Chemical and structural characterization of α-N-acetylgalactosaminidase I and II from starfish, <Emphasis Type="Italic">asterina amurensis</Emphasis>

Background

The marine invertebrate starfish was found to contain a novel α-N-acetylgalactosaminidase, α-GalNAcase II, which catalyzes removal of terminal α-N-acetylgalactosamine (α-GalNAc), in addition to a typical α-N-acetylgalactosaminidase, α-GalNAcase I, which catalyzes removal of terminal α-N-acetylgalactosamine (α-GalNAc) and, to a lesser extent, galactose. The interrelationship between α-GalNAcase I and α-GalNAcase II and the molecular basis of their differences in substrate specificity remain unknown.

Results

Chemical and structural comparisons between α-GalNAcase I and II using immunostaining, N-terminal amino acid sequencing and peptide analysis showed high homology to each other and also to other glycoside hydrolase family (GHF) 27 members. The amino acid sequence of peptides showed conserved residues at the active site as seen in typical α-GalNAcase. Some substitutions of conserved amino acid residues were found in α-GalNAcase II that were located near catalytic site. Among them G171 and A173, in place of C171 and W173, respectively in α-GalNAcase were identified to be responsible for lacking intrinsic α-galactosidase activity of α-GalNAcase II. Chemical modifications supported the presence of serine, aspartate and tryptophan as active site residues. Two tryptophan residues (W16 and W173) were involved in α-galactosidase activity, and one (W16) of them was involved in α-GalNAcase activity.

Conclusions

The results suggested that α-GalNAcase I and II are closely related with respect to primary and higher order structure and that their structural differences are responsible for difference in substrate specificities.

     

Sorption behavior of Sn(II) onto Haro river sand from aqueous acidic solutions

The sorption behavior of Sn(II) onto Haro river sand has been examined with respect to nature of electrolyte, agitation time, dosage of sorbent and concentration of sorbate. Maximum sorption (95.5%) has been achieved from 0.034M hydrochloric acid solution after equilibrating sorbate (2·10⁻⁵M) and sorbent (50 mg) for 120 minutes at aV/W ratio of 90 cm³·g⁻¹. The kinetic data have been subjected to Morris-Weber and Lagergren equations. The kinetics of sorption proceeds a two stage process consisting of a relatively slow initial uptake followed by a much rapid increase in the sorption. The rate constant of intraparticle transport, K_d, comes out to be 8.75·10⁻⁸ mol·g⁻¹·min^−1/2 and the first order rate constant for sorption is 0.0416 min⁻¹. The sorption data of Sn(II) onto Haro river sand followed Langmuir, Freundlich and Dubinin-Radushkevich (D-R) type isotherms. The Langmuir constant,Q, related to sorption capacity and,b, related to sorption energy are computed to be 10.6±1.1 μmol·g⁻¹ and 1123±137 dm³·mol⁻¹, respectively. The D-R isotherm yields the values ofC _m=348±151 μmol·g⁻¹ and β=−0.01044±0.0008 mol²·kJ⁻² and ofE=6.9±0.3 kJ·mol⁻¹. In all three isotherms correlation factor (γ) is ≥0.99. The influence of common anions and cations on the sorption has been investigated. Zn(II), Mg(II), oxalate, Pb(II), Mn(II) and tartrate reduce the sorption significantly whereas Fe(II) causes substantial increase in the sorption. It is essential that all ions causing a decrease in the sorption of Sn(II) must be absent from the sorptive solution otherwise low sorption yields would result.     

Quantitative estimation of magnesite by differential thermal analysis

A method for quantitative estimation of magnesite in magnesite-rich rocks by differential thermal analysis, using magnesium hydroxide as internal standard, is presented. The effects of variations due to particle size and impurities are discussed.     

Radiometric microbiological estimation of vitamin B12

Estimation of vitamin B₁₂ in blood is very important to determine in decifiency and diagnosis of anemic patients. Vitamin B₁₂ in blood can be estimated by spectrochemical, enzymatic, radioisotopic and microbiological methods. In the present study vitamin B₁₂ was determined in 48 normal subjects of Rawalpindi/Islamabad by radiometric microbiological assay (RMA) technique using a very rapid, sensitive and automated instrument Bactec 460. In this procedure¹⁴C-glucose media and microorganismsLactobacillus leichmannii were used. The sensitivity of the method for vitamin B₁₂ is 1 pg/ml and the vitamin B₁₂ found in normal subjects was in the range of 105–535 pg/ml with a median value of 246±6 pg/ml.     

