Inorganic carbon utilization strategies of plateau aquatic plants in response to native habitats.

Aquatic plants are a crucial component of the aquatic ecosystem in the Tibetan Plateau region. Researching the adaptability of plateau aquatic plants in photosynthesis to the plateau environment can enhance understanding of the operational mechanisms of plateau ecosystems, thereby providing a scientific basis for the protection and management of plateau aquatic ecosystems. This study presents an investigation of photosynthetic inorganic carbon utilization strategies and photosynthetic efficiency of 17 aquatic plants under natural growing conditions in Niyang River basin on the Tibetan Plateau. In pH-drift experiments, 10 of 17 species were able to utilize HCO3-, and environmental factors like water pH were shown to have a significant effect on the ability of the tested species to utilize HCO3-. Titratable acidity in the leaves of Stuckenia filiformis, Zannichellia palustris, Batrachium bungei, and Myriophyllum spicatum showed significant diurnal fluctuations at certain sampling sites, indicating the presence of CAM. In B. bungei, water pH positively correlated with CAM activity, while CO2 concentration negatively correlated with CAM activity. The chlorophyll fluorescence analysis revealed that aquatic plants inhabiting the Tibetan Plateau exhibited photosynthetic adaptations. In conclusion, the aquatic plants on the Tibetan Plateau employ diverse strategies for utilizing inorganic carbon during photosynthesis, exhibiting their flexible adaptability to the native high-altitude habitats of the Tibetan Plateau.

Red light mitigates Cd toxicity in Egeria densa by restricting Cd accumulation and modulating antioxidant defense system.

Controlling light qualities have been acknowledged as an effective method to enhance the efficiency of phytoremediation, as light has a significant impact on plant growth. This study examined the effects of light qualities on cadmium (Cd) tolerance in aquatic plant Egeria densa using a combination of biochemical and transcriptomic approaches. The study revealed that E. densa exhibits higher resistance to Cd toxicity under red light (R) compared to blue light (B), as evidenced by a significant decrease in photosynthetic inhibition and damage to organelle ultrastructure. After Cd exposure, there was a significantly reduced Cd accumulation and enhanced levels of both glutathione reductase (GR) activity and glutathione (GSH), along with an increase in jasmonic acid (JA) in R-grown E. densa compared to B. Transcriptional analysis revealed that R caused an up-regulation of Cd transporter genes such as ABCG (G-type ATP-binding cassette transporter), ABCC (C-type ATP-binding cassette transporter), and CAX2 (Cation/H+ exchanger 2), while down-regulated the expression of HIPP26 (Heavy metal-associated isoprenylated plant protein 26), resulting in reduced Cd uptake and enhanced Cd exportation and sequestration into vacuoles. Moreover, the expression of genes involved in phytochromes and JA synthesis was up-regulated in Cd treated E. densa under R. In summary, the results suggest that R could limit Cd accumulation and improve antioxidant defense to mitigate Cd toxicity in E. densa, which might be attributed to the enhanced JA and phytochromes. This study provides a foundation for using light control methods with aquatic macrophytes to remediate heavy metal contamination in aquatic systems.

Red light alleviates Cd toxicity in Egeria densa by modifying carbon-nitrogen metabolism and boosting energy metabolism.

