Use of mycelial waste from the drug industry

It was shown that mycelial waste (MW) from manufacture of riboxin and neomycin was useful as an active filler for press compositions and a raw material for fuels. Addition of MW to a polymer press composition, depending on the amount added, provided a 2-10-fold decrease in the time of gel formation in the mixture and hardening of the products as well as a significant decrease in their shrinkage during the hardening. Optimal physicomechanical properties were achieved when the contents of MW and polymer in the press composition were equal. Pyrolysis of dry MW at a temperature of 450-500 degrees C resulted in formation of a product which was not inferior to the known fuels in its physicochemical properties. The low freezing point of the prepared fuel as compared to that of oil fuels allowed one to recommend it as a low-temperature additive to the fuels lowering their freezing points.     

Development of a high-temperature air-blown gasification system

Current status of high-temperature air-blown gasification technology development is reviewed. This advanced gasification system utilizes preheated air to convert coal and waste-derived fuels into synthetic fuel gas and value-added byproducts. A series of demonstrated, independent technologies are combined to form the core of this gasification system. A high-temperature, rapid devolatilization process is used to enhance the volatile yields from the fuel and to improve the gasification efficiency. A high-temperature pebble bed filter is used to remove to the slag and particulates from the synthetic fuel gas. Finally, a novel regenerative heater is used to supply the high-temperature air for the gasifier. Component development tests have shown that higher gasification efficiencies can be obtained at more fuel-rich operating conditions when high-temperature air is used as the gasification agent. Test results also demonstrated the flex-fuel capabilities of the gasifier design. Potential uses of this technology range from large-scale integrated gasification power plants to small-scale waste-to-energy applications.     

Cyclic AMP-dependent signaling system is a primary metabolic target for non-thermal effect of microwaves on heart muscle hydration

Previously, we have suggested that cell hydration is a universal and extra-sensitive sensor for the structural changes of cell aqua medium caused by the impact of weak chemical and physical factors. The aim of present work is to elucidate the nature of the metabolic messenger through which physiological solution (PS) treated by non-thermal (NT) microwaves (MW) could modulate heart muscle hydration of rats. For this purpose, the effects of NT MW–treated PS on heart muscle hydration, [³H]-ouabain binding with cell membrane, ⁴⁵Ca²⁺ uptake and intracellular cyclic nucleotides contents in vivo and in vitro experiments were studied. It is shown that intraperitoneal injections of both Sham-treated PS and NT MW–treated PS elevate heart muscle hydration. However, the effect of NT MW–treated PS on muscle hydration is more pronounced than the effect of Sham-treated PS. In vitro experiments NT MW–treated PS has dehydration effect on muscle, which is not changed by decreasing Na⁺ gradients on membrane. Intraperitoneal injection of Sham- and NT MW–treated PS containing ⁴⁵Ca²⁺ have similar dehydration effect on muscle, while NT MW–treated PS has activation effect on Na⁺/Ca²⁺ exchange in reverse mode. The intraperitoneal injection of NT MW–treated PS depresses [³H]-ouabain binding with its high-affinity membrane receptors, elevates intracellular cAMP and decreases cGMP contents. Based on the obtained data, it is suggested that cAMP-dependent signaling system serves as a primary metabolic target for NT MW effect on heart muscle hydration.     

Biomass–oxygen gasification in a high-temperature entrained-flow gasifier

The technology associated with indirect biomass liquefaction is currently arousing increased attention, as it could ensure a supply of transportation fuels and reduce the use of petroleum. The characteristics of biomass–oxygen gasification in a bench-scale laminar entrained-flow gasifier were studied in the paper. Experiments were carried out to investigate the influence of some key factors, including reaction temperature, residence time and oxygen/biomass ratio, on the gasification. The results indicated that higher temperature favored H₂ and CO production. Cold gas efficiency was improved by > 10% when the temperature was increased from 1000 to 1400 °C. The carbon conversion increased and the syngas quality was improved with increasing residence time. A shorter residence resulted in incomplete gasification. An optimal residence time of 1.6 s was identified in this study. The introduction of oxygen to the gasifier strengthened the gasification and improved the carbon conversion, but lowered the lower heating value and the H₂/CO ratio of the syngas. The optimal oxygen/biomass ratio in this study was 0.4. The results of this study will help to improve our understanding of syngas production by biomass high-temperature gasification.     

