Search PubMed⌕ Search

Biomedical subjects

S Y Wu

Publications and source records attributed to S Y Wu.

At least 37 records · Page 2Linked to original sources
PubMed2001

Patterns of adherence to diabetes vision care guidelines: baseline findings from the Diabetic Retinopathy Awareness Program.

OBJECTIVES: (1) To describe baseline patterns of adherence to American Diabetes Association and American Academy of Ophthalmology vision care guidelines for diabetes in the Diabetic Retinopathy Awareness Program, and (2) to evaluate factors associated with nonadherence. This paper describes the baseline characteristics of a population enrolled in a prospective, randomized clinical trial. DESIGN: Cross-sectional study. PARTICIPANTS: Between October 1993 and May 1994, the study identified 2308 persons with diabetes, 18 years of age or older, who were residents of Suffolk County, New York, via a multimedia community-wide recruitment campaign. INTERVENTION AND METHODS: Eligibility for the trial was determined during a 20-minute phone interview, which included questions about vision care practices; diabetes management; and knowledge, attitudes, and beliefs about diabetes, vision, and diabetic retinopathy. This paper describes these patient characteristics at baseline. Eligible patients would be randomized subsequently to a 2-year diabetes educational intervention arm, which included mailed packets and newsletters focused on vision care, or to a control nonintervention arm. MAIN OUTCOME MEASURE: Nonadherence to guidelines at baseline was defined as the absence of a dilated eye examination during the year before recruitment into the study. RESULTS: Of the 2308 persons interviewed, 813 (35%) did not follow the vision care guidelines; two thirds of this group reported no eye examination in the year before the interview, and one third had an undilated examination. Ophthalmologists performed 49% of the examinations in the nonadherent group, versus 86% in the adherent group. In logistic regression analyses, factors related to nonadherence were: younger age (odds ratio [OR] = 0.97), type 2 diabetes with or without insulin use (OR = 1.62 and 1.99, respectively), shorter diabetes duration (OR = 0.97), last eye examination performed by an optometrist (OR = 5.32) or other nonophthalmologist (OR = 4.29), less practical knowledge about diabetes (OR = 1.57), and no prior formal diabetes education (OR = 1.30). CONCLUSIONS: Within this population, more than one third of participants had not been following vision care guidelines. Nonadherence was linked to several potentially modifiable factors; changes in these factors could enhance the early detection of diabetic retinopathy.

Adolescent↗
PubMed2001

Patterns of open-angle glaucoma in the Barbados Family Study.

OBJECTIVE: To describe the Barbados Family Study of open-angle glaucoma (OAG) and present risk factors for OAG in siblings of study probands. DESIGN: Observational study of families of probands with OAG. PARTICIPANTS: Two hundred thirty probands and 1056 relatives (from 207 families). METHODS: Probands and their family members underwent standardized examinations, including automated perimetry, applanation tonometry, ophthalmologic evaluation, fundus photography, blood pressure, interview, and genotyping. Generalized estimation equation methods were used to evaluate risk factors. MAIN OUTCOME MEASURES: Presence of OAG in the relatives, as defined by both visual field and optic disc findings, after ophthalmologic exclusion of other causes. RESULTS: The median ages of probands and relatives were 68 and 47 years, respectively. In the 207 families, 29% of the probands had one relative with OAG and 10% had two or more relatives affected. Of the 1056 family members, 10% had OAG, 13% had suspect OAG, and 6% had ocular hypertension. One fifth of the 338 siblings had OAG (n = 67); they tended to be older and more often were male. Multivariate comparisons between siblings with and without OAG found that age, higher intraocular pressure (IOP), myopia, and lower diastolic blood pressure-IOP differences were related to OAG, whereas hypertension and diabetes were not. CONCLUSIONS: Based on standardized protocols and examinations, approximately one quarter of the relatives had OAG or suspected OAG, despite their relatively young age. Risk factors for OAG in siblings were similar to risk factors in unrelated individuals. Analyses are ongoing to determine OAG inheritance and to localize potential gene(s) involved.

Adult↗
PubMed2001

Prevalence and causes of visual impairment in The Barbados Eye Study.

