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Biomedical subjects

T Li

Publications and source records attributed to T Li.

At least 379 records · Page 21Linked to original sources

Major groove recognition elements in the middle of the T7 RNA polymerase promoter.

T7 RNA polymerase recognizes a relatively small promoter extending only 17 base pairs upstream from the start site for transcription. A model for this recognition suggests that the enzyme interacts with the major groove of duplex DNA in the region centered at position -9 [Muller, D.K., et al. (1989) Biochemistry 28, 3306-3313], and recent kinetic analyses of promoters containing base analogs at positions -10 and -11 have provided support for this model [Schick, C., & Martin, C.T. (1993) Biochemistry 32, 4275-4280; Schick, C., & Martin, C.T. (1995) Biochemistry 34, 666-672]. In the current work, we extend this analysis across the proposed major groove, identifying specific base functional group contacts at positions -9 through -5. Specifically, the 6-carbonyl of guanine at positions -9 and -7, the 6-amino group of adenine at position -8, the 5-methyl group of thymine at position -6 and the 2-amino group of guanine at position -5 are identified as primary contacts. The results strongly support the model for duplex recognition in this region of the promoter and suggest that recognition continues along one face of the helix beyond the major groove and into the adjoining minor groove at position -5, where helix melting begins.

Bacteriophage T7↗

TNF-alpha potentiates oxidant and reperfusion-induced endothelial cell injury.

Pulmonary edema following reperfusion is a major clinical problem. Changes in endothelial cell shape induced by oxidant injury may account for immediate capillary leakage associated with reperfusion injury. In these experiments we examined the role of tumor necrosis factor-alpha (TNF-alpha) in acute endothelial cell injury following ischemia-reperfusion. Sprague-Dawley rats were treated with a neutralizing antisera directed against TNF-alpha prior to production of distal ischemia. These rats demonstrated a significant reduction (P < 0.05) in acute lung edema in response to 4 hr of ischemia and 30 min of reperfusion when compared to rats undergoing the same procedure without antisera treatment. An in vitro model was developed to determine if TNF-alpha had a direct effect on endothelial cell response to ischemia-reperfusion. The effects of TNF-alpha and oxidant stress on the integrity of cultured endothelial cell monolayers was measured. Rat pulmonary artery endothelial cell monolayers reacted in vitro to oxidant stress by an increase in permeability. The cells changed shape and an increase in diffusion of 125I-albumin across cell monolayers resulted when these cells were exposed to 50 microM hydrogen peroxide (H2O2) or plasma from the ischemic hind limb of a Sprague-Dawley rat (50 microliters/ml). Pretreatment of cultured cells with low levels of recombinant mouse TNF-alpha significantly affected both the cell shape change and the increase in permeability (P < 0.05). Increased permeability of cell monolayers in vitro was not due to cell lysis as determined by media lactate dehydrogenase levels. The effect appeared to be due to cellular rounding and contraction seen using video time lapse microscopy. These data suggest a direct effect of TNF-alpha on endothelial cells, whereby the cells are rendered more susceptible to oxidant injury accompanying reperfusion.

Acute Disease↗

Cationic cyclopropanation by antibody catalysis.

Reactions involving highly reactive carbocations play a central role in many important chemical processes, such as cyclization reactions. However, the potential for controlling the pathways of such reactions to obtain energetically disfavoured (but desirable) products has been hard to realize because of the difficulties inherent in controlling the conformation and chemical environment of the carbocation intermediates. Antibody catalysts, with their high specificity and binding energies, can provide the degree of conformational and chemical control necessary for directing such reactions. Here we show how antibody catalysis can guide cationic cyclization reactions selectively to form products (in high yield) that would otherwise be highly disfavoured. Most notable is the formation of a strained bicyclic compound containing a rare cyclopropane group. To explain our results, we propose a common reaction scheme in which the key step is the formation of a highly reactive protonated cyclopropane intermediate; subtle structural modifications to the substrate (the compound on which the catalytic antibody acts) lead to dramatic differences in the structure of the final product.

Antibodies, Catalytic↗

Sequence-selective carbohydrate-DNA interaction: dimeric and monomeric forms of the calicheamicin oligosaccharide interfere with transcription factor function.

The synthetic oligosaccharide moiety of the antibiotic calicheamicin and the head-to-head dimer of this oligosaccharide are known to bind to the minor groove of DNA in a sequence-selective manner preferring distinct target sequences. We tested these carbohydrates for their ability to interfere with transcription factor function. The oligosaccharides inhibit binding of transcription factors to DNA in a sequence-selective manner, probably by inducing a conformational change in DNA structure. They also interfere with transcription by polymerase II in vitro. The effective concentrations of the oligosaccharides for inhibition of transcription factor binding and for transcriptional inhibition are in the micromolar range. The dimer is a significantly more active inhibitor than is the monomer.

