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Biomedical subjects

X Zou

Publications and source records attributed to X Zou.

At least 55 records · Page 3Linked to original sources

B-cell tumorigenesis in mice carrying a yeast artificial chromosome-based immunoglobulin heavy/c-myc translocus is independent of the heavy chain intron enhancer (Emu).

We have used YAC (yeast artificial chromosome) technology to create large translocation regions where the c-myc proto-oncogene is coupled to the core region of the human immunoglobulin heavy chain (IgH) locus (from VH2-5 through to Cdelta). Chimeric mice were obtained from embryonic stem cells carrying a single copy of the 240-kb IgH/c-myc translocation region. B-cell tumorigenesis occurs in the translocus mice, even when the entire Emu intron enhancer region between the joining segments and switch mu is deleted. This demonstrates that as yet unidentified regulatory elements in the IgH locus, independent from the known enhancers, are sufficient to cause B-cell specific activation of c-myc after translocation. The phenotype of tumors from IgH/c-myc YAC transgenic mice with or without Emu (B220+, IgM+/IgD+) is reminiscent of Burkitt's lymphoma. A rapidly expanding abnormal B-cell population is present at birth and accumulates in bone marrow, periphery, and spleen, well before discrete tumor establishment. Molecular analysis identified a clonal origin, with rearrangement of one mouse heavy chain allele retained in tumor cells from different sites, whereas subsequent rearrangements of heavy or light chain loci can be diverse. These mice routinely develop mature B-cell tumors early in life and may provide an invaluable resource of a B-cell lymphoma model.

Age Factors↗

Measurement of crystal thickness and crystal tilt from HRTEM images and a way to correct for their effects.

The effects of thickness and tilt angle are studied numerically on experimental high-resolution transmission electron microscope (HRTEM) images of a wedge-shaped metal oxide crystal. For sufficiently thin and well-aligned crystals, the amplitudes and phases of the Fourier transforms of the HRTEM images are essentially the same as the crystallographic structure factors. For tilted crystals, the changes of amplitudes and phases as a function of increased thickness and tilt angle can be described by a simple model. A method is presented by which the local thickness can be determined from one HRTEM image and one convergent-beam electron diffraction pattern from the same crystal. It is also shown how the projected potential can be reconstructed from HRTEM images of tilted crystals, disclosing the crystal structure, even from quite thick (>20 nm) samples.

Crystallography↗

On the phase problem in electron microscopy: the relationship between structure factors, exit waves, and HREM images.

In electron microscopy, the word phase is used for different physical phenomena, including crystallographic structure-factor phases and the electron wave phases. This has resulted in great confusion, as to whether the phase information is present or lost when an image is recorded. The aim of this paper is to solve this phase confusion problem by studying the relationships between structure factors, exit waves, and high-resolution electron microscopy (HREM) images. Three approaches are taken. First phases at different stages of the imaging processes are compared analytically for a crystal that can be considered a weak phase object (WPO). Then these different phases are calculated by the multi-slice method based on dynamical diffraction theory, and their numerical values are compared. Finally, the validity of the theoretical description is checked by comparison with experimental data on a real crystal, Ti(2)S. It is demonstrated that it is possible to obtain accurate structure factor-phases directly from HREM images by crystallographic image processing. The two major methods for structure determination from HREM images-exit wave reconstruction and crystallographic image processing-are compared. It is shown that the information utilised by the two methods as well as the results are essentially the same.

Crystallography↗

A human immunoglobulin lambda locus is similarly well expressed in mice and humans.

Transgenic mice carrying a 380-kb region of the human immunoglobulin (Ig) lambda light (L) chain locus in germline configuration were created. The introduced translocus on a yeast artificial chromosome (YAC) accommodates the most proximal Iglambda variable region (V) gene cluster, including 15 Vlambda genes that contribute to >60% of lambda L chains in humans, all Jlambda-Clambda segments, and the 3' enhancer. HuIglambdaYAC mice were bred with animals in which mouse Igkappa production was silenced by gene targeting. In the kappa-/- background, human Iglambda was expressed by approximately 84% of splenic B cells. A striking result was that human Iglambda was also produced at high levels in mice with normal kappa locus. Analysis of bone marrow cells showed that human Iglambda and mouse Igkappa were expressed at similar levels throughout B cell development, suggesting that the Iglambda translocus and the endogenous kappa locus rearrange independently and with equal efficiency at the same developmental stage. This is further supported by the finding that in hybridomas expressing human Iglambda the endogenous L chain loci were in germline configuration. The presence of somatic hypermutation in the human Vlambda genes indicated that the Iglambda-expressing cells function normally. The finding that human lambda genes can be utilized with similar efficiency in mice and humans implies that L chain expression is critically dependent on the configuration of the locus.

