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Biomedical subjects

P He

Publications and source records attributed to P He.

At least 19 recordsLinked to original sources

Roles of mitogen-activated protein kinase pathways during Escherichia coli-induced apoptosis in U937 cells.

Escherichia coli (E. coli) infections play an important and growing role in the clinic. In the present study, we investigated the involvement of members of the mitogen-activated protein kinase (MAPK) superfamily, including extracellular signal-regulated kinase (ERK), c-Jun NH(2)-terminal kinase (JNK) and p38 MAPK, and caspase-3 and 9 activity in E. coli-induced apoptosis in human U937 cells. We found that E. coli induces apoptosis in U937 cell lines in a dose- and time-dependent manner, p38 MAPK and JNK were activated after 10 min of infection with E. coli. In contrast, ERK1/2 was down-regulated in a time-dependent manner. The levels of total (phosphorylation state-independent) p38 MAPK, JNK and ERK1/2 did not change in E. coli-infected U937 cells at all times examined. Moreover, exposure of U937 cells to E. coli led to caspase-3 and 9 activity. For the evaluation of the role of MAPKs, PD98059, SB203580 and SP600125 were used as MAPKs inhibitors for ERK1/2, p38 MAPK and JNK. Inhibition of ERK1/2 with PD98059 caused further enhancement in apoptosis and caspase-3 and 9 activity, while a selective p38 MAPK inhibitor, SB203580 and JNK inhibitor, SP600125 significantly inhibited E. coli-induced apoptosis and caspase-3 and 9 activity in U937 cells. The results were further confirmed by the observation that the caspase inhibitors Z-DEVD-FMK and Z-LEHD-FMK blocked E. coli-induced U937 apoptosis. Taken together, we have shown that E. coli increase p38 MAPK and JNK and decrease ERK1/2 phosphorylation and increase caspase-3 and 9 activity in U937 cells.

Apoptosis↗

Cloning and molecular characterization of a gene coding D-xylulokinase (CmXYL3) from Candida maltosa.

AIMS: To clone and identify a gene (CmXYL3) coding D-xylulokinase from Candida maltosa Xu316 and understand its physiological function. METHODS AND RESULTS: Based on the conserved regions of the known D-xylulokinase-encoding genes, a pair of degenerate primers was designed to clone the CmXYL3 gene from C. maltosa Xu316. The coding region and sequences flanking the CmXYL3 gene were obtained by PCR-based DNA walking method. Southern blotting analysis suggested that there is a single copy of the CmXYL3 gene in the genome. The open reading frame starting from ATG and ending with TAG stop codon encoded 616 amino acids with a calculated molecular mass of 68889.743 Da. The CmXYL3 gene under the control of the GPD1 promoter was heterologously expressed in Saccharomyces cerevisiae deficient in D-xylulokinase (deltaScXKS1::LEU2) activity, and restored growth on D-xylulose. The specific activity of D-xylulokinase varied during xylose fermentation and was correlated with aeration level. After growth on different pentoses and pentitols as sole carbon sources, the highest specific activity of D-xylulokinase was observed on D-xylose. CONCLUSIONS: The CmXYL3 gene isolated from C. maltosa Xu316 encodes a novel D-xylulokinase that plays a pivotal role in xylulose metabolism. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that describes the isolation and cloning of D-xylulokinase gene (CmXYL3) from C. maltosa Xu316. D-xylulokinase is pivotal for growth and product formation during xylose metabolism. Better understanding of the biochemical properties and the physiological function of D-xylulokinase will contribute to optimizing fermentation conditions and determining the strategies for metabolic engineering of C. maltosa Xu316 for further improvement of xylitol yield and productivity.

Amino Acid Sequence↗

Screening and characterization of yeasts for xylitol production.

AIMS: To discover novel naturally occurring xylitol producing yeast species with potential for industrial applications. METHODS AND RESULTS: Exactly 274 strains were cultivated on both solid and liquid screening medium with xylose as the sole carbon resource. Five strains were selected on the basis of significant growth and high degree of xylose assimilation. Their phylogenetic position was confirmed by the PCR-RFLP and sequence analysis of the D1/D2 domain of the 5' end of the large subunit rDNA gene (5'-LSU rDNA). Enzymatic analysis was conducted to compare xylose metabolism in each strain. Candida guilliermondii Xu280 and Candida maltosa Xu316 were found to have high xylose consumption rates and xylitol yields in the batch fermentation under micro-aerobic condition. The effect of the different media with high initial xylose concentration on biosynthesis of xylitol by both strains was investigated. CONCLUSIONS: We have identified Candida spp. strains, which exhibit high levels of xylitol production from xylose suggesting that these may have potential for industrial applications. SIGNIFICANCE AND IMPACTS OF THE STUDY: Microbial species are of importance for xylitol production. Xylitol production involves complicated metabolic regulation including xylose transport, production of key enzymes and cofactor regeneration. Thus, screening of naturally occurring xylose-utilizing micro-organisms is a viable and effective mean to obtain xylitol producing organisms with industrial application. Moreover, the research on selected strains will contribute to a better understanding of regulatory properties of xylose metabolism in different yeasts.

