Milli-arcsecond--scale Spectral Properties and Jet Motions in M87

We have combined high resolution VLBI Space Observatory Programme (VSOP) data at 1.6 and 4.8 GHz with Very Long Baseline Array (VLBA) data at higher frequencies and with similar resolutions to study the spectral properties of the core of M87 with milliarcsecond resolution. The VSOP data allow a more accurate measurement of the turn-over frequency, and hence more reliable determination of associated physical parameters of the source. Comparison of the images with previously published images yields no evidence for significant motion of components in the parsec-scale jet. In addition, the brightness temperatures obtained from model-fits to the core are well below the inverse Compton limit, suggesting the radio emission we are observing is not strongly Doppler boosted. Colour version on http://www.vsop.isas.jaxa.jp/survey/publications/m87.ps.gzComment: To appear in PASJ VSOP special issue. Minor correction

EV space suit gloves (passive)

A pair of pressure and thermal insulating overgloves to be used with an Extravehicular (EV) suit assembly was designed, developed, fabricated, and tested. The design features extensive use of Nomex felt materials in lieu of the multiple layer insulation formerly used with the Apollo thermal glove. The glove theoretically satisfies all of the thermal requirements. The presence of the thermal glove does not degrade pressure glove tactility by more than the acceptable 10% value. On the other hand, the thermal glove generally degrades pressure glove mobility by more than the acceptable 10% value, primarily in the area of the fingers. Life cycling tests were completed with minimal problems. The thermal glove/pressure glove ensemble was also tested for comfort; the test subjects found no problems with the thermal glove although they did report difficulties with pressure points on the pressure glove which were independent of the thermal glove

Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum

Conjugated bile salt hydrolase (BSH) catalyses the hydrolysis of the amide bond that conjugates bile acids to glycine and to taurine. The BSH enzyme from Bifidobacterium longum was overexpressed in Escherichia coli BL21(DE3), purified and crystallized. Crystallization conditions were screened using the hanging-drop vapour-diffusion method. Crystal growth, with two distinct morphologies, was optimal in experiments carried out at 303 K. The crystals belong to the hexagonal system, space group P622 with unit-cell parameters a = b = 124.86, c = 219.03 Angstrom, and the trigonal space group P321, with unit-cell parameters a = b = 125.24, c = 117.03 Angstrom. The crystals diffracted X-rays to 2.5 Angstrom spacing. Structure determination using the multiple isomorphous replacement method is in progress

Expression, purification, crystallization and preliminary X-ray diffraction analysis of conjugated bile salt hydrolase from Bifidobacterium longum

Conjugated bile salt hydrolase (BSH) catalyses the hydrolysis of the amide bond that conjugates bile acids to glycine and to taurine. The BSH enzyme from Bifidobacterium longum was overexpressed in Escherichia coli BL21(DE3), purified and crystallized. Crystallization conditions were screened using the hanging-drop vapour-diffusion method. Crystal growth, with two distinct morphologies, was optimal in experiments carried out at 303 K. The crystals belong to the hexagonal system, space group P622 with unit-cell parameters a = b = 124.86, c = 219.03 Angstrom, and the trigonal space group P321, with unit-cell parameters a = b = 125.24, c = 117.03 Angstrom. The crystals diffracted X-rays to 2.5 Angstrom spacing. Structure determination using the multiple isomorphous replacement method is in progress

Comparison of research framing preferences and information use of state legislators and advocates involved in cancer control, United States, 2012–2013

INTRODUCTION: Evidence-based policy plays an important role in prevention of cancer and other chronic diseases. The needs of actors involved in policy decision-making should inform knowledge translation strategies. This study examines the differences between state legislators and advocates in how they seek and use information and what their preferences are for how research information is framed. METHODS: We conducted a cross-sectional comparison of survey responses by US advocates (n = 77) and state legislators (n = 265) working on issues related to cancer control. RESULTS: Advocates differed significantly from legislators on all demographic characteristics. Advocates reported seeking and using information more frequently than legislators, though legislators used legislative research bureaus more often (0.45 point difference, P = .004). Both legislators and advocates prioritized the presentation and timeliness of research information similarly but reported different preferences for source (information bias, information relevance, delivery of information by trusted person) of research information. Several differences between advocates and legislators were modified by participant age. CONCLUSION: Our study provides insights for development of knowledge translation strategies to enhance evidence-based policy making for cancer control that are tailored to state-level legislators and advocates. Additional research efforts should evaluate the effectiveness of such knowledge translation strategies, particularly among advocates

