Recent Advances in the Development of a Diagnostic Test for Irradiated Foodstuffs

Recent advances in the development and application of diagnostic tests for irradiated foodstuffs are reviewed. Exposure of water, the major chemical constituent of most foodstuffs to a source of ionising radiation initially generates the highly reactive radical species H', -OH and e^- (aq) which react very rapidly with a wide variety of biological molecules. The detection of foodstuffs subjected to irradiation processing requires the identification and/or quantification of ‘unnatural’ chemical species (i.e. those not usually formed by normal metabolic processes) produced by the attack of ·OH radical or e^-(aq) on suitable ‘target’ molecules. Modern methods for the analysis of a series of these ‘unnatural’ products arising from the interaction of radiolytically-generated ·OH radical or e^-(aq) with polyunsaturated fatty acids, DNA, aromatic compounds and other biologically important scavenger molecules are examined. It is concluded that the analytical test to be conducted is highly dependent on the nature of the foodstuff to be tested.     

Imaging of subcellular structures by scanning ion microscopy and mass spectrometry. Advantage of cryofixation and freeze substitution procedure over chemical preparation

With the IMS 4F, a scanning ion microscope and mass spectrometer (SIMS), it is possible to map chemical elements with a lateral resolution of about 250 nm over a field of view of 50 × 50 μm². Such conditions should enable the imaging of subcellular structures with constitutive ionic species such as CN^?, P^?, S^?. The study was performed on heart and renal tissues prepared either by chemical procedure or cryofixation-freeze substitution (CF-FS) prior to embedding. Heart tissue was chosen because cardiocytes display a simple structural organization whereas the structural organization of kidney tubular cells is more complex. Whatever the preparation procedure, nuclei were easily identified due to their high P^? content. The CN^?, P^?, and S^? ion images obtained on heart and renal tissues prepared by chemical procedure showed weak contrasts inside the cytoplasm so that it was difficult to recognize the organelles. After CF-FS, enhanced contrasted images allow organelle (mitochondria, myofibrils, lysosomes, vacuoles, basal lamina, etc) characterization. This work demonstrated that CF-FS is a more suitable preparation procedure than chemical method to reveal organelle structures by their chemical composition. The improvements in the imaging of these structures is an essential step to establish the correlation between the localization of a trace element (or a molecule tagged with isotopes or particular atoms) and its subcellular targets.     

Soil nitrogen dynamics and plant-induced soil changes under plantations and primary forest in lowland Amazonia, Brazil

The influence of plant species on soil nitrogen (N) dynamics was investigated in lowland Amazonia, Brazil under plantations of tree species with varied phenologies, resource requirements, and chemical characteristics in fine litter. Seasonal N dynamics were studied in replicated stands of Pinus caribaea var. hondurensis Barrett & Golfari, Euxylophora paraensis Hub., Carapa guianensis Aubl., a Leguminosae combination (Dalbergia nigra Fr. All., Dinizia excelsa Ducke, Parkia multijuga Benth.), and native forest in the Curuá-Una Forest Reserve, Pará, Brazil. Textural, mineralogical, and chemical soil properties at 1 m depth under the plantations and the forest indicated that initial soil properties were similar. Net annual N mineralization ranged from 195 kg ha^-1 (P. caribaea) to 328 kg ha^-1 (forest), and was related to fine root N contents in the surface root mat (R² = 0.96, p = 0.01). Net annual N mineralization was also inversely related to within-stand nitrogen-use efficiency (R² = 0.81, p = 0.04). These results suggest that tree species or groups of species with varied N-use efficiencies altered soil N transformation rates in a predictable manner.     

