The relationship between the levels of DNA-hydrocarbon adducts and the formation of sister-chromatid exchanges in Chinese hamster ovary cells treated with derivatives of polycyclic aromatic hydrocarbons

The frequencies of the induction of sister-chromatid exchanges and the levels of deoxyribonucleoside-hydrocarbon adducts formed in Chinese hamster ovary cells that had been treated with either dihydrodiols or a diol-epoxide derived from polycyclic aromatic hydrocarbons were determined. Up to 6-fold increases in the incidence of these exchanges were observed when the cells were treated either with the dihydrodiols, trans-3,4-dihydro-3,4-dihydroxy-7-methylbenz[a]anthracene,trans-7,8-dihydro-7,8-dihydroxybenzo[a]pyrene or the diol-epoxide, (±)-r-7, t-8dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a] pyrene but when the cells were transferred to media free of these compounds, there were rapid reductions in the frequency of these exchanges. When the exchanges were induced by the diol-epoxide, the decreases in frequency were paralleled by decreases in the levels of deoxyribonucleoside-diol-epoxide adducts that were present in hydrolysates of DNA isolated from the cells. There thus appears to be a close relationship between the frequency of sister-chromatid exchanges and the levels of deoxyribonucleoside-diol-epoxide adduct formation.     

Action Spectra for Conversions of Phycochrome c from Nostoc muscorum

The reversibly photochromic pigment, phycochrome c, was extracted from the blue-green alga Nostoc muscorum strain A. Action spectra were determined for in vitro conversions of the pigment from the short wavelength to the long wavelength form and vice versa. The action peak for the absorbance decrease at 650 nm is at 630 nm. During this decrease there is only a slight increase of the absorbance in the green region. Green and yellow light (maximum efficiency at 580 nm) completely restores absorbance at 650 nm. The observations are explained by the existence of three spectrally different forms of phycochrome c: P^c₆₃₀ and P^c₆₅₀ which equilibrate in darkness and P^c₅₈₀ which is reversibly photoconvertible to P^c₆₃₀. We have also measured the absorbance changes brought about by saturating irradiations with light of various wavelengths (“photostationary state spectrum”). Extreme photostationary states were obtained with about 650 nm and 500 nm light.     

Control of cell volume in the J774 macrophage by microtubule disassembly and cyclic AMP

We have explored the possibilities that cell volume is regulated by the status of microtubule assembly and cyclic AMP metabolism and may be coordinated with shape change. Treatment of J774.2 mouse macrophages with colchicine caused rapid microtubule disassembly and was associated with a striking increase (from 15-20 to more than 90 percent) in the proportion of cells with a large protuberance at one pole. This provided a simple experimental system in which shape changes occurred in virtually an entire cell population in suspension. Parallel changes in cell volume could then be quantified by isotope dilution techniques. We found that the shape change caused by colchicine was accompanied by a decrease in cell volume of approximately 20 percent. Nocodozole, but not lumicolchicine, caused identical changes in both cell shape and cell volume. The volume loss was not due to cell lysis nor to inhibition of pinocytosis. The mechanism of volume loss was also examined. Colchicine induced a small but reproducible increase in activity of the ouabain-sensitive Na(+), K(+)-dependent ATPase. However, inhibition of this enzyme/transport system by ouabain did not change cell volume nor did it block the colchicines-induced decrease in volume. One the other hand, SITS (4’acetamido, 4-isothiocyano 2,2’ disulfonic acid stilbene), an inhibitor of anion transport, inhibited the effects of colchicines, thus suggesting a role for an anion transport system in cell volume regulation. Because colchicine is known to activate adenylate cyclase in several systems and because cell shape changes are often induced by hormones that elevate cyclic AMP, we also examined the effects of cyclic AMP on cell volume. Agents that act to increase syclic AMP (cholera toxin, which activates adenylate cyclase; IBMX, and inhibitor of phosphodiesterase; and dibutyryl cyclic AMP) all caused a volume decrease comparable to that of colchicine. To define the effective metabolic pathway, we studied two mutants of J774.2, one deficient in adenylate cyclase and the other exhibiting markedly reduced activity of cyclic AMP-dependent protein kinase. Cholera toxin did not produce a volume change in either mutant. Cyclic AMP produced a decrease in the cyclase-deficient line comparable to that in wild type, but did not cause a volume change in the kinase- deficient line. This analysis established separate roles for cyclic AMP and colchicine. The volume decrease induced by cyclic AMP requires the action of a cyclic AMP-dependent protein kinase. Colchicine, on the other hand, induced a comparable volume change in both mutants and wild type, and thus does not require the kinase.     