Among the various pollutants detected in aquatic ecosystems, cadmium (Cd) is considered as one of the most hazardous. Freshwater macrophytes have been recognized as possible candidates for eliminating Cd from environment. Nevertheless, the impact of light quality on their ability to tolerate Cd toxicity remains unclear, and the underlying mechanisms have yet to be fully elucidated. In this study, we utilized physiological testing and metabolomics to explore the potential mechanisms by which light quality influences the ability of Egeria densa, a significant Cd hyperaccumulator, to withstand Cd toxicity. The study demonstrated that following Cd treatment, E. densa grown under red light exhibited superior photosynthetic efficiency compared to those grown under blue light, as evidenced by significantly increased photosynthetic rate, higher starch content, and greater activity of photosynthetic enzymes. Moreover, metabolomic analyses revealed that under Cd stress, E. densa grown under red light exhibited an enhanced glycolysis for increased energy production. Sucrose metabolism was also improved to generate sufficient sugar including glucose, fructose and mannose for osmotic adjustment. Moreover, under red light, the heightened production of α-ketoglutarate via tricarboxylic acid (TCA) cycle redirected nitrogen flow towards the synthesis of resilient substances such as γ-Aminobutyric Acid (GABA) and methionine. The production of these substances was ∼2.0 and 1.3 times greater than that of treatment with Cd under blue light, thereby improving E. densa's capacity to withstand Cd stress. This study represents the initial investigation into the possible mechanisms by which light quality influences the ability of E. densa to withstand Cd toxicity through regulating CN metabolism. Furthermore, these findings have the potential to improve phytoremediation strategies aimed at reducing Cd pollution.

Blue Light Enhances Cadmium Tolerance of the Aquatic Macrophyte Potamogeton crispus.

Cadmium (Cd) is highly toxic and widely distributed in aquatic systems due to its high solubility and mobility in water, which can severely inhibit the survival of aquatic macrophytes. The phytotoxicity of Cd depends on environmental factors; however, it remains unclear whether and how light quality affects its toxicity on aquatic macrophytes. In this study, we investigated the effects of Cd on aquatic macrophytes Potamogeton crispus under different light qualities (white, blue, and red light). We evaluated morphological and photo-physiological traits, as well as the cellular antioxidant defense system. Our findings indicate that P. crispus under Cd stress showed notable damage in leaf morphology, decreased photosynthetic efficiency, inhibited HCO3- uptake, and reduced antioxidant enzyme activities, as well as oxidative damage indicated by MDA accumulation and superoxide (O2-) overproduction. However, compared with white or red light under Cd stress, blue light reduced structural damage and oxidative stress caused by Cd while increasing pigment synthesis and photosynthetic efficiency, as well as increasing ascorbate peroxidase (APX) activity. In conclusion, the changes induced by blue light in P. crispus's photosynthesis and antioxidant system strengthen its tolerance to Cd. Further research on signal transmission in relation to light quality in Cd-exposed aquatic plants is still needed.

Long non-coding RNA MIR22HG inhibits the proliferation and migration, and promotes apoptosis by targeting microRNA-9-3p/ SOCS1 axis in small cell lung cancer cells.

BACKGROUND: This study aims to determine the role of long non-coding RNA (LncRNA) MIR22HG in small cell lung cancer (SCLC), and to explore its relevant mechanism. METHODS AND RESULTS: The expressions of genes and proteins in SCLC cells were examined applying qRT-PCR and western blot. Cell proliferation estimation was implemented utilizing cell counting kit-8 (CCK-8) and colony formation assays; the assessment of cell migration and invasion was operated employing Wound healing and Transwell; apoptosis evaluation was conducted adopting flow cytometric assay. Binding relationships was confirmed by luciferase reporter assay. Moreover, SCLC animal model was established to explore the role of MIR22HG in vivo. It was found that MIR22HG was declined and miR-9-3p was elevated in five SCLC cell lines (NCI-H446, NCI-H69, SHP-77, DMS79 and NCI-H345) in comparison with normal human bronchial epithelial cell line (NHBE). More interestingly, overexpression of MIR22HG resulted in decreased cell viability, declined colony formation, diminished capacities of cell migration and invasion in NCI-H446 and NCI-H345 cells but induced more apoptotic cells. However, these impacts were reversed by miR-9-3p upregulation. Meanwhile, MIR22HG could bind to miR-9-3p and negatively regulate its expression in SCLC. What's more, LncRNA MIR22HG overexpression was also testified to elevate SOCS1 via downregulating miR-9-3p expression. Furthermore, in vivo study further confirmed the role of MIR22HG/miR-9-3p in tumor regulation of SCLC. CONCLUSIONS: In conclusion, MIR22HG in SCLC was found to modulate miR-9-3p level and might act as a possible biomarker for SCLC treatment.

