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A superconducting transition-edge sensor (TES) microcalorimeter is an ideal X-ray detector for experiments at accelerator facilities because of good energy resolution and high efficiency. To study the performance of the TES detector with a high-intensity pulsed charged-particle beam, we measured X-ray spectra with a pulsed muon beam at the Japan Proton Accelerator Research Complex (J-PARC) in Japan. We found substantial temporal shifts of the X-ray energy correlated with the arrival time of the pulsed muon beam, which was reasonably explained by pulse pileup due to the incidence of energetic particles from the initial pulsed beam.

Aim: The study was aimed at determining the risk assessment of toxic metal concentration in soil and water at two abandoned Lead-Zinc mines Yonov District, Logo, Benue State, Nigeria. Study design: comparative cross-sectional study. Place and duration of study: This study was carried out at the Bruce Powel Toxicology & Biodiversity Laboratory, Animal and Environmental Biology Department, University of Port Harcourt, Analysis at Giolee Global Services Limited Port Harcourt, Nigeria from 16th July to 30th October 2019. Methodology: Eighteen composite water and soil samples were collected and analyzed using Atomic Absorption Spectrophotometer. Results: The mean concentration of Lead (Pb) was higher than that of Zinc (Zn) in all eighteen samples, while, Mercury (Hg) and Cadmium (Cd) were below detection limit in all samples. Mean concentration for Pb in soil samples in the order SiteII>Site I (1.29±0.134 mg/kg>1.26±0.04mg/kg) >control 0.82±0.06 mg/kg. Zn was higher in site I, 0.70±0.10 mg/kg than site II 0.66±0.04mg/kg, and control 0.42±0.02mg/kg. Pb values in water was similar in both sites at 0.46±0.04/0.02 mg/L, while control 0.02±0.001mg/kg. Zn was higher in site II 0.05±0.01mg/L than site I 0.04±0.01Mg/L, control was 0.004±0.00 mg/L. Health risk assessment showed that Estimated daily intake of metals, hazard quotient, hazard index were all less than 1, Carcinogenic risk was within the stipulated ranged of 10-6 to 10-4. Conclusion: Ecological and health risk indices revealed non-contamination by Poisonous metals, however, routine assessment is recommended to forestall any sudden change in the concentration that may result to deleterious effects on human health.

A urea-functionalized chemoreceptor 1,5-bis(2,4-dinitrophenyl)carbonohydrazide (BDC) with versatile applications has been reported in this work. BDC displayed ditopic sensitivity toward toxic indus...

Mevalonate diphosphate decarboxylases (MDDs) catalyze the ATP-dependent-Mg2+-decarboxylation of mevalonate-5-diphosphate (MVAPP) to produce isopentenyl diphosphate (IPP), which is essential in both eukaryotes and prokaryotes for polyisoprenoid synthesis. The substrates, MVAPP and ATP, have been shown to bind sequentially to MDD. Here we report crystals in which the enzyme remains active, allowing the visualization of conformational changes in Enterococcus faecalis MDD that describe sequential steps in an induced fit enzymatic reaction. Initial binding of MVAPP modulates the ATP binding pocket with a large loop movement. Upon ATP binding, a phosphate binding loop bends over the active site to recognize ATP and bring the molecules to their catalytically favored configuration. Positioned substrates then can chelate two Mg2+ ions for the two steps of the reaction. Closure of the active site entrance brings a conserved lysine to trigger dissociative phosphoryl transfer of γ-phosphate from ATP to MVAPP, followed by the production of IPP., Mevalonate diphosphate decarboxylase (MDD) is a key enzyme in the mevalonate pathway and catalyses the decarboxylation of mevalonate-5-diphosphate to isopentenyl diphosphate. Here, the authors provide insights into the conformational changes that occur during substrate binding of MDD and the subsequent enzymatic reaction steps by determining the substrate and intermediate bound crystal structures of Enterococcus faecalis MDD.

A small molecular probe was synthesized and its response towards biologically significant ions such as Zn2+ and F− with low detection threshold (Zn2+: 50 nM and F−: 3 μM) was investigated in the present work. The recognition event was appreciably utilized towards designing a biomolecular logic device. 1H-NMR and ESI-MS analyses suggested a chemodosimetric recognition pathway of the probe in the existence of Zn2+ that was thoroughly explored by theoretical (DFT-D3) analysis. In DFT-D3, to ensure the character of the minimum structures obtained from the geometry optimization, the vibrational frequency was analysed, which further confirmed the absence of imaginary mode of vibration for the substrate, intermediate and product geometries. The presence of only one imaginary mode of vibration for the transition state geometries confirmed the behaviour of the first-order saddle point. The zero-point vibrational energy and thermal correction to the enthalpy and entropy at 298.15 K were also derived at the same level of theory. In case of F−, the pathway follows a supramolecular non-covalent type of interaction. The in vitro recognition of Zn2+ and F− was explored in different cells (Pollens and Candida albicans), which, in turn, establishes the cell permeability and bioapplicability of the present chemosensor. The reversible spectroscopic response of the probe was utilized to design multi logic gate-based circuitry and a biomolecular device with versatile applications that open up new avenues in the realm of supramolecular functional world.

Nanobiotechnology gained at most importance in the current health-care systems. The present study reports the synthesis and characterization of silver and gold nanoparticles which in turn act as a selective inhibitor of human osteosarcoma. The in vitro anticancer studies on U2OS produced IC50 values in the therapeutic range of 29.22 ± 0.42 and 32.83 ± 0.81 μg/mL for silver and gold nanoparticles, while the fibroblast cells (L929) produced IC50 values of 141.26 ± 2.5 and 157.23 ± 2.11 μg/mL for silver and gold nanoparticles, respectively. The growth and action of infectious pathogens can be inhibited by the synthesized nanoparticles. The silver nanoparticles is used to scavenge the hydrogen peroxide and served as a sensor. The time-conserving microwave-assisted nanoparticle fabrication procedure is used here at the natural pH. Silver and gold nanoparticles have surface plasmon resonance peaks at 437 and 553 nm, respectively. The Fourier Transform Infrared spectroscopy fingerprints the role of amide groups of proteins in the synthesis and stabilization process. Transmission electron microscopic analysis proved the spherical geometry of silver and gold nanoparticles with an average size of 20.49 ± 6.68 nm and 25.05 ± 7.31 nm, respectively.

Arsenic induced cleavage of the spirolactam ring of a cleft shaped electronically enriched rhodamine based chemoreceptor molecule 3′,6′-bis(ethylamino)-2-((2-hydroxy-5-methylbenzylidene)amino)-2′,7...

