Biomedical subjects

Y Zhao

Publications and source records attributed to Y Zhao.

At least 415 records · Page 23Linked to original sources

PubMed2000

[Study of polyaluminium ferric chloride (PAFC) aqueous solutions visible spectra: the relationship among their absorbance at 460 nm, basicities and total Fe(III) concentration of PAFC solutions].

The visible spectra of forty-five synthesized polyaluminium ferric chloride (PAFC) aqueous solutions with different [OH]/[Al + Fe] (B) and nAl/nFe(molar ratio) were investigated. The results showed that their absorbances A of shoulder at 460 nm are all linear interrelationship with their B. We made the curve of absorbance A of PAFC with same B vs. nAl/nFe, there was a turning point. All of these points just located at the places of pH value rising higher in pH- nAl/nFe diagrams. For these reason, we thought that these points stand for the dynamic equilibrium of OH- between the polynucleus hydroxyl Fe(III) and Al(III). These points made certain relationship among the absorbance A at 460 nm, basicities and total Fe(III) concentration of PAFC solutions.

English Abstract↗

PubMed2000

[Fluorescence studies on the mixed solid complexes of europium and terbium].

Complexes Eu2L3L' and Tb2L3L' (L = 0-phthalate, L' = 2,2'-dipyridyl) have been synthesized. Mixed the both complexes as different molar ratio, mechanically ground them, then obtained a series mixed solid complexes of europium and terbium. Their fluoresence and infrared spectra were investigated. The results showed that there were obviously change for their fluorescence intensities and peak positions as compared with Eu2L3L' or Tb2L3L'. The fluorescence intensities of europium ions were sensitized by terbium ions and those of terbium ions were quenched by europium ions. FTIR spectra indicated that there were a combined force between the two complexes in the mixed solid complexes.

Chelating Agents↗

PubMed2000

[Synthesis and characterization of complex of NdL3.2H2O].

The binary solid complexes using rare earth ion Nd3+ with the ligand L (L = O-chorophenoxyacetic acid and 3-methoxybenzoic acid) has been synthesized in this work. The chemical formula of the complex was determined to be NdL3.2H2O by elemental analysis. The composition structure and properties of this type of complex were tentatively investigated by the analysis of molar conductivity, TG-DTA, IR and UV.

2,4-Dichlorophenoxyacetic Acid↗

PubMed1999

A molecular basis for nitric oxide sensing by soluble guanylate cyclase.

Nitric oxide (NO) functions as a signaling agent by activation of the soluble isoform of guanylate cyclase (sGC), a heterodimeric hemoprotein. NO binds to the heme of sGC and triggers formation of cGMP from GTP. Here we report direct kinetic measurements of the multistep binding of NO to sGC and correlate these presteady state events with activation of enzyme catalysis. NO binds to sGC to form a six-coordinate, nonactivated, intermediate (k(on) > 1.4 x 10(8) M(-1).s(-1) at 4 degrees C). Subsequent release of the axial histidine heme ligand is shown to be the molecular step responsible for activation of the enzyme. The rate at which this step proceeds also depends on NO concentration (k = 2.4 x 10(5) M(-1).s(-1) at 4 degrees C), thus identifying a novel mode of regulation by NO. NO binding to the isolated heme domain of sGC was also rapid (k = 7.1 +/- 2 x 10(8) M(-1).s(-1) at 4 degrees C); however, no intermediate was observed. The data show that sGC acts as an extremely fast, specific, and highly efficient trap for NO and that cleavage of the iron-histidine bond provides the driving force for activation of sGC. In addition, the kinetic data indicate that transport or stabilization of NO is not necessary for effective signal transmission.

Flow Injection Analysis↗

PubMed1999

Isolated cleft palate in mice with a targeted mutation of the LIM homeobox gene lhx8.

