NR4A1 (nuclear receptor subfamily 4, group A, member 1)

Review on NR4A1 (nuclear receptor subfamily 4, group A, member 1), with data on DNA, on the protein encoded, and where the gene is implicated

The secreted triose phosphate isomerase of Brugia malayi is required to sustain microfilaria production in vivo

Human lymphatic filariasis is a major tropical disease transmitted through mosquito vectors which take up microfilarial larvae from the blood of infected subjects. Microfilariae are produced by long-lived adult parasites, which also release a suite of excretory-secretory products that have recently been subject to in-depth proteomic analysis. Surprisingly, the most abundant secreted protein of adult Brugia malayi is triose phosphate isomerase (TPI), a glycolytic enzyme usually associated with the cytosol. We now show that while TPI is a prominent target of the antibody response to infection, there is little antibody-mediated inhibition of catalytic activity by polyclonal sera. We generated a panel of twenty-three anti-TPI monoclonal antibodies and found only two were able to block TPI enzymatic activity. Immunisation of jirds with B. malayi TPI, or mice with the homologous protein from the rodent filaria Litomosoides sigmodontis, failed to induce neutralising antibodies or protective immunity. In contrast, passive transfer of neutralising monoclonal antibody to mice prior to implantation with adult B. malayi resulted in 60–70% reductions in microfilarial levels in vivo and both oocyte and microfilarial production by individual adult females. The loss of fecundity was accompanied by reduced IFNγ expression by CD4+ T cells and a higher proportion of macrophages at the site of infection. Thus, enzymatically active TPI plays an important role in the transmission cycle of B. malayi filarial parasites and is identified as a potential target for immunological and pharmacological intervention against filarial infections

Carboxylic Acid- and Amine-Modified Pluronic F127-Based Thermoresponsive Nanogels as Smart Carriers for Brain Drug Delivery

Abegaz Tizazu Andrgie,1 Cheng-Han Liao,2 Tsung-Yun Wu,2,3 Hsueh-Hui Yang,4 Horng-Jyh Harn,5,6 Shinn-Zong Lin,5,7 Yu-Shuan Chen,4,5,8 Hsieh-Chih Tsai2,3,9 1Department of Biotechnology, Debre Berhan University, Debre Berhan, Ethiopia; 2Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei, 106, Taiwan, Republic of China; 3Advanced Membrane Materials Center, National Taiwan University of Science and Technology, Taipei, 106, Taiwan; 4Department of Medical Research, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien, 970, Taiwan; 5Bioinnovation Center, Buddhist Tzu Chi Medical Foundation, Hualien, 970, Taiwan; 6Department of Pathology, Hualien Tzu Chi Hospital, Tzu Chi University, Buddhist Tzu Chi Medical Foundation, Hualien, 970, Taiwan; 7Department of Neurosurgery, Hualien Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Hualien, 970, Taiwan; 8Center for General Education, Tzu Chi University, Hualien, 970, Taiwan; 9R&D Center for Membrane Technology, Chung Yuan Christian University, Taoyuan, 320, TaiwanCorrespondence: Yu-Shuan Chen, Email [email protected] Hsieh-Chih Tsai, Email [email protected]: The blood-brain barrier (BBB) is a critical protective barrier that regulates the exchange of substances between the circulatory system and brain, restricting the access of drugs to brain tissues. Developing novel delivery strategies across the BBB is challenging but crucial. Multifunctional nanogels are promising drug carriers for delivering therapeutic agents to their intended target areas in the brain tissue.Methods: This study introduced carboxylic acid- and amine-modified Pluronic F127 (ADF127 and EDF127)-based thermoresponsive nanogel systems as drug nanocarriers for brain tissues. The release profiles of 3-butylidenephthalide (BP) from the nanogels were investigated in vitro in phosphate-buffered saline (pH 7.4) at 37 °C for 48 h. Additionally, the accumulation of DiR-labeled nanogels in vital organs was observed using fluorescence imaging.Results: A relatively sustained BP release (27%) from ADF127, followed by rapid BP release (39%) from Pluronic F127 within the first 4 h were observed. In vivo studies using the C57BL/6JNarl mouse model showed that intravenously administered BP-loaded copolymeric nanogels exhibited a rapid BP distribution to the liver, spleen, heart, and kidney. DiR fluorescence intensity in the brain increased in the order Pluronic F127 < ADF127 < EDF127 copolymeric nanogels. Although the fluorescence intensity of DiR in the brain tissue was relatively lower than those in other vital organs, the DiR-labeled EDF127 copolymeric nanogels showed approximately 10-fold higher fluorescence intensity.Conclusion: Positively charged drug carrier nanomaterials demonstrate a higher propensity for transfer through the BBB, significantly expanding the applicability of positively charged EDF127 nanogels as nanocarriers for in vivo brain tissue treatment and imaging. Therefore, owing to their increased permeability across the BBB, carboxylic acid- and amine-modified Pluronic F127 nanogels (EDF127 and ADF127) will also offer a promising approach for brain tissue treatment and imaging.Keywords: blood-brain barrier, DiR labeling, permeability, sustained release, nanogel

