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Biomedical subjects

H Jiang

Publications and source records attributed to H Jiang.

At least 55 records · Page 3Linked to original sources
PubMed2004

Scd1ab-Xyk: a new asebia allele characterized by a CCC trinucleotide insertion in exon 5 of the stearoyl-CoA desaturase 1 gene in mouse.

We describe here a spontaneous, autosomal recessive mutant mouse suffering from skin and hair defects, which arose in the outbred Kunming strain. By haplotype analysis and direct sequencing of PCR products, we show that this mutation is a new allele of the asebia locus with a naturally occurring mutation in the Scd1 gene (a CCC insertion at nucleotide position 835 in exon 5), which codes for stearoyl-CoA desaturase 1. This mutation introduces an extra proline residue at position 279 in the Scd1 protein. The mutant mice, originally designated km/km but now assigned the name Scd1ab-Xyk (hereafter abbreviated as abXyk/abXyk), have a similar gross and histological phenotype to that reported for previously characterized allelic asebia mutations (Scd1ab, Scd1abJ, Scd1ab2J, and Scd1tm1Ntam). Histological analysis showed they were also characterized by hypoplasic sebaceous glands and abnormal hair follicles. In a cross between Kunming- abXyk/abXyk and ABJ/Le-abJ/abJ mice, all the progeny showed the same phenotype, indicating that the two mutations were non-complementing and therefore allelic. Comparisons with the other four allelic mutants indicate that the Scd1ab-Xyk mutation causes the mildest change in Scd1 function. This new mouse mutant is a good model not only for the study of scarring alopecias in humans, which are characterized by hypoplasic sebaceous glands, but also for studying the structure and function of the Scd1 protein.

Alleles↗
PubMed2004

Epoxyeicosatrienoic acids mediate adenosine-induced vasodilation in rat preglomerular microvessels (PGMV) via A2A receptors.

Activation of rat adenosine2A receptors (A2A R) dilates preglomerular microvessels (PGMV), an effect mediated by epoxyeicosatrienoic acids (EETs). Incubation of PGMV with a selective A2A R agonist, 2-p-(2-carboxyethyl) phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS 21680; 100 microM), increased isolated PGMV EET levels to 7.57+/-1.53 ng mg-1 protein from 1.06+/-0.22 ng mg-1 protein in controls (P<0.05), without affecting hydroxyeicosatetraenoic acid (HETE) levels (10.8+/-0.69 vs 11.02+/-0.74 ng mg-1 protein). CGS 21680-stimulated EETs was abolished by preincubation with an A2A R antagonist, 4-(2-[7-amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM241385) (100 microM). A selective epoxygenase inhibitor, methylsulfonyl-propargyloxyphenylhexanamide (MS-PPOH; 12 microM) prevented CGS 21680-induced increase in EETs, indicating inhibition of de novo synthesis of EETs. In pressurized (80 mmHg) renal arcuate arteries (110-130 microm) preconstricted with phenylephrine (20 nM), superfusion with CGS 21680 (0.01-10 microM) increased the internal diameter (i.d.) concentration-dependently; vasodilation was independent of nitric oxide and cyclooxygenase activity. CGS 21680 (10 microM) increased i.d. by 32+/-6 microm; vasodilation was prevented by inhibition of EET synthesis with MS-PPOH. Addition of 3 nM 5,6-EET, 8,9-EET and 11,12-EET increased i.d. by 53+/-9, 17+/-4 and 53+/-5 microm, respectively, whereas 14,15-EET was inactive. The responses to 5,6-EET were, however, significantly inhibited by indomethacin. We conclude that 11,12-EET is the likely mediator of A2A R-induced dilation of rat PGMV. Activation of A2A R coupled to de novo EET stimulation may represent an important mechanism in regulating preglomerular microvascular tone. British Journal of Pharmacology (2004) 141, 441-448. doi:10.1038/sj.bjp.0705640

8,11,14-Eicosatrienoic Acid↗
PubMed2004

Frequency and quantity of the parvovirus B19 genome in endomyocardial biopsies from patients with suspected myocarditis or idiopathic left ventricular dysfunction.

