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D Wu

Publications and source records attributed to D Wu.

At least 361 records · Page 20Linked to original sources
PubMed1997

Dynorphin A1-13 stimulates ovine fetal pituitary-adrenal function through a novel nonopioid mechanism.

We previously reported that U50488H, a kappa-selective opioid agonist, stimulates the release of adrenocorticotropin (ACTH) in the ovine fetus via the release of hypothalamic arginine vasopressin and corticotropin releasing factor. In this study we examined the effects of the endogenous kappa-preferring opioid peptide, dynorphin A1-13, on fetal ACTH release using the unanesthetized, chronically catheterized fetal lamb model. Fetal plasma samples were collected at timed intervals after fetal administration of dynorphin A1-13 (0.5 mg/kg, i.v.) and subsequently analyzed by radioimmunoassay for immunoreactive-ACTH and immunoreactive-cortisol. Dynorphin A1-13 produced a highly significant and rapid increase in immunoreactive-ACTH (P = .002) and immunoreactive-cortisol (P = .002) with peak levels of 383.3 +/- 43.8 pg/ml and 32.8 +/- 9.0 ng/ml, respectively, at 15 min after administration. A similar increase in plasma immunoreactive-ACTH was seen after the same dose of dynorphin A1-17 (P = .02) but not dynorphin A2-17. This ACTH response to dynorphin A1-13 was shown to be insensitive to the opioid antagonist, naloxone (12 mg/hr), as well as antagonists of corticotropin releasing factor and arginine vasopressin. These data suggest that dynorphin A1-13 in the ovine fetus may be acting through a mechanism distinct from the kappa-opioid system and that the dynorphins may serve as secretagogues of ACTH directly at the anterior pituitary through nonopioid receptors.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1997

Fluorine-18-FPCT: a PET radiotracer for imaging dopamine transporters.

UNLABELLED: Fluorine-18-labeled 2 beta-carbomethoxy-3 beta-(4-chlorophenyl)-8-[-3-fluoropropyl) nortropane (FPCT) has been synthesized as a new dopamine transporter imaging agent. METHODS: Fluorine-18 was introduced into 2 beta-carbomethoxy-3 beta-(4-chlorophenyl)-8-[-3-fluoropropyl) nortropane by preparation of 1-[18F]fluoro-3-iodopropane followed by alkylation of 2 beta-carbomethoxy-3 beta-(4-chlorophenyl)nortropane. RESULTS: Tissue distribution studies in rats with [18F]FPCT showed high striatal uptake (0.70% dose/g at 60 min; 0.38% dose/g at 120 min) and good striatal-to-cerebellum ratios (5.5 at 60 min; 6.2 at 120 min). Imaging studies in rhesus monkeys (n = 2) with [18F]FPCT showed high uptake and retention in the putamen (P) (P = 0.03%-0.12% dose/g; at 115 min) and good putamen-to-cerebellum ratios of 3.40-3.43 at 115 min. Plasma metabolites were analyzed in rhesus monkeys (n = 2) by ether extraction and HPLC. The radioactivity in the ether-extractable fraction displayed a single peak that corresponded on HPLC to unmetabolized authentic FPCT. CONCLUSION: These results suggest that [18F]FPCT is an excellent candidate for PET imaging of dopamine transporters.

Animals↗
PubMed1997

Opioid modulation of the fetal hypothalamic-pituitary-adrenal axis: the role of receptor subtypes and route of administration.

The role of receptor subtypes in opioid modulation of the hypothalamic-pituitary-adrenal (HPA) axis is well understood in the adult but has not been investigated in the developing fetus. Because the fetal HPA axis plays an important role in the development of several vital organs and in the onset of parturition, an understanding of the role of opioid receptor subtypes on the fetal HPA axis is important in the design of new obstetrical analgesics. In these studies, we examined the effects of highly selective mu, delta and kappa opioid agonists on plasma immunoreactive adrenocorticotropin (ir-ACTH) and immunoreactive cortisol (ir-cortisol) in the ovine fetus. Intravenous administration of the mu selective agonist [D-Ala2-N-Me-Phe4,Gly-ol]-enkephalin resulted in a 92% increase in ir-ACTH (P = .005) and ir-cortisol. The delta selective agonist, [D-Pen2,D-Pen5]-enkephalin, elicited a much smaller increase (52%) in ir-ACTH (P = .01). In contrast, there was a 7-fold increase in ir-ACTH (P < .001) and a significant increase in ir-cortisol (P = .02) with the kappa selective U50,488H. When the same agonists were administered intracerebroventricularly, there was no change in ir-ACTH or ir-cortisol. These data suggest that the kappa opioid receptor may be more important in the modulation of the fetal HPA axis and that the distribution of these opioid agonists from the lateral ventricle to the hypothalamus and pituitary is very limited.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1997

Analysis of epitope structure of PSP94 (prostate secretory protein of 94 amino acids): (I). Immuno-dominant and immuno-recessive area.

