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Biomedical subjects

Y Chu

Publications and source records attributed to Y Chu.

At least 19 recordsLinked to original sources
PubMed1999

Assay of 6-mercaptopurine and its metabolites in patient plasma by high-performance liquid chromatography with diode-array detection.

A reversed-phase high-performance liquid chromatography (HPLC) method was developed to determine 6-mercaptopurine (MP) and seven of its metabolites (6-thioguanine, 6-thioxanthine, 6-mercaptopurine riboside, 6-thioguanosine, 6-thioxanthine riboside, 6-methylmercaptopurine and 6-methylmercaptopurine riboside) simultaneously in human plasma. A volume of 100 microl of plasma was used. Protein was removed from the sample by a simple and easy ultrafiltration step and ultrafiltrate was directly injected onto the HPLC system. Analytes were detected and confirmed with a diode-array detector before quantitation at 295 and 330 nm. The limit of detection for the analytes ranged from 20 to 50 nM. For the majority of patients receiving a 1 g/m2 MP intravenous infusion, MP and all metabolites except 6-thioguanine and 6-methylmercaptopurine riboside were present. This method serves as useful tool to characterize pharmacokinetics and pharmacodynamics of MP in oncology patients, and the small volume of plasma lends itself to pediatric studies.

Antimetabolites, Antineoplastic↗
PubMed1999

Examining the immune response in sentinel lymph nodes of mice and men.

Recently, it was recognized that an immune response develops along one of two major pathways. One leads to a destructive immune response (type 1), while the alternative leads to a nondestructive immune response (type 2). Our studies in animal models suggest that therapeutic vaccines induce a tumor-specific type 1 immune response while ineffective vaccines induce a type 2 response. These results have led us to examine the immune response in sentinel lymph nodes draining tumor vaccines of patients entered onto clinical trials for melanoma, breast and renal cell cancer.

Animals↗
PubMed1999

Mechanisms of photoreceptor death and survival in mammalian retina.

The mammalian retina, like the rest of the central nervous system, is highly stable and can maintain its structure and function for the full life of the individual, in humans for many decades. Photoreceptor dystrophies are instances of retinal instability. Many are precipitated by genetic mutations and scores of photoreceptor-lethal mutations have now been identified at the codon level. This review explores the factors which make the photoreceptor more vulnerable to small mutations of its proteins than any other cell of the body, and more vulnerable to environmental factors than any other retinal neurone. These factors include the highly specialised structure and function of the photoreceptors, their high appetite for energy, their self-protective mechanisms and the architecture of their energy supply from the choroidal circulation. Particularly important are the properties of the choroidal circulation, especially its fast flow of near-arterial blood and its inability to autoregulate. Mechanisms which make the retina stable and unstable are then reviewed in three different models of retinal degeneration, retinal detachment, photoreceptor dystrophy and light damage. A two stage model of the genesis of photoreceptor dystrophies is proposed, comprising an initial "depletion" stage caused by genetic or environmental insult and a second "late" stage during which oxygen toxicity damages and eventually destroys any photoreceptors which survive the initial depletion. It is a feature of the model that the second "late" stage of retinal dystrophies is driven by oxygen toxicity. The implications of these ideas for therapy of retinal dystrophies are discussed.

Age Factors↗
PubMed1999

Localization of IgG in the normal and dystrophic rat retina after laser lesions.