Effect of a farnesyl transferase inhibitor (R115777) on ductal carcinoma in situ of the breast in a human xenograft model and on breast and ovarian cancer cell growth in vitro and in vivo

INTRODUCTION: The ras pathway is essential for cell growth and proliferation. The effects of R115777, a farnesyl transferase inhibitor, were investigated in cancer cell lines expressing varying levels of growth factor receptors and with differing ras status. Effects on tumour xenografts and human ductal carcinoma in situ (DCIS) of the breast in a xenograft mouse model were also tested. METHOD: In vitro, the concentrations required to reduce cell numbers by 50% (50% inhibitory concentration) were established (MDA-MB231, MCF-7, MCF-7/HER2-18, BT-474, SK-BR3 and SKOV3). Human DCIS was implanted in nude mice or, in separate experiments, cultured cells were injected (MDA-MB231, MCF-7/HER2-18, SKOV3) and allowed to form tumours. Proliferation and apoptosis were determined by immunohistochemistry in xenografts and cell tumours. RESULTS: The 50% inhibitory concentrations varied a hundred-fold, from 39 nmol/l (± 26 nmol/l) for SKBR3 to 5.9 μmol/l(± 0.8 μmol/l) for MDA-MB231. In MCF-7/HER2-18 and SKOV3 cells the levels of tumour growth inhibition were approximately 85% and 40%, respectively. There was a significant decrease in the cell turnover index (CTI; proliferation/apoptosis). In MDA-MB 231 with activated k-ras no inhibition was observed. In treated DCIS xenografts proliferation decreased and apoptosis increased. The CTI ratio between the start and 1 and 2 weeks of treatment were 1.99 and 1.50, respectively, for controls and 0.85 (P = 0.005) and 0.75 (P = 0.08) for treated xenografts. CONCLUSION: Treatment with the farnesyl transferase inhibitor reduced cell growth in vitro and cell tumour growth in vivo. In DCIS treatment resulted in a reduced CTI. R115777 is a promising treatment for breast cancer but the relation between effect and growth factor receptor and ras status has to be established

Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells

Alternative splicing involves differential exon selection of a gene transcript to generate mRNA and protein isoforms with structural and functional diversity. Abnormal alternative splicing has been shown to be associated with malignant phenotypes of cancer cells, such as chemo-resistance and invasive activity. Screening small molecules and drugs for modulating RNA splicing in human hepatocellular carcinoma cell line Huh-7, we discovered that amiloride, distinct from four pH-affecting amiloride analogues, could “normalize” the splicing of BCL-X, HIPK3 and RON/MISTR1 transcripts. Our proteomic analyses of amiloride-treated cells detected hypo-phosphorylation of splicing factor SF2/ASF, and decreased levels of SRp20 and two un-identified SR proteins. We further observed decreased phosphorylation of AKT, ERK1/2 and PP1, and increased phosphorylation of p38 and JNK, suggesting that amiloride treatment down-regulates kinases and up-regulates phosphatases in the signal pathways known to affect splicing factor protein phosphorylation. These amiloride effects of “normalized” oncogenic RNA splicing and splicing factor hypo-phosphorylation were both abrogated by pre-treatment with a PP1 inhibitor. Global exon array of amiloride-treated Huh-7 cells detected splicing pattern changes involving 584 exons in 551 gene transcripts, many of which encode proteins playing key roles in ion transport, cellular matrix formation, cytoskeleton remodeling, and genome maintenance. Cellular functional analyses revealed subsequent invasion and migration defects, cell cycle disruption, cytokinesis impairment, and lethal DNA degradation in amiloride-treated Huh-7 cells. Other human solid tumor and leukemic cells, but not a few normal cells, showed similar amiloride-altered RNA splicing with devitalized consequence. This study thus provides mechanistic underpinnings for exploiting small molecule modulation of RNA splicing for cancer therapeutics