Biomedical subjects

S Xu

Publications and source records attributed to S Xu.

At least 73 records · Page 4Linked to original sources

PubMed2001

Restriction enzyme BsoBI-DNA complex: a tunnel for recognition of degenerate DNA sequences and potential histidine catalysis.

BACKGROUND: Restriction endonucleases form a diverse family of proteins with substantial variation in sequence, structure, and interaction with recognition site DNA. BsoBI is a thermophilic restriction endonuclease that exhibits both base-specific and degenerate recognition within the sequence CPyCGPuG. RESULTS: The structure of BsoBI complexed to cognate DNA has been determined to 1.7 A resolution, revealing several unprecedented features. Each BsoBI monomer is formed by inserting a helical domain into an expanded EcoRI-type catalytic domain. DNA is completely encircled by a BsoBI dimer. Recognition sequence DNA lies within a 20 A long tunnel of protein that excludes bulk solvent. Interactions with the specific bases are made in both grooves through direct and water-mediated hydrogen bonding. Interaction with the degenerate position is mediated by a purine-specific hydrogen bond to N7, ensuring specificity, and water-mediated H bonding to the purine N6/O6 and pyrimidine N4/O4, allowing degeneracy. In addition to the conserved active site residues of the DX(n)(E/D)ZK restriction enzyme motif, His253 is positioned to act as a general base. CONCLUSIONS: A catalytic mechanism employing His253 and two metal ions is proposed. If confirmed, this would be the first example of histidine-mediated catalysis in a restriction endonuclease. The structure also provides two novel examples of the role of water in protein-DNA interaction. Degenerate recognition may be mediated by employing water as a hydrogen bond donor or acceptor. The structure of DNA in the tunnel may also be influenced by the absence of bulk solvent.

Catalysis↗

PubMed2001

Flow-injection chemical vapor-generating procedure for the determination of Au by atomic absorption spectrometry.

Volatile Au species in an acidified medium were generated at room temperature by reduction with NaBH4 in acidified aqueous medium using a flow-injection chemical vapor-generation atomic absorption spectrometric (FI-CVG-AAS) system in the presence of micro amounts of sodium diethyldithiocarbamate (DDTC). Precision of 2.0% RSD (n = 11, 2.0 mg L(-1) level) was obtained at a sample throughput of 180 h(-1). A detection limit of 24 ng mL(-1) (3sigma) was obtained with 300 microL sample solution. The method was used for the determination of gold in ore sample digests, and the results obtained agreed well with those obtained by flame AAS.

Journal Article↗

PubMed2001

Over-expression of beta-1,4-galactosyltransferase I, II, and V in human astrocytoma.

PURPOSE: beta-1,4-Galactosyltransferase (beta-1,4-GalT) I, II, and V are the enzymes responsible for the biosynthesis of N-acetyllactosamine on N-glycans by transferring UDP-galactose to the terminal N-acetylglucosamine (N-GlcNAc) residues with the formation of a beta-1,4-linkage. Neoplasms undergo various changes in the carbohydrate of their glycoconjugates, indicating the possible changes in glycosyltransferases themselves. METHOD: Therefore, we compared the expression of beta-1,4-GalTs between astrocytoma and normal brain tissues. RESULTS: Our reverse-transcription polymerase chain reaction (RT-PCR) results showed that beta-1,4-GalT I transcript was absent in normal adult brain but detectable in grade II, III, and IV astrocytomas; the level of beta-1,4-GalT II transcript was increased in grade II, III, and IV astrocytomas while only a trace amount was found in normal brain; beta-1,4-GalT V transcript existed in normal brain and increased in the process of astrocytoma progress, with the highest level in grade IV astrocytoma. By Ricinus communis agglutinin-1 (RCA-1) lectin blot assay, we also found the more extensive galactosylated bands in astrocytomas compared with normal brain. A major 61kD protein was galactosylated in astrocytoma but not in normal brain tissues. CONCLUSION: These results indicate that the increase of galactosylation in astrocytomas may be caused by the alterations of gene expression of beta-1,4-GalT I, II, and V and that the malignant degree of astrocytoma is correlated with the expression of beta-1,4-GalT V.