Parvovirus B19 (PB19) has been identified as a possible cause of myocarditis and heart failure in both children and adult patients. This study used real time PCR analysis, to determine the frequency and to quantify PB19 viral genomes in endomyocardial tissue samples from 80 adult patients with clinically suspected myocarditis or idiopathic left ventricular dysfunction and from 36 controls. Histological (Dallas classification) and immunohistological analyses were performed to detect myocardial inflammation in the endomyocardial biopsies.PB19 genomic DNA was found in nine of 80 patients (11.2%), 4 out of 31 (12.9%) patients with inflammatory infiltrates detected via immunohistological methods and 5 out of 49 (10.2%) patients with left ventricular dysfunction without myocardial inflammation. The copy numbers for PB19 DNA ranged between 30 and 3900 per microg of cellular DNA. Four patients with clinically suspected myocarditis had copy numbers for PB19 DNA of 70, 740, 3400 and 3900, respectively, per microg of cellular DNA in the endomyocardial biopsy. Five patients with idiopathic left ventricular dysfunction had copy numbers for PB19 DNA of 30, 38, 52, 58 and 90, respectively, per microg of cellular DNA in the endomyocardial biopsy. The amplicon of one of the nine positive PCR fragment was sequenced and was found to be fully identical in the highly conserved sequence of published Parvovirus B19 VP1/VP2 genes (NCBI gene bank). In all patients, acute myocarditis was excluded according to the Dallas classification. All biopsies of 36 controls with no history of myocarditis or recent viral infection were negative for myocardial inflammation and parvovirus B19 genomes. In summary, Parvovirus B19 DNA is present within the myocardium of patients with suspected myocarditis and idiopathic left ventricular dysfunction and can be detected and quantified in endomyocardial specimens via real time PCR.

Adult↗
PubMed2004

Ki-67-directed antisense therapy in an orthotopic renal cell carcinoma model.

PURPOSE: The Ki-67 antigen is only present in proliferating cells. We have shown previously that phosphorothioate-modified antisense oligonucleotides (ON) against this antigen are potent antitumoral agents in bladder and prostate cancer-derived cells. Since ON are known to accumulate in vivo in the kidney, high local effectivity may be expected. Here, we evaluated and characterized antitumoral effects in an orthotopic renal cell cancer (RENCA) model. MATERIAL AND METHODS: RENCA cells were incubated with antisense and control ON in the presence of a cationic lipid. Uptake studies were performed with FITC-labeled ON. Ki-67 protein analysis after ON treatment was performed by immunohistochemical staining. For animal studies, 1 x 10(5) RENCA cells were implanted under the renal capsule of Balb/c mice. Antisense and control ON were injected intraperitoneally daily for 14 days. Tumor weights and status of metastasis were documented after sacrifice. Furthermore, vessel density in tumor tissues was determined by CD31 immunolabeling. RESULTS: Antisense treatment of RENCA cells resulted in specific reduction of the Ki-67 protein and inhibition of cell growth. A substantial cellular uptake of labeled ON was noted in vitro and in vivo. The growth of orthotopically implantated syngeneic kidney tumors in immunocompetent mice was significantly inhibited in antisense-treated animals (p < 0.05). Furthermore, lung metastases were noted in 10% of antisense-treated animals compared to 30-40% in control groups. Immunohistochemical staining of the vessel density showed no significant difference among treatment groups. CONCLUSIONS: The results demonstrate that Ki-67-directed antisense oligonucleotides are potent inhibitors of target protein expression and proliferation of tumor cells in vitro, and of tumor growth and lung metastasis formation in murine renal cell carcinoma whereas tumor vascularization is not significantly affected.

Animals↗
PubMed2004

Gene transfer of interleukin-4 delays acute rejection of splenic allografts in rats.