OBJECTIVE: To determine the prevalence and causes of low vision and blindness in a predominantly black population. DESIGN: Population-based prevalence study of a simple random sample of Barbados-born citizens aged 40 to 84 years. PARTICIPANTS: Four thousand seven hundred nine persons (84% participation). METHODS: The standardized protocol included best-corrected visual acuity (with a Ferris-Bailey chart), automated perimetry, lens gradings (LOCS II), and an interview. Participants with visual acuity of worse than 20/30, other positive findings, and a 10% sample also had an ophthalmologic examination that evaluated the cause and extent of vision loss (resulting from that cause), if any. MAIN OUTCOME MEASURES: Low vision and blindness were defined as visual acuity in the better eye between 6/18 and 6/120 and visual acuity worse than 6/120, respectively (World Health Organization [WHO] criteria). RESULTS: Of the 4631 participants with complete examinations, 4314 (93%) reported their race as black, 184 (4%) reported their race as mixed (black and white), and 133 (3%) reported their race as white or other. Low vision was found in 5.9% of the black, 2.7% of the mixed, and 3.0% of white or other participants. Bilateral blindness was similar for black and mixed race participants (1.7% and 1.6%, respectively) and was not found in whites. Among black and mixed participants, the prevalence of low vision increased with age (from 0.3% at 40-49 years to 26.8% at 80 years or older). The prevalence of blindness was higher (P < 0.001) for men than women at each age group (0.5% versus 0.3% at ages 40-49 and 10.9% versus 7.3% at 80 years or more). Sixty percent of blindness was due to open-angle glaucoma and age-related cataract, each accounting for more than one fourth of cases. Other major causes were optic atrophy or neuropathy and macular and other retinal diseases. Few cases of blindness were due to diabetic retinopathy (1.4%), and none were due to age-related macular degeneration. CONCLUSIONS: Using the WHO criteria, prevalence of visual impairment was high in this African-origin population, particularly at older ages. Most blindness was due to open-angle glaucoma and cataract, with open-angle glaucoma causing a higher proportion of blindness than previously reported. The increased prevalence of blindness in men may be due to the increased male prevalence of glaucoma in this population and warrants further investigation. Results underline the need for blindness prevention programs, with emphasis on effective treatment of age-related cataract and enhancing strategies for early detection and treatment of open-angle glaucoma.

Adult↗
PubMed2001

Reconstitution of recombinant TFIIH that can mediate activator-dependent transcription.

BACKGROUND: TFIIH is one of the general transcription factors required for accurate transcription of protein-coding genes by RNA polymerase II. TFIIH has helicase and kinase activities, plays a role in promoter opening and promoter escape, and is also implicated in efficient activator-dependent transcription. RESULTS: We have established a reconstitution system of recombinant TFIIH using a three-virus baculovirus expression system. The recombinant TFIIH was active in CTD kinase and DNA helicase assays, and showed both basal and activator-dependent transcriptional activities that were indistinguishable from those of HeLa cell-derived TFIIH. Further analyses using recombinant TFIIH confirmed a critical role of TFIIH in activator-dependent transcription. The dose response of TFIIH in activator-dependent transcription suggested that mere recruitment of TFIIH is not sufficient for transcriptional activation. The sensitivity of activator-dependent transcription to nonhydrolysable ATP analogues indicated the importance of the enzymatic activities of TFIIH in transcriptional activation. CONCLUSIONS: Our results raise a possibility that transcriptional activation by GAL4-VP16 requires enzymatic activities. Recombinant TFIIH reconstituted from this baculovirus system should be useful for analysis of the mechanisms of activation by GAL4-VP16.

Adenosine Triphosphate↗
PubMed2001

The correlation of cytokine levels with body weight after megestrol acetate treatment in geriatric patients.

