PubMed Health⌕ Search

Biomedical subjects

C Yu

Publications and source records attributed to C Yu.

At least 127 records · Page 7Linked to original sources
PubMed2000

Role of Niemann-Pick type C1 protein in intracellular trafficking of low density lipoprotein-derived cholesterol.

Niemann-Pick type C (NPC) is a disease that affects intracellular cholesterol-trafficking pathways. By cloning the hamster ortholog of NPC1, we identified the molecular lesions in two independently isolated Chinese hamster ovary cell mutants, CT60 and CT43. Both mutants lead to premature translational terminations of the NPC1 protein. Transfecting hamster NPC1 cDNA complemented the defects of the mutants. Investigation of the CT mutants, their parental cells, and an NPC1-stable transfectant allow us to present evidence that NPC1 is involved in a post-plasma membrane cholesterol-trafficking pathway. We found that the initial movement of low density lipoprotein (LDL)-derived cholesterol to the plasma membrane (PM) did not require NPC1. After reaching the PM and subsequent internalization, however, cholesterol trafficking back to the PM did involve NPC1. Both LDL-derived cholesterol and cholesterol originating from the PM accumulated in a dense, intracellular compartment in the CT mutants. Cholesterol movement from this compartment to the PM or endoplasmic reticulum was defective in the CT mutants. Our results functionally distinguish the dense, intracellular compartment from the early endocytic hydrolytic organelle and imply that NPC1 is involved in sorting cholesterol from the intracellular compartment back to the PM or to the endoplasmic reticulum.

Amino Acid Sequence↗
PubMed2000

Transport and pharmacological properties of nine different human Na, K-ATPase isozymes.

Na,K-ATPase plays a crucial role in cellular ion homeostasis and is the pharmacological receptor for digitalis in man. Nine different human Na,K-ATPase isozymes, composed of 3 alpha and beta isoforms, were expressed in Xenopus oocytes and were analyzed for their transport and pharmacological properties. According to ouabain binding and K(+)-activated pump current measurements, all human isozymes are functional but differ in their turnover rates depending on the alpha isoform. On the other hand, variations in external K(+) activation are determined by a cooperative interaction mechanism between alpha and beta isoforms with alpha2-beta2 complexes having the lowest apparent K(+) affinity. alpha Isoforms influence the apparent internal Na(+) affinity in the order alpha1 > alpha2 > alpha3 and the voltage dependence in the order alpha2 > alpha1 > alpha3. All human Na,K-ATPase isozymes have a similar, high affinity for ouabain. However, alpha2-beta isozymes exhibit more rapid ouabain association as well as dissociation rate constants than alpha1-beta and alpha3-beta isozymes. Finally, isoform-specific differences exist in the K(+)/ouabain antagonism which may protect alpha1 but not alpha2 or alpha3 from digitalis inhibition at physiological K(+) levels. In conclusion, our study reveals several new functional characteristics of human Na,K-ATPase isozymes which help to better understand their role in ion homeostasis in different tissues and in digitalis action and toxicity.

Animals↗
PubMed2000

Analysis of a commercial preparation of erythromycin estolates by tandem mass spectrometry and high performance liquid chromatography/electrospray ionization tandem mass spectrometry using an ion trap mass spectrometer.

Because of the lack of a UV chromophore and their much smaller abundances in comparison with the major component, the minor components in erythromycin estolate preparations are difficult to analyze by high performance liquid chromatography ultraviolet (HPLC-UV). Tentative assignment of the major and minor components can be achieved with the combination of full scan and ZoomScan using an ion trap mass spectrometer. Tandem mass spectrometry (MS/MS) provided an effective method to quickly identify most components without chromatographic separation, and all the related compounds, except the isobaric pair ECE and PdMeEA, could be identified in this way. The best result was obtained by using liquid chromatography/tandem mass spectrometry (LC/MS/MS) operated in selected reaction monitoring mode. The major compound, the estolate of erythromycin A (EAE), and seven other minor components, could be separated and identified, with semiquantitative estimates of relative concentrations.

