Biomedical subjects

L Zhu

Publications and source records attributed to L Zhu.

At least 253 records · Page 14Linked to original sources

PubMed1998

Moho offset across the northern margin of the tibetan plateau

Anomalous double-pulse teleseismic P-wave arrivals were observed at one station near the northern margin of the Tibetan Plateau. The azimuthal dependence of the waveform distortion and its absence at nearby stations indicated that the distortion was produced by receiver-side crustal heterogeneity. Modeling of the three-component data revealed a 15- to 20-kilometer Moho offset that occurs over a narrow lateral range of less than 5 kilometers. This east-west-striking offset separates the thick Tibetan Plateau crust from the Qaidam Basin crust. Such a sharp crustal thickness change implies a weak Tibetan Plateau crust that thickens vertically in response to penetration by India from the south and to blockage caused by a strong Qaidam Basin crust to the north.

Journal Article↗

PubMed1998

Telomere length regulation in mice is linked to a novel chromosome locus.

Little is known about the mechanisms that regulate species-specific telomere length, particularly in mammalian species. The genetic regulation of telomere length was therefore investigated by using two inter-fertile species of mice, which differ in their telomere length. Mus musculus (telomere length >25 kb) and Mus spretus (telomere length 5-15 kb) were used to generate F1 crosses and reciprocal backcrosses, which were then analyzed for regulation of telomere length. This analysis indicated that a dominant and trans-acting mechanism exists capable of extensive elongation of telomeres in somatic cells after fusion of parental germline cells with discrepant telomere lengths. A genome wide screen of interspecific crosses, using M. spretus as the recurrent parent, identified a 5-centimorgan region on distal chromosome 2 that predominantly controls the observed species-specific telomere length regulation. This locus is distinct from candidate genes encoding known telomere-binding proteins or telomerase components. These results demonstrate that an unidentified gene(s) mapped to distal chromosome 2 regulates telomere length in the mouse.

Animals↗

PubMed1998

Construction of a modular yeast two-hybrid cDNA library from human EST clones for the human genome protein linkage map.

Identification of all human protein-protein interactions will lead to a global human protein linkage map that will provide important information for functional genomics studies. The yeast two-hybrid system is a powerful molecular genetic approach for studying protein-protein interactions. To apply this technology to generate a human protein linkage map, the first step is to construct two-hybrid cDNA libraries that cover the entire human genome. With a homologous recombination-mediated approach, we have constructed a modular human EST-derived yeast two-hybrid library in the Gal4 activation domain-based vector, pACT2. Quality analysis of this library indicated that the approach of constructing two-hybrid cDNA libraries from individually arrayed human EST clones is feasible, and such a two-hybrid library is suitable for detecting protein-protein interactions. This is also the first time that a comprehensive two-hybrid system cDNA library has been constructed from a collection of individually arrayed EST clones.

Chromosome Mapping↗

PubMed1998

Molecular defects in ferrochelatase in patients with protoporphyria requiring liver transplantation.

Protoporphyria is a genetic disorder in which a deficiency of mitochondrial ferrochelatase activity causes accumulation of protoporphyrin that produces severe liver damage in some patients. In this study, mutations of the ferrochelatase gene were examined in eight unrelated patients who had liver transplantation. RNA was prepared from liver and/ or lymphoblasts, and specific reverse transcriptase-nested polymerase chain reactions amplified and sequenced ferrochelatase cDNAs. Products shorter than normal resulted from an exon 3 deletion in three patients, exon 10 deletion in two, exon 2 deletion in one, and deletion of five nucleotides in exon 5 in one. Sequence of normal-size products revealed no other mutations. Western blot showed a reduced quantity of normal-size ferrochelatase protein in protoporphyria liver compared with normal liver (19-51%, mean 32% of normal). Levels of the mitochondrial protein F1-ATPase beta-subunit were not decreased to a similar degree. Liver ferrochelatase activity was reduced more than could be explained by the decrease in ferrochelatase protein (4-20%, mean 9% of normal). These results establish genetic heterogeneity in the most severe phenotype of protoporphyria. However, the gene mutations found share the property of causing a major structural alteration in the ferrochelatase protein.

Adolescent↗

PubMed1998

Junctioning of lateral and anterior fields in head and neck cancer: a dosimetric assessment of the monoisocentric technique (including reproducibility).

