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Biomedical subjects

H Zou

Publications and source records attributed to H Zou.

At least 109 records · Page 6Linked to original sources
PubMed1997

Distinct roles of type I bone morphogenetic protein receptors in the formation and differentiation of cartilage.

The bone morphogenetic proteins (BMPs), TGF beta superfamily members, play diverse roles in embryogenesis, but how the BMPs exert their action is unclear and how different BMP receptors (BMPRs) contribute to this process is not known. Here we demonstrate that the two type I BMPRs, BMPR-IA and BMPR-IB, regulate distinct processes during chick limb development. BmpR-IB expression in the embryonic limb prefigures the future cartilage primordium, and its activity is necessary for the initial steps of chondrogenesis. During later chondrogenesis, BmpR-IA is specifically expressed in prehypertrophic chondrocytes. BMPR-IA regulates chondrocyte differentiation, serving as a downstream mediator of Indian Hedgehog (IHH) function in both a local signaling loop and a longer-range relay system to PTHrP. BMPR-IB also regulates apoptosis: Expression of activated BMPR-IB results in increased cell death, and we showed previously that dominant-negative BMPR-IB inhibits apoptosis. Our studies indicate that in TGF beta signaling systems, different type I receptor isoforms are dedicated to specific functions during embryogenesis.

Animals↗
PubMed1997

Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3.

We report here the purification and cDNA cloning of Apaf-1, a novel 130 kd protein from HeLa cell cytosol that participates in the cytochrome c-dependent activation of caspase-3. The NH2-terminal 85 amino acids of Apaf-1 show 21% identity and 53% similarity to the NH2-terminal prodomain of the Caenorhabditis elegans caspase, CED-3. This is followed by 320 amino acids that show 22% identity and 48% similarity to CED-4, a protein that is believed to initiate apoptosis in C. elegans. The COOH-terminal region of Apaf-1 comprises multiple WD repeats, which are proposed to mediate protein-protein interactions. Cytochrome c binds to Apaf-1, an event that may trigger the activation of caspase-3, leading to apoptosis.

Amino Acid Sequence↗
PubMed1997

Holliday junctions accumulate in replication mutants via a RecA homolog-independent mechanism.

The Holliday junction recombination intermediate, an X-shaped DNA molecule (xDNA), was analyzed at rDNA in mitotically growing yeast. In wild-type cells, xDNA is only detected at S phase, suggesting that recombination is stimulated to repair replication-related lesions. A search for mutations that increase the level of xDNA uncovered a gene encoding a subunit of DNA polymerase alpha. Systematic examination of replication mutants revealed that defects in polymerase alpha and delta but not the epsilon complex stimulate the level of xDNA. These xDNAs are Holliday junctions and not replication intermediates. The level of Holliday junctions is greatly reduced in rad52 mutants, but surprisingly, not in mutants defective in the three known mitotically expressed yeast RecA homologs.

DNA Polymerase II↗
PubMed1997

Glutathione S-transferase expression in hepatitis B virus-associated human hepatocellular carcinogenesis.

Hepatitis B virus (HBV) and aflatoxin B1 represent the main risk factors for the development of hepatocellular carcinoma (HCC) in areas endemic for liver cancer. The glutathione S-transferases (GSTs) are a family of Phase II detoxification enzymes that catalyze the conjugation of a wide variety of endogenous and exogenous toxins, including aflatoxin B1, with glutathione. This study characterizes the GST isoenzyme composition (alpha, mu, and pi) of both HBV-infected normal hepatic tissues and HCCs. Analysis of matched pairs of hepatic tissue (normal and tumor) from 32 HCC patients indicated that total GST activity was significantly higher in normal tissues than in tumor tissues, although the percentage of samples expressing GST alpha and pi was equivalent. GST mu was detected by Western blot in the normal tissue from 87.5% of the subjects possessing the GST M1 gene but only 28.6% of the corresponding tumor tissues. The GST activity of normal tissue from GST M1 null patients was significantly decreased as compared to that of subjects possessing the GST M1 gene (264.6 and 422.2 nmol/min/mg, respectively; P = 0.005). GST pi appeared to be overexpressed in the normal tissue of GST M1 null patients, a potential compensatory effect. Patients positive for HBV DNA had significantly lower GST activity than those who were HBV negative (302.1 versus 450.0 nmol/min/mg, respectively; P = 0.02). These results suggest that cellular protection within the human liver is compromised by HBV infection and further decreased during hepatocellular tumorigenesis.

