Biomedical subjects
Publications and source records attributed to B Lai.
Intracellular granules containing ferric and ferrous iron formed in Shewanella putrefaciens CN32 during dissimilatory reduction of solid-phase ferric iron. It is the first in situ detection at high resolution (150 nm) of a mixed-valence metal particle residing within a prokaryotic cell. The relationship of the internal particles to Fe(III) reduction may indicate a respiratory role.
We have probed the effects of transverse variations in pinning strength on charge-density-wave (CDW) structure in NbSe3 by x-ray micro-beam diffraction. In ribbonlike crystals having a large longitudinal step in thickness, the CDW first depins on the thick side of the step, causing rotations of the CDW wave vector. By measuring these rotations as a function of position and electric field, the corresponding shear strains are determined, allowing the CDW's shear modulus to be estimated. These results demonstrate the usefulness of x-ray microdiffraction as a tool in studying collective dynamics in electronic crystals.
An X-ray micro/nanodiffraction technique that allows structural characterization of individual nanomaterials has been developed at an insertion-device beamline of the Advanced Photon Source. Using the extremely high brightness of the third-generation synchrotron radiation source and advanced high-resolution high-energy zone-plate focusing optics, X-rays of energies from 6 to 12 keV have been focused into a spot smaller than 200 nm with a photon density gain of more than 50,000 so that significant photon flux can be intercepted by a nanoscale material to generate a measurable diffraction signal for structural characterization. This paper describes the instrumentation of the technique and discusses the application of the technique to studies of tin oxide nanobelts.
Phase-contrast X-ray diffraction imaging and high-angular-resolution diffraction combined with phase-contrast radiographic imaging were employed to characterize defects and perfection of a uniformly grown tetragonal lysozyme crystal in the symmetric Laue case. The full-width at half-maximum (FWHM) of a 4 4 0 rocking curve measured from the original crystal was approximately 16.7 arcsec and imperfections including line defects, inclusions and other microdefects were observed in the diffraction images of the crystal. The observed line defects carry distinct dislocation features running approximately along the <1 1 0> growth front and have been found to originate mostly in a central growth area and occasionally in outer growth regions. Inclusions of impurities or formations of foreign particles in the central growth region are resolved in the images with high sensitivity to defects. Slow dehydration led to the broadening of a fairly symmetric 4 4 0 rocking curve by a factor of approximately 2.6, which was primarily attributed to the dehydration-induced microscopic effects that are clearly shown in X-ray diffraction images. The details of the observed defects and the significant change in the revealed microstructures with drying provide insight into the nature of imperfections, nucleation and growth, and the properties of protein crystals.
Explore the source record for details and available documents.
A measurement of the horizontal coherence function of 7.9 keV radiation from an undulator beam line at the Advanced Photon Source is reported. X-ray diffraction from a phase-shifting mask was used, and the coherence function was measured as a function of the width of beam-conditioning slits in the beam line. The coherence distribution is found to be best described by a Lorentzian function.
Columnar microstructure in step-graded Si(1-x)Ge(x)/Si(001) structures with low threading dislocation densities has been determined using high angular resolution (approximately 0.005 degrees ) x-ray microdiffraction. X-ray rocking curves of a 3-microm-thick strain-relaxed Si(0.83)Ge(0.17) film show many sharp peaks and can be simulated with a model having a set of Gaussians having narrow angular widths (0.013 degrees -0.02 degrees ) and local ranges of tilt angles varying from 0.05 degrees to 0.2 degrees. These peaks correspond to individual tilted rectangular columnar micrograins having similar (001) lattice spacings and average areas of 0.8 to 2.0 microm(2).
Magnetic x-ray diffraction combined with x-ray focusing optics was used to image individual antiferromagnetic spin density wave domains in a chromium single crystal at the micron scale. The cross section for nonresonant magnetic x-ray scattering depends on the antiferromagnetic modulation vector and spin polarization direction and allows these quantities to be extracted independently. The technique was used to show that the broadening of the nominally first-order "spin-flip" transition at 123 kelvin, at which the spins rotate by 90 degrees C, originates at the walls between domains with orthogonal modulation vectors. During cooling, the transition begins at these walls and progresses inward. The modulation vector domains are themselves unchanged.
Explore the source record for details and available documents.
Conventional x-ray diffraction topography is currently used to map defects in the bulk of protein crystals, but the lack of sufficient contrast is frequently a limiting factor. We experimentally demonstrate that this barrier can be circumvented using a method that combines phase sensitive and diffraction imaging principles. Details of defects revealed in tetragonal lysozyme and cubic ferritin crystals are presented and discussed. The approach enabling the detection of the phase changes of diffracted x rays should prove to be useful in the study of defect structures in a broad range of biological macromolecular crystals.