Astrocytoma↗

PubMed2001

The assembly of amyloidogenic yeast sup35 as assessed by scanning (atomic) force microscopy: an analogy to linear colloidal aggregation?

Amyloidosis is a class of diseases caused by protein aggregation and deposition in various tissues and organs. In this paper, a yeast amyloid-forming protein Sup35 was used as a model for understanding amyloid fiber formation. The dynamics of amyloid formation by Sup35 were studied with scanning force microscopy. We found that: 1) the assembly of Sup35 fibers begins with individual NM peptides that aggregate to form large beads or nucleation units which, in turn, form dimers, trimers, tetramers and longer linear assemblies appearing as a string of beads; 2) the morphology of the linear assemblies differ; and 3) fiber assembly suggests an analogy to the aggregation of colloidal particles. A dipole assembly model is proposed based on this analogy that will allow further experimental testing.

Amyloid↗

PubMed2001

The mechanism of oxidation-induced low-density lipoprotein aggregation: an analogy to colloidal aggregation and beyond?

Atherosclerosis is a disease initiated by lipoprotein aggregation and deposition in artery walls. In this study, the de novo low-density lipoprotein aggregation process was examined. Nine major intermediates were identified in two stages of the aggregation process. In the aggregation stage, low-density lipoprotein molecules aggregate and form nucleation units. The nucleation units chain together and form linear aggregates. The linear aggregates branch and interact with one another, forming fractals. In the fusion stage, spatially adjacent nucleation units in the fractal fuse into curved membrane surfaces, which, in turn, fuse into multilamellar or unilamellar vesicles. Alternatively, some adjacent nucleation units in the fractals assemble in a straight line and form rods. Subsequently, the rods flatten out into rough and then into smooth ribbons. Occasionally, tubular membrane vesicles are formed from the fractals. The aggregation stage seems to be analogous to colloidal aggregation and amyloid fiber formation. The fusion stage seems to be characteristic of the lipid-rich lipoproteins and is beyond colloidal aggregation and amyloid fiber formation.

Amyloid↗

PubMed2001

Fabrication of nanometer-sized protein patterns using atomic force microscopy and selective immobilization.

A new methodology is introduced to produce nanometer-sized protein patterns. The approach includes two main steps, nanopatterning of self-assembled monolayers using atomic force microscopy (AFM)-based nanolithography and subsequent selective immobilization of proteins on the patterned monolayers. The resulting templates and protein patterns are characterized in situ using AFM. Compared with conventional protein fabrication methods, this approach is able to produce smaller patterns with higher spatial precision. In addition, fabrication and characterization are completed in near physiological conditions. The adsorption configuration and bioreactivity of the proteins within the nanopatterns are also studied in situ.

Adsorption↗

PubMed2001

Effect of five flavonoid compounds isolated from Quercus dentata Thunb on superoxide generation in human neutrophils and phosphorylation of neutrophil proteins.

Effect of five yellow compounds isolated from Quercus dentata on superoxide generation and protein phosphorylation in human neutrophils was investigated. The five yellow compounds examined were Kaempferol 3-O-beta-D-glucopyranoside (B), quercetin 3-O-beta-D-glucopyranoside (DA), Kaempferol 3-O-(6"-trans-p-coumaroyl)-beta-D-glucopyranoside (D1), Kaempferol 3-O-(2"-6"-di-trans-p-coumaroyl)-beta-D-glucopyranoside (D7) and Kaempferol 3-O-(2",4"-di-acetyl-3"-cis-p-coumaroyl-6"-trans-p-coumaroyl)-beta-D-glucopyranoside (A). D7 suppressed significantly the superoxide generation induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP). D1 and DA suppressed significantly the superoxide generation induced by arachidonic acid (AA). However, the superoxide generation induced by phorbol 12-myristate 13-acetate (PMA) was suppressed by all yellow compounds. When the cells were incubated with fMLP and D7, the tyrosyl phosphorylation of 67, 60, 58 and 38 kDa proteins of the cells was markedly decreased in a concentration-dependent manner.

Flavonoids↗

PubMed2001

Pattern visual evoked potential in the diagnosis of functional visual loss.