Bioreactors↗

Electrolytic degradation of biorefractory organics and ammonia in leachate from bioreactor landfill.

Electrochemical oxidation was applied to treat the effluent from bioreactor landfill with leachate recirculation, characterised as poor biodegradability and high NH3-N concentration. In this study, the effluent was electrolysed in a batch reactor with Ti/TiO2-IrO2-RuO2 anode and stainless steel cathode. The oxidation of dissolved organic matter (DOM) during electrolysis was evaluated based on the evolution of molecular weight grade, hydrophilic fractionation (humic acid, fulvic acid and hydrophilic fractions), specific ultraviolet absorbance (SUVA254) and AOX. The impact of the initial NH3-N concentration on the oxidation was discussed. The results showed that at a current density of 100 mA/cm2, electrolysis time of 1.5 h and electrode gap of 1 cm, NH3-N with an initial concentration of 1.2 g/L could be completely eliminated and 56% of COD with an initial concentration of 1.2 g/L could be removed, which illustrated that the electrolysis-produced chlorine preferentially oxidised ammonia. The electrolysis mainly resulted in the degradation of humic substances and other high molecular DOM, followed by the increase of BOD/COD ratio and decline of SUVA254 of the leachate. The current efficiencies for COD and ammonia oxidation gradually decreased during the electrolysis, with the latter obviously higher than the former. At the optimal electrolysis time of 1.5 h, NH3-N could be totally removed and the BOD/COD ratio could be enhanced to 0.3, which was also favourable to control the AOX at a reasonable level.

Ammonia↗

Optimum control of the Hemopump as a left-ventricular assist device.

A general framework for designing an optimum control strategy for the Hemopump is described. An objective function was defined that includes four membership functions, each constructed based on the desired values of one of the four members: stroke volume, mean left atrial pressure, aortic diastolic pressure and mean pump rotation speed. The Hemopump was allowed to operate either at a constant speed or at two different speeds during a cardiac cycle. The goal was to maximise the objective function by varying the magnitude and timing of the pump speed. Using a canine circulatory model, it was demonstrated that, in general, different cardiac conditions or different clinical objectives require different operation parameters. For example, when a left ventricle with minor ischaemia was simulated, and the main objective was to increase stoke volume, the objective function was maximised, from a value of 0.877 when the pump was off, to 0.946 when the pump was operated at speed 2 (18 500 revolutions min(-1)). On the other hand, for a severely ischaemic heart, the optimum pump speed became speed 3 (20 000 revolutions min(-1)), which maximized the objective function to 0.943 (from 0.707 when the pump was off). The results also suggest that it is more beneficial to operate the Hemopump at two different speeds during a cardiac cycle (a higher speed during systole and early diastole, and a lower speed during late diastole) than to maintain a constant speed throughout the cardiac cycle.

Animals↗

Conjugation metabolism of acetaminophen and bilirubin in extrahepatic tissues of rats.

An anhepatic rat model was used to explore the extrahepatic conjugating metabolism of acetaminophen and serum bilirubin. The recovery of glucuronide- and sulfate-acetaminophen was 47.5% in normal control and 13.4% in model rats in the urine collected for 6 h after administration of acetaminophen 20 mg kg(-1). Following the increase of acetaminophen dose to 150 mg kg(-1), the recovery of urinary glucuronide-acetaminophen increased by 53.9% in normal control; but it decreased by 36.4% in model rats. In contrast to normal control, the pretreatment with phenobarbital did not affect acetaminophen and its metabolite levels in plasma and urine in model rats. After the establishment of anhepatic model the serum direct bilirubin rose dramatically. Urinary bilirubin test was positive in model rats, but not in normal control. No changes were observed in serum total bilirubin and ratio of direct/total bilirubin after the pretreatment with ranitidine or phenobarbital 50 mg kg (-1), i.p. for 5 days in model rats. The results indicate that the glucuronide- and sulfate-acetaminophen formed in the extrahepatic tissues of model rats is 28.2% of normal control, serum free bilirubin can be transformed into conjugated bilirubin in extrahepatic tissues, and the regulation mechanism of phase II conjugating enzymes is different between the hepatic and extrahepatic tissues.

