Biomedical subjects

L Du

Publications and source records attributed to L Du.

At least 37 records · Page 2Linked to original sources

PubMed2001

Biosynthesis of hybrid peptide-polyketide natural products.

The structural and catalytic similarities between non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) support the idea of combining individual NRPS and PKS modules for combinatorial biosynthesis. Recent advances in cloning and characterization of biosynthetic gene clusters for naturally occurring hybrid polyketide-peptide metabolites have provided direct evidence for the existence of hybrid NRPS-PKS systems, thus setting the stage to investigate the molecular basis for intermodular communication between NRPS and PKS modules. Reviewed in this article are biosynthetic data pertinent to hybrid peptide-polyketide biosynthesis published up to late 2000. Hybrid peptide-polyketide natural products can be divided into two classes: (i) those whose biosyntheses do not involve functional interaction between NRPS and PKS modules; and (ii) those whose biosyntheses are catalyzed by hybrid NRPS-PKS systems involving direct interactions between NRPS and PKS modules. It is the latter systems that are most likely amenable to combinatorial biosynthesis. The same catalytic sites appear to be conserved in both hybrid NRPS-PKS and normal NRPS or PKS systems, with the exception of the ketoacyl synthase domains in hybrid NRPS-PKS systems which are unique. Specific linkers may play a critical role in communication, facilitating the transfer of the growing intermediates between the interacting NRPS and/or PKS modules. In addition, phosphopantetheinyl transferases with broad carrier protein specificity are essential for the production of functional hybrid NRPS-PKS megasynthetases. These findings should now be taken into consideration in engineered biosynthesis of hybrid peptide-polyketide natural products for drug discovery and development.

Animals↗

PubMed2001

The therapeutic effects of Sheng Jing Zhong Zi Tang in treating male infertility.

Eight-seven cases of male infertility due to spermatopathy were treated with Sheng Jing Zhong Zi Tang (SJZZT [symbol: see text]) for 1-3 courses. The total effective rate was 95.40% (83/87). 56.32% (49/87) of their spouses got pregnant and the semen quality was markedly improved (P < 0.01). The results showed that the decoction could dual-directionally regulate the levels of follicle-stimulating hormone (FSH), prolan B luteinizing hormone (LH), testosterone (T), and cortisol (C).

Adult↗

PubMed2001

[Characteristics of N2O emissions from vegetal soils on Fildes peninsula, Antarctica].

The N2O fluxes from the vegetal soils were first measured on the Fildes peninsula, Antarctica, and the total N2O emission was also estimated in the summer 2 months. The daily variations of N2O fluxes appeared single-peak trend under the sunshine or rainy weather conditions but they were irregular under the snow weather conditions and inconsistent with the atmospheric temperatures. The seasonal variations of the N2O fluxes were affected by the temperature and rainfall. The conditions during the transitions between dry and wet seasons improved the N2O emission. The total N2O emissions from moss and lichen soils were 3.7152 kg and 2.5344 kg, respectively. It follows that the vegetal soils are the sources for the atmospheric N2O on the Fildes peninsula, Antarctica.

Antarctic Regions↗

PubMed2001

[An epidemiological and clinical study of primary gout].

OBJECTIVE: To evaluate the prevalence of primary gout in Shantou and Beijing area, and understand it's clinical features. METHODS: Three epidemiological studies in Chenghai downtown, and the one of study in Beijing were studied. Clinical, laboratory and radiology data were collected and analyzed. RESULT: The prevalence of primary gout in Chenghai surveys was 0.17%, 0.15% and 0.26% respectively; no case of primary gout was found in Beijing survey. Among the patients, male to female ratio was 21:1; average onset age of females was significantly higher than that of males. The numbers of patient in the recent 10 years was increasing. At the first attack, the first metatarsophalangeal joint was involved among 79.7% of all cases; no fourth metatarsophalangeal joint involvement had been found. Hyperlipemia was the most common accompanying abnormal laboratory test (57.1%). Radiographic bone erosion was found in 49.4% patients, most of them, the duration of disease was over 5 years. CONCLUSION: The prevalence of primary gout was higher in Shantou area than that in Beijing, and it was increasing in the recent 10 years in Shantou area. The difference of geography, climate, food and lifestyle could be responsible for the different prevalence in different area.

