Diatom cell-size composition as a novel tool for quantitative estimates of the water table in peatlands.

Diatom cell-size composition is an indicator of aquatic environmental changes but has been rarely investigated, especially in semi-terrestrial peatlands. In this study, both taxonomic composition and cell-size composition of diatoms were analysed in 41 samples from two montane peatlands, northeastern China. Redundancy analyses revealed that diatom taxonomic composition was significantly related to the depth to the water table (DWT) and Ca2+, while cell-size composition was significantly associated with DWT and Si. DWT was the most important factor and its sole effect explained 26.2% and 17.9% of the total variance in taxonomic composition and cell-size composition, respectively. Accordingly, diatom-based water-table transfer functions were developed based on taxonomic composition and cell-size composition, respectively. The maximum-likelihood (ML) model based on diatom taxonomic composition had the best performance, with a correlation coefficient value (R2) of 0.78 and the root mean squared error of prediction (RMSEP) of 6.66 cm. The ML model based on cell-size composition had similar performance, with an R2 of 0.78 and the RMSEP of 6.87 cm, suggesting that diatom cell-size composition can be a new quantitative means to track past water-table changes. This method requires further appraisal with palaeoecological data but offers a new option that deserves exploration.

Clinicopathological features, prognostic factors, and prognostic survival prediction in patients with extrahepatic bile duct cancer liver metastasis.

PURPOSE: Extrahepatic bile duct cancer (EBDC) is a compound malignant tumor mainly consisting of extrahepatic cholangiocarcinoma and gallbladder carcinoma. Most EBDC patients are diagnosed at an advanced stage characterized by distant metastases, and the liver is one of the common sites of metastasis. Hence, the purpose of this study is to investigate the clinicopathological features, identify prognostic risk factors, and assess the long-term prognosis of extrahepatic bile duct cancer liver metastasis (EBDCLM). METHODS: We identified 1922 eligible EBDCLM patients from the SEER database.Cox regression models were used to predict independent prognostic factors for overall survival (OS) and cancer-specific survival (CSS),and Kaplan-Meier survival curves were drawn. A nomogram was constructed based on the results of multivariate Cox analysis, and the predictive effect of the nomogram was evaluated. RESULTS: Age, surgery, chemotherapy, brain metastasis, and lung metastasis were common independent prognostic factors for OS and CSS, and radiotherapy and bone metastasis were independent prognostic factors for CSS. The Kaplan-Meier survival curves showed a significant increase in survival for patients aged less than or equal to 70 years, undergoing surgery and chemotherapy, and without lung metastases. The results showed that the nomogram constructed by us had good predictability and ha d strong clinical application value. CONCLUSION: Our study identified age, surgery, chemotherapy, brain metastasis, and lung metastasis as independent prognostic factors for EBDCLM patients. The nomogram can accurately predict the survival probability, which is helpful for clinicians to assess the prognosis of patients with advanced EBDC and provide personalized clinical decisions.

Cryopreservation of Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells at the Optimal Stage.

Retinal pigment epithelial (RPE) cells derived from human embryonic stem cells (hESCs) are superior cell sources for cell replacement therapy in individuals with retinal degenerative diseases; however, studies on the stable and secure banking of these therapeutic cells are scarce. Highly variable cell viability and functional recovery of RPE cells after cryopreservation are the most commonly encountered issues. In the present protocol, we aimed to achieve the best cell recovery rate after thawing by selecting the optimal cell phase for freezing based on the original experimental conditions. Cells were frozen in the exponential phase determined by using the 5-ethynyl-2'-deoxyuridine labeling assay, which improved cell viability and recovery rate after thawing. Stable and functional cells were obtained shortly after thawing, independent of a long differentiation process. The methods described here allow the simple, efficient, and inexpensive preservation and thawing of hESC-derived RPE cells. Although this protocol focuses on RPE cells, this freezing strategy may be applied to many other types of differentiated cells.

Water table level controls methanogenic and methanotrophic communities and methane emissions in a Sphagnum-dominated peatland.

