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S Guan

Publications and source records attributed to S Guan.

At least 37 records · Page 2Linked to original sources
PubMed1999

[Transpetrosal presigmoid approach for removal of giant petroclival tumors].

OBJECTIVE: To discuss the operative technique, advantage and postoperative complications in 41 patients with giant petroclival tumors treated with the supra- and infratentorial transpetrousal presigmoid approach. METHODS: Clinical data were obtained from the review of charts and radiographic images of the 41 patients after operation with temporo-occipotal free osseous flap, partial petrosectomy and mastoidectomy were performed around the labyrinth. The semicircular canals, cochlea and tympanic cavity were protected during the operation. After the super petrous was ligated and cut off, the petroclival region in front of the sigmoid sinus was exposed. RESULTS: In the 41 patients 20 had meningiomas, 20 epidermoid cysts, and one schwannoma. Total resection was performed in 34 patients, subtotal resection in 16, and greater partial resection in one. Major postoperative complications included temporal aphasia (10 patients), cerebral edema (6), brain stem infarct (2), IIIth, VIIth, VIth and posterial group cranial nerves palsies (16, 16, 8, 6 respectively), and cerebrospinal fluid leakage (4 cases). CONCLUSIONS: This approach can expose sufficiently the petroclival region. It can also reveal the area of adjacent sella, interpeduncular and formen magnum, and homolateral III-XIIth cranial nerves. The postoperative complications may be related to operative techniues and tumorous characteristics.

Adolescent↗
PubMed1998

Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system.

Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacteriophage-based site-specific integration system. By cloning an integrase gene (int), an attachment site (attP) and a glucosyl transfer gene cluster from bacteriophage SfX into a suicide vector, and subsequently introducing this construct into S. flexneri SFL124, we obtained a S. flexneri strain (designated SFL1213) expressing the serotype X somatic antigen specificity. The strain retained other characteristics of the parent strain, such as colony shape, growth rate, and Congo red binding property. Stability test showed that the serotype X O-antigen specificity in SFL1213 was 100% stable after being cultured approximately 72 successive hours under non-selective condition. In a mouse pulmonary model, the recombinant strain elicited a significant level of humoral antibodies which recognized the lipopolysaccharide (LPS) of a wild-type S. flexneri serotype X strain. The site-specific insertion system will be useful when stable expression of a cloned single copy gene is desired in the chromosome of S. flexneri vaccine candidate, SFL124.

Animals↗
PubMed1998

Electrospray ionization and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry of permethylated oligosaccharides.

Mass spectra of fragments of permethylated oligosaccharides are analyzed by Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Sustained off-resonance irradiation (SORI) collision-induced dissociation (CID), quadrupolar axialization, multiple stages of isolation and dissociation (MSn), and ion remeasurement are exploited for carbohydrate structural analyses. That SORI CID internal energies are adequate for linkage analysis of a permethylated glucose oligomer is demonstrated by identifying ring-opened fragment ions from MALDI-generated mass-isolated and collisionally activated ions. Ion remeasurement and axialization techniques enhance the sensitivity of ion fragmentation analysis. Multiple stages of isolation and dissociation of ion fragments (MSn) provide for structural analysis of an electrospray-ionized permethylated lacto-N-fucopentaose isomer (LNFP II). Compared to MS2 spectra taken with a triple quadrupole, FT-ICR MSn (n > 2) provides more extensive characterization of the parent molecular structure than is available from a single stage of ion isolation and dissociation (MS2).

Carbohydrate Sequence↗
PubMed1998

N-Glycosylation of pig flavin-containing monooxygenase form 1: determination of the site of protein modification by mass spectrometry.