PURPOSE: To test the hypothesis that access to extravasated plasma protein IgG may influence photoreceptor survival following laser photocoagulation and to determine whether this correlates with the retinal glial reaction. METHODS: A total of 45 rats (18 Royal College of Surgeons (RCS) dystrophic and 18 RCS-rdy+ congenic control) were used for this experiment. Nine non-lasered littermates of same age were used as controls. The superior retinas of postnatal day 23 rats were irradiated with a grid pattern of 40 argon green laser lesions of 50 microm in diameter and two powers (150 and 300 mW) for 0.2 s. At various times after laser lesions (up to 14 days), animals were perfused, the retinas snap frozen and sectioned on a cryostat. A one-step immunohistochemical technique was used by incubating with rabbit anti-rat IgG conjugated directly to horseradish peroxidase. Adjacent sections were processed using an antibody to glial fibrillary acidic protein (GFAP) by the standard avidin-biotin complex method. RESULTS: The labelling pattern for extravasated IgG after laser lesion was very similar in both RCS and RCS-rdy+ rat retinas. At 6, 12 and 24 h after lesions, IgG immunoreactivity (IR) was very intense in the lesion core and flanks. The outer plexiform layer (OPL) and photoreceptor inner segments provided a ready pathway for lateral spread of IgG. However, in the outer nuclear layer (ONL), IgG localization was much more restricted. Despite very intense IgG IR in the ONL of the coagulated lesion core, there was always a very sharply delineated boundary where the label abruptly halted. The GFAP labelling in both RCS dystrophic and RCS-rdy+ congenic control rat retinas showed that this boundary was between normal and necrotic cells because there was a core where GFAP was not produced by Müller cells. By 2 days after lesions, the coagulated cells in the lesion core were being removed by phagocytic cells that were IgG IR. Labelled phagocytic cells were also found among the inner and outer segment region on the lesion flanks. There was still IgG IR in the lesion, but the label was faint. No IgG IR was found in the retina at 3, 4, 7 and 14 days after lesions. Absorption control with pure rat IgG showed the label to be specific. CONCLUSIONS: The extravasated IgG was derived from the choroidal circulation because at no stage was IgG localized around the retinal vasculature. The IgG labelling was surprisingly widespread and, therefore, did not correlate with photoreceptor sparing, although it preceded the widespread Müller cell expression of GFAP and may, therefore, trigger glial reaction.

Animals↗
PubMed1998

Constitutive telomerase activity in cells with tissue-renewing potential from estrogen-regulated rat tissues.

To investigate the role of telomerase in estrogen-regulated rodent tissues, we assayed the activity levels of this enzyme and measured cell proliferation and indicators of cellularity in vagina, mammary gland, and uterus from virgin, pregnant, ovariectomized, and ovariectomized estradiol-treated rats. No association was observed between telomerase activity and increased cell proliferation. Telomerase activity was significantly higher (P=0.003) in vagina obtained from ovariectomized rats (very low proliferation) than in vagina from ovariectomized and estradiol-treated rats (high proliferation, high differentiation). The high telomerase levels observed in vagina from ovariectomized rats indicates that the same epithelial compartment (i.e., basal layer) that has the potential to reconstitute the epithelium also contains the cells that express telomerase. The lower telomerase activity in the keratinized (differentiated) vagina was probably due to dilution of the number of telomerase-producing cells by the terminally differentiated non-telomerase-producing cells. Similar results were observed in uterus from ovariectomized versus ovariectomized and estradiol-treated rats. Telomerase activity was highest in uterus from pregnant rats. Telomerase levels in samples from total mammary gland fat pads varied considerably between groups and appeared to be representative of the amount of epithelium present in the sample. Interestingly, when mammary gland samples from the same animals were obtained from pure epithelial organoid preparations, no differences in telomerase activity could be distinguished between animals or groups. Overall these data suggest that telomerase activity, particularly in rat vagina and uterus, appears to be associated with a cell subpopulation showing proliferative and tissue reconstitution potential and not directly associated with proliferation status per se.

Animals↗
PubMed1998

Treatment of severe aplastic anemia with an immunosuppressive agent plus recombinant human granulocyte-macrophage colony-stimulating factor and erythropoietin.

To evaluate the therapeutic potential of hematopoietic growth factors (HGFs) during immunosuppressive treatment (IST) of severe aplastic anemia (SAA), 38 patients with newly diagnosed SAA received IST alone (group I), or IST plus recombinant human erythropoietin and granulocyte-macrophage colony-stimulating factor (rhEPO + rhGM-CSF) (group II). Eleven patients in each group received antilymphocyte globulin (ALG) for IST, and eight patients in each group received cyclosporine (CSA). Complete remission rates at one year were 26% and 74% for group I and group II patients, respectively. The ALG-treated subgroup showed the greatest differences between treatments. Compared with patients receiving ALG alone, patients treated with ALG plus HGFs had significantly better one-year survival (100% vs. 54.5%, P < 0.05), complete remission rates (91% vs. 36%, P < 0.05), more rapid and complete hematologic recovery, greater reductions in transfusion requirements, and lower infection rates. The data suggest a potential role for rhEPO + rhGM-CSF therapy in SAA patients receiving IST.