DFT study of boron trichloride adsorption on the surface of Al12N12 nanocluster

In the present study, ability of Al₁₂N₁₂ nanocluster as a new adsorbent for boron trichloride is studied by using density functional theory (DFT) calculations. Two distinct relaxed geometries of Al₁₂N₁₂-BCl₃ complex are located. The dominant configuration (B) involves chemisorption of BCl₃ on aluminum nitride nanocluster with an adsorption energy of ?303 kJ/mol compared to ?33 kJ/mol for a less favourable configuration A (based on wB97XD functional). The enthalpy and Gibbs free energy of interaction for each relaxed was also analysed for better understanding of the thermochemistry of adsorption process. The charge transfer analysis reveals opposing trend in both configurations. Configuration A showed transfer of charge from BCl₃ to nanocluster (+0.123 e), whereas reverse direction of charge transfer was found for the other configuration (?0.083 e). Furthermore, the frontier molecular orbital as well as densities of states of all systems are analysed to follow the changes in the electronic structure of Al₁₂N₁₂ upon adsorption of BCl₃.     

Mössbauer Studies of Core-Shell FeO/Fe3O4 Nanoparticles

Physics of the Solid State - FeO/Fe3O4 nanoparticles were synthesized by thermal decomposition. Electron microscopy revealed that these nanoparticles were of the core-shell type and had a spherical...     

Synthesis of Symmetric Bridged Bis-Pyrazolone Based Metal Complex Acid Dyes and their Applications on Leather

Our present investigation aims at the synthesis and application of new, symmetric bridged bis-pyrazolone based acid dyes. The bis-pyrazolone compounds were accomplished from bis- hydrazine of 4,4′-Diaminostilbene-2,2′-disulfonic acid and ethyl acetoacetate. The bis-pyrazolones have been coupled with diazonium salts of o-hydroxyl aromatic amines which resulted in ligand dyes. The intermediate ligand dyes were treated with 3d transition metals to achieve the targeted metal complex acid dyes. The structures of investigated compounds were confirmed with the help of spectroscopic techniques. Dyes were applied on leather and their application parameters including their light fastness, wash fastness and rubbing fastness were determined.

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Graphical Abstract Symmetric brymmetric Bridged bis-Pyrazolone based Metal Complex Acid

     

Direct observation of diffuse cone behavior in de Vries smectic-A and -C phases of organosiloxane mesogens

Simultaneous and direct x-ray measurements of the smectic layer spacing, molecular tilt, and orientational order in the de Vries smectic A (SmA) and C (SmC) phases of two organosiloxane mesogens reveal that (i) the SmC (tilt) order parameter exponent β=0.26±0.01 for 2nd order SmA-SmC transition--in excellent agreement with the tricritical behavior, (ii) the siloxane and hydrocarbon parts of the molecules are segregated and oriented parallel to the director with very different degree of orientational order, and (iii) thermal evolution of the effective molecular length is different in the two phases.     

Antiangiogenic Activity and Bioassay‐guided Isolation of Aqueous Extract of Orthosiphon stamineus

Orthosiphon stamineus Benth. (Family: Lamiaceae), a famous traditional medicinal plant in tropical and sub‐tropical region, is being used for treating various ailments including inflammatory conditions. Since anti‐inflammation and antiangiogenesis involve cyclooxygenase‐II pathway, the present study is aimed to carry out antiangiogenic studies and bioassay‐guided isolation on aqueous extract of the plant. After antiangiogenic activity studies on the extract, fractions were prepared by partitioning the extract with hexane, chloroform and ethyl acetate, which were then evaluated for antiangiogenic activity. The most active, hexane fraction (HF) upon activity‐guided isolation gave three compounds, 1 (betulinic acid), 2 (oleanolic acid) and 3 (ursolic acid). The total contents of these three compounds were determined in the extract and its fractions using HPLC. The extract and HF exhibited 20% and 80% antiangiogenic activity, respectively, whereas, chloroform and ethyl acetate fractions were inactive. The three isolated compounds exhibited 100% antiangiogenic activity. Median effective concentrations (EC_50s) of HF, 1, 2 and 3 were found to be 84, 32, 34 and 38 μg/mL, respectively. The results of this study indicate that hexane fraction and the isolated compounds have promising antiangiogenic activity. Additionally, the isolated compounds may be sued as analytical markers to standardize extracts and formulations prepared from O. stamineus.