Release of monomeric sugars from Miscanthus sinensis by microwave-assisted ammonia and phosphoric acid treatments

Microwave-assisted ammonium hydroxide (NH₄OH) followed by phosphoric acid (H₃PO₄) treatments were used to release monomeric sugars from Miscanthus sinensis grown in Cha-Chueng-Sao province, Thailand. Treatment with 1.0% (w/v) NH₄OH, 15:1 liquid-to-solid ratio (LSR) at 120 °C temperature for 15 min liberated 2.9 g of monomeric sugars per 100 g of dried biomass, whereas the corresponding yield for a treatment with 1.78% v/v H₃PO₄, 15:1 LSR at 140 °C for 30 min was 62.3 g/100 g. The two-stage pretreatment, treatment with NH₄OH at 120 °C temperature for 15 min followed by treatment with H₃PO₄ at 140 °C for 30 min, impressively provided the highest total monomeric sugar yield of 71.6 g/100 g dried biomass.     

Highly hydroxylated or γ-cyclodextrin-bicapped water-soluble derivative of fullerene: The antioxidant ability assessed by electron spin resonance method and β-carotene bleaching assay

Antioxidant ability of the water-soluble derivative of fullerene (C60), prepared by high-degree hydroxylation [C60-(OH)₃₂·8H₂O] or C60/γ-cyclodextrin (1:2 mol/mol) clathrate formation [C60/(γ-CD)₂], was assessed by electron spin resonance method and β-carotene bleaching assay. These C60 derivatives have an ability to diminish a 1:2:2:1 quartet ESR spectrum attributed to hydroxyl radicals (OH) as shown by DMPO-spin trap/ESR method. Meanwhile, a singlet radical-signal different from OH-attributed signals increased in a manner dependent on concentrations of C60-(OH)₃₂·8H₂O. This might suggest that C60-(OH)₃₂·8H₂O scavenges OH owing to dehydrogenation of C60-(OH)₃₂·8H₂O, and is simultaneously oxidized to a stable radical species, which may be a dehydrogenated fullerenol radical (C60-O). Furthermore, these water-soluble derivatives of C60 suppressed fading of yellowish color characteristic of intact β-carotene in β-carotene bleaching assay. Antioxidant abilities of these derivatives were assessed as retention of yellowish color (viz absorbance at 470 nm) for 180 min. Namely, β-carotene-attributed chromaticity (% relative absorbance at 470 nm compared with the control) after 180 min was 69% for C60-(OH)₃₂·8H₂O (400 μM: C60-eq.), and 32% for C60/(γ-CD)₂ (400 μM: C60-eq.), whereas it was 6% for l(+)-ascorbic acid (400 μM) which is hydrophilic, and 85% for (±)-α-tocopherol (400 μM) which is lipophilic, respectively. Thus C60-(OH)₃₂·8H₂O and C60/(γ-CD)₂ can scavenge OH, and have a distinct antioxidative activity in the aqueous system containing linoleic acid which is abundantly contained in the cell membrane together with other unsaturated lipids. These C60 derivatives have a potential to protect the cell membrane from oxidative stress due to OH.     

HPLC determination of tryptophan in foodstuffs using barium hydroxide hydrolysis at 140°C

Summary Hydrolysis of samples of food and feedstuffs with 2.7 N Ba(OH)₂ at 140°C for 4 h was tested for the recovery of tryptophan. On the basis of 100% yield for the tryptophan content, corresponding to samples determined after 16h-hydrolysis at 125°C, the recovery averaged 98.7 ± 0.9% SD or 99.4 ± 2% depending on how the bulk or aliquot of hydrolysate was analysed (conventional or simplified procedure). Tryptophan can be assayed by high performance liquid chromatography and fluorescence detection within 8h, 4h-hydrolysis at 140°C and a simplified procedure.     