Animals↗

Stable isotopic studies of earthworm feeding ecology in tropical ecosystems of Puerto Rico.

Feeding strategies of earthworms and their influence on soil processes are often inferred from morphological, behavioral and physiological traits. We used (13)C and (15)N natural abundance in earthworms, soils and plants to explore patterns of resource utilization by different species of earthworms in three tropical ecosystems in Puerto Rico. In a high altitude dwarf forest, native earthworms Trigaster longissimus and Estherella sp. showed less (15)N enrichment ((15)N = 3-6 per thousand) than exotic Pontoscolex corethrurus ((15)N =7-9 per thousand) indicating different food sources or stronger isotopic discrimination by the latter. Conversely, in a lower altitude tabonuco forest, Estherella sp. and P. corethrurus overlapped completely in (15)N enrichment ((15)N = 6-9 per thousand), suggesting the potential for interspecific competition for N resources. A tabonuco forest converted to pasture contained only P. corethrurus which were less enriched in (15)N than those in the forest sites, but more highly enriched in (13)C suggesting assimilation of C from the predominant C(4) grass. These results support the utility of stable isotopes to delineate resource partitioning and potential competitive interactions among earthworm species. Copyright 1999 John Wiley & Sons, Ltd.

Journal Article↗

Monitoring catalytic reaction of bilirubin oxidase and determination of bilirubin and bilirubin oxidase activity by capillary electrophoresis.

Micellar electrokinetic chromatography (MEKC) has been exploited as an analytical method for the separation as well as for the determination of serum bilirubin fractions (alpha-, beta-, gamma- and delta-fraction), the activity of bilirubin oxidase (BOD), and for monitoring the catalytic reaction of BOD. The samples were followed by detection at 450 nm. The average limit of detection was in the 5.0 microM concentration range. The relative standard deviation (RSD) of migration times and peak area of bilirubin was less than 0.25% and 3.0%, respectively. This allowed us complete separation of the four bilirubin fractions (alpha-, beta-, gamma- and delta-fraction) and monitoring of the oxidation progress and the assay of the activity of bilirubin oxidase. The MEKC method is a convenient technique for studying the catalytic reaction of bilirubin oxidase at different pH and assaying the relationship between the four bilirubin fractions and diseases.

Bilirubin↗

Interaction of maturation delay and nonlinear birth in population and epidemic models.

A population with birth rate function B(N) N and linear death rate for the adult stage is assumed to have a maturation delay T>0. Thus the growth equation N'(t)=B(N(t-T)) N(t-T) e(-)d(1)T- dN(t) governs the adult population, with the death rate in previous life stages d(1)>==0. Standard assumptions are made on B(N) so that a unique equilibrium N(e) exists. When B(N) N is not monotone, the delay T can qualitatively change the dynamics. For some fixed values of the parameters with d(1)>0, as T increases the equilibrium N(e) can switch from being stable to unstable (with numerically observed periodic solutions) and then back to stable. When disease that does not cause death is introduced into the population, a threshold parameter R(0) is identified. When R(0)<1, the disease dies out; when R(0)>1, the disease remains endemic, either tending to an equilibrium value or oscillating about this value. Numerical simulations indicate that oscillations can also be induced by disease related death in a model with maturation delay.

Adult↗

Postprandial changes of sympathovagal balance measured by heart rate variability.

The aim of this study was to investigate the sympathovagal balance after meals by measuring the spectral analysis of heart rate variability (HRV). Nine healthy volunteers were enrolled in this study. The electrocardiogram (ECG) was recorded for 30 min in a fasting state and 60 min after a 500-kcal test meal. The HRV was derived from the ECG and was measured by power spectral analysis using fast-Fourier transform algorithm. It reveals two dominant spectral components. The low-frequency (LF) band reflects primarily sympathetic activity with some parasympathetic input. The high-frequency (HF) band is a reflection of parasympathetic (vagal) activity. The LF-to-HF ratio is considered a marker of sympathovagal balance. It was found that the postprandial LF-to-HF ratio, compared with the fasting state, was significantly increased at both the first 30 min (2.50 +/- 0.49 vs 1.78 +/- 0.33, P < 0.05) and the second 30 min (2.68 +/- 0.55 vs 1.78 +/- 0.33, P < 0.05). The postprandial HF diminished significantly at both the first (16.0 +/- 0.5 vs 21.8 +/- 4.2, P < 0.05) and the second (13.8 +/- 9.5 vs 21.8 +/- 4.2, P < 0.05) 30-min period. In conclusion, the postprandial sympathovagal ratio shows a sustained elevation lasting 1 hr, mainly attributed to diminished vagal activity.