Acetaminophen↗

Removal of ocular artifacts from electro-encephalogram by adaptive filtering.

The electro-encephalogram (EEG) is useful for clinical diagnosis and in biomedical research. EEG signals, however, especially those recorded from frontal channels, often contain strong electro-oculogram (EOG) artifacts produced by eye movements. Existing regression-based methods for removing EOG artifacts require various procedures for preprocessing and calibration that are inconvenient and time-consuming. The paper describes a method for removing ocular artifacts based on adaptive filtering. The method uses separately recorded vertical EOG and horizontal EOG signals as two reference inputs. Each reference input is first processed by a finite impulse response filter of length M (M = 3 in this application) and then subtracted from the original EEG. The method is implemented by a recursive least-squares algorithm that includes a forgetting factor (lambda = 0.9999 in this application) to track the non-stationary portion of the EOG signals. Results from experimental data demonstrate that the method is easy to implement and stable, converges fast and is suitable for on-line removal of EOG artifacts. The first three coefficients (up to M = 3) were significantly larger than any remaining coefficients.

Algorithms↗

Condition, prolonged swimming performance and muscle metabolic capacities of cod Gadus morhua.

This study evaluated the link between swimming endurance and condition of Atlantic cod Gadus morhua that had been fed or starved during the 16 weeks preceding the tests, and assessed whether muscle metabolic capacities explain such links. The condition factor [(somatic mass x fork length(-3))x100] of starved cod was 0.54+/-0.1 whereas that of fed cod was 0.81+/-0.1. In white and red muscle, we measured four glycolytic enzymes: phosphofructokinase (PFK), pyruvate kinase (PK), creatine kinase (CK) and lactate dehydrogenase (LDH), two mitochondrial enzymes: cytochrome c oxidase (CCO) and citrate synthase (CS), a biosynthetic enzyme, nucleoside diphosphate kinase (NDPK), glycogen and protein levels and water content. Muscle samples were taken at three positions along the length of the fish; starvation affected the metabolic capacities of white muscle more than those of red muscle. The levels of glycolytic enzymes and glycogen changed more in white than red muscle during starvation. Both in fed and starved cod, muscle metabolic capacities varied with position along the fish; starvation reduced this longitudinal variation more in white than red muscle. In white muscle of fed cod, the glycolytic enzyme levels increased from head to tail, while in starved cod this longitudinal variation disappeared. In red muscle mitochondrial enzyme levels were highest in the caudal sample, but fewer differences were found for glycolytic enzymes. Swimming endurance was markedly affected by fish condition, with starved fish swimming only 30% of the time (and distance) of fed fish. This endurance was closely linked with the number of burst-coast movements during the test and the activity of CCO and LDH in white muscle. The number of burst-coast movements was significantly linked with condition factor and PFK activity in caudal red muscle and gill arch mass. Our data indicated that cod use both glycolytic and oxidative capacities to support endurance swimming. Furthermore, swimming endurance is linked with the metabolic capacities of red and white muscle.

Animals↗

Simulation study of the Hemopump as a cardiac assist device.

A dynamic model was developed for a Hemopump that withdraws blood from the left ventricle and discharges it to the aorta through a miniature axial-flow pump. Incorporation of the Hemopump model in a previously established model for the canine circulatory system enabled the effects of the Hemopump on various haemodynamic variables of the circulatory system to be studied, and the benefit of the Hemopump to the failing heart was investigated. In addition, the influence of the physiological status of the right ventricle on the Hemopump performances was examined, and the synchronous and non-synchronous operations of the Hemopump were compared. Results verified that the Hemopump assists the failing heart by increasing the oxygen supply, while reducing the oxygen consumption of the heart through a reduction in the workload of the left ventricle. These beneficial effects were enhanced when the pump's rotation speed was increased. When pump speed was increased from 17,000 to 23,000 revolutions min-1, the oxygen supply increased 101%, and the oxygen consumption decreased 60%. However, when the pump rotation speed was too high, the inflow to the pump could be impaired and the pump performance could be negatively affected. Predications from the model were in good agreement with the results previously obtained in animal experiments and in vitro measurements.

Animals↗

Novel ultrasonic fusion imaging method based on cyclic variation in myocardial backscatter.

