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Biomedical subjects

J Fu

Publications and source records attributed to J Fu.

At least 217 records · Page 12Linked to original sources

TEMPOL, a superoxide dismutase mimic, ameliorates light-induced retinal degeneration.

The efficacy of 4-hydroxy-2,2,6,6-tetramethylpiperidine-l-oxyl (TEMPOL), a metal independent superoxide dismutase (SOD) mimic, in ameliorating light-induced retinal degeneration was investigated. Thirty-six Lewis albino rats were exposed to green fluorescent light (490-580 nm, 160-180 foot-candles) for 24 hr, after dark adaptation for 24 hr. The animals received six intraperitoneal (IP) injections of TEMPOL (100 mg/kg) or an equivalent volume of saline solution (vehicle-treated control groups) at 6 hr intervals starting 6 hr before the light exposure and ending 24 hr after light exposure. Another six rats were used as unexposed controls. The animals were killed at 6 hr, 6 days and 14 days after light exposure. Retinal damage was assessed by light and electron microscopy, measurements of outer nuclear layer (ONL) thickness and rhodopsin levels and counting of macrophages in the subretinal space. After light exposure, the TEMPOL-treated rats showed mild edema of the retinal pigment epithelium (RPE), less densified inner segments (IS) at 6 hr and better preserved photoreceptors at 6 days and 14 days compared with vehicle-treated control groups. Morphometrically the ONL was thicker in the TEMPOL-treated rats than in the vehicle-treated control at 6 days (p<0.01) and 14 days (p<0.05) but no significant difference occurred at 6 hr (p>0.05). Rhodopsin levels in the TEMPOL-treated rats were significantly higher at 6 days (p<0.05) but not at 6 hr (p>0.05) or 14 days (p>0.05). Our results demonstrated that TEMPOL ameliorated light-induced retinal degeneration in rats. These findings are consistent with the hypothesis that superoxide radicals may play a crucial role in mediating light-induced retinal degeneration.

Animals↗

Purification and characterization of HIV-1 reverse transcriptase having a 1:1 ratio of p66 and p51 subunits.

Wild-type and several mutant forms of recombinant human immunodeficiency virus type-1 reverse transcriptase were overexpressed as either the p66 or the p51 subunit in a protease-deficient strain of Escherichia coli. Immediately prior to cell lysis, p51 cell paste was mixed with cell paste containing the corresponding overexpressed p66 subunit in a ratio resulting in an excess of the smaller subunit with respect to the larger. During the subsequent chromatography steps stable heterodimer p66/p51 was purified to homogeneity. This protein was characterized by amino acid analysis, denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, analytical gel filtration HPLC, laser desorption mass spectroscopy, and isoelectric focusing. In addition, we were able to obtain crystals of the purified enzyme complexed with a quinazolinone class nonnucleoside inhibitor that diffracted to 3.2 A resolution. A potential application of this expression/purification methodology is the ability to alter specific amino acids residues, by site-directed-mutagenesis, of only one subunit of the RT-dimer.

Base Sequence↗

Intravitreal delivery of ganciclovir in rabbits by transscleral iontophoresis.

To avoid the side effects of systemic administration of ganciclovir (GCV) for the treatment of cytomegalovirus (CMV) retinitis, we studied transscleral iontophoresis of GCV into rabbit eyes. After a single application with 20% (w/w) aqueous solution of GCV at 1.0 mA for 15 min gave a vitreal/retinal level of GCV at 74 +/- 17 micrograms/ml at 2 hours as determined by HPLC. At 24 hours after iontophoresis the vitreal/retinal level was above therapeutic level at 4.2 +/- 0.6 micrograms/ml. At 72 hours, there was still detectable level in the vitreous/retina. Hence, transscleral iontophoresis is able to deliver effective dose of GCV into the vitreous. Multiple applications of iontophoresis should be examined as a possible means of CMV treatment.

Animals↗

A gene at 59 minutes on the Escherichia coli chromosome encodes a lipoprotein with unusual amino acid repeat sequences.

