Effects of inorganic arsenicals on DNA synthesis in unsensitized human blood lymphocytes in vitro

Effects of inorganic arsenicals on DNA synthesis in unsensitized human blood lymphocytes were biphasic: The chemicals at very low concentrations enhanced DNA synthesis, whereas higher concentrations inhibited DNA synthesis. The concentrations of arsenicals at which the maximum stimulating effect was found were 1×10^?5 M, 1×10^?6 or 2×10^?6 M, and 0.8×10^?6 or 1×10^?6 M for sodium arsenite exposure of 1 h, 3 d, and 6 d, respectively; for sodium arsenate, 1× 10^?5 M, 1×10^?5 M, and 2×10^?6 or 5×10^?6 M, respectively. Arsenicals must be present for the entire 6-d culture period to produce maximum stimulation of DNA synthesis in human lymphocytes. The longer exposure of the lymphocytes to arsenicals, the lower the concentrations of arsenicals at which the maximum stimulating effect on DNA synthesis was found. Stimulating effect of trivalent arsenic (sodium arsenite) on DNA synthesis was stronger than pentavalent arsenic (sodium arsenate), and the stronger the effect of trivalent arsenic than pentavalent, the longer exposure of the cells to the chemicals. Both sodium arsenite and sodium arsenate stimulated DNA synthesis in human lymphocytes to a lower degree than phytohemagglutinin (PHA).     

Production of ethylene,its precursors and implied enzyme activities in isolated chickpea embryonic axes during the onset of growth

The embryonic axes of chickpea (Cicer arietinum) seeds were used to quantify 1-(malonyl)aminocyclopropane-1-carboxylic acid (MACC), 1-aminocyclopropane-1-carboxylic acid (ACC), ethylene and some related enzymes during the initial 18 h imbibition period (anaerobic growth phase). Longer cold storage (stratification) of seeds produced higher levels of MACC and ACC. Maximum accumulation of MACC and malonyl-transferase activity occurred after 5 h of growth but MACC levels later became insignificant. ACC-synthase activity and endogenous ACC seem to reach a maximum 2 h after MACC accumulation. MACC-hydrolase activity was measured“in-vitro” and reached a maximum after 5.5 h of growth. These results suggest that endogenous MACC did not seem to be an end product; it may be involved in ACC production and the regulation of ethylene production before the emergence of the radicle. Ethylene-forming enzyme (EFE) activity reached a maximum after 12 h and ethylene production after 18 h of growth. The physiological implications of this temporal separation of MACC, ACC, ethylene and related enzymes is discussed.     

Hormone-like product of vascular tissue stimulating starch accumulation in pith explants of kale and endogenous cytokinins

When stem explants of kale (Brassica oleracea L. var.medullosa), containing pith parenchyma and a strip of vascular tissue, were cultured on simple sucrose medium, a hormone-like factor was transported from the vascular tissue to the adjacent pith, where it stimulated accumulation of starch. Similarly, up to a sevenfold increase of starch content in explants could be induced by cytokinins added to the culture medium. The relative stimulatory effect of several cytokinins (5×10^?6 M) and hormone-like product of vascular tissue (HPVT) in a typical experiment were: control (1.0), trans-zeatin (6.7), HPVT (6.2), N⁶-[2-isopentenyl]adenine (5.4), transzeatin riboside (5.2), N⁶-[2-isopentenyl]adenosine (5.4), kinetin (3.6), 6-benzylaminopurine (3.5), and adenine (2.1). Concentration of endogenous cytokinins was determined using ELISA (trans-zeatin, N⁶-[2-isopentenyl]adenine and their ribosides) andAmaranthus bioassay (total cytokinins). No effect of vascular tissue on the level of endogenous cytokinins in explants was found. The results support the conclusions of previous experiments that the HPVT stimulating starch accumulation is not a cytokinin.     

Superoxide dismutase level in response to paraquat and high temperature in the cyanobacteriumGloeocapsa sp

Paraquat (methylviologen) at concentrations above 0.05 mM inhibited the growth of photoautotrophic cyanobacteriumGleeocapsa sp. in axenic cultures. The growth rate was not affected by concentrations of 0.01 mM or less. This concentration resulted after a lag period in a moderate increase in superoxide dismutase level. After removal of paraquat, the cyanobacterium continued to generate higher levels of superoxide dismutase. There was a lag period of one hour before resumption of normal enzyme activity. Addition of puromycin at concentration of 0.5 mg cm^?3 had no effect on cell survival, but greatly enhanced the sensitivity of the culture to the toxicity of paraquat. The data showed an increase of SOD activity by temperatures above the normal growth temperature level. However, this increase was suppressed by chloramphenicol which revealed that the induction of superoxide dismutase by high temperatures was associated withde novo protein synthesis.     