Cloning, preparation and preliminary crystallographic studies of penicillin V acylase autoproteolytic processing mutants

The crystallization of three catalytically inactive mutants of penicillin Vacylase (PVA) from Bacillus sphaericus in precursor and processed forms is reported. The mutant proteins crystallize in different primitive monoclinic space groups that are distinct from the crystal forms for the native enzyme. Directed mutants and clone constructs were designed to study the post-translational autoproteolytic processing of PVA. The catalytically inactive mutants will provide threedimensional structures of precursor PVA forms, plus open a route to the study of enzyme-substrate complexes for this industrially important enzyme

Structure of the T6 Human Nickel Insulin Derivative at 1.35 Å Resolution

This paper presents results of the structural investigation on the human insulin hexamer derivative stabilised by nickel coordination. Single crystals of the Ni-insulin derivative were prepared by the hanging drop vapour diffusion crystallisation method using metal-free insulin and nickel(II) acetate tetrahydrate. The low-temperature crystal structure was determined by the single crystal X-ray crystallographic method with data extending to 1.35 Å spacing. The investigated insulin derivative exhibits the T6 form of insulin and crystallizes in the trigonal system in space group R3, with the unit cell parameters a = b = 81.41 Å and c = 33.75 Å. There are two nickel atoms per insulin hexamer and both are octahedrally coordinated by three Nε2 atoms of three symmetry-related HisB10/HisD10 and three oxygen atoms of three symmetry-related water molecules

How to change the oligomeric state of a circular protein assembly: switch from 11-subunit to 12-subunit TRAP suggests a general mechanism

Many critical cellular functions are performed by multisubunit circular protein oligomers whose internal geometry has evolved to meet functional requirements. The subunit number is arguably the most critical parameter of a circular protein assembly, affecting the internal and external diameters of the assembly and often impacting on the protein's function. Although accurate structural information has been obtained for several circular proteins, a lack of accurate information on alternative oligomeric states has prevented engineering such transitions. In this study we used the bacterial transcription regulator TRAP as a model system to investigate the features that define the oligomeric state of a circular protein and to question how the subunit number could be manipulated.We find that while Bacillus subtilis and Bacillus stearothermophilus TRAP form 11-subunit oligomers, the Bacillus halodurans TRAP exclusively forms 12-subunit assemblies. Significantly, the two states of TRAP are related by a simple rigid body rotation of individual subunits around inter-subunit axes. We tested if such a rotation could be induced by insertion or deletion mutations at the subunit interface. Using wild type 11-subunit TRAP, we demonstrate that removal of five C-terminal residues at the outer side of the inter-subunit axis or extension of an amino acid side chain at the opposite, inner side, increased the subunit number from 11 to 12. Our findings are supported by crystal structures of TRAP oligomers and by native mass spectrometry data.The subunit number of the TRAP oligomer can be manipulated by introducing deletion or addition mutations at the subunit interface. An analysis of available and emerging structural data on alternative oligomeric states indicates that the same principles may also apply to the subunit number of other circular assemblies suggesting that the deletion/addition approach could be used generally to engineer transitions between different oligomeric states

Thermodynamic Contributions of Deoxyuridine Loops to the Folding of DNA Straight Hairpin Loops with d(GCGC/GCGC) Stems

The global stability of DNA molecules depends on base stacking, base-pairing, ionic and hydration contributions. To understand the mechanisms that govern the many biological roles of nucleic acids, it is essential to have a complete physical description of the folding of nucleic acids, including ion and water binding. We used a combination of temperature-dependent UV spectroscopy and differential scanning calorimetry to investigate the effect of the loop length on the unfolding thermodynamics of a set of DNA stem-loop motifs with the following sequence: d(5’-GCGCUnGCGC), where n = 3, 5, 7, and 9. The folding of each hairpin loop resulted in a monophasic transition that was monomolecular. The favorable folding of each hairpin (negative ∆Go) in this set resulted in a thermodynamic compensation of a favorable enthalpy with an unfavorable entropy contributions, enthalpy contributions correspond to formation of base-pair stackings while the unfavorable entropy contributions correspond mainly to the immobilization of counterions, as the length of the uracil hairpin loop increases, the stability of the hairpin decreases (the free energy term becomes more positive), due to a less favorable entropic contribution because more counterions are binding. Relative to a similar set of thymine hairpin loops, we measured less favorable free energy terms because of the lower thermal stability of the uracil hairpins (lower TM’s) and their lesser impact of ion binding. Supported by Grant MCB-1912587 from the NSF.https://digitalcommons.unmc.edu/surp2022/1025/thumbnail.jp