PSP94 is a potential biomarker for evaluating patients with prostate carcinoma. We have systematically studied the epitope structure of PSP94 by using a polyclonal antibody against human PSP94. Results of peptide mapping and ELISA tests of dose response to rabbit antiserum against human PSP94 protein showed that only the N-terminal peptides (N30 and M23) are immunoreactive while all the synthetic peptides (C28, C10) located closer to the C-terminus are completely devoid of antigenic activity with the polyclonal antibody. These results were confirmed by analysis of reciprocal competitive binding of PSP94 polyclonal antibody by the N-terminal peptides (N30 and M23) v. either recombinant GST-PSP94 fusion protein, purified recombinant PSP94, or natural PSP94 protein. To further delineate the antigenic activity of the N- and C-termini, we have also expressed N- and C-terminal half of the whole PSP94 (each 47 peptides) using the E. coli GST expression system. The recombinant N47/C47 peptides were released by thrombin cleavage from the GST fusion protein and characterized by Western blotting experiments. Dose response of the recombinant GST-PSP-N47 and -C47 peptides to PSP94 polyclonal antibody showed differential binding activities. Competitive binding of these recombinant N47/C47 proteins against the GST-PSP94 protein demonstrates that the polyclonal antibody has a higher affinity for the N47 peptide than the C47 peptide. Based on the immunological studies of both synthetic peptides and recombinant PSP94- N/C terminal proteins, we propose an epitope structure of human PSP94 with an immuno-dominant N-terminus and an immuno-recessive C-terminus.

Amino Acid Sequence↗
PubMed1997

Analysis of epitope structure of PSP94 (prostate secretory protein of 94 amino acids): (II). Epitope mapping by monoclonal antibodies.

PSP94 has shown potential to be a serum biomarker for evaluating prostate cancer. Studies of the epitope structure is crucial for this endeavour. In this article, we have used 15 different monoclonal antibodies (MAb) to analyse the epitope structure of PSP94 and to compare with the results obtained from our previous work using polyclonal antibody and recombinant PSP94. Firstly, we determined the relative activities of the 15 MAb population by direct and competitive ELISA. The two predominant MAbs (MAb PSP-6 and -19) in 15 MAbs were selected for further studies of the epitope structure. By comparing the binding activities of recombinant GST-PSP94 and natural PSP94 with MAbs, and by comparing their affinity with MAbs in an in vitro denaturing experiment, PSP94 was shown to have a similar, prevalently linear epitope structure as we demonstrated by polyclonal antibody. Using recombinant GST fusion protein with PSP94 and with each half of the N- and C-terminal 47 amino acids (GST-PSP-N47/C47) in E. coli cells, the different epitopes recognized by 15 monoclonal antibodies were delineated and the polar distribution of the epitope structure of PSP94 was characterized. Results of direct ELISA of recombinant N47 and C47 and their competitive binding against natural PSP94 (competitive ELISA) showed that the N- and C-termini represent the immuno-dominant and immuno-recessive area separately. A majority of the monoclonal antibodies (12/15) showed preferential binding of the N-terminal sequence of the PSP94 protein. Using GST-PSP-N47 as a standard protein, an epitope map of the 15 monoclonal antibodies was obtained. The results of this study will help to define the clinical utility of PSP94.

Animals↗
PubMed1997

Blood-brain barrier permeability to morphine-6-glucuronide is markedly reduced compared with morphine.

The blood-brain barrier (BBB) permeability to morphine and morphine-6-glucuronide (M6G) is measured under identical conditions using an intravenous injection method in the rat and HPLC separation of morphine from its metabolites. The brain uptake of M6G expressed as %ID/g was 32-fold lower than that of morphine, and the BBB permeability surface area product (PS) of M6G was 57-fold lower as compared with that of morphine. Consistent with these in vivo data, the 1-octanol/buffer partition study showed the liposolubility of M6G was 187-fold lower than that of morphine. The CNS origin of M6G analgesia after peripheral administration was confirmed because the analgesia was completely blocked by naloxone, which crosses BBB, but not by naloxone methiodide, which does not enter brain from blood. In conclusion, the BBB permeability to M6G is markedly reduced as compared with morphine, consistent with the much lower lipid solubility of M6G relative to morphine.

