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Biomedical subjects

W Feng

Publications and source records attributed to W Feng.

At least 37 records · Page 2Linked to original sources

Arthropod- and host-specific Borrelia burgdorferi bbk32 expression and the inhibition of spirochete transmission.

Antisera to BBK32 (a Borrelia burgdorferi fibronectin-binding protein) and BBK50, two Ags synthesized during infection, protect mice from experimental syringe-borne Lyme borreliosis. Therefore, B. burgdorferi bbk32 and bbk50 expression within Ixodes scapularis ticks and the murine host, and the effect of BBK32 and BBK50 antisera on spirochetes throughout the vector-host life cycle were investigated. bbk32 and bbk50 mRNA and protein were first detected within engorged ticks, demonstrating regulated expression within the vector. Then bbk32 expression increased in mice at the cutaneous site of inoculation. During disseminated murine infection, bbk32 and bbk50 were expressed in several murine tissues, and mRNA levels were greatest in the heart and spleen at 30 days. BBK32 antisera protected mice from tick-borne B. burgdorferi infection and spirochete numbers were reduced by 90% within nymphs that engorged on immunized mice. Moreover, 75% of these ticks did not retain spirochetes upon molting, and subsequent B. burgdorferi transmission by adult ticks was impaired. Larval acquisition of B. burgdorferi by I. scapularis was also inhibited by BBK32 antisera. These data demonstrate that bbk32 and bbk50 are expressed during tick engorgement and that BBK32 antisera can interfere with spirochete transmission at various stages of the vector-host life cycle. These studies provide insight into mechanisms of immunity to Lyme borreliosis and other vector-borne diseases.

Animals↗

Effects of CTx and 8-bromo-cAMP on LPS-induced gene expression of cytokines in murine peritoneal macrophages.

LPS, an endotoxin isolated from gram-negative bacterial, has been shown to be a potent cytokine initiator in murine peritoneal macrophages. CAMP-dependent pathway is generally considered to play a suppressive role in immune response. This study investigated the effect of cAMP on LPS-induced gene expression of cytokines in murine macrophages. Our data clearly demonstrated that in LPS-treated macrophages, cAMP elevator (CTx and 8-bromo-cAMP) could increase IL-1Ra and IL-10 gene expression, while mRNAs of IL-1alpha, IL-12, IL-6, and MIF were decreased and other cytokines like IL-1beta, and IFN-gamma did not give a definite tendency. This is the first report that CTx and 8-bromo-cAMP positively regulate IL-1Ra gene expression in LPS-stimulated macrophages. Our data also suggest that a cAMP-dependent pathway may play a regulatory role in Toll-receptor system.

8-Bromo Cyclic Adenosine Monophosphate↗

Photochemistry of hydrolytic iron (III) species and photoinduced degradation of organic compounds. A minireview.

On the basis of the research literature of the last fifty years, we have reviewed the photochemical properties of different hydrolytic Fe(III) species and the photodegradation of organic compounds in aqueous solutions initiated by them (mainly including low-molecular-weight Fe(III)-OH complexes, Fe(III) oxides (Fe2O3) and Fe(III) hydroxides (FeOOH)). Furthermore, the feasibility and prospects of the novel photochemical methods of using Fe(III) salts and oxides in wastewater treatments are being discussed in this paper.

Hydrolysis↗

Degradation mechanism of azo dye C. I. reactive red 2 by iron powder reduction and photooxidation in aqueous solutions.

We have made a comparison of the UV-VIS spectra of three azo dyes, C. I. reactive red 2, orange II and C. I. reactive black 8, in aqueous solutions during treatment with iron powder reduction and photooxidation. From this, we propose their mechanisms for reduction photooxidation. GC/MS analyses of the degradation products of the dye C. I. reactive red 2 demonstrated some important steps producing hydrogenated azo structure, substituted benzene and substituted naphthalene.

Azo Compounds↗

Functional role of hyperreactive sulfhydryl moieties within the ryanodine receptor complex.

Several laboratories using chemically heterogeneous sulfhydryl modifying agents have shown that sarcoplasmic reticulum (SR) Ca2+ channels known as ryanodine receptors (RyRs) are especially sensitive to modification of functionally important cysteine residues. The functional consequence of sulfhydryl modification of RyRs can include phases of activation and inhibition that are very much dependent on the concentration of the reagent used, the length of exposure, and the nature of the chemical reaction the reagent undertakes with sulfhydryl groups. Most challenging is understanding the relationship for how specific sulfhydryl moieties ascribe specific aspects of RyR function. Considering the structural complexity of the RyR complex with its associated proteins, this task is likely to be a formidable one. A small number of hyperreactive thiols have been shown to exist within the RyR complex. Their functional role does not appear to impact directly on channel gating. Rather hyperreactive cysteine (Cys) moieties may represent biochemical components of a redox sensor that conveys information about localized changes in redox potential produced by physiologic (e.g., glutathione, nitric oxide) and pathophysiologic (quinones, reactive oxygen species) channel modulators to the Ca2+ release process. The molecular and functional details of such a redox sensor remains to be elucidated.