Pancreatic ductal adenocarcinoma (PDAC) is associated with robust activity of the coagulation system. To determine mechanisms by which clotting factors influence PDAC tumor progression, we generated and characterized C57Bl/6-derived KPC (KRasG12D, TRP53R172H) cell lines. Tissue factor (TF) and protease-activated receptor-1 (PAR-1) were highly expressed in primary KPC pancreatic lesions and KPC cell lines similar to expression profiles observed in biopsies of patients with PDAC. In allograft studies, tumor growth and metastatic potential were significantly diminished by depletion of TF or Par-1 in cancer cells or by genetic or pharmacologic reduction of the coagulation zymogen prothrombin in mice. Notably, PAR-1–deleted KPC cells (KPC-Par-1KO) failed to generate sizable tumors, a phenotype completely rescued by restoration of Par-1 expression. Expression profiling of KPC and KPC-Par-1KO cells indicated that thrombin–PAR-1 signaling significantly altered immune regulation pathways. Accordingly, KPC-Par-1KO cells failed to form tumors in immune-competent mice but displayed robust tumor growth comparable to that observed with control KPC cells in immune-compromised NSG mice. Immune cell depletion studies indicated that CD8 T cells, but not CD4 cells or natural killer cells, mediated elimination of KPC-Par-1KO tumor cells in C57Bl/6 mice. These results demonstrate that PDAC is driven by activation of the coagulation system through tumor cell–derived TF, circulating prothrombin, and tumor cell–derived PAR-1 and further indicate that one key mechanism of thrombin/PAR-1–mediated tumor growth is suppression of antitumor immunity in the tumor microenvironment. Significance: The tissue factor–thrombin–PAR-1 signaling axis in tumor cells promotes PDAC growth and disease progression with one key mechanism being suppression of antitumor immunity in the microenvironment.

In Polyethylene terephthalate (PET) processing the raw materials are purified terephthalic acid (PTA), moderately purified terephthalic acid (MTA), monoehalene glycol (MEG). The processing of PTA is very difficult and costly. For reducing PTA percent we are using different percentages of MTA blend in PET processing. MTA affected the properties of polyester and melt spinning process. The properties such as elongation, tenacity, molecular chain length, b-color and IV, thermal properties, % carboxyl groups will change with MTA percentage. MTA percentage also affects fiber breakage percentage s, the melting point of PET. FTIR results show a change in chemical composition. Particle size, 4-CBA content of MTA affects the properties of the fiber.

Nitric oxide (NO) has been implicated as an important neurotransmitter in stress responses and sleep regulatory processes. However, the role of NO in the relationship between stress and sleep remains unclear. The medial septum (MS) and vertical diagonal band (VDB), regions of the basal forebrain involved in sleep regulation, contain nitric oxide synthase (NOS) producing neurons. Additionally, NOS neurons in the dorsal raphe nucleus (DRN) encode information about stress duration. The role of nitrergic neurons in these regions in subserving sex-specific responses to stress and sleep loss has yet to be elucidated. In this study, NADPH-d, an index of NOS activity, was used to examine the effects of acute restraint stress and sleep loss on NOS activity in the MS, VDB, and DRN. We show that NOS activity in response to restraint stress, total sleep deprivation (TSD), and partial sleep restriction (PSR) differs based on sex and region. Initial analysis showed no effect of restraint stress or TSD on NOS activity in the basal forebrain. However, investigation of each sex separately revealed that restraint stress and TSD significantly decrease NOS activity in the MS of females, but not males. Interestingly, the difference in NOS activity between restraint stress and TSD in females was not significant. Furthermore, PSR was not sufficient to affect NOS activity in males or females. These data suggest that restraint stress and sleep loss regulate NOS activation in a sex-dependent manner, and that the NOS stress response in females may be mediated by sleep loss.

Homeotherms maintain a stable internal body temperature despite changing environments. During energy deficiency, some species can cease to defend their body temperature and enter a hypothermic and hypometabolic state known as torpor. Recent advances have revealed the medial preoptic area (MPA) as a key site for the regulation of torpor in mice. The MPA is estrogen-sensitive and estrogens also have potent effects on both temperature and metabolism. Here, we demonstrate that estrogen-sensitive neurons in the MPA can coordinate hypothermia and hypometabolism in mice. Selectively activating estrogen-sensitive MPA neurons was sufficient to drive a coordinated depression of metabolic rate and body temperature similar to torpor, as measured by body temperature, physical activity, indirect calorimetry, heart rate, and brain activity. Inducing torpor with a prolonged fast revealed larger and more variable calcium transients from estrogen-sensitive MPA neurons during bouts of hypothermia. Finally, whereas selective ablation of estrogen-sensitive MPA neurons demonstrated that these neurons are required for the full expression of fasting-induced torpor in both female and male mice, their effects on thermoregulation and torpor bout initiation exhibit differences across sex. Together, these findings suggest a role for estrogen-sensitive MPA neurons in directing the thermoregulatory and metabolic responses to energy deficiency., Torpor is a state of reduced metabolism and body temperature that conserves energy when food is scarce. Here the authors show that estrogen-sensitive neurons in the hypothalamus regulate torpor in mice, maintaining torpor in both sexes but initiating torpor and regulating core temperature differentially across sex.

Twenty-one two-proton knockout (p; 3p) cross sections from neutron-rich nuclei at ~ 250 MeV/nucleon were measured. For the first time, the angular distribution of the three emitted protons were measured in coincidence with the tracker and hydrogen target MINOS, demonstrating that the (p; 3p) kinematics is consistent with two sequential proton-proton collisions within the projectile nucleus. Ratios of (p; 3p) over (p; 2p) inclusive cross sections follow the trend of other many-proton (neutron) removal reactions, further reinforcing the sequential nature of (p; 3p) in neutron-rich nuclei.

Here we present new information on the shape evolution of the very neutron-rich ^{92,94}Se nuclei from an isomer-decay spectroscopy experiment at the Radioactive Isotope Beam Factory at RIKEN. High-resolution germanium detectors were used to identify delayed γ rays emitted following the decay of their isomers. New transitions are reported extending the previously known level schemes. The isomeric levels are interpreted as originating from high-K quasineutron states with an oblate deformation of β∼0.25, with the high-K state in ^{94}Se being metastable and K hindered. Following this, ^{94}Se is the lowest-mass neutron-rich nucleus known to date with such a substantial K hindrance. Furthermore, it is the first observation of an oblate K isomer in a deformed nucleus. This opens up the possibility for a new region of K isomers at low Z and at oblate deformation, involving the same neutron orbitals as the prolate orbitals within the classic Z∼72 deformed hafnium region. From an interpretation of the level scheme guided by theoretical calculations, an oblate deformation is also suggested for the ^{94}Se_{60} ground-state band.