Animals↗
PubMed1997

Comparison of clinical efficacy and adverse effects between extended-release felodipine and atenolol in patients with mild and moderate essential hypertension.

BACKGROUND: Essential hypertension is a risk factor for cardiovascular disease. Atenolol, a cardio-selective beta-blocker, has been shown to be a safe and effective antihypertensive agent. The extended-release form of felodipine (felodipine ER), a vascular-selective dihydropyridine calcium blocker, is extensively used in Caucasians. However, its effectiveness, tolerability and adverse side-effect have not been assessed in Chinese populations. METHODS: Sitting blood pressure (BP), heart rate, body weight, adverse reaction and serum biochemistry were assessed in 70 patients with mild-moderate essential hypertension treated either with felodipine ER (37 patients), or atenolol (33 patients) for 10 weeks. Each patient was prescribed 5 mg of felodipine ER or 50 mg of atenolol once daily and this daily dosage was doubled to twice daily if necessary. RESULTS: Six patients who received felodipine ER and 3 who received atenolol withdrew from the treatment because of intolerable side effects. Within ten weeks, 81.1% of the patients had responded to a total daily dosage of 5-10 mg of felodipine ER and 81.8% to a daily dose of 50-100 mg of atenolol. By the end of treatment, the mean BP in the felodipine ER group had decreased from 176/104 mmHg at baseline to 145/85 mmHg, while the BP in the atenolol group had dropped from 173/103 mmHg to 145/84 mmHg (NS between the two groups). Heart rate declined in the atenolol group but did not change in patients who received felodipine ER. Overall, patients in the felodipine ER group had a higher rate of adverse reaction (70.3% vs. 39.4%; p < 0.001), and 16.2% of the patients in the felodipine ER group experienced symptoms of hypotension. CONCLUSION: Equivalent doses of felodipine ER and atenolol are effective first-line monotherapeutic agents for the treatment of mild-moderate essential hypertension.

Adrenergic beta-Antagonists↗
PubMed1997

Effect of hydroxyl radical scavenging capacity on clustering of DNA damage.

We have shown previously that the thiol N-(2'-mercaptoethyl)-1,3-diaminopropane (WR-1065) can attenuate the formation of strand breaks associated with ionizing radiation. The mechanism of this protection is predominantly the reduction of DNA radical species which otherwise would attenuate the chemical repair of DNA radical species which are strand break precursors. We had observed that the presence of a hydroxyl radical scavenger during irradiation resulted in a decrease in the ability of WR-1065 to attenuate the formation of strand breaks. Since ionic compounds are known to affect the binding of the dicationic WR-1065 with the polyanion DNA, the effect of the scavenger was initially attributed to its polar nature having a similar effect on the interaction of WR-1065 with DNA, and not as a consequence of its ability to scavenge hydroxyl radicals. After examining additional scavengers, we now conclude that an increased hydroxyl radical scavenging capacity does attenuate the repair of strand break precursors to some extent. The probable explanation for this observation is that an increased scavenging capacity results in a greater degree of radical clustering on the DNA, and that these clusters of multiple radicals are repaired more slowly than are single radical species.

DNA Damage↗
PubMed1997

Body surface Laplacian mapping of bioelectrical activity.

A method is described to process and interpret multi-channel bioelectrical signals. The bioelectrical signals were recorded noninvasively over the body surface of human subjects at 120 sites. The body surface Laplacian maps were then constructed from the multi-channel bioelectrical potential measurement. The method was evaluated by means of computer simulations, and applied to imaging cardiac electrical activity. The present investigation suggests body surface Laplacian mapping provides an important means in interpreting bioelectrical signals.

Body Surface Potential Mapping↗
PubMed1997

[Tissue engineering study on repairment of injured nerve gap in rat].

The combined substances composed of chitosan gel and rough abstract contained schwanns cell (RSC) plus extra nerve growth factor (NGF) were filled in silicone chambers to bridge a 10 mm nerve gap in rat. The RSC were created by removing autonerve's epineurium and perineurium. The very same other silicone chambers with only NGF saline solution were used as control. Electrophysiologic recording and transverse section imaging analysis at 12 weeks postoperatively showed better result in the experimental group, compared with the control group. The research suggests a new tissue engineering method for repairing nerve gap.

Animals↗
PubMed1997

Effect of CYP2D1 inhibition on the behavioural effects of d-amphetamine.

