Biomedical subjects

J You

Publications and source records attributed to J You.

At least 55 records · Page 3Linked to original sources

PubMed2000

[Developments of high temperature Raman spectroscopic techniques].

High temperature Raman spectroscopy (HTRS) is contemporarily a useful tool to study micro-structure of various materials under high temperature. Developments of HTRS techniques are well reviewed. In order to realize the determinations of metallurgical melts, the modified instrumentation for high temperature determination purpose based on JY U1000 Raman spectrometer in Shanghai Enhanced Lab of Ferrometallurgy (SELF) is introduced. It is assembled as macro-Raman with time-resolved detect system by using pulsed Copper Vapor Laser(CVL). This is the first set of its kind in domestic and some further modifications are proposed.

Hot Temperature↗

PubMed1999

Interaction with members of the heterochromatin protein 1 (HP1) family and histone deacetylation are differentially involved in transcriptional silencing by members of the TIF1 family.

Mammalian TIF1alpha and TIF1beta (KAP-1/KRIP-1) are related transcriptional intermediary factors that possess intrinsic silencing activity. TIF1alpha is believed to be a euchromatic target for liganded nuclear receptors, while TIF1beta may serve as a co-repressor for the large family of KRAB domain-containing zinc finger proteins. Here, we report an association of TIF1beta with both heterochromatin and euchromatin in interphase nuclei. Co-immunoprecipitation of nuclear extracts shows that endogenous TIF1beta, but not TIF1alpha, is associated with members of the heterochromatin protein 1 (HP1) family. However, in vitro, both TIF1alpha and TIF1beta interact with and phosphorylate the HP1 proteins. This interaction involves a conserved amino acid motif, which is critical for the silencing activity of TIF1beta but not TIF1alpha. We further show that trichostatin A, an inhibitor of histone deacetylases, can interfere with both TIF1 and HP1 silencing. The silencing activity of TIF1alpha appears to result chiefly from histone deacetylation, whereas that of TIF1beta may be mediated via both HP1 binding and histone deacetylation.

Amino Acid Sequence↗

PubMed1999

A genetic map and recombination parameters of the human malaria parasite Plasmodium falciparum.

Genetic investigations of malaria require a genome-wide, high-resolution linkage map of Plasmodium falciparum. A genetic cross was used to construct such a map from 901 markers that fall into 14 inferred linkage groups corresponding to the 14 nuclear chromosomes. Meiotic crossover activity in the genome proved high (17 kilobases per centimorgan) and notably uniform over chromosome length. Gene conversion events and spontaneous microsatellite length changes were evident in the inheritance data. The markers, map, and recombination parameters are facilitating genome sequence assembly, localization of determinants for such traits as virulence and drug resistance, and genetic studies of parasite field populations.

Animals↗

PubMed1999

E2/E3-mediated assembly of lysine 29-linked polyubiquitin chains.

Polyubiquitin (Ub) chains linked through Lys-48-Gly-76 isopeptide bonds represent the principal signal by which substrates of the Ub-dependent protein degradation pathway are targeted to the 26 S proteasome, but the mechanism(s) whereby these chains are assembled on substrate proteins is poorly understood. Nor have assembly mechanisms or definitive functions been assigned to polyubiquitin chains linked through several other lysine residues of ubiquitin. We show that rabbit reticulocyte lysate harbors enzymatic components that catalyze the assembly of unanchored Lys-29-linked polyubiquitin chains. This reaction can be reconstituted using the ubiquitin-conjugating enzyme (E2) known as UbcH5A, a 120-kDa protein(s) that behaves as a ubiquitin-protein ligase (E3), and ubiquitin-activating enzyme (E1). The same partially purified E3 preparation also catalyzes the assembly of unanchored chains linked through Lys-48. Kinetic studies revealed a K(m) of approximately 9 microM for the acceptor ubiquitin in the synthesis of diubiquitin; this value is similar to the concentration of free ubiquitin in most cells. Similar kinetic behavior was observed for conjugation to Lys-48 versus Lys-29 and for conjugation to tetraubiquitin versus monoubiquitin. The properties of these enzymes suggest that there may be distinct pathways for ubiquitin-ubiquitin ligation versus substrate-ubiquitin ligation in vivo.

Animals↗

PubMed1999

Application of the dual-micropipet technique to the measurement of tumor cell locomotion.