Adult↗

Detection of gastric slow wave propagation from the cutaneous electrogastrogram.

The gastric slow wave is originated in the proximal stomach and propagates distally toward the pylorus. It determines the maximum frequency and propagation of gastric contractions. The aim of this study was to detect the propagation of the gastric slow wave from the surface electrogastrogram (EGG). The study was performed in 11 healthy subjects of normal weight. Gastric myoelectrical activity was recorded for 1 h in the fasting state with the use of a specially designed multichannel recording device that was composed of four identical amplifiers with cutoff frequencies of 1.8 and 16.0 cycles/min. Four active electrodes were placed on the abdomen along the gastric axis and were connected to a common reference electrode placed near the xiphoid process, yielding four-channel bipolar EGG signals. Cross-covariance analysis was performed to compute the time lag among the four channels. There was a time lag in EGG waveform between channels 1 and 4 (9.6 +/- 1.1 s); the average time during which the time lag was observed (>/=3 s) was 89.9 +/- 9.0%. There was a significant difference in the time lag among different adjacent channels (P < 0.04); the time lag observed between channels 3 and 4 was significantly smaller than that between channels 1 and 2 (P < 0.03). No correlation was found between the body mass index and the time lag between channels 1 and 4 (r = -0.31, P = 0.3). It was concluded that, with a multichannel recording device with identical multiamplifiers and an appropriate arrangement of abdominal electrodes, the propagation of the gastric slow wave could be identified from the EGG in healthy subjects. This method may be used to detect the coupling of the gastric slow wave noninvasively.

Adult↗

Induction of inflammatory mediators in human airway epithelial cells by lipid ozonation products.

We have proposed that exposure of epithelial cell membrane lipids in the lung (mainly phospholipids) to ozone will generate lipid ozonation products (LOP), which could be responsible for the proinflammatory effects of ozone. The ozonation of phosphocholine, the principal membrane phospholipid, produces a limited number of LOP, including hydroxyhydroperoxides and aldehydes. We now report that exposure of cultured human bronchial epithelial cells to the ozonized 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) product, 1-palmitoyl-2-(9-oxononanoyl)-sn-glycero-3-phosphocholine (PC-ALD), a phospholipase A(2) (PLA(2))-stimulatory LOP, resulted in a 113 +/- 11% increase in the amounts of tritiated platelet-activating factor ((3)H-PAF) released apically. (3)H-PAF release was also induced by 1-hydroxy-1-hydroperoxynonane of ozonized POPC (HHP-C9), a phospholipase C (PLC)- stimulatory LOP (134 +/- 40% increase in (3)H-PAF). PC-ALD at 10 microM, but not HHP-C9, induced a 127 +/- 24% increase in prostaglandin E(2) (PGE(2)) release (n = 6, p < 0.05). In contrast, HHP-C9, but not PC-ALD, induced interleukin (IL)-6 release (178 +/- 23% increase, n = 6, p < 0.05) and IL-8 release (101 +/- 23% increase, n = 8, p < 0. 05). These results suggest that LOP-dependent release of proinflammatory mediators may play an important role in the early inflammatory response seen during exposure to ozone.

Alkanes↗

[Investigation of factors affecting the development of COPD].

OBJECTIVE: To investigate the factors which affect the development of COPD. METHODS: 88 patients with mPAP less than 20 mm Hg were followed up by 5 - 20 years. Pulmonary ventilation, blood gas analysis, ECG, pulmonary hemodynamic parameters and oxygen saturation at night were performed one time every 5 years. RESULTS: The results showed that: 5 year, 10 year and 20 year's mortality is 6%, 25% and 43% respectively; MVV%, FEV(1)% and PaO(2) declined accompanied by the increase of PaCO(2); among 41 patients with mPAP 16 - 20 mm Hg, 20 patients' mPAP were more than 20 mm Hg and 15 patients died after 10 years' follow up. Among 16 patients with the declination of oxygen saturation, 8 patients' mPAP were more than 20 mm Hg and 6 patients died. 57 strains of infectious bacteria were found in 88 patients' sputum with 43 strains in the death group. CONCLUSIONS: These results demonstrated that COPD is a kind of chronic diseases with high death rate; infection, decline of oxygen saturation at night and mPAP more than 16 mm Hg were high risk factors in COPD.

Adult↗

[Magnetic resonance imaging in 40 cases of optic neuritis].