Fine Root Morphology,Biochemistry and Litter Quality Indices of Fast- and Slow-growing Woody Species in Ethiopian Highland Forest

Fine root turnover of trees is a major C input to soil. However, the quality of litter input is influenced by root morphological traits and tissue chemical composition. In this study, fine roots of ten tropical woody species were collected from an Afromontane forest in the northern highlands of Ethiopia. The fine roots were analysed for root morphological traits and tissue chemistry measured as proxy carbon fractionations. Based on stem increment, the 10 species were divided into faster- and slower-growing species. Faster-growing species exhibited higher specific root length (1362 cm g^?1) than slower-growing species (923 cm g^?1). Similarly specific root area was higher in faster-growing species (223 cm² g^?1) than in slower-growing species (167 cm² g^?1). Among the carbon fractions, the acid-insoluble fraction (AIF) was the highest (44–51%). The carbon content, AIF, and the lignocellulose index were higher for slower-growing species. Root tissue density was lower in faster-growing species (0.33 g cm^?3) than slower-growing species (0.40 g cm^?3) and showed a strong positive correlation with carbon content (r ² = 0.84) and the AIF (r _pearson = 0.93). The morphological traits of fine roots between faster- and slower-growing species reflect the ecological strategy they employ. Slower-growing species have a higher tissue density which may reflect a greater longevity.     

Essential Oils from Piper caldense C. DC. and Piper xylosteoides (Kunth) Steud.: Seasonal Variation of the Chemical Composition and Antimicrobial Activity

The essential oils (EOs) chemical composition can be affected by several environmental factors, impacting their desired biological activities. In this sense, this work aimed to evaluate the seasonal variation of the chemical composition and antimicrobial activity of Piper caldense and Piper xylosteoides leaves EOs. Their chemical composition was determined by GC/MS and GC-FID analyses, resulting in the identification of eighty compounds. P. caldense EOs were mainly consisted of sesquiterpene hydrocarbons, whereas in P. xylosteoides EOs, monoterpene hydrocarbons were predominant. EOs from both species strongly inhibited B. subtilis (MIC=0.25 mg mL⁻¹), while only P. caldense EOs showed strong activity against S. aureus (MIC=0.50 mg mL⁻¹). P. caldense spring EO showed the broadest spectrum of antimicrobial action amongst all samples. For each species, PCA seasonally differentiated EOs chemical composition. In addition, as expected, PCA of all samples showed a distinction between the two species. This study has successfully demonstrated the importance of evaluating the seasonal variation of EOs chemical composition and antimicrobial activity in obtaining a product with the desired properties.     

Dynamics of trace metals in the system water – soil – plant – wild rats – tapeworms (Hymenolepis diminuta) in Maglizh area,Bulgaria

BackgroundThe impact of chemical elements on the biosphere is a function of their concentration and chemical form. Elucidation and prognosing of the latters in water basins and soil extracts is of particular significance for the assessment of their bioaccumulation in plants and animals.ObjectivesTrace metals dynamics in the system water – soil–plant–wild rats – Hymenolepis diminuta in two agro-industrial zones (East and West) around Maglizh city, Bulgaria were investigated through experimental studies and thermodynamic modelling of the chemical species.MethodsSamples from surface waters of rivers, their nearby uncultivated soils, meadow uncultivated vegetation (Ranunculus acris and Gramineae) and field rats were collected. In situ measurements and laboratory analyses were performed for the determination of the physico-chemical characteristics and total concentrations of Al, Fe, Mn, Ni, Cu, Zn and Pb. The distribution of their dissolved chemical species in water samples and in the aqueous soil extracts was calculated using a thermodynamic approach. The relationship chemical species - bioaccumulation was discussed.ResultsWaters and soils in the East zone of Maglizh area were found to be more polluted compared to those in the West one, regarding Ni, Mn, Zn, Pb and Cu, while Mn and Cu displayed the highest mobility in West zone soils. Trace metals contents in Ranunculus acris exceed that in Gramineae, since the highest accumulation factors were calculated for Cu and Zn. The highest accumulation in rats was found for Zn followed by Cu, being higher in the West zone. Thermodynamic modelling shows that Mn²⁺ free ions are dominant in both waters and aqueous soil extracts. Ni²⁺ and Zn²⁺ ions followed by metal-organic complexes are dominant in waters of East zone while metal-organic complexes followed by free ions are dominant in waters of West zone and both soil extracts. Metal-organic complexes are dominant for Fe, Cu and Pb in all samples studied, while mainly hydroxy forms (Al(OH)₄⁻) followed by metal-organic complexes are typically for Al depending on pH.ConclusionsExperimentally established bioaccumulation of trace metals in the studied vegetation and rats is a consequence of the total concentration of trace metals in waters and soils, their mobility and chemical species. The dominance of organic complexes of trace metals is a prerequisite for their bioaccumulation in plants. Rats are in direct contact with the soil solution and therefore, of importance is the content of free ions of Mn²⁺, Ni²⁺, Zn²⁺, which are easily absorbed through the skin. The host-helminth system wild rat/H. diminuta could be used as a bioindicator for trace metals pollution.     