Anti-Bacterial Agents↗
PubMed2000

Towards stationary phases for chromatography on a microchip: molded porous polymer monoliths prepared in capillaries by photoinitiated in situ polymerization as separation media for electrochromatography.

Photoinitiated free radical polymerization has been used for the preparation of porous polymer monoliths within UV transparent fused silica capillaries and quartz tubes. These formats were used as models for the preparation of the separation media within channels of microfabricated devices. A mixture of ethylene dimethacrylate, butyl methacrylate, and 2-acrylamido-2-methyl-1-propanesulfonic acid was polymerized in the presence of a porogenic solvent consisting of 1-propanol, 1,4-butanediol, and water at room temperature under UV irradiation. Modification of the porogen composition enables the tailoring of pore size within the broad range from ca. 100 to 4000 nm. Scanning electron micrographs confirmed the homogeneity of the porous structure of the materials prepared, even in a quartz tube with a diameter as large as 4 mm. Separation properties of the resulting capillary columns were tested in capillary electrochromatography (CEC) mode using a mixture of thiourea and eight aromatic compounds. Plate number as high as 210 000 plates/m were found for a capillary column with optimized porous properties. The monolithic columns were also able to separate mixtures of peptides.

Animals↗
PubMed2000

Expression of collagen type I, II and III in loose body of osteoarthritis.

The expression of collagen type I, II, and III was investigated to evaluate phenotypic change in chondrocytes in loose bodies related to osteoarthritis. We assessed collagen type I, II, and III production in loose bodies from knee joints of ten osteoarthritic patients, using an immunohistochemical method with monoclonal antibodies. Collagen type III expression was identified in all ten loose bodies and was mainly located in cartilage, including chondrocytes and matrices, as well as in a layer of fibroid tissue on the surface. No positive signal for collagen type III was observed in necrotic osteocytes. There was weakly positive staining for collagen type I in chondrocytes. No positive staining for collagen type II could be seen in the cartilage of loose bodies. Cartilage from the non-osteoarthritic knee joints of four people was negative for the expression of collagen type I and III, and positive for the expression of collagen type II. Collagen type I and III expression suggested the dedifferentiation status of chondrocytes in loose bodies.

Adult↗
PubMed2000

Fusion induced aggregation of model vesicles studied by dynamic and static light scattering.

Membrane fusion is a key step in the virus mediated cell fusion. The vesicular dispersion serves as a model system to study the membrane fusion. We employed dynamic and static light scattering to study the fusion of phosphatidylcholine vesicles in the presence of model fusion peptide fragments from the hemagglutinin HA2 protein. The fusion-induced aggregation under the present experimental setup exhibited strong pH dependence, similar to the parental viral protein. Replacement of the glycine residue at the extreme amino terminus by glutamic acid (G1E) abolished fusion activity. The average molecular mass and diameter of vesicular dispersion obtained from static and dynamic light scattering measurements respectively at neutral and acidic pH showed about three fold increase in acidic solution containing wild type fusion peptide. The light scattering data are consistent with lipid mixing results. The present work demonstrates the utility of light scattering as a facile means to monitor the fusion process.

Amino Acid Sequence↗
PubMed2000

Genetic strategies to enhance penicillin acylase production in Escherichia coli.

We demonstrated the improvement of penicillin acylase (PAC) production by optimization of the host/vector system using genetic engineering strategies. Several expression plasmids with improved efficiency for the transcription of the pac gene and/or translation of the pac mRNA were constructed. Mutant strains, isolated by a novel screening method, were effective for use as the expression host to produce PAC. The feasibility of using the mutant strains harboring a selection of expression plasmids for the production of PAC was evaluated. The effect of the mutation(s) resulting in the improved PAC producing ability was characterized. While the production of PAC was significantly enhanced using the optimized host/vector system, the formation of PAC inclusion bodies was shown to be another step limiting the production of recombinant PAC.

Journal Article↗
PubMed2000

Evidence for migration of donor bone marrow stromal cells into recipient thymus after bone marrow transplantation plus bone grafts: A role of stromal cells in positive selection.

