Genetic mapping of eight SH3 domain genes on seven mouse chromosomes.
Explore the source record for details and available documents.
Animals↗
Biomedical subjects
C G Lee
Publications and source records attributed to C G Lee.
The human RNA helicase A (DDX9) gene maps to the prostate cancer susceptibility locus at chromosome band 1q25 and its pseudogene (DDX9P) to 13q22, respectively.
RNA helicase A is the homolog of the Drosophila maleless protein, an essential factor involved in dosage compensation, and plays a crucial role in early development in mammals. Here, we have mapped the human RNA helicase A (DDX9) gene to the major susceptibility locus for prostate cancer at chromosome band 1q25, and its pseudogene (DDX9P) to the band 13q22 by fluorescence in situ hybridization, somatic cell hybrid analysis, and assignment of YAC clones, respectively.
Amino Acid Sequence↗
Molecular mechanism of immunoglobulin class switch recombination.
Explore the source record for details and available documents.
Animals↗
Effect of eradication of Helicobacter pylori on the benign gastric ulcer recurrence--a 24 month follow-up study.
OBJECTIVES: To evaluate the effect of eradication of Helicobacter pylori (H. pylori) on the recurrence of benign gastric ulcer (BGU) in the patients with BGU. METHODS: This study was performed for 40 H. pylori-positive BGU patients cured of BGU and H. pylori eradicated, and for 25 H. pylori-positive patients (non-eradicated group) who were not treated with H. pylori eradication regimen or H. pylori was not eradicated. Four different methods--CLOtest, microscopy of Gram stained mucosal smear, culture and histology of modified Giemsa staining--were taken for identifying colonization of H. pylori before treatment, and 4 weeks after completion of triple therapy. For the control group in which triple therapy was not tried, follow-up gastroscopy was done to confirm the healing of the ulcer. To detect BGU recurrence, the gastroscopy was performed at 6, 12, 18, and 24 months after therapy. RESULTS: In the non-eradicated group, the BGU recurrence rate was 16% within 6 months, 40% within 1 year, 56% within 18 months and 60% within 2 years. The respective recurrence rates in the 40 patients in whom the bacteria had been eradicated were 0%, 7.5%, 10% and 10% (4 patients), respectively. Among the four BGU-recurred patients in whom H. pylori had been eradicated, one patient was found to have BGU recurring with H. pylori positive again in one year, and another two patients had NSAIDs ingestion history. CONCLUSION: The eradication of H. pylori in patients with BGU reduces the recurrence of BGU. In addition, the major causes of BGU recurrence look like NSAIDs ingestion and reinfection of H. pylori.
Adult↗
RNA helicase A is essential for normal gastrulation.
RNA helicase A (RHA) is the human homologue of the Drosophila maleless protein, an essential factor for the development of male flies. Recently, it was shown that RHA cooperates with the cAMP-responsive element in mediating the cAMP-dependent transcriptional activation of a number of genes. Due to the participation of cAMP as a second messenger in a number of signaling pathways, we examined the function of RHA during mammalian embryogenesis. To examine the role(s) of RHA in mammalian development, RHA knockout mice were generated by homologous recombination. Homozygosity for the mutant RHA allele led to early embryonic lethality. Histological analysis, combined with terminal deoxynucleotidyltransferase-mediated UTP end labeling (TUNEL) reactions of RHA-null embryos, revealed marked apoptotic cell death specifically in embryonic ectodermal cells during gastrulation. RNA in situ analyses of the expression of HNF-3beta and Brachyury, two molecular markers for gastrulation, showed that RHA-null embryos at days 7.5 and 8.5 expressed both HNF-3beta and Brachyury in a pattern similar to those of pre- and early streak stages of embryos, respectively. These observations indicate that RHA is necessary for early embryonic development and suggest the requirement of RHA for the survival and differentiation of embryonic ectoderm.
Animals↗
Differences in gross target volumes on contrast vs. noncontrast CT scans utilized for conformal radiation therapy treatment planning for prostate carcinoma.
