The interstitial atom of the nitrogenase FeMo-cofactor: ENDOR and ESEEM show it is not an exchangeable nitrogen

A recent high-resolution X-ray crystallographic study (1.16 A) of the Azotobacter vinelandii nitrogenase MoFe protein revealed a previously undetected electron density associated with the active site FeMo-cofactor. The density is located inside the cluster at the center of the "trigonal prism" of six irons and is assigned to a species "X". The identity of species X was not resolved, although the electron density is consistent with a single N, O, or C atom. One proposal is that X is an N atom that derives from and exchanges with N from N2 during catalysis. In the present study, we have examined this possibility by employing 14N and 15N isotopes of N2 along with ENDOR and ESEEM spectroscopies. The WT MoFe protein and alpha-359Arg-->Lys and alpha-381Phe-->Leu variants were allowed to turn over in the presence of 14N2 or 15N2, and then were examined as resting enzymes by ENDOR and ESEEM at X- and Q-bands to look for all 14N and 15N signals coupled to the electron spin of the FeMo-cofactor and to determine if any exchanged during turnover. We have found five peaks in Q-band pulsed ENDOR spectra that appear to arise not only from previously reported N1/N2, which give rise to the ESEEM, but also from one or two additional coupled nitrogens. None of the ENDOR and ESEEM signals vanish or are altered by catalytic turnover with 15N2, and no new 15N signal is detected, leading to the conclusion that if species X is a nitrogen atom, it does not exchange during dinitrogen reduction.     

Manufacturing planning and control: The evolution of MRP and JIT integration

Material Requirements Planning (MRP) and Just-in-Time (JIT) system are directed toward planning and controlling the important characteristics of material flow: how much of what materials flow and when. Since the material flow is at the heart of the manufacturing firm, MRP and JHT are the powerful management tools that could determine the success or failure of an entire manufacturing system. One of the strongest debates in manufacturing has been centered on the performance comparison and compatibility of JIT production system to the existing MRP. The primary intent of this research is to provide an overview of the manufacturing planning and control environment associated with MRP and JIT. Classifying the existing MRP/JIT comparison and integration literature, two different perspectives on MRP/JIT are discussed, and future research area is proposed based on the taxonomy.     

PHASE BEHAVIOR OF DILUTE SODIUM DODECYL SULFATE-HEXANOL-BRINE SYSTEMS

ABSTRACT

Dilute aqueous “solutions” of anionic surfactants injected during enhanced oil recovery processes usually contain liquid crystalline material. Phase behavior has been studied in a model system containing sodium chloride brine with surfactant, alcohol, and salt contents maintained below 6, 10, and 3 weight percent respectively. Pseudoternary phase diagrams at constant salinities are used to represent the results. Some interpretation of the results is given in terms of micellar shape transformations produced by changes in the relative hydrophilic and hydrophobic properties of surfactant-alcohol mixtures.     

Separation and fragmentation study of isocoproporphyrin derivatives by UHPLC‐ESI‐exact mass MS/MS and identification of a new isocoproporphyrin sulfonic acid metabolite

Isocoproporphyrin and its derivatives are commonly used as biomarkers of porphyria cutanea tarda, heavy metal toxicity and hexachlorobenzene (HCB) intoxication in humans and animals. However, most are isobaric with other porphyrins and reference materials are unavailable commercially. The structural characterisation of these porphyrins is important but very little data is available. We report here the separation and characterisation of isocoproporphyrin, deethylisocoproporphyrin, hydroxyisocoproporphyrin and ketoisocoproporphyrin, isolated in the faeces of rats fed with a diet containing HCB, by ultra high performance liquid chromatography‐exact mass tandem mass spectrometry (UHPLC‐MS/MS). Furthermore, we report the identification and characterisation of a previously unreported porphyrin metabolite, isocoproporphyrin sulfonic acid isolated in the rat faeces. The measured mass‐to‐charge ratio (m/z) of the precursor ion was m/z 735.2338, corresponding to a molecular formula of C₃₆H₃₉N₄O₁₁S with an error of 0.3 ppm from the calculated m/z 735.2336. The MS/MS data was consistent with an isocoproporphyrin sulfonic acid structure, derived from dehydroisocoproporphyrinogen by sulfonation of the vinyl group. The metabolite was present in a greater abundance than other isocoproporphyrin derivatives and may be a more useful biomarker for HCB intoxication. Copyright © 2014 John Wiley & Sons, Ltd.     