Chemical composition and construction cost for roots of Mediterranean trees, shrub species and grassland communities

The construction cost of fine roots was studied in 23 woody species and two grassland communities, growing under natural conditions in southern Spain. Calculation of the energy (glucose) required for their synthesis was based on the quantification of chemical components present in tissues. Despite considerable differences in the chemical composition of the three life forms studied (trees, shrubs and herbaceous), detected differences in construction cost were non‐significant (mean value: 1·64 ± 0·13 g glucose g^?1). However, shrubs and herbaceous plants growing in more fertile habitats expended significantly less energy on root synthesis (1·58 ± 0·06 and 1·41 ± 0·05 g glucose g^?1, respectively) than those growing in less fertile areas (1·80 ± 0·06 and 1·57 ± 0·1 g glucose g^?1, respectively), because they contained smaller amounts of either waxes (shrubs) or lignins (herbaceous), both expensive to synthesize, and, proportionately, more cellulose; which is inexpensive to synthesize. Deciduous and evergreen tree species also differed mainly with regard to wax and cellulose contents, giving rise to a significantly higher construction cost in evergreens (1·57 ± 0·07 g glucose g^?1 versus 1·78 ± 0·02 g glucose g^?1). The differences observed in construction cost appeared to be due more to habitat‐induced differences in chemical composition than to any intrinsic difference between the species studied.     

Alkaline opening of imidazole ring of 7-methylguanosine. 2. Further studies on reaction mechanisms and products

High performance liquid chromatography (HPLC) was used to follow the kinetics of the alkaline induced opening of the imidazole ring of 7-methylguanosine (7-meGuo). The kinetics show an initial rapid formation of a major transient intermediate and some minor products that were chromato-graphically separable into seven peaks. This phase of the reaction is followed by the formation of a dominant pyrimidine derivative whose liquid chromatography retention time in a 6% methanol, 0.01 M NH₄H₂PO₄ (pH 5.1) solvent is 6 min; during the rest of the reaction time this dominant species was progressively converted to a co-dominant species that has a 4.5-min column retention. Mass spectroscopy confirmed the existence of two species of ring opened 7-methylguanine (7-meGua), one formylated and another deformylated. Schiff's reaction demonstrated that the species in the second HPLC peak is the formylated one. The ring opened 7-methylguanine (rom⁷Gua) released by formamidopyrimidine (FAPy)-DNA glycosylase was shown to coelute with the formylated species. These results demonstrate that the enzyme excises formylated rom⁷Gua from DNA Analysis of rom⁷Guo by NMR showed that there are two signals assignable to methyl protons and two to formyl protons. These chemical shifts were interpreted as being due to the opening of the imidazole ring at two sites and to the formation of formylated and deformylated rom⁷Gua.     

