Biomedical subjects

Y Guo

Publications and source records attributed to Y Guo.

At least 145 records · Page 8Linked to original sources

PubMed2001

[Effects of matrine on the relative molecules expression of proliferation and apoptosis in K562 cells].

OBJECTIVE: To study the effects of matrine on proliferation and apoptosis of K562 cells. METHODS: Cell culture and cell count, cell cycle phases and number of apoptotic cells were analyzed by flow cytometry. Morphology of apoptotic cells was studied by electron microscopy, BCL-6 and proliferating Cell nuclear antigen (PCNA) were checked by Western blot. RESULTS: Matrine could inhibit the proliferation of K562 cells with inhibition rate of 69% after 72 hours matrine induction; number of K562 cells increased significantly in G1 phase, and number of apoptotic cell was 68% at 5th day of matrine treated cells; the typical morphology of apoptotic cells of K562 cells was observed by electron microscopy; BCL-6 was increased and PCNA was decreased in K562 cells induced by matrine. CONCLUSION: Matrine inhibited the proliferation and induced the apoptosis of K562 cells significantly, the mechanism of which might be related to cell cycle arrest and the change of PCNA and BCL-6.

Alkaloids↗

PubMed2001

[On the characteristics of velocity modulation laser spectroscopy].

The technical properties of velocity modulation laser spectroscopy were investigated using the spectrum of A2 pi u-X2 sigma g+ (2-0) band of Meinel system of N2+. The dependence of spectral intensity on discharge current, the pressure of the gas mixture and the pump velocity of the vacuum pump were studied. Making uses of the results of these studies, the sensitivity and resolution of the spectrometer are improved by above 3 times, and economized the experimental expenses can be reduced to as 3/4 time as normal, which confirms that the results of this paper is of considerable uses.

Lasers↗

PubMed2001

[Assignments of FIR-LMR spectra of CF X 2Pi (nu = 1) and MIR-LMR spectra of NO X 2Pi (nu = 1<--0)].

According to the theories of Zeeman effect and the principle of laser magnetic resonance (LMR), a method and model for the analysis of the laser magnetic resonance spectra were developed and successfully employed to assign the FIR-LMR spectra of CF X2 Pi (nu = 1) and the MIR-LMR spectra of NO X 2Pi (nu = 1<--0). This work provided an efficient and accurate approach for primary analysis of the LMR spectra of novel radicals.

English Abstract↗

PubMed2000

CARD9 is a novel caspase recruitment domain-containing protein that interacts with BCL10/CLAP and activates NF-kappa B.

BCL10/CLAP is an activator of apoptosis and NF-kappaB signaling pathways and has been implicated in B cell lymphomas of mucosa-associated lymphoid tissue. Although its role in apoptosis remains to be determined, BCL10 likely activates NF-kappaB through the IKK complex in response to upstream stimuli. The N-terminal caspase recruitment domain (CARD) of BCL10 has been proposed to function as an activation domain that mediates homophilic interactions with an upstream CARD-containing NF-kappaB activator. To identify upstream signaling partners of BCL10, we performed a mammalian two-hybrid analysis and identified CARD9 as a novel CARD-containing protein that interacts selectively with the CARD activation domain of BCL10. When expressed in cells, CARD9 binds to BCL10 and activates NF-kappaB. Furthermore, endogenous CARD9 is found associated with BCL10 suggesting that both proteins form a pre-existing signaling complex within cells. CARD9 also self-associates and contains extensive coiled-coil motifs that may function as oligomerization domains. We propose here that CARD9 is an upstream activator of BCL10 and NF-kappaB signaling.

Adaptor Proteins, Signal Transducing↗

PubMed2000

Functional analysis of human ornithine decarboxylase alleles.

It has been known for > 10 years that there are two alleles of the human ornithine decarboxylase (ODC) gene, defined by a polymorphic PstI RFLP in intron 1. We have sequenced a large portion of each of the two alleles, including some of the 5' promoter region, exon 1, intron 1, and exon 2, and determined that a single nucleotide polymorphism at base +317 (relative to transcription start site) is responsible for the presence or absence of the PstI restriction site. We have developed two genotyping assays, a PCR-RFLP assay and a high-throughput TaqMan-based method, and determined the ODC genotype distribution in >900 North American DNA samples. On the basis of its location between two closely spaced Myc/Max binding sites (E-boxes), we speculated that the single nucleotide polymorphism at base +317 could have functional significance. Results of transfection assays with allele-specific reporter constructs support this hypothesis. The promoter/regulatory region derived from the minor ODC allele (A allele) was more effective in driving luciferase expression in these assays than the identical region from the major allele (G allele). Our results suggest that individuals homozygous for the A allele may be capable of greater ODC expression after environmental exposures, especially those that up-regulate c-MYC expression.