Animals↗

Effect of endurance training on lipid metabolism in women: a potential role for PPARalpha in the metabolic response to training.

Endurance training increases fatty acid oxidation (FAO) and skeletal muscle oxidative capacity. However, the source of the additional fat and the mechanisms for increasing FAO capacity in muscle are not clear. We measured whole body and regional lipolytic activity and whole body and plasma FAO in six lean women during 90 min of bicycling exercise (50% pretraining peak O(2) consumption) before and after 12 wk of endurance training. We also assessed skeletal muscle content of peroxisome proliferator-activated receptor-alpha (PPARalpha) and its target proteins that regulate FAO [medium-chain and very long chain acyl-CoA dehydrogenase (MCAD and VLCAD)]. Despite a 25% increase in whole body FAO during exercise after training (P < 0.05), training did not alter regional adipose tissue lipolysis (abdominal: 0.56 +/- 0.26 and 0.57 +/- 0.10 micromol x 100 g(-1) x min(-1); femoral: 0.13 +/- 0.07 and 0.09 +/- 0.02 micromol x 100 g(-1) x min(-1)), whole body palmitate rate of appearance in plasma (168 +/- 18 and 150 +/- 25 micromol/min), and plasma FAO (554 +/- 61 and 601 +/- 45 micromol/min). However, training doubled the levels of muscle PPARalpha, MCAD, and VLCAD. We conclude that training increases the use of nonplasma fatty acids and may enhance skeletal muscle oxidative capacity by PPARalpha regulation of gene expression.

Acyl-CoA Dehydrogenase↗

[Advances in diagnosis and treatment of ectopic pregnancy during the past ten years].

OBJECTIVE: To investigate the changes of diagnosis and treatment of ectopic pregnancy (EP) during the past ten years. METHODS: Medical records of 1,970 cases with EP seen in our hospital from Jan. 1987 to Dec. 1996 were collected and analyzed retrospectively. RESULTS: (1) The incidence of EP increases from 1.0 per 42.4 labours during 1987-1988 to 1.0 per 6.6 labours in 1995-1996. (2) Diagnosis was confirmed more by television laparoscopy, transvaginal ultrasonography and sensitive human chorionic gonadotropin-beta subunit assay rather than loparotomy and with a shorter period, (6.5 +/- 0.7) days in 1987-1988 versus (3.7 +/- 0.2) days in 1995-1996. (3) The choice of treatment was more conservative or through laparoscopic surgery than abdominal salpingectomy. (4) The success rate of conservative surgery expectant management reached 100.0% and medical treatment with a variety of choices around 80.0%-85.%. CONCLUSIONS: Although the incidence of EP increased, with the improvement of diagnostic approaches, patients were detected at an earlier stage and possible to be treated more conservatively.

Female↗

Modification of ryanodine receptor/Ca2+ release channel with dinitrofluorobenzene.

Modification of the ryanodine receptor (RyR)/Ca(2+) release channel with 2,4-dinitrofluorobenzene (DNFB) indicated that two classes of amino group interact with the reagent, as can be distinguished on the basis of their reactivity/accessibility and the effects on ryanodine binding and single channel activities. One group interacted very rapidly (t(1/2)<30 s) at 25 degrees C with low concentrations of DNFB [C(50) (concentration of DNFB required for 50% inhibition or stimulation of ryanodine binding)=5 microM], and at pH values of 6.2 and higher. This interaction resulted in the marked stimulation of ryanodine binding and the complete inhibition of a single Ca(2+) release channel incorporated into planar lipid bilayer. The second group is accessible at higher temperatures (37 degrees C); at pH values higher than 7.4 it reacted slowly (t(1/2)=20 min) with high concentrations of DNFB (C(50)=70 microM). This interaction led to the inhibition of ryanodine binding and single channel activity. Modification of RyR with DNFB under the stimulatory conditions resulted in 3.6-fold and 6-fold increases in ryanodine-binding and Ca(2+)-binding affinities respectively. Modification with DNFB under the inhibitory conditions resulted in a decrease in the total ryanodine-binding sites. The exposure of the RyR single channel to DNFB under both inhibitory and stimulatory conditions led to the complete closure of the channel. However, when modified under the stimulatory conditions, but not under the inhibitory ones, the DNFB-modified closed channel could be re-activated by sub-micromolar concentrations of ryanodine, in the presence of nanomolar concentrations of Ca(2+). The DNFB-modified ryanodine-activated RyR channel showed fast transitions between open, closed and several sub-conductance states, and was completely closed by Ruthenium Red. ATP re-activated the DNFB-modified closed channel or, if present during modification, prevented the inhibition of RyR channel activity by DNFB. Neither the stimulation nor the inhibition of ryanodine binding by modification with DNFB was affected by the presence of ATP. By using the photoreactive ATP analogue 3'-O-(4-benzoyl)benzoyl-[alpha-(32)P]ATP we found that DNFB modification had no effect on the ATP-binding site of RyR. The results are discussed with regard to the involvement of amino group residues in channel gating, ryanodine association/dissociation and occlusion, and the relationship between the open/closed state of the RyR and its capacity to bind ryanodine.