Two chemosensors with varying substitution have been synthesized for selective detection of d10 metal ion analyte Zn2+ and Cd2+ by fluorometric method from aqueous medium at very low limit of detec...

A simple, specific, precise and accurate Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the estimation of Diltiazem Hydrochloride in bulk and pharmaceutical dosage form. The chromatographic determination was performed on isocratic high performance liquid chromatography system of Agilent model no.1220. The separation was conducted by using column of Zorbax [C8 (5µ, 4.6 mm×250)] with mobile phase consisting of buffer and Acetonitrile in the ratio of (60:40). The mobile phase was delivered at the flow rate of 1.0ml/min. The eluent was monitored at wavelength 240 nm and found a sharp and symmetrical peak with retention time of 4.66min. The method was validated for linearity, accuracy, precision, specificity, robustness. Recovery of Diltiazem Hydrochloride was found to be in the range of 98%-102%. The method was found to be linear over the concentration range 50-150 µg/ml with coefficient correlation r2 = 0.995. After developing method, validation parameters were carried out successfully and obtained results were complied with USP monograph.

A dual-channel chromogenic and fluorogenic fused-aromatic-system-based chemoreceptor (2-(benzo[d]thiazol-2-yl)-1-((pyren-8-yl)methylene)hydrazine) (TyM) was designed for ditopic sensitivity towards heavy and transition metal ions (HTMs), such as Cu2+, in an aqueous medium and lethal CN- in a semi-aqueous medium. The chemoreceptor displayed proclivity towards the targeted analytes with a distinct optical response (yellow to colourless in the case of Cu2+ and yellow to bright red for CN-). TyM formed a 2 : 1 adduct with Cu2+ with a detection limit of 40 nM. A 1 : 1 binding stoichiometry was confirmed with the chemoreceptor TyM with CN- in sub-nano molar limit of detection. In addition to sophisticated spectroscopic analysis, such as UV-vis, fluorescence, FTIR, 1H-NMR, 13C-NMR, ESI-MS, and HRMS, the plausible mechanistic course of sensing was also established from a theoretical perspective. The reversible UV-vis response of the chemoreceptor TyM towards CN- and H+ can mimic different molecular logic functions and therefore can be exploited for designing several complex electronic circuits principally based on Boolean Algebra. In vitro fluorescence imaging in male microspores of seed plants (Bohonia Nigalandra) and Monilia Albicans (diploid fungus) with TyM and Cu2+ confirmed the permeability of the chemoreceptor TyM at the cellular level as well as its ability to investigate transition metals, such as Cu2+, in biological samples.

The mevalonate pathway produces isopentenyl diphosphate (IPP), a building block for polyisoprenoid synthesis, and is a crucial pathway for growth of the human bacterial pathogen Enterococcus faecalis. The final enzyme in this pathway, mevalonate diphosphate decarboxylase (MDD), acts on mevalonate diphosphate (MVAPP) to produce IPP while consuming ATP. This essential enzyme has been suggested as a therapeutic target for the treatment of drug-resistant bacterial infections. Here, we report functional and structural studies on the mevalonate diphosphate decarboxylase from E. faecalis (MDDEF). The MDDEF crystal structure in complex with ATP (MDDEF–ATP) revealed that the phosphate-binding loop (amino acids 97–105) is not involved in ATP binding and that the phosphate tail of ATP in this structure is in an outward-facing position pointing away from the active site. This suggested that binding of MDDEF to MVAPP is necessary to guide ATP into a catalytically favorable position. Enzymology experiments show that the MDDEF performs a sequential ordered bi-substrate reaction with MVAPP as the first substrate, consistent with the isothermal titration calorimetry (ITC) experiments. On the basis of ITC results, we propose that this initial prerequisite binding of MVAPP enhances ATP binding. In summary, our findings reveal a substrate-induced substrate-binding event that occurs during the MDDEF-catalyzed reaction. The disengagement of the phosphate-binding loop concomitant with the alternative ATP-binding configuration may provide the structural basis for antimicrobial design against these pathogenic enterococci.

The fatty acid, amino acid and fat soluble vitamin content of magur (Clarias batrachus) and singhi (Heteropneustes fossilis) were assayed from collected samples from different geographical locations. Length and weight of collected samples were (50-235g and 165-300 mm) for magur and (50-210g and 190-255mm) for singhi. The essential amino acid (EAA) and the non-essential amino acid (NEAA) content in magur and singhi did not differ significantly. The Vitamin A and D contents were significantly (P

Herein, the interaction of explosive and pollutant nitroaromatics (epNACs) like 2,4,6-trinitrophenol (TNP) with a small molecular probe has been explored by X-ray crystallography and thermodynamics of the crystallization. The supramolecular host with an unusual crystal packing has been used for the detection of TNP. Fluorescence quenching through the RET–PET–ICT–ACQ mechanism results in a strong pi–pi interaction between the receptor and TNP. Both simulated and experimental FTIR spectroscopies have been performed to investigate the sensing event from solid phase perspective. The enhancement in enthalpy (ΔH) of the host–guest adduct suggests an endothermic reaction suitable for sensing a heat-sensitive explosive like the TNP molecule. On the other hand, entropy (ΔS) enhancement in the crystal formation suggests trapping or releasing of the solvent molecule. In reality, it is noticed from the SCXRD study that acetonitrile undergoes co-crystallization in the host–guest adduct. Ultimate energy (Efinal = Eadduct − Eprecursor) of adduct is lowered up to −919.89 Hartree, which favors the crystallization. The recognition event is highly selective for TNP, Cu2+, and F− among the other analytes, where TNP shows fluorescence turn-off and Cu2+ and F− show turn-on response at different spectroscopic energy levels. Indeed, the receptor proves to be a novel tritopic receptor for selective detection of cation, anion, and neutral analytes with a distinguishable signal. In real time stepping, chromatographic strips have been designed for on-spot chromogenic and fluorogenic detection of the TNP-like explosive molecule for homeland safety.