Developmental stability in brown trout: are there any effects of heterozygosity or environmental stress?

We studied the developmental stability of brown trout, Salmo trutta L., in 10 populations (five acidified, five control) in Norway, measured as fluctuating asymmetry (FA) and departure from the morphological norm. We measured four meristic and four morphometric characters, and scored the level of biochemical heterozygosity at 49 loci (20 polymorphic). We reared eggs of a single population in a hatchery using four different water qualities (three replicates of each treatment) to test the effect of acidification stress on developmental instability. There were no significant differences in the level of FA, in departure from the morphological norm between brown trout sampled from lakes with acidified or control water qualities, or in brown trout hatched at different water qualities. There was no correlation between level of heterozygosity and FA or departure from the morphological norm, either when tested within populations or among populations. There were no single-locus effects on developmental stability tested for 11 loci. We conclude that measures of developmental stability or morphological variability are not useful for detecting acidification stress in brown trout. Furthermore, we conclude that developmental stability in our material varies independently of heterozygosity.     

Some studies on characterization of three phase partitioned chitosan

Three phase partitioning (TPP) is generally carried out by adding ammonium sulfate and t-butanol to a solution of a macromolecule. Chitosan could be obtained as an interfacial precipitate with 88% yield by subjecting 0.2% (w/v) chitosan solution to TPP with 45% (w/v) ammonium sulfate, with an equal volume of t-butanol at 40 °C. TPP resulted in structural changes which could be seen in its UV spectra, FT-IR spectra and solubility characteristics. TPP-treated chitosan also showed decreased susceptibility towards hydrolysis by chitinase. Thus, TPP can be used as a useful way of altering the properties of chitosan.     

A novel disorder caused by defective biosynthesis of N-linked oligosaccharides due to glucosidase I deficiency

Glucosidase I is an important enzyme in N-linked glycoprotein processing, removing specifically distal alpha-1,2-linked glucose from the Glc3Man9GlcNAc2 precursor after its en bloc transfer from dolichyl diphosphate to a nascent polypeptide chain in the endoplasmic reticulum. We have identified a glucosidase I defect in a neonate with severe generalized hypotonia and dysmorphic features. The clinical course was progressive and was characterized by the occurrence of hepatomegaly, hypoventilation, feeding problems, seizures, and fatal outcome at age 74 d. The accumulation of the tetrasaccharide Glc(alpha1-2)Glc(alpha1-3)Glc(alpha1-3)Man in the patient's urine indicated a glycosylation disorder. Enzymological studies on liver tissue and cultured skin fibroblasts revealed a severe glucosidase I deficiency. The residual activity was <3% of that of controls. Glucosidase I activities in cultured skin fibroblasts from both parents were found to be 50% of those of controls. Tissues from the patient subjected to SDS-PAGE followed by immunoblotting revealed strongly decreased amounts of glucosidase I protein in the homogenate of the liver, and a less-severe decrease in cultured skin fibroblasts. Molecular studies showed that the patient was a compound heterozygote for two missense mutations in the glucosidase I gene: (1) one allele harbored a G-->C transition at nucleotide (nt) 1587, resulting in the substitution of Arg at position 486 by Thr (R486T), and (2) on the other allele a T-->C transition at nt 2085 resulted in the substitution of Phe at position 652 by Leu (F652L). The mother was heterozygous for the G-->C transition, whereas the father was heterozygous for the T-->C transition. These base changes were not seen in 100 control DNA samples. A causal relationship between the alpha-glucosidase I deficiency and the disease is postulated.     