Far upstream element -binding protein 1 (FUBP1) participates in the malignant process and glycolysis of colon cancer cells by combining with c-Myc.

Human distal upstream element (Fuse) binding protein 1 (FUBP1) is a transcriptional regulator of c-Myc and represents an important prognostic marker in many cancers. Therefore, the present study aimed to investigate whether FUBP1 could combine with c-Myc to participate in the progression of colon cancer. Detection of FUBP1 expression was done through reverse transcription-quantitative PCR (RT-qPCR), and the combination of FUBP1 and c-Myc was detected by immunoprecipitation assay. Cell counting kit (CCK)-8, colony formation, transwell and wound healing were applied for assessing the ability of cells to proliferate, migrate, and invade; glycolysis and lactic acid detection kits were used to detect glucose uptake and lactic acid content, while western blotting was adopted to detect the protein expression of glycolysis-related genes. FUBP1 expression was elevated in HCT116 cells relative to other colon cancer cell lines, and silencing FUBP1 could inhibit the ability of HCT116 cells to proliferate, migrate, invade and glycolysis, and enhance its apoptosis. In addition, the results of immunoprecipitation experiments showed that FUBP1 could bind to c-Myc. c-Myc overexpression reversed the inhibitory effects of FUBP1 knockdown on the ability of HCT116 cells to proliferate, migrate, invade and glycolysis. The results indicated that FUBP1 could participate in the deterioration process of colon cancer cells by combining with c-Myc, and it has clinical significance for understanding the key role of FUBP1 in tumor genesis.

The inhibitory effects of Dulaglutide on cellular senescence against high glucose in human retinal endothelial cells.

Diabetic nephropathy is one of the most important chronic microvascular complications of diabetes, and its main feature is diabetic glomerulosclerosis. Endothelial sirtuin 1 (SIRT1) expression is related to aging, and reducing SIRT1 expression promotes endothelial cell aging. Plasminogen activator inhibitor-1 (PAI-1) can be synthesized in a variety of cells, such as endothelial cells. Dulaglutide is a glucagon-like peptide-1 (GLP-1) drug, and it can activate the GLP-1 receptor and promote the conversion of intracellular adenosine triphosphate to adenylate cyclase, thereby activating phosphokinase A, and regulating blood glucose levels effectively in the body. We analyzed the effects of Dulaglutide on inhibiting cell senescence by studying the effects of its different concentrations on telomerase activity and senescence-related gene expression. Our results suggest that Dulaglutide can alleviate high-glucose-induced oxidative stress in human retinal endothelial cells by restoring the expressions of SIRT1 and endothelial nitric oxide synthase (eNOS), thereby inhibiting the expression of PAI-1, and restoring telomerase activity. This suggests that the activity of retinal endothelial cells can be controlled by regulating the expression of SIRT1, so as to achieve the effect of treating diabetic retinopathy.

Ischemic postconditioning provides protection against ischemia-reperfusion injury in intestines of rats.

In the present study, we investigated the protective role of ischemic postconditioning (IPOST) against intestine ischemia-reperfusion (I/R) injury in rats. Male Sprague-Dawley rats were divided into sham-operation group (S), I/R group (I/R), ischemic preconditioning group (IPC), ischemic postconditioning group (IPOST). After reperfusion, small intestines were resected for histopathologic evaluations. To evaluate DNA fragmentation, resolving agarose gel electrophoresis was performed. To measure cellular apoptotic rates in intestine tissues, we performed TUNEL staining. To examine lipid peroxidation, production of superoxide radicals and tissue neutrophil infiltration, we tested the content of malondialdehyde and activities of superoxidase dismutase and myeloperoxidase in intestine tissues, respectively. Under light microscope, intestinal mucosal impairment in IPOST and IPC groups was found milder than that in I/R group (P < 0.05). The number of apoptosis cells in I/R group was significantly higher than that in IPOST and IPC groups (P < 0.05). The content of malondialdehyde and activity of myeloperoxidase were significantly reduced in IPOST group and IPC group compared with I/R group, but the activity of superoxidase dismutase in IPOST group and IPC group was enhanced compared with I/R group (P < 0.05). These results suggest that IPOST results in protection against intestine I/R injury, which may be related to reduced production of reactive oxygen species, enhanced activities of antioxidant systems and inhibited apoptosis of intestinal mucosal cells.