Peatlands are important sources of the greenhouse gas methane emissions equipoised by methanogens and methanotrophs. However, knowledge about how microbial functional groups associated with methane production and oxidation respond to water table fluctuations has been limited to date. Here, methane-related microbial communities and the potentials of methane production and oxidation were determined along sectioned peat layers in a subalpine peatland across four Sphagnum-dominated sites with different water table levels. Methane fluxes were also monitored at these sites. The results showed that mcrA gene copies for methanogens were the highest in the 10- to 15-cm peat layer, which was also characterized by the maximum potential methane production (24.53 ± 1.83 nmol/g/h). Copy numbers of the pmoA gene for type Ia and Ib methanotrophs were enriched in the 0-5 cm peat layer with the highest potential methane oxidation (43.09 ± 3.44 nmol/g/h). For the type II methanotrophs, the pmoA gene copies were higher in the 10- to 15-cm peat layer. Hydrogenotrophic methanogens and type II methanotrophs dominated the methane functional groups. Deterministic process contributed more to methanogenic and methanotrophic community assemblages in comparison with stochastic process. The level of water table significantly shaped methanogenic and methanotrophic community structures and regulated methane fluxes. Compared with vascular plants, Sphagnum mosses significantly reduced the methane emissions in peatlands. Collectively, these findings enhance a comprehensive understanding of the effect of the water table level on methane functional groups, with consequential implications for reducing methane emissions within peatland ecosystems.IMPORTANCEThe water table level is recognized as a critical factor in regulating methane emissions, which are largely dependent on the balance of methanogens and methanotrophs. Previous studies on peat methane emissions have been mostly focused on spatial-temporal variations and the relationship with meteorological conditions. However, the role of the water table level in methane emissions remains unknown. In this work, four representative microhabitats along a water table gradient in a Sphagnum-dominated peatland were sampled to gain an insight into methane functional communities and methane emissions as affected by the water table level. The changes in methane-related microbial community structure and assembly were used to characterize the response to the water table level. This study improves the understanding of the changes in methane-related microbial communities and methane emissions with water table levels in peatlands.

Transcriptome Analysis of Protection by Dendrobium Nobile Alkaloids (DNLA) against Chronic Alcoholic Liver Injury in Mice.

Objective: To investigate the protective effects of Dendrobium nobile Lindl. alkaloids (DNLA) against chronic alcoholic liver injury. C57BL/6J mice were fed with the Lieber−DeCarli alcohol diet to induce chronic alcoholic liver injury. DNLA (20 mg/kg/day) was gavaged along with the alcohol diet for 28 days. Liver injury was evaluated by serum enzymes. Triglyceride levels, histopathology, and transcriptome changes were examined by RNA-Seq and qPCR. DNLA decreased serum triglyceride levels in mice receiving alcohol. Hepatocyte degeneration and steatosis were ameliorated by DNLA, as evidenced by H&E and Oil-red O staining. DNLA brought the alcohol-induced aberrant gene expression pattern towards normal. Alcohol induced 787 differentially expressed genes (padj < 0.01). DNLA induced 280 differentially expressed genes to a much less extent. Ingenuity pathway analysis showed that DNLA ameliorated alcohol-induced oxidative stress and xenobiotic metabolism disruption. qPCR verified that DNLA alleviated over-activation of Cyp2a4, Cyp2b10, and Abcc4; attenuated oxidative stress (Hmox1, Gstm3, Nupr1), reduced the expression of Nrf2 genes (Nqo1, Gclc, Vldlr); and rescued some metabolic genes (Insig1, Xbp1, Socs3, Slc10a2). In conclusion, DNLA was effective against alcohol-induced fatty liver disease, and the protection may be attributed to alleviated oxidative stress and restored metabolism homeostasis, probably through modulating nuclear receptor CAR-, PXR-, and Nrf2-mediated gene expression pathways.

Determining the optimal stage for cryopreservation of human embryonic stem cell-derived retinal pigment epithelial cells.

