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Biomedical subjects

F Wu

Publications and source records attributed to F Wu.

At least 73 records · Page 4Linked to original sources
PubMed2000

Cell-specific changes in expression of mRNAs encoding splice variants of aplysia cell adhesion molecule accompany long-term synaptic plasticity.

Aplysia neurons express several splice variants of apCAM, a member of the Ig superfamily of cell adhesion molecules. The major transmembrane isoform is endocytosed in sensory neurons (SNs) during the early phases of long-term facilitation (LTF) of SN synapses evoked by serotonin (5-HT) or in the motor neuron L7 during the early phases of long-term depression (LTD) of SN synapses evoked by Phe-Met-Arg-Phe-amide (FMRFa). We used single cell RT-PCR to evaluate whether expression of mRNAs encoding for different apCAM isoforms in SNs and L7 is regulated during LTF produced by 5-HT, and LTD produced by FMRFa. Single SNs and L7s express mRNAs encoding for all major isoforms, but the proportion of each isoform expressed differs for the two cells. SN expresses more mRNA encoding for GPI-linked isoforms, while L7 expresses more mRNA encoding for the major transmembrane isoform. The neuromodulators produced significant changes in the proportional levels of mRNAs encoding for specific apCAM isoforms during the first 4 h after treatments without affecting overall levels of apCAM mRNA. 5-HT evoked changes that exaggerated cell-specific differences in isoform expression. FMRFa evoked changes that reduced cell-specific differences in isoform expression. The effects of the neuromodulators on apCAM mRNA expression were not detected when cells were cultured alone or when SNs were cocultured with another motor cell that failed to induce synapse formation (L11). The results suggest that rapid cell-specific regulation of splice variant expression may contribute to different forms of long-term synaptic plasticity.

Alternative Splicing↗
PubMed2000

Capillary electrophoretic separation of the enantiomers of organophosphates with a phosphorus stereogenic center using the sodium salt of octakis(2,3-diacetyl-6-sulfo)-gamma-cyclodextrin as resolving agent.

The sodium salt of the single-isomer, chiral resolving agent, octakis(2,3-diacetyl-6-sulfo)-gamma-cyclodextrin (ODAS-gammaCD) has been used for the capillary electrophoretic separation of the enantiomers of alkylarylphosphates which carry a phosphorus-based stereogenic center. The effective mobilities and separation selectivities were measured at different ODAS-gammaCD and methanol concentrations to find the conditions under which the minor enantiomers could be adequately quantitated in samples obtained by chemical resolution of the racemic mixtures. This work extends the utility of ODAS-gammaCD to a hitherto unexplored field, the capillary electrophoretic separation of the enantiomers of organophosphorus compounds.

Cyclodextrins↗
PubMed2000

Interactions of the HIV-1 fusion peptide with large unilamellar vesicles and monolayers. A cryo-TEM and spectroscopic study.

We have examined the interaction of the human immunodeficiency virustype 1 fusion peptide (23 amino acid residues) and of a Trp-containing analog with vesicles composed of dioleoylphosphatidylcholine, dioleoylphosphatidylethanolamine and cholesterol (molar ratio, 1:1:1). Both the native and the Trp-substituted peptides bound the vesicles to the same extent and induced intervesicular lipid mixing with comparable efficiency. Infrared reflection-absorption spectroscopy data are compatible with the adoption by the peptide of a main beta-sheet structure in a cospread lipid/peptide monolayer. Cryo-transmission electron microscopy observations of peptide-treated vesicles reveal the existence of a peculiar morphology consisting of membrane tubular elongations protruding from single vesicles. Tryptophan fluorescence quenching by brominated phospholipids and by water-soluble acrylamide further indicated that the peptide penetrated into the acyl chain region closer to the interface rather than into the bilayer core. We conclude that the differential partition and shallow penetration of the fusion peptide into the outer monolayer of a surface-constrained bilayer may account for the detected morphological effects. Such single monolayer-restricted interaction and its structural consequences are compatible with specific predictions of current theories on viral fusion.

Acrylamide↗
PubMed2000

Corin, a transmembrane cardiac serine protease, acts as a pro-atrial natriuretic peptide-converting enzyme.