3T3 Cells↗

PubMed2000

G protein-coupled receptor-induced sensitization of phospholipase C stimulation by receptor tyrosine kinases.

Activation of stably expressed M(2) and M(3) muscarinic acetylcholine receptors (mAChRs) as well as of endogenously expressed lysophosphatidic acid and purinergic receptors in HEK-293 cells can induce a long lasting potentiation of phospholipase C (PLC) stimulation by these and other G protein-coupled receptors (GPCRs). Here, we report that GPCRs can induce an up-regulation of PLC stimulation by receptor tyrosine kinases (RTKs) as well and provide essential mechanistic characteristics of this sensitization process. Pretreatment of HEK-293 cells for 2 min with carbachol, a mAChR agonist, lysophosphatidic acid, or ATP, followed by agonist washout, strongly increased (by 2-3-fold) maximal PLC stimulation (measured >/=40 min later) by epidermal growth factor and platelet-derived growth factor, but not insulin, and largely enhanced PLC sensitivity to these RTK agonists. The up-regulation of RTK-induced PLC stimulation was cycloheximide-insensitive and was observed for up to approximately 90 min after removal of the GPCR agonist. Sensitization of receptor-induced PLC stimulation caused by prior M(2) mAChR activation was fully prevented by pertussis toxin and strongly reduced by expression of Gbetagamma scavengers. Furthermore, inhibition of conventional protein kinase C (PKC) isoenzymes and chelation of intracellular Ca(2+) suppressed the sensitization process, while overexpression of PKC-alpha, but not PKC-betaI, further enhanced the M(2) mAChR-induced sensitization of PLC stimulation. None of these treatments affected acute PLC stimulation by either GPCR or RTK agonists. Taken together, short term activation of GPCRs can induce a strong and long lasting sensitization of PLC stimulation by RTKs, a process apparently involving G(i)-derived Gbetagammas as well as increases in intracellular Ca(2+) and activation of a PKC isoenzyme, most likely PKC-alpha.

Adenosine Triphosphate↗

PubMed2000

The shape of 433 eros from the NEAR-shoemaker laser rangefinder

Measurements from the Near Earth Asteroid Rendezvous (NEAR)-Shoemaker Laser Rangefinder (NLR) indicate that asteroid 433 Eros is a consolidated body with a complex shape dominated by collisions. The offset between the asteroid's center of mass and center of figure indicates a small deviation from a homogeneous internal structure that is most simply explained by variations in mechanical structure. Regional-scale relief and slope distributions show evidence for control of topography by a competent substrate. Impact crater morphology is influenced by both gravity and structural control. Small-scale topography reveals ridges and grooves that may be generated by impact-related fracturing.

Journal Article↗

PubMed2000

Promoter opening by sigma(54) and sigma(70) RNA polymerases: sigma factor-directed alterations in the mechanism and tightness of control.

Transcription control at the melting step is not yet understood. Here, band shift, cross-linking, and transcription experiments on diverse DNA probes were used with two bacterial RNA polymerase holoenzymes that differ in how they regulate melting. Data indicated that both sigma(54) and sigma(70) holoenzymes assume a default closed form that cannot establish single-strand binding. Upon activation the enzymes are converted to an open form that can bind simultaneously to the upstream fork junction and to the melted transcription start site. The key difference is that sigma(54) imposes tighter regulation by creating a complex molecular switch at -12/-11; the current data show that this switch can be thrown by activator. In this case an ATP-bound enhancer protein causes sigma(54) to alter its cross-linking pattern near -11 and also causes a reorganization of holoenzyme: DNA interactions, detected by electrophoretic mobility-shift assay. At a temperature-dependent sigma(70) promoter, elevated temperature alone can assist in triggering conformational changes that enhance the engagement of single-strand DNA. Thus, the two sigma factors modify the same intrinsic opening pathway to create quite different mechanisms of transcriptional regulation.

Adenine↗

PubMed2000

Synthetic inhibitor of matrix metalloproteases decreases tumor growth and metastases in a syngeneic model of rat prostate cancer in vivo.