Adolescent↗
PubMed1998

Direct gene transfer to the respiratory tract of mice with pure plasmid and lipid-formulated DNA.

Direct gene transfer into the respiratory system could be carried out for either therapeutic or immunization purposes. Here we demonstrate that cells in the lung can take up and express plasmid DNA encoding a luciferase reporter gene whether it is administered in naked form or formulated with cationic liposomes. Depending on the lipid used, the transfection efficiency with liposome-formulated DNA may be higher, the same as, or less than that with pure plasmid DNA. Tetramethyltetraalkylspermine analogs with alkyl groups of 16 or 18 carbons and DMRIE/cholesterol formulations proved particularly effective. Similar results for reporter gene expression in the lung were obtained whether the DNA (naked or lipid formulated) was administered by indirect, noninvasive intranasal delivery (inhaled or instilled) or by invasive, direct intratracheal delivery (injected or via a cannula). Reporter gene expression peaks around 4 days, then falls off dramatically by 9 days. The dose-response is linear, at least up to 100 microg plasmid DNA, suggesting better transfection efficiencies might be realized if there was not a volume limitation. For a given dose of DNA, the best results are obtained when the DNA is mixed with the minimum amount of lipid that can complex it completely. These results are discussed in the context of direct gene transfer for either gene therapy or delivery of a mucosal DNA vaccine.

Administration, Inhalation↗
PubMed1998

Study of the relatedness of isolates of Shigella flexneri and Shigella sonnei obtained in 1986 and 1987 and in 1994 and 1995 from Hong Kong.

We used pulsed-field gel electrophoresis (PFGE) to study the genetic relatedness of 235 isolates of Shigella flexneri and Shigella sonnei collected in Hong Kong (97 isolates from 1986 and 1987 and 138 isolates from 1994 and 1995). Altogether, 13 gels were run with bacteriophage lambda ladder DNA (Pharmacia) as an external reference in every sixth lane, standardized reagents and methods, and isolates randomized for species and years. For quantitative illustration of the relationships within a large body of isolates, computer-generated dendrograms were used to determine the number of isolates in pulsotypes at Dice coefficients of similarity of 75% (PT75) and 50% (PT50). For S. flexneri, there was a significant difference in the distribution of isolates collected during the two periods in both PT75 and PT50, with 68% of isolates collected in 1994 and 1995 sharing a coefficient of similarity of >/=68%. For S. sonnei, a significant difference was observed in PT50 only. We also used Upholt's formula for an approximation of the fraction of nucleotide difference between isolates and Molecular Evolutionary Genetics Analysis to determine relative genetic distances. For both species, the relative genetic distances between isolates of the earlier collection period were significantly greater (P < 0.0001), i. e., they were further apart and therefore more diverse than those of the later period. We conclude that it is possible for a typical clinical laboratory to analyze a large amount of PFGE information on Shigella isolates obtained under controlled conditions. Such data analysis should enhance surveillance capabilities and give indications of further work to be done on various aspects of bacterial pathogenicity of the species.

Bacteriophage lambda↗
PubMed1998

Vascular effects of LPS in mice deficient in expression of the gene for inducible nitric oxide synthase.

