[Effect of ZnSO4 on L-lactic acid production by Rhizopus oryzae].

OBJECTIVE: To improve the yield and quality of L-lactic acid by Rhizopus oryzae, we aim to understand the relationship between inorganic salts utilization and the L-lactic acid metabolism of the strain RLC41-6, through systematic analysis of the effects of zinc ion concentration on the production of L-lactic acid and the Lactic Dehydrogenase (LDH) activity. METHODS: Rhizopus oryzae was cultured at 36 degrees C for 36h with different quantity of ZnSO4 in fermentation medium. The fermentation products were monitored by reversed-phase high performance liquid chromatography (RP-HPLC), LDH isoenzyme composition in the cell was analysed by non-denatured polyacrylamide gel electrophoresis (PAGE). RESULTS: Our results showed that the concentration of ZnSO4 in medium could modulate the expression of LDH isoenzyme except LDH1, especially stimulated the expression of LDH4 and LDH5. When initial concentration of ZnSO4 is above 0.02%, the LDH4 and LDH5 reached the highest level. However, the activity of LDH was inhibited by higher concentration zinc ion in extracellular environment. When ZnSO4 concentration is 0.02%, LDH activity reaches its maximum 200U/mL, the HPLC assay showed only L-lactic in the fermentation products (137 g/L), while the conversion rate of glucose to lactic acid is 91%. CONCLUSION: Zinc ion can regulate the metabolic processes of Rhizopus oryzae and modulate the types of the final fermentation products. An optimal concentration of ZnSO4 can not only facilitate the LDH expression but also prevent pyruvate from transformation into the malic acid and fumaric acid during the metabolism process, thereby enhance the metabolism of glucose to lactic acid of Rhizopus oryzae.

Assessing the impacts of fine sediment removal on endogenous pollution release and microbial community structure in the shallow lakes.

Resuspension is a crucial process for releasing endogenous pollution from shallow lakes into the overlying water. Fine particle sediment, which has a higher contamination risk and longer residence time, is the primary target for controlling endogenous pollution. To this end, a study coupling aqueous biogeochemistry, electrochemistry, and DNA sequencing was conducted to investigate the remediation effect and microbial mechanism of sediment elution in shallow eutrophic water. The results indicated that sediment elution can effectively remove some fine particles in situ. Furthermore, sediment elution can inhibit the release of ammonium nitrogen and total dissolved phosphorous into the overlying water from sediment resuspension in the early stage, resulting in reductions of 41.44 %-50.45 % and 67.81 %-72.41 %, respectively. Additionally, sediment elution greatly decreased the concentration of nitrogen and phosphorus pollutants in pore water. The microbial community structure was also substantially altered, with an increase in the relative abundance of aerobic and facultative aerobic microorganisms. Redundancy analysis, PICRUSt function prediction, and the correlation analysis revealed that loss on ignition was the primary factor responsible for driving changes in microbial community structure and function in sediment. Overall, the findings provide novel insights into treating endogenous pollution in shallow eutrophication water.

Reduction and removal of aqueous Cr(VI) by glow discharge plasma at the gas-solution interface.

Aqueous chromium(VI) reduction and removal induced by glow discharge taking place at the gas-solution interface in an argon atmosphere was studied. The effect of initial pH and hydroxyl radical scavenger (ethanol) on the reduction efficiency was examined. High reduction efficiency was obtained when initial pH ≤ 2.0 or ≥ 8.0. In particular, addition of ethanol into the solution substantially increased the reduction efficiency and facilitated chromium removal from the solution in the form of sediment after discharge. The optimum pH values for Cr(VI) removal were within 6.0-7.0. Fourier transform-infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analysis confirmed that the main constituent of the sediment is chromium hydroxide.

Toxicity of carbon nanotubes to p21 and hus1 gene deficient mammalian cells.