Formation of the mammalian secondary palate is a highly regulated and complex process whose impairment often results in cleft palate, a common birth defect in both humans and animals. Loss-of-function analysis has linked a growing number of genes to this process. Here we report that Lhx8, a recently identified LIM homeobox gene, is expressed in the mesenchyme of the mouse palatal structures throughout their development. To test the function of Lhx8 in vivo, we generated a mutant mouse with a targeted deletion of the Lhx8 gene. Our analysis of the mutant animals revealed a crucial role for Lhx8 in palatogenesis. In Lhx8 homozygous mutant embryos, the bilateral primordial palatal shelves formed and elevated normally, but they often failed to make contact and to fuse properly, resulting in a cleft secondary palate. Because development of other craniofacial structures appeared normal, the impaired palatal formation in Lhx8-mutant mice was most likely caused by an intrinsic primary defect in the mesenchyme of the palatal shelves. The cleft palate phenotype observed in Lhx8-mutant mice suggests that Lhx8 is a candidate gene for the isolated nonsyndromic form of cleft palate in humans.

Animals↗

PubMed1999

Cytochrome c release and caspase activation during menadione-induced apoptosis in plants.

We report here the detection of the release of cytochrome c from mitochondria into the cytosol during menadione-induced apoptosis in tobacco protoplasts. Western blot analysis indicated that the caspase specific inhibitors AC-DEVD-CHO (Ac-Asp-Glu-Val-Asp-aldehyde) and AC-YVAD-CHO (N-acetyl-Try-Val-Ala-aspartinal) inhibited the degradation of a caspase 3 specific substrate PARP (poly(ADP-ribose) polymerase), and they had no effect on the release of cytochrome c. Further study showed that menadione could not induce apoptosis of mouse liver nuclei in tobacco cytosol extract containing no mitochondria. However, when cytochrome c or mitochondria was added into the cytosol extract, apoptosis of mouse liver nuclei and the degradation of PARP could both be detected. The results provide strong evidence that menadione can induce apoptosis in tobacco protoplasts via the release of cytochrome c from mitochondria into the cytosol.

Animals↗

PubMed1999

Donor-type chimerism determination by competitive polymerase chain reaction (PCR) in a primate model for bone marrow transplantation.

BACKGROUND: Evaluation of the outcome of successful bone marrow transplantation (BMT) and in-depth studies of transplantation biology rely increasingly on accurate detection of donor origin cells in the transplanted recipients. This study describes a quantitative competitive polymerase chain reaction (PCR) assay for accurate evaluation of chimerism after allogeneic BMT in a cynomologous primate model, based on detection of monkey Y-specific DNA. METHODS: A competitor standard was generated via PCR using a mutagenic primer that makes the competitor DNA 22 bp less than the wild type monkey Y-specific DNA. The mutated form can still be amplified by the primer pair for the detection of monkey Y-specific DNA. A fixed amount of sample subjected to chimerism detection was co-amplified with a range of competitor DNA using a touch down program and hot start PCR technique. The PCR products were analyzed by computing densitometry. The ratio of competitor/target (Y-specific) DNA for each sample pair was calculated. RESULTS: Using DNAs prepared from an artificial mixture of male and female cells, a set of standard curves has been obtained and the sensitivity of the established quantitative PCR was found to be 25 pg of male DNA, which corresponds approximately to 0.005 fg competitor DNA. A DNA sample taken from a female monkey, transplanted with purified CD34+ stem cells from a male monkey donor 26 days after BMT, was subjected to the competitive PCR with 10% male DNA as a control; the level of male DNA in this sample was calculated to be around 50%. CONCLUSIONS: This quantitative PCR assay offers both a high degree of specificity as well as a very accurate and sensitive evaluation of chimerism in a sex-mismatched monkey BMT model.

Animals↗

PubMed1999

Cloning and mapping of a novel human serum/glucocorticoid regulated kinase-like gene, SGKL, to chromosome 8q12.3-q13.1.

Serum/glucocorticoid regulated kinase (sgk) belongs to a newly emerging subfamily of the serine/threonine protein kinase family. Although human SGK shares 98% amino acid identity with rat sgk, their expression levels are regulated differently, which indicates the existence of other SGKs in humans. In this paper, we reported the cloning of human SGKL, which encodes a protein sharing 67 and 66% amino acid identity with rat sgk and human SGK, respectively. A 4.4-kb transcript of human SGKL was detected in 16 human tissues examined and was found to be most abundant in lung. By radiation hybrid mapping, the SGKL gene was located to human chromosome 8q12. 1-q13.1 between markers D8S510 and D8S1797.

Adrenocorticotropic Hormone↗

PubMed1999

Crystal structure of an archaebacterial DNA polymerase.