By using a combination of biochemical methods (i.e., endoglycosidase H digestion and immunoblot and plant lectin binding studies), it was verified that pig flavin-containing monooxygenase (FMO1) was N-glycosylated. By using mass spectrometry approaches [i.e., peptide mapping, gas chromatography/mass spectrometry, microbore HPLC/electrospray ionization mass spectrometry (LC/ESI/MS), chemical ionization gas chromatography/mass spectrometry (CI/GC/MS), and matrix-assisted laser desorption mass spectrometry (MALDI/MS)], we were able to confirm that pig FMO1 was N-glycosylated and we were able to identify the site of N-glycosylation. Pig FMO1 contains two putative consensus sites of N-glycosylation. The results showed that pig FMO1 amino acid Asn120 was selectively N-glycosylated. Highly purified pig FMO1 avidly bound concanavalin A and reacted positively for carbohydrates by the periodic acid/Schiff's base method of analysis. In addition, treatment of pig FMO1 with endo-N-acetylglucosaminidase converted the enzyme to another species with a molecular mass approximately 5000 Da lower than that of the parent protein as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot experiments. Peptide mapping of pig FMO1 showed that the protein used in the study was not contaminated with another glycoprotein. MALDI/MS experiments showed that pig FMO1 was present with the expected molecular mass but that higher-molecular mass forms consistent with the presence of N-linked high-mannose oligosaccharide structures were also covalently attached to the enzyme. The presence of N-acetylglucosamine isolated from acid hydrolysates of the N-linked high-mannose oligosaccharide of pig FMO1 was confirmed by high-pH anion exchange HPLC studies and verified by CI/GC/MS studies of derivatized monosaccharide fractions. Further analysis of pig FMO1 proteolytic peptides by LC/ESI/MS showed that the only residue that was N-glycosylated in pig FMO1 was Asn120. Knowledge of the structural aspects of FMO may be useful in understanding the membrane association properties of the enzyme.

Amino Acid Sequence↗
PubMed1997

A discussion about the DiFrancesco-Noble model.

An obvious defect in the DiFrancesco-Noble (DN) model was found, so methods to overcome this inadequacy are put forward. Based on these methods, different kinds of modified DN model can be obtained, and numerical results show that most of the dynamics of the DN model can be essentially preserved by these modified systems.

Animals↗
PubMed1997

Linear prediction Cholesky decomposition vs Fourier transform spectral analysis for ion cyclotron resonance mass spectrometry.

The fast Fourier transform (FFT) method of spectral analysis converts a time domain signal to a more easily visualized frequency domain spectrum but does not distinguish between signal and noise and produces spectral artifacts (e.g., "Gibb's oscillations") for a truncated and/or improperly sampled time domain signal. For example, FFT cannot resolve two signals if the sampling duration is less than one cycle of the frequency difference between the two signals. Here, linear prediction Cholesky decomposition spectral analysis is applied to ion cyclotron resonance mass spectrometry. The algorithm is robust and capable of extracting spectral parameters (frequency, time domain exponential damping constant, magnitude, and phase) from a signal consisting of multiple exponentially damped noisy sinusoids. Compared to FFT data reduction, linear prediction can offer significantly increased sensitivity (for signals at or below the rms noise level), elimination of Gibb's oscillations, and increased spectral resolving power for a time domain signal that either is truncated or has damped to the rms noise level before the end of the acquisition period. The present analysis can handle up to 8K time domain data sets with 2.5 h PC computation time.

Algorithms↗
PubMed1997

Detection, number, and sequence location of sulfur-containing amino acids and disulfide bridges in peptides by ultrahigh-resolution MALDI FTICR mass spectrometry.

Here, we present several strategies for determining the number of sulfur atoms and disulfide bridges in selected biologically active peptides, based on MALDI FTICR mass spectrometry at femtomole sample consumption level. First, based on the 2-Da mass increase per disulfide bridge reduction, we show that repeated laser shots on the same sample spot can reduce (and therefore reveal the presence of) the disulfide bridge in oxytocin. Second, we show that the primary sequence positions of the disulfide-bridged cystines can be inferred from the presence/absence of MALDI-induced reduction in cystine-containing fragment ions. Third, we show that the presence and number of sulfur atoms as well as the degree of reduction in a peptide can all be determined directly from isotopic relative abundances of mass-resolved 34S, 13C2, and reduced all-12C species in a single ultrahigh-resolution MALDI FTICR mass spectrum. Methods for achieving such ultrahigh mass resolution of peptide ions of closely spaced m/z (m/delta m50% approximately 950,000 at m/z approximately 650) at modest magnetic field (3 T) are discussed.