Intrathymic T-cell differentiation is characterized by two selection events: positive and negative selection. It has been shown that thymic epithelial cells in the cortex are involved in the positive selection, while macrophages and dendritic cells, derived from hemopoietic stem cells, are involved in the negative selection. Here we investigate whether donor-derived bone marrow stromal cells can migrate into the thymus and participate there in positive selection after bone marrow transplantation plus bone grafts (to recruit bone marrow stromal cells). Allogeneic bone marrow transplantation with or without bone grafts was carried out in the [C57BL/6-->C3H] combination. Fluorescence-activated cell sorter analyses of recipient thymic adherent cells showed that donor-type bone marrow stromal cells exist in the thymus of mice that received bone marrow plus bone grafts but not in the mice that received bone marrow cells alone. Histological examination using confocal microscopy also confirmed the existence of donor-type stromal cells in the thymus of mice that received bone marrow cells plus bones. Both T-cell proliferation and plaque-forming cell assays indicated that the T cells of such mice show donor-type major histocompatibility complex-restriction. These findings strongly suggest that stromal cells can migrate from the bone marrow to the thymus, where they participate in the positive selection of thymocytes.

Animals↗
PubMed2000

Fetal renal volume in normal gestation: a three-dimensional ultrasound study.

To establish a reference chart of fetal kidneys in normal pregnancy, we performed a prospective and cross-sectional study. A total of 152 singleton fetuses ranging between 20 and 40 weeks' gestation and meeting the criteria of normal pregnancies were included. Three-dimensional ultrasound (3-D US) was used to measure the fetal renal volume. Our results revealed that both renal volumes are highly correlated with the fetal gestational age. Using gestational age (GA) as the independent variable and right renal volume (RRV) as the dependent variable, the best-fit regression equation was RRV (mL)=0.74053xGA (week)-13.318 (r = 0.89, p < 0.001). Similarly, the best-fit equation for the left renal volume (LRV) was LRV (mL)=0. 76093xGA (week)-13.421 (r = 0.86, p < 0.001). The normal growth centiles of both kidneys were established based on these two equations. There were no significant differences of the volumes between bilateral kidneys. In conclusion, our data of fetal renal volumes assessed by 3-D US may serve as a reference in evaluating fetal renal growth.

Cross-Sectional Studies↗
PubMed2000

Hereditary haemorrhagic telangiectasia: hepatic lesions demonstrated with colour Doppler and power Doppler sonography.

Visceral vascular abnormalities in patients with hereditary haemorrhagic teleangiectasia (HHT) include teleangiectasis, arteriovenous fistula (AVF), and aneurysms. Sonographic findings and colour Doppler ultrasound (CDU) features of HHT in the liver have been described in a few reports. Presented here is the CDU finding of liver with HHT involvement in two patients, aged 52 and 29 years, respectively. Power Doppler sonography was also performed in the first patient in whom both CDU and power Doppler sonography could demonstrate the abnormal vascular channels in the liver. In the second patient, aneurysms in the left hepatic lobe and of the splenic artery were noted. Angiography was performed in both patients and confirmed the abnormal vascular lesions demonstrated by CDU and/or power Doppler sonographic findings.

Adult↗
PubMed2000

Substituted benz[a]acridines and benz[c]acridines as mammalian topoisomerase poisons.

Coralyne and several other synthetic benzo[a,g]quinolizium derivatives related to protoberberine alkaloids have exhibited activity as topoisomerase poisons. These compounds are characterized by the presence of a positively charged iminium group, which has been postulated to be associated with their pharmacological properties. The objective of the present study was to devise stable noncharged bioisosteres of these compounds. Several similarly substituted benz[a]acridine and benz[c]acridine derivatives were synthesized and their relative activity as topoisomerase poisons was determined. While the benz[c]acridine derivatives evaluated as part of this study were devoid of topoisomerase poisoning activity, several dihydrobenz[a]acridines were able to enhance DNA cleavage in the presence of topo I. In contrast to certain protoberberine derivatives that did exhibit activity as topo II poisons, none of the benz[a]acridines derivatives enhanced DNA cleavage in the presence of topo II. Among the benz[a]acridines studied, 5,6-dihydro-3,4-methylenedioxy-9,10-dimethoxybenz[a]acridine, 13e, was the most potent topo I poison, with comparable potency to coralyne. These data suggest that heterocyclic compounds structurally related to coralyne can exhibit potent topo I poisoning activity despite the absence of an iminium cation within their structure. In comparison to coralyne or other protoberberine derivatives, these benz[a]acridine derivatives possess distinctly different physicochemical properties and represent a novel series of topo I poisons.