Investigation of γE-crystallin target protein binding to bovine lens alpha-crystallin by small-angle neutron scattering

α-Crystallin, one of the main constituent proteins in the crystalline lens, is an important molecular chaperone both within and outside the lens. Presently, the structural relationship between α-crystallin and its target proteins during chaperone action is poorly understood. It has been hypothesised that target proteins bind within a central cavity. Small-angle neutron-scattering (SANS) experiments in conjunction with isotopic substitution were undertaken to investigate the interaction of a target lens protein (γE-crystallin) with α-crystallin (α_H) and to measure the radius of gyration (Rg) of the proteins and their binary complexes in solution under thermal stress. The size of the α_H in D₂O incubated at 65 °C increased from 69 ± 3 to 81 ± 5 Å over 40 min, in good agreement with previously published small-angle X-ray scattering (SAXS) and SANS measurements. Deuterated γE-crystallin in H₂O buffer (γE_D/H₂O) and hydrogenous γE-crystallin in D₂O buffer (γE_H/D₂O) free in solution were of insufficient size and/or too dilute to provide any measurable scattering over the angular range used, which was selected primarily to investigate γE:α_H complexes. The evolution of the aggregation size/shape as an indicator of α_H chaperone action was monitored by recording the neutron scattering in different H:D solvent contrasts under thermally stressed conditions (65 °C) for binary mixtures of α_H, γE_H, and γE_D. It was found that Rg(α_H:γE_D/D₂O) > Rg(α_H:γE_H/D₂O) > Rg(α_H/D₂O) and that Rg(α_H:γE_H/D₂O) ≈ Rg(α_H/D₂O). The relative sizes observed for the complexes weighted by the respective scattering powers of the various components imply that γE-crystallin binds in a central cavity of the α-crystallin oligomer, during chaperone action.     

Optimization of reaction variables for sucrose monoester production using lipase in a solvent free system by Taguchi's method

Taguchi's method was attempted successfully to optimize the reaction variables and their ranges for sucrose monoester production. Optimal conditions determined by Taguchi's method were 35°C, 200rpm, capric acid (40mmol) and sucrose (3mmol), 24h, 0.5g lipase, 0.72g Ba(OH)2 ·8H2O, 0.72g Ba(OH)2·1H2O. A product yield of 21% was obtained. © Rapid Science Ltd. 1998     

Comparative study of alkali-soluble hemicelluloses isolated from bamboo (Bambusa rigida)

The physicochemical properties and structural characteristics of seven alkali-soluble hemicellulosic preparations were determined. These were extracted from bamboo (Bambusa rigida) with 1 M NaOH, KOH, LiOH, NH₃·H₂O, (CH₃CH₂)₃N, Ca(OH)₂, Ba(OH)₂, respectively, at 50 °C for 3 h, were comparatively studied. Sugar analysis showed that these hemicelluloses contained d-xylose as the major constituent, along with d-glucose and l-arabinose in noticeable amounts. Uronic acids, principally 4-O-methyl-d-glucuronic acid, occurred in a small amount. Furthermore, based on the sugar analysis and FTIR and NMR spectroscopy, it can be concluded that the hemicelluloses consist of a backbone of β-(1→4)-linked d-xylopyranosyl units having branches of arabinose and 4-O-methyl-d-glucuronic acid. Nitrobenzene oxidation revealed that the hemicelluloses obtained are mostly free of bound lignins. Moreover, it is noteworthy that hemicelluloses isolated with the different alkaline solutions presented different chemical compositions and slightly dissimilar structural features, indicating that alkalinity played an important role in cleaving the chemical linkages between the hemicelluloses and the lignins.     

Crystal structures and spectroscopic properties of palladium complexes isolated from Pd-EDTA solutions

Two complexes were isolated from aqueous Pd(NO₃)₂-Na₂H₂EDTA solutions and studied by single crystal X-ray diffraction, IR/Raman spectroscopy, and photoelectron spectroscopy. The first complex, Pd(Hkpda)₂ (kpda = ketopiperazinediacetate dianion, C₈H₁₀N₂O₅), forms yellow parallelepipeds during slow evaporation of Pd(NO₃)₂-Na₂H₂EDTA solution at 25 °C, and is a result of EDTA oxidation. The second one, [Pd(μ-H₂EDTA)]₂ · 2NaNO₃ · 7.5H₂O, with two EDTA molecules acting as bridges between two palladium atoms, forms yellow bipyramides during fast evaporation at 60 °C. In both complexes, the palladium atoms adopt a planar trans-geometry by bonding to two nitrogen and two oxygen atoms of two ligand molecules.     