OBJECTIVE: To study the pattern visual evoked potential (P-VEP) in the diagnosis of functional visual loss. STUDY DESIGN: Retrospective study of observational case series. PARTICIPANTS: Seventy-two subjects whose best corrected visual acuity (VA) was 20/50 or worse, with or without visual field defect, and whose visual abnormalities could not be explained by the findings of ophthalmologic and neurologic examination were included in this study. MAIN OUTCOME MEASURES: To compare the P-VEP estimated acuity to the initial subjective VA and to the best-performed VA. RESULTS: Seventy-two subjects with functional visual loss had normal P-VEPs. The initial subjective VA was 20/50 in 9 subjects and < or =20/200 in 42 subjects. After clinical examination and reassurance, the best-performed VA was > or =20/50 in 53 subjects and < or =20/200 in 8 subjects. The discrepancy between the P-VEP estimated acuity and the best-performed VA was less than 3 lines of Snellen acuity in 63 of 72 (87.5%) subjects and more than 4 lines in 6 subjects. These six subjects were three women with loss of vision of unknown origin and three men with injury-related visual loss. CONCLUSIONS: P-VEP has the advantage of objectively predicting VA and is a useful test in the diagnosis of functional visual loss.

Adolescent↗

PubMed2001

Heterogeneous matrix diffusion in crystalline rock--implications for geosphere retardation of migrating radionuclides.

As a basis for an analysis of the effect of rock heterogeneity on radionuclide migration in a single fracture, the geostatistics of the main properties governing solute transport in crystalline rock have been determined experimentally for two granitic rock types. The rock samples were collected at the Aspö Hard Rock Laboratory, Sweden and used to deduce the auto-covariance functions for the porosity, effective diffusivity and partition coefficient, kd, and adsorption kinetics. One-dimensional analytical solutions for the mean values of the temporal moments of the residence time probability density function (PDF) show that the heterogeneity of the rock properties can have a substantial impact on the transport. A case study of the effect of heterogeneity in matrix diffusion for a single fracture could be performed by decomposing the transport problem into a one-dimensional mass transfer problem and a two-dimensional flow problem using a Lagrangian method of description. Monte Carlo simulations of the flow field indicate that the correlation length of the aperture is much longer along the trajectory paths than along an arbitrary direction. Increasing the correlation lengths and variances of the aperture and matrix diffusion increases significantly the variance of the travel time PDF.

Diffusion↗

PubMed2001

Acute effects of whole-body exposure to static magnetic fields and 50-Hz electromagnetic fields on muscle microcirculation in anesthetized mice.

Acute microhemodynamic effects of static and alternating magnetic fields at a threshold level were investigated on modulating the muscle capillary mirocirculation in pentobarbital-anesthetized mice. The skin in a tibialis anterior was circularly removed with 1.5 mm diameter for intravital-microscopic recording of the capillary blood velocity in the tibialis anterior muscle. Fluorescein isothiocyanate (FITC)-labeled dextran (MW 150 kDa) was used for an in vivo fluorescent plasma marker of the muscle capillaries. Following a bolus injection of FITC-dextran solution into the caudal vein, the peak blood velocity in the muscle capillaries was measured prior to, during, and following exposure to static magnetic fields (SMF) or 50-Hz electromagnetic fields (EMF) using a fluorescence epi-illumination system. The whole body of experimental animals, placed on the observing stage of a fluorescence microscope, was exposed to SMF (0.3, 1 and 10 mT) or 50-Hz EMF (0.3 and 1 mT) for 10 min using a specially devised electromagnet. For sham exposure, the electromagnet was not energized. During exposure and post-exposure to SMF of 10 mT, the peak blood velocity significantly increased as compared to sham exposure. After the withdrawal of SMF and 50-Hz EMF of 1 mT, significant similar effects on the blood velocity were present or enhanced. These findings suggest that field intensity of 1 mT might be considered as a threshold level for enhancing muscle microcirculation under pentobarbital-induced hypnosis.

Animals↗

PubMed2001

Migration and metalloproteinases determine the invasive potential of mouse melanoma cells, but not melanin and telomerase.