PURPOSE: The matching or junctioning of two lateral fields with an anterior field is commonly performed in the treatment of head and neck cancer. A monoisocentric technique utilising asymmetric collimation is potentially associated with improved dosimetry in the plane of the junction due to decreased reliance on operator skill and the avoidance of couch movement. The aim of this study was not only to assess the average dose delivered in the plane of the junction, but also the reproducibility of this dose for the monoisocentric technique and two other commonly used techniques. METHODS AND MATERIALS: An anthropomorphic head and neck wax phantom was fashioned to allow the placement of 22 TLD chips in a 2-mm thick transverse plane positioned superior to the potential site of the larynx. Three different treatment techniques were used with the phantom being treated by four different operators a minimum of 20 times for each technique: (1) "straight fields"--using isocentric laterals with an anterior field junctioned in the midline. This technique makes no allowance for divergence; (2) "angled fields"--couch and gantry rotation are used to account for divergence; (3) "monoisocentric"--using asymmetric collimators to create a single isocenter. RESULTS: For an applied dose of 1 Gy the monoisocentric technique produced a mean dose measured of 1.01 Gy compared with 1.23 and 0.92 Gy for techniques 1 and 2. The reproducibility of the mean dose measured was better for the monoisocentric technique by a factor of 2. The superior reproducibility of the monoisocentric technique was not found to be operator dependent. CONCLUSIONS: A monoisocentric technique for the treatment of two laterals and an anterior field in head and neck cancer is likely to be associated with more accurate and reproducible dosimetry in the plane of the junction. Our center has subsequently adopted this technique for matching such fields.

Head and Neck Neoplasms↗

PubMed1998

Non-small cell lung cancer cyclooxygenase-2-dependent regulation of cytokine balance in lymphocytes and macrophages: up-regulation of interleukin 10 and down-regulation of interleukin 12 production.

Tumor-derived prostaglandin E2 (PGE2) modifies cytokine balance and inhibits host immunity. We hypothesized that a high level of PGE2 production by lung tumor cells is dependent on tumor cyclooxygenase (COX)-2 expression. We found that PGE2 production by A549 non-small cell lung cancer (NSCLC) cells was elevated up to 50-fold in response to interleukin (IL)-1beta. Reversal of IL-1beta-induced PGE2 production in A549 cells was achieved by specific pharmacological or antisense oligonucleotide inhibition of COX-2 activity or expression. In contrast, specific COX-1 inhibition was not effective. Consistent with these findings, IL-1beta induced COX-2 mRNA expression and protein production in A549 cells. Specific inhibition of COX-2 abrogated the capacity of IL-1beta-stimulated A549 cells to induce IL-10 in lymphocytes and macrophages. Furthermore, specific inhibition of A549 COX-2 reversed the tumor-derived PGE2-dependent inhibition of macrophage IL-12 production when whole blood was cultured in tumor supernatants. Our results indicate that lung tumor-derived PGE2 plays a pivotal role in promoting lymphocyte and macrophage IL-10 induction while simultaneously inhibiting macrophage IL-12 production. Immunohistochemistry of human NSCLC tissues obtained from lung cancer resection specimens revealed cytoplasmic staining for COX-2 within tumor cells. This is the first description of functional COX-2 expression by NSCLC cells and the definition of a pathway whereby tumor COX-2 expression and a high level of PGE2 production mediate profound alteration in cytokine balance in the lung cancer microenvironment.

Adenocarcinoma↗

PubMed1998

Developmental expression of mouse erythrocyte protein 4.2 mRNA: evidence for specific expression in erythroid cells.

Erythrocyte protein 4.2 (P4.2) is an important component of the erythrocyte membrane skeletal network with an undefined biologic function. Presently, very little is known about the expression of the P4.2 gene during mouse embryonic development and in adult animals. By using the Northern blot and in situ hybridization techniques, we have examined the spatial and temporal expression of the P4.2 gene during mouse development. We show that expression of the mouse P4.2 gene is temporally regulated during embryogenesis and that the P4.2 mRNA expression pattern coincides with the timing of erythropoietic activity in hematopoietic organs. P4.2 transcripts are first detected in embryos on day 7.5 of gestation and are localized exclusively in primitive erythroid cells of yolk sac origin. These erythroid cells remain to be the only source for P4.2 expression until the switch of the hematopoietic producing site to fetal liver. In mid- and late-gestation periods, P4.2 mRNA expression is restricted to the erythroid cells in fetal liver and to circulating erythrocytes. Around and after birth, the site for P4.2 expression is switched from liver to spleen and bone marrow, and P4.2 transcripts are only detected in cells of the erythroid lineage. These results provide the evidence for specific P4.2 expression in erythroid cells. In addition, the timing and pattern of expression of the P4.2 gene suggest the specific regulation of the P4.2 gene.

Animals↗

PubMed1998

DNA vaccines against tuberculosis.