Adult↗

PubMed2001

[A quantitative pathological study on angiogenesis, vascular endothelial growth factor and inducible nitric oxide synthase in astrocytomas].

OBJECTIVE: To investigate whether the expression of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) are associated with angiogenesis and malignancy of astrocytomas. METHODS: Immunohistochemistry, semiquantitation and image analysis were used to study the expression features of VEGF and iNOS in 61 cases of astrocytomas having follow-up data. Factor VIII related antigen (FVIII RAg) was also analysed in reflecting the epithelial cell amount and the microvessel density. RESULTS: Microvessels in astrocytomas could be divided into seven types. The positive reaction areas and IA values of FVIIIRAg expression in the endothelium were significantly increased with the increase of tumor grade (P < 0.001) and VEGF labeling index (LI) (P < 0.05). The patients with high VEGF LIs (LI >or= 25%) had a poor prognosis. VEGF expression was significantly related to that of iNOS (P < 0.001), i.e., VEGF level was reduced along with the decrease of iNOS reaction. CONCLUSION: Detection of FVIIIRAg could reflect the angiogenesis activities in astrocytomas. VEGF and iNOS could up-regulate each other to promote angiogenesis and are of importance in determining the malignancy degree of astrocytomas.

Astrocytoma↗

PubMed2001

[Studies on correlation between protonation and tautomerism and luminescence characteristics of quinolone].

The fluorescence characteristics of 8 kinds of quinolone in different pH medium were studied. Rules of wavelength and intensity of the fluorescence in different pH medium were discovered. The correlation between the protonation, tautomerism and luminescence characteristics of quinolone are studied. It is found that in pH 6-8 or 2-4 there are two luminescence species. By this, the new view is represented that the change of luminescence characteristics is arised from the protonation and tautomerism of quinolones in different pH medium and the material point of luminescence developed in equilibrium system of tautomerism. Additionally, the correlation between the protonation, tautomerism and luminescence is further discussed.

Anti-Infective Agents↗

PubMed2001

[Determination of ofloxacin in urine by synchronous-derivative fluorimetry].

The internal hormones in human urine emit strong fluorescence and interfere with the determination of trace ofloxacin. However, the experiments indicate that synchronous-first order derivative fluorimetry can be applied to the determination of trace ofloxacin without interference. The linear range is 0.36-3.6 micrograms.mL-1, detection limit is 0.3 microgram.mL-1, RSD is less than 2.5% and the recovery is within the range of 101%-103%.

Humans↗

PubMed2001

[Simultaneous determination of hydride forming and non-hydride forming elements in human hair by hydride generation-inductively coupled plasma atomic emission spectrometry].

A method for simultaneous determination of both hydride forming and non-hydride forming elements in human hair has also been successful. The influence of HCl concentration, NaBH4 concentration, carrier gas flow rate and rinse time on hydride forming elements intensity, and the effect of between nebulization and hydride-nebulization system on non-hydride forming elements have been studied. The optimum experimental conditions are selected. This method has been applied to the analysis of standard human hair with satisfactory results.

Arsenic↗

PubMed2000

Modulation of mitogen-activated protein kinases and phosphorylation of Bcl-2 by vinblastine represent persistent forms of normal fluctuations at G2-M1.

Microtubule inhibitors, widely used in cancer chemotherapy, induce G2-M arrest and apoptosis and have in common the ability to stimulate Raf-1/Bcl-2 phosphorylation and activate c-Jun NH2-terminal protein kinase (JNK). These signal transduction pathways are thought to be activated in response to microtubule damage to promote apoptosis. However, Bcl-2 phosphorylation has been reported to occur at G2-M in nonapoptotic cells, raising the possibility that this and perhaps other signaling pathways altered by microtubule inhibitors reflect perturbations of normal mitotic events. In this study, we sought to test this hypothesis. We show that Bcl-2 phosphorylation and JNK activation, as well as extracellular response kinase and p38 inactivation, occur not only in response to vinblastine but also as discrete transient events at G2-M phase in untreated synchronized KB-3 cells. Thus, modulation of these pathways is not a response to microtubule damage; rather they occur normally at G2-M, and it is the extent, duration, and/or irreversible nature of the signals that distinguish a preapoptotic cell from one destined to divide. These findings provide novel insight into the relationship between mitotic and apoptotic signaling and the mechanism of action of antimitotic drugs.