A urea-based BPC [1,5-bis(perfluorophenyl)carbonohydrazide] molecule with four acidic NH protons has been synthesized by a facile synthetic process. The molecule was found to be a ditopic chemosensor for Cd2+ and F− ions. BPC was synthesized from low-cost starting materials, dinitrophenyl hydrazine and triphosgene. The host–guest interactions between the ions (Cd2+ and F−) were not only confirmed by convenient spectroscopic techniques such as UV-Vis, PL, 1H-NMR, FT-IR, and cyclic voltammetry but also through modern DFT, the results of which were in good agreement with the experimental results. In vitro studies in human cancer cells (HeLa cells) were successfully performed with BPC and Cd2+ using fluorescence microscopy. The reversible UV-Vis response for BPC with F−, OH− and H+ mimics multiple logic functions and can be used for several complex electronic circuits based on logic operations. The pH sensor (BPC) can be further interfaced with suitable circuitry interfaced with appropriate programming for ease of access and enhancement of its practical applications.

Introduction The mechanisms underlying the reciprocal relationship between stress and sleep are unclear. Nitric oxide, a diffusible signaling molecule, plays an important role in physiological stress responses and sleep. The medial septum (MS) and vertical diagonal band (VDB) are sleep regulatory regions of the basal forebrain whose cells express nitric oxide synthase (NOS). In this study, we examined the effects of sleep loss and restraint stress on NOS activation in the MS and VDB. Methods Adult male and female C57BL/J6 mice were randomly assigned to a control, total sleep deprivation (TSD), or restraint stress group. TSD was performed for 6h using gentle handling, and restraint was performed for 6h by immobilizing the mice in a plastic restraint device. Both procedures began at light onset in a 12:12 light:dark cycle. Immediately following the procedures, mice were sacrificed, and NADPH-diaphorase (NADPH-d) was measured in the MS and VDB to determine NOS activity. Results A multivariate ANOVA revealed main effects of TSD and restraint stress on NADPH-d in the MS (F(2,13) = 7.0921, p = 0.011) and VDB (F(2,13)=6.416, p = 0.014) in females. A post hoc analysis showed a significant difference between control and TSD (p = 0.011 (MS), p = 0.014 (VDB)), and between control and restraint (p = 0.032 (MS), p = 0.048 (VDB)), but no significant difference between TSD and restraint. There is a sex difference in NADPH-d in these regions (p < 0.005) that reverses direction following TSD and restraint stress. Conclusion Our findings provide evidence that NOS activity in the basal forebrain may underlie sex differences in stress responses. Since there is no significant difference between the TSD and restraint stress, this suggests that the effect of restraint stress on NOS activation is a result of sleep loss, and not due to induction of a stress mechanism. Support This study was partly funded by R01-NS078410 and UCLA start-up funds.

Exclusive cross sections and momentum distributions have been measured for quasifree one-neutron knockout reactions from a ^{54}Ca beam striking on a liquid hydrogen target at ∼200 MeV/u. A significantly larger cross section to the p_{3/2} state compared to the f_{5/2} state observed in the excitation of ^{53}Ca provides direct evidence for the nature of the N=34 shell closure. This finding corroborates the arising of a new shell closure in neutron-rich calcium isotopes. The distorted-wave impulse approximation reaction formalism with shell model calculations using the effective GXPF1Bs interaction and ab initio calculations concur our experimental findings. Obtained transverse and parallel momentum distributions demonstrate the sensitivity of quasifree one-neutron knockout in inverse kinematics on a thick liquid hydrogen target with the reaction vertex reconstructed to final state spin-parity assignments.

Fifty-five inclusive single nucleon removal cross sections from medium mass neutron-rich nuclei impinging on a hydrogen target at � 250 MeV/nucleon were measured at the RIKEN Radioactive Isotope Beam Factory. Systematically higher cross sections are found for proton removal from nuclei with an even number of protons compared to odd-proton number projectiles for a given neutron separation energy. Neutron removal cross sections display no even-odd splitting contrary to nuclear cascade model predictions. Both effects are understood through simple considerations of neutron separation energies and bound state level densities originating in pairing correlations in the daughter nuclei. These conclusions are supported by comparison with semi-microscopic model predictions,highlighting the enhanced role of low-lying level densities in nucleon removal cross sections from loosely-bound nuclei.

A versatile naphthalene-benzothiazole-based chemoreceptor (BTN) towards selective recognition of Zn2+ and Cd2+ is reported in this work. BTN exhibits chromogenic changes (colorless to pale yellow) in presence of Zn2+ in purely aqueous medium. It recognizes Cd2+ by fluorogenic response (non-fluorescent to blooming green emission). While for bio-hazardous CN−, it undergoes dual-channel chromo-fluorogenic response. ESIPT-PET-CHEF-ICT-C = N isomerization pathway unveils the underlying host-guest sensing mechanism. The reversiblespectroscopic response of BTN towards CN−/H+ has been functionally aligned with the logic gate mimic ensemble. Theoretical studies by DFT and electrostatic potential surface analyses further strengthen the recognition phenomenon. BTN undergoes endogenous CN− detection in natural sources like bitter almonds. FE-SEM analysis of BTN displayed varying morphological responses in the presence of Zn2+ and Cd2+. In vitro Cd2+ and CN− detection in Tecoma Stans and Yeast cells confirms its cell permeability and biocompatibility. Interestingly, RGB-based easy-instant-economic prototype is fabricated that escalates the practical application of BTN towards on-field CN− detection. This work indeed opens up a facile and promising route for fabricating selective and sensitive sensing platforms, with good performance towards multi analytes recognition by varying photophysical signaling.

The recent installation of a Multi-Reflection Time-of-Flight (MR-TOF) isobar separator at the CARIBU facility has the promising potential to significantly improve the mass separation and selection of short-lived neutron-rich beams. Ions cycled in the km-long isochronous trajectories between two electrostatic mirrors can be separated to high levels of mass-resolving power within a short time (tens of ms). The installation process is described and results from the first operation are discussed. Following an optimization of the mirror voltages a mass-resolving power of 6.8 · 10 4 was achieved and a separation of isobars was demonstrated. The higher purity beams provided by the MR-TOF and delivered to the Canadian Penning Trap (CPT) will provide access to further measurements of neutron-rich nuclei along the astrophysical r-process path.