BACKGROUND: Human embryonic stem cell-derived retinal pigment epithelial cells (hESC-derived RPE) are a promising source for cell-replacement therapy to treat retinal degenerative diseases, but research on RPE cryopreservation is limited. This study aimed to determine the best phase for RPE cryopreservation to preserve the post-thaw function and uncover the mechanism underlying RPE freezing tolerance. METHODS: hESC-derived RPE cells were cryopreserved at various time points after seeding. After thawing, the survival and attachment rates, RPE marker gene expression, apical-basal polarity, PEDF secretion, transepithelial resistance, and phagocytotic ability of post-thaw RPE cells were evaluated. RNA sequencing was performed on RPE cells at three-time points, differentially expressed genes were identified, and gene ontology, Kyoto encyclopedia of genes and genomes, and protein-protein interaction analyses were used to investigate the key pathways or molecules associated with RPE cell freezing tolerance. RESULTS: RPE frozen at passage 2 day 5 (P2D5) had the highest cell viability and attachment after thawing. They also retained properly localized expression of RPE marker genes and biological functions such as PEDF secretion, high transepithelial resistance, and phagocytic ability. The RNA-sequencing analysis revealed that RPE cells at P2D5 expressed high levels of cell cycle/DNA replication and ECM binding associated genes, as well as THBS1, which may serve as a possible hub gene involved in freezing tolerance. We also confirmed that the RPE cells at P2D5 were in the exponential stage with active DNA replication. CONCLUSIONS: We propose that freezing hESC-derived RPE cells during their exponential phase results in the best post-thawing outcome in terms of cell viability and preservation of RPE cell properties and functions. The high expression levels of the cell cycle and ECM binding associated genes, particularly THBS1, may contribute to better cell recovery at this stage.

RNA-Seq Analysis of Protection against Chronic Alcohol Liver Injury by Rosa roxburghii Fruit Juice (Cili) in Mice.

Rosa roxburghii Tratt. fruit juice (Cili) is used as a medicinal and edible resource in China due to its antioxidant and hypolipidemic potentials. The efficacy of Cili in protecting alcohol-induced liver injury and its underlying mechanism was investigated. C57BL/6J mice received a Lieber-DeCarli liquid diet containing alcohol to produce liver injury. After the mice were adapted gradually to 5% alcohol, Cili (4 mL and 8 mL/kg/day for 4 weeks) were gavaged for treatment. The serum enzyme activities, triglyceride levels, histopathology and Oil-red O staining were examined. The RNA-Seq and qPCR analyses were performed to determine the protection mechanisms. Cili decreased serum and liver triglyceride levels in mice receiving alcohol. Hepatocyte degeneration and steatosis were improved by Cili. The RNA-Seq analyses showed Cili brought the alcohol-induced aberrant gene pattern towards normal. The qPCR analysis verified that over-activation of CAR and PXR (Cyp2a4, Cyp2b10 and Abcc4) was attenuated by Cili. Cili alleviated overexpression of oxidative stress responsive genes (Hmox1, Gsta1, Gstm3, Nqo1, Gclc, Vldlr, and Cdkn1a), and rescued alcohol-downregulated metabolism genes (Angptl8, Slc10a2, Ces3b, Serpina12, C6, and Selenbp2). Overall, Cili was effective against chronic alcohol liver injury, and the mechanisms were associated with decreased oxidative stress, improved lipid metabolism through modulating nuclear receptor CAR-, PXR-and Nrf2-mediated pathways.

Improvement of human embryonic stem cell-derived retinal pigment epithelium cell adhesion, maturation, and function through coating with truncated recombinant human vitronectin.