Fermentation of biomass-generated synthesis gas: effects of nitric oxide

The production of renewable fuels, such as ethanol, has been steadily increasing owing to the need for a reduced dependency on fossil fuels. It was demonstrated previously that biomass-generated synthesis gas (biomass-syngas) can be converted to ethanol and acetic acid using a microbial catalyst. The biomass-syngas (primarily CO, CO(2), H(2), and N(2)) was generated in a fluidized-bed gasifier and used as a substrate for Clostridium carboxidivorans P7(T). Results showed that the cells stopped consuming H(2) when exposed to biomass-syngas, thus indicating that there was an inhibition of the hydrogenase enzyme due to some biomass-syngas contaminant. It was hypothesized that nitric oxide (NO) detected in the biomass-syngas could be the possible cause of this inhibition. The specific activity of hydrogenase was monitored with time under varying concentrations of H(2) and NO. Results indicated that NO (at gas concentrations above 40 ppm) was a non-competitive inhibitor of hydrogenase activity, although the loss of hydrogenase activity was reversible. In addition, NO also affected the cell growth and increased the amount of ethanol produced. A kinetic model of hydrogenase activity with inhibition by NO was demonstrated with results suggesting there are multiple binding sites of NO on the hydrogenase enzyme. Since other syngas-fermenting organisms utilize the same metabolic pathways, this study estimates that NO < 40 ppm can be tolerated by cells in a syngas-fermentation system without compromising the hydrogenase activity, cell growth, and product distribution.     

Effects of the reforming reagents and fuel species on tar reforming reaction

In this study, using wood chips and polyethylene (PE) as fuels, the effects of air and/or steam as reagents on the tar reforming were clarified quantitatively with a simulated gasifier/reformer apparatus of a two-staged gasification process. The results show that when only steam or air was supplied into the reformer, the tar residual rate (defined as the ratio of the tar amount in the reformed gas to the tar amount in the pyrolysis gas) and the carbon particulate concentration in both reformed gases produced from pyrolysis gases of wood chips and PE decreased with the increase of the steam ratio (H₂O/C, 0–1.0) or the air ratio (ER, 0–0.30). Supplying steam into the reformer to suppress carbon particulate formation for PE pyrolysis gas is more effective than for wood chips pyrolysis gas. Comparing with the results of steam only reforming, the effect of air supply on reduction of the tar residual rate was more significant, while that on suppression of carbon particulate formation was smaller.     

Selective protein transport: Identity of the solubilized phosvitin receptor from chicken oocytes

By two independent methods, the solubilized receptor for phosvitin (PV) has a subunit MW of 116K. Affinity chromatography, showed that only 2 of the more than 25 proteins present in the total detergent solubilized oocyte membrane extract were retained on a PV–agarose column. These proteins of MW of 116K and 100K could be eluted from PV–agarose with free PV. By gel exclusion chromatography, the receptor-¹²⁵I-PV complexes elute in the void volume of a Biogel A-1.5 column. When these void fractions were assayed by SDS-PAGE only a single protein of MW of 116K was observed in addition to ¹²⁵I-PV.     

