Mutagenic effect of ultraviolet radiation on the non-acid-fast strain ofMycobacterium phlei

The mutability of the PN strain ofMycobacterium phlei was examined after induction of auxotrophic mutants and of STM and VM-resistant mutants, by UV irradiation. A total of 30 auxotrophic mutants were isolated, most of them amino acid-dependent five purine-dependent, and one uracil-dependent. To induce the mutants higher UV doses had to be used so that the survival of cells in the original suspension would not exceed a few per cent. For further genetic work use can be made of 8 auxotrophic mutants (PN try^?ura^?, PN arg^?ura^?, PN ileu^?val^?, PN ileu^?, PN leu^?, PN lys^?, PN lys^?-VM^r, PN val^?), these showing a low frequency of spontaneous reversions. No spontaneous auxotrophic mutants have been found. The frequency of STM and VM-resistant mutants is increased upon UV irradiation, a post-irradiation incubation in a liquid medium without the drug being essential for their phenotypic expression. The highest increase of the number of these mutants is attained after 48 h of post-irradiation incubation and it has been found that, within a certain experimental scatter, the same frequency increase is found on using a complete or a minimal liquid medium. The frequency of spontaneous STM-resistant mutants lies within 5.8×10^?6–8.8×10^?6, of those VM-resistant between 3.1×10^?5 and 4.1×10^?5. The highest frequency of induced STM-resistant mutants lies between 3.0×10^?5 and 9.3×10^?5 and of VM-resistant mutants between 1.1×10^?4 and 2.2×10^?4     

Purification and properties of a peptic heme peptide from cytochrome c1.

Highly purified mouse liver plasma membranes have been used to define the properties of an NADH dehydrogenase activity associated with plasma membrane. The NADH indophenol reductase activity is two-fold stimulated at 5 × 10^?8 M glucagon and the stimulation is inhibited by atebrin. Corresponding activity in endoplasmic reticulum is not stimulated by glucagon. The NADH indophenol reductase is 90% inhibited by insulin at 7 × 10^?11M and shows a return to the original activity at higher insulin concentrations. NADH dehydrogenase activity in endoplasmic reticulum is inhibited up to 50% by insulin at a similar concentration. Triiodothyronine at 10^?7M also inhibits the plasma membrane dehydrogenase whereas thyroxine has little effect. The response of this dehydrogenase to hormones suggests a role in regulation of cellular function.     

Increased synthesis of α‐tocopherol,paramylon and tyrosine by Euglena gracilis under conditions of high biomass production

Aims: To analyse the production of different metabolites by dark‐grown Euglena gracilis under conditions found to render high cell growth. Methods and Results: The combination of glutamate (5 g l^?1), malate (2 g l^?1) and ethanol (10 ml l^?1) (GM + EtOH); glutamate (7·15 g l^?1) and ethanol (10 ml l^?1); or malate (8·16 g l^?1), glucose (10·6 g l^?1) and NH₄Cl (1·8 g l^?1) as carbon and nitrogen sources, promoted an increase of 5·6, 3·7 and 2·6‐fold, respectively, in biomass concentration in comparison with glutamate and malate (GM). In turn, the production of α‐tocopherol after 120 h identified by LC‐MS was 3·7 ± 0·2, 2·4 ± 0·1 and 2 ± 0·1 mg [g dry weight (DW)]^?1, respectively, while in the control medium (GM) it was 0·72 ± 0·1 mg (g DW)^?1. For paramylon synthesis, the addition of EtOH or glucose induced a higher production. Amino acids were assayed by RP‐HPLC; Tyr a tocopherol precursor and Ala an amino acid with antioxidant activity were the amino acids synthesized at higher concentration. Conclusions: Dark‐grown E. gracilis Z is a suitable source for the generation of the biotechnologically relevant metabolites tyrosine, α‐tocopherol and paramylon. Significance and Impact of the Study: By combining different carbon and nitrogen sources and inducing a tolerable stress to the cell by adding ethanol, it was possible to increase the production of biomass, paramylon, α‐tocopherol and some amino acids. The concentrations of α‐tocopherol achieved in this study are higher than others reported previously for Euglena, plant and algal systems. This work helps to understand the effect of different carbon sources on the synthesis of bio‐molecules by E. gracilis and can be used as a basis for future works to improve the production of different metabolites of biotechnological importance by this organism.     