AIM: To explore an xeno-free and defined coating substrate suitable for the culture of H9 human embryonic stem cell-derived retinal pigment epithelial (hES-RPE) cells in vitro, and compare the behaviors and functions of hES-RPE cells on two culture substrates, laminin521 (LN-521) and truncated recombinant human vitronectin (VTN-N). METHODS: hES-RPE cells were used in the experiment. The abilities of LN-521 and VTN-N at different concentrations to adhere to hES-RPE cells were compared with a high-content imaging system. Quantitative real-time polymerase chain reaction was used to evaluate RPE-specific gene expression levels midway (day 10) and at the end (day 20) of the time course. Cell polarity was observed by immunofluorescent staining for apical and basal markers of the RPE. The phagocytic ability of hES-RPE cells was identified by flow cytometry and immunofluorescence. RESULTS: The cell adhesion assay showed that the ability of LN-521 to adhere to hES-RPE cells was dose-dependent. With increasing coating concentration, an increasing number of cells attached to the surface of LN-521-coated wells. In contrast, VTN-N presented a strong adhesive ability even at a low concentration. The optimal concentration of LN-521 and VTN-N required to coat and adhesion to hES-RPE cells were 2 and 0.25 µg/cm2, respectively. Furthermore, both LN-521 and VTN-N could facilitate adoption of the desired cobblestone cellular morphology with tight junction and showed polarity by the hES-RPE cells. However, hES-RPE cells cultivated in VTN-N had a greater phagocytic ability, and it took less time for these hES-RPE cells to mature. CONCLUSION: VTN-N is a more suitable coating substrate for cultivating hES-RPE cells.

The elemental enrichments at Dajiuhu Peatland in the Middle Yangtze Valley in response to changes in East Asian monsoon and human activity since 20,000 cal yr BP.

Here we present multiproxy inorganic geochemical records from a peat core (ZK5) from the Dajiuhu Basin in central China to investigate peatland deposition processes and atmospheric metal pollution and to explore their relationships with East Asian monsoon change and human activities in the Middle Yangtze Valley since 20,000 cal yr BP. The peat physicochemical data including total organic carbon (TOC), trace elements, and grain-size show that the site has changed from a lake during the cold-wet Last Glacial Maximum (LGM; 20,000-18,000 cal yr BP), to a marshy wetland during the mild last deglaciation (18,000-11,500 cal yr BP) and a peatland during the mostly warm and dry Holocene (11,500 cal yr BP-present). This general sequence corresponds with changes in East Asian monsoon indicated by stalagmites δ18O records and boreal summer insolation. Marked decreases in trace element concentrations correspond to two periods of peatland expansion during the abrupt hydroclimatic transitions from the LGM to the last deglaciation and from the last deglaciation to the early Holocene. Warm-dry mid-Holocene might induce high organic matter decomposition in peat sediments. Increasing natural element concentrations since the late Holocene are correlated with the weakening of the summer monsoon and elevated atmospheric dust deposition. Increasing Cu and Pb concentrations in peat record indicate large-scale Cu smelting during the Bronze Age and excessive coal burning during the 10th century or so. The anthropogenic heavy metals were transported by prevailing East Asian summer monsoon and deposited in the Dajiuhu Basin during periods of heightened human activities. Our compilation of heavy metals records across China confirmed the noticeable impacts of the historical human activity on deposition environments during the late Holocene. Consequently, trace elements from the Dajiuhu Basin are reliable proxies for capturing monsoon climate-induced peatland deposition response and present important evidence for a historical atmospheric heavy metal pollution in the Middle Yangtze Valley. Our results offer useful references for peatland evolution and protection under the background of global change.

Geochemical cycle of mercury associated with wet deposition and inflows in a subalpine wetland.

Subalpine wetland is a mercury (Hg) sensitive ecosystem, but there is poor understanding of Hg behavior in this typical wetland. Here, distribution and speciation of Hg in waters of a subalpine wetland (Dajiuhu) in China were investigated, and an initial model of the Hg geochemical cycle in the wetland was established based on Hg mass balance calculations. Concentrations of both total Hg (THg, 9.52 ± 6.61 ng L-1) and total methyl mercury (TMeHg, 0.34 ± 0.44 ng L-1) in the waters during the wet season were higher than in the dry season. The majority of THg was in dissolved form whereas most TMeHg was in particle form. The geochemical models suggested that, due to the wet deposition and surface runoff, the input of THg and TMeHg into the wetland in the wet season (222 and 2.74 g year-1, respectively) was higher than that in the dry season (57.9 and 1.15 g year-1, respectively). The output of THg and TMeHg from the wetland underground runoff in the wet season was estimated to be 154 and 2.51 g year-1, respectively, and in the dry season was 15.9 and 0.43 g year-1, respectively. Other losses of Hg were due to volatilization of Hg0 from the sediment water (30.5 and 12.5 g year-1 in the wet and dry seasons, respectively). The flux of the settling of particulate Hg in the wet season was higher than that in the dry season. The fluxes of Hg diffusion from the porewater were relatively low in comparison to the fluxes of inflows and wet deposition. The flux of oxidation was higher than reduction, while the flux of methylation was higher than demethylation. These results indicated that the elevated levels of THg and MeHg in the Dajiuhu wetland are a consequence of rainfall and surface runoff inputs.