Spleen transplantation is the treatment of choice for some diseases, such as hemophilia A. However, the risk and intensity of rejection after spleen transplantation is greater and more difficult to control than other types of transplant. In the present study, we demonstrated that perfusion of IL-4 expression plasmids into donor spleens pretransplantation led to overexpression of IL-4 and downregulation of IFN-gamma in situ, associated with delayed acute rejection of the allograft. Gene transfer of IL-4 may represent a potential therapeutic approach to induce tolerance to splenic allografts.

Animals↗
PubMed2004

Administration of tolerogenic dendritic cells induced by interleukin-10 prolongs rat splenic allograft survival.

The risk and intensity in splenic graft rejection are greater than in other types of transplants, because the spleen is the largest peripheral lymphoid organ and the immunosuppressive drugs administered can cause splenic dysfunction. In this study, we demonstrate that intravenous injection of interleukin-10-treated donor-type dendritic cells into recipient rats prolongs the survival of splenic allografts. Although the mechanisms are not clear, the induction of tolerance to grafted spleens seems to rely mainly on blockage of expression of the costimulatory molecule CD86, by interleukin-10, leading to enhanced apoptosis of allospecific T cells by immature and tolerogenic dendritic cells. Administration of tolerogenic cells induced by interleukin-10 may thus represent a useful approach for protection of splenic allografts. Further study is required to investigate the operative pathways and to optimize the strategy targeting dendritic cells to induce tolerance in splenic allografts.

Animals↗
PubMed2004

Recombinant adenovirus vectors activate the alternative complement pathway, leading to the binding of human complement protein C3 independent of anti-ad antibodies.

Recombinant adenoviruses are one of the most common gene transfer vectors utilized in human clinical trials, but it is also clear that systemic administration of this virus will be met by host innate and adaptive antiviral immune responses. One element of innate immunity is the complement system, a group of proteins that has evolved to rapidly recognize foreign microbes and viruses and to clear them from the circulatory system prior to their gaining entry to vulnerable host cells. Excessive complement activation can initiate or propagate a number of deleterious inflammatory responses, by release of potent cytokines and anaphylatoxins and/or by direct cellular toxicity. These reactions can progress rapidly and are factors important in serious complications, including the systemic inflammatory response syndrome and the adult respiratory distress syndrome.

Adenoviridae↗
PubMed2004

Diagnosis and management of drug-associated interstitial lung disease.

Symptoms of drug-associated interstitial lung disease (ILD) are nonspecific and can be difficult to distinguish from a number of illnesses that commonly occur in patients with non-small-cell lung cancer (NSCLC) on therapy. Identification of drug involvement and differentiation from other illnesses is problematic, although radiological manifestations and clinical tests enable many of the alternative causes of symptoms in advanced NSCLC to be excluded. In lung cancer patients, high-resolution computed tomography (HRCT) is more sensitive than a chest radiograph in evaluating the severity and progression of parenchymal lung disease. Indeed, the use of HRCT imaging has led to the recognition of many distinct patterns of lung involvement and, along with clinical signs and symptoms, helps to predict both outcome and response to treatment. This manuscript outlines the radiology of drug-associated ILD and its differential diagnosis in NSCLC. An algorithm that uses clinical tests to exclude alternative diagnoses is also described.

Algorithms↗
PubMed2004

Implantation of BM cells transfected with phVEGF165 enhances functional improvement of the infarcted heart.