Aminoglycosides↗

Sensitivity-enhanced echo-planar MRI at 1.5T using a 5 x 5 mesh dome resonator.

In this work a 5 x 5 mesh dome resonator that has been optimized for functional brain imaging is presented. The resonator was reduced in length and diameter compared with previous versions to reduce sample losses, thus enhancing the signal-to-noise ratio of the acquired data. In addition, a 5 x 5 mesh design was employed, which offered improved axial homogeneity over an earlier 3 x 3 mesh version. The new resonator exhibited high sensitivity and good homogeneity over the brain volume, permitting analysis of functional activation over large areas of the cerebral cortex. In a direct comparison with a standard clinical head-imaging resonator, the high sensitivity of the 5 x 5 mesh dome resonator resulted in greater statistical confidence in functional activation.

Cerebral Cortex↗

Performance in the cross-maze and slip funnel tests of four pairs of rat lines selectively bred for divergent alcohol drinking behavior.

Alcohol naive rats from lines genetically selected for high and low alcohol drinking and founded on either Wistar (P/NP lines) and distinct heterogeneous stocks (AA/ANA and replicate HAD/LAD lines) were tested in the explorative cross-maze and the inescapable slip funnel. Rats of the low alcohol-consuming ANA, NP, LAD1 and LAD2 lines all exhibited a shorter latency before initiating exploration of the maze than did their high alcohol-consuming counterparts (66, 51, 33 and 51% of the values for AA, P, HAD1 and HAD2 lines, respectively). Significant line differences were also found with the slip funnel test (AA > ANA for time in a sprawling posture; P > NP but HAD1 < LAD1 and HAD2 < LAD2 for time escaping), but the directions of line differences were not consistently related to those in alcohol drinking.

Journal Article↗

Drug-resistant breast cancer cells frequently retain expression of a functional wild-type p53 protein.

Abnormalities in the p53 tumor suppressor gene have been shown to affect cellular processes related to cell cycle control and gene amplification. In this study we compare the status and function of wild-type p53 in MCF-7 breast cancer cells with sublines selected for resistance to chemotherapeutic agents having different mechanisms of action. Sublines that were resistant to melphalan, pyrazafurin, mitoxantrone, etoposide and PALA all retained expression of wild-type p53. Methotrexate-resistant MCF-7 cells were unusual heterozygotes that expressed a wild-type and dominant, in-frame p53 deletion mutant and the doxorubicin-resistant cells expressed only mutant p53. Analysis of the G1 checkpoint after treatment with ionizing radiation revealed that the pyrazafurin-, melphalan- and mitoxantrone-resistant cells arrested strongly in G1. The etoposide- and PALA-resistant cells had an intermediate G1 arrest phenotype and the methotrexate- and doxorubicin-resistant cells had a minimal G1 arrest phenotype. mRNA and protein analyses of downstream effector genes, including P21CIP1/Waf1, mdm2, Gadd 45 and the retinoblastoma protein, did not entirely differentiate sublines having a strong versus intermediate G1 arrest phenotype. Neither the p53 status nor the strength of the G1 arrest could be correlated with cell survival after ionizing radiation. When drug-sensitive MCF-7 cells were treated with the same chemotherapeutic agents, p53 and p21CIP1/Waf1 levels increased between 2- and 14-fold. Together these data suggest that other cellular factors likely play a role in overcoming the inhibitory effects of ionizing radiation on p53 in drug-resistant breast cancer cells.

Amides↗

Accuracy and precision of the Tomey ViVA infrared photorefractor.

PURPOSE: The Fortune Optical (Tomey ViVA) VRB-100 video refractor was tested to determine its accuracy and precision in measuring manifest refractions of human eyes with and without cycloplegia. The specific issues addressed included its accuracy in measuring spherical and cylindrical refractive errors and its precision in refracting near emmetropia. METHODS: To determine its ability to measure moderate to high (> 4.00 D) myopia, we compared the VIVA's refractions to those taken by a Canon Autorefractor R1 and a retinoscopist. A spherical lens series from -7.00 to + 7.00 D at 1.00 D intervals, or -5.00 to + 5.00 at 0.50 D intervals, was placed over a subject's eye, which was then covered by an infrared (IR) filter, refracted, and analyzed to determine the VIVA's ability to measure spheres. Subjects with refractive errors of -2.00 to + 2.00 DS (diopters sphere) and 0 to 1.00 DC (diopters cylinder) were refracted 7 to 15 times during 1 sitting to determine the VIVA's precision. The instrument's accuracy in measuring cylinders was tested by placing + 3.00 to -3.00 D cylinders (at 0.50 D intervals) over the eyes of subjects at 0 degree, 45 degrees, 60 degrees, 75 degrees, 90 degrees, 105 degrees, 120 degrees, and 135 degrees. RESULTS: The VIVA measured spheres of +/- 3.00 D with a root mean squared (rms) error of 0.5 +/- 0.1 D. Beyond this power, its accuracy progressively worsened. In some subjects, irregular intensity profiles compromised the VIVA's accuracy even with low spherical refractive errors. The VIVA was very precise in measuring spheres from + 2.00 to -2.00 D and cylinders from 0 to 1.00 D. Although the ViVA adequately measured all cylinder powers at 0 degree and 90 degrees, the accuracy of cylindrical power measurement decreased with obliquity; only cylinders < or = 1.00 D magnitude were accurately measured at 45 degrees and 135 degrees. CONCLUSIONS: We conclude that, although the ViVA offers many attractive features for vision screening, it is seriously limited by its inability to property detect and measure oblique astigmatic errors.