MicroRNA-758 acts as a tumor inhibitor in colorectal cancer through targeting PAX6 and regulating PI3K/AKT pathway.

Recently, a number of microRNAs (miRNAs) have been reported to play different roles in human cancers, including colorectal cancer (CRC). However, the specific role of miR-758 has not been clarified in CRC. Therefore, the aim of the present study was to explore the role of miR-758 in CRC. RT-qPCR and western blot analysis were used to quantify the expression of miR-758 and genes. The function of miR-758 in CRC was investigated using Transwell, CCK-8 and luciferase reporter assays. According to the results, the downregulation of miR-758 expression was associated with aggressive behavior and poor prognosis in CRC patients. miR-758 was shown to restrain the cell viability and metastasis in CRC. In addition, it was confirmed that miR-758 directly targets PAX6 and inhibits CRC progression through targeting PAX6. The results also revealed that miR-758 blocked EMT and PI3K/AKT pathway in CRC. In conclusion, miR-758 acts as a tumor suppressor in CRC by downregulating PAX6.

Characteristics and compound-specific carbon isotope compositions of sedimentary lipids in high arsenic aquifers in the Hetao basin, Inner Mongolia.

Organic matter, as an electron donor, plays a vital role in As mobilization mediated by microorganisms during reductive dissolution of Fe/Mn oxides in shallow aquifers. However, the specific types and sources of organic matter involved in biogeochemical processes accelerating As mobilization are still controversial. Both sediment and groundwater samples were collected at different depths from aquifers of the Hetao Basin, a typical inland basin hosting high As groundwater. Sedimentary lipids and their compound-specific carbon isotope ratios were analyzed to evaluate characteristics and sources of organic matter. Results show that sedimentary As were well correlated with Fe and Mn oxides, suggesting that As exist as Fe/Mn oxide bound forms. Groundwater As far exceeded the drinking water guide value of 10 µg/L. Moreover, As concentrations in shallow groundwater were relatively higher. Lipids in clay were mainly originated from terrestrial higher plants, while that in fine sand samples were derived from terrestrial higher plants, microorganism and petroleum. Shallow fine sand samples were also characterized by evident in-situ biodegradation. Compound-specific carbon isotope compositions of sedimentary lipids showed that short-chain n-alkanes and n-alkanoic acids had more positive δ13C values compared to long-chain compounds, especially in shallow fine sand samples. δ13CTOC were also low in shallow fine sand samples. These results jointly indicate that these lipids in shallow fine sand samples acted as carbon source for indigenous microorganism and the short-chain components were particularly more vulnerable to biodegradation, which may contribute to high As concentrations in shallow groundwater. The new findings provide the first evidence that short chain length n-alkyl compounds afforded a source of potential electron donors for microbially mediated As mobilization process in the shallow aquifers.

Response of carbon cycle to drier conditions in the mid-Holocene in central China.

The nature and extent to which hydrological changes induced by the Asian summer monsoon affected key biogeochemical processes remain poorly defined. This study explores the relationship between peatland drying and carbon cycling on centennial timescales in central China using lipid biomarkers. The difference between peat n-alkane δ2H and a nearby stalagmite δ18O record reveals that intervals of prominent peatland drying occurred during the mid-Holocene. Synchronous with these drier conditions, leaf wax δ13C values show large negative excursions, with the utilization of CO2 respired from the peatland subsurface for plant photosynthesis being a possible mechanism. Crucially, successive drying events appear to have had a cumulative impact on the susceptibility of peat carbon stores to climate change. Concurrently, bacterially derived hopane δ13C values suggest the occurrence of enhanced methane oxidation during the drier periods. Collectively, these observations expand our understanding of how respiration and degradation of peat are enhanced during drying events.

