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t the northern Hikurangi margin (North Island, New Zealand), shallow slow slip events (SSEs) frequently accommodate subduction interface plate motion from landward of the trench to

Sialyl-Lewis x (sLex, CD15s) is a tetra-saccharide on the surface of leukocytes required for E-selectin-mediated rolling, a prerequisite for leukocytes to migrate out of the blood vessels. Here we show using flow cytometry that sLex expression on basophils and mast cell progenitors depends on fucosyltransferase 6 (FUT6). Using genetic association data analysis and qPCR, the cell type-specific defect was associated with single nucleotide polymorphisms (SNPs) in the FUT6 gene region (tagged by rs17855739 and rs778798), affecting coding sequence and/or expression level of the mRNA. Heterozygous individuals with one functional FUT6 gene harbor a mixed population of sLex+ and sLex- basophils, a phenomenon caused by random monoallelic expression (RME). Microfluidic assay demonstrated FUT6-deficient basophils rolling on E-selectin is severely impaired. FUT6 null alleles carriers exhibit elevated blood basophil counts and a reduced itch sensitivity against insect bites. FUT6-deficiency thus dampens the basophil-mediated allergic response in the periphery, evident also in lower IgE titers and reduced eosinophil counts., Puan and San Luis et al. find that FUT6, encoding a fucosyltransferase, is required for the “rolling” behavior of certain white blood cells that enables them to move from blood vessels to tissues. They show that FUT6 deficiency leads to a loss of the tetrasaccharide sLex on the surface of basophils, resulting in cells that are less sticky and therefore less able to form the necessary adhesions for exiting the blood vessel to drive the allergic reaction.

To investigate iliotibial band (ITB) diameter thickness at the greater trochanter in patients requiring iliotibial band release who have failed conservative modalities, in comparison to an asymptomatic patient population. A total of 68 subjects were selected to be reviewed using T2 axial plane MRI. The ITB diameter thickness was measured in 34 subjects who underwent surgical ITB release, and compared with a match-paired asymptomatic hip cohort consisting of 34 subjects. ITB diameter thickness was measured at the thickest location for each subject twice by two different examiners. Inter/intra class correlation coefficient was determined for ITB measurement technique accuracy, and the presence of recalcitrant proximal hip pain was evaluated. Interclass correlation coefficient with 95% confidence was measured to be 0.953. The average thickness for ITB surgical release subjects was measured to be 5.61 ± 2.10 mm, and for asymptomatic subjects 3.77 ± 0.79 mm (P

This analysis of Associate Justice Stephen Breyer's jurisprudence proceeds from his first book devoted to this subject, Active Liberty, a term he derives from Benjamin Constant and that Breyer defines as participatory democracy. Active Liberty and two subsequent books, as well as numerous off-bench writings, explain his jurisprudence of pragmatism, an approach he contrasts with originalism. This article addresses three general questions: Is Breyer's jurisprudence, founded on active liberty and pragmatism, fundamentally consistent with the design of the Constitution? Does his jurisprudence support his opinions in the constitutional decisions examined, a number of which are also treated in his books and articles? In a system that is designed to empower and to limit government, do his jurisprudence and judicial decisions constrain judges? This last question is especially important because of Breyer's thesis “that courts should take greater account of the Constitution's democratic nature when they int...

This report documents the acquisition and archiving of a major 3D active-source and passive seismic imaging experiment, NZ3D-FWI. The NZ3D-FWI project aims to image the Hikurangi subduction zone (upper and lower plates and plate boundary fault) along the north Hikurangi margin where shallow slow slip events occur. The primary aim of the project is to collect data optimally to produce high-resolution velocity models using Full-waveform inversion (FWI). The deployment, servicing and collection of the NZ3D-FWI onshore instruments between December 2017 and October 2018 was conducted by Imperial College London, GNS Science, Cardiff University, University of Southampton and Victoria University of Wellington. A 15 x 60 km volume of 3D seismic reflection data was collected offshore of Gisborne by the R/V Marcus Langseth, funded by the National Science Foundation (NSF, USA), the Natural Environment Research Council (NERC, UK) and GNS Science. 99 ocean bottom seismographs were deployed in the 3D seismic reflection volume footprint by JAMSTEC and GNS Science. The onshore NZ3D-FWI deployment included 49 CMG-6TD broadband seismometers from NERC SEIS-UK, which were deployed from December 2017 to October 2018 and detected airgun shots from the 3D seismic reflection survey and local and teleseismic earthquakes over the nine-month period. 119 short-period DATA-CUBE3 instruments loaned from the German Instrument Pool Potsdam (GIPP), and 25 short-period GSX3 instruments from the Earthquake Research Institute (ERI), Tokyo, were deployed between December 2017 and February 2018. These recorded airgun sources and seismicity over a two-month period. The deployment involved a broader array (the Gisborne array), which covered an area of 15 x 30 km with an instrument spacing of ∼2 km. These instruments will primarily be used to construct a high-resolution onshore-offshore velocity model in conjunction with the data collected by the offshore instruments. A denser array of instruments (the Waimata Valley array) with spacing of ∼500 m is aimed at understanding mud volcano processes using active and passive source methods. (auth)