PURPOSE: To compare the gross target volumes (GTVs) (prostate and seminal vesicles) defined on noncontrast and contrast-enhanced computed tomography (CT) images used for three-dimensional conformal treatment planning (3DCRT). METHODS AND MATERIALS: From 1993 to 1996, 39 patients referred for radiation therapy for adenocarcinoma of the prostate underwent pretreatment pelvic CT scanning with and without intravenous (i.v.) contrast for treatment planning purposes. Seven patients were excluded because of incomplete data sets. The prostate and seminal vesicles were outlined by the same physician on all images of 32 patients. On 18 CT exams, the prostate and seminal vesicles were blindly outlined a second time by the same physician to evaluate intraphysician consistency. Discrepancies between the GTVs outlined with and without contrast and between the first and second outline on the same study were assessed by calculating the projected area in the anterior-to-posterior (AP) and right lateral (RLAT) beam's-eye view (BEV). To assess the magnitude, frequency, and direction of discrepancies between the two GTVs, the extension of the GTVs in six directions (right, left, anterior, posterior, cephalad, and caudal) was determined. RESULTS: The GTV outlined with contrast was larger in all directions, except caudal, in the majority of patients. The change in the GTV with contrast was significant in the cephalad (p=0.0003) and right (p=0.0007) directions, but not in the other directions. Although the increase with contrast in any direction was usually small (average < or =5 mm), these changes resulted in a significant increase in GTV area in both the AP and RLAT BEV (9.0%, p=0.0017 and 8.2%, p=0.023, respectively). The intraphysician variability in outlining the prostate/ seminal vesicles was minimal. CONCLUSIONS: The addition of i.v. contrast does appear to make a significant difference in how the prostate and seminal vesicles are outlined by an experienced observer. The increase in area of the target, found when contrast is used, should be taken into consideration when designing the treatment fields for patients with carcinoma of the prostate.
Contrast Media↗
HIV-1 protease inhibitors are substrates for the MDR1 multidrug transporter.
The FDA approved HIV-1 protease inhibitors, ritonavir, saquinavir, and indinavir, are very effective in inhibiting HIV-1 replication, but their long-term efficacy is unknown. Since in vivo efficacy depends on access of these drugs to intracellular sites where HIV-1 replicates, we determined whether these protease inhibitors are recognized by the MDR1 multidrug transporter (P-glycoprotein, or P-gp), thereby reducing their intracellular accumulation. In vitro studies in isolated membrane preparations from insect cells infected with MDR1-expressing recombinant baculovirus showed that these inhibitors significantly stimulated P-gp-specific ATPase activity and that this stimulation was inhibited by SDZ PSC 833, a potent inhibitor of P-gp. Furthermore, photoaffinity labeling of P-gp with the substrate analogue [125I]iodoarylazidoprazosin (IAAP) was inhibited by all three inhibitors. Cell-based approaches to evaluate the ability of these protease inhibitors to compete for transport of known P-gp substrates showed that all three HIV-1 protease inhibitors were capable of inhibiting the transport of some of the known P-gp substrates but their effects were generally weaker than other documented P-gp modulators such as verapamil or cyclosporin A. Inhibition of HIV-1 replication by all three protease inhibitors was reduced but could be restored by MDR1 inhibitors in cells expressing MDR1. These results indicate that the HIV-1 protease inhibitors are substrates of the human multidrug transporter, suggesting that cells in patients that express the MDR1 transporter will be relatively resistant to the anti-viral effects of the HIV-1 protease inhibitors, and that absorption, excretion, and distribution of these inhibitors in the body may be affected by the multidrug transporter.
3T3 Cells↗
Frequent but biased class switch recombination in the S mu flanking regions.
Immunoglobulin (Ig) heavy chain class switch recombination occurs mainly by joining two switch (S) regions, segments of tandemly repeated DNA sequences that lie upstream of heavy chain constant region genes. The products of this recombination event are a chromosomal DNA joint and a 'looped-out' circular DNA joint. Although a previous study showed that 40% of chromosomal joints in the mu gene switch region (S mu) are found in the flanking regions of S mu, which do not contain typical S mu region repeats [1], other studies revealed that almost all recombination sites on looped-out circular DNA are found within S regions [2-4]. To resolve this discrepancy, we have isolated and sequenced 164 DNA fragments containing recombination joints from both chromosomal and looped-out DNA of a single cell line, the murine B lymphoma line CH12F3, which switches from IgM to IgA production with a high frequency upon cytokine stimulation [5]. The recombination sites were distributed almost evenly in the S mu region and its flanking regions, suggesting that the final joining of DNA ends may not necessarily take place in S regions. In contrast, there were few joining sites in the exon located 5' of the switch region (the I mu exon), suggesting that the 3' end of the I mu exon might be the upstream border of the recombination joint.
Animals↗
Molecular analysis of the cDNA and genomic DNA encoding mouse RNA helicase A.