The inducible isoform of nitric oxide synthase (iNOS) is expressed after systemic administration of lipopolysaccharide (LPS). The importance of expression of iNOS in blood vessels is poorly defined. Because nitric oxide from iNOS may alter vasomotor function, we examined effects of LPS on vasomotor function in carotid arteries from iNOS-deficient mice. We studied contraction of the carotid artery from wild-type and iNOS-deficient mice in vitro 12 h after injection of LPS (20 mg/kg ip). Contractile responses to PGF2alpha (3-30 microM) and thromboxane A2 analog (U-46619; 3-100 nM) were evaluated using vascular rings from mice treated with vehicle or LPS. Maximum force of contraction generated by rings in response to PGF2alpha was 0.39 +/- 0.02 and 0.25 +/- 0.01 (SE) g (n = 14) in vehicle and LPS-treated wild-type mice, respectively (P < 0.001 vs. vehicle). Thus LPS reduced constrictor responses in wild-type mice. Thiocitrulline and aminoguanidine (inhibitors of iNOS) improved contractile responses from LPS-treated wild-type vessels. Indomethacin also improved constrictor responses in arteries from wild-type mice injected with LPS. In contrast, contraction of the carotid arteries in response to PGF2alpha and U-46619 was not impaired in LPS-treated iNOS-deficient mice, and contraction was not altered by inhibitors of iNOS. Expression of iNOS mRNA was confirmed using RT-PCR in carotid arteries from wild-type mice after injection of LPS but not vehicle. PCR products for iNOS were not observed in iNOS-deficient mice. These findings provide the first direct evidence that iNOS mediates impairment of vascular contraction after treatment with LPS.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1998

Cationic polymer and lipids enhance adenovirus-mediated gene transfer to rabbit carotid artery.

BACKGROUND AND PURPOSE: Improvement of efficiency of gene transfer to endothelium could be useful for several applications. We tested the hypothesis that cationic nonviral molecules augment adenovirus-mediated gene transfer to blood vessels, perhaps by alteration of the surface charge of adenovirus and facilitation of binding to endothelium. METHODS: Carotid arteries from rabbits were incubated in vitro for 0.5 to 2 hours with an adenoviral vector alone or noncovalent complexes of adenovirus with poly-L-lysine (a cationic polymer) or lipofectin (a cationic lipid). Binding of adenovirus to the vessels was evaluated immediately after incubation with virus, and assay of transgene (ss-galactosidase) activity and histochemistry were performed 24 hours after gene transfer. To determine whether cationic molecules can be used to augment alteration of vascular function by adenovirus-mediated gene transfer, we also examined effects on gene transfer of endothelial nitric oxide synthase. RESULTS: Assay of ss-galactosidase activity indicated that both cationic molecules increased transgene expression in vessels by approximately 5- to 6-fold. In contrast, when endothelium was removed from the vessels after gene transfer, poly-L-lysine and lipofectin did not significantly increase transgene activity. Histochemistry for ss-galactosidase also suggested that the adenovirus-cationic molecule complexes augmented transgene expression mainly in the endothelium. In addition, we found that complexing adenovirus with cationic molecules increased binding of adenovirus to the vessels. After gene transfer with recombinant adenovirus containing endothelial nitric oxide synthase, calcium ionophore (A23187) produced greater relaxation of vessels treated with adenovirus complexed with poly-L-lysine or lipofectin than those treated with adenovirus alone. CONCLUSIONS: Cationic molecules improve the efficiency of adenovirus-mediated gene transfer to blood vessels.

Adenoviridae↗
PubMed1998

Immunophenotype of myeloid cells in myelodysplastic syndromes and its clinical implications.

OBJECTIVE: To explore the immunophenotype of myeloid cells in myelodysplastic syndyomes (MDS) and its clinical implications. METHODS: A panel of monoclonal antibody was used to detect CD13+, CD33+, CD15+ and CD14+ antigens on the membrane surfaces of myeloid cells in the bone marrow from 51 MDS, 21 aplastic anemia (AA), 21 paroxysmal nocturnal hemoglobinuria (PNH) patients. 10 acute myeloblastic leukemia (AML) patients and 15 normal subjects by immunoenzymatic assay. The morphology and chromosome karyotype of bone marrow cells of MDS patients were also examined. RESULTS: CD14+, CD13+ and CD33+ cells in the bone marrow were more in MDS patients than in normal controls, AA patients and PNH patients. CD15+ cells in the bone marrow were less in MDS patients than in normal controls. CONCLUSIONS: The percentages of CD14+, CD13+ and CD33+ positive cells in the bone marrow of MDS patients were related to the percentage of myeloblasts, the chromosomal aberrations and the response to treatment. It indicated that there is immunophenotypic misexpression of myeloid cells in MDS patients. Immunophenotype analysis of myeloid cells might be useful for the diagnosis and treatment of MDS patients.