Atrial natriuretic peptide (ANP) is a cardiac hormone essential for the regulation of blood pressure. In cardiac myocytes, ANP is synthesized as a precursor, pro-ANP, that is converted to biologically active ANP by an unknown membrane-associated protease. Recently, we cloned a transmembrane serine protease, corin, that is highly expressed in the heart. In this study, we examine effects of corin on pro-ANP processing. Our results show that recombinant human corin converts pro-ANP to ANP and that the cleavage in pro-ANP by corin is highly sequence specific. Our findings suggest that corin is the long-sought pro-ANP-converting enzyme and that the corin-mediated pro-ANP activation may play a role in regulating blood pressure.

Animals↗
PubMed2000

Presynaptic morphological changes associated with long-term synaptic facilitation are triggered by actin polymerization at preexisting varicositis.

Morphological changes are thought to contribute to the expression of long-term synaptic plasticity, a cellular basis for learning and memory. The mechanisms mediating the initiation and maintenance of the morphological changes are poorly understood. We repeatedly imaged the axonal arbors of mechanosensory neurons of Aplysia as they formed new synaptic varicosities and axonal branches after applications of serotonin that cause long-term synaptic facilitation. New varicosities formed exclusively from preexisting varicosities, by splitting or branch outgrowth. These changes were prevented by cytochalasin D, which blocks actin polymerization and the turnover of actin filaments. The suppression of the morphological changes by cytochalasin D did not impair their expression when cytochalasin D was removed 24 hr after exposure to serotonin. These results imply that serotonin induces persistent effects at preexisting presynaptic varicosities, which enhance actin polymerization, and that this is essential for presynaptic morphological changes of long-term facilitation.

Actins↗
PubMed2000

Activation of the cytoplasmic c-Abl tyrosine kinase by reactive oxygen species.

The ubiquitously expressed c-Abl protein tyrosine kinase localizes to both the nucleus and cytoplasm. The nuclear form of c-Abl is activated in the cellular response to genotoxic stress. Here we show that cytoplasmic c-Abl is activated by oxidative stress. The results also demonstrate that mitochondrial cytochrome c is released in the cellular response to H(2)O(2) and that this effect is mediated by a c-Abl-dependent mechanism. In concert with these results, we show that H(2)O(2)-induced apoptosis is attenuated in c-Abl-deficient cells. These findings demonstrate that cytoplasmic c-Abl is involved in the apoptotic response of cells to oxidative stress.

Acetylcysteine↗
PubMed2000

Interaction between protein kinase C delta and the c-Abl tyrosine kinase in the cellular response to oxidative stress.

Protein kinase C (PKC) isoforms are phosphorylated on tyrosine in the response of cells to oxidative stress. The present studies demonstrate that treatment of cells with hydrogen peroxide (H(2)O(2)) induces binding of the PKCdelta isoform and the c-Abl protein-tyrosine kinase. The results show that c-Abl phosphorylates PKCdelta in the H(2)O(2) response. We also show that PKCdelta phosphorylates and activates c-Abl in vitro. In cells, induction of c-Abl activity by H(2)O(2) is attenuated by the PKCdelta inhibitor, rottlerin, and by overexpression of the regulatory domain of PKCdelta. These findings support a functional interaction between PKCdelta and c-Abl in the cellular response to oxidative stress.

Animals↗
PubMed2000

Bidentate ligands that contain pyrrole in place of pyridine.

A series of ligands are prepared that are analogues of benzo-fused derivatives of 2,2'-bipyridine (bpy), in which pyrrole has been substituted for a pyridine ring. These ligands include 2-(2'-pyridyl)-indole, 2-(2'-pyrrolyl)-quinoline, and pyrrolo[3,2-h]quinoline. A novel reductive cyclization approach to the latter species is presented. All these ligands react with [Ru(bpy-d8)2Cl2], undergoing cyclometalation with concurrent deprotonation, to form complexes of the type [Ru(L)(bpy-d8)2]+ where L binds as a monoanionic ligand. The complexes are readily characterized by their 1H NMR spectra. Changes in the redox potentials and the electronic absorption spectra of both the ligands and the complexes are interpreted in terms of charge delocalization on the ligand.

Journal Article↗
PubMed2000

Distribution of (76)Br-labeled antisense oligonucleotides of different length determined ex vivo in rats.