Amino Acids, Sulfur↗
PubMed1997

Two-way conversation with a mass spectrometer: nondestructive interactive mass spectrometry.

Most mass spectrometers employ destructive detection, so that it is necessary to repeat an experiment in order to vary even one parameter. In contrast, Fourier transform ion cyclotron resonance mass spectrometry offers nondestructive detection, so that ions remain available for further manipulation and redetection. Here, we show for the first time how to perform mass spectrometry interactively. Following each elementary experimental stage, such as ion generation, isolation, dissociation, or detection, the operator is free to choose and tailor the next stage without creating a fresh supply of ions. For example, we can test the effect of varying one parameter over several values without having to repeat the entire experimental event sequence each time, much like varying one letter or word in a sentence without having to rewrite the whole sentence. Such interactive control promises to speed development of complex experimental event sequences, as for optimizing the sequencing and structural analysis of tiny amounts (e.g., femtomoles or less) of biomacromolecules (peptides, nucleic acids, oligosaccharides).

Fourier Analysis↗
PubMed1997

Receptor-mediated endocytosis of immunoglobulin light chains by renal proximal tubule cells.

We examined the binding, endocytosis, and degradation of immunoglobulin light chains by primary cultures from rat renal kidneys and immortalized human proximal tubule cells. Both the association and dissociation of light chain were rapid and plateaued within 30 min at 4 degrees C. Up to 10(-3) M bovine serum albumin did not inhibit light chain binding to cells. Internalization studies with 125I-labeled kappa- and lambda-light chains by cells using the acid wash technique showed that up to 80% of total cell-associated binding at equilibrium (30 min) is rapidly internalized at 22 degrees C. Comparison of binding and internalization of light chains with transferrin, a ligand known to undergo receptor-mediated endocytosis, showed that both ligands displayed saturable kinetics. In contrast, endocytosis of sucrose, a marker for fluid-phase endocytosis, was unsaturable and nearly 200-fold less efficient than light chain internalization. Scatchard analysis of binding experiments done at 4 degrees C with trace 125I-labeled lambda-light chain in presence of 0 to 3.0 x 10(-3) M cold light chain revealed a single class of binding sites with a dissociation constant of 5.0 +/- 0.8 x 10(-5) and a maximal binding capacity of 1.6 +/- 0.3 x 10(-9) mol/mg cell protein. Hypertonic medium, a maneuver which interferes with the formation of the clathrin lattice, reduced endocytosis of light chain significantly but did not affect endocytosis of sucrose. Chloroquine and bafilomycin A, agents that interfere with vesicular acidification, also significantly suppressed light chain endocytosis. Using acid precipitation method, we observed that endocytosis of 125I-labeled lambda-light chain results in degradation by the rat renal proximal tubule cells. Degradation was maximum at 37 degrees C, significantly reduced at 22 degrees C, and absent at 4 degrees C. Excess light chain inhibited degradation of radiolabel, whereas excess albumin had no effect. These studies document the presence of binding sites for light chains on proximal tubule cells that mediate endocytosis of light chains by proximal tubule cells. The present data suggest that receptor-mediated endocytosis of light chains leads to delivery of this ligand to degradative sites through acidified vesicles.

Ammonium Chloride↗
PubMed1996

High-resolution multistage MS, MS2, and MS3 matrix-assisted laser desorption/ionization FT-ICR mass spectra of peptides from a single laser shot.

By combined and repeated use of sustained off-resonance irradiation (SORI) for ion dissociation, stored waveform inverse Fourier transform (SWIFT) waveforms for ion isolation, and ion axialization and remeasurements techniques, we obtain for the first time MS, MS2, and MS3 FT-ICR mass spectra from peptide ions (enzymatic digest products of horse cytochrome c) produced from a single laser shot. The successive fragmentation of gas-phase ions detected from the same initial batch of ions increases the sensitivity of analysis of trace amounts of biological samples in structural mass spectrometry, and fragment identification is facilitated by resolution of carbon-13 isotopic distributions. The method is illustrated by analyses of subfemtomole amounts of crudely purified samples of tryptic digest solutions of horse cytochrome c and bovine cytochrome c. The high-resolution primary ion mass spectrum, along with the collision-induced dissociation (CID) and MSn capabilities of FT-ICR, help to determine the primary amino acid sequence of the fragment ions beyond what is obtained from enzymatic digestion alone, without prior chromatographic separation and purification.