Characterization and Regulatory Properties of Glucose-6-Phosphate Dehydrogenase from Black Gram (Phaseolus mungo)

Glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49) was partially purified by fractionation with ammonium sulfate and phosphocellulose chromatography. The K_m value for glucose-6-phosphate is 1.6 × 10^?4 and 6.3 × 10^?4M at low (1.0–6.0 × 10^?4M) and high (6.0–30.0 × 10^?4M) concentrations of the substrate, respectively. The K_m value for NADP⁺ is 1.4 × 10^?5M. The enzyme is inhibited by NADPH, 5-phosphoribosyl-1-pyrophosphate, and ATP, and it is activated by Mg²⁺, and Mn²⁺. In the presence of NADPH, the plot of activity vs. NADP⁺ concentration gave a sigmoidal curve. Inhibition of 5-phosphoribosyl-1-pyrophosphate and ATP is reversed by Mg²⁺ or a high pH. It is suggested that black gram glucose-6-phosphate dehydrogenase is a regulatory enzyme of the pentose phosphate pathway.     

Evaluating different carriers of Metarhizium brunneum F52 microsclerotia for control of adult Asian longhorned beetles (Coleoptera: Cerambycidae)

Microsclerotia (MS) of Metarhizium brunneum strain F52 (Hypocreales: Clavicipitaceae) were processed as granules using three carriers: kaolin clay, microcrystalline cellulose (MCC), or diatomaceous earth (DE). In a series of experiments aimed at comparing viable conidial production, each MS carrier type was hydrated using a decreasing range of water activities on glycerol or polyethylene glycol (PEG200)-amended media. Conidial density and percent germination of conidia declined significantly as water activity was lowered. All three carrier types produced >2–6?×?10⁹ viable conidia/gram at higher water activities (а_w >0.987) but were still capable of producing 2.9?×?10⁷?±?1.6?×?10⁶ conidia/g (30% glycerol а_w?=?0.883) and 1.9?×?10⁷?±?6.5?×?10⁶ conidia/g (30% PEG200 а_w?=?0.924) at the lowest tested water activities. MS carrier types were sprayed onto 0.4?m length logs with or without hydromulch formulation. The median survival times of Asian longhorned beetles, Anoplophora glabripennis (Coleoptera: Cerambycidae), exposed by climbing double the length of the logs of hydromulch-treated bark, were 16.5–20.5?d while beetles exposed to logs without hydromulch formulation had median survival times of 22–25.5?d. Overall, experiments showed that there were few biologically significant differences between the MS carrier types.     

Behavioural responses of sablefish, Anoplopoma fimbria, to bait odour

A behavioural bioassay was used to determine the response threshold to squid extract of sablefish, Anoplopoma fimbria, held at three different feeding regimens. Sablefish responded to the odour of bait by changing swimming activity and turning behaviour. The response threshold to bait odour was influenced by both the amount of food eaten and the duration of food deprivation. The total concentration of amino acids in the bait extract was assumed to determine the response threshold as chemical fractionation studies have shown that this class of compounds is essential for the stimulatory capacities of food extracts. When fed to satiation (9.4% wet body weight) and tested after one day of food deprivation, the mean response threshold to total dissolved free amino acids was 4.4 × 10^?8m (range=7.6 × 10^?8 to 3.6 × 10^?8m ). When fed at 1.6-2.3% wet body weight, the threshold sensitivity had increased to a mean value of 1.8 × 10^?10m (range=8.4 × 10^?10 to 7.0 × 10^?11m ) after one day of food deprivation; after four days of deprivation, the sensitivity had increased even further to a mean value of 1.4 × 10^?11m (range=1.6 × 10^?10 to 1.4 × 10^?12m ). It was also apparent that the intensity of behavioural responses to the bait odour increased with both stimulus concentration and duration of food deprivation. These results suggest that sablefish intensify their search for prey under increased feeding motivation. The active space of a bait source was estimated from the threshold values obtained. Depending on state of food deprivation, rate of chemical release from the bait and the current velocity, maximum lengths of active space within which sablefish would exhibit food searching responses vary from 10 m to several km. Stock assessment based on catch data from baited gear will need techniques that take into account those factors influencing active space for food searching.     