Activating Transcription Factor 2↗

PubMed2000

An oxidation domain in the BlmIII non-ribosomal peptide synthetase probably catalyzing thiazole formation in the biosynthesis of the anti-tumor drug bleomycin in Streptomyces verticillus ATCC15003.

We have previously proposed that the BlmIV and BlmIII non-ribosomal peptide synthetases are involved in the formation of the bithiazole moiety of the anti-tumor drug bleomycin in Streptomyces verticillus ATCC15003. We report here the identification and characterization of an oxidation domain in BlmIII. The oxidation domain shows local homology to a family of oxidoreductases and is present in all thiazole-forming non-ribosomal peptide synthetase modules known to date. Both the blmIII-Ox domain and blmIII gene were expressed in Escherichia coli, and the resulting BlmIII-Ox and BlmIII proteins were purified to homogeneity. The oxidation domain contains one molar equivalent of non-covalently bound FMN as a prosthetic group. These results provide experimental evidence for an oxidation domain within non-ribosomal peptide synthetases, suggesting that BlmIII-Ox probably catalyzes the thiazoline to thiazole oxidation in bleomycin biosynthesis.

Amino Acid Sequence↗

PubMed2000

During apoptosis of HL-60 and U-937 cells caspases are activated independently of dissipation of mitochondrial electrochemical potential.

Collapse of the mitochondrial potential (DeltaPsi(m)) during apoptosis has been linked with a release of cytochrome c and apoptosis-inducing factor (AIF) and activation of caspases. Using a laser scanning cytometer (LSC), an instrument that allows one to measure the same cells twice, first when they are alive and subsequently after their permeabilization, we explored whether dissipation of DeltaPsi(m) (measured supravitally) is a prerequisite for the activation of caspases (detected after cell fixation). Apoptosis of HL-60 cells was induced either by TNF-alpha combined with cycloheximide (CHX) or by the DNA topoisomerase I inhibitor camptothecin (CPT) and of U-937 cells by CPT, and DeltaPsi(m) was measured using the carbocyanine fluorochrome DiIC(1) (5). The marker of caspase activation was specific cleavage of poly(ADP) ribose polymerase (PARP) detected immunocytochemically. After 30 or 60 min treatment with TNF-alpha + CHX or 60 or 120 min with CPT a considerable proportion of cells (20-40%) demonstrated PARP cleavage with no evidence of DeltaPsi(m) collapse. Also present in these cultures (3-20%) were cells with collapsed DeltaPsi(m) whose PARP was not cleaved. The results provide direct evidence that in HL-60 and U-937 cells treated with TNF-alpha + CHX or CPT the dissipation of DeltaPsi(m) is not required for activation of caspases and these two events are independent of each other.

Apoptosis↗

PubMed2000

Association of polymorphism of serotonin 2A receptor gene with suicidal ideation in major depressive disorder.

There is evidence indicating that density of 5-HT2A receptors is altered in brain regions of depressed suicide victims and in platelets of suicidal subjects with major depression or schizophrenia. Recent studies have also shown an association between the allele C of 102T/C polymorphism in the 5-HT2A receptor gene and schizophrenia. The present investigation tested the hypothesis that the observed changes in 5-HT2A receptor density in platelets of patients with major depression are a trait rather than state phenomenon and are associated with the 102 C allele in 5-HT2A receptor gene in a sample of 120 patients with major depression and a group of 131 control subjects comparable with respect to age, sex, and ethnic background. The allele and genotype frequencies of 102T/C polymorphism in 5-HT2A receptor gene were compared between patients and control subjects and between suicidal and non-suicidal patient groups. The major finding of this study was a significant association between the 102 C allele in 5-HT2A receptor gene and major depression, chi(2) = 4.5, df = 1, P = 0.03, particularly in patients with suicidal ideation, chi(2) = 8.5, df = 1, P < 0.005. Furthermore, we found that patients with a 102 C/C genotype had a significantly higher mean HAMD item 3 score (indication of suicidal ideation) than T/C or T/T genotype patients. Our results suggest that the 102T/C polymorphism in 5-HT2A receptor gene is primarily associated with suicidal ideation in patients with major depression. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:56-60, 2000.