Crystal structure and luminescent properties of novel coordination polymers constructed with bifurandicarboxylic acid.

Four novel coordination polymers (CPs) assembled with [2,2'-bifuran]-5,5'-dicarboxylic acid (H2L) and metal ions ZnII, CdII or CoII have been successfully synthesized. Their molecular structures have been determined to be Zn(L)(H2O)2 (1), [Zn(L)(DMF)]2 (2), [Cd(L)(EtOH)(DMF)2]2 (3) and Co(L)(H2O)4·(H2O)2 (4) by crystallographic analysis. It is found that (2) exhibits a two-dimensional infinite framework, whereas the others exhibit a one-dimensional framework. Both (2) and (3) exhibit ligand-based emission which is blue-shifted compared with the emission from the H2L ligand. Time-resolved spectroscopic measurement reveals that the complexes exhibit a similar lifetime with that of the ligand, which confirms that the emission of the complexes originates from the ligand.

Correction: Molecular Paleoclimate Reconstructions over the Last 9 ka from a Peat Sequence in South China.

[This corrects the article DOI: 10.1371/journal.pone.0160934.].

Molecular Paleoclimate Reconstructions over the Last 9 ka from a Peat Sequence in South China.

To achieve a better understanding of Holocene climate change in the monsoon regions of China, we investigated the molecular distributions and carbon and hydrogen isotope compositions (δ13C and δD values) of long-chain n-alkanes in a peat core from the Shiwangutian (SWGT) peatland, south China over the last 9 ka. By comparisons with other climate records, we found that the δ13C values of the long-chain n-alkanes can be a proxy for humidity, while the δD values of the long-chain n-alkanes primarily recorded the moisture source δD signal during 9-1.8 ka BP and responded to the dry climate during 1.8-0.3 ka BP. Together with the average chain length (ACL) and the carbon preference index (CPI) data, the climate evolution over last 9 ka in the SWGT peatland can be divided into three stages. During the first stage (9-5 ka BP), the δ13C values were depleted and CPI and Paq values were low, while ACL values were high. They reveal a period of warm and wet climate, which is regarded as the Holocene optimum. The second stage (5-1.8 ka BP) witnessed a shift to relatively cool and dry climate, as indicated by the more positive δ13C values and lower ACL values. During the third stage (1.8-0.3 ka BP), the δ13C, δD, CPI and Paq values showed marked increase and ACL values varied greatly, implying an abrupt change to cold and dry conditions. This climate pattern corresponds to the broad decline in Asian monsoon intensity through the latter part of the Holocene. Our results do not support a later Holocene optimum in south China as suggested by previous studies.

Compound-specific carbon isotopic fractionation during transport of phthalate esters in sandy aquifer.

The present paper aims to evaluate the carbon isotopic fractionation of phthalate esters (PAEs) during transport in an sandy aquifer. Breakthrough curves of di-methyl phthalate (DMP), di-ethyl phthalate (DEP), and di-n-butyl phthalate (DBP) in mixed solution were determined by miscible displacement experiment, and simulated using HYDRUS-1D software. The stable carbon isotopes (δ(13)C) of 3 PAEs in effluent were analyzed at different times. Results showed that, in the transport process in sandy sediment, PAEs are mainly trapped into the pore space instead of being adsorbed on the surface of particles. At the initial stage of transport, PAEs with lighter carbon tend to run faster in the sandy sediment, and PAEs with heavier carbon run after. However, there is no priority for the transport of PAEs with different carbon isotopes at Stage Ⅱ with mainly time-limited sorption. So the transport-based isotope fractionation occurs in the front area of contaminant plume. This effect may be relevant for interpreting carbon isotope signatures in the real contaminant site.