Synthesis, chemical speciation and SOD mimic assays of tricarballylic acid-copper(II) and imidazole-tricarballylic acid-copper(II) complexes

The coordination behavior of copper(II) with tricarballylic acid (H₃TCA) and imidazole (Imz) is described. Speciation in aqueous solution has been determined at 25 °C and 0.15 M NaCl ionic strength by potentiometric measurements and EPR characterization of the species. Two new compounds CuTCAH · 3H₂O and CuTCAHImz · 2H₂O were obtained and characterized by elemental analysis diffuse reflectance, FTIR (Fourier transform infrared spectroscopy), EPR and thermal behavior. Their in vitro superoxide dismutase-mimetic activities have been tested.     

Involvement of a Ca2+-calmodulin interaction in the yeast-mycelial (Y-M) transition of Candida albicans

A yeast-mycelium (Y-M) transition of Candida albicans (3153A) was induced by 1.5 mM CaCl₂ · 2H₂O in defined liquid medium, pH 7, at 25 °C. Germ tube formation was detected after approximately 8 h and peaks of maximum germination occurred at approximately 20 h in all experimental treatments. Non-toxic concentrations of the calmodulin inhibitor R24571 almost completely suppressed germ tube formation whereas trifluoperazine (TFP) and the Ca²⁺ ionophore A23187 were only about half as effective. Further Ca²⁺ addition failed to reverse the inhibitory effect of R24571 and induced only about 10% of the cells inhibited by TFP or A23187 to germinate.     

DNA loss during C-banding of chromosomes of Lilium longiflorum

Root tip chromosomes were uniformly labelled with ³H-thymidine and replicate squashes were made. One set was untreated, one incubated in Ba(OH)₂ solution, and a further set treated sequentially in Ba(OH)₂ and hot saline-citrate (2 × SSC) to reveal C-bands. All replicates were autoradiographed and comparative grain counts made. Differences in grain numbers per metaphase cell showed that Ba(OH)₂ extracted 40% of label, and that a further 23% was lost in the subsequent SSC incubation. The distribution of grains was mapped along a sample of each of five individually-recognisable chromosomes at the three treatment stages. Within each chromosome, the number of grains per segment did not differ significantly from a random distribution. This was true for all five chromosomes at all three stages of treatment, whether or not the regions were C-banded. — We conclude that DNA extraction occurs progressively during C-banding in Lilium, but that C-bands are not dark because of their relatively high retention of DNA.     

Treatment of cotton seeds for germination

Summary Effects of H₂SO₄, H₂O₂, C₂H₅OH and of acetone on germination of cotton seeds and on the growth of seedling root were studied under laboratory conditions. H₂SO₄ or H₂O₂ hastened germination but decreased the ultimate number of seeds germinated. Higher temperature (27°C) hastened germination and increased the number of seeds germinated. Root length remained unaffected by treatments at room temperature while H₂SO₄ or H₂SO₄/H₂O₂ substantially decreased it at 27°C.     

Novel supramolecular compounds based on Cucurbit[6]uril, 1,8-diaminooctane and octahedral thiohydroxo anions with cluster core [Re6S8]

Supramolecular compounds {C₈N₂H₂₂@Cuc[6]}{Re₆S₈(H₂O)₂(OH)₄}·18H₂O (1), and K₂{C₈N₂H₂₂@Cuc[6]}{Re₆S₈(OH)₆}·14H₂O (2) were obtained by crystallization from aqueous solutions that contained the macrocyclic cavitand cucurbit[6]uril (C₃₆H₃₆N₂₄O₁₂), 1,8-diaminooctane and the cluster thiohydroxo complex [Re₆S₈(OH)₆]⁴⁻. The resultant composition of the formed compounds depends on the experiment technique. According to the X-ray diffraction analysis, 1,8-diaminooctane molecules are encapsulated within the cavity of the cucurbit[6]uril molecules in such a way that the aminogroups are above and below the plane of the cavitand. The 1,8-diaminooctane molecules formed hydrogen bonds with the cavitand and the cluster thiohydroxo complexes to give chains.     