The biological characteristic cell locomotion and invasion, melanin content, metalloproteinases and telomerase activity were studied in a parental mouse melanoma cell line B16 and two descendents B16BL6 and B16F10. The invasive potential of melanoma cells was assayed in a transwell cell culture chamber. Melanin content was determined by the absorbance value at 470 nm per 10(6) cells. Tumor cells migration within the 3-D collagen matrix was microscopically recorded with a time-lapse video recorder and analyzed by computer-assisted cell tracking. Gelatin zymography was adopted to assay the metalloproteinases secretion. A polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) was used for measuring telomerase activity. The results demonstrated that B16BL6 and B16F10 cells were highly invasive compared to B16 cells, but the melanin content of B16F10 was very low. B16F10 and B16BL6 were hypermotile and secreted much more metalloproteinases than B16. No differences were observed in telomerase activity among the three melanoma cell lines. Invasion of mouse melanoma was closely correlated to tumor cell migration and secretion of metalloproteinases. Melanin content and telomerase activity were phenotypically not related to invasiveness in these three mouse melanoma cell lines.

Animals↗

PubMed2001

Functional importance and caffeine sensitivity of ryanodine receptors in primary lymphocytes.

Calcium signaling patterns are important for the specific regulation of activation and effector function in lymphocytes. Studies of [Ca2+]i regulation in lymphocytes, including the involvement of ryanodine receptors (RyR) and the importance of caffeine-sensitive pools, have been carried out mainly in lymphocyte cell lines and the presence and functional importance of these pools in primary lymphocytes has not been addressed. Here we show by confocal microscopy that caffeine caused a prompt but transitory increase of [Ca2+]i in primary lymphocytes, an effect that was inhibited by pre-treatment with ryanodine. Furthermore, the increase of [Ca2+]i in CD4+ and CD8+ MLR T lymphocytes stimulated by 5 microg/ml concanavalin A was significantly inhibited by pretreatment with caffeine. In functional studies, caffeine decreased cytotoxicity against myocyte target cells which is probably related to an altered calcium signaling in CD8+ MLR lymphocytes. Caffeine also terminated spontaneous Ca2+ oscillations and induced a rise in [Ca2+]i in CD4- and CD8- MLR lymphocytes probably of B cell origin. These results demonstrate that caffeine alters Ca2+ signaling in primary lymphocytes, and suggest that RyR, probably the skeletal muscle receptor (RyR-1) and brain receptor (RyR-3), are involved in mediating this effect. It is also possible that blocking of inositol-1,4,5-triphosphate (IP3) receptors is involved in the effects of caffeine on lymphocyte activation.

Animals↗

PubMed2001

Conformational changes and some functional characteristics of gelatin esterified with fatty acid.

This study investigated the effects of attachment of fatty acid chains to gelatin molecules on their conformation and some functional properties in order to determine the effectiveness of this procedure for improving the emulsifying properties of gelatin. The esterification conferred upon the gelatin molecules a folded configuration resulting in a lambda(max) of fluorescence emission shift to lower wavelengths and decreases in solubility, intrinsic viscosity, and gelling ability. In contrast, surface hydrophobicity increased with extent of esterification, and the esterified derivatives exhibited higher emulsion stabilities. These results indicated that the oligomeric structure and functional attributes of the gelatin were altered with the modification.

Emulsions↗

PubMed2001

Interleukin-6 (IL6) and cellular response to facial nerve injury: effects on lymphocyte recruitment, early microglial activation and axonal outgrowth in IL6-deficient mice.

Nerve injury triggers numerous changes in the injured neurons and surrounding non-neuronal cells. Of particular interest are molecular signals that play a role in the overall orchestration of this multifaceted cellular response. Here we investigated the function of interleukin-6 (IL6), a multifunctional neurotrophin and cytokine rapidly expressed in the injured nervous system, using the facial axotomy model in IL6-deficient mice and wild-type controls. Transgenic deletion of IL6 caused a massive decrease in the recruitment of CD3-positive T-lymphocytes and early microglial activation during the first 4 days after injury in the axotomized facial nucleus. This was accompanied by a more moderate reduction in peripheral regeneration at day 4, lymphocyte recruitment (day 14) and enhanced perikaryal sprouting (day 14). Motoneuron cell death, phagocytosis by microglial cells and recruitment of granulocytes and macrophages into injured peripheral nerve were not affected. In summary, IL6 lead to a variety of effects on the cellular response to neural trauma. However, the particularly strong actions on lymphocytes and microglia suggest that this cytokine plays a central role in the initiation of immune surveillance in the injured central nervous system.