Animals↗
PubMed1996

Electrospray ionization Fourier transform ion cyclotron resonance at 9.4 T.

We present the first results from a new electrospray ionization Fourier transform ion cyclotron resonance mass spectrometer operated at a magnetic field of 9.4 T (i.e. > or = 2.4 T higher than for any prior FTICR instrument). The 9.4 T instrument provides substantially improved performance for large molecules (> or = 50% increase in mass resolving power) and complex mixtures (> or = 100% increase in dynamic range) compared to lower-field (< or = 6 T) instruments. The higher magnetic field makes possible larger trapped-ion population without introduction of significant space--charge effects such as spectral peak shift and/or distortion, and coalescence of closely-spaced resonances. For bovine ubiquitin (8.6 kDa) we observe accurate relative isotopic abundances at a signal-to-noise ratio greater than 1000:1, whereas a complete nozzle-skimmer dissociation electrospray ionization (ESI) FTICR mass spectrum of bovine carbonic anhydrase (29 kDa) is achieved from a single scan with a signal-to-noise ratio of more than 250:1. Finally, we are able to obtain mass resolving power, m/delta m > 200,000, routinely for porcine serum albumin (67 kDa). The present performance guides further modifications of the instrument, which should lead to significant further improvements.

Animals↗
PubMed1996

Fourier transform ion cyclotron resonance mass spectrometry in a 20 T resistive magnet.

We present Fourier transform ion cyclotron resonance (FTICR) mass spectra at a magnetic field of 20 T; more than twice the highest field previously used for FTICR. Our instrument is based on a resistive magnet installed at the National High Magnetic Field Laboratory. The magnet has a 50 mm diameter bore and spatial inhomogeneity of approximately 1000 ppm over a 1 cm diameter spherical volume. However, FTICR mass resolving power far in excess of magnet homogeneity is achieved routinely for ions produced by either electron ionization (EI) or matrix-assisted laser desorption/ionization (MALDI). As examples, we show a MALDI mass spectrum of [M + H] quasimolecular ions of the peptide, human luteinizing hormone-releasing hormone (monoisotopic molecular weight, 1181.6 Da) at mass resolving power, m/delta m > 10,000; and an EI mass spectrum of molecular ions of the platinum cluster compound, Pt4(PF3)8 (average molecular weight, 1484 Da at mass resolving power, m/delta m approximately 20,000. Much better FTICR MS performance is predicted for future NHMFL resistive magnets of higher spatial and temporal homogeneity.

Cyclotrons↗
PubMed1996

A high-performance modular data system for Fourier transform ion cyclotron resonance mass spectrometry.

The three major components of a Fourier-transform ion cyclotron resonance (FT-ICR) mass spectrometer include the vacuum system (including ion source), the magnet and a data system capable of performing the necessary instrument control for desired experiments. Most previous FTICR systems have used commercial data systems based on custom-built electronics controlled by proprietary mini-computers developed in the early 1980's. Here we present a high-performance data system based on a personal computer running user-friendly Windows software and readily available commercial components contained in a VXI chassis and a minimal complement of simple custom electronics. The system uses a VXI pattern generator to control all aspects of the experiment. The flexibility of the pattern generator allows for performance of all current FTICR experimental sequences.

Cyclotrons↗
PubMed1996

Laser-induced fluorescence of Ba+ ions trapped and mass-selected in a Fourier transform ion cyclotron resonance mass spectrometer.