7 pags., 7 figs. -- XXIII International School on Nuclear Physics, Neutron Physics and Applications 22-28 September 2019, Varna, Bulgaria, Within the SEASTAR III campaign at the Radioactive Isotope Beam Factory, at the RIKEN Nishina Center, neutron-rich isotopes in the vicinity of 53K were produced from the fragmentation of the primary 70Zn beam on a 9Be target. After nucleon knockout reactions on the secondary liquid hydrogen MINOS target the known γ rays of the neutron-rich 55Sc isotope were observed (shown in this proceedings) and γ rays from 57,59Sc isotopes have been identified for the first time. The evolution of the occupied nucleon orbitals of these nuclei in the ground and excited state is investigated under the prism of the tensor force., We are grateful to the RIKEN Nishina Center accelerator staff for their work in the primary beam delivery and the BigRIPS team for preparing the secondary beams. This work was supported by the cooperation between TU Darmstadt and the GSI Helmholtz Center for Heavy Ion Research, by the Helmholtz Graduate School for Hadron and Ion Research for FAIR and its abroad program and the BMBF under grant No. 05P19RDFN1. D.S. was supported by projects No. GINOP2.3.3-15-2016-00034 and No. K128947.

Glucocorticoid (GR) and mineralocorticoid receptors (MR) are believed to classically bind DNA as homodimers or MR-GR heterodimers to influence gene regulation in response to pulsatile basal or stress-evoked glucocorticoid secretion. Pulsed corticosterone presentation reveals MR and GR co-occupy DNA only at the peaks of glucocorticoid oscillations, allowing interaction. GR DNA occupancy was pulsatile, while MR DNA occupancy was prolonged through the inter-pulse interval. In mouse mammary 3617 cells MR-GR interacted in the nucleus and at a chromatin-associated DNA binding site. Interactions occurred irrespective of ligand type and receptors formed complexes of higher order than heterodimers. We also detected MR-GR interactions ex-vivo in rat hippocampus. An expanded range of MR-GR interactions predicts structural allostery allowing a variety of transcriptional outcomes and is applicable to the multiple tissue types that co-express both receptors in the same cells whether activated by the same or different hormones., Biotechnology and Biological Sciences Research Council (BBSRC); Neuroendocrinology Charitable Trust; BBSRC Alert 13 Capital Grant; Intramural Research Program of the NIH; National Cancer Institute; Center for Cancer Research (USA); Society for Endocrinology Early Career Grant; Biotechnology and Biological Sciences Research Council International Scientific Interchange Scheme Award; British Society for Neuroendocrinology Project Support Grant

The Saccharomyces cerevisiae heat shock protein Hsp31 is a stress-inducible homodimeric protein that is involved in diauxic shift reprogramming and has glyoxalase activity. We show that substoichiometric concentrations of Hsp31 can abrogate aggregation of a broad array of substrates in vitro. Hsp31 also modulates the aggregation of α-synuclein (αSyn), a target of the chaperone activity of human DJ-1, an Hsp31 homolog. We demonstrate that Hsp31 is able to suppress the in vitro fibrillization or aggregation of αSyn, citrate synthase and insulin. Chaperone activity was also observed in vivo because constitutive overexpression of Hsp31 reduced the incidence of αSyn cytoplasmic foci, and yeast cells were rescued from αSyn-generated proteotoxicity upon Hsp31 overexpression. Moreover, we showed that Hsp31 protein levels are increased by H2O2, in the diauxic phase of normal growth conditions, and in cells under αSyn-mediated proteotoxic stress. We show that Hsp31 chaperone activity and not the methylglyoxalase activity or the autophagy pathway drives the protective effects. We also demonstrate reduced aggregation of the Sup35 prion domain, PrD-Sup35, as visualized by fluorescent protein fusions. In addition, Hsp31 acts on its substrates prior to the formation of large aggregates because Hsp31 does not mutually localize with prion aggregates, and it prevents the formation of detectable in vitro αSyn fibrils. These studies establish that the protective role of Hsp31 against cellular stress is achieved by chaperone activity that intervenes early in the protein misfolding process and is effective on a wide spectrum of substrate proteins, including αSyn and prion proteins.

Proteomic studies rely heavily on the use of liquid chromatography (LC)-mass spectrometry (MS and MS/MS) analyses to provide information about protein composition and function. Profiling the proteome can be the first step to understanding biological pathways, but the challenges scientists face with the complex nature of proteins and proteolysis products can be daunting. Techniques involving fractionation, immunoprecipitation, and phosphopeptide enrichment can simplify complex protein mixtures and enhance the amount of target proteins that are important to the investigator. Emphasis on sample preparation for LC-MS/MS analyses is essential to acquisition of high-quality data for proteomic research. Certain classes of reagents, materials, and contaminants that can be introduced during sample processing may limit the effectiveness of LC-MS/MS analysis. These protocols outline methods for proteolytic digestion of proteins that are compatible with LC-MS/MS, along with procedures that allow for simplification of complex protein matrices.

Loquat leaf extract (LLE) is commonly used in China for a variety of ailments including diabetes. Several recent reports implicate LLE and a sesquiterpene glycoside, one of its components, as being an anti-hyperglycemic agent. However, the underlying mechanism of action of this anti-hyperglycemic agent has not been reported. We have conducted a tracer-based metabolomics study to investigate the effects of sesquiterpene and loquat extract on the balance of flux of central glucose metabolism in HepG2 cells and to compare with those of “insulin sensitizers”, metformin and rosiglitazone. Human hepatoma HepG2 cells in confluence culture were incubated in Dulbecco’s modified Eagle’s medium containing 50% [1, 2 13C2]-glucose in the presence of rosiglitazone, metformin, LLE or pure sesquiterpene. Cells were harvested in 48 h. Mass isotopomers of metabolites (glycogen, ribose, deoxyribose, glutamate and palmitate) were determined. 13C labeling in metabolic intermediates were summarized in a mass isotopomer matrix. Treatment with loquat extract/sesquiterpene, metformin and rosiglitazone each produced distinctive mass isotopomer patterns reflecting disparate effects on the contribution of glucose to various metabolites production, and on several metabolic flux ratios. The overall effect of LLE and sesquiterpene on glucose metabolism is clearly different from those of the known “insulin sensitizers”. Our study demonstrates the utility of isotopomer matrix in summarizing metabolic actions of LLE on the balance of fluxes occurring within the central glucose metabolism in HepG2 cells. 13C carbon tracing (tracer-based metabolomics) is a useful systems biology tool to elucidate glucose metabolic pathways affected by diabetes and its treatment.