Members of the matrix metalloprotease (MMP) family are implicated in the progression of several malignancies including prostate cancer due to their ability to break down extracellular matrix (ECM) components. In this study, we have evaluated the ability of a synthetic MMP inhibitor (A-177430) to block tumor growth and metastases in a syngeneic model of rat prostate cancer. In an in vitro substrate assay, A-177430 exhibited nanomolar potency (IC(50) 2-6 nM) against the enzymatic activity of several MMPs. For in vivo studies, male Copenhagen rats were injected s.c. with Mat Ly Lu rat prostate cancer cells (1 x 10(6) cells ) into the right flank and animals were administered i.p.with different doses (10-100 mg/kg per day) of A-177430 for 16 days. Administration of A-177430 resulted in a dose-dependent decrease in tumor volume as compared to a control group of animals receiving vehicle alone. The maximum dose (100 mg/kg per day) of A-177430 exhibited complete arrest in tumor growth and prevented the development of macroscopic tumor metastases to lungs without exhibiting any noticeable side effects. Histologic examination of primary tumors from experimental animals showed extensive tumor necrosis and decreased tumor angiogenesis as determined by factor VIII staining of primary tumors following A-177430 treatment. These primary tumors from experimental animals also exhibited a significant increase in tumor cell DNA fragmentation as determined by TUNEL assay. Collectively, these results demonstrate the ability of MMP inhibitors to block tumor growth and metastases by blocking ECM degradation and by inhibiting tumor angiogenesis and promotion of prostate cancer cell apoptosis in vivo.

Animals↗

PubMed2000

The unusually stable coiled-coil domain of COMP exhibits cold and heat denaturation in 4-6 M guanidinium chloride

A high thermal stability is observed for the five-stranded alpha-helical coiled-coil domain of cartilage oligomeric matrix protein COMP. It does not unfold in non-denaturing buffer between 0 and 100 degrees C and thermal denaturation is only achieved at high concentrations of guanidinium chloride (4-6 M). In these solutions the protein structure is lost at decreasing (cold denaturation) and increasing temperatures (heat denaturation). In the cold denaturation region, the melting profile showed deviations from the theory of Privalov et al. [P.L. Privalov, V. Griko Yu, S. Venyaminov, V.P. Kutyshenko, Cold denaturation of myoglobin, J. Mol. Biol. 190 (1986) 487-498] probably due to deviations from a two-state mechanism. High thermal stability as well as cold and heat denaturation was also observed for a mutant of the coiled-coil domain of COMP in which glutamine 54 was replaced by isoleucine but it still forms pentamer. The melting temperatures in plain buffer for the heat denaturation of COMP coiled-coil domain and its mutant obtained by extrapolation to zero molar guanidinium chloride concentration are approximately 160 and 220 degrees C, respectively, which groups them among the most stable proteins.

Journal Article↗

PubMed2000

Entry of R5X4 and X4 human immunodeficiency virus type 1 strains is mediated by negatively charged and tyrosine residues in the amino-terminal domain and the second extracellular loop of CXCR4.

CXCR4 mediates the fusion and entry of X4 and R5X4 strains of human immunodeficiency virus type 1 (HIV-1). The residues involved in CXCR4 coreceptor function have not all yet been identified, but tyrosine and negatively charged residues in the amino-terminal domain of CCR5 were shown to be indispensable for gp120 binding and entry of R5 and R5X4 strains. We therefore evaluated the role of such residues in CXCR4 coreceptor function by replacing tyrosines (Y), aspartic acids (D), and glutamic acids (E) with alanines (A) and testing the ability of these mutants to mediate the entry of X4 and R5X4 HIV-1 isolates. Our results show that viral entry depends on YDE-rich clusters in both the amino-terminus and the second extracellular loop of CXCR4. Different viral isolates vary in their dependence on residues in one or the other domain. The determinants of CXCR4 coreceptor function are, therefore, more diffuse and isolate-dependent than those of CCR5.

Amino Acid Sequence↗

PubMed2000

The development of abnormal axon trajectories after rotation of one eye in Xenopus.

