A simple improvement to the Triton lysis procedure for plasmid isolation.
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Bacteriological Techniques↗
Biomedical subjects
A Huang
Publications and source records attributed to A Huang.
NMR analysis of helix I from the 5S RNA of Escherichia coli.
The structure of helix I of the 5S rRNA from Escherichia coli has been determined using a nucleolytic digest fragment of the intact molecule. The fragment analyzed, which corresponds to bases (-1)-11 and 108-120 of intact 5S rRNA, contains a G-U pair and has unpaired bases at its termini. Its proton resonances were assigned by two-dimensional NMR methods, and both NOE distance and coupling constant information have been used to calculate structural models for it using the full relaxation matrix algorithm of the molecular dynamics program XPLOR. Helix I has A-type helical geometry, as expected. Its most striking departure from regular helical geometry occurs at its G-U, which stacks on the base pair to the 5' side of its G but not on the base pair to its 3' side. This stacking pattern maximizes interstrand guanine-guanine interactions and explains why the G-U in question fails to give imino proton NOE's to the base pair to 5' side of its G. These results are consistent with the crystal structures that have been obtained for wobble base pairs in tRNAPhe [Mizuno, H., & Sundaralingam, M. (1978) Nucleic Acids Res. 5, 4451-4461] and A-form DNA [Rabbinovich, D., Haran, T., Eisenstein, M., & Shakked, Z. (1988) J. Mol. Biol. 200, 151-161]. The conformations of the terminal residues of helix I, which corresponds to bases (-1)-11 and 108-120 of native 5S RNA, are less well-determined, and their sugar puckers are intermediate between C2' and C3'-endo, on average.
Base Composition↗
Loss of heterozygosity mapping in Wilms tumor indicates the involvement of three distinct regions and a limited role for nondisjunction or mitotic recombination.
Loss of heterozygosity (LOH) for polymorphic markers is a frequently occurring event in some tumors, reflecting the role of allele loss in the development of these tumors. We have determined LOH in 38 cases of Wilms tumor for the 2 known loci on chromosome arm 11p and for a newly detected locus on chromosome arm 16q. Only 7 of the 38 tumors studied showed reduction to homozygosity of 11p13 markers. In 4 of these tumors, reduced expression of WT1 and WIT1, genes located at 11p13 and implicated in Wilms tumorigenesis, was noted. However, this was also found in 2 of 7 tumors showing LOH exclusively of 11p15 markers and in 15 of the remaining 24 tumors in which there was no LOH for 11p markers. This suggests that events not involving mitotic recombination or chromosome nondisjunction are the most common mechanisms for mutations at the 11p Wilms tumor locus. We also noted that mitotic recombination involving 11p15 loci occurred in addition to reduced expression of the 11p13 locus genes in 2 tumors, suggesting a possible interaction between these 2 loci. In addition, LOH for 16q markers was observed in 6 tumors. In one case this was coincident with reduction of WT1 and WIT1 gene expression, and in 3 other cases it occurred in addition to 11p LOH. This indicates that an additional locus on 16q is likely to be involved in Wilms tumorigenesis.
Adolescent↗
A short questionnaire estimating risk of malnutrition.
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Aged↗
Cognitive style as a factor in accounting students' perceptions of career-choice factors.
Since prior studies of individuals' perceptions of career-choice factors have not considered the effect of cognitive styles of subjects, this study, using 68 accounting students, investigated the mediating effects of the field dependent-independent cognitive dimension on perceptions of the importance of career-choice factors. The results, in general, show that cognitive style affected individuals' perceptions of career-choice factors, suggesting that future studies of individuals' perceptions should take into account individual cognitive differences.
Accounting↗
Coordinate expression of Wilms' tumor genes correlates with Wilms' tumor phenotypes.