Quantitative ultrasonic tissue characterisation of the myocardium based on integrated backscatter (IB) has the potential of becoming an effective method for detecting and evaluating myocardial ischaemia. To facilitate IB-based clinical applications, a new imaging method has been developed that combines the anatomical information of a B-mode image with the contractile performance of a selected myocardial region. To produce such a fusion image, a region of interest (ROI) in a B-mode cardiac image was first selected by the user. Algorithms for detection of the endocardium and epicardium were developed, and the resulting mean distance between the computer-detected curve and the manually traced curve was 0.83mm for the endocardium and 0.58mm for the epicardium. The cyclic variation of IB (CVIB) of each myocardial tissue element within the ROI was then calculated over one cardiac cycle. Finally, a grey-scale B-mode image at the end of diastole was displayed as a still image, and the pixels representing the myocardial tissue in the ROI colour-coded according to the corresponding CVIB over the past heart cycle. Both the B-mode image and the colour-coded region were refreshed (up-dated) at the next end-of-diastole. Preliminary results from normal (CVIB= 10-12dB) and ischaemic (CVIB = 5-7 dB) canine hearts are presented that demonstrate the utility of this new imaging method.

Diagnosis, Computer-Assisted↗

Screening of pyruvate-producing yeast and effect of nutritional conditions on pyruvate production.

AIMS: To find a yeast strain that can overproduce pyruvate and to investigate the effect of nutrients on pyruvate production. METHODS AND RESULTS: Trichosporon cutaneum PD70, a yeast strain that can overproduce pyruvate, was isolated from shake-flask cultures of 132 yeast strains. Pyruvate was measured by the HPLC or DNP method (see Materials and methods). Pyruvate production reached approximately 30.0 +/- 1.0 g l(-1) in basal fermentation medium. Different nutrient supplements had great effects on pyruvate production. Some of the conditions that gave the highest yield are described. CONCLUSIONS: Exogenous thiamine supplement caused a decrease in pyruvate yield. Some amino acids, such as L-arginine, L-isoleucine and L-valine, caused a minor increase in pyruvate yield. Soybean peptone was the most suitable nitrogen source for pyruvate production. A glucose concentration of 15% in fermentation medium gave the highest yield (34.6 g l(-1)) and the highest yield against consumed glucose (0.429 g g(-1)). SIGNIFICANCE AND IMPACT OF THE STUDY: Nutrients have significant impacts on pyruvate production. As a pyruvate overproducing yeast strain independent of exogenous vitamins or amino acids, T. cutaneum PD70 provides an advantage for commercial pyruvate production.

Amino Acids↗

The effects of catechol-O-methyltransferase inhibition on estrogen metabolite and oxidative DNA damage levels in estradiol-treated MCF-7 cells.

Many of the major identified risk factors for breast cancer are associated with exposure to endogenous estrogen. In addition to the effects of estrogen as a growth factor, experimental and epidemiological evidence suggest that catechol metabolites of estrogen also contribute to estrogen carcinogenesis by both direct and indirect genotoxic mechanisms. O-Methylation catalyzed by catechol-O-methyltransferase (COMT) is a Phase II metabolic inactivation pathway for catechol estrogens. We and others have found that a polymorphism in the COMT gene, which codes for a low activity variant of the COMT enzyme, is associated with an increased risk of developing breast cancer; therefore, the goal of the current study was to investigate the role of decreased COMT activity on estrogen catechol levels and on oxidative DNA damage, as measured by 8-hydroxy-2'-deoxyguanosine (8-oxo-dG) levels. MCF-7 cells were pretreated with dioxin as a means to increase estrogen metabolism to catechol estrogens, then treated with estradiol (E2) +/- Ro 41-0960, a COMT-specific inhibitor. After extraction from culture medium, estrogen metabolites were separated using an high-performance liquid chromatography-electrochemical detection method. As expected, dioxin dramatically increased E2 oxidative metabolism, primarily to its 2-OH and 2-methoxy metabolites. The COMT inhibitor blocked 2-methoxy E2 formation. This was associated with increased 2-hydroxy E2 (2-OH E2) and 8-oxo-dG levels. In the presence of COMT inhibition, increased oxidative DNA damage was detected in MCF-7 cells exposed to as low as 0.1 microM E2, whereas in the absence of COMT inhibition, no increase in 8-oxo-dG was detected at E2 concentrations < or =10 microM. This study is the first to show that O-methylation of 2-OH E2 by COMT is protective against oxidative DNA damage caused by 2-OH E2, a major oxidative metabolite of E2.

8-Hydroxy-2'-Deoxyguanosine↗

First staging of two laser accelerators.