BACKGROUND: Cachexia is associated with elevated levels of cytokines in cancer and human immunodeficiency virus patients. Studies in cancer and acquired immunodeficiency syndrome patients showed that treatment with megestrol acetate (MA) is associated with improvement in appetite and weight gain. Reduction in the levels of cytokines is associated with weight gain in laboratory animals with cancer. This study evaluates the correlation between changes in cytokine (or their receptor) levels and weight following MA treatment in geriatric weight-loss patients. METHODS: Veterans Administration Medical Center nursing home patients (N = 69) with a weight loss of > or =5% of usual body weight over the past 3 months or body weight 20% below their ideal body weight participated in a 12-week, randomized, double-blind, placebo-controlled trial, with an additional 13-week follow-up period. Patients were randomly assigned to receive a placebo or MA oral suspension of 800 mg/d for 12 weeks. Levels of the following cytokines (or their receptors) were measured at baseline and after 12 weeks of treatment: tumor necrosis factor soluble receptor (TNFR) subunits. TNFR-p55 and TNFR-p75: interleukin 6 (IL-6); and the soluble interleukin-2 receptor (sIL-2R). The subjects' weight and body composition were measured at the start of the study. Weight and mortality were followed up for another 13 weeks after discontinuing the MA study drug. RESULTS: Elevated levels of IL-6 in almost all geriatric cachexic patients, compared with normal volunteers (mean, <4.6 pg/ml). were noted at baseline. At 12 weeks after the study drug treatment, there was a decrease in cytokine levels (or their receptors) in the MA group (mean change in IL-6, 3.63+/-6.62 pg/ml; TNFR-p55, -0.06+/-0.11 ng/ml; TNFR-p75. -0.01+/-0.29 ng/ml; and sIL-2R, 0.08+/-0.07 ng/ml) and the placebo group (mean change in IL-6, -2.08+/-3.92 pg/ml; TNFR-p55, -0.02+/-0.08 ng/ml; TNFR-p75, -0.20+/-0.18 ng/ml; and sIL-2R, 0.02+/-0.03 ng/ml). Although the change in cytokine levels was not statistically significant between the two groups, significant negative correlation (p < .05) was found. For example, increased weight correlated with decreased sIL-2R levels (r = .36) and TNFR-p75 (r = -.31; fat-free mass (FFM) gain and reduction of sIL-2R (r = -.39), TNFR-p75 (r = -.30). There was a significant correlation between weight gain and reduction of TNFR-p75 (r = .54), TNFR-p55 (r- = .47), and sIL-2R (r = -.53); FFM gain and reduction of sIL-2R (r = -.59), TNFR-p75 (r = -.41), TNFR-p55 (r = -.42); and fat gain and reduction of TNFR-p75 (r = -.41) in the MA group (p < .05), but not in the placebo group. CONCLUSIONS: Although there was no significant change in cytokine levels between the two groups, the reduction in cytokine levels after MA treatment correlated with improvement in weight, fat mass, and FFM at 12 weeks.

Aged↗
PubMed2001

3,3'-Diiodothyronine sulfate excretion in maternal urine reflects fetal thyroid function in sheep.

We have shown that there is significant fetal-to-maternal transfer of sulfated metabolites of thyroid hormone after fetal infusion of a pharmacologic amount of 3,3',5-triiodothyronine (T(3)) or sulfated T(3) in late pregnancy in sheep (Am J Physiol 277:E915, 1999). The transferred iodothyronine sulfoconjugate, i.e. 3,3'-diiodothyronine sulfate (T(2)S), of fetal origin appears in maternal sheep urine. The present study was carried out to assess the contribution of T(2)S of fetal origin to the urinary pool in ewes. Eighteen date-bred ewes (mean gestational age of 115 d) and their twin fetuses were divided into four groups. In group I (control, n = 5), both ewes (M) and their fetuses (F) were sham operated for thyroidectomy (Tx). In group II, the ewes (MTx, n = 4) and, in group III, the fetuses (FTx, n = 4) were subjected to Tx. In group IV (MTx.FTx, n = 5), both the ewe and fetus had Tx. After 10-12 d, fetal and/or maternal hypothyroidism were confirmed by serum thyroxine (<15 nmol/L) measurements. In addition, we infused radioactive T(3) without disturbing the T(3) pool in three singleton near-term fetuses and assessed the amount of radioactive iodothyronine that appeared in maternal urine (MU). After infusing [(125)I-3'],3,5-T(3) via fetal vein to the near-term normal fetuses, radioactive T(2)S was identified as the major metabolite in MU by HPLC and T(2)S-specific antibody. MU T(2)S excretion (pmol/mmol creatinine) was significantly reduced by FTx and MTx.FTx but not by MTx. In addition, positive correlations (p < 0.01) were found between MU T(2)S excretion and fetal serum thyroxine and T(3) concentrations but not with maternal serum thyroxine or T(3) levels. T(2)S of fetal origin contributes significantly to the MU pool.