The combined effects of acidification stress and kinetin on chlorophyll content,dry matter accumulation and transpiration coefficient inSorghum bicolor plants

Increasing soil acidity (from pH 6.5 to pH 2.0) decreased chlorophyll (Chl)a andb contents, dry matter accumulation by plants and the transpiration coefficient. Chl stability to heat significantly increased with increased soil acidity. The Chla/b ratio was increased significantly at pH 5 and 4 and decreased at pH 3 and 2. SprayingSorghum shoots with kinetin solutions counteracted the above adverse effects on Chl content and dry matter accumulation. Kinetin-treated plants showed a lower transpiration coefficient than the untreated plants.     

Trace elements nutritional status

In this study, hair levels of Cu and Zn were determined in healthy male and female individuals (n=192) ages 3.6–14.5 yr and the correlations with Cu and Zn daily intakes were examined. Determinations of Cu and Zn concentrations were performed by way of atomic absorption spectrometry and X-ray fluorescence. Nutritional data were collected with the aid of an individual questionnaire. Statistical analysis revealed no effect of age and sex either on Cu concentrations in hair or on Cu daily intakes. Zn concentrations were significantly higher in hair of both pubescent males and females compared with prepubescent individuals. There was no influence of age on Zn daily intake in males, however, whereas pubescent girls had a lower intake than males. Correlation coefficients between Cu concentrations in hair and daily nutritional intakes calculated for males and females werer=0.1694 andr=0.1677, respectively; those for Zn werer=?0.2223 (p<0.05) in males andr=?0.2787 (p<0.01) in females. These data confirm that the analysis of Zn in hair represents an addition to conventional materials in the assessment of the nutritional status of groups of individuals.     

Enhancement of faba bean nodulation,nitrogen fixation and growth by different microorganisms

The effect of four bacterial and six fungal species on nodulation and growth ofVicia faba cv. Giza 3 inoculated withRhizobium leguminosarum biovarviceae strain RCR 1001 were assessed in a pot experiment.Bacillus cereus, Pseudomonas fluorescens, Aspergillus niger andA. quadriliniatus either alive cells or sterile filtrate significantly promoted nodulation, growth and nitrogen accumulation. Heat-killed cells had no effect.     

Effect of Mg,Zn and Mo salts on nitrate reductase activity and soluble protein content in leaves ofQuercus serrata

Leaves ofQuercus serrata seedlings were sprayed with solutions of different concentrations of Mg, Zn and Mo-salts and activity of nitrate reductase (NRA) and soluble protein contents were determined in fresh leaves of different ages. The optimum dose for magnesium sulphate and zinc sulphate was 2.5 mM while for ammonium molybdate it was 0.01 mM. Under optimum doses of these nutrients high levels of NRA and protein contents were maintained upto 28 d of spray. Observations on NRA were further confirmed by leaf direct treatment and excised shoot-dipping experiments which showed maximum NRA at comparatively much lower concentrations of ions than those used for spray.     

Cytological relationships of selected species ofPanicum L.

The cytological investigation of 12 taxa ofPanicum L. revealed that the vast majority of them have the basic number x=9 at different ploidy levels. The basic number x=8 was recorded only in the tetraploid speciesP. maximum with 2n=32. The diploid number 2n=18 was encountered inP. capillare, P. laevifolium, P. antidotale andP. coloratum (2) with 3B-chromosomes recorded in the latter species. The tetraploid chromosome number 2n=36 was found to exist inP. miliaceum, P. miliare, P. coloratum (1) andP. virgatum. The hexaploid number 2n=54 was recorded inP. bulbosum, P. dichotomiflorum andP. esculentum. The karyotypes of all accessions were mostly symmetrical and mainly comprised of meta- and submetacentric chromosomes with little variation in length among them within each karyotype. Investigation of chromosome association during metaphase I of meiosis revealed that the frequency of bivalents/cell was the highest among all investigated diploid, tetraploid and hexaploid accessions. Univalents were also frequently encountered in various accessions. These results may indicate that segmental alloploidy has been the major process by which polyploid species have originated.     