Adult↗
PubMed1997

DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis.

We have identified and purified from HeLa cytosol a protein that induces DNA fragmentation in coincubated nuclei after it is activated by caspase-3. This protein, designated DNA Fragmentation Factor (DFF), is a heterodimer of 40 kDa and 45 kDa subunits. The amino acid sequence of the 45 kDa subunit, determined from its cDNA sequence, reveals it to be a novel protein. Caspase-3 cleaves the 45 kDa subunit at two sites to generate an active factor that produces DNA fragmentation without further requirement for caspase-3 or other cytosolic proteins. In cells undergoing apoptosis, the 45 kDa subunit is cleaved in the same pattern as it is cleaved by caspase-3 in vitro. These data delineate a direct signal transduction pathway during apoptosis: caspase-3 to DFF to DNA fragmentation.

Amino Acid Sequence↗
PubMed1997

An ATP-gated cation channel with some P2Z-like characteristics in gastric smooth muscle cells of toad.

1. Whole-cell and single-channel currents elicited by extracellular ATP were studied in freshly dissociated smooth muscle cells from the stomach of the toad Bufo marinus using standard patch clamp and microfluorimetric techniques. 2. This ATP-gated cation channel shares a number of pharmacological and functional properties with native rat myometrium receptors, certain native P2Z purinoceptors and the recently cloned P2X7 purinoceptor. But, unlike the last two, the ATP-gated channel does not mediate the formation of large non-specific pores. Thus, it may represent a novel member of the P2X or P2Z class. 3. Extracellular application of ATP (> or = 150 microM) elicited an inward whole-cell current at negative holding potentials that was inwardly rectifying and showed no sign of desensitization. Na+, Cs+ and, to a lesser degree, the organic cation choline served as charge carriers, but Cl- did not. Ratiometric fura-2 measurements indicated that the current is carried in part by Ca2+. The EC50 for ATP was 700 microM in solutions with a low divalent cation concentration. 4. ATP (> or = 100 microM) at the extracellular surface of cell-attached or excised patches elicited inwardly rectifying single-channel currents with a 22 pS conductance. Cl- did not serve as a charge carrier but both Na+ and Cs+ did, as did choline to a lesser extent. The mean open time of the channel was quite long, with a range in hundreds of milliseconds at a holding potential of -70 mV. 5. Mg2+ and Ca2+ decreased the magnitude of the ATP-induced whole-cell currents. Mg2+ decreased both the amplitude and the activity of ATP-activated single-channel currents. 6. ADP, UTP, P1, P5-di-adenosine pentaphosphate (AP5A), adenosine and alpha, beta-methylene ATP (alpha, beta-Me-ATP) did not induce significant whole-cell current. ATP-gamma-S and 2-methylthio ATP (2-Me-S-ATP) were significantly less effective than ATP in inducing whole-cell currents, whereas benzoylbenzoyl ATP (BzATP) was more effective. BzATP, alpha, beta-Me-ATP, ATP-gamma-S and 2-Me-S-ATP induced single-channel currents, but a higher concentration of alpha, beta-Me-ATP was required. 7. BzATP did not induce the formation of large non-specific pores, as assayed using mag-fura-2 as a high molecular mass probe.

Adenosine Triphosphate↗
PubMed1997

Studies on the precursors of strong mutagen [3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone]MX by chlorination of fractions from different waters.

The strong mutagen, [3-chloro-4-(dichloromethyl)-5-hydroxy- 2(5H)-furanone] MX, was found to be one of the most potent mutagens in drinking water. In this study, dissolved organic matters from river water and lake water were separated into several compound classes by sorbtion on a series of resin absorbents. After chlorine treatment of the fractions, MX was determined with GC/MS in the selected ion monitoring mode. Humic substances produced more MX on a TOC-basis than other fractions and contributed more to MX formation in the chlorinated natural waters. Some phenols were detected in the oxidation products of humic substances and therefore formation of MX may occur when some phenolic precursor structures in humic substances are treated with chlorine.

Absorption↗
PubMed1997

[Investigation on electrochemical behavior of emodin and its application].