The cloning and molecular characterization of two putative tumor genes, WT1 and WIT1, from the chromosome 11p13 region has provided a means of evaluating their role in the generation of Wilms' tumor heterogeneity. A series of 29 tumors were analyzed for WT1 and WIT1 expression by Northern blot or RNase protection analyses, and results were compared with tumor histopathology. Tumors were scored for the percentage of mesenchymal and epithelial derived tissue components. Homotypic tumors comprised blastema, tubular epithelium, and a fibroblast-like mesenchyme. In addition to these tissue components, the group of tumors designated as heterotypic also contained ectopic cell phenotypes such as muscle and squamous epithelium. The analyses suggest that heterotypic differentiation patterns occur when WT1 and WIT1 expression is low relative to normal fetal kidney. In situ hybridization using antisense RNA probes showed that WT1 and WIT1 were concordantly expressed in normal fetal kidney and in the blastema of tumors. The ratio of WT1:WIT1 expression remained relatively constant in homotypic tumors but deviated significantly in heterotypic tumors. These results suggest that expression patterns of the WT1 and WIT1 genes can be closely correlated to Wilms' tumor histopathology.
Blotting, Northern↗
A randomized comparison of two doses of human lymphoblastoid interferon-alpha in hairy cell leukemia. Wellcome HCL Study Group.
One hundred thirty-eight patients with hairy cell leukemia were randomized to receive either a dose of 2.0 megaunits (MU)/m2 or a 10-fold lower dose of 0.2 MU/m2 of a highly purified natural alpha-interferon, administered daily for 28 days followed by a three times a week schedule. Ninety-seven of these patients had previously undergone splenectomy, but otherwise none of the patients had received prior therapy for their leukemia. The two doses were comparable in their effect on improving the neutrophil and platelet count, whereas the higher dose had a greater beneficial effect on the hemoglobin level and a greater antileukemic effect on the marrow. Acute toxicity in the form of a flu-like syndrome, neurologic side effects, neutropenia, and the need for platelet transfusions was observed less frequently in the low-dose group, as was the chronic fatigue syndrome. No neutralizing antibody activity was seen in the sera from 61 patients examined. Because of its beneficial effect on the neutrophil and platelet count and a lower degree of toxicity (ie, a superior therapeutic/toxicity ratio), the low dose is recommended as initial therapy in patients with hairy cell leukemia. This therapy may be followed by dose escalation once clinical improvement is observed.
Adult↗
EDRF-mediated shear-induced dilation opposes myogenic vasoconstriction in small rabbit arteries.
In small saline-perfused rabbit mesenteric arteries (diam 221 +/- 4 microns, means +/- SE; n = 48) in situ, the interactions of endothelium-derived relaxing factor (EDRF)-mediated flow-dependent dilation and myogenic constriction were studied. When pump flow was increased two- to fivefold (2.8 +/- 0.1-fold), input perfusion pressure rose by 133 +/- 17%. Vessel diameter first increased passively by 9 +/- 1% and then decreased to or below control values reflecting the vascular myogenic activity. This was followed by a 16 +/- 3% increase in diameter, which was flow dependent, because nonperfused vessels exposed to the same intravascular pressures did not dilate. When the perfusate viscosity was increased with dextran solutions, both the basal diameters and the flow-induced dilator responses were significantly augmented, indicating that the increase in shear stress was the stimulus. The flow-dependent dilation was abolished by inhibition of EDRF with either hemoglobin (10 microM) or NG-nitro-L-arginine (0.3 mM) and also after preincubation with neuraminidase (0.2 U/ml, 30 min), which removes part of the membrane glycocalyx. Thus, myogenic responses in small mesenteric arteries can be effectively opposed by shear-induced release of EDRF. This might be a major mechanism for maintaining adequate tissue perfusion when pressure and shear stress increase simultaneously (e.g., exercise hyperemia, autoregulation) and otherwise myogenic activity would reduce vascular conductivity.
Acetylcholine↗
Frequent identification of HIV-1 DNA in bronchoalveolar lavage cells obtained from individuals with the acquired immunodeficiency syndrome.