Regulation on RhoA in vascular smooth muscle cells under inflammatory stimulation proposes a novel mechanism mediating the multiple-beneficial action of acetylsalicylic acid.

Recent studies have revealed the additional beneficial effects of acetylsalicylic acid (aspirin) in the medication of cardiovascular diseases. The small GTPase RhoA as an important signaling factor is implicated in a wide range of cell functions. This study aimed to investigate the regulatory effect of acetylsalicylic acid on RhoA in vascular smooth muscle cells (VSMCs). We found that aspirin at 300 µM suppressed VSMCs proliferation stimulated by LPS, and this inhibitory effect was partially mediated by inhibiting the iNOS/NO pathway. RhoA overexpression was downregulated by aspirin (both 30 and 300 µM) because of enhanced degradation of RhoA protein. The effect of LPS on increasing active RhoA level was significantly attenuated by aspirin (300 µM), which exerted no effect on RhoA translocation. The promoted RhoA phosphorylation under LPS stimulation, coupled with RhoA protein expression, was greatly decreased by aspirin treatment. No effect of aspirin was found on the expression, activation, and phosphorylation of RhoA in VSMCs devoid of inflammatory stimulation. Our investigation indicates that the regulation of RhoA by aspirin in VSMCs under inflammatory stimulus could be a novel mechanism via which aspirin, apart from the COX-dependent action, exerted the multiple beneficial effects.

Pathological significance of epidermal growth factor receptor expression and amplification in human gliomas.

AIMS: To investigate epidermal growth factor receptor (EGFR) expression and amplification in gliomas and to assess their association with survival. METHODS AND RESULTS: Immunohistochemistry and fluorescence in-situ hybridization were performed to analyse EGFR status in 158 cases of primary glioma. Kaplan-Meier survival and Cox regression analyses were performed to analyse the prognosis of patients. Overexpression of EGFR and expression of EGFR variant III (EGFRvIII) were found in 102 cases (64.6%) and 47 cases (29.7%), respectively. Overexpression of EGFR was significantly correlated with World Health Organization (WHO) grade and Karnofsky performance score (KPS) (both P < 0.05). Expression of EGFRvIII was significantly correlated with WHO grade, gender, age, and KPS (all P < 0.05). EGFR amplification was found in 46 cases (29.1%), and was significantly correlated with WHO grade, age, KPS and EGFR overexpression (all P < 0.05). Cox multifactor analysis showed that EGFR amplification was an independent unfavourable prognostic factor for human gliomas at all ages, and EGFRvIII was an independent prognostic factor in patients older than 60 years. CONCLUSION: EGFR amplification and EGFRvIII expression were associated with an unfavourable prognosis for patients of all ages, and for those older than 60 years, respectively. The differing significance of EGFR status in young and old glioma patients and its impact on prognosis needs further study.

Regulation of HRG-ß1-induced proliferation, migration and invasion of MCF-7 cells by upregulation of GPR30 expression.