RNA helicase A is an enzyme that possesses both RNA and DNA helicase activities. In this report, we describe the isolation of a mouse cDNA encoding RNA helicase A. The deduced amino acid sequence derived from mouse RNA helicase A cDNA exhibits 87 and 47% identity to its human and Drosophila homologs, respectively. Using Southern blot analysis employing a mouse backcross panel, we have assigned the mouse RNA helicase A gene to chromosome 1, mapping near the D1Bir20 locus at MGD position 67. Northern blot and primer extension analyses indicate that, although its level is variable, RNA helicase A appears to be expressed from a single transcription start site in all tissues tested. Sequence analysis of the upstream genomic DNA revealed that the promoter region lacks a TATA box and contains two high-affinity sites for Sp1, one ISRE, a binding site for interferon regulatory factor, and three AP2-binding sites. These findings suggest that the transcriptional regulation of the RNA helicase A gene is complex.
Amino Acid Sequence↗
HIV-1 protease inhibitors and the MDR1 multidrug transporter.
Explore the source record for details and available documents.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Retroviral transfer of human MDR1 gene into human T lymphocytes.
Explore the source record for details and available documents.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
Target specificity of immunoglobulin class switch recombination is not determined by nucleotide sequences of S regions.
We describe a model system for class switch recombination (CSR) using CH12F3-2 cells transfected with a DNA construct containing two S sequences transcribed by different promoters and separated by a viral thymidine kinase (TK) gene. Recombination observed using this system shares key properties with physiological CSR: deletion of DNA between two S regions, requirement for cytokine stimulation, and nonhomologous and no consensus breakpoint sequences. Studies on transfectants with variants of this construct led us to the following conclusions: (1) two S sequences are required for CSR; (2) isotype specificity of recombination is not determined by nucleotide sequences of S regions; (3) S sequences are not strand-specific; and (4) induction of recombination activity requires cytokine stimulation.
Animals↗
Limb reduction defects in endothelial nitric oxide synthase-deficient mice.
Nitric oxide synthases are a family of enzymes capable of converting L-arginine to L-citrulline with the subsequent release of nitric oxide (NO). NO has been shown to have multiple biologic effects depending on the isoform responsible for its production and its tissue of origin. Murine endothelial nitric oxide synthase (eNOS) is encoded by Nos3, located on mouse chromosome 5. NO produced from this isoform causes vascular smooth muscle relaxation. Other investigators have shown that the administration of nonspecific inhibitors of nitric oxide synthases to pregnant rats induces limb reduction defects. However, mice deficient in Nos3 have not previously been noted to show such abnormalities. To explore the importance of eNOS during development, we produced mice deficient in eNOS using embryonic stem cell technology. Limb reduction defects were seen in approximately 10% of the null animals. We also observed increased neonatal loss of homozygous deficient pups. One functional copy of Nos3 eliminates the risk of limb defects observed in our mouse strain. These findings have implications for understanding genetic predisposition to sporadic limb reduction defects in humans.
Acetylcholine↗
Detection of MYC gene amplification in malignant lymphomas.
The role of oncogene or tumor supressor gene in Epstein-Barr virus (EBV) associated malignant lymphomas (MLs) is poorly understood. We examined 36 MLs (21 EBV positive and 15 EBV negative) and 6 reactive hyperplasias for the presence of myc gene amplification. Polymerase chain reaction (PCR) technology was used to examine the state of amplification of the proto-oncogene c-myc in formalin-fixed paraffin-embedded tissues. Variable degrees of myc gene amplification were detected in reactive hyperplasias and MLs. However, significant increase of c-myc copy numbers above 3 times were only found in 12 out of 31 non-Hodgkin's MLs (38.7%), in which 6 cases were EBV positive and 6 cases were EBV negative. In conclusion, myc gene amplification appears to play a part in MLs but no correlation was found between EBV infection and myc gene amplification.
Castleman Disease↗
Detection of Cryptosporidium oocysts in canine fecal samples by immunofluorescence assay.
Fecal samples were collected from 257 dogs in four areas in Korea during the period of January 1996 to November 1997 and examined by immunofluorescence assay for Cryptosporidium oocysts using a commercial diagnostic kit (Meridian Diagnostics, Cincinnati, Ohio). Of the 257 samples, 25 (9.7%) were positive for Cryptosporidium. Differences were noted in the prevalence of canine cryptosporidiosis in both areas and dog types. The results provide a further evidence of environmental contamination and widespread distribution of the parasite in Korea.