Oligonucleotides may hybridize with high selectivity to an RNA sequence and can be used for the monitoring of gene expression or for its inhibition in experimental or therapeutic purposes. As part of the development of positron emission tomography (PET) methods, different lengths (30, 20, 12 and 6 mer) of antisense phosphorothioate oligonucleotides complementary to rat chromogranin A were labeled with [(76)Br] using a prosthetic group. The (76)Br-oligonucleotides were injected into rat's tail vein (1-2 MBq/rat), and the radioactivity distribution was analyzed after 20 h using whole body autoradiography or by measurement of organ radioactivity concentration. The whole body autoradiography showed different distribution depending on the oligonucleotide length. The organs with highest uptake changed from kidney cortex (with 6 or 12 mer), kidney cortex and liver (with 20 mer), to liver and spleen (with 30 mer). With 20 or 30 mer sequences, uptake could be observed in the adrenals. Kidneys and livers from rats receiving 20 mer or 30 mer (76)Br-oligonucleotides were analyzed with respect to subcellular distribution and DNA/RNA/protein fraction. 30%-45% of the radioactivity was found in the nuclear fraction. More than 80% of the radioactivity was recovered in the high molecular weight fraction (as proteins or oligonucleotides longer than 10 mer) using size exclusion (NAP 5) gelfiltration or cetylpyridinium bromide (CPB) precipitation. This work indicates the potential to perform kinetic whole body studies of (76)Br-oligonucleotides using PET.

Animals↗
PubMed2000

Dynamical simulation of LACBED patterns in cross-sectioned heterostructures

The Large Angle Convergent Beam Electron Diffraction (LACBED) technique has been applied to determination of the tetragonal mismatch in coherent Si/Si1-xGex/Si heterostructures. Two-dimensional (2D) dynamical simulation of the LACBED patterns has been performed and compared with the corresponding experimental ones. A good agreement is found in the whole simulated area, particularly as regards the splitting of the Bragg contours, due to the strain field present in the TEM cross-sections.

Journal Article↗
PubMed2000

Control of arterial blood pressure and renal sodium excretion by nitric oxide synthase in the renal medulla.

Work in our laboratory has focused on the role of nitric oxide synthase (NOS) in the regulation of renal medullary function. Biochemical studies demonstrated that the renal medulla is enriched in immunoreactive NOS protein and NOS enzymatic activity when compared with the renal cortex. Further experiments showed large amounts of NOS activity in the inner medullary collecting ducts, while moderate NOS activity was found in glomeruli and vasa recta and minimal NOS activity was detected in other nephron segments examined. In subsequent functional studies, selective renal medullary infusion of NOS stimulators (bradykinin or acetylcholine) or inhibitors (L-NAME) preferentially altered medullary blood flow. The alterations in medullary flow were associated with parallel changes in sodium and water excretion. Similar to the effects observed in anaesthetized rats, chronic infusion of L-NAME directly into the renal medullary interstitial space of conscious, uninephrectomized SD rats selectively decreased renal medullary blood flow throughout a 5-day L-NAME infusion period. The decrease in medullary blood flow was associated with retention of sodium and the development of hypertension, and the effects were reversible. In contrast to the effects of chronic NOS inhibition, renal medullary infusion of NOS substrate L-arginine prevented the development of sodium-sensitive hypertension in the Dahl salt-sensitive rat placed on a high sodium diet. The data reviewed in this paper indicate that NOS isoforms expressed in the renal medulla have a potent influence on renal medullary tubular and vascular function with consequential effects on fluid and electrolyte homeostasis and arterial blood pressure.

Animals↗
PubMed2000

Tubal sterilization and use of an IUD and risk of cervical cancer.

The relationship of tubal sterilization and use of an intrauterine device (IUD) to the risk of invasive squamous cell cervical cancer was evaluated in a case-control study carried out in Shandong, China, from 1989 to 1991. Patients (cases) were 272 women aged 30-77 years with newly diagnosed invasive squamous cell cervical cancer in Shandong Province Tumor Hospital. Controls were 893 randomly selected screened women matched to the cases by age (within 2 years) and county. A decrease in risk was observed in uses of an IUD, especially in long-term users under age 33. A nonsignificant decrease in risk in women under age 33 who had had a tubal ligation was also observed, especially in the first 10 years since the procedure. Tubal ligation and IUD insertion and removal provide opportunities to screen women for cervical cancer and reduce the risk of invasive disease.

Adult↗
PubMed2000

An integrated risk model of a drinking-water-borne cryptosporidiosis outbreak.