Seasonal Variation in the Chemical Composition of Tropical Australian Marine Macroalgae

The proximate chemical composition (ash, soluble carbohydrate, lipid and protein) was determined in 30 common species of tropical Australian marine macroalgae from Darwin Harbour (12^∘26′S, 130^∘51′E), in summer (hot and wet) and winter (cool and dry). There was a wide diversity of species in both seasons (19 species in summer and 20 species in winter). In most species, the major component was soluble carbohydrate (chlorophytes range 2.5–25.8% dry weight (dw), phaeophytes range 8.4–22.2% dw, rhodophytes range 18.7–39.2% dw) with significantly higher (p < 0.05) percentages only in winter season rhodophytes. Highest percentages of protein were found in rhodophytes collected in the summer (range 4.8–12.8% dw), with significantly lower percentages (p < 0.05) during winter. All species had lipid contents within the range 1.3–7.8% dw, with highest percentages in summer phaeophytes, but no significant differences between species or season. Most species had moderate to high ash contents (24.2–89.7% dw), with the highest percentages during summer. Compared with summer samples, macroalgae collected in winter had higher energy value and slightly lower percentages of inorganic matter. The variation of algal groups and chemical composition may influence the availability of the food source for the majority of herbivores, which in turn is likely to effect their ecology and community structure.     

Combined gas chromatography-chemical ionization mass spectrometry of sphingolipids. I. Glucosyl sphingosine,ceramides and cerebrosides of the spleen in Gaucher's disease

Trimethylsilylated glucosyl sphingosine, ceramides and glucocerebrosides were analysed by combined gas chromatography (GC)-chemical ionization (CI) mass spectrometry. Isobutane, methane and ammonia were used as reactant gases for chemical ionization. Essentially the same fragment ions were detected in the spectra with the different reactant gases.Purified glucocerebrosides isolated from the spleen of a patient with Gaucher's disease were clearly separated into their 8 molecular species by gas chromatography. Three other minor components were detected by spectrometry, and these 11 molecular species of glucocerebrosides from the spleen of the patient with Gaucher's disease have been analysed.The ceramides obtained by periodate oxidation and then alkaline reduction of the glucocerebrosides were also separated into 11 molecular species by GC-CI mass spectrometry.Molecular weight could be determined using the major fragment ion of m/e (M⁺?90) in the spectra of ceramides and cerebrosides. The chemical ionization method is useful for structural analyses and determination of the molecular species of sphingoglycolipids.     

Chemical speciation of L-proline complexes of Ca(II), Zn(II) and Mn(II) in acetonitrile-water mixtures

Abstract

Chemical speciation of binary complexes of Ca(II), Zn(II) and Mn(II) with L-proline is investigated pH-metrically in acetonitrile-water mixtures. The stability constants are calculated using the computer program MINIQUAD75. The best-fit chemical models are selected based on statistical parameters and residual analysis. The models for the binary species contained ML⁺, MLH²⁺and ML₂H⁺ for Ca(II), Zn(II) and Mn(II). The trend in variation of stability constants with change in the dielectric constant of the medium is explained on the basis of structure forming nature of acetonitrile. Distribution of the species with pH at different variations (0.0-60.0% v/v) in acetonitrile-water mixtures is also presented.     

Physical and chemical effects of litter on plant establishment in semi-natural grasslands

A field experiment was conducted in a semi-natural grassland to study the interspecific variation in the effect of litter on seedling emergence and establishment and separate physical from chemical effects. Seeds of seven forb species were sown in plots subjected to either litter amendment (0, 400 or 900 g m⁻²) or water extracts of litter (corresponding to 400 and 900 g litter m⁻²). In addition, an extract was treated with activated carbon to estimate the possible effects of secondary chemical compounds. The response to plant litter differed amongst species: negative, neutral and positive responses were observed. Anthriscus sylvestris was the only species with a strong positive response to litter. We found no consistent relation between seed size and response to plant litter. Physical effects of litter were generally stronger than chemical effects. However, water extract of litter inhibited emergence in three species. Activated carbon removed the negative effect of the litter extract, which suggests that the effect was caused by an inhibitory chemical compound rather than by increased competition in response to nutrients added via the extract. The balance between facilitative and inhibitory effects of litter depended on species identity and litter quantity. Facilitative effects dominated at low and intermediate quantities of litter, and inhibitory effects at high litter quantities. One species, Campanula rotundifolia, showed a switch from positive to negative responses with increasing quantities of litter. However, we found no general threshold for litter quantity valid across species.     