In rats, amphetamine (AMP) conversion to 4-OH-AMP is metabolized by CYP2D1, the rat equivalent of the human enzyme CYP2D6. To determine the impact of impaired AMP metabolism on its behavioural effects, AMP-induced hyperactivity, AMP discrimination and AMP self-administration were examined in male Wistar rats with or without pretreatment with the CYP2D1 inhibitors quinine and budipine. In vivo, quinine (20 mg/kg) and budipine (10 mg/kg) increased the plasma area under the curve of AMP 4-fold and 3.6-fold respectively, and decreased the plasma levels of 4-OH-AMP, 3-fold and 8.6-fold, confirming that the doses used suppressed CYP2D1 activity. Both inhibitors prolonged AMP-induced hyperactivity (0.3 mg/kg) and prolonged the duration of AMP-appropriate responding for periods of up to 90 min post-AMP administration in a drug discrimination procedure. In rats given a preload dose of AMP (0.8 mg/kg) 3 h prior to the self-administration test session, CYP2D1 inhibition resulted in fewer AMP infusions being taken compared with rats receiving the AMP preload dose alone. These studies indicate that AMP is responsible for the behavioural effects seen in rats and that a rat phenocopy model of the human CYP2D6 deficiency state can be produced by CYP2D1 inhibitors.

Alcohol Oxidoreductases↗
PubMed1996

Disruption of re-replication control by overexpression of human ORC1 in fission yeast.

Initiation of DNA replication in Saccharomyces cerevisiae requires the binding of the origin recognition complex (ORC) to autonomously replicating sequences. HsORC1, a recently identified human protein related to yeast Orc1p and Cdc6p/Cdc18p, may be a component of the replication initiation complex in human cells. We have independently isolated the gene for HsORC1 and begun to address its function in eukaryotic DNA replication and its relationship to Cdc18p. Although HsORC1 failed to rescue the temperature-sensitive S. pombe cdc18-K46 strain, overexpression in a wild-type strain led to continuous DNA synthesis in the absence of mitosis. Deletion mutagenesis identified a short N-terminal region of HsORC1 that contains potential phosphorylation sites for cyclin-dependent kinase (CDK) as being sufficient to induce re-replication. In addition, we found that HsORC1 is an efficient substrate for CDKs in vitro. We propose that perturbation of the re-replication control by overexpression of HsORC1 is due to a titration of components involved in inactivating Cdc18p upon initiation of replication.

Cell Compartmentation↗
PubMed1996

Phospholipase C beta 4 is involved in modulating the visual response in mice.

Expression of G protein-regulated phospholipase C (PLC) beta 4 in the retina, lateral geniculate nucleus, and superior colliculus implies that PLC beta 4 may play a role in the mammalian visual process. A mouse line that lacks PLC beta 4 was generated and the physiological significance of PLC beta 4 in murine visual function was investigated. Behavioral tests using a shuttle box demonstrated that the mice lacking PLC beta 4 were impaired in their visual processing abilities, whereas they showed no deficit in their auditory abilities. In addition, the PLC beta 4-null mice showed 4-fold reduction in the maximal amplitude of the rod a- and b-wave components of their electroretinograms relative to their littermate controls. However, recording from single rod photoreceptors did not reveal any significant differences between the PLC beta 4-null and wild-type littermates, nor were there any apparent differences in retinas examined with light microscopy. While the behavioral and electroretinographic results indicate that PLC beta 4 plays a significant role in mammalian visual signal processing, isolated rod recording shows little or no apparent deficit, suggesting that the effect of PLC beta 4 deficiency on the rod signaling pathway occurs at some stage after the initial phototransduction cascade and may require cell-cell interactions between rods and other retinal cells.

Animals↗
PubMed1996

Brain drug delivery of small molecules using immunoliposomes.

Immunoliposomes (antibody-directed liposomes) were used in the present study for delivery of the antineoplastic agent daunomycin to the rat brain. A coupling procedure was introduced, which allows conjugation of a thiolated antibody to maleimide-grafted 85-nm liposomes sterically stabilized with PEG. Antibody was thereby coupled to the terminal end of a PEG-conjugated linker lipid. No brain uptake of PEG-conjugated liposomes carrying [3H]daunomycin was observed. However, brain targeting of immunoliposomes carrying [3H]daunomycin was mediated by the OX26 monoclonal antibody to the rat transferrin receptor, which is selectively enriched at the brain microvascular endothelium that comprises the blood-brain barrier in vivo. Coupling of 30 OX26 antibodies per liposome resulted in optimal brain delivery. Saturation of delivery was observed at higher antibody densities. Determination of brain levels of immunoliposomes over 24 h revealed that immunoliposomes accumulate in brain tissue. Brain targeting of immunoliposomes was not observed in immunoliposomes conjugated with a mouse IgG2a isotype control. In addition, coinjection of free OX26 saturated plasma clearance of immunoliposomes. Since a single liposome may carry > or = 10,000 drug molecules, the use of PEG-conjugated immunoliposomes increases the drug carrying capacity of the monoclonal antibody by up to 4 logarithmic orders in magnitude. In summary, specific OX26-mediated targeting of daunomycin to the rat brain was achieved by the use of an immunoliposome-based drug delivery system.