Carbon nanotubes including single wall and multi wall carbon nanotubes (SWNT and MWNT) are attractive nanomaterials with great promise in industrial and medical applications. However, little is known about the role of p21 and hus1 gene in the toxic response of SWNT and MWNT to mammalian cells. The aim of this study is to investigate the role of the p21 and hus1 genes in the toxicity of carbon nanotubes. Comparison of micronucleus fraction between the wild type and p21 -/- , hus1 +/+ mouse embryo fibroblast (MEF) cells was performed experimentally. Our results show that the yield of the micronucleus ratio in p21 gene knock-out MEF cells is lower than that in the wild type counterpart, indicating that p21 may play as anti-apoptosis factor during the signal transduction of DNA damage caused by carbon nanotubes in mammalian cells.

Genotoxicity/mutagenicity of formaldehyde revealed by the Arabidopsis thaliana plants transgenic for homologous recombination substrates.

Formaldehyde (FA) is a major industrial chemical and has been extensively used in the manufacture of synthetic resins and chemicals. The use of FA-containing industrial materials in daily life exposes human to FA extensively. Numerous studies indicate that FA is genotoxic, and can induce various genotoxic effects in vitro and in vivo. The primary DNA lesions induced by FA are DNA-protein crosslinks (DPCs). Recently, it has been reported that the homologous recombination (HR) mechanism is involved in the repair of DPCs, suggesting the homologous recombination could be a potential indicator for the genotoxicity/mutagenicity of FA. However, it has not yet been reported that organisms harboring recombination substrates are used for the detection of genotoxic/mutagenic effects of FA. In this present study, an Arabidopsis thaliana-line transgenic for GUS recombination substrates was used to study the genotoxicity/mutagenicity of FA, and the results showed that FA-exposure significantly increased the induction of HR in growing plants, but not in dormant seeds. We also observed an early up-regulation of expression of HR-related gene, AtRAD54, after FA-exposure. Moreover, the pretreatment with glutathione (GSH) suppressed drastically the induction of HR by FA-exposure.

Nickel-induced apoptosis and relevant signal transduction pathways in Caenorhabditis elegans.

Many investigations have shown that nickel exposure can induce micronuclei generation, inhibit DNA repair and induce cell apoptosis, both in cells and tissues. However, there is a lack of appropriate in vivo animal models to study the underlying mechanisms of nickel-induced apoptosis. The model organism, Caenorhabditis elegans, has been shown to be a good model for investigating many biological processes. In the present study, we detected 0.01 mM nickel induced significantly germline cell apoptosis after treatment for 12 hours, which demonstrated that C. elegans could be a mammalian in vivo substitute model to study the mechanisms of apoptosis. Then gene knockout C. elegans strains were utilized to investigate the relationship between nickel-induced apoptosis and relevant signal pathways, which were involved in DNA damage and repair, apoptosis regulation and damage signal transduction. The results presented here demonstrated that nickel-induced apoptosis was independent of the DNA damage response gene, such as hus-1, p53/cep-1 and egl-1. The loss-of-function of the genes that related to Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinases (MAPK) signaling cascades suppressed nickel-induced germline apoptosis, while ERK signaling cascades have no effects on the nickel-induced germline apoptosis.

A unique technology to transform inorganic nanorods into nano-networks.

An inorganic nano-network of attapulgite is formed from rigid nanorods using ion beam bombardment. The structure of the nano-networks depends on the ion beam fluence for the same ion energy. Scanning electron microscopy reveals that ion beam bombardment improves the dispersion of the attapulgite particles and the change in the shape of the rod-shaped attapulgite particles stems from the thermal stress induced by ion beam bombardment. This phenomenon is more obvious for higher ion fluences. The bent or twisted rod-shaped attapulgite particles cross-link to form a network structure, which is stable in water, and when the ion fluence is increased further, the cross-linked points are permanently sealed. The improved materials are more useful than clava attapulgite particles.