We report a 1.432-kb DNA sequence at 59 min on the Escherichia coli chromosome that connects the published sequences of the pcm gene for the isoaspartyl protein methyltransferase and that of the katF or rpoS (katF/rpoS) gene for a sigma factor involved in stationary-phase gene expression. Analysis of the DNA sequence reveals an open reading frame potentially encoding a polypeptide of 379 amino acids. The polypeptide sequence includes a consensus bacterial lipidation sequence present at residues 23 to 26 (Leu-Ala-Gly-Cys), four octapeptide proline- and glutamine-rich repeats of consensus sequence QQPQIQPV, and four heptapeptide threonine- and serine-rich repeats of consensus sequence PTA(S,T)TTE. The deduced amino acid sequence, especially in the C-terminal region, is similar to that of the Haemophilus somnus LppB lipoprotein outer membrane antigen (40% overall sequence identity; 77% identity in last 95 residues). The LppB lipoprotein binds Congo red dye and has been proposed to be a virulence determinant in H. somnus. Utilizing a plasmid construct with the E. coli gene under the control of a phage T7 promoter, we demonstrate the lipidation of this gene product by the incorporation of [3H]palmitic acid into a 42-kDa polypeptide. We also show that treatment of E. coli cells with globomycin, an inhibitor of the lipoprotein signal peptidase, results in the accumulation of a 46-kDa precursor. We thus designate the protein NlpD (new lipoprotein D). E. coli cells overexpressing NlpD bind Congo red dye, suggesting a common function with the H. somnus LppB protein. Disruption of the chromosomal E. coli nlpD gene by insertional mutagenesis results in decreased stationary-phase survival after 7 days.

Amino Acid Sequence↗

[Experimental study on oral candidiasis].

The successful rate in producing oral candidiasis of the animal is not high. By utilizing: (1) fresh cell-line isolated from the oral cavity of the patient suffering from oral candidiasis and passing twice in rabbits and re-isolated to increase its toxicity; and (2) expanding the area of examination, the authors successfully (100%) produced oral candidiasis in Wistar rats. The specimens are studied histologically on its nature of whether it is a precancerous lesion or not.

Animals↗

Beneficial effect of an ACTH-(4-9) analog (Org2766) in the treatment of experimental cortex lesion in rats.

This paper reports that lesions of the medial frontal cortex (MFC) caused behavioral deficit in rats and that this deficit could be attenuated by a well formulated treatment with Org2766, an analog of ACTH-(4-9). Wistar rats were distributed in 3 groups: MFC lesion treated with saline (M-N); MFC lesion treated with Org2766 (M-O); and sham-operation treated with saline (S). Repeated Org2766 or saline injection commenced from the day of surgery and lasted for 13 days. After surgery, the rats were trained in a passive avoidance task and then in an active avoidance task. MFC lesions were found to be strongly associated with behavioral deficits. The M-N group rats displayed poor retention of the passive avoidance response and showed much slower learning of the active avoidance task as compared to S group rats. The result showed that chronic Org2766 administration improved the behavioral performance of both tasks in MFC lesioned rats. The also revealed that the superoxide dismutase (SOD) activity was significantly increased in the M-O group as compared to the M-N group 15 days after surgery. The possible mechanisms related to the beneficial effect of Org2766 on cortex damage are discussed.

Adrenocorticotropic Hormone↗

Methylprednisolone therapy in laser injury of the retina.

The efficacy of methylprednisolone in argon-laser-induced retinal injury in primates was evaluated by clinical, histopathologic, and morphometric criteria. Methylprednisolone was given with a loading dose of 30 mg/kg followed by 5.4 mg/kg per hour in three different regimens: (1) starting 24 h before laser and continuing for 4 days; (2) starting immediately after laser and continuing for 4 days; and (3) starting immediately after laser and continuing for 8 h. Fundus photography, fluorescein angiography, and histologic examination showed significant beneficial effects of all three treatments compared to controls. Morphometrically, at the center of the lesion, the width of disrupted outer nuclear layer, the width of the affected RPE, and the percentage of residual photoreceptor nuclei confirmed the efficacies of treatment regimens 1 and 2, but not treatment regimen 3.