The targeting of isthmotectal axons in the Xenopus binocular pathway is guided by both activity-dependent cues and activity-independent cues. Abnormal visual activity induced by unilateral eye rotation overrides activity-independent cues and causes isthmotectal axons to arborize at new locations during a critical period of development that ends approximately 3 months postmetamorphosis (PM). Horseradish peroxidase staining of isthmotectal axons reveals that they normally run rostrocaudally in the tectum; in contrast, those axons in animals with early eye rotation have circuitous trajectories. In this paper, by studying the trajectories and branching patterns of isthmotectal axons at different times after eye rotation, we aimed to investigate when and how activity cues determine the projection pattern of isthmotectal axons. As suggested by electrophysiological recording, isthmotectal axons initially grow normally and make arbors according to activity-independent cues despite the presence of abnormal visual input. Our findings demonstrate that the development of abnormal trajectories starts by 2 weeks PM in response to eye rotation and is a protracted process. It begins in the tectal regions in which the initial connections of isthmotectal axons are first formed according to activity-independent cues. At transitional stages (5 and 10 weeks), axons with arbors at two different locations are observed, with locations corresponding to the old and new termination sites, respectively. Later, at 10 weeks of age, the fainter horseradish peroxidase staining in arbors at old termination sites suggests that the older arbors are undergoing withdrawal.

Animals↗

PubMed2000

Two cases of AML (M2) with a t(8;19)(q22;q13): a new cytogenetic variant.

"Simple" variants of the t(8;21) translocation involving chromosome 8 and a chromosome other than number 21 are rare. To our knowledge, only t(3;8)(q29;q22), t(8;11)(q22;q13), t(8;16)(q22;q24), t(8;20)(q22;p13), and t(8;22) have been reported in the literature. This paper describes for the first time two patients with acute myelogenous leukemia with a consistent t(8;19)(q22;q13) translocation. Their myelograms were compatible with the FAB-M2 subtype. The blasts from case 2 expressed CD34, CD33, CD13, and CD19. Karyotype analyses were performed on bone marrow cells using R- and G-banding at presentation. A t(8;19)(q22;q13) translocation was found in 28/30 metaphases for case 1 and in 23/25 metaphases for case 2. The latter case also had a deletion of chromosome 9, del(9)(q12q22) as an additional abnormality. Reverse transcriptase-polymerase chain reaction study revealed no AML1/ETO fusion transcript in case 2. Dual-color fluorescence in situ hybridization (FISH) assay using two probes (BAC92 and YAC412A4) convincingly demonstrated that the chromosomal material from 8q was translocated onto 19q rather than 19p in case 2. Thus, we consider t(8;19)(q22;q13) a true "simple" variant of t(8;21), and assume that a fusion gene resulting from the t(8;19) may contain the ETO gene located at 8q22 and an unknown partner gene from 19q13, which probably is a new transcription factor, whose molecular entity warrants further study.

Adolescent↗

PubMed2000

Stimulation of phosphatidylinositol-4-phosphate 5-kinase by Rho-kinase.

The serine/threonine kinase Rho-kinase was recently identified as a downstream effector of the small GTPase Rho, mediating effects of Rho on the actin cytoskeleton. Also phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)) has been implicated in the regulation of actin polymerization. As the synthesis of PI(4,5)P(2) has been suggested to be affected by Rho proteins, we investigated whether Rho-kinase is involved in the control of PI(4,5)P(2) levels. Overexpression of RhoA in HEK-293 cells increased phosphatidylinositol 4-phosphate (PI4P) 5-kinase activity and concomitantly enhanced cellular PI(4,5)P(2) levels, whereas overexpression of the Rho-inactivating C3 transferase decreased both PI4P 5-kinase activity and PI(4,5)P(2) levels. These effects of RhoA could be mimicked by overexpression of wild-type Rho-kinase and of the constitutively active catalytic domain of Rho-kinase, Rho-kinase-CAT. In contrast, a kinase-deficient mutant of Rho-kinase had no effect on PI4P 5-kinase activity. Importantly, the increase in PI4P 5-kinase activity and PI(4,5)P(2) levels by wild-type Rho-kinase, but not by Rho-kinase-CAT, was completely prevented by coexpression of C3 transferase, indicating that the effect of Rho-kinase was under the control of endogenous Rho. In cell lysates, addition of recombinant RhoA and Rho-kinase-CAT stimulated PI4P 5-kinase activity. Finally, the increase in PI(4,5)P(2) levels induced by both Rho-kinase-CAT and RhoA was reversed by the Rho-kinase inhibitor HA-1077. Our data suggest that Rho-kinase is involved in the Rho-controlled synthesis of PI(4,5)P(2) by PI4P 5-kinase.

Cell Line↗

PubMed2000

Phosphorylation and destabilization of human period I clock protein by human casein kinase I epsilon.