Differential in vitro inhibition of polyphenoloxidase from a wild edible mushroom Lactarius salmonicolor

The polyphenol oxidase (LsPPO) from a wild edible mushroom Lactarius salmonicolor was purified using a Sepharose 4B-L-tyrosine-p-amino benzoic acid affinity column. At the optimum pH and temperature, the K_M and V_Max values of LsPPO towards catechol, 4-methylcatechol and pyrogallol were determined as 0.025 M & 0.748 EU/mL, 1.809 × 10^{? 3} M & 0.723 EU/mL and 9.465 × 10^{? 3} M & 0.722 EU/mL, respectively.

Optimum pH and temperature values of LsPPO for the three substrates above ranged between the pH 4.5–11.0 and 5–50°C. Enzyme activity decreased due to heat denaturation with increasing temperature. Effects of a variety of classical PPO inhibitors were investigated opon the activity of LsPPO using catechol as the substrate. IC₅₀ values for glutathione, p-aminobenzenesulfonamide, L-cysteine, L-tyrosine, oxalic acid, β-mercaptoethanol and syringic acid were determined as 9.1 × 10^{? 4}, 2.3 × 10^{? 4} M, 1.5 × 10^{? 4} M, 3.8 × 10^{? 7} M, 1.2 × 10^{? 4} M, 4.9 × 10^{? 4} M, and 4 × 10^{? 4} M respectively. Thus L-tyrosine was by far the most effective inhibitor. Interestingly, sulfosalicylic acid behaved as an activator of LsPPO in this study.     

Breeding and characterization of amino acid-analogue-resistant mutants of Arthrospira platensis

To obtain amino acid-analogue-resistant mutants the wild strain A9 of Arthrospira platensis was mutated by ethylmethane sulfonate (EMS). Mutagenic effects of strain A9 by EMS were studied. The experimental results indicated that the survival rate curve of strain A9 took a typical “exponential shape” with lethal dosage of EMS being 1 %. The survival of A9 strain was 13.2 % when treated with 0.4 % of EMS, and the resistant mutation rates to two amino acid analogues, ρ-fluorophenylalanine (FPA) and L-canavanine sulphate (CS), were greatly increased with the highest rates being at 4.9 × 10^?4 and 3.24 × 10^?4, respectively. By repeated screening, two stable mutants resistant to amino acid analogues, A9f resistant to FPA and A9c resistant to CS, were obtained. Resistances of the two mutants to corresponding amino acid-analogues were both significantly increased. Compared with their parent strain A9, A9f appeared larger than A9 performance in filament diameter, spiral diameter, spiral pitch, filament length and spiral number, and A9c showed much longer length and spiral pitch than those of the initial strain. Analysis results on amino acids compositions and contents showed that both two mutants accumulated quite higher concentration of amino acids in cells. The two mutants might be excellent high amino acids producing strain. By this means two useful mutants with stable genetic makers for further genetic study of A. platensis were obtained, which laid a good foundation for further study on the transformation of A. platensis.     

The Influence of pH on the Cadmium-Repressed Growth of the Alga Coelastrum proboscideum

The inhibition of growth by different concentrations of CdCl₂ in the range 4,5 × 10^?7 to 5.6 × 10^?7M was studied in the green alga Coelastrum proboscideum Bohlin in inorganic media at pH 4.3, 5.3 and 6.2. The factorial destgn of the experiments was evaluated as an analysis of 2² factors. Below pH 4.0 and above pH 6.5 growth was depressed without adding Cd. Cd concentrations exceeding 5.6 × 10^?8M reduced algal growth significantly with a 50% inhibition at 5.6 × 10^?7M Cd. The Cd concentration of 5.6 × 10^?7M was less toxic at pH 6.2 than at pH 5.3 and 4.3, thus revealing a negative interaction between protons and Cd.     