Animals↗

Genotoxic effects of subacute treatments with wood dust extracts on the nasal epithelium of rats: assessment by the micronucleus and 32P-postlabelling.

Human exposure to wood dust has been epidemiologically linked to a number of enhanced incidences of various neoplasias, including those of the nose. Among different suspected woods, long-term exposure to the dust of beech (Fagus silvatica) is strongly associated with development of these tumors. Experimentally, it has been shown that a simple alcoholic extract of beech wood dust is mutagenic toward some bacteria and tumorigenic on mouse skin. For the present study, different groups of male Wistar rats were treated with aqueous, ethanol, or methanol extract of untreated beech wood dust via nasal drip at three concentrations for 48 h. Animals were killed 24 h after the last treatment and the nasal tissues were isolated to be examined for possible induction of micronuclei and DNA adducts. A clear dose-dependent increase in the number of micronuclei (P < 0.01) was observed after treatment of rats with each alcoholic extract of wood dust. In contrast, no DNA adducts could be detected with these extracts using the 32P-postlabelling technique. No increased number of micronucleated cells was found with solvents alone or with aqueous extract of dust. These data might support the early hypothesis that wood dust per se contains some in vivo genotoxic and thus possibly carcinogenic components extractable by an alcohol.

Animals↗

Intelligence function in children with acute lymphoblastic leukemia after treatment.

In order to evaluate the effects of chemotherapy and central nervous system prophylaxis on the intelligence function of children with acute lymphoblastic leukemia (ALL), 147 patients were divided into 2 groups according to their different treatment regimens. Group A included 75 patients with ALL who were diagnosed and treated from 1981 to 1986. Group B included 72 ALL patients diagnosed and treated from 1988 to 1990. A control group included 73 healthy children who shared the same education and environmental background with the patients. Chinese revisions of Wechsler measures of intelligence quotient were used to estimate verbal IQ, performance IQ and total IQ in the 3 groups. American-produced Systate software was used to analyze the data. Results showed that IQ distribution, verbal IQ, performance IQ and total IQ of group A were obviously lower than those in the control group. Verbal IQ and total IQ of group B were also reduced. Significant differences of verbal IQ and performance IQ as well as total IQ were also found between groups A and B. Multiple regression analysis showed negative correlations between IQ and cranial irradiation dosage, times of intrathecal MTX and time after irradiation.

Antineoplastic Combined Chemotherapy Protocols↗

[Bioassay of Lyso-PAF platelet activating factor and it's role in bronchial asthma].

Plasma and lung homogenate lyso-PAF were measured by using the method of bioassay in guinea pigs with allergic asthma and asthmatic patients. The results showed that Lyso-PAF levels were increased significantly in asthmatics, and it was positively correlated with TXB2 levels. It is suggested that PAF plays an important role in bronchial asthma, which may be correlated with TXA2. The method of bioassay of Lyso-PAF is stable, reliable and special instruments are not needed.

Adolescent↗

Methylprednisolone ameliorates retinal photic injury in rats.

Since lipid peroxidation induced by oxygen free radicals is believed to play an important role in retinal photic injury and high doses of certain steroids have been demonstrated to inhibit lipid peroxidation, we evaluated the effect of high-dose methylprednisolone sodium succinate on retinal injury inflicted by green light (490 to 580 nm, 160 to 180 foot-candles [1721.6 to 1936.8 lux]) for 24 hours in 36 rats. Animals received either intraperitoneal injections of the drug (80 mg/kg twice daily) or saline as a control, twice daily for 2 days, commencing with light exposure. The retinal photic injury was assessed 6 hours, 6 days, and 14 days after light exposure by light and electron microscopy, morphometric study of outer nuclear layer thickness and count of subretinal macrophages, and spectrophotometric measurement of rhodopsin. A beneficial effect of high-dose methylprednisolone was observed in retinal photic injury.

Animals↗

Dexamethasone ameliorates retinal photic injury in albino rats.