Effects of weak microwave fields amplitude modulated at ELF on EEG of symmetric brain areas in rats

Averaged electroencephalogram (EEG) frequency spectra were studied in eight unanesthetized and unmyorelaxed adult male rats with chronically implanted carbon electrodes in symmetrical somesthetic areas when a weak (0.1–0.2 mW/cm) microwave (MW, 945 MHz) field, amplitude-modulated at extremely low frequency (ELF) (4 Hz), was applied. Intermittent (1 min “On,” 1 min “Off”) field exposure (10-min duration) was used. Hemispheric asymmetry in frequency spectra (averaged data for 10 or 1 min) of an ongoing EEG was characterized by a power decrease in the 1.5–3 Hz range on the left hemisphere and by a power decrease in the 10–14 and 20–30 Hz ranges on the right hemisphere. No differences between control and exposure experiments were shown under these routines of data averaging. Significant elevations of EEG asymmetry in 10–14 Hz range were observed during the first 20 s after four from five onsets of the MW field, when averaged spectra were obtained for every 10 s. Under neither control nor pre- and postexposure conditions was this effect observed. These results are discussed with respect to interaction of MW fields with the EEG generators. Bioelectromagnetics 18:293–298, 1997. © 1997 Wiley-Liss, Inc.     

Fluidized-bed steam gasification of rice hull

Steam gasification of grain by-products can be a significant biomass conversion technology because of the need to utilize agricultural waste for non-food applications including energy resources. The most obvious beneficiary will be the developing countries whose economies are often tied to agricultural produce and are lacking in conventional fuels. One agricultural by-product that shows promise is the rice hull; it is found in abundance in the rice mills of producer countries and is considered as a waste material. Although gasification of rice hull has been proposed as a potential waste disposal and energy recovery method, little has been done to fully realize this proposition. In the present work, data were obtained for steam gasification of rice hull in a bench-scale fluidized-bed gasifier, a technology which has proven to be feasible for other grain by-products. The produced gas, which is rich in hydrogen, has been found to have a heating value ranging between 12.1 and 11.1 MJ m⁻³ at the respective reactor temperatures of 700 and 800°C; energy recovery varies between 35 and 59%.     

Complex Formation of Microbial Alkaline Protease Inhibitor (S-SI) with Subtilisin BPN′ and Its Properties

Microbial alkaline protease inhibitor, S–SI, was investigated on the interaction with subtilisin BPN′ Inhibitory equivalent of S–SI to subtilisin BPN′ was determined that one molecule of S–SI (MW = 23,000) inhibited two molecules of subtilisin BPN′ (MW = 27,700). The S–SI-subtilisin BPN′ complex was isolated by gel filtration on Sephadex G–100 and rhombic crystals were obtained. DIP- and ZAGPCK-subtilisin BPN′ did not form such complex with S–SI. Homogeneity of the complex was determined by disc electrophoresis. The isoelectric point of the complex was pH 5.5. Assay of S–SI dissociated and amino acid analysis of the complex indicated that one subunit (a half molecule) of S–SI was combined with one molecule of subtilisin BPN′ From molecular weight determination, it was clarified that the complex was composed of one molecule (consist of 2 subunits) of S–SI and two molecules of subtilisin BPN′.     

Molecular weight characterization of high molecular weight dextran with multiangle light scattering in on‐line and off‐line mode

This work reports the molecular weight (MW) analysis of high MW dextran using multiangle light scattering (MALS) in both chromatography and automated batch measurement mode. The results show that the chromatographic columns alter the high MW native dextran and cause underestimation of the MW as a consequence. Alternatively, a batch MALS measurement (without columns) provides more accurate MW values. The batch MALS measurement was automated with the incorporation of an automatic sample dilution and injection device. This automation reduces the sample preparation time and minimizes concentration errors introduced by manual sample dilution. To the best of our knowledge, this is the first study using an automated batch MALS in the analysis of high MW dextran. © 2015 Wiley Periodicals, Inc. Biopolymers 103: 387–392, 2015.     