Staging of two laser-driven, relativistic electron accelerators has been demonstrated for the first time in a proof-of-principle experiment, whereby two distinct and serial laser accelerators acted on an electron beam in a coherently cumulative manner. Output from a CO2 laser was split into two beams to drive two inverse free electron lasers (IFEL) separated by 2.3 m. The first IFEL served to bunch the electrons into approximately 3 fs microbunches, which were rephased with the laser wave in the second IFEL. This represents a crucial step towards the development of practical laser-driven electron accelerators.

Journal Article↗

[A study of the hereditary susceptibility of HLA-DQA1 to essential hypertension, athrothrombotic brain infarction and lacunar stroke].

OBJECTIVE: To study the types of HLA-DQA1 alleles in patients with essential hypertension (EH), athrothrombotic brain infarction (ABI) and lacunar stroke (LS) and normal control individuals and the hereditary susceptibility of HLA-DQA1 alleles to those diseases. METHODS: The allelic typing of HLA-DQA1 was detected by PCR-SSP in 155 cases of EH, ABI and LS and 64 normal individuals. RESULTS: The frequencies of HLA-DQA1 * 0301 in EH, ABI and LS groups were obviously higher than that in normal control group (33.6538 vs. 17.9688, 36.5884 vs. 17.9688, 33.0645 vs. 17.9688, P < 0.01). The frequencies of HLA-DQA1 * 0103 in groups of EH, ABI, LS, ABI with EH, and LS with EH were lower than that in the normal control group (6.7308 vs17.1875, RR = 0.3916, P < 0.01; 6.0976 vs. 17.1875, RR = 0.3548, P < 0.05; 6.4566 vs17.1875, RR = 0.3754, P < 0.01; 2.3809 vs17.1875, RR = 0.1385, P < 0.01; 3.5714 vs. 17.1875, RR = 0.2078, P < 0.01. CONCLUSION: HLA-DQA1 * 0301 allele may be a correlative gene with hereditary susceptibility of EH, ABI and LS, and HLA-DQA1 * 0103 allele may be a protective gene of those diseases.

Adult↗

Characterization of single-stranded DNA on chitosan-modified electrode and its application to the sequence-specific DNA detection.

Single-strand DNA could bind with chitosan on a platinum electrode via forming a tight DNA-chitosan complex. The salt concentration of the ssDNA solution had an obvious effect on the surface coverage, the immobilization was remarkably reduced at high salt concentration. The sample ssDNA immobilized on the chitosan-modified electrode can hybridize efficiently with the complementary sequences and be successfully used for the sequence-specific DNA detection. The same results could be obtained using a gold or graphite electrode modified with chitosan. The stability of this electrode has been also discussed.

Biopolymers↗

Gaf-1, a gamma -SNAP-binding protein associated with the mitochondria.

The role of alpha/beta-SNAP (Soluble NSF Attachment Protein) in vesicular trafficking is well established; however, the function of the ubiquitously expressed gamma-SNAP remains unclear. To further characterize the cellular role of this enigmatic protein, a two-hybrid screen was used to identify new, gamma-SNAP-binding proteins and to uncover potentially novel functions for gamma-SNAP. One such SNAP-binding protein, termed Gaf-1 (gamma-SNAP associate factor-1) specifically binds gamma- but not alpha-SNAP. The full-length Gaf-1 (75 kDa) is ubiquitously expressed and is found stoichiometrically associated with gamma-SNAP in cellular extracts. This binding is distinct from other SNAP interactions since no alpha-SNAP or NSF coprecipitated with Gaf-1. Subcellular fractionation and immunofluorescence analysis show that Gaf-1 is peripherally associated with the outer mitochondrial membrane. Only a fraction of gamma-SNAP was mitochondrial with the balance being either cytosolic or associated with other membrane fractions. GFP-gamma-SNAP and the C-terminal domain of Gaf-1 both show a reticular distribution in HEK-293 cells. This reticular structure colocalizes with Gaf-1 and mitochondria as well as with microtubules but not with other cytoskeletal elements. These data identify a class of gamma-SNAP interactions that is distinct from other members of the SNAP family and point to a potential role for gamma-SNAP in mitochondrial dynamics.

Adaptor Proteins, Signal Transducing↗

Immunophenotyping of lymphocyte T and B in the peripheral blood of systemic lupus erythematosus.

The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus (SLE) were analyzed by using the immunofluorescence labeling-flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD3+, CD4+ and the ratio of CD4+/CD8+ in the peripheral blood of these patients were decreased (P < 0.01), while the expression of CD8+, CD20+ was significantly increased (P < 0.01). It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.

Adolescent↗