Carbon dynamics in an ultra-oligotrophic epishelf lake (Beaver Lake, Antarctica) in summer

1. Microbial plankton dynamics in an ultra‐oligotrophic epishelf lake (Beaver Lake, Antarctica) were investigated over an austral summer (December 2002 to January 2003). The aim was to characterise carbon cycling in an environmentally extreme lake. 2. The lake had an unusual temperature profile with peak temperatures of 1.3–1.9 °C between 20 and 25 m. Photosynthetically active radiation penetrated to the lake bottom (110 m) on occasions. The ice cover underwent marked thinning and melting during the study period. 3. Chlorophyll a concentrations were consistently low, usually below 1 μg L^?1, with highest concentrations close to the lake bottom, where the photosynthetic elements showed strong autofluorescence. Mean photosynthetic nanoflagellates ranged between 34.9 × 10⁴ L^?1 ± 33.5 (23rd December) and 130.9 × 10⁴ L^?1 ± 112.3 (4th December). Highest photosynthetic activity was usually recorded below 25 m. Rates of carbon fixation varied between 0.089 μg C L^?1 h^?1 ± 0.002 and 0.579 μg C L^?1 h^?1 ± 0.156. Primary production was limited by low temperature and orthophosphate availability. 4. Mean bacterial concentration throughout the water column ranged between 9.3 × 10⁷ L^?1 ± 1.2 (23rd December) and 14.0 × 10⁷ L^?1 ± 1.8 (28th January). Bacterial production was low, less than 10% of primary production and ranged between 2.1 ng C L^?1 h^?1 ± 0.8 and 12 ng C L^?1 h^?1 ± 0.9. Highest rates coincided with times of highest primary production. On occasion dissolved organic carbon (DOC) concentrations dropped to 20 μg L^?1, probably below accurate limits of detection, suggesting that carbon substratum and phosphorus may have limited bacterial growth. 5. Heterotrophic nanoflagellates varied significantly over the summer from a mean of 26.6 × 10⁴ L^?1 ± 14.2 (23rd December) to 133.8 × 10⁴ L^?1 ± 33.5 (14th December). They imposed a significant grazing impact on the bacterioplankton, removing in excess of 100% of bacterial production in December. 6. The total organic carbon pool [DOC and particulate organic carbon (POC)] was below 600 μg L^?1. The ratio of DOC : POC ranged between 0.44 : 1 and 2.8 : 1 in the upper 40 m of the water column, and 1.8 : 1 and 3.7 : 1 in the lower waters. The microbial plankton contributed 1–29% of POC, thus detrital POC made up the largest fraction of the POC pool. 7. Beaver Lake is an extreme lacustrine ecosystem where heterotrophic processes occasionally appear to be carbon limited. Significant summer ice‐melt, not seen in a previous opportunistic sampling, may be having an impact on the carbon cycle.     

Three‐dimensional (3‐D) fluorescence spectroscopy analysis of the fluorescent dissolved organic matter released by the marine toxic dinoflagellate Alexandrium catenella exposed to metal stress by zinc or lead

We investigated the effects of zinc or lead on growth and on exudation of fluorescent dissolved organic matter (FDOM) by the marine toxic dinoflagellate Alexandrium catenella (Whedon & Kofoid) Balech. The species was exposed to increasing free zinc (1.34 × 10^?7 M–3.98 × 10^?6 M) or lead (5.13 × 10^?9 M–1.82 × 10^?7 M) concentra‐tions. Low metal levels ([Zn²⁺] = 1.34 × 10^?7 M; [Pb²⁺] = 5.13 × 10^?9 M) had no effect on cell growth. Toxic effects were observed from higher metal contamination ([Zn²⁺] = 3.98 × 10^?6 M; [Pb²⁺] = 6.54 × 10^?8 M), as a conversion of vegetative cells into cysts. Analysis of the released FDOM by three‐dimensional (3‐D) fluorescence spectroscopy was achieved, using the parallel factor analysis (PARAFAC). The PARAFAC modeling revealed four components associated with two contributions: one related to the biological activity; the other linked to the organic matter decomposition in the culture medium. The C1 component combined a tryptophan peak and characteristics of humic substances, whereas the C2 component was considered as a tryptophan protein fluorophore. The two others C3 and C4 components were associated with marine organic matter production. Relea‐sed fluorescent substances were induced by low ([Zn²⁺]= 1.34 × 10^?7 M; [Pb²⁺] = 5.13 × 10^?9 M) and moderate ([Zn²⁺] = 6.21 × 10^?7 M; [Pb²⁺] = 2.64× 10^?9 M) metal concentrations, suggesting the activation of cellular mechanisms in response to metal stress, to exudate FDOM that could complex metal cations and reduce their toxicity toward A. catenella cells.     