The effect of dexamethasone in two regimens on retinal photic injury was studied in Lewis albino rats that were exposed to 24 hr of continuous green fluorescent light. Under regimen 1, dexamethasone was given at a daily dosage of 1 mg kg-1 for 8 days, starting 6 days before light exposure. Under regimen 2, dexamethasone was given at the same daily dosage for 3 days, started 1 day before light exposure. Pathologic study of the light-exposed retina, morphometric evaluation of the photoreceptor cell loss, cell counts of the macrophages in the subretinal space, and measurements of rhodopsin levels were undertaken in the dexamethasone-treated and control retinas at various times. The administration of dexamethasone in both regimens did not produce pathologic changes in the retina before light exposure, but rhodopsin levels were significantly lowered in both treated groups when compared to corresponding vehicle treated control animals. Under regimen 1, at 6 hr after light exposure, both the treated and the control groups showed comparable loss of photoreceptor cells, degeneration of the photoreceptor elements and retinal pigment epithelium, but a significantly lowered level of rhodopsin in the treated group was noted. At 6 days after exposure, the outer nuclear layer thickness, and the outer and inner segments showed significant preservation in the treated group. Also in the treated group, the number of macrophages was significantly reduced and the retinal pigment epithelial (RPE) vacuolation was markedly less. However, there was no difference in rhodopsin levels. At 14 days after exposure, the outer nuclear layer thickness and rhodopsin levels of the treated rats had significantly higher values than the controls. Under regimen 2, however, at 6 days after exposure, an ameliorative effect in the RPE was observed but there were no differences of rhodopsin levels, the outer nuclear thickness and number of macrophages between the treated and control groups. Regimen 1 was associated with a significantly higher retinal level of dexamethasone when compared with regimen 2. The ameliorative effect of dexamethasone on rat retinal photic injury may be through inhibition of lipid peroxidation, in which a high retinal level of the steroid is required.

Animals↗

Full-length cDNA sequence of dengue type 1 virus (Singapore strain S275/90).

The complete nucleotide sequence and the deduced amino acid sequence of the genome of dengue virus type 1 (Singapore strain S275/90) were determined from cDNA clones. The single-stranded, positive-sense RNA is 10,718 nucleotides in length and contains a single long open reading frame of 10,188 nucleotides encoding a polyprotein of 3396 amino acids. The genomic size and organization were found to be similar to that of other dengue virus serotypes. Both the nucleotide and deduced amino acid sequences were compared with the partial sequence of DEN1 (Nauru Island) and complete sequences of DNE2 (Jamaica), DEN3 (H87), and DEN4 (Dominica) virus genomes.

Amino Acid Sequence↗

Studies of fluoride retention by oral soft tissues after the application of home-use topical fluorides.

Previous studies have focused on enamel and plaque as the primary sites of fluoride (F) retention in the mouth. The present study was undertaken to evaluate the role of oral soft tissue as a site of F retention by comparing an edentulous subject panel (n = 9) with a fully dentate panel (n = 10). Unstimulated whole saliva samples were collected by having subjects pool saliva for two min. Samples were collected over a 24-hour period after application of a placebo dentifrice (PD; 0.4 ppm F), fluoride dentifrice (FD; 1100 ppm F), fluoride rinse (FR; 226 ppm F), or fluoride gel (FG; 5000 ppm F) delivered in custom trays. There was no statistically significant difference in salivary flow rate between the two panels for any of the treatments. The edentulous panel had higher salivary F levels than the dentate panel, which reached statistical significance (p less than 0.05) for the FD and FG treatments. In a separate study involving the same treatments, F levels at specific soft-tissue sites were measured over a one-hour period by use of absorbent discs placed in different soft-tissue areas of the mouth. The tongue and lower posterior vestibule retained the highest F levels, followed by the upper posterior buccal vestibule and upper anterior labial vestibule, with the lowest F levels retained in the lower anterior vestibule and the floor of the mouth. There was a strong-to-moderate correlation between whole saliva F concentration and F levels at specific soft-tissue sites. This study establishes the importance of oral soft tissue as the major site of F retention in the mouth.

Analysis of Variance↗

An improved intra-oral enamel demineralization test model for the study of dental caries.