DNA plasmids encoding Mycobacterium tuberculosis antigen 85 (Ag85) were tested as vaccines in animal models. Ag85 DNA induced relevant immune responses (i.e. T helper (Th) cells, Th1 cytokines and cytotoxic T lymphocytes) and was protective in mouse and guinea pig models of mycobacterial disease. Therefore, DNA vaccination holds promise as an effective means of preventing tuberculosis in humans. Furthermore, this technique is amenable to identifying the protective antigens of M. tuberculosis.

Acyltransferases↗

PubMed1998

Blood pressure reduction, cardiovascular diseases, and cognitive decline in the mini-mental state examination in a community population of normal very old people: a three-year follow-up.

We observed the decline in cognitive functioning in a community cohort of 924 persons aged > or =75 years with initially good cognition. Cognitive performance was indexed with the Mini-Mental State Examination (MMSE) on two occasions, 3 years apart. The average decline in MMSE score was 0.4 points per year. However, 23.4% of the participants lost more than 10% of their baseline MMSE scores. Women declined more rapidly than men. Baseline advanced age, lower education, and stroke predicted the greater decline in women. It appears that the relation of lower education and stroke with cognitive decline was more pronounced in men, but age did not predict cognitive decline in men. In addition, there was a correlation between systolic pressure reduction and cognitive decline in women, which was not accounted for by other factors. In summary, although the cognitive ability is generally spared longitudinally, in terms of the entire population, a considerable proportion of individuals show substantial decline that is related to several factors such as advanced age, female gender, lower education, stroke, and systolic pressure reduction.

Aged↗

PubMed1998

Comparison of 15N- and 13C-determined parameters of mobility in melittin.

Backbone and tryptophan side-chain mobilities in the 26-residue, cytolytic peptide melittin (MLT) were investigated by 15N and 13C NMR. Specifically, inverse-detected 15N T1 and steady-state NOE measurements were made at 30 and 51 MHz on MLT at 22 degrees C enriched with 15N at six amide positions and in the Trp19 side chain. Both the disordered MLT monomer (1.2 mM peptide at pH 3.6 in neat water) and alpha-helical MLT tetramer (4.0 mM peptide at pH 5.2 in 150 mM phosphate buffer) were examined. The relaxation data were analyzed in terms of the Lipari and Szabo model-free formalism with three parameters: tau m, the correlation time for the overall rotation; S2, a site-specific order parameter which is a measure of the amplitude of the internal motion; and tau e, a local, effective correlation time of the internal motion. A comparison was made of motional parameters from the 15N measurements and from 13C measurements on MLT, the latter having been made here and previously [Kemple et al. (1997) Biochemistry, 36, 1678-1688]. tau m and tau e values were consistent from data on the two nuclei. In the MLT monomer, S2 values for the backbone N-H and C alpha-H vectors in the same residue were similar in value but in the tetramer the N-H order parameters were about 0.2 units larger than the C alpha-H order parameters. The Trp side-chain N-H and C-H order parameters, and tau e values were generally similar in both the monomer and tetramer. Implications of these results regarding the dynamics of MLT are examined.

Amino Acid Sequence↗

PubMed1998

A NOESY-HSQC simulation program, SPIRIT.

A program SPIRIT (Simulation Program considering Incomplete Recovery of z magnetization and INEPT Transfer efficiency) has been developed to simulate three-dimensional NOESY-HSQC spectra. This program takes into account (1) different transfer efficiency during INEPT and reverse INEPT durations due to differential relaxation rates and 1J coupling constants; (2) the different effect of the sensitivity-enhancement scheme on CH, CH2 and CH3 systems; and (3) incomplete recovery of longitudinal magnetization between scans. The simulation program incorporates anisotropic tumbling mode for symmetric tops, and allows for differential external relaxation rates for protons. Some well-defined internal motions, such as the fast rotation of methyl groups, are taken into account. The simulation program also allows for input of multiple conformations and their relative populations to calculate the average relaxation matrix to account for slow internal motions. With the SPIRIT program, the sensitivity-enhanced NOESY-HSQC experiment can be used directly in the evaluation of the accuracy of structures, which can potentially be improved by direct refinement against the primary data.

Computer Simulation↗

PubMed1998

Using the yeast two-hybrid system to identify human epithelial cell proteins that bind gonococcal Opa proteins: intracellular gonococci bind pyruvate kinase via their Opa proteins and require host pyruvate for growth.