The cooperation and communication between different cell signaling transduction pathways are considered critical in the development of various types of cancer as well as drug resistance. There is evidence of crosstalk between the G protein-coupled receptor 30 (GPR30), the newly discovered estrogen receptor (ER), and the ErbB family. Heregulin (HRG)-ß1, the ligand for ErbB3 and ErbB4, upregulates GPR30 expression in MCF-7, T-47D and BT-474 breast cancer cell lines that express ERα. In the present study, recombinant human HRG-ß1 was used to investigate the upregulation of GPR30 expression by HRGs in MCF-7 breast cancer cells which were ERα-positive. In MCF-7 cells, the ErbB2 inhibitor, AG825, the MAPK inhibitor, PD98059, and the MEK1/2 inhibitor, U0126, blocked the HRG-ß1-induced GPR30 expression. 17-ß-estradiol (E2) boosted the HRG-ß1-induced proliferation, migration and invasion of MCF-7 cells. Similar to E2, the specific GPR30 agonist, G-1, promoted HRG-ß1-induced migration and invasion, but inhibited growth. Using the specific GPR30 antagonist, G-15, or the small interfering RNA for GPR30, the functions of GPR30 after treatment with HRG-ß1 were further investegated. The results from our study indicate that the interruption between GPR30 signaling and the ErbB family system may serve as a promising therapeutic strategy for breast cancer.

Heregulin-ß1-induced GPR30 upregulation promotes the migration and invasion potential of SkBr3 breast cancer cells via ErbB2/ErbB3-MAPK/ERK pathway.

Estrogen receptor (ER)-negative breast cancer cells are probably more aggressive with larger metastatic potential than ER-positive cells. Loss of ER in recurrent breast cancer is associated with poor response to endocrine therapy. G protein-coupled receptor 30 (GPR30) is expressed in half of ER-negative breast cancers. Tumor cell-derived heregulin-ß1 (HRG-ß1) is also found mainly in ER-negative cancer. In SkBr3 breast cancer cells that lack ER but express GPR30, HRG-ß1 upregulates mRNA and protein levels of GPR30 by promoting ErbB2-ErbB3 heterodimerization and activating the downstream MAPK-ERK signaling pathway. Moreover, GPR30 boosts HRG-ß1-induced migration and invasion of SkBr3 cells after combinative treatment with E2, 4-hydroxy-tamoxifen or the specific GPR30 agonist G-1, which are blocked by the specific GPR30 antagonist G-15 or the transfection with the small interfering RNA for GPR30. The ErbB2 inhibitor AG825 and the MEK1/2 inhibitor U0126 also partly inhibit the enhanced migration and invasion. Therefore, HRG-ß1-induced migration and invasion partly depend on the upregulation of GPR30 expression through activation of the ErbB2-ERK pathway in SkBr3 cells. The results of this study indicate that the crosstalk between GPR30 and HRGs signaling is important for endocrine therapy resistance and may provide a new therapeutic way to treat breast cancer.

Proteomic analysis of primary colon cancer-associated fibroblasts using the SELDI-ProteinChip platform.

OBJECTIVE: Cancer-associated fibroblasts (CAFs) are one of the hallmarks of the cancer microenvironment. Recent evidence has indicated that CAFs are more competent in enhancing cancer cell growth and migration than normal fibroblasts. However, the unique protein expression of CAFs has not been fully elucidated. This study aims to investigate the characterizations of colon CAFs by comparing the differential protein expression between CAFs and normal fibroblasts. METHODS: Primary fibroblasts were isolated from surgical specimen of human colon cancer and matched normal colonic tissue. Purity of the cell population was verified through immunostain analysis. Total cell lysates and conditioned media from each group of cells were extracted, and protein expression analysis was conducted using the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) ProteinChip platform. RESULTS: Most primary cells showed typical fibroblast-like features after two weeks. Increased proportion of α-smooth muscle actin-positive myofibroblasts was detected within the CAFs in four of the six pairs of primary cells. Fibroblast activation protein was weakly expressed in most cells without differences. Using SELDI-TOF-MS ProteinChip platform, four protein peaks mass over charge ratio (m/z) 1142, 3011, 4035, and 4945 were detected in the total cell lysates, and two protein peaks m/z 1368 and 1389 were detected in the conditioned media. The potential candidate proteins found in the Swiss-Prot database include morphogenetic neuropeptides, FMRFamide-related peptides, insulin-like growth factor II, thymosin ß-4-like protein 3, and tight junction-associated protein 1. CONCLUSIONS: Using the SELDI-ProteinChip platform, differential protein expressions were identified in colon CAFs compared with normal colonic stromal fibroblasts. The complex proteomic alternations in colon CAFs may play important roles related to the colon cancer microenvironment.