The intra-oral enamel demineralization test (IEDT) was introduced by Brudevold et al. (1984). This caries model involves human subjects wearing palatal appliances each holding eight bovine enamel blocks covered by a bacterial cell layer prepared by the harvesting of cultures of Streptococcus mutants (test plaque). The original model used the iodide permeability test for assessment of the extent of demineralization of bovine enamel blocks resulting from acid production by the test plaque after dietary substrate challenge. The IEDT model has been expanded and improved by us in the following ways: (1) Based on encouraging findings from an in vitro study (Zero et al., 1990), the surface microhardness test has been adopted to measure the extent of demineralization occurring at three sites on the enamel blocks corresponding to an area over which the effective plaque thickness is 0.5, 1.5, and 2.5 mm; (2) intra-oral pH of the test plaque is measured by means of a Beetrode miniature pH electrode at baseline, then at five, 10, 15, 30, and 45 min after the start of a test; (3) plaque samples are collected at the end of a test and analyzed for organic acid content by means of HPLC; (4) the bacterial test challenge has been expanded to include different cariogenic bacteria which are grown under various growth conditions. The improved model has the capability of studying fundamental aspects of the caries process, namely, the relationships among dietary substrate challenge, plaque pH change, plaque organic acid profiles, microbial virulence properties, and enamel demineralization. Furthermore, the model has the potential for use in more applied research on caries-preventive agents such as fluoride.

Adult↗

Fluoride concentrations in plaque, whole saliva, and ductal saliva after application of home-use topical fluorides [published eerratum appears in J Dent Res 1993 Jan;72(1):87].

It is now well-accepted that the primary anti-caries activity of fluoride (F) is via topical action. The retention of F in the mouth after topical fluoride treatment is considered to be an important factor in the clinical efficacy of F. The purpose of this study was to evaluate F levels in ductal saliva, whole saliva, and pooled plaque after treatment with topical F agents intended for home use. Ten consenting adults, mean (SD) age 31.0 (8.2) years, participated in all aspects of the study. Two days before each test, subjects received a professional tooth cleaning and subsequently abstained from all oral hygiene procedures to permit plaque to accumulate, and from the use of F-containing dental products. Treatments consisted of a placebo dentifrice (PD), fluoride dentifrice (FD; 0.24% NaF), fluoride rinse (FR; 0.05% NaF), and fluoride gel (FG; 1.1% NaF). Unstimulated whole saliva and pooled plaque were sampled at multiple points over a 24-hour period. In a separate experimental series, stimulated parotid saliva was sampled over a two-hour period after treatment. Fluoride levels generally followed the same pattern in whole saliva and pooled plaque samples, with FG > FR > FD > PD. Night-time F application resulted in prolonged F retention in whole saliva but not in plaque. Fluoride levels in parotid saliva were only slightly higher after F treatment and returned to baseline levels within two h. The results of this study indicate that the method of F delivery, the F concentration of the agent, and the time of application (daytime vs. night-time) are important factors influencing F levels in the mouth.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

[Mutagenesis study of ethylmethane sulfonate with shuttle vector plasmid pZ189].

Shuttle vector plasmid pZ189 was used as a molecular tool and the SupF inserted in the plasmid was worked as a target gene for mutagenesis study. The host cells (E. coli MBM 7070) with pZ189 were treated with ethylmethane sulfonate (EMS) and plated on the selective media containing X-gal (5-bromo-4-chloro-3-indolyl-beta-D-galactoside) and IPTG (isopropyl-beta-D-thiogalactoside). The SupF and the LacZ amber mutant carried by the host cells complemented each other and thus made the colonies blue on the selective media. However the colonies derived from the SupF mutants changed the colour from blue to white. The mutant frequencies in a series of experiments with different concentrations of EMS were estimated. Furthermore, the DNA isolated from 5 SupF mutants was digested with restruiction enzyme Hha I. It suggests that the 214 bp Hha I fragments containing mutant SupF could be distinguished from their wild type counterparts by temperature-gradient gel electrophoresis under optimal conditions.

Electrophoresis, Polyacrylamide Gel↗