Structure–activity relationships of dinucleotides: Potent and selective agonists of P2Y receptors

Dinucleoside polyphosphates act as agonists on purinergic P2Y receptors to mediate a variety of cellular processes. Symmetrical, naturally occurring purine dinucleotides are found in most living cells and their actions are generally known. Unsymmetrical purine dinucleotides and all pyrimidine containing dinucleotides, however, are not as common and therefore their actions are not well understood. To carry out a thorough examination of the activities and specificities of these dinucleotides, a robust method of synthesis was developed to allow manipulation of either nucleoside of the dinucleotide as well as the phosphate chain lengths. Adenosine containing dinucleotides exhibit some level of activity on P2Y1 while uridine containing dinucleotides have some level of agonist response on P2Y2 and P2Y6. The length of the linking phosphate chain determines a different specificity; diphosphates are most accurately mimicked by dinucleoside triphosphates and triphosphates most resemble dinucleoside tetraphosphates. The pharmacological activities and relative metabolic stabilities of these dinucleotides are reported with their potential therapeutic applications being discussed

OFFGAS GENERATION FROM THE DISPOSITION OF SCRAP PLUTONIUM BY VITRIFICATION SIMULANT TESTS

The Department of Energy Office of Environmental Management is supporting R&D for the conceptual design of the Plutonium Disposition Project at the Savannah River Site in Aiken, SC to reduce the attractiveness of plutonium scrap by fabricating a durable plutonium oxide glass form and immobilizing this form within the high-level waste glass prepared in the Defense Waste Processing Facility. A glass formulation was developed that is capable of incorporating large amounts of actinides as well as accommodating many impurities that may be associated with impure Pu feed streams. The basis for the glass formulation was derived from commercial glasses that had high lanthanide loadings. A development effort led to a Lanthanide BoroSilicate (LaBS) glass that accommodated significant quantities of actinides, tolerated impurities associated with the actinide feed streams and could be processed using established melter technologies. A Cylindrical Induction Melter (CIM) was used for vitrification of the Pu LaBS glass. Induction melting for the immobilization of americium and curium (Am/Cm) in a glass matrix was first demonstrated in 1997. The induction melting system was developed to vitrify a non-radioactive Am/Cm simulant combined with a glass frit. Most of the development of the melter itself was completed as part of that work. This same melter system used for Am/Cm was used for the current work. The CIM system used consisted of a 5 inch (12.7 cm) diameter inductively heated platinum-rhodium (Pt-Rh) containment vessel with a control system and offgas characterization. Scrap plutonium can contain numerous impurities including significant amounts of chlorides, fluorides, sodium, potassium, lead, gallium, chromium, and nickel. Smaller amounts of additional elements can also be present. The amount of chlorides present is unusually high for a melter feed. In commercial applications there is no reason to have chloride at such high concentrations. Because the melter operates at 1400-1475 C, many of the impurities present are extremely volatile. An alternative being considered is to pre-treat the impure PuO{sub 2} by water washing to remove the soluble salts, which would significantly reduce the melter emissions. The disadvantage of the washing alternative is the criticality concerns of using water with plutonium. In this paper, the testing that has been conducted at the Savannah River National Laboratory (SRNL) to demonstrate induction melting of impure plutonium simulants will be described. The work described concentrates on quantification of the gaseous and particulate emissions from the induction melter. The Pt-Rh melter vessel is a cylinder with a conical bottom and a tubular drain as shown in Figure 1. A 5-inch (12.7 cm) diameter CIM was used for all of the emissions tests. A 6-inch (15.24 cm) diameter CIM, which is the size of the full-scale melter, has since been constructed for further testing. The 5-inch CIM is heated by three induction coils: one for the 5 inch cylinder, one for the conical section, and one for the 1/4-inch (6.35 mm) drain tube. The 6-inch CIM is similar except the cylinder heater extends lower and also heats the cone. The induction heating system is manufactured by Ameritherm{trademark}. The heating system is controlled by a PC to maintain a specific heat up profile and then maintain a constant energy input that maintains a constant temperature. The CIM is operated in batch mode where the plutonium simulant and the glass-forming frit are first thoroughly mixed in an attrittor mill, then added to the melter. Hafnium oxide (HfO{sub 2}) is used as a simulant for the radioactive PuO{sub 2}. The melter is heated until the mixture begins to melt at about 1100 C, then completely melts at about 1400-1450 C. This temperature is maintained for about three hours. While the temperature is maintained at {approx} 1400 C, an air bubbler is normally used to promote mixing of the glass-forming frit and the waste simulant

Hanford Supplemental Treatment: Literature and Modeling Review of SRS HLW Salt Dissolution and Fractional Crystallization

In order to accelerate waste treatment and disposal of Hanford tank waste by 2028, the Department of Energy (DOE) and CH2M Hill Hanford Group (CHG), Inc. are evaluating alternative technologies which will be used in conjunction with the Waste Treatment Plant (WTP) to safely pretreat and immobilize the tank waste. Several technologies (Bulk Vitrification and Steam Reforming) are currently being evaluated for immobilizing the pretreated waste. Since the WTP does not have sufficient capacity to pretreat all the waste going to supplemental treatment by the 2028 milestone, two technologies (Selective Dissolution and Fractional Crystallization) are being considered for pretreatment of salt waste. The scope of this task was to: (1) evaluate the recent Savannah River Site (SRS) Tank 41 dissolution campaign and other literature to provide a more complete understanding of selective dissolution, (2) provide an update on the progress of salt dissolution and modeling activities at SRS, (3) investigate SRS experience and outside literature sources on industrial equipment and experimental results of previous fractional crystallization processes, and (4) evaluate recent Hanford AP104 boildown experiments and modeling results and recommend enhancements to the Environmental Simulation Program (ESP) to improve its predictive capabilities. This report provides a summary of this work and suggested recommendations