Training in metabolomics research. I. Designing the experiment,collecting and extracting samples and generating metabolomics data

The study of metabolism has had a long history. Metabolomics, a systems biology discipline representing analysis of known and unknown pathways of metabolism, has grown tremendously over the past 20 years. Because of its comprehensive nature, metabolomics requires careful consideration of the question(s) being asked, the scale needed to answer the question(s), collection and storage of the sample specimens, methods for extraction of the metabolites from biological matrices, the analytical method(s) to be employed and the quality control of the analyses, how collected data are correlated, the statistical methods to determine metabolites undergoing significant change, putative identification of metabolites and the use of stable isotopes to aid in verifying metabolite identity and establishing pathway connections and fluxes. The National Institutes of Health Common Fund Metabolomics Program was established in 2012 to stimulate interest in the approaches and technologies of metabolomics. To deliver one of the program's goals, the University of Alabama at Birmingham has hosted an annual 4‐day short course in metabolomics for faculty, postdoctoral fellows and graduate students from national and international institutions. This paper is the first part of a summary of the training materials presented in the course to be used as a resource for all those embarking on metabolomics research. The complete set of training materials including slide sets and videos can be viewed at http://www.uab.edu/proteomics/metabolomics/workshop/workshop_june_2015.php . Copyright © 2016 John Wiley & Sons, Ltd.     

New constraints on macroscopic compact objects as dark matter candidates from gravitational lensing of type Ia supernovae

We use the distribution, and particularly the skewness, of high redshift type Ia supernovae brightnesses relative to the low redshift sample to constrain the density of macroscopic compact objects (MCOs) in the Universe. The supernova data favor dark matter made of microscopic particles (such as the lightest supersymmetric partner) over MCOs with masses between 10(-2)Mo and 10(10)Mo at 89% confidence. Future data will greatly improve this limit. Combined with other constraints, MCOs larger than one-tenth the mass of Earth (approximately 10(-7)Mo) can be eliminated as the sole constituent of dark matter.     

Differential branching fraction and angular analysis of the decay B0 → K*0 μ+ μ-

The angular distributions and the partial branching fraction of the decay B0 → K*0 μ+ μ- are studied by using an integrated luminosity of 0.37 fb(-1) of data collected with the LHCb detector. The forward-backward asymmetry of the muons, A(FB), the fraction of longitudinal polarization, F(L), and the partial branching fraction dB/dq2 are determined as a function of the dimuon invariant mass. The measurements are in good agreement with the standard model predictions and are the most precise to date. In the dimuon invariant mass squared range 1.00-6.00 GeV2/c4, the results are A(FB)=-0.06(-0.14)(+0.13)±0.04, F(L)=0.55±0.10±0.03, and dB/dq2=(0.42±0.06±0.03)×10(-7) c4/GeV2. In each case, the first error is statistical and the second systematic.     

Search for lepton number violating decays B+ → π- μ+ μ+ and B+ → K- μ+ μ+

A search is performed for the lepton number violating decay B+ → h- μ+ μ+, where h- represents a K- or a π-, using an integrated luminosity of 36 pb(-1) of data collected with the LHCb detector. The decay is forbidden in the standard model but allowed in models with a Majorana neutrino. No signal is observed in either channel and limits of B(B+ → K- μ+ μ+) < 5.4×10(-8) and B(B+ → π- μ+ μ+) < 5.8×10(-8) are set at the 95% confidence level. These improve the previous best limits by factors of 40 and 30, respectively.     

Measurement of the CP-violating phase ϕ(s) in the decay B(s)(0) → J/ψϕ

We present a measurement of the time-dependent CP-violating asymmetry in B(s)(0) → J/ψ? decays, using data collected with the LHCb detector at the LHC. The decay time distribution of B(s)(0) → J/ψ? is characterized by the decay widths Γ(H) and Γ(L) of the heavy and light mass eigenstates, respectively, of the B(s)(0) - B(s)(0) system and by a CP-violating phase ?(s). In a sample of about 8500 B(s)(0) → J/ψ? events isolated from 0.37 fb(-1) of pp collisions at sqrt[s] = 7 TeV, we measure ?(s) = 0.15 ± 0.18(stat) ± 0.06(syst) rad. We also find an average B(s)(0) decay width Γ(s) ≡ (Γ(L) + Γ(H))/2 = 0.657 ± 0.009(stat) ± 0.008(syst) ps(-1) and a decay width difference ΔΓ(s) ≡ Γ(L) - Γ(H) = 0.123 ± 0.029(stat) ± 0.011(syst) ps(-1). Our measurement is insensitive to the transformation (?(s),ΔΓ(s)) ? (π - ?(s), -ΔΓ(s)).