Here, we discuss a relatively new investigative technique useful for identification of human skeletal remains—forensic isotope analysis. Many elements exist in multiple isotope forms, distinguished by differing numbers of neutrons. Stable isotopes contained within human tissues are related to the isotopes in drinking water and food, which can vary based on geography, environment, cultural practices, etc. It's thus possible to analyze the stable isotopes contained within tissues and collect information about someone's life history: where he/she lived, what he/she ate. This chapter presents a brief history of isotope analysis, reviewing foundational research in the fields of ecology, geology, and anthropology that set the stage for the application of stable isotopes in modern forensic investigations of unidentified human remains. It also presents a series of success stories in which forensic isotope analysis helped lead to the identification of an unknown decedent.

The application of plasmid DNA (pDNA)-based gene therapy is limited by its inefficient transgene expression. In this study, minicircle DNA was evaluated for efficient vascular endothelial growth factor (VEGF) expression in skeletal muscle cells. Production of minicircle DNA encoding VEGF was studied by a semi-quantitative electrophoresis method leading to optimized bacterial culture conditions and producing high purity (86.6%) minicircle DNA. The VEGF minicircle DNA under control of the SV40 promoter (pMini-SV-VEGF) showed an increased amount of VEGF mRNA and up to 8 times more VEGF expression than a conventional plasmid (pSV-VEGF) in two different skeletal muscle cell lines (C2C12 and L8). Minicircle DNA with different promoters, including the SV40, CMV and chicken beta-actin, was tested for VEGF expression in a C2C12 skeletal muscle cell line. The high VEGF expression generated by minicircle DNA stimulated efficient endothelial cell growth in vitro. Furthermore, minicircle DNA expressed higher VEGF compared to conventional plasmid in the tibialis anterior (TA) muscle of mice. Taken together, the results suggest that minicircle DNA is an efficacious gene vector for angiogenic VEGF expression in skeletal muscle and may be useful for treating peripheral arterial disease (PAD).

Designing synthetic macromolecular vehicles with high transfection efficiency and low cytotoxicity has been a major interest in the development of non-viral gene carriers. A reducible poly(amido ethylenimine) (SS-PAEI) synthesized by addition copolymerization of triethylenetetramine and cystamine bis-acrylamide (poly(TETA/CBA)) was used as a carrier for small interference RNA (siRNA). Poly(TETA/CBA) could efficiently condense siRNA to form stable complexes under physiological conditions and perform complete release of siRNA in a reductive environment. When formulated with VEGF-directed siRNA, poly(TETA/CBA) demonstrated significantly higher suppression of VEGF than linear-polyethylenimine (PEI) (L-PEI, 25 kDa) in human prostate cancer cells (PC-3). After 5 h of transfection, substantial dissociation and intracellular distribution of siRNA was observed in the poly(TETA/CBA) formulation, but not in the L-PEI formulation. The triggered release of siRNA by reductive degradation of poly(TETA/CBA) in the cytoplasm may affect the RNAi activity by increasing cytoplasmic availability of siRNA. These results suggest that the rational design of non-viral carriers should involve considerations for intracellular dissociation and trafficking of a nucleic acid drug to maximize its effect, in conjunction with formation of stable complexes under physiological conditions.

Apoptosis leads to the fragmentation and packaging of cellular contents into discrete vesicles, a process known as 'blebbing'. Extracellular vesicles express membrane-bound sialic acids, which enable their capture by CD169 (sialoadhesin; Siglec-1) expressing macrophages in the lymph node and spleen. Furthermore, CD169 mediates vesicle trafficking and suppresses the immune response to exosomes-a type of extracellular vesicle released from living cells. In this study, we found that CD169(+) macrophages were the predominant splenic macrophage subset responsible for the capture of EL4 lymphoma-derived apoptotic vesicles (ApoVs) from circulation. CD169(-/-) mice had significantly enhanced in vivo cytotoxic T lymphocyte responses to antigen-pulsed ApoVs, indicating a suppressive role for CD169(+) macrophages to ApoV-associated antigen. In contrast to the observed immunogenic role of ApoVs, the co-administration of unpulsed ApoVs with antigen-pulsed dendritic cells (DCs) significantly suppressed DC-mediated cytotoxic response in vivo; however, this occurred independent of CD169 expression. Overall, our results confirm that apoptosis contributes to both tolerance and immunity, as well as establishing CD169 as a critical mediator of the immune response to extracellular vesicles.