Characterization of mitochondrial DNA topoisomerase I fromNeurospora crassa

DNA topoisomeraseI isolated from the lower eukaryoteNeurospora crassa mitochondria was characterized. Molar mass of the enzyme in the native state is 120 kDa and 60–65 kDa when denatured. The pH optimum of the enzyme is 7.8 and the KCl optimum concentration is 40 mmol/L. This topoisomerase is independent of ATP and Mg²⁺. N-Ethylmaleimide, 4-chloromercuribenzoate, SDS, guanidinium chloride, polyethylene glycol, heparin and ethidium bromide inhibit its activity, while novobiocin, nalidixic acid, Triton X-100 and chloroquine do not. Polyamines, and histone, H1 stimulate the topoisomerase activity. We classify this DNA topoisomerase as typeI and eukaryotic. Conversion of the topoisomerase to a nonspecific endonuclease at increased temperature is proposed.     

Identification and application of amino acids as chelators in phytoremediation of rare earth elements lanthanum and yttrium

Aims

This study aimed to identify amino acids that could act as chelators in enhancing absorption and translocation capabilities of a rare earth element (REE) lanthanum and yttrium in a non-hyperaccumulator plant.

Methods

We analysed correlations between amino acid, La and Y concentrations in xylem saps of the REE hyperaccumulator Phytolacca americana L., to identify functional amino acids in long-distance transportation processes. These were used as chelators to observe the efficacy of La uptake and translocation in REE non-hyperaccumulator tomato seedlings. Pot culture experiments were conducted using modified Hoagland solution artificially contaminated with REEs.

Results

Eighteen xylem sap amino acids were identified and measured in the xylem sap, using reversed-phase high-performance liquid chromatography. Aspartic acid, asparagine, histidine and glutamic acid may be related to xylem La and Y long-distance transportion in P. americana L. Extraneous aspartic acid, asparagine, histidine and glutamic acid enhanced La absorption in the whole tomato, compared with La alone. Moreover, the whole tomato La content increased by 449 μg and 139 μg in the presence of aspartic acid and asparagine as compared to P. americana L. Tryptophan insignificantly affected La uptake and translocation in tomato seedlings.

Conclusions

Aspartic acid and asparagine could potentially promote remediation of La contamination in soil when used as a chelator.     

Comparison of twoPisum sativum nodulation mutants with their parental cultivar

In comparison with the parental cv. Finale the ‘RisfixC’ supernodulator exhibited higher, continuously increasing nodule number and fresh mass accumulation, but substantially lower individual nodule fresh mass, leghemoglobin concentration, and specific acetylene reduction activity of nodule tissue. There were no substantial differences between Finale and ‘RisfixC’ in total acetylene reduction, nodule leghemoglobin accumulation per nodulated root, total and specific CO₂ evolution from nodulated roots and gross CO₂ respiratory costs of acetylene reduction. The ‘RisfixC’ also exhibited a substantially lower plant dry mass production (by 30%), but nitrogen concentration in shoots and carotenoid concentration in leaf tissue were significantly higher by 33 and 14%, and the chlorophylla+b content insignficantly higher than in the parental cultivar. In contrast, the nodulation mutant ‘Risnod29’, exhibited a somewhat higher nodule fresh mass accumulation (by 21%) and individual nodule fresh mass (by 23%), total and specific acetylene reduction (by 49 and 19%) and a somewhat more rapid plant dry mass accumulation compared with the cv. Finale.     

Serum copper and zinc in industrial centers in Slovakia

The association between serum copper and zinc concentrations and age, sex, and other risk factors of cardiovascular disease in randomly selected adult volunteers aged 19–59 were investigated. There was a positive relationship between copper and age in both sexes, but zinc was negatively correlated with age in males only. Serum zinc was positively related to HDL-cholesterol in males. Serum copper was positively related to total cholesterol and LDL-cholesterol but negatively correlated to HDL-cholesterol in males. A positive relationship to body mass index was observed in females only. Subjects have been divided into a control group and a group with marked risk factors of cardiovascular disease. The levels of zinc were not different, whereas the levels of copper in both males and females were significantly higher in the risk group. Our results suggested a positive relationship between serum copper and cumulation of more factors of cardiovascular disease, however, their causal effect in humans has to be investigated further.     

