Determine effect of pressure heating on carbohydrate related molecular structures in association with carbohydrate metabolic profiles of cool-climate chickpeas using Globar spectroscopy.

Grain has been heat-processed to alter rumen degradation characteristics and improve nutrient availabilities for ruminants. However, limited study was found on internal structure changes induced by processing on a molecular basis. The objectives of this study were to use advanced vibrational molecular spectroscopy to: (1) determine the processing induced carbohydrate (CHO) structure changes on a molecular basis, (2) investigate the effect of pressure heating on changes of CHO chemical profiles, CHO subfractions in cool-climate CDC Chickpea varieties, and (3) to reveal the association between carbohydrates related molecular spectra with carbohydrate metabolic profiles. The cool-climate CDC chickpea varieties with multisource were pressure heated in an autoclave at 120 °C for 60 min; and FTIR vibrational spectroscopy was used to detect the molecular spectra. Molecular spectroscopic results showed that compared to raw chickpea varieties, autoclave heating induced changes in both total CHO (region and baseline ca. 1186-946 cm-1) and structural CHO (STCHO, region and baseline ca. 1482-1186 cm-1), except for cellulosic compounds (CELC, region and baseline ca. 1374-1212 cm-1). The CHO chemical profile and rumen degradation results showed that autoclave heating decreased rumen degradable, undegradable and intestinal digestible sugar (CA4) content, but increased available fiber (CB3) content, without affecting available energy of chickpeas. The changes of CHO molecular spectra in chickpea varieties were strongly correlated with CHO chemical profiles, CHO subfractions, and CHO rumen degradation characteristics. Moreover, the regression analysis showed that STCHO peak 1 height could be used to predict sugar content, its rumen degradability and digestibility of chickpeas. Our results suggest that autoclave heating markedly changes sugar and fiber degradation characteristics. The carbohydrate molecular spectral profiles are associated with carbohydrate metabolic profiles in raw and pressure heated cool-climate chickpeas.

Effective atomic number of some sugars and amino acids for scattering of (241)Am and (137)Cs gamma rays at low momentum transfer.

In this paper, we report the effective atomic number of some H, C, N and O based sugars and amino acids. These have been determined by using a handy expression which is based on the theoretical angle integrated small angle (coherent+incoherent) scattering cross sections of seven elements of Z≤13 in four angular ranges of (0-4°), (0-6°), (0-8°) and (0-10°)for (241)Am (59.54 keV) and (137)Cs (661.6 keV) gamma rays. The theoretical scattering cross sections were computed by a suitable numerical integration of the atomic form factor and incoherent scattering function compilations of Hubbell et al. (1975) which make use of the non-relativistic Hartree-Fock (NRHF) model for the atomic charge distribution of the elements in the angular ranges of interest. The angle integrated small angle scattering cross sections of the H, C, N and O based sugars and amino acids measured by a new method reported recently by the authors were used in the handy expression to derive their effective atomic number. The results are compared with the other available data and discussed. Possible conclusions are drawn based on the present study.

Investigation of photochemical reactions of saccharides during direct ultraviolet absorbance detection in capillary electrophoresis.

Direct UV detection of saccharides in capillary electrophoresis is possible due to a base-catalyzed conversion into UV absorbing substances initiated by the light of the UV detector lamp. In the present study the compounds formed during this reaction were investigated with capillary electrophoresis using an additional UV lamp for online irradiation at a certain distance before the detector resulting in a separation of the conversion products. It was found that for all investigated saccharides (glucose, ribose and sucrose) the major portion of the UV absorption in direct UV detection resulted from one and the same substance. By CE-UV, CE-MS as well as HPLC-MS measurements this substance was demonstrated to be malondialdehyde, present as its conjugate base malonenolate under alkaline conditions. Additional experiments revealed that the conversion reaction is highly sensitive to the residence time under the UV light as malondialdehyde can further degrade into non-UV absorbing products. NMR measurements showed that under alkaline conditions this degradation proceeds slowly even without UV irradiation.

Carbohydrate radicals: from ethylene glycol to DNA strand breakage.