Metal-organic frameworks for reverse-phase high-performance liquid chromatography.

Metal-organic framework MIL-53(Al) is explored for reverse-phase high-performance liquid chromatographic separation of a wide range of analytes from non-polar to polar, and acidic to basic solutes with high resolution, good selectivity, stability and reproducibility.

High-performance liquid chromatographic separation of position isomers using metal-organic framework MIL-53(Al) as the stationary phase.

Metal-organic framework MIL-53(Al) was explored as the stationary phase for high-performance liquid chromatographic separation of position isomers using a binary and/or polar mobile phase. Baseline separations of xylene, dichlorobenzene, chlorotoluene and nitrophenol isomers were achieved on the slurry-packed MIL-53(Al) column with high resolution and good precision. The effects of mobile phase composition, injected sample mass and temperature were investigated. The separation of xylene, dichlorobenzene, chlorotoluene and nitrophenol isomers on MIL-53(Al) were controlled by entropy change.

A strong inorganic acid-initiated methacrylate polymerization strategy for room temperature preparation of monolithic columns for capillary electrochromatography.

A facile strong inorganic acid-initiated methacrylate polymerization strategy was developed for fabricating monolithic columns at room temperature. The prepared monoliths were characterized by FTIR spectrometry, mercury intrusion porosimeter and SEM, while their performance was evaluated by CEC for the separation of various types of compounds including alkyl benzenes, polycyclic aromatic hydrocarbons, nonsteroidal anti-inflammatory drugs, anilines, and nitrophenol isomers. The column-to-column and batch-to-batch reproducibility for the prepared monoliths in terms of the RSD of EOF flow velocity, retention factor, and the minimum plate height of naphthalene ranged from 3.4 to 12.4%. The fabricated monoliths gave excellent performance for the separation of the test neutral compounds with the theoretical plates of 170,000-232,000 plates per meter for thiourea, and 77,400-112,300 plates per meter for naphthalene. The proposed strong inorganic acid-initiated methacrylate polymerization strategy is a promising alternative for fabricating organic polymer-based monoliths.

Exploration of coordination polymer as sorbent for flow injection solid-phase extraction on-line coupled with high-performance liquid chromatography for determination of polycyclic aromatic hydrocarbons in environmental materials.

The copper(II) isonicotinate (Cu(4-C5H4N-COO)2(H2O)4) coordination polymer was prepared, characterized and explored as sorbent for flow injection solid-phase extraction on-line coupled with high-performance liquid chromatography (HPLC) for determination of trace polycyclic aromatic hydrocarbons (PAHs) in environmental matrices. Naphthalene, phenanthrene, anthracene, fluoranthene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene and benzo(ghi)perylene with various shape, size and hydrophobicity were used as model analytes. The porosity of the coordination polymer allows these guest PAHs molecules to diffuse into the buck structure, and the shape and size of the pores lead to shape- and size-selectivity over the guests. The precolumn packed with the coordination polymer was shown to be promising for solid-phase extraction of PAHs in environmental samples with subsequent HPLC separation and UV detection. With extraction of 50 ml of sample solution, the enhancement factors for the PAHs studied ranged from 200 to 2337, depending on the shape, size and hydrophobic property of the PAHs. The detection limits (S/N = 3) of 2-14 ng l(-1) and the sample throughput of 3 samples h(-1) were obtained. The developed method was applied to the determination of trace PAHs in a certified reference material (coal fly ash) and local water samples.

Evaluation of expanded graphite as on-line solid-phase extraction sorbent for high performance liquid chromatographic determination of trace levels of DDTs in water samples.