Structural insights on mouse L-threonine dehydrogenase: A regulatory role of Arg180 in catalysis.

Mouse L-threonine dehydrogenase (mTDH), which belongs to the short-chain dehydrogenase/reductase (SDR) superfamily and mediates threonine catabolism, plays pivotal roles in both powerful biosynthesis and signaling in mouse stem cells and has a regulatory residue Arg180. Here we determined three crystal structures of mTDH: wild-type (WT) in the apo form; in complex with NAD(+) and a substrate analog, glycerol, or with only NAD(+); as well as the R180K variant with NAD(+). This is the first description of a structure for mammalian SDR-type TDH. Structural comparison revealed the structural basis for SDR-type TDH catalysis remains strictly conserved in bacteria and mammals. Kinetic enzyme assays, and isothermal titration calorimetry (ITC) measurements indicated the R180K mutation has little effect on NAD(+) binding affinity, whereas affects the substrate's affinity for the enzyme. The crystal structure of R180K with NAD(+), biochemical and spectroscopic studies suggested that the R180K mutant should bind NAD(+) in a similar way and have a similar folding to the WT. However, the R180K variant may have difficulty adopting the closed form due to reduced interaction of residue 180 with a loop which connects a key position for mTDH switching between the closed and open forms in mTDH catalysis, and thereby exhibited a significantly decreased kcat/Km value toward the substrate, L-Thr. In sum, our results suggest that activity of GalE-like TDH can be regulated by remote interaction, such as hydrogen bonding and hydrophobic interaction around the Arg180 of mTDH.

Carbon isotopic fractionation during biodegradation of phthalate esters in anoxic condition.

Here we evaluate the quantitative relationship between carbon isotopic fractionation and anoxic biodegradation of phthalate esters (PAEs), a kind of endocrine disruptors. The stable carbon isotope delta values (δ(13)C) of 4 PAEs, i.e. di-methyl phthalate (DMP), di-ethyl phthalate (DEP), di-n-butyl phthalate (DBP), and di-iso-butyl phthalate (DiBP), were analyzed during biodegradation by a pure bacteria strain isolated from the shallow aquifer sediment in anoxic condition. Results showed that the carbon isotopic fractionation in the initial degradation of PAEs was well-described by the Rayleigh equation model with R(2) from 0.8885 to 0.9821. The carbon isotopic fractionation (ε) for DMP and DEP were -4.6±0.4‰ and -2.9±0.1‰, respectively, while DBP and DiBP showed limited isotopic fractionation. A linear relationship between ε values and the total carbon atoms present in straight-carbon-chain PAE molecules with R(2) of 0.9918. The apparent kinetic isotope effects (AKIEs) were calculated for proposed 4 initial transformation pathways of PAEs. The high carbon AKIEs of 1.048 and 1.036 were obtained for single enzymatic hydrolysis of DMP and DEP, respectively, and fell in the expected KIE range of 1.03-1.09. However, the intrinsic carbon isotope effects for enzymatic hydrolysis of DBP and DiBP might be masked.

Electrochemical behavior of neomycin at DNA-modified gold electrodes.

Deoxyribonucleic acid (DNA) modified gold electrodes are prepared by the dry adsorptive method and the electrochemical behavior of neomycin and the influence of Pb(II) are studied by cyclic voltammetry, chronocoulometry, differential pulse voltammetry. It is found that in 0.01M phosphate-buffered saline (PBS) buffer solutions (pH 7.3) at DNA/Au electrode neomycin exhibits an irreversible cathodic peak (E(p)=0.489V), which is more positive and less sensitive compared with that at bare gold electrodes (E(p)=0.423V). In the presence of Pb(II) the peak shifts toward positive with its height increasing. Moreover, the peak height is linear to neomycin concentration over the range of 0.15-57microM. The interaction of Pb(II)-neomycin complex with calf thymus DNA is also studied by calculating the binding constants (K) of the Pb(II)-neomycin complex to DNA and binding site size (s) from voltammetric data (1.0x10(7)M(-1) and 4bp, respectively).