BACKGROUND: Experimental data have suggested that BM-cell implantation can improve infarcted cardiac function. However, the number of implanted cells that survive is still unknown. The present study was performed to investigate whether implantation of autologous BM mononuclear cells (BM-MNCs) transfected with phVEGF165 can increase the number of surviving implanted cells, and enhance functional improvement of infarcted hearts in rabbits. METHODS: Acute myocardial infarction (AMI) in rabbits was replicated by ligating the left anterior descending coronary artery, and animals were randomly divided into the following three groups: I AMI control group (n=7); II BM-MNCs transfected with phVEGF165 implantation group (n=7); III BM-MNCs implantation group (n=7). In addition, sham-operated (n=5) rabbits were randomly selected to serve as a non-infarction control group (VI). Animals for cell implantation received intramyocardial injections of autologous BM-MNCs 14 days after AMI. Echocardiography and hemodynamic studies were performed to evaluate cardiac structure and function 28 days after implantation. The implanted sites were examined using immunofluorescence to identify the phenotypes and number of the labelled cells. Reverse transcriptase (RT)-PCR and Western blot analysis were performed to detect the expression of hVEGF165 gene and VEGF165 protein respectively. RESULTS: Failed cardiac function produced by AMI was significantly improved in Groups II and III 28 days after cell implantation. BM-MNCs transfected with phVEGF165 implantation conferred a further improvement of cardiac function, with significant changes of all assessed parameters when compared with BM-MNCs implantation alone (all P<0.05). The implanted cells demonstrated myogenic differentiation with the expression of Troponin T and organized contractile proteins. The positive staining for Factor VIII-related Ag indicated the induction of angiogenesis in the infarct area. The percentage of Brdu-positive myocyte, endothelial cells was 75+/-%, 34.1+/-4.6% in Group II, significantly higher than that in Group III(51+/-7%, 11.3+/-2.5% respectively, P<0.05). RT-PCR analysis demonstrated exclusive expression of hVEGF165 gene in Group II, and Western blot showed the expression of VEGF165 protein was significantly higher in Group II than in Group III (P<0.05). CONCLUSIONS: Implantation of BM-MNCs transfected with phVEGF165 can increase the number of implanted cells surviving, and enhance the impaired cardiac function after AMI.

Angiogenesis Inducing Agents↗
PubMed2004

Accurate Monte Carlo simulations for nozzle design, commissioning and quality assurance for a proton radiation therapy facility.

Monte Carlo dosimetry calculations are essential methods in radiation therapy. To take full advantage of this tool, the beam delivery system has to be simulated in detail and the initial beam parameters have to be known accurately. The modeling of the beam delivery system itself opens various areas where Monte Carlo calculations prove extremely helpful, such as for design and commissioning of a therapy facility as well as for quality assurance verification. The gantry treatment nozzles at the Northeast Proton Therapy Center (NPTC) at Massachusetts General Hospital (MGH) were modeled in detail using the GEANT4.5.2 Monte Carlo code. For this purpose, various novel solutions for simulating irregular shaped objects in the beam path, like contoured scatterers, patient apertures or patient compensators, were found. The four-dimensional, in time and space, simulation of moving parts, such as the modulator wheel, was implemented. Further, the appropriate physics models and cross sections for proton therapy applications were defined. We present comparisons between measured data and simulations. These show that by modeling the treatment nozzle with millimeter accuracy, it is possible to reproduce measured dose distributions with an accuracy in range and modulation width, in the case of a spread-out Bragg peak (SOBP), of better than 1 mm. The excellent agreement demonstrates that the simulations can even be used to generate beam data for commissioning treatment planning systems. The Monte Carlo nozzle model was used to study mechanical optimization in terms of scattered radiation and secondary radiation in the design of the nozzles. We present simulations on the neutron background. Further, the Monte Carlo calculations supported commissioning efforts in understanding the sensitivity of beam characteristics and how these influence the dose delivered. We present the sensitivity of dose distributions in water with respect to various beam parameters and geometrical misalignments. This allows the definition of tolerances for quality assurance and the design of quality assurance procedures.

Algorithms↗
PubMed2004

Adaptation of GEANT4 to Monte Carlo dose calculations based on CT data.