Sequencing Regular and Labeled Oligonucleotides Using Enzymatic Digestion and Ionspray Mass Spectrometry

A method using a combination of enzymatic digestion and ionspray mass spectrometry (MS) was developed for sequencing oligodeoxynucleotides (ODNs) containing more than 20 bases. Phosphodiesterase (PDE) digestion of ODNs produced truncated ODNs whose molecular weights (MWs) were determined by ionspray MS. It was demonstrated that reconstruction of MW spectra over a large MW range produced easy-to-read sequence ladders similar to those obtained using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI–TOF-MS). Sample and enzyme cleanup, digestion control, and MW reconstruction were found to be crucial factors. For regular ODNs, both 5′- and 3′-PDE digestions are needed for complete sequencing. Late in the time course of PDE digestions, 5′-nucleoside monophosphates were found to produce artifactual peaks in the reconstructed MW spectra, and a table correlating base compositions and MS ions was compiled to handle such situations. For labeled ODNs, it is necessary to use collision-induced dissociation–tandem mass spectrometry (CID–MS/MS) for complete sequence determination. Sequencing of regular 22-mer and labeled 18-mer ODNs was demonstrated in this work.     

Soil temperatures during bushfires in semi-arid,mallee shrublands

Abstract Soil temperatures were measured during 11 experimental fires in semi-arid mallee shrublands in central NSW. Sensors were placed at depths from 1–10 cm beneath the soil surface in three fuel types; litter beneath Eucalyptus shrubs, live hummocks of the grass Triodia irritans and litter beneath shrubs of Acacia species. Weights of these fuels per unit area were determined. Maximum soil temperature and its duration were related to fuel type and depth. Mean weights of Eucalyptus and Triodia fuels were similar (0.35 kg m^?2), while there was less Acacia fuel (0.1 kg m^?2). Highest maximum temperatures were registered under Eucalyptus litter (e.g. 140°C at 2 cm). Maximum temperatures under Triodia and Acacia litter were similar (e.g. 60–70°C at 2 cm). Durations were examined in two temperature classes (60–120 and > 120°C) chosen to represent threshold for stimulation of germination and mortality, respectively, of soil-stored seeds. Temperatures between 60 and 120°C were recorded only between 0–2 cm soil depth for Acacia and Triodia (one exception at 4 cm). No temperatures >120°C were recorded for these fuel types. Temperatures between 60 and 120°C were recorded to 5 cm depth under Eucalyptus fuels while putative lethal temperatures for seeds occurred occasionally at 0–2 cm depth. The results indicated greatest potential for stimulation of germination and death of buried seeds under Eucalyptus fuels, although the level of variability of temperature was highest under Eucalyptus fuels. Despite similar fuel loads, differences between temperatures under Eucalyptus and Triodia fuels reflected the influence of the depth of the fuel bed, with Triodia hummocks constituting a deep fuel bed and Eucalyptus litter a shallow fuel bed.     

Molecular weight profiles of proanthocyanidin polymers

The MW profiles of proanthocyanidin polymers (condensed tannins) from 32 samples representing a wide range of plant tissues of many different species have been obtained by gel permeation chromatography of the peracetate derivatives. The tannins vary widely in MW, with M?_n values for the peracetates in the range 1600–5500. The MW profiles vary greatly from those with narrow, rather smooth distributions, to those which are discontinuous.     

Molecular Weight Determination of Gliadin Fractions in Gel Filtration by SDS-Polyacrylamide Gel Electrophoresis and Sedimentation Equilibrium

Gliadin was fractionated into three fractions; ω-gliadin, Fraction III (γ-gliadin) and Fraction IV (α- and β-gliadin). The determination of the molecular weights (MW) of the three fractions was performed by both SDS-polyacrylamide gel electrophoresis (SDS–PAGE) and sedimentation equilibrium. In SDS–PAGE, ω-gliadin gave three bands (MW 50,000, 54,000 and 64,000), Fraction III two bands (MW 38,000 and 46,000) and Fraction IV two bands (MW 33,000 and 38,000), The sedimentation analysis showed that each fraction was fairly homogeneous relative to molecular weight. The molecular weights obtained by sedimentation were 28,000 for Fraction III and 27,000 for both Fraction IV and ω-gliadin. The disagreement in molecular weight between sedimentation and gel electrophoresis was discussed.     