Poly(amido ethylenimine) polymers, a new type of peptidomimetic polymer, containing multiple disulfide bonds (SS-PAEIs) designed to degrade after delivery of plasmid DNA (pDNA) into the cell were synthesized and investigated as new carriers for triggered intracellular gene delivery. More specifically, three SS-PAEIs were synthesized from Michael addition reactions between cystamine bisacrylamide (CBA) and three different ethylene amine monomers, i.e., ethylenediamine (EDA), diethylenetriamine (DETA), or triethylenetetramine (TETA). Complete addition reactions were confirmed by (1)H NMR. The molecular weight, buffer capacity, and relative degree of branching for each SS-PAEI was determined by gel permeation chromatography (GPC), acid-base titration, and liquid chromatography-mass spectroscopy (LC-MS), respectively. Physicochemical characteristics of polymer/pDNA complexes (polyplexes) were analyzed by gel electrophoresis, particle size, and zeta-potential measurements. All three SS-PAEIs effectively complex pDNA to form nanoparticles with diameters less than 200 nm and positive surface charges of approximately 32 mV. The in vitro gene transfer properties of SS-PAEIs were evaluated using mouse embryonic fibroblast cell (NIH3T3), primary bovine aortic endothelial cell (BAEC), and rat aortic smooth muscle cell (A7R5) lines. Interestingly, polyplexes based on all three SS-PAEIs exhibited remarkably high levels of reporter gene expression with nearly 20x higher transfection efficiency than polyethylenimine 25k. The high transfection efficiency was maintained in the presence of 10% serum in the transfection medium. Furthermore, confocal microscopy experiments using labeled pDNA indicated that polyplexes of SS-PAEI displayed greater intracellular distribution of pDNA as compared to PEI, most likely due to environmentally triggered release. Therefore, SS-PAEIs are a new class of transfection agents that facilitate high gene expression while maintaining a low level of toxicity.

Novel ABA triblock copolymers consisting of low molecular weight linear polyethylenimine (PEI) as the A block and poly(ethylene glycol) (PEG) as the B block were prepared and evaluated as polymeric transfectant. The cationic polymerization of 2-methyl-2-oxazoline (MeOZO) using PEG-bis(tosylate) as a macroinitiator followed by acid hydrolysis afforded linear PEI-PEG-PEI triblock copolymers with controlled compositions. Two copolymers, PEI-PEG-PEI 2100-3400-2100 and 4000-3400-4000, were synthesized. Both copolymers were shown to interact with and condense plasmid DNA effectively to give polymer/DNA complexes (polyplexes) of small sizes (100 nm) and moderate zeta-potentials (approximately +10 mV) at polymer/plasmid weight ratiosor =1.5/1. These polyplexes were able to efficiently transfect COS-7 cells and primary bovine endothelial cells (BAECs) in vitro. For example, PEI-PEG-PEI 4000-3400-4000 based polyplexes showed a transfection efficiency comparable to polyplexes of branched PEI 25000. The transfection activity of polyplexes of PEI-PEG-PEI 4000-3400-4000 in BAECs using luciferase as a reporter gene was 3-fold higher than that for linear PEI 25000/DNA formulations. Importantly, the presence of serum in the transfection medium had no inhibitive effect on the transfection activity of the PEI-PEG-PEI polyplexes. These PEI-PEG-PEI triblock copolymers displayed also an improved safety profile in comparison with high molecular weight PEIs, since the cytotoxicity of the polyplex formulations was very low under conditions where high transgene expression was found. Therefore, linear PEI-PEG-PEI triblock copolymers are an attractive novel class of nonviral gene delivery systems.

A new pyrrolopyrimidine alkaloid, rigidin E (1) and the known metabolites rigidin (2) and 1-methylherbipoline (3) have been isolated from a Papua New Guinea tunicate Eudistoma sp. A combination of spectroscopic data were used to determine the structures of these metabolites.

Two cases decided during the 1999-2000 term of the United States Supreme Court addressed a question that has perplexed the Court for more than 50 years: what constitutes an establishment of religion? In Santa Fe v. Doe,1 the Court ruled that student-led prayers at high school football games violated the Establishment Clause. Mitchell v. Helms2 held that a federal program placing computers and other instructional equipment in parochial school classrooms did not violate the clause. The decisions in both of these cases were accompanied by dissenting opinions that presented alternative interpretations of the Establishment Clause.3 In Santa Fe, the majority of the Court reasons that even if the decision to have a pre-game prayer is made by students, the prayer has the effect of coercing those attending the game to participate in an act of religious worship. In his dissent, Chief Justice Rehnquist, joined by Justices Scalia and Thomas, characterizes the Court’s opinion as hostile to religion. Rehnquist recalls that George Washington proclaimed a day of “‘public thanksgiving and prayer, to be observed by acknowledging with grateful hearts the many and signal favors of Almighty God,’” thereby setting a precedent for the support of religion.4 In addition to straying from the original understanding of establishment, the dissenters also believe that because the policy in question has not yet been implemented, the decision is beyond the proper reach of the Court. In addition to Washington’s proclamation of a day of thanksgiving and prayer, other statements from that era support the view that at the time of the founding, religion was regarded as an important aspect of the nation’s political life. In his Farewell Address, Washington stated: “‘Whatever may be conceded to the influence of refined education on minds of peculiar structure, reason and experience both forbid us to expect that national morality can prevail in exclusion of religious principle.’”5 Thomas Jefferson made this statement in his writings: “‘I consider . . . religion a supplement to law in the government of man.’”6 Both of these statements recognize the