OBJECTIVE: Few previous studies have shown high frequency of loss of heterozygosity (LOH) on chromosome 4q in hepatocellular carcinoma (HCC). We define more clearly the deletion regions that may harbor the putative tumor suppressor genes in HCC. METHODS: Forty-eight cases of HCC and their corresponding non-tumor liver tissues were investigated with 12 microsatellite polymorphic markers for LOH. RESULTS: Twenty-one of 48 (44%) and 30 of 48 (63%) tumors showed LOH on at least one locus on the short and the long arms respectively. Two distinct common deleted regions (CDR) with different patterns of deletion were identified. The first CDR was located on 4q22-q25, between loci D4S392 and D4S1625. In addition, 17 of 27 (63%) of the informative tumors showed LOH on this region with locus D4S406 and constituted the highest rate of LOH on chromosome 4. The second CDR was located at 4q27-q31, between loci D4S1625 and D4S1652. CONCLUSION: There are at least two tumor suppressor loci on 4q.
OBJECTIVE: To investigate the anti-tumor effects of human single chain interleukin-12 (hscIL-12). METHOD: pcDNA/hscIL-12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin method. The 7721/hscIL-12 cells which secrete hscIL-12 stably, were obtained via G418 selection, and in vitro the influence of hscIL-12 gene transduction on the growth of tumor cells was evaluated by cell cycle analysis. In vivo, genetically engineered 7721 cells (7721/hscIL-12, 7721/pcDNA) and parental cells were implanted into BALB/c nude mice, respectively. 7721/pcDNA and 7721/hscIL-12 groups were divided into two sub-groups on day 8: one was administered with hPBL twice, 6 days at interval; the other was given equal volume of PBS. Mice were sacrificed on day 26, and spleens and tumors were taken out for histologic assay. RESULTS: hscIL-12 produced stably by 7721/hscIL-12 cells had bioactivity, and it was proved by Western blot, immunocytochemistry, and in situ hybridization. In vitro, compared with 7721 and 7721/pcDNA, the 7721/hscIL-12 grew much more slowly. FACS assay showed apparent G1 arrest of 7721/hscIL-12 cells. In animal experiment, on day 8 after inoculation, the tumors of 7721 and 7721/pcDNA group were up to 5 approximately 7 mm, while those of 7721/hscIL-12 group were 2 approximately 4 mm. When treated with hPBL, the tumor of 7721/hscIL-12 group disappeared completely. Histologically, the tumors from 7721/hscIL-12 without hPBL treatment had numerous lymphocyte infiltration, the tumor cells displayed depression looking, atrophy, focal necrosis and apoptosis, whereas the tumors of 7721 and 7721/pcDNA groups grew thrivingly. CONCLUSION: hscIL-12 transduced 7721 cells could induced significant antitumor immune response which resulted in tumor regression totally when the hPBL was inoculated, and also hscIL-12 has certain effects on mice immune system. These findings suggest that hscIL-12 and hscIL-12 gene therapy might have promising prospects in clinical application.
Abundant, micrometer-scale, spherical aggregates of 2- to 5-nanometer-diameter sphalerite (ZnS) particles formed within natural biofilms dominated by relatively aerotolerant sulfate-reducing bacteria of the family Desulfobacteriaceae. The biofilm zinc concentration is about 10(6) times that of associated groundwater (0.09 to 1.1 parts per million zinc). Sphalerite also concentrates arsenic (0.01 weight %) and selenium (0.004 weight %). The almost monomineralic product results from buffering of sulfide concentrations at low values by sphalerite precipitation. These results show how microbes control metal concentrations in groundwater- and wetland-based remediation systems and suggest biological routes for formation of some low-temperature ZnS deposits.
Studies on the influence of organic cosolvents on lysozyme folding have been reported. As most of the researches are confined to a few specific molecules and focus on equilibrium states, less is known about the effect on folding dynamics. We have studied the influence of six soluble organic cosolvents on hen egg white lysozyme heat induced denaturation and refolding dynamics. It was found that trifluoroethanol (TFE) can change the folding pathway significantly. With the presence of TFE, the overshot phenomenon generally observed in lysozyme folding at 222 nm disappears. The common mechanism of how organic cosolvents influence folding is analyzed. The heat induced denaturation temperature was found to have a quantitative relationship with the slow phase rate constant during folding. We discuss this finding and hypothesize that it is due to the similar influence of organic cosolvent on the transition state of heat denaturation and refolding.
Explore the source record for details and available documents.