de novo转录组学分析华南地区入侵植物五爪金龙代谢特征

为探究华南地区严重入侵植物五爪金龙(Ipomoea cairica)生物入侵的分子机制,对五爪金龙及其近缘种七爪金龙(I. digitata)和裂叶牵牛(I. nil)进行de novo转录组测序和组装,得到56551条unigenes,其中56522条得到注释,7815条GO注释,15615条COG注释,180201条KEGG数据库注释。转录组分析结果表明,五爪金龙氮代谢通路关键酶基因的表达高于对照。次生代谢关键酶(PAL、4CL、CAD、查耳酮合酶、苯基丙乙烯酮异构酶、槲皮黄3-O-甲基转移酶等)基因在五爪金龙与七爪金龙及裂叶牵牛中均得到协同性的差异表达,而这些代谢通路指导的产物合成对五爪金龙的抗逆境能力、生长、化感作用等均起关键作用。关键基因的RT-qPCR验证结果与转录组结果具有一致性。因此,这从分子生物学层面上对解释五爪金龙在华南地区的入侵机制提供了新的证据。     

Designing cross-linked lipase aggregates for optimum performance as biocatalysts

Cross-linked enzyme aggregates (CLEAs) are prepared by precipitation of an enzyme and then chemical cross-linking the precipitate. Three CLEAs of lipase with glutaraldehyde concentrations of 10 mM (CLEA A), 40 mM (CLEA B) and 60 mM (CLEA C) were prepared. Studies show that there is a trade-off between thermal stability vs transesterification/hydrolysis rate vs enantioselectivity. The initial rates for transesterification of β-citronellol for the uncross-linked enzyme and CLEAs A, B and C were 243, 167, 102 and 40 µmol mg^?1 h^?1, respectively. Their thermal stabilities in aqueous media, as reflected by their half-life values at 55°C, were 6, 9, 13 and 16 h, respectively. The enantioselectivity, E values (for kinetic resolution of β-citronellol by transesterification) were 19, 74, 11 and 6, respectively. These results show that CLEA C was the most thermostable; the uncross-linked enzyme was best at obtaining the highest transesterification rate; and CLEA A was best suited for the enantioselective synthesis. Scanning electron microscopy (SEM) showed that the morphology of CLEA was dependent upon the extent of cross-linking.     

The Ultrastructure of the Fungus Trichoderma viride and Investigation of its Growth on Cellulose

Changes in the ultrastructure of Trichoderma viride during growth in shake cultures on cellobiose and cellulose fibres were examined. Electron micrographs of thin sections of germinating conidia, septate hyphae with ascomycete pores and other cell organelles are presented. Extensive autolysis of hyphae was observed after growth for 20 h on cellobiose. The fungus grew in the lumina and within the walls of cellulose fibres. The hyphae followed the directions of the laminar structure but did not grow across them. The observations indicated that the hyphae penetrated the fibres by causing cracks and by dissolving enzymatically the cellulose.     

Photoluminescence of Alq3‐ and Tb‐activated aluminium–tris(8‐hydroxyquinoline) complex for blue chip‐excited OLEDs

The tris(8‐hydroxyquinoline)–aluminium complex is the most important and widely studied as electron transporting and green light emitting material. Alq₃ and Tb_xAl_(1‐x)q₃ have been synthesized (where x = 0.1, 0.3, 0.5, 0.7 and 0.9) and blended films of Alq₃ and Tb_xAl_(1‐x)q₃ with PMMA and PS at different percentage weight (wt%) concentrations (e.g., 0.1, 1, 5, 10, 25 and 50 wt%) have been prepared. The synthesized materials and their blended thin films have been characterized by a photoluminescence (PL) technique; the synthesis and PL characterization are reported in this paper. The synthesized metal complex shows bright emission of green light with blue light excitation (440 nm) and the prepared Tb_xAl_(1‐x)q₃ phosphor may be applicable in blue chip‐excited OLEDs for the newly developed wallpaper lighting technology. Copyright © 2012 John Wiley & Sons, Ltd.