BACKGROUND: Members of the Pol II family of DNA polymerases are responsible for chromosomal replication in eukaryotes, and carry out highly processive DNA replication when attached to ring-shaped processivity clamps. The sequences of Pol II polymerases are distinct from those of members of the well-studied Pol I family of DNA polymerases. The DNA polymerase from the archaebacterium Desulfurococcus strain Tok (D. Tok Pol) is a member of the Pol II family that retains catalytic activity at elevated temperatures. RESULTS: The crystal structure of D. Tok Pol has been determined at 2.4 A resolution. The architecture of this Pol II type DNA polymerase resembles that of the DNA polymerase from the bacteriophage RB69, with which it shares less than approximately 20% sequence identity. As in RB69, the central catalytic region of the DNA polymerase is located within the 'palm' subdomain and is strikingly similar in structure to the corresponding regions of Pol I type DNA polymerases. The structural scaffold that surrounds the catalytic core in D. Tok Pol is unrelated in structure to that of Pol I type polymerases. The 3'-5' proofreading exonuclease domain of D. Tok Pol resembles the corresponding domains of RB69 Pol and Pol I type DNA polymerases. The exonuclease domain in D. Tok Pol is located in the same position relative to the polymerase domain as seen in RB69, and on the opposite side of the palm subdomain compared to its location in Pol I type polymerases. The N-terminal domain of D. Tok Pol has structural similarity to RNA-binding domains. Sequence alignments suggest that this domain is conserved in the eukaryotic DNA polymerases delta and epsilon. CONCLUSIONS: The structure of D. Tok Pol confirms that the modes of binding of the template and extrusion of newly synthesized duplex DNA are likely to be similar in both Pol II and Pol I type DNA polymerases. However, the mechanism by which the newly synthesized product transits in and out of the proofreading exonuclease domain has to be quite different. The discovery of a domain that seems to be an RNA-binding module raises the possibility that Pol II family members interact with RNA.

Amino Acid Sequence↗

PubMed1999

Tenascin is expressed in the mesenchyme of the embryonic lung and down-regulated by dexamethasone in early organogenesis.

Tenascin (TN) is a hexameric extracellular matrix glycoprotein that is temporally and spatially restricted during lung development. This study examines the expression and regulation of TN in early lung organogenesis. Two TN isoforms were detected in total RNA isolated from embryonic day 14 rat lung tissues by reverse transcriptase polymerase chain reaction. The localization of TN in embryonic day 14 rat lung tissues was investigated by using in situ hybridization performed with an antisense RNA probe. TN mRNA was expressed exclusively by the mesenchyme but not by the epithelium of embryonic rat lungs. The intense expression of TN was observed in the mesenchyme that immediately surrounds the growing epithelial cells of the developing bronchi. The effect of the synthetic glucocorticoid hormone dexamethasone on the regulation of TN expression was examined by in vitro lung explant culture. Two TN polypeptides, the larger (M(r) 230 kDa, TN230) polypeptide and the smaller (M(r) 180 kDa, TN180) isoform, were detected in embryonic day 21 rat lungs by immunoblot analysis with anti-TN antibody. Dexamethasone inhibited both TN230 and TN180 biosynthesis. The study demonstrates the expression of TN at the early stage of lung organogenesis and presents evidence of hormonal regulation of TN in lung development, suggesting a potential role of TN in the communication between the epithelial and mesenchymal cells during lung branching morphogenesis.

Animals↗

PubMed1999

Combinatorial search of substituted beta-cyclodextrins for phosphatase-like activity.

Thirteen per-6-akylamino-6-deoxy-beta-cyclodextrin libraries (beta-CD libraries) were generated by a solution-phase combinatorial synthesis starting from per-6-iodo-6-deoxy-beta-CD and different combinations of eleven individual amine nucleophiles. Certain libraries showed the ability to hydrolyzep-nitrophenyl phosphate in the presence of Zn2+.

Combinatorial Chemistry Techniques↗

PubMed1999

pH-mediated field amplification on-column preconcentration of anions in physiological samples for capillary electrophoresis.