The GEANT4 Monte Carlo code provides many powerful functions for conducting particle transport simulations with great reliability and flexibility. However, as a general purpose Monte Carlo code, not all the functions were specifically designed and fully optimized for applications in radiation therapy. One of the primary issues is the computational efficiency, which is especially critical when patient CT data have to be imported into the simulation model. In this paper we summarize the relevant aspects of the GEANT4 tracking and geometry algorithms and introduce our work on using the code to conduct dose calculations based on CT data. The emphasis is focused on modifications of the GEANT4 source code to meet the requirements for fast dose calculations. The major features include a quick voxel search algorithm, fast volume optimization, and the dynamic assignment of material density. These features are ready to be used for tracking the primary types of particles employed in radiation therapy such as photons, electrons, and heavy charged particles. Recalculation of a proton therapy treatment plan generated by a commercial treatment planning program for a paranasal sinus case is presented as an example.

Algorithms↗
PubMed2004

Chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) and hepatocyte nuclear factor 4gamma (HNF-4gamma) and HNF-4alpha regulate the bovine growth hormone receptor 1A promoter through a common DNA element.

The growth hormone receptor (GHR) 1A promoter is responsible for transcription of the liver-specific GHR mRNA variant 1A in several mammalian species. We previously found that the region between nucleotide -218 and nucleotide -151 (relative to the major transcription start site) of the bovine GHR 1A promoter contained a binding site between -196 and -178 for the liver-enriched transcription factor hepatocyte nuclear factor 4alpha (HNF-4alpha) and that the same region might also interact with additional transcription factors in the liver. Using the -218/-151 region as bait to screen a bovine liver cDNA library in a yeast one-hybrid analysis, we identified chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) and HNF-4gamma, as well as HNF-4alpha, as binding proteins to the -218/-151 region. These binding proteins were also identified in a second yeast one-hybrid analysis using the -196/-178 region as bait, suggesting that COUP-TFII, HNF-4gamma and HNF-4alpha all bind to the -196/-178 region. Electrophoretic mobility shift assays confirmed the ability of these three transcription factors in bovine liver nuclear extracts to interact with the -196/-178 region. These interactions also appeared to exist in vivo, as the GHR 1A promoter containing the -196/-178 region could be recovered by immunoprecipitation of the bovine liver chromatin with antibody against COUP-TFII, HNF-4gamma or HNF-4alpha. In co-transfection analyses, each of these three transcription factors could activate GHR 1A promoter and the activation was dependent on the -196/-178 region. These results together identify COUP-TFII, HNF-4gamma and HNF-4alpha as transcription factors regulating GHR 1A promoter activity through binding to a common DNA element.

Amino Acid Sequence↗
PubMed2003

Nitric oxide-epoxygenase interactions and arachidonate-induced dilation of rat renal microvessels.

Nitric oxide (NO) is an inhibitor of hemoproteins including cytochrome P-450 enzymes. This study tested the hypothesis that NO inhibits cytochrome P-450 epoxygenase-dependent vascular responses in kidneys. In rat renal pressurized microvessels, arachidonic acid (AA, 0.03-1 microM) or bradykinin (BK, 0.1-3 microM) elicited NO- and prostanoid-independent vasodilation. Miconazole (1.5 microM) or 6-(2-propargyloxyphenyl)hexanoic acid (30 microM), both of which are inhibitors of epoxygenase enzymes, or the fixing of epoxide levels with 11,12-epoxyeicosatrienoic acid (11,12-EET; 1 and 3 microM) inhibited these responses. Apamin (1 microM), which is a large-conductance Ca2+-activated K+ (BKCa) channel inhibitor, or 18alpha-glycyrrhetinic acid (30 microM), which is an inhibitor of myoendothelial gap junctional electromechanical coupling, also inhibited these responses. NO donors spermine NONOate (1 and 3 microM) or sodium nitroprusside (0.3 and 3 microM) but not 8-bromo-cGMP (100 microM), which is an analog of cGMP (the second messenger of NO), blunted the dilation produced by AA or BK in a reversible manner without affecting that produced by hydralazine. However, the non-NO donor hydralazine did not affect the dilatory effect of AA or BK. Spermine NONOate did not affect the dilation produced by 11,12-EET, NS-1619 (a BKCa channel opener), or cromakalim (an ATP-sensitive K+ channel opener). AA and BK stimulated EET production, whereas hydralazine had no effect. On the other hand, spermine NONOate (3 microM) attenuated basal (19 +/- 7%; P < 0.05) and AA stimulation (1 microM, 29 +/- 9%; P < 0.05) of renal preglomerular vascular production of all regioisomeric EETs: 5,6-; 8,9-; 11,12-; and 14,15-EET. These results suggest that NO directly and reversibly inhibits epoxygenase-dependent dilation of rat renal microvessels without affecting the actions of epoxides on K+ channels.