Neisseria gonorrhoeae opacity-associated (Opa) proteins are a family of outer membrane proteins involved in gonococcal adherence to and invasion of human cells. We wanted to identify additional roles for Opa in the infectious process and used the yeast two-hybrid system to identify human epithelial cell proteins that interact with Opa proteins. Although this system has been used successfully to identify many types of interacting proteins, it has not been used to screen a human cell cDNA library for binding partners of a prokaryotic outer membrane protein. Therefore, we were also interested in exploring the versatility of the yeast two-hybrid system in identifying bacteria-host interactions. Using OpaP from strain F62SF as bait, we screened a HeLa cell cDNA library for Opa-interacting proteins (OIPs). We identified five different OIPs, designated OIP1-OIP5, two of which are homologous to human proteins--thyroid hormone receptor interacting protein (TRIP6) and pyruvate kinase isoenzyme M2 (PK). In the studies presented here, we investigated the interaction between Opa proteins and PK in more depth. Opa-PK interactions were confirmed by in vitro and in vivo assays independent of the yeast two-hybrid system. Escherichia coli expressing six different Opa proteins from gonococcal strain FA1090 all bound more PK than Opa-negative E. coli in in vitro binding assays. Using anti-PK antibody and fluorescence microscopy, we showed that human epithelial cell PK co-localizes with intracellular Opa+ gonococci and E. coli expressing Opa proteins. Using a mutant of N. gonorrhoeae unable to grow on pyruvate or lactate, it appears that intracellular pyruvate is essential for gonococcal growth and survival. These results suggest a novel mechanism in bacterial pathogenesis, i.e. the requirement for direct molecular interaction with a host metabolic enzyme (PK) for the acquisition of an essential intracellular carbon source and growth substrate (pyruvate). These results demonstrate that the yeast two-hybrid system is a valuable tool for identifying biologically relevant interactions between bacteria and host proteins, providing valuable leads for further investigations into novel mechanisms of bacterial pathogenesis.

Amino Acid Sequence↗

PubMed1998

Complexity in uterine macrophage responses to cytokines in mice.

Uterine stromal macrophages change dramatically in density and morphology through the estrous cycle and during early pregnancy, whereas those in the mesometrial triangle do not undergo these changes. The mononuclear phagocytic growth factor, colony-stimulating factor-1 (CSF-1), regulates both the density and morphology of uterine macrophage populations, as shown by the fact that uterine macrophages are depleted and more rounded in the absence of CSF-1 caused by the osteopetrotic (csfm(op)) null mutation, compared to those of normal mice. Restoration of circulating CSF-1 to the nullizygous mice did not affect stromal macrophage density although it restored the population in the mesometrial triangle. This suggests CSF-1 regulation of these macrophage populations by local and humoral routes, respectively. Nevertheless, even in the absence of CSF-1, stromal macrophage population density varies 30-fold through the estrous cycle, suggesting the involvement in their regulation of factors other than CSF-1, such as the chemokines, which are chemoattractive for macrophages. The mRNA for the chemokines JE (MCP-1), C10, RANTES, and MIP1alpha are expressed in the uterus, with elevated levels observed on the first day of pregnancy. Such molecules, together with CSF-1, may play a role in modulating the complexities of uterine macrophage dynamics in response to sex steroid hormones and mating.

Animals↗

PubMed1998

Modeling human eye aberrations and their compensation for high-resolution retinal imaging.

We introduced a mathematical eye model using Gullstrand's six-surface eye model modified by clinically measured aspherical data to study human eye aberrations and their compensation for high-resolution retinal imaging. Ray tracing was used to characterize aberrations and point spread functions (PSFs) of the eye model. By using the Zernike polynomial decomposition of the calculated pupil function, we quantified the wavefront aberrations. Based on calculated PSFs, we designed optical inverse filters to reduce the aberrations for a large pupil size and improve the resolution. Spherical aberration and oblique astigmatism were found to be in good agreement with published experimental measurements. Spherical aberration and defocus were the most significant aberrations for on-axis imaging, whereas oblique astigmatism and coma combined with spherical aberration and defocus were most significant for off-axis imaging. The best retinal image resolution occurred at 2- to 3-mm pupil diameter. After aberration correction for an 8-mm diameter pupil, the resolutions for on-axis or 9 degrees off-axis imaging points were very close to diffraction-limited resolutions. Over a limited field of view (FOV), retinal image resolution of the eye model can be greatly improved by aberration correction using aspheric and astigmatic lenses. For imaging large FOVs, space-variant compensation techniques will be required for aberration correction.

Astigmatism↗

PubMed1998

Quantification of the 3-D electromagnetic power absorption rate in tissue during transurethral prostatic microwave thermotherapy using heat transfer model.