Adenosine Triphosphate↗

Affinity chromatography of human estrogen receptor-alpha expressed in Saccharomyces cerevisiae. Combination of heparin- and 17beta-estradiol-affinity chromatography.

Estrogen receptor-alpha is a member of the nuclear hormone receptor superfamily and is considered as a very important regulatory protein. Human estrogen receptor-alpha has been cloned into Saccharomyces cerevisiae as a fusion to ubiquitin and expression is controlled by a metallothionin promotor. Pilot scale quantities of receptor have been produced by a yeast strain transformed with expression plasmid YEpE13 [Graumann et al., J. Steroid Biochem. Mol. Biol. 57 (1996) 293] in a 14 l stirred tank reactor. The yeast extract contained 2-4 pmol of receptor protein per mg total protein. A purification scheme has been developed using heparin-affinity chromatography combined with affinity chromatography with immobilized 17beta-estradiol 17-hemisuccinate. Heparin-affinity chromatography was very efficient to remove host cell protein. Accompanying proteins that stabilize unoccupied receptor have not been dissociated during elution. The receptor could be purified 5-10-fold in ligand-free state. In contrast to previous reports, we did not find a difference of the binding affinity of liganded and unliganded receptor for heparin immobilized onto Sepharose. The unoccupied receptor could be further purified 100-fold with ligand-affinity chromatography using 17beta-estradiol 17-hemisuccinate-bovine serum albumin-Sepharose. The receptor could be kept in its native state, although saturated with 17beta-estradiol. The purification sequence allows an efficient production of receptor. Further improvement of productivity can be only accomplished by increasing the expression level.

Base Sequence↗

Effects of dextran on tracheal mucociliary velocity in dogs in vivo.

We have previously shown that dextran (molecular weight 4kDa) is a potential mucolytic agent, reducing the viscoelasticity and spinnability of cystic fibrosis (CF) sputum and improving its mucociliary clearability during in vitro testing. We wished to see whether low molecular weight (LMW) dextran had similar effects on mucus rheology when administered by aerosol to living dogs, and whether the administration of dextran increased the rate of mucociliary clearance. Healthy mongrel dogs were anesthetized with pentobarbital and intubated. After a 30-min Ringer aerosol delivery during spontaneous breathing, tracheal mucociliary velocity (TMV by charcoal marker particle transport) was measured under bronchoscopic control, and mucus for viscoelasticity analysis (magnetic rheometer) was collected by the endotracheal tube method. Then LMW dextran in Ringer vehicle was delivered by aerosol via the endotracheal tube, followed by the same procedures. We performed eight experiments in eight dogs, involving 30 min administrations of dextran aerosol; all dogs received inhalations of 20 mg/ml, 65 mg/ml, and 200 mg/ml dextran. Compared with Ringer control, TMV increased to 145% of control (P=0.0417) at 65 mg/ml dextran. Mucus viscoelasticity (G*) significantly decreased to 19% of control (P=0.0426) at 65 mg/ml. This in vivo study supports our previous in vitro testing, that LMW dextran decreases the mucus viscoelasticity and increases the rate of mucociliary clearance. We estimate the dosage received by aerosol at 65 mg/ml to be within the effective concentration range studied in vitro, i.e. 10-15 mg/ml final concentration. The results are consistent with the proposed mechanism that the saccharide moieties in LMW dextran compete for hydrogen bonding sites with other mucous glycoproteins. These new hydrogen bonds are structurally and rheologically ineffective, thus reducing the overall cross-link density, and making the mucus more easily cleared by ciliary and cough mechanisms.

Administration, Inhalation↗

Comparison of the chromium distribution in organs and subcellular fractions of normal and diabetic rats by using enriched stable isotope Cr-50 tracer technique.