A dynamic risk model is developed to track the occurrence and evolution of a drinking-water-borne cryptosporidiosis outbreak. The model characterizes and integrates the various environmental, medical, institutional, and behavioral factors that determine outbreak development and outcome. These include contaminant delivery and detection, water treatment efficiency, the timing of interventions, and the choices that people make when confronted with a known or suspected risk. The model is used to evaluate the efficacy of alternative strategies for improving risk management during an outbreak, and to identify priorities for improvements in the public health system. Modeling results indicate that the greatest opportunity for curtailing a large outbreak is realized by minimizing delays in identifying and correcting a drinking-water problem. If these delays cannot be reduced, then the effectiveness of risk communication in preemptively reaching and persuading target populations to avoid exposure becomes important.

Choice Behavior↗
PubMed2000

Characterization of L-arginine transporters in rat renal inner medullary collecting duct.

Previous work from our laboratory has demonstrated that the inner medullary collecting duct (IMCD) expresses a large amount of nitric oxide synthase (NOS) activity. The present study was designed to characterize the transport of NOS substrate, L-arginine, in a suspension of bulk-isolated IMCD cells from the Sprague-Dawley rat kidney. Biochemical transport studies demonstrated an L-arginine transport system in IMCD cells that was saturable and Na(+) independent (n = 6). L-Arginine uptake by IMCD cells was inhibited by the cationic amino acids L-lysine, L-homoarginine, and L-ornithine (10 mmol/l each) and unaffected by the neutral amino acids L-leucine, L-serine, and L-glutamine. Both L-ornithine (n = 6) and L-lysine (n = 6) inhibited NOS enzymatic activity in a dose-dependent manner in IMCD cells, supporting the important role of L-arginine transport for NO production by this tubular segment. Furthermore, RT-PCR of microdissected IMCD confirmed the presence of cationic amino acid transporter CAT1 mRNA, whereas CAT2A, CAT2B, and CAT3 were not detected. These results indicate that L-arginine uptake by IMCD cells occurs via system y(+), is encoded by CAT1, and may participate in the regulation of NO production in this renal segment.

Amino Acid Transport Systems↗
PubMed2000

Nitric oxide synthase activity and isoforms in rat renal vasculature.

Experiments were performed to quantify nitric oxide synthase (NOS) activity and identify the NOS isoforms present in the Sprague-Dawley rat renal vasculature. NOS enzymatic activity was measured by adding [(3)H]arginine to microdissected renal blood vessels and quantifying the conversion to [(3)H]citrulline by reverse-phase high-performance liquid chromatography. Total NOS activity was greatest in microdissected vasa recta (123+/-41 pmol. mg(-1). h(-1), n=5) and significantly less in glomeruli (46+/-9 pmol. mg(-1). h(-1), n=6) and afferent arterioles (42+/-10 pmol. mg(-1). h(-1), n=6) and averaged <5 pmol. mg(-1). h(-1) in arcuate (n=8) and interlobular (n=9) arteries. Addition of 1.0 mmol/L EDTA to the reaction decreased NOS activity to <5 pmol. mg(-1). h(-1) in afferent arterioles, glomeruli, and vasa recta (n=5 each), indicating that the NOS enzymatic activity in these segments is primarily a result of constitutive NOS. Both neuronal and endothelial NOS mRNA were identified in each vascular segment by reverse transcription-polymerase chain reaction, but inducible NOS mRNA was detected only in microdissected arcuate arteries. The present experiments indicate that the vasa recta, glomeruli, and afferent arterioles contain large amounts of calcium-dependent NOS enzymatic activity and that neuronal NOS and endothelial NOS mRNA are present in these segments.

Animals↗
PubMed2000

[Different flow augmentations in coronary, carotid and renal arteries with ECP in experimental canines].

Different flow augmentations were found in canine's coronary, carotid and renal arteries with ECP because of their different anatomical locations in aorta. We used the Electromagnetic Blood Flowmeters to measure the blood flow pre-ECP and during ECP. The results showed ECP increased the diastolic flow of coronary, carotid and renal arteries by 24.48% +/- 1.29%, 27.18% +/- 0.94% and 20.68% +/- 0.80% respectively, decreased their systolic flow by 4.77% +/- 0.48%, 6.45% +/- 0.99% and 9.60% +/- 0.70%, but augmented their mean flow by 18.24% +/- 2.01%, 19.38% +/- 1.47% and 8.42% +/- 0.94% respectively in experimental dogs. These data demonstrate that ECP can greatly increase the mean flow of coronary and carotid arteries but at the same time patently decrease the systolic flow of renal artery.

Animals↗