X-ray fluorescence microscopy and microspectroscopy with micrometre spatial resolution and unprecedented capabilities for the study of biological and environmental samples are reported. These new capabilities are a result of both the combination of high-brilliance synchrotron radiation and high-performance X-ray microfocusing optics and the intrinsic advantages of X-rays for elemental mapping and chemical-state imaging. In this paper, these capabilities are illustrated by experimental results on hard X-ray phase-contrast imaging, X-ray fluorescence (XRF) imaging and microspectroscopy of mycorrhizal plant roots and fungi in their natural hydrated state. The XRF microprobe is demonstrated by the simultaneous mapping of the elemental distributions of P, S, K, Ca, Mn, Fe, Ni, Cu and Zn with a spatial resolution of approximately 1 x 3 micron and with an elemental sensitivity of approximately 500 p.p.b. Microspectroscopy with the same spatial resolution is demonstrated by recording near-edge X-ray absorption (XANES) spectra of Mn at a concentration of approximately 3 p.p.m.
OBJECTIVE: To study the effects of exogenous wild type p53 suppressor gene on malignant growths of human lung cancer cell line with mutant type p53 gene. METHOD: Four human lung cancer cell line were screened for mutations in the exon 5 through exon 8 of the p53 tumor suppressor gene with immunohistochemistry, polymerase chain reaction (PCR)/single strand conformation polymorphism (SSCP) and DNA sequence analysis. The recombinent plasmid PZIPneoSV-p53 was constructed, which express wild type p53 gene. A transfected cell line, 801-D-p53, was obtained after transferred the plasmid into 801-D cell line by gene gun mediated and selected by G418. The exogenous p53 in the transfected cell line 801-D-p53 were inspected with PCR, and the alteration of growths of the transfected cell line in vitro and in vivo was observed. RESULT: The point mutation were CGG to CTT transversion at codon 248 in exon 7 was found in human lung cancer cell line 801-D by PCR-SSCP and DNA sequence analysis and nuclear accumulation of the p53 protein was observed. The neo gene and exogenous wild type p53 gene were detected in the transfected cell line 801-D-p53. The cell growth experiment in vitro showed that the parent cell line 801-D growth was very fast, from 1 x 10(5)/ml to 2.5 x 10(5)/ml within 6 days, the transfected PZIP-neo-SV cell line (plasmid without p53) growth as fast as 801-D, but the transfected cell line 801-D-p53 growth was inhibited seriously. The clonogenic formation rate of the parent cell line 801-D, transfected cell line 801-D-PZIP and the transfected cell line 801-D-p53 were 11.2%, 11.4% and 0.46% respectively. The clononogenic formation inhibition rate of the transfected 801-D-p53 was 96% comparing with the parent 801-D cell line. In the experiment of xenogenic tumor transplantation, each cell line was injected subcutaneously into four mice and the growth of xenogenic tumor transplant in nude mice was observed. xenograft growth in all 4 mice in both 801-D and 801-D-PZIP groups, but only 1 xenograft growth among 4 mice of 801-D-p53 group during 2 months. The average volume of xenogenic tumor transplant of 801-D and 801-D-PZIP groups were 6.500 cm3 and 2.231 cm3 respectively and the only xenograft volume of 801-D-p53 group was 0.940 cm3. The tumorigenicity of 801-D-p53 in nude mice was significantly suppressed. CONCLUSION: Exogenous wild type p53 gene may stably exist in the human lung cancer cell line with mutant type p53 after plasmid transfection and suppressed the malignant growth of the transfected cell line 801-D-p53 in vitro and in vivo. These results indicate that the recombinent plasmid expressing wile type p53 may be useful for gene therapy of human lung cancer.
Among the numerous monoclonal antibodies generated against human sperm antigens, HS-13 monoclonal antibody was shown to react with an intra-acrosomal antigen from human, mouse and rat. In this study, HS-13 was used as the affinity ligand for the purification of the cognate antigen from human sperm by immunoaffinity chromatography. The purified cognate antigen from human sperm, designated as HSAg-13, was found to be a protein with a molecular weight of approximately 80 kDa on SDS-PAGE in the presence of reducing reagents. This monoclonal antibody was used as the probe to study the tissue distributions and developmental expression of the cognate antigen from human, mouse and rat by immunohistochemical assays. It was concluded that the antigen recognized by HS-13 antibody is highly sperm specific and found only in sperm and mature testis, but not in any other somatic tissues examined in human and mouse. The antigen was shown to be expressed at the postmeiotic stages of spermatogenesis in mouse and rat. By using indirect immunofluorescent staining assay, HS-13 was shown to react only with the methanol-fixed acrosome-intact sperm but not with the live sperm. Following calcium ionophore A23187 treatment, acrosome-reacted sperm showed either negative staining or residual staining in the equatorial region, suggesting the intra-acrosomal location of HSAg-13. The spontaneous acrosome reaction following overnight incubation in BWW medium resulted in a statistically significant decrease of antibody-stained human sperm. In view of excellent correlations for the scoring of acrosome-intact sperm with that of fluorescence-labeled Pisum sativum agglutinin (PSA) probe, HS-13 monoclonal antibody can be routinely used for monitoring sperm capacitation and acrosome reaction.