The enriched stable isotope 50Cr(III) tracer technique combined with neutron activation analysis was used to examine the intracellular distribution of Cr(III) in the liver, pancreas, testes, and kidney homogenates of both normal and diabetic rats. Our new results showed that the nucleic fraction has the highest Cr concentration in the liver cell of both normal and diabetic rats. The diabetic rats retain more Cr in the mitochondrial and lysosomal fractions of liver homogenate than the normal. This is likely an indication of chromium participating in the glucose or lipid metabolism to compensate the low level of insulin in the body of diabetic rats. The concentrations of Cr in the subcellular fractions of pancreas, testes, and kidney in the normal rats are higher than those in the diabetic rats, which favor the hypothesis that Cr(III) plays its biological function via interaction with the insulin-sensitive tissues or enhancement of the sensitivity of the insulin receptor.

Animals↗

Borrelia burgdorferi erpT expression in the arthropod vector and murine host.

The expression of a Borrelia burgdorferi gene, erpT, was investigated throughout the spirochaete life cycle in the arthropod vector and the murine host. Three phage clones from a B. burgdorferi DNA expression library synthesized a 30 kDa antigen that was recognized by antibodies in the sera of B. burgdorferi-infected mice but not mice hyperimmunized with B. burgdorferi lysates. Differential antibody binding suggested that this protein was preferentially expressed in vivo. This antigen was designated ErpT, based upon 99.6% homology with the BBF01 sequence in the B. burgdorferi genome. ErpT was not detected on spirochaetes cultured in BSK II medium by indirect immunofluorescence or in B. burgdorferi lysates by immunoblotting, implying that ErpT is not readily produced in vitro. erpT mRNA was not discernible by Northern blot but was identified by RNA polymerase chain reaction in vitro, indicating that erpT is expressed at low levels by cultured spirochaetes. erpT expression was then investigated in the vector and mice because B. burgdorferi do not normally reside in culture medium. RNA polymerase chain reaction and immunofluorescence studies demonstrated that erpT was expressed by a small minority of B. burgdorferi (11/500, 2.2%) within unfed ticks and then repressed during engorgement. erpT mRNA or ErpT antibodies were first detected in B. burgdorferi-infected mice at 4 weeks, suggesting that erpT was not expressed in the early stages of murine infection. Then, during persistent infection, RNA polymerase chain reaction showed that erpT was expressed by B. burgdorferi within the joints, heart and spleen, but not by spirochaetes in the skin. Immunization of mice with ErpT was antigenic but was not protective. These studies demonstrate that B. burgdorferi erpT is differentially expressed throughout the B. burgdorferi life cycle, in both the vector and the mammalian host, and is primarily expressed in extracutaneous sites during murine infection.

Animals↗

Antagonistic effects of NES and NLS motifs determine S. cerevisiae Rna1p subcellular distribution.

Nucleus/cytosol exchange requires a GTPase, Ran. In yeast Rna1p is the GTPase activating protein for Ran (RanGAP) and Prp20p is the Ran GDP/GTP exchange factor (GEF). RanGAP is primarily cytosolic and GEF is nuclear. Their subcellular distributions led to the prediction that Ran-GTP hydrolysis takes place solely in the cytosol and GDP/GTP exchange solely in the nucleus. Current models propose that the Ran-GTP/Ran-GDP gradient across the nuclear membrane determines the direction of exchange. We provide three lines of evidence that Rna1p enters and leaves the nuclear interior. (1) Rna1p possesses leucine-rich nuclear export sequences (NES) that are able to relocate a passenger karyophilic protein to the cytosol; alterations of consensus residues re-establish nuclear location. (2) Rna1p possesses other sequences that function as a novel nuclear localization sequence able to deliver a passenger cytosolic protein to the nucleus. (3) Endogenous Rna1p location is dependent upon Xpo1p/Crm1p, the yeast exportin for leucine-rich NES-containing proteins. The data support the hypothesis that Rna1p exists on both sides of the nuclear membrane, perhaps regulating the Ran-GTP/Ran-GDP gradient, participating in a complete RanGTPase nuclear cycle or serving a novel function.

Amino Acid Sequence↗

Site-selective modification of hyperreactive cysteines of ryanodine receptor complex by quinones.