Animals↗
PubMed1996

Ethanol cytotoxicity to a transfected HepG2 cell line expressing human cytochrome P4502E1.

The effect of ethanol on the viability of a HepG2 cell model which was developed to constitutively express human CYP2E1 was studied in an attempt to establish a linkage between CYP2E1, reactive oxygen intermediates, and ethanol toxicity. Assays of toxicity included leakage of lactate dehydrogenase, trypan blue uptake, morphology, and formazan production. Ethanol was toxic to HepG2 E9 cells, which express CYP2E1, but not to HepG2 MV5 cells, which do not express CYP2E1. The ethanol toxicity was dependent on the concentration of ethanol, starting with 10 m ethanol, and on the time of incubation with ethanol. Phorbol 12-myristate 13-acetate, which increases the expression of CYP2E1 in this model, increased the toxicity by ethanol. Ethanol toxicity was prevented by 4-methylpyrazole and by diallyl sulfide, inhibitors of CYP2E1. The ethanol toxicity was also prevented by radical trapping agents such as N-acetylcysteine and N-t-butyl-alpha-phenylnitrone, antioxidative agents such as catalase, superoxide dismutase, thiourea, and uric acid, and inhibitors of lipid peroxidation, such as vitamin E phosphate, Trolox, and diphenylphenylenediamine. Besides ethanol, other substrates such as Me2SO, CCl4, isoniazid, and N,N-dimethylnitrosamine were cytotoxic to cells expressing CYP2E1 but not to control cells. These results indicate that ethanol was toxic to HepG2 cells which express human CYP2E1 by a pathway sensitive to inhibitors of CYP2E1 and to a variety of antioxidative agents. This model appears to be useful in efforts to establish a CYP2E1-dependent ethanol hepatotoxicity system and to evaluate the role of oxidative stress and reactive radical species in the toxicity by ethanol.

Allyl Compounds↗
PubMed1996

Heterologous expression and characterization of recombinant Lactococcus lactis neutral endopeptidase (neprilysin).

A neutral endopeptidase (NEP) from Lactococcus lactis has recently been cloned and shown to contain high sequence homology with the human neutral endopeptidase, endopeptidase 24.11 (I. Mierau et al., J. Bacteriol. 175, 2087-2096, 1993). The gene for the neutral endopeptidase from L. lactis was cloned into the pQE expression vector, resulting in the fusion of a hexahistidine at the N-terminus. The recombinant enzyme was expressed to high levels in Escherichia coli (approximately 10 mg/liter of culture) and purified to homogeneity in a two-step procedure. A number of peptides were studied as substrates for the enzyme. The enzyme cleaves the following peptides at the Gly3-Phe4 bond: enkephalins, dynorphins A-6, A-8, A-9, A-10, A-13, and A-17, and alpha-neo-endorphin. In addition the enzyme hydrolyzes bradykinin, substance P, beta-endorphin, ACTH, and VIP. Although the cleavage patterns observed are similar to that seen with mammalian neutral endopeptidase, the lactococcal enzyme more efficiently cleaves larger peptide substrates. As observed with the mammalian neutral endopeptidase, the lactococcal enzyme exhibits higher kcat/K(m) values for the enkephalins than for their corresponding amides, indicating the functionality of an active-site arginine. Inactivation of the lactococcal endopeptidase by diethyl pyrocarbonate and protection afforded by the substrate dynorphin A-6 indicate the functionality of a positionally conserved active-site histidine. This was confirmed by demonstrating that conversion of this histidine, histidine 587, to glutamine generated inactive enzyme. Similarly, conversion of the putative zinc ligand glutamate 535 to glutamine led to inactive enzyme. These studies indicate a conservation of critical catalytic residues between the two enzymes and suggest that the lactococcal endopeptidase is a better model than thermolysin for the mammalian enzyme.

Amino Acid Sequence↗