OBJECTIVE: To explore the clinical value of magnetic resonance imaging (MRI) in optic neuritis. METHODS: Forty patients with optic neuritis were examined with the spectral presaturation inversion recovery and fluid attenuated inversion recovery. RESULTS: The lesion was frequently seen in orbital portion (86.7%) and intracanalicular portion (29.3%), and primarily in the axial part (49.3%) and transverse section (38.7%) of the optic nerve. The longitudinal extent of lesion was from 2 to 35 mm (mean, 12.6 mm), the visual acuity of patients with optic neuritis was related to the site of the lesion, and negatively correlated with the longitudinal extent of lesion of logarithm curve. A poor visual acuity (< 0.01) was significantly associated with lesion of greater extent > 10 mm and the transverse or axial lesion, but the good visual acuity (>or= 0.3), with the lesion of smaller extent < 10 mm and the axial or peripheral lesion. In addition, paramagnetic contrast-medium, Gd-DTPA, enhancement scanning was carried out in 11 cases of optic neuritis, of them the visual acuity was increased in 10 cases after administration of corticosteroid, and in one case, visual improvement was not seen. CONCLUSION: MRI is a new and useful equipment which is helpful in the diagnosis and treatment of optic neuritis.

Adolescent↗

[Distribution of nitric oxide synthase-positive neurons in the hypothalamus of cat].

NADPH-diaphorase histochemistry was used to study the distribution of Nitric Oxide Synthase (NOS)-Positive neurons in the hypothalamus of adult cat. NOS-positive neurons were observed in the paraventricular hypothalamic nucleus, suprachiasmatic nucleus, dorsomedial hypothalamic nucleus, perifornical nucleus, anterior hypothalamic area and lateral hypothalamic area. Two differences between the cat and rat were noticed. Firstly, NOS-positive neurons were observed in the suprachiasmatic nucleus of cat, but the neurons in that of rat were negative for NOS. Secondly, the neurons in supraoptic nucleus of rat exhibited an intensive reaction to NADPH-diaphorase, but those in that of cat were faintly visible.

Animals↗

Structural and enzymatic studies of a new analogue of coenzyme B12 with an alpha-adenosyl upper axial ligand.

A new analogue of coenzyme B12 (5'-deoxyadenosylcobalamin, AdoCbl), in which the configuration of the N-glycosidic bond in the Ado ligand is inverted [(alpha-ribo)AdoCbl], has been synthesized and its crystal structure determined by X-ray diffraction [MoKalpha, lambda = 0.71073 A, monoclinic P212121, a = 16.132(12) A, b = 21. 684(15) A, c = 27.30(3) A, 9611 independent reflections, R1 = 0. 0708]. As suggested by molecular mechanics modeling before the structure was known, the Ado ligand lies over the southern quadrant of the molecule, as is the case for AdoCbl. The most striking feature of the structure is disorder in the orientation of the adenine (Ade) moiety relative to the ribose of the Ado ligand. This was resolved with a two-state model in which in the major (0.57 occupancy) conformer the A16(O)-A11-A9(N)-A8 dihedral angle is 1.9 degrees and the Ade is virtually perpendicular to the corrin ring; in the minor conformer, the Ade is tilted down, and this dihedral is -48.7 degrees. The Co-C and axial Co-N bond lengths and the Co-C-C bond angle are quite similar to those in AdoCbl. The corrin ring is considerably flatter than that of AdoCbl, with a fold angle of 11.7 degrees. The molecule was successfully modeled by molecular mechanics (MM), and rotation of the Ado ligand relative to the corrin gave rise to four locally minimum structures with the Ado in the southern, eastern, northern, or western quadrant, with the southern conformation as the global minimum, as is the case with AdoCbl itself. Nuclear Overhauser effects (nOe's) observed by two-dimensional (2D) NMR were incorporated as restraints in molecular dynamics (MD) and simulated annealing (SA) calculations. A MD simulation at 300 K showed that only the southern conformation is populated with the Ado ligand confined to an arc from over C15 to over C12, while the Ade ring oscillates from perpendicular to parallel to the corrin ring. Twenty-seven structures were collected by MD-SA. Most of these annealed into the southern conformation, but examples of the other conformations were also found. The new analogue is a partially active coenzyme for the ribonucleotide reductase from Lactobacillus leichmanii with maximal activity that is 9.7% of that of AdoCbl itself, and a very high Km value (245 microM compared to 0.54 microM for AdoCbl). In addition, the rate constant for enzyme-induced carbon-cobalt bond cleavage of (alpha-ribo)AdoCbl is 160-fold smaller than that for AdoCbl, and only 1/3 as much cob(II)alamin is produced at the active site.

