Determination of the triacylglycerol composition of coffee beans by reverse-phase high-performance liquid chromatography

Reverse-phase HPLC with refractive index and light scattering detectors in isocratic and gradient elution modes, respectively, was applied for the separation of the major triacylglycerols (TAG) in coffee lipids. Twelve TAG species could be identified and determined using a linear gradient of acetonitrile in dichloromethane: dichloroethane. The quantitative evaluation was based on the relative area percentages derived directly from a data-station. The procedure was applied to determine the TAG composition of three types of coffee beans harvested in two coffee producing areas in Brazil and dried by two commonly used procedures. No significant differences in the TAG compositions due to the type, origin and drying procedure were found.     

Analysis of mtDNA indicates weak temporal genetic heterogeneity in pink salmon spawning runs in two rivers on Sakhalin Island

Both odd- and even-year pink salmon populations were sampled during the spawning runs in Firsovka and Bakhura rivers on Sakhalin Island. Four collections of 30 fish spaced at 2-week intervals were taken from each river in 2 consecutive years. Four restriction endonucleases were used to examine 2·15% of the mitochondrial genome. Eighteen variable sites and three types of insertion defined 37 haplotypes among 449 fish. Heterogeneity tests showed highly significant differences among temporal collections taken in the odd year from Bakhura River, and no significant differences among temporal collections from the other three spawning runs. However, probabilities of homogeneity among temporal collections were low in all tests, and an integral estimate of the probability of homogeneity for the total set of tests was less than 0·001, indicating highly significant overall temporal heterogeneity. Analysis of molecular variance (AMOVA), revealed that a small portion of the variance was distributed among temporal collections, and small φ_ST values that differed significantly from zero only in the odd-year population in Bakhura River. Temporal differences in spawning are probably adapative, and allow greater productivity in areas of high spawning densities.     

Suitability of flow cytometry for estimating bacterial biovolume in natural plankton samples: comparison with microscopy data

The relationship between flow cytometry data and epifluorescence microscopy measurements was assessed in bacterioplankton samples from 80 lakes to estimate bacterial biovolume and cell size distribution. The total counts of 4',6'-diamidino-2-phenylindole-stained cells estimated by both methods were significantly related, and the slope of their linear regression was not significantly different from 1, indicating that both methods produce very similar estimates of bacterial abundance. The relationships between side scatter (SSC) and 4',6'-diamidino-2-phenylindole fluorescence and cell volume (microscopy values) were improved by binning of the data in three frequency classes for each, but further increases in the number of classes did not improve these relationships. Side scatter was the best cell volume predictor, and significant relationships were observed between the SSC classes and the smallest (R2 = 0.545, P < 0.001, n = 80) and the largest (R2 = 0.544, P < 0.001, n = 80) microscopy bacterial-size classes. Based on these relationships, a reliable bacterial biomass estimation was obtained from the SSC frequency classes. Our study indicates that flow cytometry can be used to properly estimate bacterioplankton biovolume, with an accuracy similar to those of more time-consuming microscopy methods.     

Inducible and constitutive mechanisms of salt stress resistance in <Emphasis Type="Italic">Geum urbanum</Emphasis> L.

The avens (Geum urbanum L.) seedlings were grown for 6 weeks until the expansion of five to six leaves and then exposed to salinity shock (300 mM NaCl in the nutrient medium) or to a gradual (within 4 days) increase in NaCl concentration from 100 to 400 mM. The dynamics of stress-dependent accumulation of Na⁺, Cl^?, proline, and polyamines in leaves and roots was measured, together with activities of antioxidant enzymes, namely, superoxide dismutase (SOD) and guaiacol-dependent peroxidase occurring in soluble, ionically bound, and covalently bound forms. It is shown that avens plants can adapt to gradual salinization by mobilizing stressinducible protective mechanisms (accumulation of proline and spermine) and by activating constitutive enzyme systems (SOD and peroxidase).     

Phytoremediation potential of <Emphasis Type="Italic">Amaranthus</Emphasis> hybrids: Antagonism between nickel and iron and chelating role of polyamines