The catalytic domains of aromatic amino acid hydroxylases (AAAHs) contain a non-heme iron coordinated to a 2-His-1-carboxylate facial triad and two water molecules. Asp139 from Chromobacterium violaceum PAH (cPAH) resides within the second coordination sphere and contributes key hydrogen bonds with three active site waters that mediate its interaction with an oxidized form of the cofactor, 7,8-dihydro-l-biopterin, in crystal structures. To determine the catalytic role of this residue, various point mutants were prepared and characterized. Our isothermal titration calorimetry (ITC) analysis of iron binding implies that polarity at position 139 is not the sole criterion for metal affinity, as binding studies with D139E suggest that the size of the amino acid side chain also appears to be important. High-resolution crystal structures of the mutants reveal that Asp139 may not be essential for holding the bridging water molecules together, because many of these waters are retained even in the Ala mutant. However, interactions via the bridging waters contribute to cofactor binding at the active site, interactions for which charge of the residue is important, as the D139N mutant shows a 5-fold decrease in its affinity for pterin as revealed by ITC (compared to a 16-fold loss of affinity in the case of the Ala mutant). The Asn and Ala mutants show a much more pronounced defect in their kcat values, with nearly 16- and 100-fold changes relative to that of the wild type, respectively, indicating a substantial role of this residue in stabilization of the transition state by aligning the cofactor in a productive orientation, most likely through direct binding with the cofactor, supported by data from molecular dynamics simulations of the complexes. Our results indicate that the intervening water structure between the cofactor and the acidic residue masks direct interaction between the two, possibly to prevent uncoupled hydroxylation of the cofactor before the arrival of phenylalanine. It thus appears that the second-coordination sphere Asp residue in cPAH, and, by extrapolation, the equivalent residue in other AAAHs, plays a role in fine-tuning pterin affinity in the ground state via deformable interactions with bridging waters and assumes a more significant role in the transition state by aligning the cofactor through direct hydrogen bonding.

Cellulose microfibrils are para-crystalline arrays of several dozen linear (1→4)-β-d-glucan chains synthesized at the surface of the cell membrane by large, multimeric complexes of synthase proteins. Recombinant catalytic domains of rice (Oryza sativa) CesA8 cellulose synthase form dimers reversibly as the fundamental scaffold units of architecture in the synthase complex. Specificity of binding to UDP and UDP-Glc indicates a properly folded protein, and binding kinetics indicate that each monomer independently synthesizes single glucan chains of cellulose, i.e., two chains per dimer pair. In contrast to structure modeling predictions, solution x-ray scattering studies demonstrate that the monomer is a two-domain, elongated structure, with the smaller domain coupling two monomers into a dimer. The catalytic core of the monomer is accommodated only near its center, with the plant-specific sequences occupying the small domain and an extension distal to the catalytic domain. This configuration is in stark contrast to the domain organization obtained in predicted structures of plant CesA. The arrangement of the catalytic domain within the CesA monomer and dimer provides a foundation for constructing structural models of the synthase complex and defining the relationship between the rosette structure and the cellulose microfibrils they synthesize.

Five polyrotaxanes were synthesized by threading 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) onto a variety of α,ω-ditriethylenediamino-N-carbamoyl-poly-(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethylene oxide) (Pluronic) triblock copolymers using a two-pot strategy under heterogeneous, nonaqueous conditions. The threaded HP-β-CD units were retained on the pseudopolyrotaxane precursors by end-capping the branched diamine termini with sodium 2,4,6-trinitrobenzene sulfonate. Inclusion of the Pluronic copolymers within the HP-β-CD cavities was more favorable in nonpolar solvents, such as diethyl ether and n-hexane, both of which gave better coverage ratios than polar solvents. (1)H NMR and MALDI-TOF were used to estimate the average molecular weights of the purified polyrotaxane products. A globular morphology of aggregated polyrotaxanes was observed by tapping-mode AFM imaging of dried samples. Treatment of Niemann-Pick C (NPC) type 2-deficient fibroblasts with the polyrotaxane derivatives produced substantial reductions in sterol accumulation, as seen by diminished filipin staining in these cells, suggesting that Pluronic-based polyrotaxanes may be promising vehicles for delivery of HP-β-CD to cells with abnormal cholesterol accumulation.

Phenylalanine hydroxylase (PAH) is a non-heme iron enzyme that catalyzes oxidation of phenylalanine to tyrosine, a reaction that must be kept under tight regulatory control. Mammalian PAH has a regulatory domain in which binding of the substrate leads to allosteric activation of the enzyme. However, the existence of PAH regulation in evolutionarily distant organisms, for example some bacteria in which it occurs, has so far been underappreciated. In an attempt to crystallographically characterize substrate binding by PAH from Chromobacterium violaceum, a single-domain monomeric enzyme, electron density for phenylalanine was observed at a distal site 15.7 Å from the active site. Isothermal titration calorimetry (ITC) experiments revealed a dissociation constant of 24 ± 1.1 μM for phenylalanine. Under the same conditions, ITC revealed no detectable binding for alanine, tyrosine, or isoleucine, indicating the distal site may be selective for phenylalanine. Point mutations of amino acid residues in the distal site that contact phenylalanine (F258A, Y155A, T254A) led to impaired binding, consistent with the presence of distal site binding in solution. Although kinetic analysis revealed that the distal site mutants suffer discernible loss of their catalytic activity, X-ray crystallographic analysis of Y155A and F258A, the two mutants with the most noticeable decrease in activity, revealed no discernible change in the structure of their active sites, suggesting that the effect of distal binding may result from protein dynamics in solution.

Metallothionein (MT) is a cysteine-rich, low molecular weight metal sequestering protein that has been shown to be involved in essential metal homeostasis and in detoxification of heavy metals. In the study sequences of MT genomic DNA and cDNA of freshwater murrel, Channa punctata (Bloch) were reported. cDNA was verified as the C. punctata MT gene by the characteristics of its predicted translation product, namely a high cysteine content, conserved Cys-Cys, Cys-X-Y-Cys, and Cys-X-Cys motifs, and a molecular weight of around 6 kDa. The quantitative real-time reverse-transcriptase polymerase chain reaction (q-RT-PCR) study was carried out in liver, kidney and gill tissues after exposure of the organism to waterborne cadmium for 8 h. At all the time points (viz. 1, 2, 4 & 8 h), a significant up regulation of MT transcript level was observed only in the liver. The results of time kinetics study indicated that expression pattern of C. punctata MT mRNA was found to be in tissue specific manner and the transcript level in the liver could be used as a biomarker of heavy metal contamination in water bodies.