Adenosine↗

Determination of polyamines in human prostate by high-performance liquid chromatography with fluorescence detection.

A high-performance liquid chromatographic method for the determination of polyamines in human prostate has been developed. This method is based on pre-column derivatization with dansyl chloride (Dns-Cl). The derivatives were separated on a muBondapak C18 column (250x4.6 mm I.D.; 10 microm), and eluted with methanol and distilled water using a one-step linear gradient. The column eluate was monitored by fluorescence detection (excitation, 370 nm; emission, 506 nm). The within-assay precision of the study (C.V.) was as follows: putrescine (PUT) 2.88%, spermidine (SPD) 2.94% and spermine (SP) 1.17%. The between-assay precision (C.V.) was: PUT 2.66%, SPD 3.06%, SP 2.79%. The recovery was greater than 97%. The detection limit for PUT, SPD and SP were 0.05, 0.08 and 0.06 nmol/ml, respectively. In contrast to other studies, sample or polyamine derivatives did not require extraction with an organic solvent such as ethanol, evaporation under vacuum or other condensation procedures. This is a simple, rapid and sensitive method that can be applied to the determination of polyamines in nearly all biological tissues and body fluids, such as urine and serum.

Chromatography, High Pressure Liquid↗

Biological behavior of plasmid in Rhizobium sp. strain S25 from Tephrosia candida.

Rhizobium sp. strain S25 was isolated from the nodule on Tephrosia candida in Hainan Province, China. The strain showed high stress tolerance. The plasmid profile of strain S25, examined by the Eckhardt procedure, indicated that the strain harbors only one plasmid with an estimated size of 150 kb. The plasmid was shown to carry nod and nif genes by hybridization with probes of nodABC and nifHDK genes. Plasmid curing was carried out using the Bacillus subtilis sacB to generate derivatives of strain S25. In comparison with the parent strain S25, the cured derivative lost its ability to nodulate the host plant. Loss of the plasmid reduced significantly the strain's tolerance to acid, nitrous, and multiple antibiotics. The properties of the cured strain also indicated that the plasmid was involved in carbon and nitrogen metabolism. Reintroduction of the plasmid from S25 in the cured derivative restored its original biological phenotypes.

DNA, Bacterial↗

Lipid ozonation products activate phospholipases A2, C, and D.

Ozone exposure, in vitro, has been shown to activate phospholipases A2 (PLA2), C (PLC), and D (PLD) in airway epithelial cells. However, because of its high reactivity, ozone cannot penetrate far into the air/lung tissue interface. It has been proposed that ozone reacts with unsaturated fatty acids (UFA) in the epithelial lining fluid (ELF) and cell membranes to generate a cascade of lipid ozonation products (LOP) that mediate ozone-induced toxicity. To test this hypothesis, we exposed cultured human bronchial epithelial cells (BEAS-2B) to LOP (1-100 microM) produced from the ozonation of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) and measured the activity of PLA2, PLC, and PLD. The PLA2 isoform responsible for arachidonic acid release (AA) in stimulated cultures was also characterized. Activation of PLA2, PLC, and PLD by three oxidants, hydrogen peroxide (H2O2), tert-butyl hydroperoxide (t-BOOH) and 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) also was measured and compared to that of LOP. The derivatives of ozonized POPC at the sn-2 residue, 9-oxononanoyl (PC-ALD), 9-hydroxy-9-hydroperoxynonanoyl (PC-HHP), and 8-(-5-octyl-1,2,4-trioxolan-3-yl-) octanoyl (POPC-OZ) selectively activated PLA2 in a dose-dependent fashion. Cytosolic PLA2 (cPLA2) measured in the cytosolic fraction of stimulated cell lysates was found to be the predominant isoform responsible for AA release. PLC activation was exclusively induced by the hydroxyhydroperoxide derivatives. PC-HHP and the 9-carbon hydroxyhydroperoxide (HHP-C9) increased PLC activity. PLD activity also was induced by LOP generated from POPC. Incubation of cultures with H2O2 alone did not stimulate PLC; however, in the presence of the aldehyde, nonanal, a 62 +/- 2% increase in PLC activity was found, suggesting that the increase in activity was due to the formation of the intermediate HHP-C9. t-BOOH, and AAPH also failed to induce PLA2 activation, but did activate PLC, under conditions of exposure identical to that of LOP. Only t-BOOH activated PLD. These results suggest that biologically relevant concentrations of LOP activate PLA2, PLC, and PLD in the airway epithelial cell, a primary target to ozone exposure. The activation of these phospholipases may play a role in the development of lung inflammation during ozone exposure.

Arachidonic Acid↗