Endogenous retinoids in rat epididymal tissue and rat and human spermatozoa

Recent work has demonstrated high levels of retinoid binding proteins in rat epididymis, and a lumenal retinoic acid binding protein has been purified. These findings suggested that vitamin A may be involved in spermatozoal maturation in the epididymis. We further addressed this question by quantifying retinol, retinyl esters, and retinoic acid isomers from perfused epididymal tissue, from rat testicular and epididymal spermatozoa, and from human ejaculate sperm. HPLC showed vitamin A levels to be higher in caput than in corpus or cauda tissue. Retinoic acid and 9-cis-retinoic acid were found to be graded from lowest levels in caput to highest in cauda. Spermatozoa from caput epididymidis and enriched testicular spermatozoa were found to have higher levels of vitamin A than did spermatozoa from corpus or cauda epididymidis. Spermatozoal retinyl esters had acyl substituents similar to those seen in whole epididymis, and diminished in quantity in sperm from distal segments. Human ejaculate sperm were found to retain high levels of retinyl palmitate and stearate. Retinol and retinoic acid were only marginally detectable in human sperm. Retention of retinoids in mature spermatozoa suggests roles for vitamin A in spermatozoal reproductive physiology beyond the epididymal stage

LAPAS: A SiGe Front End Prototype for the Upgraded ATLAS LAr Calorimeter

We have designed and fabricated a very low noise preamplifier and shaper to replace the existing ATLAS Liquid Argon readout for use at the Large Hadron Collider upgrade (sLHC). IBM’s 8WL 130nm SiGe process was chosen for it’s radiation tolerance, low noise bipolar NPN devices, wide voltage rand and potential use in other sLHC detector subsystems. Although the requirements for the final design can not be set at this time, the prototype was designed to accommodate a 16 bit dynamic range. This was accomplished by using a single stage, low noise, wide dynamic range preamp followed by a dual range shaper. The low noise of the preamp is made possible by the low base spreading resistance of the Silicon Germanium NPN bipolar transistors. The relatively high voltage rating of the NPN transistors is exploited to allow a gain of 650V/A in the preamplifier which eases the input voltage noise requirement on the shaper. Each shaper stage is designed as a cascaded differential operational amplifier doublet with a common mode operating point regulated by an internal feedback loop. Measurement of the fabricated circuits indicates their performance is consistent with the desig

A 5‑lipoxygenase-specific sequence motif impedes enzyme activity and confers dependence on a partner protein

© 2018 Elsevier B.V. Leukotrienes (LT) are lipid mediators of the inflammatory response that play key roles in diseases such as asthma and atherosclerosis. The precursor leukotriene A 4 (LTA 4 ) is synthesized from arachidonic acid (AA) by 5‑lipoxygenase (5-LOX), a membrane-associated enzyme, with the help of 5‑lipoxygenase-activating protein (FLAP), a nuclear transmembrane protein. In lipoxygenases the main chain carboxylate of the C-terminus is a ligand for the non-heme iron and thus part of the catalytic center. We investigated the role of a lysine-rich sequence (KKK 653–655 ) 20 amino acids upstream of the C-terminus unique to 5-LOX that might displace the main-chain carboxylate in the iron coordination sphere. A 5-LOX mutant in which KKK 653–655 is replaced by ENL was transfected into HEK293 cells in the absence and presence of FLAP. This mutant gave ~20-fold higher 5-LOX product levels in stimulated HEK cells relative to the wild-type 5-LOX. Co-expression of the enzymes with FLAP led to an equalization of 5-LOX products detected, with wild-type 5-LOX product levels increased and those from the mutant enzyme decreased. These data suggest that the KKK motif limits 5-LOX activity and that this attenuated activity must be compensated by the presence of FLAP as a partner protein for effective LT biosynthesis

Histidine switch controlling pH-dependent protein folding and DNA binding in a transcription factor at the core of synthetic network devices

© 2016 The Royal Society of Chemistry. Therapeutic strategies have been reported that depend on synthetic network devices in which a urate-sensing transcriptional regulator detects pathological levels of urate and triggers production or release of urate oxidase. The transcription factor involved, HucR, is a member of the multiple antibiotic resistance (MarR) protein family. We show that protonation of stacked histidine residues at the pivot point of long helices that form the scaffold of the dimer interface leads to reversible formation of a molten globule state and significantly attenuated DNA binding at physiological temperatures. We also show that binding of urate to symmetrical sites in each protein lobe is communicated via the dimer interface. This is the first demonstration of regulation of a MarR family transcription factor by pH-dependent interconversion between a molten globule and a compact folded state. Our data further suggest that HucR may be utilized in synthetic devices that depend on detection of pH changes