Humans↗

Preferential transmission of the high activity allele of COMT in schizophrenia.

Catechol-O-methyl transferase (COMT) metabolizes a variety of catecholamines such as dopamine, adrenaline and noradrenaline. It exists in common high and low activity forms. The low activity form is the result of an amino acid substitution (val-108-met) which reduces the thermostability of the enzyme [Lotta et al. (1994) Biochemistry, 34, 4202-4210]. We have genotyped this polymorphism in 178 trios consisting of Han Chinese schizophrenic subjects and their parents in order to test the hypothesis that the high activity allele is transmitted more often to affected subjects. The data were analysed using the transmission disequilibrium test (TDT), a robust method of detecting linkage in the presence of allelic associations. Of the 131 parents heterozygous at this locus, 80 transmitted the high activity allele (val-108) to affected offspring, while the remaining 51 transmitted the low activity allele (p = 0.005, one-tailed). Combining this result with that of a previous TDT study of the same polymorphism in familial schizophrenia [Kunugi et al. (1996) submitted] gives significant evidence for linkage disequilibrium (p = 0.0015). However, val-108 is frequent in the Han Chinese population, and in the present sample, 239 of the 350 non-transmitted parental alleles were val-108 (68%). It is therefore unlikely that val-108 allele of COMT has a major effect on susceptibility to schizophrenia. Our results suggest that either val-108 is a minor risk factor for schizophrenia, that the COMT gene has additional polymorphisms with greater effect on risk, or that this region of chromosome 22 contains a susceptibility gene which is in linkage disequilibrium with the COMT gene.

Adolescent↗

Synthesis of fluorinated Bis-GMA and its use with other fluorinated monomers to formulate hydrophobic composites.

Fluorinated Bis-GMA was synthesized and used with other commercially available fluorinated monomers and diluents to prepare hydrophobic composites. The composites were formulated as one-paste systems and were polymerized using blue light. Mechanical properties and water-related qualities were determined. Fluorination generally improved the hydrophobicity of the composites, but there was no clear-cut effect on mechanical properties.

Adsorption↗

Reduced liver function is the trigger for renal sodium retention following portal vein ligation in the rat.

Sodium retention along with peripheral vasodilation are features of prehepatic portal hypertension. In several models of experimental liver damage, sodium retention occurs only when hepatic function, measured by the aminopyrine breath test (ABT-k), falls below a critical threshold. The relationship between renal sodium handling, ABT-k and systemic and renal haemodynamics in partial portal vein ligated (PVL) rats was examined to test hypothesis that peripheral vasodilation was responsible for initiating sodium retention. Haemodynamic measurements were conducted early after surgery in portal hypertensive rats with and without sodium retention and in sham-operated controls. Compared with control, both PVL groups of rats had elevated portal pressure and lower peripheral vascular resistance (P < 0.05). Sodium retaining-PVL rats had both lower ABT-k (0.95 +/- 0.05 vs 1.38 +/- 0.06 x 10(-2)/min; P < 0.05) and higher sodium balance (1.38 +/- 0.09 vs 0.43 +/- 0.09 mmol/day; P < 0.05) than non-sodium retaining PVL rats. No differences in plasma renin activity or noradrenaline concentrations were observed. In a separate group of rats, hydralazine-induced pheripheral vasodilation did not induce sodium retention. These results suggest that the presence of peripheral vasodilation alone is not sufficient to trigger a sodium-retaining status. A factor, probably liver function-dependent, acting directly on renal tubules may be necessary for changes in renal sodium handling in this model.

Animals↗

Phototoxicity of some bromine-substituted rhodamine dyes: synthesis, photophysical properties and application as photosensitizers.