Subcellular localization of ribonuclease isoenzymes in tobacco mesophyll protoplasts and their changes induced by infection of PVY

Changes in the content and subcellular localization of ribonuclease isoenzymes were determined in mesophyll protoplasts prepared fromNicotiana tabacum L. cv. Samsun from healthy and potato virus Y (PVY) infected plants. Intact chloroplasts, mitochondria and soluble cytosolic proteins were obtained after protoplast disintegration by means of differential centrifugation. The 1 000g pellet from healthy protoplasts contained 7.3 %, the 15 000g pellet 13.5 % and 15 000g supernatant 82.1 % of the total activity of ribonucleases. The 1 000g pellet from infected protoplasts contained 10.4%, the 15 000g pellet 10.0% and 15 000g supernatant 89.6 % of the total activity of ribonucleases. The activity of these enzymes in infected protoplasts was enhanced in crude homogenate to 137.0 % (P<0.001), in 1 000g pellet to 194.8 % (P<0.001), in 15 000g pellet to 101.3 % (NS), and in 15 000g supernatant to 149.4 % (P<0.001) of that in healthy noninoculated protoplasts.     

Plant radioresistance and DNA repair efficiency inChlamydomonas reinhardtii andPisum sativum

This paper compares the repair of DNA single strand breaks (ssb) induced by γ-radiation in two strains ofChlamydomonas reinhardtii (137C/+/ and UVS-I) and three lines ofPisum sativum (NN 131, 198, 140) differing in the degree of radioresistance. DNA ssb in cells exposed to γ-rays (50, 100, 200, 500 Gy) were measured by electrophoresis and alkaline unwinding method with subsequent chromatography on hydroxyapatite immediately after irradiation and after 30 min of post-irradiation incubation at 25°C. An increase of double-strand DNA (in%) was found in cells after 30 min post-irradiation incubation.C. reinhardtii strains displayed an equal level of DNA degradation and repair efficiency in the DNA single strand breaks. The radioresistant line N 198 ofP. sativum is characterized by a lower level of induced DNA ssb and higher efficiency of repair of these breaks as compared with less radioresistant lines NN 131 and 140.     

Alcohol dehydrogenase isoenzymes fromNicotiana tabacum include ADH of bothN. sylvestris andN. tomentosiformis

Electrophoretic separation of seed alcohol dehydrogenase (ADH) fromNicotiana tabacum on 12% starch gels at pH 7.8 produced only one band with an apparent R_f of 0.65, which confirmed earlier reports. The same was found with pollen ADH. However, in polyacrylamide gel isoelectric focusing, seed ADH separated into three distinct bands with apparent pI of 5.33, 5.42 and 5.50. The pI 5.33 isoenzyme was found to be the essential form inN. sylvestris seeds. The analysis of charge properties ofN. tomentosiformis seed ADH showed only one isoenzyme with pI of 5.56. These results present further evidence thatN. tabacum has arisen from a cross between aN. sylvestris predecessor and an ancestral type ofN. tomentosiformis. The presence of the pI 5.42 form inN. tabacum is consistent with the reported formation of heterodimeric ADH in tobacco hybrids.     

Measurement of the elemental composition of nasal-pharynx cancerous tissue with PIXE

Twently elements in two samples of nasal-pharynx cancerous tissue (NPCT) have been measured with PIXE. The results show that the element concentrations of calcium and zinc are very high, up to several thousand μg/g. Compared with results from other types of cancer patients, trace element concentrations in NPCT seem to be in a special status.     

Flow cytometric estimation of nuclear DNA amount in diploid bananas (Musa acuminata andM. balbisiana)

Cell nuclei were isolated from leaf tissues of wild banana (Musa balbisiana, M. acuminata ssp.banksii andM. acuminata ssp.errans) and of the two vegetative clones of diploid cultivar “Pisang Mas”. Relative fluorescence intensity was measured on propidium iodide-stained nuclei by flow cytometry. Nuclei isolated fromGlycine max with known nuclear genome size were used as internal standard to determine nuclear DNA content ofMusa in absolute units. The results of the study showed that the size of nuclear genome ofMusa is smaller than previously estimated. In general, it is smaller in comparison with many other angiosperms. Furthermore, it was found that nuclear DNA content ofM. balbisiana (genome BB) is significantly lower than that ofM. acuminata subspecies and cultivars (genome AA). This finding should permit estimation of genome composition in triploidMusa clones with expected hybrid composition. Flow cytometry is proposed as a useful technique with potential applications in taxonomy, breeding and biotechnology ofMusa.