Enhanced nitrogen and phosphorus removal from eutrophic lake water by Ipomoea aquatica with low-energy ion implantation.

Ipomoea aquatica with low-energy N+ ion implantation was used for the removal of both nitrogen and phosphorus from the eutrophic Chaohu Lake, China. The biomass growth, nitrate reductase and peroxidase activities of the implanted I. aquatica were found to be higher than those of I. aquatica without ion implantation. Higher NO3-N and PO4-P removal efficiencies were obtained for the I. aquatica irradiation at 25 keV, 3.9 x 10(16) N+ ions/cm(2) and 20 keV 5.2 x 10(16) N+ ions/cm(2), respectively (p < 0.05). Moreover, the nitrogen and phosphorus contents in the plant biomass with ion implantation were also greater than those of the controls. I. aquatica with ion implantation was directly responsible for 51-68% N removal and 54-71% P removal in the three experiments. The results further confirm that the ion implantation could enhance the growth potential of I. aquatica in real eutrophic water and increase its nutrient removal efficiency. Thus, the low-energy ion implantation for aquatic plants could be considered as an approach for in situ phytoremediation and bioremediation of eutrophic waters.

L(+)-lactic acid production by co-fermentation of glucose and xylose with Rhizopus oryzae obtained by low-energy ion beam irradiation.

Low-energy ion beam irradiation (10-200 keV) has been proved to have a wide range of biological effects in recent years. When Rhizopus oryzae PW352 was irradiated with a 15-keV low-energy ion beam an L(+)-lactic acid high-yield mutant, RQ4015, was obtained. When 150 g/l glucose was used as the sole carbon source, L(+)-lactic acid of RQ4015 reached 121 g/l after 36 h shake-flask cultivation. However, the highest lactic acid concentration 74 g/l was obtained when 100 g/l xylose was present in the medium as the sole carbon source. When mixed xylose (25 g/l) and glucose (75 g/l) were present in a bubble column, L(+)-lactic acid production of RQ4015 reached 83 g. A high mutation rate and a wide mutation spectrum of low-energy ion implantation were observed in the experiment, suggesting that ion implantation can be a highly efficient mutagenic means for microorganism breeding in many commercial applications.

Silicon-induced DNA damage pathway and its modulation by titanium plasma immersion ion implantation.

Micronuclei tests (MNT) using the Chinese Hamster Ovary (CHO) cells and Chinese Hamster Ovary Mutant cells (XRS5) have been conducted to evaluate the biocompatibility of silicon and titanium plasma-implanted silicon. Pure Si induces high MN ratios of the two cell lines and thus has poor biocompatibility. The MN ratio of CHO cells is higher than background by about 44% and the MN ratio of XRS5 cells is even higher by about 180%, suggesting that most of the cellular DNA damages on the Si wafer are DNA double-strand breaks (DSB) and are efficiently repaired by the nonhomologous end-joining (NHEJ) pathway. The surface biocompatibility of Si can be enhanced by Ti plasma immersion ion implantation (PIII). The altered oxidized species on the Ti plasma-implanted surface block cellular DSB repaired by the NHEJ pathway and decrease the MN ratio of XRS5 cells. By increasing the Ti implantation time and consequently the Ti implant fluence, the oxygen binding energy shifts toward a lower energy and the intensity of the Si peaks corresponding to SiO(2) continually diminishes and even disappear. At the same time, the MN ratios of the two cell lines decrease. Our results suggest that the rest of the DNA damages which cannot be repaired by the NHEJ pathway may be blocked because the surface bonding changes from predominantly Si-O on the 10 min Ti-implanted Si to Ti-O on the 120 min Ti-implanted Si. Our results also suggest that the genotoxicity of cell assay such as MNT and DSB is a valid method to investigate biocompatibility.

Antagonistic effects of volatiles generated by Bacillus subtilis on spore germination and hyphal growth of the plant pathogen, Botrytis cinerea.