The synthesis of some bromine-substituted rhodamine derivatives viz., 4,5-dibromorhodamine methyl ester (dye 2) and 4,5-dibromorhodamine n-butyl ester (dye 3) are reported. These dyes were synthesized to promote a more efficient cancer cell photosensitizer for potential use in in vitro bone marrow purging in preparation for autologous bone marrow transplantation. Spectroscopic and photophysical characterization of these dyes together with rhodamine 123 (dye 1) are reported in water, methanol, ethanol and also in a microheterogeneous system, sodium dodecyl sulfate. The possible mechanism of photosensitization is characterized in terms of singlet oxygen efficiency of these dyes. Singlet oxygen quantum yields for bromine-substituted dyes are in the range of 0.3-0.5 depending on the solvent. For dye 1 no singlet oxygen production is found. The photodynamic actions of these dyes in different cell lines are tested. It was found that dye 2 and dye 3 are efficient photosensitizers and mediate eradication of K562, EM2, myeloid cell lines (CML) and the SMF-AI rhabdomyosarcoma line.

Animals↗

Isolation and characterization of a new bacterium carboxylating phenol to benzoic acid under anaerobic conditions.

A consortium of spore-forming bacteria transforming phenol to benzoic acid under anaerobic conditions was treated with antibiotics to eliminate the four Clostridium strains which were shown to be unable to accomplish this reaction in pure culture and coculture. Clostridium ghonii was inhibited by chloramphenicol (10 micrograms/ml), whereas Clostridium hastiforme (strain 3) and Clostridium glycolicum were inhibited by clindamycin (20 micrograms/ml), without the transformation of phenol being affected. Electron microscopic observations of resulting liquid subcultures revealed the presence of two different bacilli: a dominant C hastiforme strain (strain 2) (width, 1 micron) and an unidentified strain 6 (width, 0.6 micron) which was not detected on solid medium. Bacitracin (0.5 U/ml) changed the ratio of the strains in favor of strain 6. C hastiforme 2 was eliminated from this culture by dilution. The isolated strain 6 transformed phenol to benzoic acid and 4-hydroxybenzoic acid to phenol and benzoic acid in the presence of proteose peptone. Both of these activities are inducible. This strain is a gram- variable, flagellated rod with a doubling time of 10 to 11 h in the presence of phenol. It has a cellular fatty acid composition like that of C. hastiforme. However, strain 6 does not hydrolyze gelatin or produce indole. The 16S rRNA sequence of strain 6 was found to be most similar to that of some Clostridium species, with homology ranging from 80 to 86%. Tbe evolutionary relationships of strain 6 to different groups of Clostridium and Clostridium-related species revealed that it does not emerge from any of these groups. Strain 6 most likely belongs to a new species closely related to Clostridium species.

Anaerobiosis↗

The antitumor effect induced by local injections with interleukin-2 is diminished by combing with a local injection with mitomycin C.

We investigated the antitumor effect of local injections with interleukin (IL)-2 in combination with an injection with mitomycin C (MMC). In BALB/c mice inoculated intraperitoneally (i.p.) with syngeneic Meth A fibrosarcoma on day 0, the i.p. injections with IL-2 at a dose of 20000U twice daily from day 7 to day 10 significantly prolonged survival of the treated mice and such treatment augmented a concomitant immunity specific for the tumor i.p. inoculated. On the other hand, the i.p. injections with IL-2 at a dose of either 5000U or 50000U resulted in no prolonged survival of the treated mice. In addition, neither intravenous nor subcutaneous injections with IL-2 at a dose of 20000U showed the prolonged survival of the treated mice. We further investigated a modulatory effect of a local injection with MMC on the IL-2-induced antitumor effect. The in vivo antitumor effect of local injections with IL-2 was not detected when combined with a local administration of MMC, whereas a systemic administration of MMC showed no such inhibitory effect on the IL-2-induced antitumor effect. Moreover, a delayed-type hypersensitivity response against Meth A, which was augmented by the local injections with IL-2, was significantly diminished by the local administration of MMC. Collectively, our results indicate that local injections with IL-2 could elicit the antitumor effect in vivo only when given at an optimal dose and that this effect could be rather diminished by combing with a local administration of MMC.

Animals↗

[Experiment study on biological and immunological characteristics of modified DNL of oral squamous cell carcinoma with TNF-alpha gene]

Growing characteristic of gene-transduced and nontransduced DNL in medium of containing rIL-2 was compared,and growing characteristic of gene-transduced DNL in medium of non containing rIL-2 was analyzed.DNA index (DI),cell cycle and immunologic phenotypes of gene-transduced and nontransduced DNL were analyzed with FACS.The results revealed interfered with growth,DL,cell cycle and immunologic phenotypes of DNL,and gene-transduced DNL couldn't growing infinitely.

Journal Article↗