Animals↗
PubMed2001

Incidence of open-angle glaucoma: the Barbados Eye Studies. The Barbados Eye Studies Group.

OBJECTIVE: To measure the 4-year risk of open-angle glaucoma (OAG) in a black population. DESIGN: Population-based cohort study with 4 years of follow-up. SETTING: Simple random sample of residents of Barbados, West Indies, aged 40 years or older. PARTICIPANTS: A total of 3427 members of the cohort (85% of those eligible). MAIN OUTCOME MEASURE: Development of glaucoma visual field defects and optic disc damage, confirmed by automated perimetry, independent fundus photographic gradings, and standardized ophthalmologic examinations. RESULTS: The 4-year risk of OAG in black participants was 2.2% (95% confidence interval, 1.7%-2.8%), based on 67 newly developed cases of OAG. Incidence rates increased from 1.2% at ages 40 to 49 years to 4.2% at ages of 70 years or more, tending to be higher in men than women (2.7% vs 1.9%). About half of the incident cases were undiagnosed previously, and the rest were receiving OAG treatment. Of the 67 new cases of OAG, 32 had intraocular pressure of 21 mm Hg or less at baseline (1.2% incidence) and 35 had higher pressures (9% incidence). Risk was highest among persons classified as having suspect OAG at baseline (26.1%), followed by those with ocular hypertension (4.9%) and lowest in the remaining population (0.8%). CONCLUSIONS: This longitudinal study provides new information on OAG risk, as well as the first incidence measurement in a black population. Although intraocular pressure increased risk, about half of the new cases had baseline pressures of 21 mm Hg or less. Results substantiate the high OAG risk in the population of African origin, especially in older adults; the relative role of intraocular pressure; and the considerable underdetection of new disease after 4 years of follow-up.

Adult↗
PubMed2001

Hepatitis A virus infection in Taipei in 1999.

BACKGROUND AND PURPOSE: Hepatitis A is a disease that is heavily affected by sanitation status. Hepatitis A is much less prevalent compared with decades ago in Taiwan, as in many rapidly developing regions. Hepatitis A vaccine is still self-paid under the National Health Insurance program and is still not widely utilized by the general public in Taiwan. This seroepidemiologic study evaluated the prevalence of antihepatitis A virus (anti-HAV) seropositivity in Taipei in 1999. METHODS: A total of 1017 serum samples from healthy inhabitants in Taipei were examined for anti-HAV antibody by qualitative enzyme immunoassay. RESULTS: The overall seroprevalence rate was 25.2% (255/1013) in the nonvaccinated population. The seropositivity rate for anti-HAV antibody among children younger than 12 months old was 23.3%. The rates dropped to between 1% and 4.8% among subjects between 1 and 20 years of age. A markedly higher rate of 40% was observed in subjects aged between 20 and 30 years. The seropositivity rate in subjects aged 31 to 50 was 80%. More than 90% of subjects older than 50 years were seropositive. The vaccination rate was low (0.5%). CONCLUSION: Our findings indicate that Taipei is an area of intermediate endemicity for hepatitis A virus. To achieve better herd immunity, a more active approach to the adoption of hepatitis A vaccine is warranted.

Adolescent↗
PubMed2001

[Primary analysis and sequencing the hemolytic relative gene of Edwardsiella tarda].