The objective of this work was to characterize tumor cell locomotion in response to chemotactic stimulation using a dual-micropipet assay. The assay involves two micropipets. An individual A2058 human melanoma cell was retained, without pressure gradient, in a pipet of approximately 14 micrometers i.d. A solution of type IV collagen, chosen as the chemotactic source, was placed in another pipet (approximately 10 micrometers o.d.) with zero pressure at the pipet tip. The smaller pipet was then inserted into the larger one containing the melanoma cell. The initial chemoattractant concentration (C0) and the distance between the tip of the small pipet and the cell surface (delta) provided a gradient (C0/delta) for tumor cell locomotion toward stimulation. This novel assay provides a direct measure of cell movement: cyclic pseudopod protrusion (Lp) and subsequent cell locomotion (Lc). The influences of different adhesion substrates on cell locomotion were also studied. The peak length in Lp precedes the highest locomotion velocity (dLc/dt) by an apparent lag time. C0/delta influences pseudopod protrusion frequency (fp) and dLc/dt, but not significantly on Lp. Substrate adhesions affect dLc/dt, but apparently not Lp or fp. In conclusion, pseudopod protrusion and substrate adhesion are two necessary but mutually independent factors in tumor cell locomotion. dLc/dt correlates with changes in C0/delta, which is in significant correlation with fp but not Lp.

Chemotaxis↗

PubMed1999

TIF1gamma, a novel member of the transcriptional intermediary factor 1 family.

We report the cloning and characterization of a novel member of the Transcriptional Intermediary Factor 1 (TIF1) gene family, human TIF1gamma. Similar to TIF1alpha and TIF1beta, the structure of TIF1beta is characterized by multiple domains: RING finger, B boxes, Coiled coil, PHD/TTC, and bromodomain. Although structurally related to TIF1alpha and TIF1beta, TIF1gamma presents several functional differences. In contrast to TIF1alpha, but like TIF1beta, TIF1 does not interact with nuclear receptors in yeast two-hybrid or GST pull-down assays and does not interfere with retinoic acid response in transfected mammalian cells. Whereas TIF1alpha and TIF1beta were previously found to interact with the KRAB silencing domain of KOX1 and with the HP1alpha, MODI (HP1beta) and MOD2 (HP1gamma) heterochromatinic proteins, suggesting that they may participate in a complex involved in heterochromatin-induced gene repression, TIF1gamma does not interact with either the KRAB domain of KOX1 or the HP1 proteins. Nevertheless, TIF1gamma, like TIF1alpha and TIF1beta, exhibits a strong silencing activity when tethered to a promoter. Since deletion of a novel motif unique to the three TIF1 proteins, called TIF1 signature sequence (TSS), abrogates transcriptional repression by TIF1gamma, this motif likely participates in TIF1 dependent repression.

Amino Acid Sequence↗

PubMed1999

Image/patient registration from (partial) projection data by the Fourier phase matching method.

A technique for 2D or 3D image/patient registration, PFPM (projection based Fourier phase matching method), is proposed. This technique provides image/patient registration directly from sequential tomographic projection data. The method can also deal with image files by generating 2D Radon transforms slice by slice. The registration in projection space is done by calculating a Fourier invariant (FI) descriptor for each one-dimensional projection datum, and then registering the FI descriptor by the Fourier phase matching (FPM) method. The algorithm has been tested on both synthetic and experimental data. When dealing with translated, rotated and uniformly scaled 2D image registration, the performance of the PFPM method is comparable to that of the IFPM (image based Fourier phase matching) method in robustness, efficiency, insensitivity to the offset between images, and registration time. The advantages of the former are that subpixel resolution is feasible, and it is more insensitive to image noise due to the averaging effect of the projection acquisition. Furthermore, the PFPM method offers the ability to generalize to 3D image/patient registration and to register partial projection data. By applying patient registration directly from tomographic projection data, image reconstruction is not needed in the therapy set-up verification, thus reducing computational time and artefacts. In addition, real time registration is feasible. Registration from partial projection data meets the geometry and dose requirements in many application cases and makes dynamic set-up verification possible in tomotherapy.

Algorithms↗

PubMed1999

Chromosome size-dependent control of meiotic reciprocal recombination in Saccharomyces cerevisiae: the role of crossover interference.