We present the design and preliminary results from a Fourier transform ion cyclotron resonance (ICR) mass spectrometer developed for the direct detection of UV/visible laser-induced fluorescence of trapped, mass-selected, gas-phase ions. A 3 T superconducting magnet and an open-ended multi-section cylindrical Penning trap capture and confine ions created by electron impact or laser desorption. Azimuthal quadrupolar excitation in the presence of ion/neutral collisions cools, axializes and mass selects ions as they fill the trap. A pulsed dye laser pumped by an Nd:YAG laser provides electronic energy excitation. A Brewster window and baffles on each side of the vacuum chamber reduce the scattered light from the excitation laser. Laser-induced fluorescence is collected from mirrors and lenses and directed through a quartz window and fiber-optic bundle to a photomultiplier. The ICR and optical events are controlled by a modular ICR data station and GPIB and RS-232 interfaces. An excitation spectrum is demonstrated for atomic Ba+ ions, and should extend to laser-induced fluorescence of virtually any stable positive or negative gas-phase ions of arbitrary molecular weight: molecular or quasimolecular ions, fragment ions, adduct ions, and ions formed from ion/molecule reactions.

Barium↗
PubMed1996

Determination of ion magnetron radial distribution in Fourier transform ion cyclotron resonance mass spectrometry.

The spatial distribution of the ion cyclotron orbit 'guiding centers' (magnetron radii) in a Penning trap determines virtually all aspects of Fourier transform ion cyclotron resonance (FTICR) mass spectrometry performance. Here, we demonstrate experimentally a simple method for determining that distribution, based on measuring FTICR mass spectral peak height following dipolar excitation/dipolar detection at various stages of magnetron radial expansion. No instrumental modifications are needed. Such data will make it possible to evaluate theoretical models of space charge and to quantitate and optimize ion formation, ion injection, ion trapping, ion transfer, photodissociation and ultrahigh-resolution FTICR experiments.

Cyclotrons↗
PubMed1995

Attomole biomolecule mass analysis by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance.

Significantly improved sensitivity for analysis of biomolecules by MALDI FT-ICR mass spectrometry is achieved by (i) microscope-monitored sample deposition onto a small indentation on the probe tip and (ii) multiple remeasurement of ions from a single laser shot. A simple modification to the solids probe tip allows for microdeposition of a few amols of analyte onto small indentation spots previously aligned with the laser beam. Ion multiple remeasurement of the same ion packet enhances the signal-to-noise ratio and thus extends the achievable FT-ICR MS detection limit. We demonstrate that FT-ICR can be used to detect parent and structurally significant fragment ions of peptides and phospholipids at low amol amounts. Positive ion mass spectra for approximately 90 amol of a mixture of angiotensin II and bradykinin, approximately 40 amol of dipalmitoylglycerophosphatidylcholine, and approximately 8 amol of substance P constitute the lowest reported detection limits to date for FT-ICR mass analysis of MALDI-generated ions.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1995

Phosphate transport inhibition by KW-3902, an adenosine A1 receptor antagonist, is mediated by cyclic adenosine monophosphate.

We have previously demonstrated that 1,3-dipropyl-8-(3-noradamantyl) xanthine (KW-3902) has an inhibitory effect on phosphate (Pi) transport with no effect on glucose transport in the rat renal proximal tubular cell, similar to that of parathyroid hormone (PTH). In the current studies we investigated the effect of KW-3902, rat PTH (1-34), and 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), another selective adenosine A1 receptor antagonist, on Pi transport and the production of cyclic adenosine monophosphate (cAMP). We then compared these effects of KW-3902 with those of rat PTH in rat renal proximal tubule cells. The results showed that both KW-3902 (30 mumol/L) and rat PTH (1-34, 5 mumol/L) significantly inhibited Pi uptake in proximal cells from a control level of 61 +/- 3 to 19 +/- 3 (a reduction of 69%) and 46 +/- 4 picomoles phosphate/mg protein/min (a reduction of 25%), respectively (P < 0.01). The inhibitory effect of 30 mumol/L KW-3902 alone on Pi transport was more than twice that of 5 mumol/L rat PTH (1-34) alone (P < 0.01). KW-3902 stimulated the production of cAMP in a dose-dependent manner (r = 0.997, P < 0.01). Rat PTH (1-34; 5 mumol/L) also stimulated cAMP production, which was greater than that induced by 30 mumol/L KW-3902 alone. A significant increase in cAMP production by 30 mumol/L DPCPX was also observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