Hemolysin is an important pathogenic agent of Edwardsiella tarda (ET). A fragment containing hemolytic gene, of which sequence is 4,264 bp, was cloned from ET-12 chromosome by shutgun method. It has no similarity to the hemolysin genes reported, of which 424 bp of open read frame (ORF) 3 has 68% similarity to the hemolysin regulatory gene of S. typhi (slyA). The subclone which has complete ORF3 was hemolytic, and the other of which Kanamycin gene was inserted in Pvu I of ORF3 was no hemolytic. It proved the gene was in chromosomes of ET-12 and the other ET strains by hybridization in situ and Southern blot. As recombination plasmid with the gene entered nonhemolytic ET by electroporation, no hemolytic phenomenon was observed. Conclusion was that the gene wasn't the hemolysin gene and was hemolytic relative gene.

Amino Acid Sequence↗
PubMed2000

Linkage analysis with candidate genes: the Taiwan young-onset hypertension genetic study.

A genetic linkage study of young-onset hypertension was performed on data from 59 nucleus families of Han Chinese residing in Taiwan. Thirty seven microsatellite markers near 18 hypertension candidate genes were genotyped. In a nonparametric identity-by-descent sibpair analysis, a positive linkage signal (defined as P<0.05) was found for four microsatellite markers, viz., D1S1612 (P=0.0162), D1S547 (P=0.0263), D8S 1145 (P= 0.0284), and D17S2193 (P=0.0256), which were located near genes for atrial natriuretic peptide (NPPA)/glucose transporter 5 (SLC2A5), angiotensinogen (AGT), lipoprotein lipase (LPL), and angiotensin-conveting enzyme (DCP1), respectively. Marker D5S1480 located near beta-2-adrenergic receptor (ADRB2) had a borderline P value (P=0.0785) for the positive signal. Comprehensive genotyping with further markers in these regions is underway to confirm whether these genes are linked to young-onset hypertension.

Adult↗
PubMed2000

Glaucoma and myopia.

Explore the source record for details and available documents.

Glaucoma, Open-Angle↗
PubMed2000

Incidence and progression of lens opacities in the Barbados Eye Studies.

OBJECTIVE: To provide 4-year cumulative incidence and progression rates of age-related lens opacities in a population > or =40 years of age, which is mainly of African origin. DESIGN: Cohort study that reexamined surviving members of the population-based Barbados Eye Study 4 years after baseline. PARTICIPANTS: Three thousand four hundred twenty-seven members of the Barbados Eye Study cohort (85% of those eligible). MAIN OUTCOME MEASURES: The Lens Opacities Classification System II (LOCS II) was used at the slit lamp. Cumulative incidence was defined as the development of any nuclear, cortical or posterior subcapsular (PSC) opacities (LOCS II scores > or =2) among persons without that opacity type at baseline. Cumulative progression was defined by at least two-step increases in scores among persons with preexisting lens opacities. RESULTS: The incidence of cortical opacities was about five times greater in black than white participants (age-gender adjusted relative risk = 4.7; 95% confidence interval: 1.9-11.4). In the black population, the 4-year incidence rates were 22.2% (20.4%-24.0%) for any cortical, 9.2% (8.2%-10.4%) for any nuclear, and 3.3% (2.7%-4.0%) for any PSC opacities; rates increased greatly with age. Four-year progression rates were 12.5% for cortical, 3.6% for nuclear, and 23.0% for PSC opacities, without consistent pattern by age. Women had a greater risk of cortical and nuclear opacities (P<0.05) than men and greater progression of nuclear opacities. The presence of PSC opacities at baseline seemed to at least double the incidence and progression rates of other opacities. In persons initially opacity free, single cortical opacities were the predominant type to develop at followup. Visual acuity loss frequently accompanied incident opacities. CONCLUSIONS: This longitudinal study provides new population-based data on the natural history of lens opacities. Incidence and progression of opacities, especially of cortical opacities, were high. After 4 years of followup, 1 in 4 to 5 participants developed cortical opacities, 1 in 11 developed nuclear opacities, and 1 in 30 developed PSC opacities. The information obtained attests to the public health impact of age-related cataract, as well as its extent, in this and similar black populations.

Adult↗
PubMed2000

Recruiting participants for community-based research: the Diabetic Retinopathy Awareness Program.