Tenascin-C Enhances Pancreatic Cancer Cell Growth and Motility and Affects Cell Adhesion through Activation of the Integrin Pathway

Background: Pancreatic cancer (PDAC) is characterized by an abundant fibrous tissue rich in Tenascin-C (TNC), a large ECM glycoprotein mainly synthesized by pancreatic stellate cells (PSCs). In human pancreatic tissues, TNC expression increases in the progression from low-grade precursor lesions to invasive cancer. Aim of this study was the functional characterization of the effects of TNC on biologic relevant properties of pancreatic cancer cells. Methods: Proliferation, migration and adhesion assays were performed on pancreatic cancer cell lines treated with TNC or grown on a TNC-rich matrix. Stable transfectants expressing the large TNC splice variant were generated to test the effects of endogenous TNC. TNC-dependent integrin signaling was investigated by immunoblotting, immunofluorescence and pharmacological inhibition. Results: Endogenous TNC promoted pancreatic cancer cell growth and migration. A TNC-rich matrix also enhanced migration as well as the adhesion to the uncoated growth surface of poorly differentiated cell lines. In contrast, adhesion to fibronectin was significantly decreased in the presence of TNC. The effects of TNC on cell adhesion were paralleled by changes in the activation state of paxillin and Akt. Conclusion: TNC affects proliferation, migration and adhesion of poorly differentiated pancreatic cancer cell lines and migh

The coding and non-coding transcriptional landscape of subependymal giant cell astrocytomas

Tuberous sclerosis complex (TSC) is an autosomal dominantly inherited neurocutaneous disorder caused by inactivating mutations in TSC1 or TSC2, key regulators of the mechanistic target of rapamycin complex 1 (mTORC1) pathway. In the CNS, TSC is characterized by cortical tubers, subependymal nodules and subependymal giant cell astrocytomas (SEGAs). SEGAs may lead to impaired circulation of CSF resulting in hydrocephalus and raised intracranial pressure in patients with TSC. Currently, surgical resection and mTORC1 inhibitors are the recommended treatment options for patients with SEGA. In the present study, high-throughput RNA-sequencing (SEGAs n = 19, periventricular control n = 8) was used in combination with computational approaches to unravel the complexity of SEGA development. We identified 9400 mRNAs and 94 microRNAs differentially expressed in SEGAs compared to control tissue. The SEGA transcriptome profile was enriched for the mitogen-activated protein kinase (MAPK) pathway, a major regulator of cell proliferation and survival. Analysis at the protein level confirmed that extracellular signal-regulated kinase (ERK) is activated in SEGAs. Subsequently, the inhibition of ERK independently of mTORC1 blockade decreased efficiently the proliferation of primary patient-derived SEGA cultures. Furthermore, we found that LAMTOR1, LAMTOR2, LAMTOR3, LAMTOR4 and LAMTOR5 were overexpressed at both gene and protein levels in SEGA compared to control tissue. Taken together LAMTOR1-5 can form a complex, known as the 'Ragulator' complex, which is known to activate both mTORC1 and MAPK/ERK pathways. Overall, this study shows that the MAPK/ERK pathway could be used as a target for treatment independent of, or in combination with mTORC1 inhibitors for TSC patients. Moreover, our study provides initial evidence of a possible link between the constitutive activated mTORC1 pathway and a secondary driver pathway of tumour growth

High Diagnostic Performance of Short Magnetic Resonance Imaging Protocols for Prostate Cancer Detection in Biopsy-naive Men: The Next Step in Magnetic Resonance Imaging Accessibility

Background: To make magnetic resonance imaging (MRI) more accessible to men at risk of high-grade prostate cancer (PCa), there is a need for quicker, simpler, and less costly MRI protocols. Objective: To compare the diagnostic performance of monoplanar (“fast” biparametric MRI [bp-MRI]) and triplanar noncontrast bp-MRI with that of the current contrast-enhanced multiparametric MRI (mp-MRI) in the detection of high-grade PCa in biopsy-naïve men. Design, setting, and participants: A prospective, multireader, head-to-head study included 626 biopsy-naïve men, between February 2015 and February 2018. Intervention: Men underwent prebiopsy contrast-enhanced mp-MRI. Prior to biopsy, two blinded expert readers subsequently assessed “fast” bp-MRI, bp-MRI, and mp-MRI. Thereafter, systematic transrectal ultrasound-guided biopsies (SBs) were performed. Men with suspicious mp-MRI (Prostate Imaging Reporting and Data System 3–5 lesions) also underwent MR-in-bore biopsy (MRGB). Outcome measurements and statistical analysis: Primary outcome was the diagnostic performance of each protocol for the detection of high-grade PCa. Secondary outcomes included the difference in biopsy avoidance, detection of low-grade PCa, acquisition times, decision curve analyses, inter-reader agreement, and direct costs. Results from combined MRGB and SB were used as the reference standard. High-grade PCa was defined as grade 2. Results and limitations: Sensitivity for high-grade PCa for all protocols was 95% (180/ 190; 95% confidence interval [CI]: 91–97%). Specificity was 65% (285/436; 95% CI: 61–70%) for “fast” bp-MRI and 69% (299/436; 95% CI: 64–73%) for bp-MRI and mp-MRI. With fast bp-MRI, 0.96% (6/626) more low-grade PCa was detected. Biopsy could be avoided in 47% for the fast bp-MRI and in 49% for the bp-MRI and mp-MRI protocols. Fast bp-MRI and bp-MRI can be performed in 8 and 13 min, respectively, instead of 16