Radiation-induced DNA strand breakage results from the reactions of radicals formed at the sugar moiety of DNA. In order to elucidate the mechanism of this reaction investigations were first performed on low molecular weight model systems. Results from studies on deoxygenated aqueous solutions of ethylene glycol, 2-deoxy-d-ribose and other carbohydrates and, more relevantly, of d-ribose-5-phosphate have shown that substituents can be eliminated from the ß-position of the radical site either proton and base-assisted (as in the case of the OH substituent), or spontaneously (as in the case of the phosphate substituent). In DNA the C(4') radical undergoes strand breakage via this type of reaction. In the presence of oxygen the carbon-centred radicals are rapidly converted into the corresponding peroxyl radicals. Again, low molecular weights models have been investigated to elucidate the key reactions. A typical reaction of DNA peroxyl radicals is the fragmentation of the C(4')-C(S') bond, a reaction not observed in the absence of oxygen. Although OH radicals may be the important direct precursors of the sugar radicals of DNA, results obtained with poly(U) indicate that base radicals may well be of even greater importance. The base radicals, formed by addition of the water radicals (H and OH) to the bases would in their turn attack the sugar moiety to produce sugar radicals which then give rise to strand breakage and base release. For a better understanding of strand break formation it is therefore necessary to investigate in more detail the reactions of the base radicals. For a start, the radiolysis of uracil in oxygenated solutions has been reinvestigated, and it has been shown that the major peroxyl radical in this system undergoes base-catalysed elimination of [Formula: see text], a reaction that involves the proton at N(l). In the nucleic acids the pyrimidines are bound at N(l) to the sugar moiety and this type of reaction can no longer occur. Therefore, with respect to the nucleic acids, pyrimidines are good models only in acid solutions where the [Formula: see text] elimination reaction is too slow to compete with the bimolecular reactions of the peroxyl radicals. Moreover, the long lifetime of the radical sites on the nucleic acid strand may allow reactions to occur which are kinetically of first order, and which cannot be studied in model systems at ordinary dose rates. It is therefore suggested to extend model system studies to low dose rates and to oligonucleo-tides. Such studies might eventually reveal the key reactions in radical-induced DNA degradation.

Reduced contribution of thermally labile sugar lesions to DNA double strand break formation after exposure to heavy ions.

In cells exposed to low linear energy transfer (LET) ionizing-radiation (IR), double-strand-breaks (DSBs) form within clustered-damage-sites (CDSs) from lesions disrupting the DNA sugar-phosphate backbone. It is commonly assumed that all DSBs form promptly and are immediately detected by the cellular DNA-damage-response (DDR) apparatus. However, there is evidence that the pool of DSBs detected by physical methods, such as pulsed-field gel electrophoresis (PFGE), comprises not only promptly forming DSBs (prDSBs) but also DSBs developing during lysis at high temperatures from thermally-labile sugar-lesions (TLSLs). We recently demonstrated that conversion of TLSLs to DNA breaks and ultimately to DSBs also occurs in cells during the first hour of post-irradiation incubation at physiological temperatures. Thus, TLSL-dependent DSBs (tlDSBs) are not an avoidable technique-related artifact, but a reality the cell always faces. The biological consequences of tlDSBs and the dependence of their formation on LET require in-depth investigation. Heavy-ions (HI) are a promising high-LET radiation modality used in cancer treatment. HI are also encountered in space and generate serious radiation protection problems to prolonged space missions. Here, we study, therefore, the effect of HI on the yields of tlDSBs and prDSBs. We report a reduction in the yield of tlDBSs stronger than that earlier reported for neutrons, and with pronounced cell line dependence. We conclude that with increasing LET the complexity of CDSs increases resulting in a commensurate increase in the yield prDSBs and a decrease in tlDSBs. The consequences of these effects to the relative biological effectiveness are discussed.

Ultrasound assisted extraction of carbohydrates from microalgae as feedstock for yeast fermentation.

Recently, carbohydrates biomass from microalgae is considered as a promising and inexpensive feedstock for biofeuls production by microorganism fermentation. The main obstacle of the process is microalgae pretreatment and carbohydrates extraction from algal cell. In this study, comparison of three pretreatment methods was performed and the results showed that ultrasonic assisted extraction (UAE) was very effective. The effects of four parameters (ultrasonic power, extraction time, flow rate and algal cell concentration, respectively) on extraction efficiency were also investigated. Additionally, in order to identify significant factors for glucose yield, combination of these four parameters was examined by using fractional factorial design (FFD) and the regression model was obtained. Meanwhile, the refined model was confirmed as a good fitting model via analysis of variance (ANOVA). After extraction, glucose obtained from microalgae was used as substrate for Rhodosporidium toruloides fermentation and yeast biomass was much higher than that of control culture.

Microwave modification of sugar cane to enhance juice extraction during milling.