Emodin showed a second-order derivative reduction wave with peak potential of -0.75 V (SCE) by single sweep oscillopolarography using H3BO3-Na2B4O7 (pH 8.50) as base solution. The peak height of emodin is proportional to the concentration in the range of 1.42 x 10(-7)-5.7 x 10(-6) mol.L-1 and 7.1 x 10(-6)-7.1 x 10(-5) mol.L-1, respectively. The detection limit is 0.7 x 10(-7) mol.L-1. It can be employed for the determination of emodin in Rheum officinale Baill with satisfaction. The electrochemical behavior of emodin was studied and the electrode reaction mechanism was proposed. In addition, it was found that emodin, aloe-emodin, chrysophanol, rhein and physcion can scavenge superoxide anion radical produced by the autoxidation of pyrogallol. The scavenging activity is in the order: emodin > aloe-emodin > chrysophanol > rhein > physcion.

Anthraquinones↗
PubMed1997

[Determination of aniline derivatives in waste water with reversed-phase high performance liquid chromatography].

A reversed-phase high performance liquid chromatographic method for the determination of aniline derivatives in waste-water is described. Chromatographic conditions were: column, Hypersil BDS (200 mm x 4.0 mm i.d., 5 microm); mobile phase, methanol-water, gradient elution; flow rate, 1.0 mL/min; detection wavelength, 254 nm. Quantification was carried out based on the external standard of aniline derivatives. The pH of waste water sample was adjusted to 11-12 with dilute NaOH solution prior to extraction with CH2Cl2. The extractives concentrated in K-D concentrator and filtered with FH 0.5 microm membrane. Finally make up the volume to 1 mL and test it with reversed-phase high performance liquid chromatography. The linearity range of calibration curve used was 0.005-0.3 microg. The minimum detectable quantities of aniline derivatives varied from 1.20 ng to 3.00 ng. The average recoveries were from 51.57% to 107.92%, and the RSDs of the method were from 1.88% to 5.98%. The method is simple, rapid, and accurate.

Aniline Compounds↗
PubMed1997

[Research on the variation of peak shape in capillary zone electrophoresis].

The influence of operation conditions, such as, injection time, applied voltage and effective length of capillary column, on parameters of peak shape in capillary zone electrophoresis (CZE) was investigated by using exponential-modified Gaussian (EMG) equation. The CE experiments were carried out by using the sample of derivatives of benzoic acid and small peptides. The experimens indicated that there was a significant effect of injection time on W1/2 and that the peaks changed from tailing to leading when the injection time was over 3 seconds in the case of pressure injection. It is recommended that an injection time less than 3 seconds can lead to a good column efficiency. The good linear relationships between tm and tau, sigma, W1/2 (t) with different separation voltages were observed. In the range of experimental conditions (11-27kV), higher voltage could lead to smaller zone dispersion, giving rapid and high efficient separation, and the voltage was insignificant on peak tailing. It was also shown that sigma, tau and W1/2 increased slightly with increasing the effective length of capillary column.

Electrophoresis, Capillary↗
PubMed1997

[Research on the reproducibility and selection of experimental conditions of capillary electrochromatography].

In this work, we prepared the packed columns for capillary electrochromatography (CEC) with 75microm and 100microm i.d., both of coupled mode and uncoupled mode. A mixture of thiourea and other three aromatic compounds was employed to evaluate the efficiency of the CEC columns. RSD values of retention times smaller than 2.12% was obtained on the home-made CEC system. The influence of pH value and the concentration of the organic solvent in the buffer on the retention behavior was investigated, giving a guide to the selection of experimental conditions. High speed analysis can be achieved by using high pH value buffers. Higher concentration of acetonitrile will improve the elution ability, while lower ocncentration of acetonitrile would improve the resolution. Seven kinds of neutral molecules were separated efficiently within 25 minutes. The results from the two modes of CEC columns were compared.

English Abstract↗
PubMed1997

[Determination of steroids in milk powder and birds' feces by high performance liquid chromatography coupled with evaporative light scattering detector].