Two limitations of capillary electrophoresis (CE) are the low sample loadability of the capillary and an incompatibility with high ionic strength samples. Several strategies have been described to preconcentrate and lower the ionic strength of physiological samples prior to CE analysis. These have included both off-capillary and on-capillary approaches. We have previously described a version of on-column field-amplification stacking termed pH-mediated stacking. pH-mediated stacking was initially developed for the separation of cations. In this report, we describe the application of pH-mediated sample stacking to anions. In this method, an electrokinetic injection is used to introduce analyte anions into the CE system and simultaneously replace the sample matrix cations with ammonia from the background electrolyte. Base is then electrokinetically injected to neutralize the sample zone and create a low conductivity region across which the analyte anions will stack. Using this method, a sensitivity enhancement of more than 66-fold was achieved without loss in separation efficiency relative to normal electrokinetic injection. Detection limits of 0.3 microM for four phenolic acids in a physiological sample were achieved using simple UV absorbance detection. The limit to the amount of sample that could be loaded using this technique was the length of the separation capillary. To further increase the amount of sample that could be loaded, a double-capillary system was developed. Using the double-capillary system the sensitivity was increased more than 300-fold and detection limits of 0.06 microM were achieved.

Ammonia↗

PubMed1999

Quantification of cellular properties from external fields and resulting induced velocity: cellular hydrodynamic diameter.

An experimental technique is discussed in which the size distribution of a population of cells is determined by calculating each cell's settling velocity. The settling velocity is determined from microscopically obtained images which were recorded on SVHS tape. These images are then computer imaged and processed, and the cell's location and velocity are determined using a computer algorithm referred to as cell tracking velocimetry (CTV). Experimental data is presented comparing the distribution of human lymphocytes and a human breast cancer cell line, MCF-7, determined using a Coulter counter and the CTV approach.

Algorithms↗

PubMed1999

Quantification of cellular properties from external fields and resulting induced velocity: magnetic susceptibility.

An experimental technique is discussed in which the magnetic susceptibility of immunomagnetically labeled cells can be determined on a cell-by-cell basis. This technique is based on determining the magnetically induced velocity that an immunomagnetically labeled cell has in a well-defined magnetic energy gradient. This velocity is determined through the use of video recordings of microscopic images of cells moving in the magnetic energy gradient. These video images are then computer digitized and processed using a computer algorithm, cell tracking velocimetry, which allows larger numbers (>10(3)) of cells to be analyzed.

Cells, Cultured↗

PubMed1999

In situ fiber-optic oxygen consumption measurements from a working mouse heart.

Luminescence-based imaging-fiber oxygen sensors (IFOSs) were utilized for the in situ measurement of oxygen consumption from intact perfused mouse hearts. IFOSs were fabricated using a technically expedient, photoinitiated polymerization reaction whereby an oxygen-sensitive polymer matrix was immobilized in a precise location on an imaging fiber's distal face. The oxygen-sensing layer used in this work comprised a transition metal complex, Ru(Ph2phen)3(2+), entrapped in a gaspermeable photopolymerizable siloxane membrane (PS802). The transduction mechanism was based upon the oxygen collisional quenching of the ruthenium complex luminescence; detection was performed utilizing an epi-fluorescence microscope/charge coupled device imaging system. IFOS measurements from working mouse hearts were validated through concurrent, blind, ex situ blood gas analyzer (BGA) measurements. The BGA and IFOS methodologies were utilized successfully to measure oxygen concentrations in aortic and pulmonary artery perfusates from the working mouse heart before and after isoproterenol administration. Coupled with coronary-flow measurements, these data were used to calculate myocardial oxygen consumption. Regression analysis of measurements of myocardial oxygen consumption showed that there was a strong correlation between the values generated by the BGA sampling and those obtained via in situ IFOS methods. To our knowledge, this research represents the first report of in situ fiber-optic sensor monitoring of oxygen content from the intact, beating mouse heart.

Animals↗

PubMed1999

Bid-deficient mice are resistant to Fas-induced hepatocellular apoptosis.