In the yeast Saccharomyces cerevisiae, small chromosomes undergo meiotic reciprocal recombination (crossing over) at rates (centimorgans per kilobases) greater than those of large chromosomes, and recombination rates respond directly to changes in the total size of a chromosomal DNA molecule. This phenomenon, termed chromosome size-dependent control of meiotic reciprocal recombination, has been suggested to be important for ensuring that homologous chromosomes cross over during meiosis. The mechanism of this regulation was investigated by analyzing recombination in identical genetic intervals present on different size chromosomes. The results indicate that chromosome size-dependent control is due to different amounts of crossover interference. Large chromosomes have high levels of interference while small chromosomes have much lower levels of interference. A model for how crossover interference directly responds to chromosome size is presented. In addition, chromosome size-dependent control was shown to lower the frequency of homologous chromosomes that failed to undergo crossovers, suggesting that this control is an integral part of the mechanism for ensuring meiotic crossing over between homologous chromosomes.

Chromosomes, Fungal↗

PubMed1999

Enhancement of transfection efficiency by protamine in DDAB lipid vesicle-mediated gene transfer.

We have previously developed a simple gene transfection procedure mediated by cationic lipid vesicles for animal cells, in which a commercially available cationic surfactant, dimethyldioctadecyl ammonium bromide (DDAB), was used for making lipid vesicles. In the present study, we examined enhancement of transfection efficiency for this method by adding protamine to plasmid DNA solution before the formation of DNA/lipid vesicle complexes. Both free-base protamine and protamine sulfate provided enhanced transfection efficiency and expression level, but the optimal amount of the two protamines was different. The enhancement in transfection efficiency and expression level by protamines was observed in all the cell lines (COS-7, Hela, NIH3T3, MDCK, and BHK-21C13) and all the plasmids (pCMVbeta, pmiwZ, and pCH110) tested. The enhancement in both transfection efficiency and expression level was at most 20-fold compared with that using only DDAB lipid vesicles. Protamines seemed to protect DNA from degradation by DNase and promote DNA delivery into a nucleus.

3T3 Cells↗

PubMed1999

A unified reconstruction framework for both parallel-beam and variable focal-length fan-beam collimators by a Cormack-type inversion of exponential radon transform.

A variety of inversions of exponential Radon transform has been derived based on the circular harmonic transform in Fourier space by several research groups. However, these inversions cannot be directly applied to deal with the reconstruction for fan-beam or variable-focal-length fan-beam collimator geometries in single photon emission computed tomography (SPECT). In this paper, we derived a Cormack-type inversion of the exponential Radon transform by employing the circular harmonic transform directly in the projection space and the image space instead of the Fourier space. Thus, a unified reconstruction framework is established for parallel-, fan-, and variable-focal-length fan-beam collimator geometries. Compared to many existing algorithms, the presented one greatly mitigates the difficulty of image reconstruction due to the complicated collimator geometry and significantly reduces the computational burden of the special functions, such as Chebyshev or Bessel functions. By the well-established fast-Fourier transform (FFT), our algorithm is very efficient, as demonstrated by several numerical simulations.

Algorithms↗

PubMed1999

Functional heterogeneity of endothelial P2 purinoceptors in the cerebrovascular tree of the rat.

The effects of stimulating P2Y1 or P2Y2 purinoceptors on the endothelium of isolated middle cerebral arteries (MCAs), third-order branches of the MCA (bMCAs), and penetrating arterioles (PAs) of the rat were studied. After pressurization and development of spontaneous tone (25% contraction), resting diameters for MCAs, bMCAs, and PAs were 203 +/- 5 (n = 50), 99 +/- 2 (n = 42), and 87 +/- 2 micron (n = 53), respectively. Luminal application of the P2Y1-selective agonist 2-methylthioadenosine 5'-triphosphate elicited dose-dependent dilations (or loss of intrinsic tone) in MCAs but not in bMCAs or PAs. The dilation in MCAs was completely blocked by removal of the endothelium or by nitro-L-arginine methyl ester (10(-5) M), an inhibitor of NO synthase. Luminal application of the P2Y2-selective agonist ATP elicited dilations in MCAs, bMCAs, and PAs. Removal of the endothelium abolished the dilations in all vessel groups. Dilations in MCAs have been shown to involve both NO and endothelium-derived hyperpolarizing factor (EDHF). The dilations in bMCAs and PAs had a minor NO component and prominent EDHF component; that is, 1) the dilations to ATP were not diminished by the combined inhibition of NO synthase and cyclooxygenase, 2) the dilations were accompanied by significant hyperpolarizations of the vascular smooth muscle (approximately 15 mV), and 3) the dilations were completely abolished by the calcium-activated potassium channel blocker charybdotoxin. We concluded that the role of NO in purinoceptor-induced dilations diminishes along the cerebrovascular tree in the rat, whereas the role of EDHF becomes more prominent.