In this paper, a new, simple and rapid method for the analysis of steroids by reversed-phase high performance liquid chromatography (HPLC) coupled with evaporative light scattering detector (ELSD) is presented. A good resolution of the 12 steroids is obtained within 35 min with a gradient elution using a binary mobile phase composed of methanol and water. The analytical column used was 2.0mm x 150mm, packed with BDS (particle size 5microm) and the mobile phase flow rate was 0.2mL/min. Nebulizer gas flow rate and drift tube temperature, the two critical parameters for determining the reproducibility and signal-to-noise ratio of ELSD were 2.00mL/min and 95 degrees C, respectively. The response of the ELSD was non-linear as a result of basic light-scattering principles, the calibration curve by plotting lnY vs lnX (Y, X stand for peak area and concentration, respectively) shows a good linearity in the range of 5-50mg/L with a correlation coefficient higher than 0.99 for individual steroid and with relative standard deviation less than 6.92%. This method has been applied to the determination of steroids in milk powder and birds' feces, solid phase extraction on reverse phase bonded C18 (5mL/500mg) was used to treat the samples efficiently. The recoveries of the steroids were between 84.0%-102.8%.

Animals↗
PubMed1997

Immobilized IDA-type Cu2+-chelating membrane affinity chromatography for purification of bovine liver catalase.

Cu2+-chelating membrane medium based on iminodiacetate-substituted modified short cotton cellulose was examined to purify bovine liver catalase (BLC) by affinity chromatography. The buffer pH notably affect BLC binding to the Cu2+-chelating membrane adsorbent. The results showed that under the chromatographic conditions chosen, 67.7% recovery of BLC attained with an overall 4.7-fold increase in specific activity in a single step. After purification of BLC, the chelating membrane adsorbent can easily be regenerated by imidazole or EDTA buffer. Different results were obtained by the two regeneration methods: the later is more effective than the former in reviving the purification characters of the chelating membrane adsorbent.

Animals↗
PubMed1996

Immunochromatographic analysis of bovine growth hormone releasing factor involving reversed-phase high-performance liquid chromatography-immunodetection.

We have developed high-performance immunoaffinity chromatography (HPIAC) methods for the detection and quantitation of bovine growth hormone releasing factor (GHRF), which could also be applicable to its metabolites in biofluids. These approaches have involved a combination of IAC using immobilized antibody (Ab) to GHRF, together with reversed-phase high-performance liquid chromatography (RP-HPLC) separations of initially isolated and concentrated protein, followed by selective detection, involving on-line immunodetection (ID) schemes. ID methods involved HPIAC supports of the Ab, together with synthesized Ab-fluorescein isothiocyanate conjugates. We have demonstrated optimization methods for each step of the entire hyphenated technique (IAC-HPLC-ID), and then actually quantitated GHRF using this overall system. The minimum detectable concentration was about 1 ng/5 ml (200 ppt) with fluorescence detection (excitation wavelength, 490 nm; emission wavelength, 510-650 nm). We have also tested a single blind, spiked biological sample (bovine plasma), spiked with a known level of GHRF. Accuracy (7.4%) and precision (S.D. = +/- 22%) were quite acceptable for a double immunoassay method.

Animals↗
PubMed1996

Requirement for BMP signaling in interdigital apoptosis and scale formation.

Interdigital cell death leads to regression of soft tissue between embryonic digits in many vertebrates. Although the signals that regulate interdigital apoptosis are not known, BMPs--signaling molecules of the transforming growth factor-beta superfamily--are expressed interdigitally. A dominant negative type I BMP receptor (dnBMPR-IB) was used here to block BMP signaling. Expression of dnBMPR in chicken embryonic hind limbs greatly reduced interdigital apoptosis and resulted in webbed feet. In addition, scales were transformed into feathers. The similarity of the webbing to webbed duck feet led to studies that indicate that BMPs are not expressed in the duck interdigit. These results indicate BMP signaling actively mediates cell death in the embryonic limb.

Animals↗
PubMed1996

Gene conversion plays the major role in controlling the stability of large tandem repeats in yeast.

The genomic stability of the rDNA tandem array in yeast is tightly controlled to allow sequence homogenization and at the same time prevent deleterious rearrangements. In our study, we show that gene conversion, and not unequal sister chromatid exchange, is the predominant recombination mechanism regulating the expansion and contraction of the rDNA array. Furthermore, we found that RAD52, which is essential for gene conversion, is required for marker duplication stimulated in the absence of the two yeast type I topoisomerases. Our results have implications for the mechanisms regulating genomic stability of repetitive sequence families found in all eukaryotes.

Base Sequence↗