Animals↗

PubMed2001

Genetic control of the rate of wound healing in mice.

There have been few studies of the inheritance of wound healing in mammals. In this study, we demonstrate that inbred strains of mice differ significantly in the rate of wound healing. Of the 20 strains tested, fast healers (MRL/MpJ-Fas(lpr) and LG/J) healed wounds four times faster than slow healers (Balb/cByJ and SJL/J). The genetic basis underlying the difference in the healing capacity was analysed using F2 populations of two different crosses. We show that the wound healing is a polygenically determined quantitative trait with an average estimated heritability of 86%. The modes of gene action in these two crosses are different. In the (MRL/MpJ x SJL/J) cross, genes regulating fast healing in MRL/MpJ mice exhibited additive effects, whereas these effects were suppressed by a dominant repressor gene in CBA/J mice in the (MRL/MpJ-Fas(lpr) x CBA/J) cross. Information gained from this investigation provides insight into further study of molecular mechanisms underlying the rate of wound healing in mammals.

Analysis of Variance↗

PubMed2001

EPR investigation of the free radicals generated during the photosensitization of TiO2 colloid by hypocrellin B.

The cation radical of dye produced from the interfacial electron transfer from a surface chelated dye to the conduction band of the colloidal TiO2 was studied by laser flash photolysis and electron paramagnetic resonance (EPR) techniques. The study employed hypocrellin B (HB), a natural photodynamic pigment with strong absorption over the visible light region, as a sensitizer and titanium dioxide as a colloid semiconductor. HB formed a chelate with this colloid semiconductor and exhibited a red-shifted and strongly enhanced absorption in the visible spectrum. Laser photolysis indicated that the electron excitation in the visible absorption band of the chelate resulted in extremely rapid and efficient electron injection from the excited triplet state of the dye into the conduction band of the semiconductor. A transient absorption of cation radical of HB at 570 nm was observed. The appearance of cation radical of HB was characterized by EPR spectrometry: the photoinduced EPR signal was not quenched by oxygen and its intensity decreased in the presence of NaI, a typical hole scavenger. The generation of conduction band electrons in HB-sensitized TiO2 system was also verified by the spin elimination of a stable cyclic nitroxide, 2,2, 6,6-tetramethylpiperidine-1-oxyl (TEMPO), and by the reduction of methyl viologen (MV2+) to its radical MV+.

Animals↗

PubMed2001

Chelation of hypocrellin B with zinc ions with electron paramagnetic resonance (EPR) evidence of the photodynamic activity of the resulting chelate.

Hypocrellin B (HB), a perylenequinone derivative, is an efficient phototherapeutic agent. The chelation of HB with Zinc ions (Zn2+) results in a metal chelate (Zn-HB) which exhibits considerable absorption (lambda max = 612 nm) in the phototherapeutic window. The structure of this chelate has been characterized by UV-Vis, IR and mass spectra. The redox potentials of the Zn-HB chelate were Eox = +1.1 V (vs. SCE) and Ere = -0.7 V (vs. SCE) as measured using the circle volt curve. The quantum yield of singlet oxygen generated by the Zn-HB chelate was 0.86, which both the electron spin trap (EPR) method and the chemical trap method show to be about 0.1 higher than that of its parent compound HB. In irradiated oxygen-saturated solutions of Zn-HB chelate, superoxide radical anions and hydroxyl radicals were detected by EPR spectroscopy using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as the spin-trapping agent.

Chelating Agents↗

PubMed2001

Gelatin: a valuable protein for food and pharmaceutical industries: review.

Many works have appeared in various scientifically reputable journals and publications worldwide that seem to have made potential or satisfactory contribution to our knowledge on the functions and utilization of gelatin--an important source of animal protein. Irrespective of these worldwide publications, room still exists for more work to be done to fully understand the utilization, chemical, biological, physical and functional properties of gelatin. Chemical and enzymatic modifications as well as biological studies should be undertaken with accuracy to be able to extend the utilization of gelatin in food and pharmaceuticals.

Chemistry, Pharmaceutical↗