Botrytis cinerea is one of the most serious post-harvest pathogens of fruits and vegetables. Volatiles generated by Bacillus subtilis JA significantly inhibited both spore germination and elongation of germ tubes in Botrytis cinerea using a two-compartment agar-plate assay. The volatiles caused protoplasm retraction from the hyphal tips to the spores.

Damage effects induced by electrically generated negative air ions in Caenorhabditis elegans.

Electrically generated negative air ions (ENIs) have been widely used to improve indoor air quality. However, the effects of ENIs reported so far were inconsistent due to the variance in test systems, end points detected and the exposure methods. In this study, a simple model organism, the nematode Caenorhabditis elegans, was used as an in vivo system to assess the biological effects of continuous ENIs exposure. The worms were exposed to ENIs in a 10(5) 10(5) ions/cm(3) chamber and their development period, lifespan, brood size and germline cell apoptotisis were examined. The results showed that ENIs decreased the development period, shortened the lifespan, increased the germline cell apoptosis and reduced the brood size, suggesting that persistent ENIs exposure might induce damage in C. elegans. To further scrutinize the mechanisms underlying these damage effects, a reactive oxygen species (ROS) sensitive C. elegans, mev-1(kn1) mutant, and sod-3::gfp transgenic strains were used. The results showed that the persistent ENIs exposure significantly shortened the lifespan of mev-1(kn1) mutant compared to the wild type. Moreover, levels of SOD-3 were increased in an exposure time-dependent manner. Treatment with either DMSO or l-ascorbic acid, effective ROS scavengers, could rescue the upregulation of germline cell apoptosis and SOD-3 level induced by ENIs exposure, indicating that ROS may be involved in ENIs exposure-induced damaging effects.

[Purification and identification of iturin A from Bacillus subtilis JA by electrospray ionization mass spectrometry].

Lipopeptides produced by Bacillus subtilis JA antagonized a broad spectrum of plant fungal pathogens. The purification and identification of the lipopeptide antibiotics plays an important role for further research. Crude lipopeptides were extracted with methanol from the precipitate, which was obtained by adding 6mol/L HCl to the cell-free culture broth and then stored at 4 degrees C overnight. The crude extract was run on reversed-phase HPLC system with a Diamonsil C18 column (250 mm x 4.6 mm, Dikma) to separate the lipopeptides. Two antifungal compounds, which had strong inhibitory activity against various plant fungal pathogens, such as Fusarium graminearum, were purified. The molecular weights of two compounds were determined by electrospray ionization mass spectrometry (ESI/MS). Two compounds, with molecular weights of 1042.4 Da and 1056.5 Da, were homologues differed by a structure of -CH2. ESI collision induced dissociation mass spectrometry analysis was used to sequence the structure of purified compounds. Typical b- and y- type fragments showed that compound 1 (with a molecular weight of 1042.4 Da) had a primary structure of Pro-Asn-Tyr-PAA-Asn-Tyr-Asn-Gln (PAA represented beta-amino acid), which was consistent with lipopeptide iturin A. Compound 2 was a homologue of iturin A.

New insight into intrachromosomal deletions induced by chrysotile in the gpt delta transgenic mutation assay.