The cation-permeable channel PKD2L1 forms a homomeric assembly as well as heteromeric associations with both PKD1 and PKD1L3, with the cytoplasmic regulatory domain (CRD) of PKD2L1 often playing a role in assembly and/or function. Our previous work indicated that the isolated PKD2L1 CRD assembles as a trimer in a manner dependent on the presence of a proposed oligomerization domain. Herein we describe the 2.7Å crystal structure of a segment containing the PKD2L1 oligomerization domain which indicates that trimerization is driven by the β-branched residues at the first and fourth positions of a heptad repeat (commonly referred to as "a" and "d") and by a conserved R-h-x-x-h-E salt bridge motif that is largely unique to parallel trimeric coiled coils. Further analysis of the PKD2L1 CRD indicates that trimeric association is sufficiently strong that no other species are present in solution in an analytical ultracentrifugation experiment at the lowest measurable concentration of 750nM. Conversely, mutation of the "a" and "d" residues leads to formation of an exclusively monomeric species, independent of concentration. Although both monomeric and WT CRDs are stable in solution and bind calcium with 0.9μM affinity, circular dichroism studies reveal that the monomer loses 25% more α-helical content than WT when stripped of this ligand, suggesting that the CRD structure is stabilized by trimerization in the ligand-free state. This stability could play a role in the function of the full-length complex, indicating that trimerization may be important for both homo- and possibly heteromeric assemblies of PKD2L1.

Buckingham π theorem was applied to model polypyrrole synthesis done by inverse emulsion polymerization route using Methanesulfonic acid as a dopant and potassium persulfate as an oxidant. A list of independent parameters on which polymerization depends was selected and by means of Buckingham theorem a functional relation between three dimensionless quantities; percent yield, conductivity and temperature was obtained. The actual form of the function was determined on the basis of experimental analysis and linear regression analysis. The accuracy of the proposed application was scrutinized by synthesizing pristine polypyrrole salt by the said polymerization route. It was corroborated from the experiment that the increase in percent yield and conductivity was directly proportional to the increase in oxidant concentration. Conductivity as high as 3.77 S/cm was obtained with a maximum percent yield of 94.54% at a moderate temperature range (293 K-298 K).

Polypyrrole, an Intrinsically Conductive Polymer (ICP) was obtained through chemical oxidative polymerization of pyrrole and a low temperature sensing device was designed by pelletizing Ppy/Piezoceramic powder composite with Tin. Raw/Green pellets of Ppy/PZT composite were liquid phase sintered above the solidus temperature of Tin metal (2540C) to yield the final sensor. Current-Voltage (I-V) characteristics of two such sensor samples were analyzed. Sensor sample with 4 % Tin yielded maximum conductivity of 9.12 S/cm as compared with the sensor sample with 3 % Tin, conductivity dropped to 5 S/cm. Also, almost non- linear trend in the I-V curves was observed due to the environmental factors such as a small temperature range between 00C-1100C. Characterization of both the sensor samples using SEM confirmed that the sensor with higher Tin content behaved as a better sensor than the sensor having a lower Tin content.

AMSH plays a critical role in the ESCRT (endosomal sorting complexes required for transport) machinery, which facilitates the down-regulation and degradation of cell-surface receptors. It displays a high level of specificity toward cleavage of Lys63-linked polyubiquitin chains, the structural basis of which has been understood recently through the crystal structure of a highly related, but ESCRT-independent, protein AMSH-LP (AMSH-like protein). We have determined the X-ray structure of two constructs representing the catalytic domain of AMSH: AMSH244, the JAMM (JAB1/MPN/MOV34)-domain-containing polypeptide segment from residues 244 to 424, and AMSH219(E280A), an active-site mutant, Glu280 to Ala, of the segment from 219 to 424. In addition to confirming the expected zinc coordination in the protein, the structures reveal that the catalytic domains of AMSH and AMSH-LP are nearly identical; however, guanidine-hydrochloride-induced unfolding studies show that the catalytic domain of AMSH is thermodynamically less stable than that of AMSH-LP, indicating that the former is perhaps structurally more plastic. Much to our surprise, in the AMSH219(E280A) structure, the catalytic zinc was still held in place, by the compensatory effect of an aspartate from a nearby loop moving into a position where it could coordinate with the zinc, once again suggesting the plasticity of AMSH. Additionally, a model of AMSH244 bound to Lys63-linked diubiquitin reveals a type of interface for the distal ubiquitin significantly different from that seen in AMSH-LP. Altogether, we believe that our data provide important insight into the structural difference between the two proteins that may translate into the difference in their biological function.

An improved synthetic route to polypyrrole salt was accomplished using methanesulfonic acid as a novel dopant along with potassium persulfate as the oxidant employing inverted emulsion polymerization technique. Polypyrrole salt was obtained in high-weight percent yield (83.77%) with respect to the amount of monomer used and the reaction time was drastically reduced (1 h 10 min.) as compared to the previously reported synthesis methods. Characterization of the salt yielded satisfactory results. Inverted emulsion polymerization technique has several unique and distinct advantages over conventional techniques which facilitate the synthesis of fused five membered heterocyclic rings which are otherwise difficult to synthesize, and hence sparsely reported in the literature.

Phenylalanine hydroxylase (PAH), a non-heme iron enzyme, is responsible for the phenylalanine conversion to tyrosine. Its malfunction causes phenylketonuria (PKU). To better understand how protein structure and folding profiles are affected by the metal cofactor, we investigated the chemical (un)folding of apo- and holo-PAH from Chromobacterium violaceum (cPAH) using circular dichroism (CD) and analytical ultracentrifugation (AUC). Holo-cPAH shows a two-state unfolding transition. In contrast, the unfolding profile for apo-cPAH reveals a three-state (un)folding pathway and accumulation of an intermediate (apo-cPAH(I)). This intermediate is also observed in refolding experiments. Fluorescence studies are consistent with the CD findings. The intermediate apo-cPAH(I) and unfolded state(s) of apo- and holo-cPAH(U) have been characterized by analytical ultracentrifugation (AUC). At 2.4 and 2.8 M GuHCl, 90% of the signal for apo-cPAH has a weight average sedimentation coefficient in water at 20°C (s20,w) of about 48 S, representing multiple aggregate species made of multiple monomers of cPAH. Aggregate formation for apo-cPAH is also confirmed by dynamic light scattering and electron microscopy giving a hydrodynamic radius (R(H)) of 41 nm for apo-cPAH(I) versus 3.5 nm for the native protein.