Period (PER), a central component of the circadian clock in Drosophila, undergoes daily oscillation in abundance and phosphorylation state. Here we report that human casein kinase I epsilon (hCKI epsilon) can phosphorylate human PER I (hPER I). Purified recombinant hCKI epsilon (but not a kinase negative mutant of hCKI epsilon, hCKI epsilon-K38R) phosphorylated hPER I in vitro. When co-transfected with wild-type hCKI epsilon, in 293T cells, hPER I showed a significant increase in phosphorylation as evidenced by a shift in molecular mass. Furthermore, phosphorylation of hPER I by hCKI epsilon caused a decrease in protein stability in hPER I. Whereas phosphorylated hPER I had a half-life of approximately 12 h, unphosphorylated hPER I remained stable in the cell for > 24 h. hPER I protein could also be co-immunoprecipitated with transfected hCKI epsilon as well as endogenous hCKI epsilon, indicating physical association between hPER I and hCKI epsilon proteins in vivo.

Animals↗

PubMed2000

Identification of caspase 3-mediated cleavage and functional alteration of eukaryotic initiation factor 2alpha in apoptosis.

Induction of apoptosis in a variety of cell types leads to inhibition of protein synthesis. Recently, the cleavage of eukaryotic translation initiation factor 4G (eIF4G) by caspase 3 was described as a possible event contributing to translation inhibition. Here, we report the cleavage of another initiation factor in apoptotic cells, eIF2alpha, that could contribute to regulation of translation during apoptosis. This cleavage event could be completely inhibited by pretreatment of HeLa cells with Z-VAD-fmk. In vitro analysis using purified eIF2 and purified caspases showed cleavage of eIF2alpha by caspase 3, 6, 8, and 10 but not 9. Caspase 3 most efficiently cleaved eIF2alpha and this could be inhibited by addition of Ac-DEVD-CHO in vitro. Comparison of cleavage of phosphorylated versus nonphosphorylated eIF2alpha revealed a modest preference of the caspases for the nonphosphorylated form. When eIF2. 2B complex was used as substrate, only caspase 3 was capable of eIF2alpha cleavage, which was not affected by phosphorylation of the alpha subunit. The eIF2.GDP binary complex was cleaved much less efficiently by caspase 3. Sequence analysis of the cleavage fragment suggested that the cleavage site is located in the C-terminal portion of the protein. Analysis showed that after caspase cleavage, exchange of GDP bound to eIF2 was very rapid and no longer dependent upon eIF2B. Furthermore, in vitro translation experiments indicated that cleavage of eIF2alpha results in functional alteration of the eIF2 complex, which no longer stimulated upstream AUG selection on a mRNA containing a viral internal ribosome entry site and was no longer capable of stimulating overall translation. In conclusion, we describe here the cleavage of a translation initiation factor, eIF2alpha that could contribute to inhibition or alteration of protein synthesis during the late stages of apoptosis.

Amino Acid Sequence↗

PubMed2000

Heme oxygenase-1 induction protects murine cortical astrocytes from hemoglobin toxicity.

Exposure to micromolar concentrations of hemoglobin (Hb) results in the oxidative death of cultured cortical neurons, but glia are resistant. The role of heme oxygenase-1 (HO-1) induction on this glial resistance was investigated. Within two hours of exposure to 5 microM Hb, immunoblotting demonstrated an increase in HO-1 in confluent glial cultures. Consistent with prior observations, 23-30 h Hb exposure had little or no effect on glial viability, as assessed by lactate dehydrogenase release. Concomitant treatment with the HO inhibitors tin protoporphyrin IX or the D-amino acid peptide rvnlrialry resulted in release of 40-71% of glial lactate dehydrogenase; protein synthesis inhibition with cycloheximide produced a similar effect. These results are consistent with the hypothesis that HO-1 induction protects cortical astrocytes from Hb toxicity.

Animals↗

PubMed2000

Secular variation of iron isotopes in north atlantic deep water

A high-precision iron isotope time series for a ferromanganese crust demonstrates that the iron isotope composition in North Atlantic Deep Water has changed substantially over the past 6 million years and that iron isotope variations in the crust are closely correlated to those of lead isotopes. The close correlation between the two isotope series indicates that the observed iron isotope variations predominantly reflect those of iron input from terrigenous sources and provides no evidence for biologically induced mass fractionation within North Atlantic Deep Water.

Journal Article↗