Animals↗
Combination of bupivacaine scalp circuit infiltration with general anesthesia to control the hemodynamic response in craniotomy patients.
BACKGROUND: Sudden and overwhelming increases in blood pressure (BP) and heart rate (HR) during incision of the scalp may give rise to morbidity or mortality in patients with intracranial pathology undergoing neurosurgery. A modification of the method proposed by Labat to abate this circumstantiality was applied in a group of patients receiving craniotomy. The modified method was to combine scalp circuit infiltration of local anesthetic with general anesthesia to control the hemodynamic response to craniotomy. METHODS: Twenty-six patients scheduled to undergo craniotomy were randomly divided into two groups. Patients whose conditions or their current medication that might affect the stability of hemodynamics were excluded. In group A patients (N = 16) 25-30 ml of 0.25% bupivacaine was used for scalp circuit infiltration on the operation side, while in those of group B (N = 10) the same volume of 0.9% normal saline was used. After induction, anesthesia was maintained with 0.6% to 1.2% end-tidal isoflurane (ET-Iso) and 50% N2O in oxygen (N2O:O2 = 2 l/min:2 l/min). The end-tidal CO2 was kept within the range of 25-30 mmHg. BP and HR were recorded every five min before incision and then every two min after incision until one hour after induction. ET-Iso was also recorded every two min throughout a period of sixty min. If the BP and HR increased above 20% of the baseline (10 min before incision), thiopental 2.5 mg/kg and fentanyl 2 micrograms/kg were administered. If hypertension became sustained, the isoflurane concentration was adjusted until an acceptable level was obtained. RESULTS: The mean BP during the surgery was 92 +/- 1 mmHg in group A and 92 +/- 7 mmHg in group B. The difference in BP between incision to 6 min after incision was statistically significant (P < 0.05). The mean HR during surgery was 101 +/- 5 beats/min in group B and 91 +/- 2 beats/min in group A, the difference of which was not statistically significant. All of the patients in group B required a deepened anesthesia to keep the BP and HR within the normal range, but no patient in group A had such need. The average concentration of ET-Iso during the 60 min period was 0.95 +/- 0.12% in group B and 0.41 +/- 0.01% in group A, respectively. The difference was statistically significant (P < 0.05). CONCLUSIONS: Our results showed that scalp circuit infiltration with 0.25% bupivacaine significantly improved the cardiovascular stability and reduced the requirement of isoflurane during craniotomy. The routine use of bupivacaine scalp circuit infiltration in patients undergoing craniotomy should be considered.
Adult↗
RNA helicase A mediates association of CBP with RNA polymerase II.
The coactivator CBP has been proposed to stimulate the expression of certain signal-dependent genes via its association with RNA polymerase II complexes. Here we show that complex formation between CBP and RNA polymerase II requires RNA helicase A (RHA), a nuclear DNA/RNA helicase that is related to the Drosophila male dosage compensation factor mle. In transient transfection assays, RHA was found to cooperate with CBP in mediating target gene activation via the CAMP responsive factor CREB. As a mutation in RHA that compromised its helicase activity correspondingly reduced CREB-dependent transcription, we propose that RHA may induce local changes in chromatin structure that promote engagement of the transcriptional apparatus on signal responsive promoters.
Animals↗
The NTPase/helicase activities of Drosophila maleless, an essential factor in dosage compensation.
Drosophila maleless (mle) is required for X chromosome dosage compensation and is essential for male viability. Maleless protein (MLE) is highly homologous to human RNA helicase A and the bovine counterpart of RNA helicase A, nuclear helicase II. In this report, we demonstrate that MLE protein, overexpressed and purified from Sf9 cells infected with recombinant baculovirus, possesses RNA/DNA helicase, adenosine triphosphatase (ATPase) and single-stranded (ss) RNA/ssDNA binding activities, properties identical to RNA helicase A. Using site-directed mutagenesis, we created a mutant of MLE (mle-GET) that contains a glutamic acid in place of lysine in the conserved ATP binding site A. In vitro biochemical analysis showed that this mutation abolished both NTPase and helicase activities of MLE but affected the ability of MLE to bind to ssRNA, ssDNA and guanosine triphosphate (GTP) less severely. In vivo, mle-GET protein could still localize to the male X chromosome coincidentally with the male-specific lethal-1 protein, MSL-1, but failed to complement mle1 mutant males. These results indicate that the NTPase/helicase activities are essential functions of MLE for dosage compensation, perhaps utilized for chromatin remodeling of X-linked genes.
Acid Anhydride Hydrolases↗