Sugar extraction from cane requires shredding and crushing, both of which are energy intensive activities. Cane shredders account for almost 30% of the total power requirements for the juice extraction train in a sugar mill with four mills. Shredder hammers also wear quickly during the crushing season and need to be regularly maintained or replaced. Microwave pre-treatment of other plant based materials has resulted in significant reductions in total processing energy. This paper briefly reviews the underlying structure of sugar cane and how microwave pre-treatment may interact with sugar cane. Microwave treatment reduced the strength of sugar cane samples to 20% of its untreated value. This strength reduction makes it easier to crush the cane and leads to a 320% increase in juice yield compared with untreated cane when cane samples were crushed in a press. There was also a 68% increase in Brix %, a 58% increase in total dissolved solids, a 58% reduction in diffusion time, a 39% increase in Pol%, and a 7% increase in juice purity compared with the control samples after 60 minutes of diffusion in distilled water.

Ultrasonic pretreatment of corn slurry for saccharification: a comparison of batch and continuous systems.

The effects of ultrasound on corn slurry saccharification yield and particle size distribution was studied in both batch and continuous-flow ultrasonic systems operating at a frequency of 20 kHz. Ground corn slurry (28%w/v) was prepared and sonicated in batches at various amplitudes (192-320 microm(peak-to-peak (p-p))) for 20 or 40s using a catenoidal horn. Continuous flow experiments were conducted by pumping corn slurry at various flow rates (10-28 l/min) through an ultrasonic reactor at constant amplitude of 12 microm(p-p). The reactor was equipped with a donut shaped horn. After ultrasonic treatment, commercial alpha- and gluco-amylases (STARGEN 001) were added to the samples, and liquefaction and saccharification proceeded for 3h. The sonicated samples were found to yield 2-3 times more reducing sugars than unsonicated controls. Although the continuous flow treatments released less reducing sugar compared to the batch systems, the continuous flow process was more energy efficient. The reduction of particle size due to sonication was approximately proportional to the dissipated ultrasonic energy regardless of the type of system used. Scanning electron microscopy (SEM) images were also used to observe the disruption of corn particles after sonication. Overall, the study suggests that both batch and continuous ultrasonication enhanced saccharification yields and reduced the particle size of corn slurry. However, due to the large volume involve in full scale processes, an ultrasonic continuous system is recommended.

Photoimmobilization of unmodified carbohydrates on activated surface.

Herein, we demonstrate a simple, versatile and efficient method for immobilization of unmodified carbohydrates onto a solid surface. The method employs a photoreactive-cellulose membrane which is prepared by the introduction of a photoreactive group to a cellulose membrane through 1-fluoro-2-nitro-4-azidobenzene (FNAB). Upon exposure to UV light photo-reactive azido group of the activated cellulose membrane transforms itself into highly reactive nitrene which covalently binds to underivatized carbohydrates through an insertion reaction. Maximum immobilization of carbohydrate is achieved at the UV exposure time of 60min and carbohydrate concentration of 100microg/disk. The immobilized carbohydrate is assayed by affinity binding of biotin-labeled lectins. The amount of Con A required for detecting immobilized carbohydrate ranges from 4-10microg/ml. The binding affinities of the lectin to the immobilized carbohydrates are analyzed by calculating their IC(50) values. Overall, the present work demonstrates an efficient immobilization of underivatized carbohydrate onto cellulose surface and has the potential to be applied to other surfaces.

Studies on effective atomic number, electron density and kerma for some fatty acids and carbohydrates.

The effective atomic number, Z(eff), the effective electron density, N(el), and kerma have been calculated for some fatty acids and carbohydrates for photon interaction in the extended energy range from 1 keV to 100 GeV using an accurate database of photon-interaction cross sections and the WinXCom program. The significant variation of Z(eff) and N(el) is due to the variations in the dominance of different interaction processes in different energy regions. The maximum values of Z(eff) and N(el) are found in the low-energy range, where photoelectric absorption is the main interaction process. The minimum values of Z(eff) and N(el) are found at intermediate energies, typically 0.05 MeV < E < 5 MeV, where Compton scattering is dominant. In this case, Z(eff) is equal to the mean atomic number of the bio-molecule. Wherever possible, the calculations are compared with experimental results. A comparison is also made with the single values of the Z(eff) and N(el) provided by the program XMuDat. It is also observed that carbohydrates have a larger kerma than fatty acids in the low-energy region, where photoelectric absorption dominates. In contrast, fatty acids have a larger kerma than carbohydrates in the MeV range, where Compton scattering is the main interaction process.

Indium triflate catalyzed peracetylation of carbohydrates.