Purification and properties of DL-lactate dehydrogenase from Leuconostoc mesenteroides

A dl-lactate dehydrogenase from the bacterium, Leuconostoc mesenteroides, has been purified and characterized with respect to amino acid composition, molecular weight, and kinetic properties. The turnover number of the enzyme was 1.7 × 10⁵ moles DPNH/mole enzyme/min for the most active of three preparations. On the basis of a sedimentation constant of 3.52 S and a diffusion constant of 5.0 × 10^?7 cm²/ ml, the molecular weight of the enzyme was determined to be approximately 64,000. Similar values were derived from sedimentation equilibrium data. The enzyme exhibits typical Michaelis-Menten kinetics except when lactate is the variable substrate. In this case, double reciprocal plots of activity versus substrate concentration are curved upward, suggesting that lactate either activates or stabilizes a more active form of the enzyme.     

Characterization of the Galactose-Specific Binding Activity of a Purified Soluble Entamoeba histolytica Adherence Lectin

ABSTRACT. We studied galactose (Gal)-specific binding of the soluble purified 260-kDa Entamoeba histolytica adherence protein to glycosylation deficient Chinese hamster ovary (CHO) cell mutants. Our goal was to further define the lectin's functional activity and carbohydrate receptor specificity. The adherence protein was purified by acid elution from an immunoaffnity column; however, exposure of the surface membrane lectin on viable trophozoites to identical acid pH conditions had no effect on carbohydrate binding activity. Saturable Gal-specific binding of soluble lectin to parental CHO cells was demonstrated at 4°C by radioimmunoassay; the dissociation coefficient (Kd was 2.39 × 10^?8 M^?1 with 5.97 × 10⁴ lectin receptors present per CHO cell. Gal-specific binding of lectin to Lec2 CHO cell mutants, which have increased N- and O-linked terminal Gal residues on cell surface carbohydrates, was increased due to an enhanced number of receptors (2.41 × 10⁵/cell) rather than a significantly reduced dissociation constant (4.93 × 10^?8 M^?1). At 4°C, there was no measurable Gal-specific binding of the adherence protein to the Lec and IdlD.Lecl CHO mutants, which contain surface carbohydrates deficient in terminal Gal residues. Binding of lectin (20 μg/ml) to CHO cells was equivalent at 4°C and 37°C and unaltered by adding the microfilament inhibitor, Cytochalasin D (10 μg/ml). Gal-specific binding of the lectin at 4°C was calcium independent and reduced by 81% following adsorption of only 0.2% of the lectin to CHO cells. In summary, these findings indicate that the purified E. histolytica adherence lectin demonstrates saturable Gal-specific binding to 1–6 branched-N-linked and not O-linked galactose terminal cell surface carbohydrates; however, apparently only a small percentage of purified amebic lectin molecules actually possess galactose binding activity.     