Tracer-based metabolomics is a systems biology tool that combines advances in tracer methodology for physiological studies, high throughput “-omics” technologies and constraint based modeling of metabolic networks. It is different from the commonly known metabolomics or metabonomics in that it is a targeted approach based on a metabolic network model in cells. Because of its complexity, it is the least understood among the various “-omics”. In this review, the development of concepts and practices of tracer-based metabolomics is traced from the early application of radioactive isotopes in metabolic studies to the recent application of stable isotopes and isotopomer analysis using mass spectrometry; and from the modeling of biochemical reactions using flux analysis to the recent theoretical formulation of the constraint based modeling. How these newer experimental methods and concepts of constraint-based modeling approaches can be applied to metabolic studies is illustrated by examples of studies in determining metabolic responses of cells to pharmacological agents and nutrient environment changes.

Chronic kidney disease (CKD) is associated with accelerated atherosclerosis and cardiovascular disease, which is largely mediated by oxidative stress. We investigated the effect of three glutathione (GSH) precursors: N-acetyl-cysteine (NAC), cystine as the physiological carrier of cysteine in GSH with added selenomethionine (F1), and NAC fortified with selenomethionine (F2) on oxidative stress induced by spermine (a uremic toxin) in cultured human aortic vascular smooth muscle cells (VSMC). VSMC were exposed to spermine (15 μM) with or without the given antioxidants (dose 50, 100, 200, and 500 μg/ml) or vehicle (control) and assessed for intracellular GSH levels, 4-hydroxy-trans-2-nonenal (4-HNE), and incorporation of 13C from glucose into alanine and protein. Spermine exposure reduced intracellular GSH levels, increased 4-HNE, and impaired glucose metabolism through reduction in pyruvate generation and/or transamination. Treatment with NAC had no effect on intracellular glutathione level. In contrast, F1 maintained intracellular GSH at control levels at all four doses. Subsequent studies performed with 200 μg/ml of F1, F2, or NAC (optimal dose) revealed normalization of 4-HNE, whereas restoration of 13C from glucose to alanine or protein to control values was only noted in the F1 group. Spermine-induced alterations in VSMC ultrastructure were prevented in ∼90% of cells treated with F1 but only ∼50% of cells treated with either NAC or F2. In conclusion, F1 was more effective than NAC or F2 in ameliorating spermine-induced reduction in intracellular GSH levels and cellular alterations in VSMC. The cystine-based GSH precursor (F1) is a promising antioxidant, and further studies are needed to examine the effect of this compound in preventing CKD-associated vascular disease.

Ring-necked pheasants (Phasianus colchicus) are able to store dietary calcium as medullary bone, which they may mobilize for future eggshell synthesis. We define this mechanism as calcium-loading. Previous experiments on pheasants conducted to document the importance of calcium in limiting distribution did not account for calcium-loading. We hypothesized that calcium-loading could override experimental calcium treatments of the diet. We measured egg production, egg characteristics, and femoral mineral content for pheasants that were not calcium-loaded on 7 diets differing in calcium from 0.2% to 4.5% and compared these results to a similar study on calcium-loaded pheasants. We predicted that calcium-loaded pheasants would produce more eggs than those that were not calcium-loaded. We also predicted that there would be no significant difference between femur ash fractions in non–calcium-loaded pheasants, but that the ash fraction in calcium-loaded pheasants would differ significantly between the be...

Voltage-gated sodium channels (VGSCs) are responsible for the initiation and propagation of transient depolarizing currents and play a critical role in the electrical signaling between neurons. A null mutation in the VGSC gene SCN8A, which encodes the transmembrane protein Na(v)1.6, was identified previously in a human family. Heterozygous mutation carriers displayed a range of phenotypes, including ataxia, cognitive deficits, and emotional instability. A possible role for SCN8A was also proposed in studies examining the genetic basis of attempted suicide and bipolar disorder. In addition, mice with a Scn8a loss-of-function mutation (Scn8a(med-Tg/+)) show altered anxiety and depression-like phenotypes. Because psychiatric abnormalities are often associated with altered sleep and hormonal patterns, we evaluated heterozygous Scn8a(med-jo/+) mutants for alterations in sleep-wake architecture, diurnal corticosterone levels, and behavior. Compared with their wild-type littermates, Scn8a(med-jo/+) mutants experience more non-rapid eye movement (non-REM) sleep, a chronic impairment of REM sleep generation and quantity, and a lowered and flattened diurnal rhythm of corticosterone levels. No robust differences were observed between mutants and wild-type littermates in locomotor activity or in behavioral paradigms that evaluate anxiety or depression-like phenotypes; however, Scn8a(med-jo/+) mutants did show enhanced spatial memory. This study extends the spectrum of phenotypes associated with mutations in Scn8a and suggests a novel role for altered sodium channel function in human sleep disorders.

Though stress causes complex sleep disruptions that are different in females and males, little is known about how sex influences the ability of stress to alter sleep. To date there have been no comprehensive examinations of whether effects of stress on sleep are sensitive to determinants of sex, such as reproductive hormones. Since restraint stress produces a sexually dimorphic increase in rapid eye movement sleep (REMS) amount in mice that is greater in males than females, in the current study we sought to determine whether estrogens and androgens influence the ability of restraint stress to alter sleep states. We removed the gonads from adult female and male C57BL/6J mice and implanted the mice with recording electrodes to monitor sleep-wake states. Gonadectomized females and males exhibited similar amounts of REMS in response to restraint stress. Mice were then implanted with continuous release hormone pellets. Females received 17beta-estradiol and males received testosterone. Hormone replacement (HR) in females decreased the REMS response to restraint stress while HR in males increased the REMS response to restraint stress. The combined effects of HR in females and males restored the sex difference in the ability of restraint stress to alter REMS. These results demonstrate that sex differences in the effects of stress on REMS are dependent on reproductive hormones and support the view that endogenous or exogenous changes in the reproductive hormone environment influence sleep responses to stress.

In this article we describe the experimental results of a methane hydrate moderator as well as as mesitylene moderator operated at a temperature around 20K at the JESSICA (Julich Experimental Spallation Target Set-up In COSY Area) experiment at the Julich cooler synchrotron COSY. For the first time the cold neutron spectrum of a methane hydrate moderator was experimentally investigated. A comparison with a solid methane and an ice moderator atT = 20 K will be shown. MCNPX simulations with new developedS(α,β) scattering kernels will be compared with experimental data. The applicability of mesitylene and methane hydrate as cold moderators at spallation neutron sources will be discussed.