Tissue macrophages are recognized as a cellular target for infection with the human immunodeficiency virus type 1 (HIV-1). To characterize the nature of this cell-retrovirus interaction within the lower respiratory tract we analyzed fluid and cells obtained by bronchoalveolar lavage (BAL) of eight individuals with acquired immunodeficiency syndrome (AIDS) who were undergoing diagnostic fiberoptic bronchoscopy. Of these eight individuals, seven had active infection with Pneumocystis carinii; one had suspected cytomegalovirus pneumonitis. At the time of study two were receiving the antiretroviral drug zidovudine (azidothymidine [AZT]). HIV-1 could not be isolated from any of the eight samples of BAL fluid concentrated by ultracentrifugation through 20% sucrose. HIV-1 antigen (p24) was detected in one of eight samples of concentrated BAL fluid but could not be found in eight samples of media conditioned by overnight incubation with adherent BAL cells. Despite the infrequent detection of HIV-1 antigen it was possible to identify HIV-1 genomic sequences by the use of a DNA amplification technique, the polymerase chain reaction, in all eight BAL cell preparations. In BAL cells adherent for up to 5 days in culture this method detected retroviral DNA that hybridized to a complementary pair of primers located in the env and gag gene regions of HIV-1. These studies demonstrate the uniform presence of HIV-1 harboring cells within the airways of the lung in individuals with AIDS and active respiratory infection and may have implications for local organ defense.(ABSTRACT TRUNCATED AT 250 WORDS)
Acquired Immunodeficiency Syndrome↗
[Determination and comparison of beta-glucuronidase activity among strains of B. fragilis and E. coli].
We determined and compared the beta-glucuronidase (beta-G) activity among strains of B. fragilis and E. coli under the optimum condition. Results showed that the mean beta-G activity of strain B. fragilis ATCC25285 was 94.7u. B. fragilis ATCC25285 strain was selected as the model strain to establish the animal model of bilirubin gallstones because its beta-G activity was obviously higher than that of B. fragilis CDC14462 and E. coli 3362 (O157K88) strain.
Bacteroides fragilis↗
[Study on ursodeoxycholic acid biotransformation by three strains of clostridia].
Using the improved TLC procedure, we measured the conversion rates of ursodeoxycholic acid(UDCA) from chenodeoxycholic acid (CDCA) by 3 strains of clostridia--C. perfringens HS-10, C. butyricum DL-20 and LQ-29 isolated. The orthogonal test was used to determine the optical conditions of biotransformation for HS-10. It was found that the conversion rate of UDCA by HS-10 was over 80% during 6-48 h in RCM medium containing 0.2 mmol/L CDCA. The UDCA conversion rates were still over 70% when the CDCA concentrations were as high as 0.8-1.0 mmol/L. In addition, the WW-BCP (Waste water from bean curd production) without any supplemental nutrients was preliminarily proved to be a potential inexpensive conversion medium.
Biotransformation↗
Tissue, developmental, and tumor-specific expression of divergent transcripts in Wilms tumor.
The Wilms tumor locus on chromosome 11p13 has been mapped to a region defined by overlapping, tumor-specific deletions. Complementary DNA clones representing transcripts of 2.5 (WIT-1) and 3.5 kb (WIT-2) mapping to this region were isolated from a kidney complementary DNA library. Expression of WIT-1 and WIT-2 was restricted to kidney and spleen. RNase protection revealed divergent transcription of WIT-1 and WIT-2, originating from a DNA region of less than 600 bp. Both transcripts were present at high concentrations in fetal kidney and at much reduced amounts in 5-year-old and adult kidneys. Eleven of 12 Wilms tumors classified as histopathologically heterogeneous exhibited absent or reduced expression of WIT-2, whereas only 4 of 14 histopathologically homogeneous tumors showed reduced expression. These data demonstrate a molecular basis for the pathogenetic heterogeneity in Wilms tumorigenesis.
Base Sequence↗
Wilms tumor locus on 11p13 defined by multiple CpG island-associated transcripts.