Selective determination of human immunoglobulin G by flow‐injection chemiluminescence

A novel flow‐injection chemiluminescence method was developed for the selective determination of human immunoglobulin G (IgG) in the presence of thiomersal by changing the flow rates of peristaltic pump. The study was based on the independence and additivity of the CL signals of human IgG and thiomersal in the galangin–potassium permanganate–polyphosphoric acid system. In meantime, two equations relating to the concentrations of mixing solutions of human IgG and thiomersal vs the CL intensity were established and solved, on the basis of which the content of thiomersal included in samples was simultaneously determined too. The enhanced CL intensity was in proportion to concerntrations in the range 8.0 × 10^?7 to 8.0 × 10^?5 g/mL for human IgG and 1.0 × 10^?7 to 2.0 × 10^?6 g/mL for thiomersal with the detection limits of 5.0 × 10^?7 g/mL for human IgG and 6.0 × 10^?8 g/mL for thiomersal, respectively. The relative standard deviation for 1.0 × 10^?5 g/mL human IgG was 0.8% and for 2.0 × 10^?7 g/mL thiomersal it was 2.0% (n = 10). The proposed method was applied to determine three synthetic samples with recoveries of 91.5–109.5%. In addition, the possible chemiluminescence mechanisms are discussed as well. Copyright © 2010 John Wiley & Sons, Ltd.     

Glucose-6-phosphate Dehydrogenase and 6-Phosphogluconate Dehydrogenase of Candida tropicalis Partial Purification and Some Properties

Glucose-6-phosphate (G6P) dehydrogenase and 6-phosphogluconate (6PG) dehydrogenase were partially purified about 53-fold and 47-fold, respectively, from the cell-free extract of glucose-grown Candida tropicalis by means of ammonium sulfate fractionation and DEAE-cellulose column chromatography. AMP acted as the competitive inhibitor against G6P and NADP in the G6P dehydrogenase reaction. This inhibition was remarkable at low concentrations of NADP, increasing the sigmoidicity of the NADP-saturation curve. On the other hand, 6PG dehydrogenase was not affected by AMP. Fructose-1,6-bisphosphate (FDP) and phosphoenolpyruvate (PEP) inhibited slightly G6P dehydrogenase. 6PG dehydrogenase was also weakly inhibited by FDP. Apparent Km values of G6P dehydrogenase were calculated as 1.8 × 10^?4 m for G6P and 3.1 × 10^?5 m for NADP. Those of 6PG dehydrogenase were 9.4 × 10^?5 m for 6PG and 2.8 × 10^?5 m for NADP.     

Copper nanocluster‐enhanced luminol chemiluminescence for high‐selectivity sensing of tryptophan and phenylalanine

A remarkable method for the highly sensitive detection of phenylalanine and tryptophan based on a chemiluminescence (CL) assay was reported. It was found that fluorescent copper nanoclusters capped with cysteine (Cys‐CuNCs) strongly enhance the weak CL signal resulting from the reaction between luminol and H₂O₂. Of the amino acids tested, phenylalanine and tryptophan could enhance the above CL system sensitively. Under optimum conditions, this method was satisfactorily described by a linear calibration curve over a range of 1.0 × 10^?6 to 2.7 × 10^?5 M for phenylalanine and 1.0 × 10^?7 to 3.0 × 10^?5 M for tryptophan, respectively. The effect of various parameters such as Cys‐CuNC concentration, H₂O₂ concentration and pH on the intensity of the CL system were also studied. The main experimental advantage of the proposed method was its selectivity for two amino acids compared with others. To evaluate the applicability of the method to the analysis of a real biological sample it was used to determine tryptophan and phenylalanine in human serum and remarkable results were obtained.     

The fluorescence enhancement of quercetin–nucleic acid system and the analytical application

Nucleic acid can greatly enhance the fluorescence intensity of quercetin in HMTA‐HCl (pH 5.5) buffer. The enhanced intensity is in proportion to the concentration of nucleic acids in the range 5.0 × 10^?9 to 1.0 × 10^?6 g/mL for fsDNA, 5.0 × 10^?9 to 7.0 × 10^?7 g/mL for ctDNA and 5.0 × 10^?9 to 1.0 × 10^?6 g/mL for yRNA, and their detection limits (S/N = 3) are 3.5 × 10^?9, 7.8 × 10^?10 and 2.6 × 10^?9 g/mL, respectively. In comparison with most reported fluorescent probes for the determination of nucleic acids, the proposed probe has higher sensitivity and lower toxicity. The interaction investigation indicates that quercetin binds with double‐strand DNA in groove binding mode, resulting in fluorescence enhancement of this system. Copyright © 2009 John Wiley & Sons, Ltd.     

