found_exe tput; then
                green="$(tput setaf 2)"
                blue="$(tput setaf 4)"
                reset="$(tput sgr0)"
            fi
            echo
            echo "${green}Could not find package manager"
            echo "${green}Make sure to manually install:${blue} $packages"
            echo $reset
        fi
    done < packages.txt

    if ! found_lib libfann.so; then
        install_fann
    fi
}

install_piwheels() {
    echo "Installing piwheels..."
    echo "
[global]
extra-index-url=https://www.piwheels.org/simple
" | sudo tee -a /etc/pip.conf
}

has_piwheels() { cat /etc/pip.conf 2>/dev/null | grep -qF 'piwheels'; }

hash_dependencies() {
    md5sum requirements.txt packages.txt > .installed
}

file_has_changed() {
    [ "$force_update" ] || [ ! -f .installed ] || ! < .installed grep "$1" | md5sum -c >/dev/null
}

[ "$1" = "-h" ] && show_usage "$0"
[ "$1" = "-f" ] && force_update="true"
check_no_root

set -e

if file_has_changed "packages.txt"; then
    install_deps
fi

create_virtualenv

if file_has_changed "requirements.txt"; then
    .venv/bin/pip install -r requirements.txt
    .venv/bin/pip install -e .
    .venv/bin/pip install -e mycroft_core
fi

arch="$(python -c 'import platform; print(platform.machine())')"
if [ "$arch" = "armv7l" ] && ! has_piwheels; then
    install_piwheels
fi

hash_dependencies

./start.sh setup

redpajama_set_name: RedPajamaGithub


text:
Maps of Morocco
Blank Maps of Morocco
Morocco is formally the Kingdom of Morocco, the northwesternmost nation in the Maghreb region of North Africa. It overlooks the Mediterranean Sea to the north and the Atlantic Ocean to the west, and has land borders with Algeria to the eastern, and also the contested region of Western Sahara to the south. Morocco additionally claims the Spanish exclaves of Ceuta, Melilla and also Peñón de Vélez de la Gomera, and also numerous small Spanish-controlled islands off its shore.
It covers a location of 446,300 km2 (172,300 sq mi) or 710,850 km2 (274,460 sq mi), with a population of approximately 37 million. Its authorities as well as predominant religious beliefs is Islam, as well as the main languages are Arabic and Berber; the Moroccan dialect of Arabic as well as French are additionally widely spoken. Moroccan identification and also culture is a vibrant mix of Berber, Arab, and also European cultures. Its funding is Rabat, while its largest city is Casablanca.
Inhabited given that the Paleolithic Age over 90,000 years ago, the first Moroccan state was developed by Idris I in 788. It was ultimately ruled by a collection of independent empires, reaching its zenith as a local power in the 11th and 12th centuries, under the Almoravid as well as Almohad dynasties, when it regulated most of the Iberian Peninsula and the Maghreb.
In the 15th and also 16th centuries, Morocco encountered outside risks to its sovereignty, with Portugal seizing some territory and also the Ottoman Realm trespassing from the east. The Marinid and also Saadi dynasties otherwise resisted foreign domination, and Morocco was the only North African nation to escape Ottoman rule. The Alaouite dynasty, which rules the nation to today, took power in 1631, and also over the following two centuries broadened polite as well as industrial relationships with the Western globe.
Morocco's critical location near the mouth of the Mediterranean attracted restored European passion; in 1912, France and also Spain separated the nation into particular protectorates, scheduling an international area in Tangier. Following recurring riots and rebellions versus colonial policy, in 1956 Morocco regained its independence and also reunified.
Considering that freedom, Morocco has stayed fairly secure. It has the fifth-largest economy in Africa and also wields significant impact in both Africa as well as the Arab globe; it is considered a center power in global events as well as holds membership in the Arab Organization, the Union for the Mediterranean, as well as the African Union. Morocco is a unitary semi-constitutional monarchy with a chosen parliament.
The executive branch is led by the King of Morocco as well as the prime minister, while legislative power is vested in both chambers of parliament: your house of Reps and also your house of Councillors. Judicial power rests with the Constitutional Court, which might assess the validity of legislations, political elections, and also referenda. The king holds vast exec and also legal powers, specifically over the military, diplomacy and religious events; he can release mandates called dahirs, which have the force of legislation, as well as can likewise liquify the parliament after speaking with the head of state and also the president of the constitutional court.
Below is a blank printable map of Morocco.
This shows a blank practice worksheet of Morocco.
This map shows the basic outline structure of Morocco.
Here you can see a labeled map of Morocco with cities.
Below is a Physical map of Morocco.
Below is a Political map of Morocco.
Below is a Road map of Morocco.
And here, you can see a Locator map of Morocco.


text:
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text:
Lillard has 40, Blazers tie team record for 3s to beat Kings
Damian Lillard scored eight of his season-high 40 points in the final four minutes to rally his Trail Blazers to a 132-126 victory over the Kings in Sacramento.
Published: 11:48 PM PST January 13, 2021
SACRAMENTO, Calif. (AP) - Damian Lillard scored eight of his season-high 40 points in the final four minutes, and the Portland Trail Blazers tied a franchise record with 23 3-pointers to beat the Sacramento Kings 132-126 for their fourth straight victory.
Lillard and CJ McCollum led Portland's shooting barrage from behind the arc with six 3s apiece. Lillard added 13 assists while McCollum had 28 points, 10 assists and seven rebounds. Jusuf Nurkic had a season-best 18 points and 13 rebounds.
The Blazers trailed by 20 midway through the second quarter and were down 105-100 going into the fourth before rallying to win their second consecutive game against the Kings in five days.
De'Aaron Fox had 29 points for Sacramento, who will next host the Los Angeles Clippers on Friday.