Experiments were performed in a tissue microwave-equivalent phantom gel to quantitatively examine the volumetric heating produced by a microwave antenna with a peripheral cooling system for the transurethral prostatic thermotherapy. Based on previous research, expression for the specific absorption rate (SAR) of microwave energy in the gel was extended to three dimensions, which includes its dependence on radial, angular, and axial direction. A theoretical heat transfer model was developed to study the temperature distribution in the gel by introducing this proposed SAR expression. The parameters in this expression and the convection coefficient due to the chilled water running around the antenna were determined using a least-square residual fit of the theoretical temperature predictions to the experimentally measured steady-state temperature field within the gel. The analytical expression of the three-dimensional SAR distribution obtained in this study will help provide a better understanding of the microwave heating pattern in the prostatic tissue and, thus, to aid in designing improved applicators. It can also be used in the future as an accurate input to heat transfer models which predict temperature distributions during the transurethral microwave thermotherapy.

Gels↗

PubMed1998

Oligoribonuclease is encoded by a highly conserved gene in the 3'-5' exonuclease superfamily.

Oligoribonuclease, a 3'-to-5' exoribonuclease specific for small oligoribonucleotides, was purified to homogeneity from extracts of Escherichia coli. The purified protein is an alpha2 dimer of 40 kDa. NH2-terminal sequence analysis of the protein identified the gene encoding oligoribonuclease as yjeR (o204a), a previously reported open reading frame located at 94 min on the E. coli chromosome. However, as a consequence of the sequence information, the translation start site of this open reading frame has been revised. Cloning of yjeR led to overexpression of oligoribonuclease activity, and interruption of the cloned gene with a kanamycin resistance cassette eliminated the overexpression. On the basis of these data, we propose that yjeR be renamed orn. Orthologs of oligoribonuclease are present in a wide range of organisms, extending up to humans.

Bacterial Proteins↗

PubMed1998

Functional domains in the retroviral transmembrane protein.

The envelope glycoproteins of the mammalian type C retroviruses consist of two subunits, a surface (SU) protein and a transmembrane (TM) protein. SU binds to the viral receptor and is thought to trigger conformational changes in the associated TM protein that ultimately lead to the fusion of viral and host cell membranes. For Moloney murine leukemia virus (MoMuLV), the envelope protein probably exists as a trimer. We have previously demonstrated that the coexpression of envelope proteins that are individually defective in either the SU or TM subunits can lead to functional complementation (Y. Zhao et al., J. Virol. 71:6967-6972, 1997). We have now extended these studies to investigate the abilities of a panel of fusion-defective TM mutants to complement each other. This analysis identified distinct complementation groups within TM, with implications for interactions between different regions of TM in the fusion process. In viral particles, the C-terminal 16 amino acids of the MoMuLV TM (the R peptide) are cleaved by the viral protease, resulting in an increased fusogenicity of the envelope protein. We have examined the consequences of R peptide cleavage for the different TM fusion mutants and have found that this enhancement of fusogenicity can only occur in cis to certain of the TM mutants. These results suggest that R peptide cleavage enhances the fusogenicity of the envelope protein by influencing the interaction of two distinct regions in the TM ectodomain.

3T3 Cells↗

PubMed1998

Requirement of cyclin E-Cdk2 inhibition in p16(INK4a)-mediated growth suppression.

Loss-of-function mutations of p16(INK4a) have been identified in a large number of human tumors. An established biochemical function of p16 is its ability to specifically inhibit cyclin D-dependent kinases in vitro, and this inhibition is believed to be the cause of the p16-mediated G1 cell cycle arrest after reintroduction of p16 into p16-deficient tumor cells. However, a mutant of Cdk4, Cdk4(N158), designed to specifically inhibit cyclin D-dependent kinases through dominant negative interference, was unable to arrest the cell cycle of the same cells (S. van den Heuvel and E. Harlow, Science 262:2050-2054, 1993). In this study, we determined functional differences between p16 and Cdk4(N158). We show that p16 and Cdk4(N158) inhibit the kinase activity of cellular cyclin D1 complexes through different mechanisms. p16 dissociated cyclin D1-Cdk4 complexes with the release of bound p27(KIP1), while Cdk4(N158) formed complexes with cyclin D1 and p27. In cells induced to overexpress p16, a higher portion of cellular p27 formed complexes with cyclin E-Cdk2, and Cdk2-associated kinase activities were correspondingly inhibited. Cells engineered to express moderately elevated levels of cyclin E became resistant to p16-mediated growth suppression. These results demonstrate that inhibition of cyclin D-dependent kinase activity may not be sufficient to cause G1 arrest in actively proliferating tumor cells. Inhibition of cyclin E-dependent kinases is required in p16-mediated growth suppression.

Animals↗