α-Glucosidase Inhibitor from the Seeds of Balsam Pear (Momordica charantia) and the Fruit Bodies of Grifola frondosa

α-Glucosidase inhibitory activities were found in aqueous methanol extracts of the seeds of Momordica charantia and the fruit bodies of Grifola frondosa. An active principle against the enzyme prepared from rat small intestine acetone powders was isolated and characterized. The structure of the isolated compound was identified as D-(+)-trehalose by FDMS, ¹H-, ¹³C-NMR, and [α]_D measurements. The inhibitory activity of trehalose was compared with 1-deoxynojirimycin. Trehalose showed 45% inhibitory activity at the concentration of 2×10^?3 M, but 1-deoxynojirimycin had 52% inhibitory activity at 1×10^?7 M.     

Thulium Ion Promotes Apoptosis of Primary Mouse Bone Marrow Stromal Cells

Bone is one of the main target organs for the lanthanides (Ln). Biodistribution studies of Tm-based compounds in vivo showed that bone had significant uptake. But the effect of Tm³⁺ on primary mouse bone marrow stromal cells (BMSCs) has not been reported. So we investigated the effect and underlying mechanisms of Tm³⁺ on BMSCs. Cell viability, cell apoptosis, reactive oxygen species (ROS) level, lactate dehydrogenase (LDH) activity and mitochondrial membrane potential (MMP) were studied. The results indicated that Tm³⁺ increased the viability of BMSCs at concentrations of 1?×?10^?7, 1?×?10^?6, 1?×?10^?5, and 1?×?10^?4 mol/L in a dose-dependent manner, turned to decrease the viability of BMSCs at the highest concentration of 1?×?10^?3 mol/L for 24, 48, and 72 h. Tm³⁺ at 1?×?10^?3 mol/L promoted apoptosis of BMSCs, increased the ROS and LDH levels, and decreased MMP in BMSCs. Taken together, we demonstrated that Tm³⁺ at 1?×?10^?3 mol/L might induce cellular apoptosis through mitochondrial pathway. These results may be helpful for more rational application of Tm-based compounds in the future.     

Quorum sensing activity in Ophiostoma ulmi: effects of fusel oils and branched chain amino acids on yeast-mycelial dimorphism

Aims: For Ophiostoma (Ceratocystis) ulmi, the ability to undergo morphological change is a crucial factor for its virulence. To gain an understanding of quorum‐sensing activity in O. ulmi as it relates to yeast‐mycelium dimorphism control, this study examines the effects of branched‐chain amino acids as well as their fusel alcohols and fusel acids as quorum sensing molecules. Methods and Results: In a defined medium containing glucose, proline and salts, O. ulmi grew as yeasts when the culture was inoculated with a high density of spores (2 × 10⁷ CFU ml^?1) and as mycelia when inoculated with a low spore density (4 × 10⁵ CFU ml^?1). The cultures displaying yeast morphology secreted a quorum‐sensing factor that shifted the morphology from mycelia to yeast. This quorum‐sensing molecule was lipophilic and extractable by organic solvents from the spent medium. Using GC/MS analysis, it was determined that the major compound in the extract was 2‐methyl‐1‐butanol. A similar effect was observed when the branched‐chain amino acids (fusel alcohol precursors) were used as the nitrogen source. E, E‐farnesol had no effect on the morphology of O. ulmi. Conclusions: Addition of the branched‐chain amino acids or one of the compounds detected in the spent medium, 2‐methyl‐1‐butanol or 4‐hydroxyphenylacetic acid, or methylvaleric acid, decreased germ tube formation by more than 50%, thus demonstrating a quorum sensing molecule behaviour in O. ulmi cultures. Significance and impact of the study: This study presents advances in the investigation of dimorphism in O. ulmi, complementing the existing scientific basis, for studying, understanding and controlling this phenomenon.