PURPOSE: Recruiting participants is a major challenge for population studies. We present the recruitment methods followed by the Diabetic Retinopathy Awareness Program (DRAP), a community-based, randomized, masked, controlled trial to meet and exceed its sample size goals. METHODS: A county-wide multi-media promotional campaign to recruit and enroll participants in the trial was planned and executed from October 1993 through April 1994, with the assistance of the local news media and community and professional groups. A toll-free 800 number recruitment line was established, and postage-paid recruitment postcards distributed. The trial was designed to examine whether a mailed educational intervention could increase compliance with vision care guidelines among persons with diabetes in the community. RESULTS: A total of 2308 persons with diabetes were interviewed for eligibility and 813 enrolled in the intervention trial, exceeding the original recruitment goals of 1800 and 600, respectively. Those who completed the enrollment interview reflected county demographics. During recruitment, newspaper, television and radio stories featured the study; pharmacies and physician offices displayed study materials; public service announcements appeared in local print and broadcast media. The largest single recruitment response was a local television news report, followed by a newspaper story. CONCLUSIONS: These experiences substantiate the need for a comprehensive coordinated approach, using planned multiple sources, to achieve recruitment success. By engaging the lay and professional communities along with the media, recruitment costs can be kept to a minimum. Participant costs can be minimized by employing a toll-free number and eliminating study participant travel, thus allowing for inclusion of traditionally underserved populations. This approach is applicable to other studies, where community-based participation is desired.

Adolescent↗
PubMed2000

Improvement in quality-of-life measures and stimulation of weight gain after treatment with megestrol acetate oral suspension in geriatric cachexia: results of a double-blind, placebo-controlled study.

BACKGROUND: Weight loss among older patients is a severe problem, associated with an increased incidence of infections, decubiti, and death. Megestrol acetate (MA) causes weight gain in cachectic cancer and AIDS patients, but its effects in older cachectic patients are unknown. OBJECTIVE: To compare the effects of MA oral suspension (O.S.), 800 mg/day, versus placebo on weight in geriatric nursing home patients with weight loss or low body weight. DESIGN: Twelve-week, randomized, double-blind, placebo-controlled trial with a 13-week follow-up period. SETTING: Veterans Administration Medical Center (VMAC) nursing home. PATIENTS: Nursing home patients with weight loss of > or =5% of usual body weight over the past 3 months, or body weight 20% below their ideal body weight. INTERVENTIONS: Patients were randomly assigned to receive placebo or MA 800 mg/day for 12 weeks and were then followed for 13 weeks off treatment. MEASUREMENTS: Primary outcome was measured by weight and appetite change. Secondary outcome measures included sense of well-being, enjoyment of life, change in depression scale, laboratory nutrition parameters, energy intake counts, body composition, and adverse events. RESULTS: At 12 weeks there were no significant differences in weight gain between treatment groups, whereas MA-treated patients reported significantly greater improvement in appetite, enjoyment of life, and well-being. Body composition was not statistically different between the two groups. At Week 25 (3 months after treatment), 61.9% of MA-treated patients had gained > or =1.82 kg (4 lbs) compared to 21.7% of placebo patients. CONCLUSIONS: In geriatric patients with weight loss or low body weight MA improves appetite and well-being after 12 weeks of treatment. During the 3 months of MA treatment, there was no statistically significant weight gain (> or =4 lbs). Three months after treatment, weight gain (> or =4 lbs) was significantly increased in MA-treated patients.

Administration, Oral↗
PubMed2000

Origin of hepatitis delta virus mRNA.