Mass and energy balance analyses of a downdraft gasifier

An empirical stoichiometric equation has been developed for wood chip gasification in a commercial-scale moving bed downdraft gasifier. The equation is based on an analysis of experimental overall and elemental material balance data obtained with the gasifier. A thermodynamic analysis of the gasifier has also been performed, based on the resultant empirical stoichiometric equation. The first-law and second-law thermodynamic efficiencies of the gasifier have been evaluated for four different operating modes at three different output temperatures.The resultant empirical stoichiometry is in agreement with the means of the experimental data within one standard deviation. The highest first-law and second-law thermodynamic efficiencies have been obtained when all products are considered useable; they are 90% and 62%, respectively. The lowest first-law and second-law efficiencies have been obtained when cool dry gas is considered as the only useable product; they are 72% and 53%, respectively. The heat loss from the system to the surroundings has been estimated to be 10% of the energy input; this corresponds to a loss of 7% of the available energy input. The available energy dissipation in the system, due to the various irreversibilities of the gasification process, has been evaluated to be 31% of the available energy input. This dissipation is not recoverable and reflects the nature of the process.     

Simulated sea level change alters anatomy, physiology, growth, and reproduction of red mangrove (Rhizophora mangle L.)

Tropical coastal forests – mangroves – will be one of the first ecosystems to be affected by altered sea levels accompanying global climate change. Responses of mangrove forests to changing sea levels depend on reactions of individual plants, yet such responses have not been addressed experimentally. We report data from a long-term greenhouse study that assessed physiological and individual growth responses of the dominant neotropical mangrove, Rhizophora mangle, to levels of inundation expected to occur in the Caribbean within 50–100 years. In this study, we grew potted plants in tanks with simulated semidiurnal (twice daily) high tides that approximated current conditions (MW plants), a 16-cm increase in sea level (LW plants), and a 16-cm decrease in sea level (HW plants). The experiment lasted 2½ years, beginning with mangrove seedlings and terminating after plants began to reproduce. Environmental (air temperature, relative humidity, photosynthetically active radiation) and edaphic conditions (pH, redox, soil sulfide) approximated field conditions in Belize, the source locale for the seedlings. HW plants were shorter and narrower, and produced fewer branches and leaves, responses correlated with the development of acid-sulfide soils in their pots. LW plants initially grew more rapidly than MW plants. However, the growth of LW plants slowed dramatically once they reached the sapling stage, and by the end of the experiment, MW plants were 10–20% larger in all measured growth parameters. Plants did not exhibit differences in allometric growth as a function of inundation. Anatomical characteristics of leaves did not differ among treatments. Both foliar C:N and root porosity decreased from LW through MW to HW. Relative to LW and HW plants, MW plants had 1–7% fewer stomata/mm², 6–21% greater maximum photosynthetic rates, 3–23% greater absolute relative growth rates (RGRs), and a 30% higher RGR for a given increase in net assimilation rate. Reduced growth of R. mangle under realistic conditions approximating future inundation depths likely will temper projected increased growth of this species under concomitant increases in the atmospheric concentration of CO₂.     

Detection of Microtubules of the Flagellate Trichomonas vaginalis by Monoclonal Antibodies Specific for β-Tubulin1