Quinones undergo redox cycling and/or arylation reactions with key biomolecules involved with cellular Ca2+ regulation. The present study utilizes nanomolar quantities of the fluorogenic maleimide 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM) to measure the reactivity of hyperreactive sulfhydryl moieties on sarcoplasmic reticulum (SR) membranes in the presence and absence of quinones by analyzing the kinetics of forming CPM-thioether adducts and localization of fluorescence by SDS-polyacrylamide gel electrophoresis. Doxorubicin, 1,4-naphthoquinone (NQ), and 1, 4-benzoquinone (BQ) are found to selectively and dose-dependently interact with a class of hyperreactive sulfhydryl groups localized on ryanodine-sensitive Ca2+ channels [ryanodine receptor (RyR)], and its associated protein, triadin, of skeletal type channels. NQ and BQ are the most potent compounds tested for reducing the rate of CPM labeling of hyperreactive SR thiols (IC50 = 0.3 and 1.8 microM, respectively) localized on RyR and associated protein. The reduced forms of quinone, tert-butylhydroquinone, and 5-imino-daunorubicin do not alter significantly the pattern or kinetics of CPM labeling up to 100 microM, demonstrating that the quinone group is essential for modulating the state of hyperreactive SR thiols. Nanomolar NQ is shown to enhance the association of [3H]ryanodine for its high-affinity binding site and directly enhance channel-open probability in bilayer lipid membrane in a reversible manner. By contrast, micromolar NQ produces a time-dependent biphasic action on channel function, leading to irreversible channel inactivation. These results provide evidence that nanomolar quinone selectively and reversibly alters the redox state of hyperreactive sulfhydryls localized in the RyR/Ca2+ channel complex, resulting in enhanced channel activation. The Ca2+-dependent cytotoxicities observed with reactive quinones formed at the microsomal surface by oxidative metabolism may be related to their ability to selectively modify hyperreactive thiols regulating normal functioning of microsomal Ca2+ release channels.

Animals↗

[An experimental study of anti-angiogenesis with a cartilage-derived inhibitor].

OBJECTIVE: To evaluate the efficacy of a cartilage-derived inhibitor for the treatment of corneal neovascularization in vivo. METHODS: A cartilage-derived inhibitor (CDI) from bovine scapula was purified to homogeneity. By rabbit corneal neovascularization (CNV) model, the effect of inhibition of CDI under various conditions was determined in corneal micropocket analysis. RESULTS: The purified CDI could inhibit strongly the growing speed and area of rabbit CNV compared to control (P < 0.001). CONCLUSION: It is demonstrated that CDI is a potent dose-dependent inhibitor of angiogenesis.

Angiogenesis Inhibitors↗

[The selection of tympanoplastic type in treating chronic suppurative otitis media].

OBJECTIVE: To investigate the effect of different type of tympanoplasty on chronic suppurative otitis media (CSOM). METHOD: Myringoplasty, focuses clearance and myringoplasty, focuses clearance and ossiculoplasty and myringoplasty were performed in 189 cases of CSOM. RESULT: The cure rate of perforation of tympanic membrane with myringoplasty was above 99% accompanied by the varying improvement in acoustic sensibility. CONCLUSION: Different types of tympanoplasty should be used according to the CSOM. The slighter the pathological changes of CSOM, the more the hearing improve. The better effects was in tympanoplasty of simple type. The tympanic exploration is necessary to be included in tympanoplasty.

Adolescent↗

The Kaiser Permanente/USC Patient Consultation Study: change in use and cost of health care services.

The impacts of three alternative models of pharmacist consultation on the use and cost of health care services were studied. Two studies were conducted concurrently in an HMO over two years. In one, 6000 patients were randomly assigned to one of three consultation models; in the other, the three models were implemented in six geographic regions of California (4600 patients). The models were (1) consultation about new or changed prescriptions as mandated by state law (state model), (2) consultation focused on selected high-risk ambulatory care patients (Kaiser Permanente [KP] model), and (3) a control model. The patients were surveyed three times about their health status and satisfaction, and computerized data on health care use and cost were collected. The effect of the consultation models on the use and cost of health care services was examined across five risk groups that were based on drug-use profiles. An additional 37,750 patients (10% of the patients residing in the areawide study sites) were included in a supplemental analysis of the use and cost of health care services. There was no indication in the random-assignment study that pharmacist consultations affected either drug costs or the cost of office visits. Similar results were found in the areawide study, with the exception that the KP model was associated with lower drug costs than the control model. In the 10% sample, the KP model appeared to be associated with lower office visit costs but higher drug costs. Both models were associated with a lower likelihood of a hospital admission and with lower total health care costs for some high-risk patients compared with the control model. Counseling patients about their medications may be unlikely to reduce medication costs or the cost of office visits but may reduce the likelihood of hospital admissions and the overall costs of health care services; a combination of counseling patients at high risk for drug-related problems and counseling all patients about any new or changed prescription should be considered.

Adult↗