Three amaranth hybrids (Amaranthus paniculatus f. cruentus (Vishnevyi dzhem), A. paniculatus (Bronzovyi vek), and A. caudatus f. iridis (Izumrud) were grown in the climate-controlled chamber on Jonson nutrient medium supplemented with 2 μM Fe³⁺-EDTA. When plants developed 5–6 true leaves (six-week-old plants), NiCl₂ was added to medium to final concentrations of 0 (control), 50, 100, 150, 200, and 250 μM. In 6 days, the increment in biomass of young and mature leaves, stems, and roots, and also the contents of Ni and Fe in them were measured. The red leaf amaranth hybrid Vishnevyi dzhem manifested the highest phytoremediation potential. i.e., the highest capacity for Ni accumulation in the shoots and the most pronounced symptoms of Fe deficit. In the presence of 150 and 250 μM NiCl₂ in medium, the shoots of these plants contained about 2 and 4 mg Ni/g dry wt, respectively. In experiments with Fe deficit in plants grown for a week in the presence of NiCl₂ (0, 25, 50, 75, and 100 μM), it was established that all tested nickel concentrations suppressed iron reduction in intact roots, which is catalyzed by ferric-chelate reductase, and this may underlie the antagonism between the two metals. In the presence of 50 μM NiCl₂ in medium and 2 μM Fe³⁺ (Fe deficit) and especially 100 μM Fe³⁺ (Fe excess), the content of MDA and proline in leaves increased and superoxide dismutase was activated; this indicates a development of oxidative stress. Leaf treatment with polyamines (putrescine or spermidine) with aminoguanidine (the inhibitor of H₂O₂ generation at polyamine oxidation) and with 1,3-diaminopropane led to the increase in nickel accumulation in leaves but did not result in the appearance of any signs of injury. This confirms our previous suggestion that polyamines manifest their protectory action as Ni chelators and detoxicants.     

The international society for plant anaerobiosis and its role in opening a new avenue of research

This paper is dedicated to the 35 year jubilee of the founding and the activity of the International Society for Plant Anaerobiosis (ISPA). The role of ISPA members in opening new avenues of research is emphasized. Major developments in the study of plant hypoxic and anoxic stress achieved during subsequent decades are considered. Special attention is given to plant adaptation and damage under conditions of oxygen deficiency and complete absence of oxygen as well as during the post-anaerobic period. Plant metabolic adaptation to anaerobic stress and the capacity of some plants to avoid anaerobiosis are discussed.     

Nickel accumulation in rape shoots (Brassica napus L.) increased by putrescine

Application of exogenous putrescine (Put) increases nickel accumulation in rape shoots (Brassica napus), improving potential for phytoremediation of contaminated soils. Plants were grown within a growth chamber in water culture for five weeks, then 250-500 microM NiCl2 was added to rooting media. Within 5 days of treatment, damaging effects of nickel manifested as a reduction of root system size and a decrease of Cu and especially Fe content in young leaves. Spraying leaves of adult plants with Put markedly reduced toxic effects of Ni on root growth, enhanced leaf supply with Fe, and increased Ni content in young leaves by 2.5 times. Plant growth in medium with elevated levels of Ni stimulated accumulation of endogenous spermidine (Spd), spermine (Spm), and especially Put (by 4 times as compared with the control). Results suggest that Ni-induced accumulation of endogenous polyamines in rape leaves is caused by activation of long-distance metal transport within the plant and reduction of its toxicity due to Put chelating action.     

The role of natural biopolymers in genotoxicity of mutagens/carcinogens elimination

Nowadays naturally occurring compounds with the potential antimutagenic and anticarcinogenic effects are of great importance for their prospective use in cancer chemoprevention and treatment. The new water soluble derivative of microbial polysaccharide beta-D-glucan-carboxymethyl glucan (CMG) belongs to such a category of natural substances. CMG isolated from the cell wall of baker's yeast Saccharomyces cerevisiae is included into the class of biopolymers known as biological response modifiers (BRMs) with a broad range of activities, above all ones interfering with cancer therapy. It was demonstrated on four experimental model systems that biological and consequential medicinal importance of CMG is based on the combined application with another active compound. In the Saccharomyces cerevisiae antimutagenicity assay CMG significantly reduced ofloxacin-induced mutagenicity in the yeast strain D7. CMG exerted bioprotective (anti-toxic and antimutagenic) effect after its simultaneos application with methyl methanesulphonate on the repair-deficient strain uvs10 of the unicellular green alga Chlamydomonas reinhardtii. In the Vicia sativa simultaneous phytotoxicity and anticlastogenicity assay CMG exerted statistically significant anticlastogenic efect against maleic hydrazide-induced clastogenicity in Vicia sativa L. Only in the Salmonella/microsome assay CMG did not exert statistically significant antigenotoxic effect, despite of the fact that it reduced 9-aminoacridine-induced mutagenicity in S. typhimurium TA97, but his(+) revertants decreasing was statistically significant only at the highest CMG concentration used. The data presented unambiguously documented that even biopolysaccharides (e.g., derivatives of beta-glucan) belonging to the most abundant class of natural biopolymers may contribute to cancer prevention and therapy.