The utilization of the mixtures of monosaccharides by some mucorales

Summary The utilization of mixtures of monosaccharides byBlakeslea trispora Thaxter,Choanephora circinans (Naganish &Kawakami)Hesseltine &Benjamin,Gilbertella persicaria var.indica Mehrotra &Mehrotra andHelicostylum piriforme Bainier was studied. The effect of sorbose on the utilization of other sugars present in the mixtures was also studied. It was found that all the mixtures of sugars in combination with asparagine or ammonium chloride were valueless for all the organisms exceptHelicostylum piriforme. Growth ofHelicostylum piriforme on the mixtures with asparagine as the nitrogen source was better than on the mixtures with ammonium chloride as the source of nitrogen. Asparagine being a favourable source counteractecd sorbose inhibition, while ammonium chloride failed to do so. On the other hand, both of the nitrogen sources failed to counteract sorbose inhibition in the rest of the organisms. None of the organisms could finish sorbose and rhamnose from any of the mixtures within the specified period.     

Studies on active site mutants of P. falciparum adenylosuccinate synthetase: Insights into enzyme catalysis and activation

Adenylosuccinate synthetase catalyzes a reversible reaction utilizing IMP, GTP and aspartate in the presence of Mg²⁺ to form adenylosuccinate, GDP and inorganic phosphate. Comparison of similarly liganded complexes of Plasmodium falciparum, mouse and Escherichia coli AdSS reveals H-bonding interactions involving nonconserved catalytic loop residues (Asn429, Lys62 and Thr307) that are unique to the parasite enzyme. Site-directed mutagenesis has been used to examine the role of these interactions in catalysis and structural organization of P. falciparum adenylosuccinate synthetase (PfAdSS). Mutation of Asn429 to Val, Lys62 to Leu and Thr307 to Val resulted in an increase in K_m values for IMP, GTP and aspartate, respectively along with a 5 fold drop in the k_cat value for N429V mutant suggesting the role of these residues in ligand binding and/or catalysis. We have earlier shown that the glycolytic intermediate, fructose 1,6 bisphosphate, which is an inhibitor of mammalian AdSS is an activator of the parasite enzyme. Enzyme kinetics along with molecular docking suggests a mechanism for activation wherein F16BP seems to be binding to the Asp loop and inducing a conformation that facilitates aspartate binding to the enzyme active site. Like in other AdSS, a conserved arginine residue (Arg155) is involved in dimer crosstalk and interacts with IMP in the active site of the symmetry related subunit of PfAdSS. We also report on the biochemical characterization of the arginine mutants (R155L, R155K and R155A) which suggests that unlike in E. coli AdSS, Arg155 in PfAdSS influences both ligand binding and catalysis.     

Antioxidant Composition of Red and Orange Cultivars of Tomatoes (Solanum lycopersicon L): A Comparative Evaluation

Fourteen commercial cultivars of tomato were analyzed for their antioxidant composition. There was significant difference (p<0.05) in lycopene and phenolic contents between red and yellow cultivars. Red cultivars had higher lycopene content (2.735 to 6.552 mg 100g^?1) than yellow cultivars (0.769 to 1.238 mg 100g^?1). Mean total polyphenolic content and total antioxidant activity in red cultivars was also higher than those in yellow cultivars. Overall cherry tomatoes had highest phenolic content and appeared to be a promising cultivar in terms of their health promoting effects. The results highlight an existing unexploited variability in yellow tomato to improve their antioxidant properties.     

Heterocyclic compounds containing antimony 1. Synthesis, physicochemical properties, crystal and molecular structure of 2-(β-hydroxyethylthio) 1,3,2-oxathiastibolane

The compound (HOCH₂CH₂S) ) (1) has been prepared by the reaction of antimony(III) isopropoxide and 2-mercaptoethanol in a 1:2 molar ratio. Reaction of 1 with MOCH₃ (where M = Li, Na and K) yields bimetallic products of the type, M[(OCH₂CH₂S) )]. All these derivatives have been characterized by elemental analysis, IR, NMR (¹H and ¹³C) spectra and molar conductivity measurements. Crystals of 1 are triclinic, space group P , with a = 6.449(2), b = 10.285(2), c = 13.494(1) Å, α = 78.08(1), β = 75.99(1), γ = 71.54(2)°, V = 815.48 ų, Z = 4, D_calc = 2.239 g cm⁻³, (Mo Kα) λ = 0.7107 Å, μ = 3.55 mm₋₁, F(000) = 528, T = 295 K, final R = 0.0189 for 2344 reflections. One of the two mercaptoethanol moieties in 1 forms a five-membered chelate ring with antimony, Sb(1)---O(11) = 2.023(2) Å and Sb(1)---S(11) = 2.434(1) Å, while the other is bonded through the S atom only, Sb(1)---S(12) = 2.434(1) Å. The angles between these primary bonds with a mean value of 90.2° suggest a basically pyramidal, or pseudo tetrahedral structure if the stereochemically active lone pair is included in the coordination sphere. Two molecules are linked by intermolecular hydrogen bridges. The presence of weak intermolecular secondary bonding, Sb(1)---O(12) = 2.567(3) Å, in the complex indicates that the overall coordination polyhedron is best described in terms of a distorted trigonal bipyramidal arrangement.     