ABSTRACT. Monoclonal antibodies specific for mammalian β-tubulin recognized the microtubule cytoskeleton of the flagellated protozoon Trichomonas vaginalis. Of seven antibodies, two demonstrated the axostyle, costa, recurrent flagellum, and anterior flagella by indirect immunofluorescence microscopy. The remaining five stained a hazy reticular pattern in the cytoplasm of formaldehyde-fixed, detergent-extracted organisms. Western immunoblots of whole T. vaginalis extracts treated with protease inhibitors and electrophoresed on polyacrylamide gels containing sodium dodecyl sulfate showed a major band at molecular weight 50,000 when probed with only one of the antibodies which stained the axial cytoskeleton. The antibodies which stained only the cytoplasm showed a different western blot pattern with a major doublet band at MW 58,000–60,000. Another antibody, which stained both the axial cytoskeleton and the reticular cytoplasmic pattern showed major bands at MW 58,000–60,000 and also at MW 40,000–42,000. The recognition of microtubule populations in T. vaginalis by these monoclonal antibodies was different than we found earlier with Leishmania donovani and Toxoplasma gondii, where all seven antibodies recognize cytoskeletal microtubules and produce western blots characteristic of tubulin. Only one of these seven antibodies recognizes tubulin in T. vaginalis by immunoblot. The microtubules of T. vaginalis do not demonstrate all epitopes recognized by monoclonal antibodies specific for mammalian β-tubulin; one of the antibodies appears to recognize an epitope which is morphologically associated with microtubules but does not have the characteristic MW of tubulin.     

Catalytic activity and the stability of horseradish peroxidase increase as a result of its incorporation into a polyelectrolyte complex with chitosan

The incorporation of horseradish peroxidase into polyelectrolyte complexes with chitosans of different molecular weights (MW 5–150 kDa) yielded highly active and stable enzyme preparations. As a result of the selection of optimal conditions for the formation of peroxidase-chitosan complexes, it was found that 0.1% chitosan with a MW of 10 kDa had the strongest activatory effect on peroxidase (activation degree, >70%) in the reaction of o-dianisidine oxidation by hydrogen peroxide. The complex formed by 0.001% chitosan with a molecular weight of 150 kDa was most stable: when immobilized on foamed polyurethane, it retained at least 50% of the initial activity for 550 days. The highest catalytic activity was exhibited in a 0.05 M phthalate buffer (pH 5.9–6.2) by the complex containing 0.006–0.009% chitosan in the indicator reaction. The activatory effect of the polysaccharide on the enzyme was determined by its influence on the binding and conversion of the reducting substrate peroxidase.     

Preparation of Long-Acting Superoxide Dismutase Using High Molecular Weight Polyethylene Glycol (41,000-72,000 Daltons)

Superoxide dismutase (SOD) has demonstrated therapeutic potential for treating a variety of conditions including radiation injury, oxygen toxicity, reperfusion injury, and inflammation, especially arthritis. However, the native enzyme's short half-life in plasma (6 minutes in mice. 25 minutes in man) limits the enzyme's effectiveness in many applications, or requires infusion of large doses. High doses of SOD derived from either natural or rDNA sources may increase the potential for immunologic sensitization. One effective use of native SOD is intra particular administration for treatment of arthritis, where injection of SOD into joints retards elimination (15 hour terminal half-life), allowing the effective use of lower doses.

To overcome the limitations resulting from rapid clearance. various researchers have increased the persistence of SOD by cross-linking SOD or by attaching polymeric substances, including dextrans, albumin, Ficoll, polyvinyl alcohol or polyethylene glycol (PEG). PEG is relatively safe; however. the amount of modification by PEG. is the MW range 1.900–5.000 daltons, which is necessary to optimally increase serum persistence and reduce immunogenicity, results in the loss of much of the enzymatic activity.

In this report we describe the preparation of SOD adducts containing I to 4 strands of high MW PEG (41,000–72,000 daltons). The MW range of these adducts, measured by steric exclusion HPLC based on protein standards, is 200,000 to over 1,100,000 daltons. The number of PEG strands attached per SOD dimer (32,000 daltons) was measured by HPLC. Because of the low degree of protein modification required to produce very high MW products, these PEG-SODS retain 90%-100% of the SOD activity of the native enzyme. Additionally, these very large adducts demonstrate longer persistence and lower immunogenicity and antigenicity compared to the more highly modified PEG-SODS containing low MW PEG (i.e., 7–16 strands of 5.000 dalton methoxy-PEG).