Speciation of zinc and tin(II) in dentrifices with reference to availability in saliva

Abstract

The chemical speciation of zinc- and tin(II)-containing dentrifices and of the reaction between saliva and such toothpastes has been researched using computer simulation. Pivotal formulation variables have been identified (phosphates, fluorides, citrate, etc.) and optimised to suggest formulations having maximum availability, of antimicrobial metal salts Zn²⁺ (aq) and soluble tin(II) species. Although the concept of having a single dentifrice in which both the zinc and the soluble tin(II) species are maximised together is desirable, it is concluded that at a single pH value, there cannot be maximal availability for both species, but it is possible to achieve levels of antimicrobial metal salts superior to a dentifrice containing either ingredient alone.     

NO• chemistry: a diversity of targets in the cell

Abstract

NO^? alone is a poorly reactive species; however, it is able to undergo secondary reactions to form highly oxidizing and nitrating species, NO₂^?, N₂O₃, and ONOO^?. These secondary reactive nitrogen species (RNS) are capable of modifying a diversity of biomolecular structures in the cell. The chemical properties of individual RNS will be discussed, along with their ability to react with amino acids, metal cofactors, lipids, cholesterol, and DNA bases and sugars. Many of the identified RNS-induced modifications have been observed both in vitro and in vivo. Several of these chemical modifications have been attributed with a functional role in the cell, such as the modulation of enzyme activity. Other areas in the field will be discussed, including the ability of RNS to react with metabolites, RNA, and substrates in the mitochondrion, and the cellular removal/repair of RNS-modified structures.     

Evenness of soil organic carbon chemical components changes with tree species richness,composition and functional diversity across forests in China

Higher tree species richness generally increases the storage of soil organic carbon (SOC). However, less attention is paid to the influence of varied tree species composition on SOC storage. Recently, the perspectives for the stronger persistence of SOC caused by the higher molecular diversity of organic compounds were proposed. Therefore, the influences of tree species richness and composition on the molecular diversity of SOC need to be explored. In this study, an index of the evenness of diverse SOC chemical components was proposed to represent the potential resistance of SOC to decomposition under disturbances. Six natural forest types were selected encompassing a diversity gradient, ranging from cold temperate to tropical forests. We examined the correlations of tree species richness, composition, and functional diversity, with the evenness of SOC chemical components at a molecular level by ¹³C nuclear magnetic resonance. Across the range, tree species richness correlated to the evenness of SOC chemical components through tree species composition. The negative correlation of evenness of SOC chemical components with tree species composition, and the positive correlation of evenness of SOC chemical components with tree functional diversity were found. These indicate the larger difference in tree species composition and the lower community functional diversity resulted in the higher heterogeneity of SOC chemical components among the communities. The positive correlation of the evenness of SOC chemical components with the important value of indicator tree species, further revealed the specific tree species contributing to the higher evenness of SOC chemical components in each forest type. Soil fungal and bacterial α-diversity had effect on the evenness of SOC chemical components. These findings suggest that the indicator tree species conservation might be preferrable to simply increasing tree species richness, for enhancing the potential resistance of SOC to decomposition.     

Tree Foliar Chemistry in an African Savanna and Its Relation to Life History Strategies and Environmental Filters

Understanding the relative importance of environment and life history strategies in determining leaf chemical traits remains a key objective of plant ecology. We assessed 20 foliar chemical properties among 12 African savanna woody plant species and their relation to environmental variables (hillslope position, precipitation, geology) and two functional traits (thorn type and seed dispersal mechanism). We found that combinations of six leaf chemical traits (lignin, hemi-cellulose, zinc, boron, magnesium, and manganese) predicted the species with 91% accuracy. Hillslope position, precipitation, and geology accounted for only 12% of the total variance in these six chemical traits. However, thorn type and seed dispersal mechanism accounted for 46% of variance in these chemical traits. The physically defended species had the highest concentrations of hemi-cellulose and boron. Species without physical defense had the highest lignin content if dispersed by vertebrates, but threefold lower lignin content if dispersed by wind. One of the most abundant woody species in southern Africa, Colophospermum mopane, was found to have the highest foliar concentrations of zinc, phosphorus, and δ¹³C, suggesting that zinc chelation may be used by this species to bind metallic toxins and increase uptake of soil phosphorus. Across all studied species, taxonomy and physical traits accounted for the majority of variability in leaf chemistry.     