Adenosine Triphosphate↗

PubMed1999

P2u receptor-mediated release of endothelium-derived relaxing factor/nitric oxide and endothelium-derived hyperpolarizing factor from cerebrovascular endothelium in rats.

BACKGROUND AND PURPOSE: Stimulation of P2u purinoceptors by UTP on endothelium dilates the rat middle cerebral artery (MCA) through the release of endothelium-derived relaxing factor/nitric oxide (EDRF/NO) and an unknown relaxing factor. The purpose of this study was to determine whether this unknown relaxing factor is endothelium-derived hyperpolarizing factor (EDHF). METHODS: Rat MCAs were isolated, cannulated, pressurized, and luminally perfused. UTP was added to the luminal perfusate to elicit dilations. RESULTS: Resting outside diameter of the MCAs in one study was 209+/-7 micrometer (n=10). The MCAs showed concentration-dependent dilations with UTP administration. Inhibition of NO synthase with NG-nitro-L-arginine methyl ester (L-NAME) (1 micromol/L to 1 mmol/L) did not diminish the maximum response to UTP but did shift the concentration-response curve to the right. Scavenging NO with hemoglobin (1 or 10 micromol/L) or inhibition of guanylate cyclase with ODQ (1 or 10 micromol/L) had effects on the UTP-mediated dilations similar to those of L-NAME. In the presence of L-NAME, dilations induced by 10 micromol/L UTP were accompanied by 13+/-2 mV (P<0.009) hyperpolarization of the vascular smooth muscle membrane potential (-28+/-2 to -41+/-1 mV). Iberiotoxin (100 nmol/L), blocker of the large-conductance calcium-activated K channels, sometimes blocked the dilation, but its effects were variable. Charybdotoxin (100 nmol/L), also a blocker of the large-conductance calcium-activated K channels, abolished the L-NAME-insensitive component of the dilation to UTP. CONCLUSIONS: Stimulation of P2u purinoceptors on the endothelium of the rat MCA released EDHF, in addition to EDRF/NO, and dilated the rat MCA by opening an atypical calcium-activated K channel.

4-Aminopyridine↗

PubMed1999

Construct for high-level expression and low misincorporation of lysine for arginine during expression of pET-encoded eukaryotic proteins in Escherichia coli.

The arginine codon AGA is rarely used in E. coli but is common in eukaryotic genes. Prior studies have shown that the low level of tRNA(UCUArg) can lead to low expression and misincorporation of lysine for arginine, during expression of genes containing AGA codons in E. coli. The chloramphenicol-selectable plasmid pJY2 is designed to facilitate the expression of such genes cloned into pET vectors: it encodes T7 lysozyme (to depress constitutive expression of the cloned gene) and tRNA(UCUArg) (to suppress lysine misincorporation at AGA codons). Using pJY2, we observed robust and translationally faithful expression of mutant ubiquitin genes in which 14% (11 out of 76) of the total codons were AGA. Competent BL21(DE3)pJY2 cells can be used to suppress lysine misincorporation and achieve high-level expression of pET-encoded target genes without modification of AGA codons in the target gene sequence.

Arginine↗

PubMed1999

[The establishment and application of genomic DNA extraction from peripheral blood].

OBJECTIVE: To establish the method of genomic DNA extraction. METHODS: Genomic DNA was directly extracted from peripheral blood with the use of potassium iodide. RESULTS: The genomic DNA extracted with potassium iodide was of large molecular weight. A(260)/A(280) and A(260)/ A(230) were 1.85 and 2.2 respectively. The extraction efficiency was greater than 90%. The result of PCR for the DNA was good. CONCLUSION: The method is simple, quick, economical and could be used for studying numbers of clinical samples.

DNA↗

PubMed1999

[The role of NF-kappa B in the TNF-alpha-induced endothelial cell apoptosis].