Acridines↗
PubMed2000

Surround modulation in human vision unmasked by masking experiments.

The responses of neurons in cat and monkey primary visual cortex are modulated by stimuli outside the classical receptive field. Here we report psychophysical evidence from masking experiments for two distinct types of surround modulation, one narrowly tuned to iso-orientation (stimuli with center and surround at the same orientation) and the other broadly tuned to cross-orientation (center and surround at perpendicular orientations). Surround modulation at iso- and cross-orientations showed distinct contrast dependencies, and high-contrast cross-oriented surrounds were able to completely eliminate masking. Surround modulation was modeled by subtracting divisive inhibition that raised the gain of spatial filters.

Adult↗
PubMed2000

Contribution of Are1p and Are2p to steryl ester synthesis in the yeast Saccharomyces cerevisiae.

In the yeast Saccharomyces cerevisiae, two acyl-CoA:sterol acyltransferases (ASATs) that catalyze the synthesis of steryl esters have been identified, namely Are2p (Sat1p) and Are1p (Sat2p). Deletion of either ARE1 or ARE2 has no effect on cell viability, and are1are2 double mutants grow in a similar manner to wild-type despite the complete lack of cellular ASAT activity and steryl ester formation [Yang, H., Bard, M., Bruner, D. A., Gleeson, A., Deckelbaum, R. J., Aljinovic, G., Pohl, T. M., Rothstein, R. & Sturley, S. L. (1996) Science 272, 1353-1356; Yu, C., Kennedy, J., Chang, C. C. Y. & Rothblatt, J. A. (1996) J. Biol. Chem. 271, 24157-24163]. Here we show that both Are2p and Are1p reside in the endoplasmic reticulum as demonstrated by measuring ASAT activity in subcellular fractions of are1 and are2 deletion strains. This localization was confirmed by fluorescence microscopy using hybrid proteins of Are2p and Are1p fused to green fluorescent protein (GFP). Lipid analysis of are1 and are2 deletion strains revealed that Are2p and Are1p utilize sterol substrates in vivo with different efficiency; Are2p has a significant preference for ergosterol as a substrate, whereas Are1p esterifies sterol precursors, mainly lanosterol, as well as ergosterol. The specificity towards fatty acids is similar for both isoenzymes. The lack of steryl esters in are1are2 mutant cells is largely compensated by an increased level of free sterols. Nevertheless, terbinafine, an inhibitor of ergosterol biosynthesis, inhibits growth of are1are2 cells more efficiently than growth of wild-type. In a growth competition experiment are1are2 cells grow more slowly than wild-type after several rounds of cultivation, suggesting that Are1p and Are2p or steryl esters, the product formed by these two enzymes, are more important in the natural environment than under laboratory conditions.

Acyltransferases↗
PubMed2000

Radiosurgical salvage therapy for patients presenting with recurrence of metastatic disease to the brain.