Adult↗

PubMed2000

Reduction by naloxone of lipopolysaccharide-induced neurotoxicity in mouse cortical neuron-glia co-cultures.

An inflammatory response in the CNS mediated by activation of microglia is a key event in the early stages of the development of neurodegenerative diseases. Using mouse cortical mixed glia cultures, we have previously demonstrated that the bacterial endotoxin lipopolysaccharide induces the activation of microglia and the production of proinflammatory factors. Naloxone, an opioid receptor antagonist, inhibits the lipopolysaccharide-induced activation of microglia and the production of proinflammatory factors. Using neuron-glia co-cultures, we extended our study to determine if naloxone has a neuroprotective effect against lipopolysaccharide-induced neuronal damage and analysed the underlying mechanism of action for its potential neuroprotective effect. Pretreatment of cultures with naloxone (1 microM) followed by treatment with lipopolysaccharide significantly inhibited the lipopolysaccharide-induced production of nitric oxide and the release of tumor necrosis factor-alpha, and significantly reduced the lipopolysaccharide-induced damage to neurons. More importantly, both naloxone and its opioid-receptor ineffective enantiomer (+)-naloxone were equally effective in inhibiting the lipopolysaccharide-induced generation of proinflammatory factors and the activation of microglia, as well as in the protection of neurons. These results indicate that the neuroprotective effect of naloxone is mediated by its inhibition of microglial activity and may be unrelated to its binding to the classical opioid receptors.

Animals↗

PubMed2000

The biosynthetic gene cluster for the antitumor drug bleomycin from Streptomyces verticillus ATCC15003 supporting functional interactions between nonribosomal peptide synthetases and a polyketide synthase.

BACKGROUND: The structural and catalytic similarities between modular nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) inspired us to search for a hybrid NRPS-PKS system. The antitumor drug bleomycin (BLM) is a natural hybrid peptide-polyketide metabolite, the biosynthesis of which provides an excellent opportunity to investigate intermodular communication between NRPS and PKS modules. Here, we report the cloning, sequencing, and characterization of the BLM biosynthetic gene cluster from Streptomyces verticillus ATCC15003. RESULTS: A set of 30 genes clustered with the previously characterized blmAB resistance genes were defined by sequencing a 85-kb contiguous region of DNA from S. verticillus ATCC15003. The sequenced gene cluster consists of 10 NRPS genes encoding nine NRPS modules, a PKS gene encoding one PKS module, five sugar biosynthesis genes, as well as genes encoding other biosynthesis, resistance, and regulatory proteins. The substrate specificities of individual NRPS and PKS modules were predicted based on sequence analysis, and the amino acid specificities of two NRPS modules were confirmed biochemically in vitro. The involvement of the cloned genes in BLM biosynthesis was demonstrated by bioconversion of the BLM aglycones into BLMs in Streptomyces lividans expressing a part of the gene cluster. CONCLUSION: The blm gene cluster is characterized by a hybrid NRPS-PKS system, supporting the wisdom of combining individual NRPS and PKS modules for combinatorial biosynthesis. The availability of the blm gene cluster has set the stage for engineering novel BLM analogs by genetic manipulation of genes governing BLM biosynthesis and for investigating the molecular basis for intermodular communication between NRPS and PKS in the biosynthesis of hybrid peptide-polyketide metabolites.

Amino Acid Sequence↗

PubMed2000

Adherence of Porphyromonas gingivalis to gingival epithelial cells: modulation of bacterial protein expression.

The protein profiles of Porphyromonas gingivalis (ATCC 33277 and W83) bound to KB gingival epithelial cells were analyzed by SDS-PAGE and immunoblotting. We found that a 51-kDa component was formed in bacteria that adhered to the KB cells, whereas 26- to 29-kDa bands were less intensive, in contrast to the protein profile of free bacteria. P. gingivalis ATCC 33277 incubated with protease-treated KB cells retained the profile of free bacteria. These results demonstrate the specificity of bacterial recognition of eukaryotic membrane components.