OBJECTIVE: To investigate whether TNF-alpha could activate the signaling pathway-NF-kappa B/I-kappa B alpha required for the expression of inducible nitric oxide synthase (iNOS) to induce endothelial cell apoptosis by nitric oxide (NO). METHODS: Aminoguanidine (AG, an inhibitor of iNOS) and Pyrrolidine dithiocarbamate (PDTC, an inhibitor of NF-kappa B) were evaluated for their influence on the apoptosis of cultured bovine pulmonary artery endothelial cells (BPAEC) induced by TNF-alpha. Apoptosis was confirmed by morphology, ultrastructural observation, agarose gel electrophoresis of DNA, and percentage of DNA fragmentation. Western blot analyses were used to detect the expression of iNOS and the levels of I-kappa B alpha in BPAEC exposed to TNF-alpha. Electrophoretic mobility shift assays (EMSA) were used to determinate the activity of NF-kappa B in BPAEC. RESULTS: TNF-alpha induced BPAEC apoptosis (18.0 +/- 4.3)% in a concentration- and time-dependent manner. Both AG and PDTC attenuated the apoptosis of BPAEC induced by TNF-alpha, with peventages of DNA fragmentation in 2, 4, 10 mmol/L AG being (10.0 +/- 2.2)%, (7.8 +/- 1.2)% and (8.2 +/- 1.3)% vespectively. In BPAEC after TNF-alpha exposure, Western blot analyses revealed the expression of iNOS and the decrease in I-kappa B alpha. EMSA demonstrated an impermanency increase in NF-kappa B binding activity. CONCLUSIONS: TNF-alpha can induce endothelial cell apoptosis by NO, which is produced by increasing iNOS expression and activating the signal pathway-NF-kappa B.

Animals↗

PubMed1999

[Expression of membrane-type I matrix metalloproteinase and its correlation with gelatinase A activation in cancer cell lines].

OBJECTIVE: To study the expression of membrane-type I matrix metalloproteinase (MT1-MMP) and its correlation with gelatinase A (MMP2) activation. METHODS: Reverse transcription PCR, northern blots and western blots were employed to detect the expression levels of MT1-MMP mRNA and MMP2 protein in 4 human melanoma cell lines, 2 human lung cancer cell lines and 2 human prostate cancer cell lines. RESULTS: All the cancer cell lines expressed MT1-MMP mRNA, and the MT1-MMP mRNA level in the 4 melanoma cell lines was significantly higher than that in the lung and prostate cancer cell lines. Activated MMP2 proteins were only detected in the melanoma cell lines, whereas PG, a lung cancer cell line, which secreted pro-MMP2 and expressed low level of MT1-MMP, could not produce activated MMP2. CONCLUSIONS: The level of MT1-MMP expression was highly associated with MMP2 activation.

Blotting, Northern↗

PubMed1999

[Isolation and characterization of human prostate cancer cell subclones with different metastatic potential].

OBJECTIVE: To isolate and characterize cancer cell subclones with different metastatic potential from human metastatic prostate cancer cell line (PC-3M). METHODS: Using limited dilution, in vitro growth, Matrigel invasion assay, soft agar cloning and in vivo tumorigenicity and spontaneous metastasis assay in nude mice, were isolated and characterized four subclones with different metastatic potential. RESULTS: Four subclones derived from PC-3M were 1E8, 2E7, 2B6, 2B4. Each subclone exhibited a different metastatic potential when inoculated into nude mice. Among these subclones, 1E8 expressed as the highly metastatic phenotype, with 100% metastasis frequency 5 weeks after subcutaneous inoculation into nude mice, whereas 2B4 was not metastatic. In vitro 1E8 was found to be the most highly invasive cell line in Matrigel invasive assay (98 +/- 24) and had the most clones in soft agar cloning assay (265 +/- 39) while 2B4 was found to have the least invasive abilities (12 +/- 4) and the least clones (137 +/- 14). CONCLUSION: Successful establishment of these subclones with different metastatic potential may be valuable for further study on the molecular mechanisms of cancer metastasis and cloning of cancer metastasis-related genes.

Animals↗

PubMed1999

[Application of supercritical fluid extraction (SFE) on environmental analysis].

In this paper, publications from 1993 are classified according to the type of matrix from which the environmental analytes are extracted. Although the most applications of SFE are used for the solid, some analysts have concerned with aqueous and gaseous media. Various aspects of investigation of environmental potential carcinogen, such as, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, dioxins, pesticides, phenols and other hazardous and toxic pollutants in environmental samples have been reviewed with 55 references. The coupled techniques of SFE have also been introduced in this paper.

Air Pollutants↗