Adolescent↗
PubMed1998

Endothelium-dependent relaxation of canine pulmonary artery endothelium after prolonged preservation.

OBJECTIVE: Experiments were designed to investigate the effect of Euro-Collins (EC) solution and University of Wisconsin (UW) solution on function of pulmonary arterial endothelium. METHODS: Third order canine pulmonary artery segments were preserved in cold (4 degrees C) UW (group 1, n = 8) or EC (group 2, n = 9) solutions for 16 hours. The preserved (group 1 and 2) and control (group 3, n = 7) pulmonary arterial segments with and without endothelium were studied in vitro in organ chambers to measure isometric tension. RESULTS: The endothelium-dependent relaxation to acetylcholine and adenosine diphosphate of group 1 and 3 were significantly better than those of group 2. CONCLUSIONS: We concluded that endothelium-dependent relaxation of canine pulmonary arterial endothelium to receptor-dependent acetylcholine and adenosine diphosphate were impaired after preservation with Euro-Collins solution. However, endothelium-dependent relaxation of pulmonary segments were well maintained after preservation with University of Wisconsin solution.

Animals↗
PubMed1998

Immunocytochemical localization of basic fibroblast growth factor and glial fibrillary acidic protein after laser photocoagulation in the Royal College of Surgeons rat.

PURPOSE: Argon laser photocoagulation slows photoreceptor degeneration in the Royal College of Surgeons (RCS) rat, as does intravitreal injection of basic fibroblast growth factor (bFGF). We hypothesize that up-regulation of retinal bFGF is a consequence of laser lesioning in RCS rats. Therefore, we examined the localization of bFGF after laser and correlated this with Mailer cell glial fibrillary acidic protein (GFAP) expression, which is known to increase after injury. METHODS: A total of 34 RCS rats at postnatal day 23 were anaesthetized (ketamine 40 mg/kg) and their retinas were irradiated with a grid pattern of 40 non-overlapping argon green lesions with a power of 120 mW for 0.2 s using a 50 microm spot size. At 0, 6, 12, 24 and 48 h and 7, 14 and 21 days post-lesion, rats were anaesthetized and their eyes were enucleated and cryostat sectioned and the sections were processed using either an antibody to bFGF or GFAP using the standard avidin-biotinylated peroxidase complex method. Five age-matched RCS rats without laser lesions served as controls. RESULTS: Basic fibroblast growth factor immunoreactivity (IR) was normally located within cells in the ganglion cell layer inner nuclear layer and in retinal pigment epithelium cells and in the extracellular matrix/cell membranes of the outer nuclear layer (ONL). In lasered retinas, there was elevated bFGF-IR in the coagulated outer segments for the first 24 h. Retinal blood vessels/Müller cells/astrocytes were moderately labelled in and near each lesion immediately after lesion and became more intense after 48 h and persisted for at least 21 days. There was an elevation of bFGF-IR in the ONL on the lesion flanks at 14 days. Muller cell GFAP-IR was first detected at 6 h post-lesion and spread for a considerable distance beyond the lesion site. At 7 and 14 days, Müller cells at the lesion site had sprouted, while those on the flanks were still GFAP-IR. CONCLUSIONS: Following laser lesion there was an increase in bFGF at the lesion core only for the first 24 h. However, elevated levels of bFGF were observed in the ONL at 14 days, which extended into the lesion flanks for a similar distance to that over which increased photoreceptor survival is found. These results provide support for the hypothesis that laser lesions induce bFGF and this may be the mechanism whereby photoreceptors are spared. Müller cell activation is consistent with growth factor stimulation, but was more widespread than the bFGF changes in ONL. However, blood vessel labelling was similarly widespread and so the responses may be linked between Müller cell GFAP reaction and blood vessel bFGF localization after laser lesions.

Animals↗
PubMed1997

Protective effect of encapsulated cells producing neurotrophic factor CNTF in a monkey model of Huntington's disease.