Wilms tumor is an embryonal kidney tumor involving complex pathology and genetics. The Wilms tumor locus on chromosome 11p13 is defined by the region of overlap of constitutional and tumor-associated deletions. Chromosome walking and yeast artificial chromosome (YAC) cloning were used to clone and map 850 kilobases of DNA. Nine CpG islands, constituting a "CpG island archipelago," were identified, including three islands that were not apparent by conventional pulsed-field mapping, and thus were at least partially methylated. Three distinct transcriptional units were found closely associated with a CpG island within the boundaries of a homozygous DNA deletion in a Wilms tumor.
Chromosome Mapping↗
Epstein-Barr virus keratitis after a chemical facial peel.
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Acyclovir↗
Effects of exercise on blood flow to canine articular tissues.
We measured articular blood flow by standard microsphere techniques in normal adult dogs at rest and during treadmill exercise. All animals but one underwent beta-adrenergic blockade as part of another experimental protocol. Expressed in microliter/min/g +/- SEM, baseline flow values to articular tissues were: knee synovium 26 +/- 4, femoral condyle 130 +/- 21, tibial plateau 182 +/- 29, articular fat pad 9 +/- 2, knee ligaments 17 +/- 3, menisci 34 +/- 6, wrist synovium 19 +/- 4, distal radius 65 +/- 13, and lunate bone 59 +/- 13. Blood flow increased with exercise in all soft tissues of both the knee (stifle joint) and the wrist (radiocarpal joint). Geometric mean exercise/rest flow ratios ranged from a low of 1.44 (p less than 0.05) in the menisci of the knee to a high of 7.25 (p less than 0.001) in the synovium of the wrist. In contrast, blood flow did not rise in juxtaarticular bones and fell significantly in femoral condyles (mean flow ration 0.71, p less than 0.005). These findings indicate that articular soft tissues derive increased perfusion from the redistribution of blood flow that accompanies short-term exercise. In contrast, flow to juxtaarticular bone does not increase under these conditions.
Adrenergic beta-Antagonists↗
In vitro activity of the new semi-synthetic polypeptide cilofungin (LY121019) against Aspergillus and Candida species.
The in vitro activity of cilofungin (LY121019), a new semi-synthetic antifungal agent was evaluated. Potent activity was seen against Candida albicans and Candida tropicalis, with almost identical MIC and MFC results, whereas no activity was seen against any isolates of Candida parapsilosis or three Aspergillus spp. However, MICs were dependent on medium and test conditions chosen. It is concluded that cilofungin has good activity against some medically important yeasts in vitro, and that its in vitro activity depends on the method used.
Aspergillus↗
Definition of the limits of the Wilms tumor locus on human chromosome 11p13.
In a previous report, we described a contiguous restriction map of chromosome band 11p13 that localized the Wilms tumor locus to a small group of NotI fragments. In an effort to identify and isolate the 11p13-associated sporadic Wilms tumor locus, we developed a panel of NotI fragment-specific DNA probes. These probes were selected from genomic libraries constructed using the Chinese hamster ovary-human somatic cell hybrid carrying only human 11p. The libraries were prepared from NotI-digested DNA after size selection by pulsed-field gel electrophoresis. The selected NotI fragments had been previously targeted on the basis of deletion mapping as having a high probability of containing the Wilms tumor locus. We used these newly identified 11p13-specific probes to improve the resolution of the restriction map spanning the Wilms tumor locus. The locus has been defined by a homozygous deletion in a sporadic Wilms tumor. Using these probes, the region of homozygous deletion in this tumor and presumably all or part of the Wilms tumor gene have been confined to two small SfiI fragments spanning less than 350 kb.
Animals↗
[Application of ion chromatography to identification of anaerobic bacteria].
The fatty acids of anaerobic bacteria were detected by ion chromatography (IC). The results showed that 11 kinds of standard fatty acids could be analyzed within 15 min, and just needed 1 injection sample. The fatty acids patterns of 40 strains of anaerobic bacteria detected by IC were essentially the same as that by gas chromatography (GC) reported by VPI. In comparison with GC, IC has further advantages: pretreatment is simpler, both volatile and non-volatile fatty acids can be analyzed at the same time, and formic and lactic acid can be well analyzed.
Bacteria, Anaerobic↗