A stable second-order NLO and luminescent Cd(II) complex based on 6-hydroxynicotinic acid

Employing an unsymmetrical 6-hydroxynicotinic acid as a bridging ligand, we have prepared a new NLO-active two-dimensional cadmium coordination polymer [Cd₈(nicO)₈(phen)₈ · (H₂O)]_n (1) (H₂nicO = 6-hydroxynicotinic acid, phen = 1,10-phenanthroline) under hydrothermal condition. At room temperature, 1 shows strong fluorescent emission bands in the solid state and a SHG efficiency of about 8 times that of KDP (KH₂PO₄). On the basis of TG/DTA analyses, the structure is thermally stable up to 370 °C.     

Reaction of hydrogen peroxide with hexaaquacobalt(III) perchlorate

The reaction of with H₂O₂ in 1.0 M HClO₄/LiClO₄ was found to be first-order in both reactants and the [H⁺] dependence of the second-order rate constant is given by k_2obs = b/[H⁺], b at 25 °C is 26.4 ± 0.5 s⁻¹. The rate law shows a simple inverse dependence on [H⁺] that is consistent with a rapidly maintained equilibrium between and its hydrolyzed form Co(H₂O)₅(OH)²⁺, followed by the rate controlling step, i.e. oxidation of H₂O₂ by Co(H₂O)₅(OH)²⁺.     

Preparation of planar chiral ferrocenes containing a fluorous alcohol function

Ferrocene reacts with hexafluoroacetone trihydrate in refluxing octane to afford >80% yields of [CpFe(η⁵-C₅H₄C(CF₃)₂OH)] (X-ray), carrying out the reactions at 180 °C gives an additional 5% yield of [Fe(η⁵-C₅H₄C(CF₃)₂OH)₂] (X-ray).The mono alcohol is lithiated with Bu^tOK/BuⁿLi/TMEDA affording partial conversion to mixtures of [CpFe(1,2-η⁵-C₅H₃C(CF₃)₂OH)(X)] and [Fe(η⁵-C₅H₄X)(1,2-η⁵-C₅H₃C(CF₃)₂OH)(X)] (X = SMe, CPh₂OH) upon reaction with Me₂S₂ or OCPh₂.For X = CPh₂OH both structures are crystallographically characterised.Enantiopure [CpFe(1,2-η⁵-C₅H₃C(CF₃)₂OH)(SMe)] can be prepared from (R)-[CpFe(η⁵-C₅H₄S(O)C₆H₄Me)] via [CpFe(1,2-η⁵-C₅H₃S(O)C₆H₄Me)(C(CF₃)₂OH)] (X-ray) or [CpFe(1,2-η⁵-C₅H₃S(O)C₆H₄Me)(SMe)].Related procedures allow the preparation of [CpFe(1,2-η⁵-C₅H₃CPh₂OH)(Y)] (Y = SMe, CHO (X-ray), C(CF₃)₂OH) and[CpFe(1,2-η⁵-C₅H₃C(CF₃)₂OH)(CHO)].     

Crystallographic study of lead-substituted hydroxyapatite synthesized by high-temperature mixing method under hydrothermal conditions

The solid solutions in the system of Ca and Pb hydroxyapatite, Ca_10−χPb_χHA (χ = 0-10), were successfully synthesized by high-temperature mixing method (HTMM) at 200 °C for 12 h under hydrothermal conditions. The samples were characterized by X-ray diffraction, chemical analysis and electron microscopic observation, and the site of the metal ions in the solid solutions was analyzed with the Rietveld method. The lattice parameters of the solid solutions prepared by both of the methods didn’t vary linearly with Pb contents, and they have two turning points of 0.4 and 0.6 of Pb content. It was found that Pb ions in the solid solutions preferentially occupied the M (2) site in the apatite structure. HTMM gives Ca-Pb HA solid solutions much better crystallization. However, due to the formation of intermediate compound of Pb₃O₂(OH)₂ in the Pb(NO₃)₂·4H₂O solution before mixing with (NH₄)₂HPO₄ solution at 200 °C, HTMM causes the decrease of crystallization of the samples with high Pb content.     