BACKGROUND: Genotoxicity is often a prerequisite to the development of malignancy. Considerable evidence has shown that exposure to asbestos fibers results in the generation of chromosomal aberrations and multilocus mutations using various in vitro approaches. However, there is less evidence to demonstrate the contribution of deletions to the mutagenicity of asbestos fibers in vivo. OBJECTIVES: In the present study, we investigated the mutant fractions and the patterns induced by chrysotile fibers in gpt delta transgenic mouse primary embryo fibroblasts (MEFs) and compared the results obtained with hydrogen peroxide (H2O2) in an attempt to illustrate the role of oxyradicals in fiber mutagenesis. RESULTS: Chrysotile fibers induced a dose-dependent increase in mutation yield at the redBA/gam loci in transgenic MEF cells. The number of lambda mutants losing both redBA and gam loci induced by chrysotiles at a dose of 1 microg/cm(2) increased by > 5-fold relative to nontreated controls (p < 0.005). Mutation spectra analyses showed that the ratio of lambda mutants losing the redBA/gam region induced by chrysotiles was similar to those induced by equitoxic doses of H2O2. Moreover, treatment with catalase abrogated the accumulation of y-H2AX, a biomarker of DNA double-strand breaks, induced by chrysotile fibers. CONCLUSIONS: Our results provide novel information on the frequencies and types of mutations induced by asbestos fibers in the gpt delta transgenic mouse mutagenic assay, which shows great promise for evaluating fiber/particle mutagenicity in vivo.

Targeted irradiation of shoot apical meristem of Arabidopsis embryos induces long-distance bystander/abscopal effects.

Bystander effects induced by low-dose ionizing radiation have been shown to occur widely in many cell types and may have a significant impact on radiation risk assessment. Although the region of radiation damage is known to be much greater than the initial target volume irradiated, it remains to be seen whether this response is limited to the specific organ irradiated, spans a limited region of the body, or even covers the whole body of the target. To determine whether long-distance bystander/abscopal effects exist in whole organisms and to clarify the problem of intercellular communication, in the present study a specific cell group, the shoot apical meristem in Arabidopsis embryo, was irradiated with a defined number of protons and examined for root development postirradiation. The results showed that after direct damage to the shoot apical meristem from ion traversals, root hair differentiation, primary root elongation and lateral root initiation were all inhibited significantly in postembryonic development, suggesting that radiation-induced long-distance bystander/abscopal responses might exist in the whole organism. To further scrutinize the mechanism(s) underlying these inhibitory effects, a DR5-GUS transgenic Arabidopsis was used. The results showed that accumulation of the reporter GUS gene transcript in irradiated shoot apical meristem embryos decreased in the postembryonic development. Treatment with either 2,4-dichlorophenoxyacetic acid, a synthetic plant auxin, or DMSO, a effective reactive oxygen species (ROS) scavenger, could rescue the reporter GUS enzyme accumulation and the length of primary root in irradiated shoot apical meristem embryos, indicating that ROS or probably the ROS related auxin and auxin-dependent transcription process may be involved in radiation-induced long-distance bystander/abscopal effects.

Mutagenicity of diesel exhaust particles mediated by cell-particle interaction in mammalian cells.

Diesel exhaust particle (DEP) has been identified as a class 2A human carcinogen and closely related to the increased incidence of respiratory allergy, cardiopulmonary morbidity and mortality, and risk of lung cancer. However, the molecular mechanisms of DEP mutagenicity/carcinogenicity are still largely unknown. In the present study, we focused on the mutagenicity of DEPs in human-hamster hybrid (A(L)) cells and evaluated the role of cell-particle interaction in mediating mutagenic process. We found that DEPs formed micron-sized aggregates in the medium and located mainly in large cytoplasmic vacuoles of cells by 24h treatment. The cellular granularity was increased by DEP treatment in a dose-dependent manner. DEPs resulted in a dose-dependent increase of mutation yield at CD59 locus in A(L) cells, while inflicting minimal cytotoxicity. There was a more than two-fold increase of mutation yield at CD59 locus in A(L) cells exposed to DEPs at a dose of 50mug/ml. Such induction was significantly reduced by concurrent treatment with phagocytosis inhibitors, cytochalasin B and ammonium chloride (p<0.05). These results provided direct evidence that DEPs was mutagenic in mammalian cells and that cell-particle interaction played an essential role in the process.

A dose-survival model for low energy ion irradiation.