Peracetylation is a very common protection strategy that is widely implemented in carbohydrate synthesis. Here, a method for the peracetylation of carbohydrates using catalytic In(OTf)(3) in neat acetic anhydride is reported. In(OTf)(3) has low toxicity and is mild and water tolerant, and the reactions are high yielding and efficient. Details regarding the scope and mechanism of the reaction are briefly discussed.

High-energy glycoconjugates: synthetic transformations of carbohydrates using microwave and ultrasonic energy.

Methods to transform carbohydrates are often complex and tedious, both due to the vast array of naturally occurring and synthetically designed scaffolds which may manifest meager to drastic reactivity, dependent upon the transformation sought and the stereogenic site chosen. In order to facilitate and expedite desired synthetic transformation, many researchers are utililizing microwave and ultrasonic irradiation to achieve their goals, in generally high yields within a shorter period of time, and often without undesirous byproducts. The basic physical principles underlying the energy regimes are qualitatively discussed prior to review of the applications in carbohydrate syntheses and transformation. This literature review looks at research involving glycosylations, -OH group conversions, isotopic incorporation, and C-N bond formation. Instances of improved yields and selectivities resultant from the use of these high-energy sources will be highlighted.

Formation of sugar radicals in RNA model systems and oligomers via excitation of guanine cation radical.

In previous work, we have shown that photoexcitation of guanine cation radical (G*+) in frozen aqueous solutions of DNA and its model compounds at 143 K results in the formation of neutral sugar radicals with substantial yield. In this report, we present electron spin resonance (ESR) and theoretical (DFT) evidence regarding the formation of sugar radicals after photoexcitation of guanine cation radical (G*+) in frozen aqueous solutions of one-electron-oxidized RNA model compounds (nucleosides, nucleotides and oligomers) at 143 K. Specific sugar radicals C5'*, C3'* and C1'* were identified employing derivatives of Guo deuterated at specific sites in the sugar moiety, namely, C1'-, C2'-, C3'- and C5'-. These results suggest C2'* is not formed upon photoexcitation of G*+ in one-electron-oxidized Guo and deuterated Guo derivatives. Phosphate substitution at C5'- (i.e., in 5-GMP) hinders formation of C5'* via photoexcitation at 143 K but not at 77 K. For the RNA-oligomers studied, we observe on photoexcitation of oligomer-G*+ the formation of mainly C1'* and an unidentified radical with a ca. 28 G doublet. The hyperfine coupling constants of each of the possible sugar radicals were calculated employing the DFT B3LYP/6-31G* approach for comparison to experiment. This work shows that formation of specific neutral sugar radicals occurs via photoexcitation of guanine cation radical (G*+) in RNA systems but not by photoexcitation of its N1 deprotonated species (G(-H)*). Thus, our mechanism regarding neutral sugar formation via photoexcitation of base cation radicals in DNA appears to be valid for RNA systems as well.

Microwave energy: a versatile tool for the biosciences.

As the range of techniques for microwave heating has expanded, so have the areas in which it can have a profound impact. Two emerging areas are the application of microwave heating for the synthesis of peptides, peptoids, oligopeptides and carbohydrates and in the field of proteomics.

Sonochemistry: a powerful way of enhancing the efficiency of carbohydrate synthesis.

Using sonication as a means of facilitating organic reactions in carbohydrate chemistry was explored under the conditions used for traditional organic synthesis. An array of representative reactions, including hydroxy group manipulation (acylation, protection/deprotection, acyl group migration), thioglycoside synthesis, azidoglycoside synthesis, 1,3-dipolar cycloaddition and reductive cleavage of benzylidene, commonly used in the synthesis of carbohydrate derivatives was examined. A series of glycosylation reactions that employ thioglycosides, glycosyl trichloroacetimidate, glycosyl bromide and glycosyl acetate as the glycosyl donors was also examined. Our results demonstrate that sonication can significantly shorten the reaction time, enhance the reactivity of reactant and lead to superior yield and excellent stereoselectivity. More importantly, a general protocol of glycosylation may finally be developed. Sonication is compatible to the conditions used for traditional organic synthesis. We believe that sonication can also be applied to other areas of synthetic processes.

Study of materials for mixed field dosimetry by EPR spectroscopy.

EPR dose reconstruction after accidental photon exposure based on materials irradiated in the vicinity of the victim (sucrose, medicine tablets, etc.) was used successfully in several cases referenced in the literature. However, accidental exposure may also occur with a neutron component such as in the Tokai-Mura criticality accident. The aim of this work is to investigate the potentiality of EPR dosimetry for mixed photon and neutron field exposure with different organic materials already used for photon exposure (sucrose) or with potential dosimetric properties (ascorbic acid, sorbitol, glucose, galactose, fructose, lactose and mannose). To assess the neutron sensitivity, the materials were exposed to a mixed radiation field of an experimental reactor with different neutron to photon ratios. The relative neutron sensitivity was found to range from 12 to 43% according to the materials. The potentiality of these materials for mixed field EPR dosimetry is discussed.