The protein Bid is a participant in the pathway that leads to cell death (apoptosis), mediating the release of cytochrome c from mitochondria in response to signals from 'death' receptors known as TNFR1/Fas on the cell surface. It is a member of the proapoptotic Bcd-2 family and is activated as a result of its cleavage by caspase 8, one of a family of proteolytic cell-death proteins. To investigate the role of Bid in vivo, we have generated mice deficient for Bid. We find that when these mice are injected with an antibody directed against Fas, they nearly all survive, whereas wild-type mice die from hepatocellular apoptosis and haemorrhagic necrosis. About half of the Bid-deficient animals had no apparent liver injury and showed no evidence of activation of the effector caspases 3 and 7, although the initiator caspase 8 had been activated. Other Bid-deficient mice survived with only moderate damage: all three caspases (8 and 37) were activated but their cell nuclei were intact and no mitochondrial cytochrome c was released. We also investigated the effects of Bid deficiency in cultured cells treated with anti-Fas antibody (hepatocytes and thymocytes) or with TNFalpha. (fibroblasts). In these Bid-/- cells, mitochondrial dysfunction was delayed, cytochrome c was not released, effector caspase activity was reduced and the cleavage of apoptosis substrates was altered. This loss-of-function model indicates that Bid is a critical substrate in vivo for signalling by death-receptor agonists, which mediates a mitochondrial amplification loop that is essential for the apoptosis of selected cells.

Animals↗

PubMed1999

Transmembrane distribution of substrate and product during the bioreduction of acetophenone with resting cells of Saccharomyces cerevisiae.

The average volumetric intracellular concentrations of acetophenone and phenethyl alcohol were determined during the bioreduction of acetophenone using resting cells of Saccharomyces cerevisiae in aqueous solutions at 30 degrees C. The behavior of their distribution coefficients (ratio of intracellular to extracellular concentrations) during the bioreduction process was evaluated with different cell preparation and extracellular conditions. The distribution coefficient of acetophenone was found to be in the range of 2.3-4.0. The distribution coefficient of phenethyl alcohol was found to be in the range of 1.3-1.8. Both the distribution coefficients were correlated significantly only with the physiological state of the resting cells as reflected by the relative cell mass (0.65-1.09). The correlation is approximately linear with the largest slope for the toxic reagent, acetophenone. No significant effects on the distribution coefficients were experimentally observed or were present in a regression analysis for the concentrations of acetophenone (0-0.30% v/v), phenethyl alcohol (0-0.20% v/v), ethanol (1.60-2.25% v/v), the extracellular pH (pH 2-7), or the presence of the salts: KCl, KH2PO4, MgSO4, NaCl, and CaCl2 (each 0-0.1 M) in the medium. Different cell initialization times (0-6 days) and initialization conditions were also included.

Acetophenones↗

PubMed1999

A predictive thermodynamic model for the bioreduction of acetophenone to phenethyl alcohol using resting cells of Saccharomyces cerevisiae.

Equilibrium conversions were observed in the range of 60.2-76.0% with different initial compositions of reaction media for the bioreduction of acetophenone using resting cells of Saccharomyces cerevisiae in aqueous solutions at 30 degrees C. The reduction of acetophenone in the cells under anaerobic conditions is considered to be coupled with the oxidation of ethanol to acetate in the cytoplasm. A biphasic thermodynamic model is proposed which includes a nonuniform distribution of reagents across the cell membrane, a transmembrane pH gradient, ideal and nonideal solution models, and a basic reaction stoichiometry (ACP + (1/2) EtOH + (1/2)H2O <--> PEA + (1/2)Ac- + (1/2)H+). The intracellular activity coefficients were based on the Lewis-Randall rule for acetophenone, phenethyl alcohol, and H2O and Henry's law for ethanol, acetate anion, and H+. The overall standard Gibbs free energy was estimated to be -0.11 kcal/mol at a pH 7, 25 degrees C, and 1 atm. The intracellular thermodynamic activity coefficients of acetophenone and phenethyl alcohol were predicted to be 471.2 and 866.4, respectively, using the measured initial distribution coefficients and calculated extracellular activity coefficients. The model reflected a zero Gibbs free energy change at calculated conversions within 4% of the measured equilibrium conversions. The analysis verified the effect of the concentration ratio of the substrate acetophenone to the co-substrate ethanol on the conversion efficiency and suggested that the intracellular pH and the pH gradient across the cell transmembrane significantly affect the predicted equilibrium conversion. The intracellular pH of resting, viable cells of Bakers' yeast at the bioconversion conditions was determined experimentally to be 5.77.

Acetophenones↗