Content of iron and copper in the nuclei and induction of pH 9-labile lesions in L5178Y sublines inversely cross-sensitive to H2O2 and x-rays

Cells from the L5178Y murine lymphoma subline LY-R are twice as resistent to killing by ionizing radiation than the subline LY-S. In contrast, LY-R cells are more sensitive to killing by H₂O₂, the effect being more pronounced at 37 °C than 0 °C. Initial DNA damage after H₂O₂ treatment (both temperatures, 5 min) has been estimated by the comet assay (single-cell gel electrophoresis) and fluorescent halo technique. According to both methods, the initial damage is significantly higher in LY-R cells, particularly that inflicted at O °C. Differences between DNA unwinding and rewinding abilities at pH 9 and 6.9 (estimated by the fluorescent halo technique) point to a considerable difference in pH-9-labile damage between the sublines, as observed previously for x-irradiated cells (Kapiszewska et al. 1992). In contrast to findings with x-irradiated cells, however, after H₂O₂ treatment this damage is more extensive in LY-R cells than in LY-S cells. Thus, the initial pH-9-labile damage corresponds to the pattern of sensitivity to H₂O₂ and x-rays. We suggest that this is caused by different proportions of cuprous and ferric ions found in the nuclei of LY sublines and by the different ability of these ions to react with H₂O₂ and water radiolysis products. The copper/iron ratio in the nucleus is 1.31 in LY-R cells and 4.84 in LY-S cells.     

Thermodynamic and spectroscopic studies of cadmium(II)-N-(phosphonomethyl)glycine (PMG) complexes

Speciation and equilibria in the H⁺-Cd²⁺-N-(phosphonomethyl)glycine (PMG, H₃L) system have been studied in 0.1 M Na(Cl) medium at 25.0 °C. Formation constants for a series of mononuclear complexes, CdHL, CdL⁻, CdL₂ ⁴⁻ and CdL(OH)²⁻ were determined from potentiometric titrations. The structures of the predominating species CdL⁻, and CdL₂ ⁴⁻ in solution were investigated using EXAFS and IR spectroscopic techniques. In the 1:1 complex bonds are formed between the Cd(II) ion and all three donor groups (amino, carboxylate, phosphonate) of the PMG molecule resulting in two 5-membered chelate rings. At the remaining three of the corners of the distorted Cd(II) octahedra oxygens were found which are replaced by donor groups of a second PMG molecule in the 1:2 complex. Furthermore, a solid phase consisting of Cd₉(PMG)₆(H₂O)₁₂ · 6H₂O crystals was synthesized and the crystal structure was determined. The structure consists of six CdL⁻ octahedra connected through a seventh Cd-O octahedron in the centre of the entity, with two additional Cd-O octahedra located at the apices of the unit formed.     

The cellulase activity of an extreme thermophile

Summary The carboxymethylcellulase activity concentrated from the extremely thermophilic anaerobe H173 was found to have a pH optimum of 6.5–7.0. The enzyme activity was stabilised by the addition of dithiothreitol and CaCl₂·2H₂O and was very stable at 80° C, retaining 77% of the initial activity after 120 min incubtation. At min and after 120 min only 3% of the initial activity remained. With the enzyme dissolved in buffer, glucose and cellobiose were formed from the hydrolosis of Avicel. In culture medium the Avicel-solubilising activity was insensitive to the presence of up tp 50 mm glucose and showed linear glucose accumulation over a period of days at 70° C. HPLC analysis established that glucose was the major end-product of hydrolysis in the culture broths.Offprint requests to: H. W. Morgan     

The temperature dependence of the acute toxicity of copper to a freshwater pond snail,Viviparus bengalensis L.

Static bioassays with copper (as CuSO₄ · 5H₂O) were conducted in laboratory with a freshwater pond snailViviparus bengalensis, at different environmental temperatures. The 96 hr LC50 values in ppm of Cu were 0.060 at 32.5 °C; 0.066 at 24 °C; 0.09 at 27.3 °C and 0.39 at 20.3 °C. In higher copper concentrations some behavioural changes such as secretion of mucus, discharge of eggs and embryos were noted. The results indicate that toxicity to copper increases with temperature increase.