PURPOSE: To explain the dose survival data for low energy ion irradiation in bioorganisms. MATERIAL AND METHODS: Applying DNA target theory and considering physical processes of low energy ion radiation, a mathematical dose-survival model was derived to analyze the data of low energy ion irradiation of bioorganisms. RESULTS: The survival fraction S of bioorganisms after a dose D of low energy ion irradiation was: The analysis showed that survival fraction of bioorganisms under low energy ion irradiation can be well fitted with this formula. CONCLUSIONS: It was shown that the dose-survival effects of low energy ion irradiation in bioorganisms were related to the physical processes of low energy ion irradiation and repair effects that can be explained by momentum transfer processes occurring with low energy ion irradiation.

Elevated sodium chloride concentrations enhance the bystander effects induced by low dose alpha-particle irradiation.

Previous studies have shown that high NaCl can be genotoxic, either alone or combined with irradiation. However, little is known about the relationship between environmental NaCl at elevated conditions and radiation-induced bystander effects (RIBE). RIBE, which has been considered as non-targeted bystander responses, has been demonstrated to occur widely in various cell lines. In the present study, RIBE under the elevated NaCl culture condition was assessed in AG 1522 cells by both the induction of gamma-H2AX, a reliable marker of DNA double-strand break (DSB) for the early process (<1h post irradiation), and the generation of micronuclei (MN), a sensitive marker for relative long process of RIBE. Our results showed that in the absence of irradiation, NaCl at elevated concentration such as 8.0, 9.0 and 10.0g/L did not significantly increase the frequency of gamma-H2AX foci-positive cells and the number of foci per positive cell comparing with that NaCl at a normal concentration (6.8g/L). However, with 0.2cGy alpha-particle irradiation, the induced fraction of gamma-H2AX foci-positive cells and the number of induced gamma-H2AX foci per positive cell were significantly increased in both irradiated and adjacent non-irradiated regions. Similarly, the induction of MN by 0.2cGy alpha-particle irradiation also increased with the elevated NaCl concentrations. With N(G)-methyl-l-arginine, an inhibitor of nitric oxide synthase, the induced fraction of foci-positive cells was effectively inhibited both in 0.2cGy alpha-particle irradiated and adjacent non-irradiated regions under either normal or elevated NaCl conditions. These results suggested that the cultures with elevated NaCl medium magnified the damage effects induced by the low dose alpha-particle irradiation and nitric oxide generated by irradiation was also very important in this process.

Nitrogen removal from eutrophic water by floating-bed-grown water spinach (Ipomoea aquatica Forsk.) with ion implantation.

The aim of this study was to investigate the use of water spinach (Ipomoea aquatica Forsk.) with N(+) ion-beam implantation for removal of nutrient species from eutrophic water. The mutated water spinach was grown on floating beds, and growth chambers were used to examine the growth of three cultivars of water spinach with ion implantation for 14 days in simulated eutrophic water at both high and low nitrogen levels. The specific weight growth rates of three cultivars of water spinach with ion implantation were significantly higher than the control, and their NO(3)-N and NH(4)-N removal efficiencies were also greater than those of the control. Furthermore, compared with the control, the nitrogen contents in the plant biomass with ion implantation were higher as well.

[Non-thermal bioeffects of static and extremely low frequency electromagnetic fields].

Since epidemiologic studies have reported a modestly increased risk of oncogenesis associated with certain electromagnetic fields (EMF), popular media and scientists have raised concerns about possible health hazards of environmental exposure to EMF. Laboratory-based experiments have shown that a variety of biological responses were induced by EMF, although these results were controversial and conflicting. The non-thermal effects of low energy EMF,the possible interaction of EMF with biological system have become focus topics in the biolectromagnetic fields. This paper focuses on recent studies of static and extremely low frequency electromagnetic fields, especially the interactive mechanism between EMF and cellular membrane and protein kinase signal transduction pathways. The potential genetic toxicity and risk evaluation are also discussed. However, the existence of some positive findings and the limitations in the set of studies suggest a need for more work.