OBJECTIVE: Radiosurgery has emerged as an important modality in the management of metastatic disease to the brain. A number of groups have published results suggesting that high local control rates can be achieved, with improvements in overall survival that rival the results of open surgical treatment. Typically, however, whole-brain radiotherapy has been used in the salvage therapy of patients who have undergone previous craniotomy or radiosurgery. We describe our experience with radiosurgical salvage in this group of patients. METHODS: From August 1994 to February 1999, 190 patients with brain metastasis were treated with gamma unit radiosurgery at our institution. A subset of 45 patients, who underwent radiosurgical salvage for new tumors in a region remote from an initially treated tumor, form the population base for this study. The usual criteria for repeat treatment were recurrence with five or fewer discrete lesions outside of the previously treated radiosurgical volume and Karnofsky Performance Scale score of at least 70. Survival and freedom from progression were measured from the time of radiosurgical treatment and were computed by the Kaplan-Meier product-limit method. Two or more curves were compared using the log-rank method. RESULTS: In this subgroup of patients, a total of 176 tumors were treated. The median time from first radiosurgical procedure to first salvage was 17.4 weeks. Median survival from the second radiosurgical intervention was 28 weeks. Of the 45 study patients, 34 patients underwent a single salvage procedure, 10 patients underwent two salvage procedures, and 1 patient had three salvage procedures. The actuarial freedom from progression for treated tumors at 52 weeks was 92.4%. Patients undergoing upfront whole-brain irradiation were less likely to require salvage therapy (P = 0.008). There were 33 deaths after salvage radiosurgery during the reporting period. Central nervous system causes accounted for 13 deaths, whereas 19 deaths resulted from systemic disease. The cause of death in one patient could not be determined. No statistically significant advantage in overall survival could be demonstrated in patients treated with whole-brain irradiation. CONCLUSION: Radiosurgical salvage represents a valuable means of treatment for central nervous system recurrence for patients who have undergone previous treatment for metastatic disease to the brain. Whole-brain irradiation may reduce the need for salvage therapy, but no advantage in overall survival could be demonstrated in this subgroup.

Brain Neoplasms↗
PubMed2000

Stereotactic radiosurgery in the treatment of metastatic disease to the brain.

OBJECTIVE: In recent years, stereotactic radiosurgery has been growing in popularity as a treatment modality for metastatic disease to the brain. The technique has advantages of reduced cost and low morbidity compared with open surgical treatment. Furthermore, it avoids the potential cognitive side effects of fractionated whole-brain radiotherapy. We undertook this study to determine the usefulness of adjuvant radiation therapy and to determine prognostic factors in patients treated with stereotactic radiosurgery. METHODS: We reviewed our series of patients with metastatic tumors treated using gamma knife stereotactic radiosurgery from August 1994 to February 1999. Nonparametric methods were used to compare treatment subgroups by demographic features including age, Karnofsky Performance Scale score, diagnosis, and systemic disease status. Univariate and multivariate analyses of survival and freedom from progression were performed using Kaplan-Meier and Cox proportional hazards regression techniques. RESULTS: This study included 190 patients harboring 431 lesions who were treated in 263 treatment sessions. The median follow-up after radiosurgery was 36 weeks for all patients. The median actuarial survival from the time of radiosurgery in all patients was 34 weeks. When patients were stratified according to tumor histology, those without melanoma had a median survival of 39 weeks, and those with melanoma had a median survival of 28 weeks. The cause of death could be determined in 122 (92%) of the patients known to have died during the data capture period. For patients harboring melanoma, death was attributable to systemic disease in 31 (47%), to central nervous system-related processes in 29 (44%), and to unknown causes in 6 (9%). For non-melanoma patients, death was attributable to systemic disease in 45 (68%), to central nervous system-related processes in 17 (26%), and to unknown causes in 4 (6%). Significantly improved survival (P = 0.002) was observed in patients with controlled systemic disease. No significant difference in survival could be ascertained for patients presenting with up to four lesions, although patients with a total tumor volume greater than 9 cc had shortened survival. No survival benefit could be demonstrated for whole-brain radiotherapy administered either concomitantly or after radiosurgery. CONCLUSION: Factors correlated with significantly improved survival included controlled systemic disease and non-melanoma histology. We found no significant survival benefit that could be discerned from adjuvant whole-brain radiotherapy in this patient group.

Adult↗
PubMed2000

In situ and interrupted-growth studies of the self-assembly of octadecyltrichlorosilane monolayers

We have examined the self-assembly process of octadecyltrichlorosilane on silicon using x-ray reflectivity. By comparing the commonly used "interrupted-growth" characterization technique with results obtained in situ, we have determined that quenching the growth and then rinsing and drying the sample introduces free area into the film, presumably by removal of non-cross-linked (physisorbed) molecules. Reintroduction of a quenched and rinsed film to solvent does not restore the thickness of the film to its previous value. We have also performed in situ growth studies over a range of concentrations. For all concentrations, we observe growth of islands of vertical molecules. The growth follows Langmuir kinetics, except at short times for low concentration solutions.

Journal Article↗