8,11,14-Eicosatrienoic Acid↗
PubMed2003

Role of epoxyeicosatrienoic acids in renal functional response to inhibition of NO production in the rat.

Nitric oxide (NO) inhibits hemoproteins, including cytochrome (CYP) 2C, the gene responsible for the production of epoxyeicosatrienoic acids (EETs). EETs and NO are produced in the kidney, and both regulate renal vascular tone and Na+ transport. However, the role of EETs in NO-mediated renal function is not known. This study tested the hypothesis that NO tonically regulates the renal production of EETs, thereby impacting renal vasomotor tone and electrolyte balance. LPS (10 mg/kg i.v.) inhibited microsomal conversion of 14C-labeled arachidonic acid to EETs and reduced mean arterial blood pressure (MABP; Delta = 63 +/- 5 mmHg). Nitro-l-arginine methyl ester (l-NAME, 10 mg/kg), an inhibitor of NO synthase, increased MABP (Delta = 26 +/- 6 mmHg), reduced cortical (CBF) and medullary (MBF) blood flow (Delta = -0.86 +/- 0.15 and -0.34 +/- 0.09 V, respectively) and glomerular filtration rate (GFR; from 0.82 +/- 0.16 to 0.32 +/- 0.10 ml x g kidney-1 x min-1), and increased Na+ excretion (UNaV, from 0.16 +/- 0.04 to 0.30 +/- 0.06 micromol x g kidney-1 x min-1). 2-(2-Propynyloxy)-benzenehexanoic acid (PPOH), a suicide substrate inhibitor of EET production, did not affect the l-NAME-induced increase in MABP but attenuated the effects of l-NAME on CBF (31 +/- 7%, P < 0.05%), GFR (44 +/- 6%, P < 0.05), and UNaV (78 +/- 7%, P < 0.05). Miconazole (1.3 mg x kg-1 x h-1), a heme inhibitor of epoxygenase enzymes, produced effects similar to those of PPOH. Renal intraarterial infusion of 5,6-, 8,9-, 11,12-, and 14,15-EET (1-10 ng/min) elicited dose-dependent reductions in CBF and GFR accompanied by regioisomeric changes in MBF, UNaV, and urine flow rate. In addition, 11,12-EET dose dependently restored the PPOH blunting the effects of l-NAME on CBF, MBF, and GFR. We conclude that NO tonically regulates epoxygenase activity and that EETs are renal vaosoconstrictors in vivo and contribute, at least in part, to the renal functional responses following inhibition of NO production.

8,11,14-Eicosatrienoic Acid↗
PubMed2003

Regulatory CD8+ T cells fine-tune the myelin basic protein-reactive T cell receptor V beta repertoire during experimental autoimmune encephalomyelitis.