Hepatitis delta virus (HDV) is unique relative to all known animal viruses, especially in terms of its ability to redirect host RNA polymerase(s) to transcribe its 1,679-nucleotide (nt) circular RNA genome. During replication there accumulates not only more molecules of the genome but also its exact complement, the antigenome. In addition, there are relatively smaller amounts of an 800-nt RNA of antigenomic polarity that is polyadenylated and considered to act as mRNA for translation of the single and essential HDV protein, the delta antigen. Characterization of this mRNA could provide insights into the in vivo mechanism of HDV RNA-directed RNA transcription and processing. Previously, we showed that the 5' end of this RNA was located in the majority of species, at nt 1630. The present studies show that (i) at least some of this RNA, as extracted from the liver of an HDV-infected woodchuck, behaved as if it contained a 5'-cap structure; (ii) in the infected liver there were additional polyadenylated antigenomic HDV RNA species with 5' ends located at least 202 nt and even 335 nt beyond the nt 1630 site, (iii) the 5' end at nt 1630 was not detected in transfected cells, following DNA-directed HDV RNA transcription, in the absence of genome replication, and (iv) nevertheless, using in vitro transcription with purified human RNA polymerase II holoenzyme and genomic RNA template, we did not detect initiation of template-dependent RNA synthesis; we observed only low levels of 3'-end addition to the template. These new findings support the interpretation that the 5' end detected at nt 1630 during HDV replication represents a specific site for the initiation of an RNA-directed RNA synthesis, which is then modified by capping.

5' Untranslated Regions↗
PubMed2000

Alleviation of human papillomavirus E2-mediated transcriptional repression via formation of a TATA binding protein (or TFIID)-TFIIB-RNA polymerase II-TFIIF preinitiation complex.

Transcription in human papillomaviruses (HPVs) is mainly regulated by cellular transcription factors and virus-encoded E2 proteins that act as sequence-specific DNA-binding proteins. Although the functions of E2 as a transcriptional activator and a repressor have been well documented, the role of cellular factors involved in E2-mediated regulation of the HPV promoters and the mechanism by which E2 modulates viral gene expression remain unclear. Using reconstituted cell-free transcription systems, we found that cellular enhancer-binding factors and general cofactors, such as TAF(II)s, TFIIA, Mediator, and PC4, are not required for E2-mediated repression. Unlike other transcriptional repressors that function through recruitment of histone deacetylase or corepressor complexes, HPV E2 is able to directly target components of the general transcription machinery to exert its repressor activity on the natural HPV E6 promoter. Interestingly, preincubation of TATA binding protein (TBP) or TFIID with HPV template is not sufficient to overcome E2-mediated repression, which can be alleviated only via formation of a minimal TBP (or TFIID)-TFIIB-RNA polymerase II-TFIIF preinitiation complex. Our data therefore indicate that E2 does not simply work by displacing TBP or TFIID from binding to the adjacent TATA box. Instead, E2 appears to function as an active repressor that directly inhibits HPV transcription at steps after TATA recognition by TBP or TFIID.

Base Sequence↗
PubMed2000

Nociceptin inhibits rat sympathetic preganglionic neurons in situ and in vitro.

In vitro and in situ experiments were conducted to evaluate the hypothesis that the nonclassical opioid peptide nociceptin acting on sympathetic preganglionic neurons (SPNs) inhibits spinal sympathetic outflow. First, whole cell patch recordings were made from antidromically identified SPNs from immature (12-16 day old) rat spinal cord slices. Nociceptin (0.1, 0.3, and 1 microM) concentration dependently suppressed the excitatory postsynaptic potentials (EPSPs) evoked by focal stimulation and hyperpolarized a population of SPNs; these effects were naloxone insensitive. L-Glutamate-induced depolarizations were not significantly changed by nociceptin. Results from this series of experiments indicate that nociceptin inhibits the activity of SPNs by either a presynaptic or postsynaptic site of action, whereby the peptide reduces, respectively, the amplitude of EPSPs or the excitability of SPNs. Second, intrathecal injection of nociceptin (3, 10, and 30 nmol) to urethan-anesthetized rats dose dependently reduced the mean arterial pressure and heart rate; these effects were not prevented by prior intravenous administration of naloxone (1 mg/kg). Physiological saline given intrathecally was without appreciable effects. These results, together with earlier observations of the detection of nociceptin-immunoreactive nerve fibers and nociceptin receptor immunoreactivity in the rat intermediolateral cell column, raise the possibility that the opioid peptide, which may be released endogenously, reduces spinal sympathetic outflow by depressing the activity of SPNs.

Animals↗