Huntington's disease is a genetic disorder that results from degeneration of striatal neurons, particularly those containing GABA (gamma-aminobutyric acid). There is no effective treatment for preventing or slowing this neuronal degeneration. Ciliary neurotrophic factor (CNTF) is a trophic factor for striatal neurons and therefore a potential therapeutic agent for Huntington's disease. Here we evaluate CNTF as a neuroprotective agent in a nonhuman primate model of Huntington's disease. We gave cynomolgus monkeys intrastriatal implants of polymer-encapsulated baby hamster kidney fibroblasts that had been genetically modified to secrete human CNTF. One week later, monkeys received unilateral injections of quinolinic acid into the previously implanted striatum to reproduce the neuropathology seen in Huntington's disease. Human CNTF was found to exert a neuroprotective effect on several populations of striatal cells, including GABAergic, cholinergic and diaphorase-positive neurons which were all destined to die following administration of quinolinic acid. Human CNTF also prevented the retrograde atrophy of layer V neurons in motor cortex and exerted a significant protective effect on the GABAergic innervation of the two important target fields of the striatal output neurons (the globus pallidus and pars reticulata of the substantia nigra). Our results show that human CNTF has a trophic influence on degenerating striatal neurons as well as on critical non-striatal regions such as the cerebral cortex, supporting the idea that human CNTF may help to prevent the degeneration of vulnerable striatal populations and cortical-striatal basal ganglia circuits in Huntington's disease.

Animals↗
PubMed1997

Impaired endothelium-dependent relaxation after cardiac global ischemia and reperfusion: role of warm blood cardioplegia.

OBJECTIVES: Experiments were designed to determine whether coronary endothelial dysfunction after cardiac global ischemia and reperfusion could be prevented by warm blood cardioplegic solution. BACKGROUND: The coronary endothelium produces endothelium-derived relaxing factor (EDRF) to prevent vasospasm and thrombosis. After ischemia and reperfusion, endothelium-dependent relaxation (EDR) is diminished as a result of G-protein dysfunction. METHODS: Dogs were exposed to extracorporeal circulation in 37 degrees C (group 1) or 28 degrees C (groups 2 and 3). The heart was ischemic for 120 min while continuous warm blood cardioplegic solution (group 1) or intermittent cold (4 degrees C) crystalloid cardioplegic solution was not used in group 3 animals. The heart was then allowed to function for 60 min of reperfusion. RESULTS: Endothelium-derived relaxation in response to acetylcholine, adenosine diphosphate and sodium fluoride of the coronary rings of group 1 was significantly different from that of groups 2 and 3 but was not significantly different from that of group 4. In contrast, EDR in response to the receptor-independent calcium ionophore agonist A23187 was not significantly different between the four groups. Scanning electron microscopic studies showed that platelet adhesion and aggregation, area of microthrombi, disruption of endothelial cells and separation of the intercellular junction could be found in coronary segments of groups 2 and 3 but not in vessels of groups 1 and 4. CONCLUSIONS: These experiments suggest that cardiac global ischemia and reperfusion impair receptor-mediated release of EDRF from the coronary endothelium with G-protein dysfunction. This type of coronary endothelial dysfunction can be prevented by continuous anterograde infusion of warm blood cardioplegic solution during global ischemia.

Animals↗
PubMed1997

Telomerase and cell proliferation in mouse skin papillomas.

The progression of chemically induced mouse skin papillomas is paralleled by an increase in telomerase activity. In this study, we compared telomerase activity and rate of cell proliferation in papillomas obtained early versus late in papilloma progression. Eighteen early papillomas (after 15 wk of promotion) showed no evidence of telomerase activity, and their average cell proliferation index was 26.6% +/- 6.3. On the other hand, most of the papillomas harvested after 25 wk of promotion showed high levels of telomerase activity, but their average cell proliferation index (30.8% +/- 6.2) was not different from that of the early lesions. We concluded that there appears to be no association between the level of telomerase activity observed in mouse skin papillomas and the rate of cell proliferation of each individual tumor. Telomerase expression may indicate the existence of more abundant tumor subpopulations in advanced papillomas with proliferative potential for autonomous growth.

9,10-Dimethyl-1,2-benzanthracene↗