Photo-induced hole transfer from base to sugar in DNA: relationship to primary radiation damage.

This work presents the hypothesis that photo-excitation of G.+ in DNA and model systems results in the same electronic states expected from direct ionization of the sugar phosphate backbone and that these states lead to specific sugar radicals on the DNA sugar phosphate backbone. As evidence we show that visible photo-excitation of guanine cation radicals (G.+) in the dinucleoside phosphate TpdG results in high yields (about 85%) of deoxyribose sugar radicals at the C1' and C3' sites. Further, we have calculated transition energies of hole transfer from G.+ in TpdG using time-dependent density functional theory (TD-DFT) at the B3LYP/6-31G(d) level in gas phase as well as in a solvated environment. These calculations clearly predict that visible excitation of G.+ in TpdG causes transitions from only inner-shell filled molecular orbitals (MOs) to the singly occupied molecular orbital (SOMO) that effectively result in hole transfer from guanine either to the sugar phosphate backbone or to the adjacent base, thymine. The hole transfer is followed by rapid deprotonation from the sugar to form C1' and C3' radicals. These experimental and theoretical results are in agreement with our previously published experimental and theoretical results that photo-excitation of G.+ results in high yields of deoxyribose sugar radicals in DNA, guanine deoxyribonucleosides and deoxyribonucleotides. Photo-excitation of G.+ therefore provides a convenient method to produce and study sugar radicals that are expected to be formed in gamma-irradiated DNA systems unencumbered by the many other pathways involved in direct ionization.

High-light-induced changes on photosynthesis, pigments, sugars, lipids and antioxidant enzymes in freshwater (Nostoc spongiaeforme) and marine (Phormidium corium) cyanobacteria.

We studied the effects of high-light exposure (500 micromol m(-2) s(-1) of photosynthetic active radiation) on the cyanobacteria Nostoc spongiaeforme Agardh, a fresh-water alga, and Phormidium corium Agardh (Gomont), a marine alga, with respect to photosynthesis, pigments, sugar content, lipid peroxidation, fatty acids composition, antioxidant enzymes activity and DNA. It was seen that the ratio of variable fluorescence (Fv) to maximum fluorescence (Fm), which is indicative of photosynthetic efficiency, decreased because of the light treatment. The damage to photosynthesis occurred in the antenna system and the photosynthetic II reaction center. Photobleaching of photosynthetic pigments was also observed. High-light treatment also resulted in decreased sugar content, which was probably due to the effect on photosynthesis. Peroxidation of membrane lipids, indicating oxidative damage to lipids and a high level of unsaturation in the cell membrane, was also observed. The activity of antioxidant enzyme superoxide dismutase and ascorbate peroxidase was increased, probably as a result of oxidative damage observed in the form of lipid peroxidation. Quantitative decreases in phospholipid and glycolipid levels were also observed. The level of unsaturated fatty acids in total lipids and glycolipids remained unchanged in both species; however, the level of saturated fatty acids decreased, which slightly changed the ratio in favor of unsaturated fatty acids. Degradation of DNA was also observed in both species. There was a transient plateau 2-4 h after exposure to high-light treatment in the Fv/Fm ratio and in levels of phycobilisome pigments, sugars and antioxidant enzymes after an initial decrease 1 h after the treatment. These findings may indicate a period of partial adaptation to high light that is due to the efficiency of protective processes operational in the two species, which subsequently failed after a longer exposure duration of 4-6 h.

Homolytic cleavage of the O-glycoside bond in carbohydrates: a steady-state radiolysis study.

The formation of products resulting from the O-glycoside bond cleavage following radiolysis of aqueous solutions of methyl-alpha-D-glucopyranoside (I), 3-O-methyl-alpha-D-glucopyranose (II), maltose, lactose, gentiobiose and cellobiose were studied. Radiation-induced destruction yields were also determined for dextran, laminarin and trimethylcelulose upon irradiation of their aqueous solutions. Oxygen, quinones and compounds capable of forming quinoid structures were found to inhibit radiation-induced homolytic destruction processes taking place in glycosides, di- and polysaccharides. The data obtained in this study enabled the authors to demonstrate an important role played by the fragmentation reaction of C-2 radicals which were generated from the starting substances in the formation of final radiolysis products.