1H, 15N and 13C resonance assignments of yeast Saccharomyces cerevisiae calmodulin in the Ca2+-free state

Calmodulin (CaM) is a small Ca²⁺-binding protein, which has been found in all of eucaryotic cells examined. CaMs isolated from various species have highly conserved amino acid sequence (more than 90% identical), and show the same biological functions. CaM isolated from the baker's yeast (Saccharomyces cerevisiae) (yCaM), however, shares only 60% identity in the amino acid sequence with CaM from vertebrate, and shows quite distinct conformational and biochemical properties compared with those of CaM from other species. The conformational details of yCaM, however, have not been revealed yet. We achieved the chemical shift assignments of yCaM (146 amino acids) in the apo-state using uniformly ¹⁵N- and ¹³C-labeled protein. Consequently, the resonances of 95% atoms in the backbone amides were successfully assigned.     

THE BEGINNINGS OF ANTARCTIC MACROALGAL CHEMICAL ECOLOGY: DEFENSES AGAINST HERBIVORES IN A NITROGEN REPLETE,CARBON LIMITED OCEAN

Amsler, C. D.¹, Iken, K. B.¹, McClintock, J. B.¹, Furrow, F. B.², & Baker, B. J.^{2 1}Department of Biology, University of Alabama at Birmingham, Birmingham AL 35294-1170 USA; ²Department of Chemistry, Florida Institute of Technology, Melbourne, FL 32901 USA We examined palatability of Antarctic Peninsula macroalgae in feeding bioassays with three common, sympatric macroalgal-consuming omnivores (amphipod, fish, sea star). Antarctic macroalgae have low C:N ratios, high nitrogen contents, and are usually growth limited by carbon (light). The Carbon Nutrient Balance Hypothesis (CNBH) predicts that macroalgae would produce nitrogenous secondary metabolites for defense rather than high levels of non-nitrogenous defenses under these conditions. To date, feeding bioassays have been performed on fragments of thallus from 26 macroalgal species and 21 (81%) were rejected by at least one omnivore. Organic extracts from 13 macroalgal species rejected as thallus were used in feeding bioassays. At least one extract from 12 species (92%) was rejected by at least one omnivore, suggesting that chemical defenses against herbivores probably are present in at least some of the macroalgal species. We have identified a number of specific, non-nitrogenous secondary metabolites in these extracts and previous workers have also reported non-nitrogenous secondary metabolites from antarctic macroalgae. Additional extracts targeting nitrogenous metabolites from 25 species were subjected to thin layer chromatography and visualized by stains specific for nitrogenous compounds. No nitrogenous secondary metabolites were identified by this or other methods. Nitrogenous secondary metabolites are also extremely rare in macroalgae from other areas of the world. Consequently, although our bioassays suggest that chemical defenses probably do occur, our data cast doubt on the applicability of the CNBH for predicting the chemical composition of macroalgal defenses under carbon limited conditions. (NSF OPP9814538, OPP9901076)     

Contents to volume 155

The incorporation of two fluorine-containing general anesthetic agents, halothane and methoxyflurane, into erythrocytes (from three different species), rabbit muscle and rabbit nerve, was followed with ¹⁹F NMR spectroscopy. Two major findings emerged from these studies: (1) multiple environments indicative of domain structure in the membrane can be observed depending on the anesthetic and the tissue type; and (2) the ¹⁹F chemical shifts of a given anesthetic were characteristic for the tissue examined. Halothane showed a single resonance in erythrocytes and multiple resonances in muscle and nerve, while methoxyflurane showed multiple resonances in both muscle and erythrocytes. The range of the ¹⁹F chemical shifts for the multiple peaks was as great as 6 ppm.