A significant number of self-reactive T cell clones escape thymic negative selection and are released into the periphery, where some are potentially pathogenic. The clonal expansion of self-reactive T cells is known to be limited during initial antigen encounter by apoptotic or anergic mechanisms, regulatory CD4+ T cells, and cytokines. Here we report that superimposed on these mechanisms, during the evolution of autoimmunity in experimental autoimmune encephalomyelitis (EAE), CD8+ T cells are induced, which fine-tune the peripheral self-reactive T cell receptor (TCR) repertoire. We assayed the myelin basic protein-reactive TCR repertoire in naive, EAE-recovered mice as well as EAE-recovered mice depleted of CD8+ T cells by TCRV beta surface expression, complementarity-determining region 3 length distribution, and complementarity-determining region 3 sequencing analysis. In EAE-recovered mice, certain myelin basic protein-reactive CD4+V beta 8.2+ clones are significantly decreased and this decrease is not observed if CD8+ T cells were depleted from these mice. The clones that persist in CD8+ T cell-intact mice are highly diverse in contrast to the clones expanded in CD8+ T cell-depleted mice, which are dominated by the significant outgrowth of a few clones. Importantly, the T cell clones that expand in the absence of CD8+ T cell control are enriched in potentially pathogenic self-reactive T cell clones capable of inducing EAE in vivo.

Adoptive Transfer↗
PubMed2003

Cloning and characterization of the bovine class 1 and class 2 insulin-like growth factor-I mRNAs.

Insulin-like growth factor-I (IGF-I) is an important regulator of growth, development, and metabolism, and is the primary mediator of the growth-promoting activity of growth hormone (GH) in animals. In several species, the IGF-I polypeptide is generated from IGF-I mRNA containing either exon 1 (class 1 IGF-I mRNA) or exon 2 (class 2 IGF-I mRNA) as the leader exon. The objectives of this study were to identify class 1 and class 2 IGF-I mRNAs in cattle and to compare their expression in different tissues, their response to GH, and their translational efficiency. Three class 1 IGF-I cDNAs corresponding to three different transcription start sites in exon 1 and one class 2 IGF-I cDNA were identified from adult cattle liver using 5'-rapid amplification of cDNA ends (5'-RACE). Both classes of IGF-I mRNAs were expressed in a variety of tissues, with the highest level in liver; class 1 IGF-I mRNA was more abundant than class 2 IGF-I mRNA in all tissues. Six hours after a single intramuscular injection of 500 mg of recombinant bovine GH, class 1 and class 2 IGF-I mRNAs in steer liver were increased by 29% (P=0.07) and 62% (P<0.05), respectively. The luciferase reporter mRNA fused to a class 1 IGF-I 5'-untranslated region (5'-UTR) was translated four times more efficiently in vitro than the luciferase reporter mRNA fused to a class 2 IGF-I 5'-UTR (P<0.05). These results indicate that the IGF-I gene in cattle is transcribed as class 1 and class 2 IGF-I mRNAs and that the two classes of IGF-I mRNAs may be regulated differentially at both transcriptional and translational levels.

5' Untranslated Regions↗
PubMed2003

Human catechol-O-methyltransferase down-regulation by estradiol.

Catechol-O-methyltransferase (COMT) is a crucial enzyme in dopamine and levodopa metabolism. Previously we reported that physiological concentrations of 17beta-estradiol (E2) down-regulated steady-state 1.3-kb COMT mRNA levels in MCF-7 cells. In this study, we investigated whether similar reductions occurred in a glial cell line (U138MG) and whether COMT protein and activity levels paralleled the reduction in COMT mRNA levels in MCF-7 cells. In addition, we explored the mechanism of E2 action. E2 had no effect on COMT mRNA levels in U138MG cells, but significantly reduced COMT protein and activity in MCF-7 cells (activity by 53% at 10(-7) M of E2, by 45% at 10(-8) M, and by 28% at 10(-9) M relative to non-E2-treated cells). A specific estrogen receptor antagonist (ICI 182780) blocked these estrogenic effects. Estrogen receptor in nuclear extracts of MCF-7 cells, which were pretreated with E2 (10(-9) M) for 48 h, bound to the whole proximal and distal promoter regions, as determined by electrophoretic mobility shift analysis (EMSA). We propose that E2 decreased COMT activity through down-regulation of its gene and protein expression mediated via ER interaction with response elements in the promoter region of the gene. Our findings may explain the lower of COMT activity in women compared to that in men, and, in part, the beneficial effects of E2 therapy in post-menopausal Parkinson's disease patients.

Antibody Specificity↗