Adhesins, Bacterial↗

PubMed2000

Identification of genes encoding receptor-like protein kinases as possible targets of pathogen- and salicylic acid-induced WRKY DNA-binding proteins in Arabidopsis.

To understand how plant host genes are regulated during the activation of plant defence responses, we are studying a group of pathogen- and salicylic acid (SA)-induced DNA-binding proteins containing the novel WRKY domain. To identify downstream target genes of these WRKY proteins, we have searched the Arabidopsis genome and identified four closely linked genes on chromosome IV that contain an unusually large number of the W-box sequences [(T)TGAC(C/T)] recognized by WRKY proteins within a few hundred base pairs upstream of their coding regions. All four genes encode proteins characteristic of receptor-like protein kinases (RLK), each consisting of an N-terminal signal sequence, an extracellular receptor domain, a single transmembrane domain and a C-terminal cytoplasmic serine/threonine protein kinase domain. All four RLK genes were induced by treatment with SA or infection by a bacterial pathogen. Studies with one of the RLK genes (RLK4) indicated that a cluster of W-box elements in its promoter region were recognized by both purified WRKY proteins and SA-induced W-box binding activities from SA-treated Arabidopsis plants. Further analysis using the RLK4 gene promoter fused to a reporter gene in transgenic Arabidopsis indicated that the consensus WRKY protein-binding sites in the RLK4 gene promoter were important for the inducible expression of the reporter gene. These results indicate that pathogen- and SA-induced W-box binding proteins regulate not only genes encoding defence proteins with direct or indirect anti-microbial activities, but also genes encoding proteins with regulatory functions.

Amino Acid Sequence↗

PubMed2000

Gender differences in association between serotonin transporter gene polymorphism and personality traits.

Since Lesch and colleagues reported an association between anxiety-related traits (Neuroticism) and a functional polymorphism in the serotonin transporter gene regulatory region (5-HTTLPR), there have been several reports on 5-HTTLPR and personality traits with both positive and negative results. The present study was a further attempt to replicate the original findings of Lesch et al. in a population of well-defined normal healthy subjects. In addition, a variable number tandem repeat polymorphism in the second intron was included in this study because it has recently been shown to act as a transcriptional regulator. Personality traits were evaluated in 186 unrelated normal subjects by the NEO Five Factor Inventory. The most important and novel finding of this study was a significant association of mean Neuroticism scores with the short allele of 5-HTTLPR in male subjects (t = 2.4, P = 0.018). We were thus able to replicate the finding of Lesch et al. of an association between serotonin transporter gene polymorphism (5-HTTLPR) and Neuroticism, but only in a male population. We also found a significant effect of gender on mean scores of Neuroticism [F = 3.9, degrees of freedom (df) = 1, 180, P = 0.05] and Agreeableness (F = 6.8, df = 1, 180, P = 0.01), but no significant effect of 5-HTTLPR genotype on Neuroticism (F = 0.87, df= 2, 180, P = 0.42) or Agreeableness (F = 0.35, df = 2, 180, P = 0.7). These findings suggest that gender differences exist in contribution of genetic factors to behavioural phenotypes. They may also explain the inconsistencies in previous reports on association of Neuroticism with 5-HTTLPR from studies using different proportions of male and female subjects.

Adult↗

PubMed2000

Modeling a protein foam fractionation process.

A simple staged model for the protein foam fractionation process is proposed in this article. This simplified model does not detail the complex foam structure and gas-liquid hydrodynamics in the foam phase but, rather, is built on the conventional theoretical stage concept considering upward bubbles with entrained liquid and downward liquid (drainage) as counter-current flows. To simulate the protein concentration distribution in the liquid along the column by the model, the bubble size and liquid hold-up with respect to the position must be known, as well as the adsorption isotherm of the protein being considered. The model is evaluated for one stage by data from the semibatch foam fractionation of egg albumin and data from the continuous foam fractionation of bovine serum albumin. The effect of two significant variables (superficial gas velocity and feed protein concentration) on enrichment is well predicted by the model, especially for continuous operation and semibatch operation when initial concentration is high.

Adsorption↗