Reduction of Iron by Leaf Extracts and Its Significance for the Assay of Fe(II) Iron in Plants

“Active Fe” in plants has generally been interpreted to be ferrous iron. This study evaluated some of the commonly followed active Fe determination procedures based on Fe(II) measurements. In each of 12 species examined, leaf extracts exhibited strong reducing activity and quickly reduced any added Fe(III) with or without light. The reducing activity was attributed to ascorbic acid and phenols in the plant extracts. The reliability of the Fe(II) iron determination procedures in plants and the interpretation that active Fe is Fe(II) are questionable.     

Peptidolytic monoclonal antibody elicited by a neuropeptide.

We report evidence that a monoclonal antibody raised by immunization with a vasoactive intestinal peptide (VIP)-carrier protein conjugate selectively hydrolyzes VIP and a fluorescence quenched decapeptide (FQ14-22D), representing the region of VIP most susceptible to autoantibody-mediated cleavage (residues 14-22). A high affinity of the antibody for VIP and a lower affinity for FQ14-22D were revealed by kinetic studies and further substantiated by potent inhibition of FQ14-22D cleaving activity by full-length VIP. Sequencing of FQ14-22D hydrolysis products indicated selective cleavage at one peptide bond. These observations suggest that antibodies induced against naturally occurring polypeptide antigens can express peptidolytic activity targeted for specific sequences in the recognition epitope.     

The Nexus of Carbon,Nitrogen, and Biodiversity Impacts from Urban Metabolism

Methodology is developed for linking the urban metabolism (UM) to global environmental stresses on the carbon (C) cycle, nitrogen (N) cycle, and biodiversity loss. UM variables are systematically mapped to the drivers of carbon, nitrogen, and biodiversity impacts. Change in mean species abundance is used as metric of biodiversity loss, by adopting the dose‐response relationships from the GLOBIO model. The main biodiversity drivers related to UM included here are land‐use change (LUC) and atmospheric N deposition. The methodology is demonstrated by studying the nexus for Shanghai in 2006, based on energy and soybean consumption. Results for Shanghai show a strong nexus between C, N, and biodiversity impact due to electricity consumption and energy used in manufacturing industries and construction. Prioritization of the shift away from coal energy will therefore lead to lowering the urban growth impact on all three dimensions. Road transportation, domestic aviation, and the metal industry impact only the C footprint highly, whereas district energy impacts only biodiversity loss highly, showing a weak nexus. Among the global impacts of soybean consumption in Shanghai on biodiversity loss (due to LUC only), the highest impact occurs in Uruguay (0.52%) followed by Brazil (0.05%) and Argentina (0.02%). The local impact on biodiversity loss (i.e., within China) of soybean consumption in Shanghai is 1.03%. However, the methodology and results are limited due to the partial inclusion of drivers, a carbon footprint based on carbon dioxide emissions only, and limitations of biodiversity loss models. Potential to overcome methodological limitations is discussed.     

Initiation of Antiviral B Cell Immunity Relies on Innate Signals from Spatially Positioned NKT Cells

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Polysome shift assay for direct measurement of miRNA inhibition by anti-miRNA drugs

Anti-miRNA (anti-miR) oligonucleotide drugs are being developed to inhibit overactive miRNAs linked to disease. To help facilitate the transition from concept to clinic, new research tools are required. Here we report a novel method—miRNA Polysome Shift Assay (miPSA)—for direct measurement of miRNA engagement by anti-miR, which is more robust than conventional pharmacodynamics using downstream target gene derepression. The method takes advantage of size differences between active and inhibited miRNA complexes. Active miRNAs bind target mRNAs in high molecular weight polysome complexes, while inhibited miRNAs are sterically blocked by anti-miRs from forming this interaction. These two states can be assessed by fractionating tissue or cell lysates using differential ultracentrifugation through sucrose gradients. Accordingly, anti-miR treatment causes a specific shift of cognate miRNA from heavy to light density fractions. The magnitude of this shift is dose-responsive and maintains a linear relationship with downstream target gene derepression while providing a substantially higher dynamic window for aiding drug discovery. In contrast, we found that the commonly used ‘RT-interference’ approach, which assumes that inhibited miRNA is undetectable by RT-qPCR, can yield unreliable results that poorly reflect the binding stoichiometry of anti-miR to miRNA. We also demonstrate that the miPSA has additional utility in assessing anti-miR cross-reactivity with miRNAs sharing similar seed sequences.