LAPAS: A SiGe Front End Prototype for the Upgraded ATLAS LAr

We have designed and fabricated a very low noise preamplifier and shaper with a (RC)2 – CR response to replace the existing ATLAS Liquid Argon readout for use at SLHC. IBM’s 8WL 130nm SiGe process was chosen for its radiation tolerance wide voltage range and potential for use in other LHC detector subsystems. The required dynamic range of 15 bits is accomplished by utilization of a single stage, low noise, wide dynamic range preamp connected to a dual range shaper. The low noise of the preamp (~.01nA / √Hz) is achieved by utilizing the process Silicon Germanium bipolar transistors. The relatively high voltage rating of the npn transistors is exploited to allow a gain of 650V/A. With this gain the equivalent input voltage noise requirement on the shaper to about 2.2nV/ √Hz. Each shaper stage is designed as a cascaded differential op amp doublet with a common mode operating point regulated by an internal feedback loop. The shaper outputs are designed to be compatible with the 130nm CMOS ADC being developed in parallel with this effort. Preliminary measurement of the fabricated circuits indicates their performance is consistent with the design specifications

Synthesis of Periodic Mesoporous Silica Thin Films

We have synthesized periodic mesoporous silica thin films from homogeneous solutions. To synthesize the films, a thin layer of a pH 7 micellar coating solution that contains TMOS (tetramethoxysilane) is dip or spin-coated onto Si wafers, borosilicate glass, or quartz substrates. NH3 gas is diffused into the solution and causes rapid hydrolysis and condensation of the TMOS and the formation of periodic mesoporous thin films within 10 seconds. Combination of homogenous solutions and rapid product formation maximizes the concentration of the desired product and provides a controlled, predictable microstructure. The films have been made continuous and crack-free by optimizing initial silica concentration and film thickness. The films are being evaluated as high surface area, size-selective coatings for surface acoustic wave (SAW) sensors

Identification of complex health interventions suitable for evaluation: development and validation of the 8-step scoping framework

Background: There is extensive literature on the methodology of evaluation research and the development and evaluation of complex interventions but little guidance on the formative stages before evaluation and how to work with partner organizations that wish to have their provision evaluated. It is important to be able to identify suitable projects for evaluation from a range of provision and describe the steps required, often with academic institutions working in partnership with external organizations, in order to set up an evaluation. However, research evaluating programs or interventions rarely discusses these stages. Objective: This study aimed to extend work on evaluability assessment and pre-evaluation planning by proposing an 8-Step Scoping Framework to enable the appraisal of multiple programs in order to identify interventions suitable for evaluation. We aimed to add to the literature on evaluability assessment and more recent evaluation guidance by describing the processes involved in working with partner organizations. Methods: This paper documents the steps required to identify multiple complex interventions suitable for process and outcome evaluation. The steps were developed using an iterative approach by working alongside staff in a local government organization, to build an evidence base to demonstrate which interventions improve children’s outcomes. The process of identifying suitable programs for evaluation, thereby establishing the pre-evaluation steps, was tested using all Flying Start provision. Results: The 8-Step Scoping Framework was described using the example of the local government organization Flying Start to illustrate how each step contributes to finding projects suitable for process and outcome evaluation: (1) formulating overarching key questions that encompass all programs offered by an organization, (2) gaining an in-depth understanding of the work and provision of an organization and engaging staff, (3) completing a data template per project/program offered, (4) assessing the robustness/validity of data across all programs, (5) deciding on projects suitable for evaluation and those requiring additional data, (6) negotiating with chosen project leads, both within and outside the organization, (7) developing individual project evaluation protocols, and (8) applying for ethical approval from the university and partner organization. Conclusions: This paper describes the processes involved in identifying suitable projects for evaluation. It adds to the existing literature on the assessment of specific programs suitable for evaluation and guidance for conducting evaluations by establishing the formative steps required to identify suitable programs from a range of provision. This scoping framework particularly relates to academic partners and organizations tasked with delivering evidence-based services designed to meet local needs. The steps identified have been described in the context of early years provision but can be applied to a range of community-based evaluations, or more generally, to cases where an academic partner is working with external stakeholders to identify projects suitable for academic evaluation