Dichlorodiphenyltrichloroethane (DDT) and its metabolites are a typical kind of persistent organic pollutants (POPs). Development of a simple, cost-effective and sensitive methodology to monitor DDTs concentrations in water environment is of particular significance for understanding the fate and behavior of these pollutants. In this paper, a method on the basis of solid-phase extraction (SPE) using expanded graphite (EG) as sorbent coupled on-line with high performance liquid chromatography (HPLC) was developed for the determination of trace levels of p,p'-DDD (2,2-bis(4-chlorophenyl)-1,1-dichloroethane), p,p'-DDT, o,p'-DDT and p,p'-DDE (2,2-bis(4-chlorophenyl)-1,1-dichloroethene) in water. The analytes in water were preconcentrated onto the SPE column packed with expanded graphite, and subsequently eluted with methanol-water (90:10) mixed solvent. HPLC with a photodiode array detector was used for their separation and detection. The developed on-line solid-phase extraction protocol for HPLC permits the current HPLC separation and the next preconcentration proceeded in parallel, and thus allows one determination within 8min. The precision (R.S.D.) for 10 replicate injections of a mixture of 1mugl(-1) of each analyte was 3.2-6.2% for the peak area measurement. The detection limits (S/N=3) for preconcentrating 50ml of sample solution ranged from 10 to 25ngl(-1) at a sample throughput of 7.5samplesh(-1). The enhancement factors were about 700. The method was applied to the determination of trace p,p'-DDD, p,p'-DDT, o,p'-DDT and p,p'-DDE in local lake, river and tap water samples.

Purification and properties of lipases/esterases from a Bacillus strain for enantioselective resolution of (S)-Ketoprofen.

A Bacillus strain having a certain extent of asymmetric resolution for (S)-ketoprofen was studied for its culture and conversion condition. The distribution and properties of lipases/esterases from its bacterial cell and the change of asymmetric resolution ability were also investigated. After purification by hydrophobic interaction chromatography and gel filtration, the lipase/esterase terase activity of active fraction was 61.7 times higher than the crude enzyme and the purity of 23 kDa protein increased more than 400 times. Hydrolytic activity of lipase/esterase using ketoprofen chloroethyl ester and pNPA as substrate remained essentially constant in the reaction system during the purification procedure. The above result clearly indicated that the application of hydrophobic interaction chromatography-gel filtration was fast, useful and effective when compared to the difficulties in purification of low content crude enzyme prepared directly from bacterial cells. Experimental results also indicated that there are several lipases/esterases in the bacillus cell selective to the optically active ketoprofen and the conversion result was derived from the comprehensive function of all these enzymes. This is a new way to understand the bacterial asymmetric resolution with high conversion rate and low optical purity. The ESI-QUAD-TOF mass spectrum was used to analyze 23 kDa protein. Protein identities were revealed by searching sequence databases (MSDB) with peptide sequence tags. The result showed that 23 kDa protein is a novel protein.

On-line coupling of flow injection displacement sorption preconcentration to high-performance liquid chromatography for speciation analysis of mercury in seafood.

A simple and cost-effective method for speciation analysis of trace mercury in seafood was developed by on-line coupling flow injection microcolumn displacement sorption preconcentration to high-performance liquid chromatography (HPLC) with UV detection. The methodology involved the presorption of the Cu-PDC (pyrrolidine dithiocarbamate) chelate onto a microcolumn packed with a cigarette filter sorbent, simultaneous preconcentration of Hg(II), methylmercury (MeHg), ethylmercury (EtHg), and phenylmercury (PhHg) onto the microcolumn through a displacement reaction with the presorbed Cu-PDC, and their subsequent elution from the microcolumn for on-line HPLC separation. Interferences from heavy metal ions with lower stability of their PDC chelates relative to Cu-PDC were minimized without the need of any masking agents. With the consumption of 4.0 ml of sample solution, the enrichment factors were about 80. The detection limits were 10-25 ng g(-1) (as Hg) in fresh tissue. Precision (R.S.D. (%), n = 5) ranged from 2 to 3% at the 500 microg l(-1) (as Hg) level. The developed technique was validated by analyzing a certified reference material (DORM-2, dogfish-muscle), and was shown to be useful for mercury speciation in real seafood samples.