Biomedical subjects

X Shao

Publications and source records attributed to X Shao.

At least 19 recordsLinked to original sources

PubMed2004

Efficient inhibition of multidrug-resistant human tumors with a recombinant bispecific anti-P-glycoprotein x anti-CD3 diabody.

Overexpressing of P-glycoprotein (Pgp) has been shown to be responsible for cancer resistance to multiple chemotherapeutic agents. Immunotherapy with biological agents, such as bispecific antibodies (BsAbs), may represent a promising approach to overcome the emergence of drug resistance. Here we constructed a recombinant BsAb, a diabody, with specificities to both CD3 on human T-lymphocyte and Pgp on cancer cells. The diabody was produced in Escherichia coli in a soluble functional form and purified by an affinity chromatography with a yield of >4 mg/l culture medium in shaker flask. The diabody binds to both CD3 on T-lymphocytes and Pgp on multidrug-resistant (MDR) tumor cells with affinities that are comparable to its respective parental single chain Fv molecules. In the presence of activated human peripheral blood lymphocytes (PBLs), the diabody mediates effectively the lysis of the Pgp-overexpressing human leukemia K562/A02 and epidermoid carcinoma KBv(200) cells, but is much less potent in mediating the lysis of the parent K562 and KB cells. Further, the diabody localized selectively within the K562/A02 xenografts in mice. When combined with activated PBL, the diabody significantly inhibited the growth of K562/A02 and KBv(200), but had no effect on K562 and KB xenografts. In contrast, treatment with doxorubicin, a standard chemotherapeutic agent, only inhibited the growth of K562 and KB, but had no effect on K562/A02 and KBv(200) xenografts. Taken together, our results suggest that the anti-Pgp x anti-CD3 diabody may have a great potential in the treatment of various MDR cancers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2001

Testicular protein Spag5 has similarity to mitotic spindle protein Deepest and binds outer dense fiber protein Odf1.

Outer dense fibers (ODF) and the fibrous sheath (FS) are major cytoskeletal structures in the mammalian sperm tail. The molecular mechanisms underlying their morphogenesis along the axoneme or their function are poorly understood. Recently, we reported the cloning and characterization of Odf2, a major ODF protein, and Spag4, an axoneme-binding protein, by virtue of their strong interaction with Odf1, the 27 kDa major ODF protein. We proposed a crucial role for leucine zippers in molecular interactions during sperm tail morphogenesis. Here we report the cloning and characterization of a novel gene, Spag5, which encodes a 200 kDa testicular protein that interacts strongly with Odf1. Spag5 is transcribed and translated in pachytene spermatocytes and spermatids. It bears 73% similarity with the mitotic spindle protein Deepest of unknown function. We identified two putative leucine zippers in the C-terminal part of the Spag5 protein, the downstream one of which is involved in interaction with Odf1. Interestingly, these motifs are present in Deepest. These results highlight the importance of the leucine zipper in sperm tail protein interactions. Mol. Reprod. Dev. 59: 410-416, 2001.

Amino Acid Sequence↗

PubMed2001

A two-dimensional immune algorithm for resolution of overlapping two-way chromatograms.

A two-dimensional immune algorithm is proposed for resolving the multicomponent overlapping two-way data matrices. The method is a development of the one-dimensional immune algorithm proposed elsewhere. When the inner product of vectors is expanded to the similar operation on matrices, the 1D immune algorithm can be expanded to the 2D algorithm which is suitable for the analysis of two-way data matrices. Both simulated and experimental two-way data sets were investigated by the method, and the results prove that the 2D immune algorithm is an effective tool for resolving the overlapping two-way signals. The effect of noise on the recoveries is also discussed.

Algorithms↗

PubMed2001

A study on the detection of weak photoacoustic signals by stochastic resonance.

On the basis of stochastic resonance (SR) theory a study of the detection of photoacoustic (PA) signals without a lock-in amplifier is presented. The applicability of the SR method is investigated both with simulated signals calculated by use of a Gaussian model and with experimental PA signals. Signal-to-noise ratio (SNR) is improved substantially by adjusting the properties of a non-linear system to realize SR. By use of this method the lock-in amplifier can be removed from a PA spectrometer.

Journal Article↗

PubMed2001

Signal estimation and denoising using VC-theory.

Signal denoising is closely related to function estimation from noisy samples. The same problem is also addressed in statistics (non-linear regression) and neural network learning. Vapnik-Chervonenkis (VC) theory has recently emerged as a general theory for estimation of dependencies from finite samples. This theory emphasizes model complexity control according to Structural Risk Minimization (SRM) inductive principle, which considers a nested set of models of increasing complexity (called a structure), and then selects an optimal model complexity providing minimum error for future samples. This paper applies the framework of VC-theory to signal estimation/denoising. There are three factors important for accurate signal estimation from finite samples: (1) the type of (orthogonal) basis functions used (i.e. Fourier basis, wavelets etc.). (2) The choice of a structure, i.e. ordering of the basis functions according to their 'importance' for accurate signal estimation. This corresponds to the choice of a 'structure' under SRM formulation. (3) Selecting an optimal number of terms (basis functions) from the ordered sequence of basis functions (2), aka model selection or complexity control (in statistics). We propose a methodology for specifying appropriate orderings (2) and an analytic expression for model selection (3) for signal processing applications. We also present empirical comparisons between the proposed methodology and current state-of-the-art wavelet thresholding methods for univariate signals. These comparisons suggest that the prudent choice of a structure (2) and the use of VC-based model selection (3) are critical for accurate signal estimation with finite samples.

Neural Networks, Computer↗

PubMed2001

A monoclonal antibody specific for human thymidine kinase 1.

Previous research has shown that thymidine kinase 1 (TK1), a nucleotide salvage pathway enzyme, is an accurate prognostic and diagnostic tumor marker. However, the current radioisotope assay for TK1 is cumbersome and has hampered the clinical application of this diagnostic technique in cancer management. To overcome the problems of the current radioisotope assay, we have produced monoclonal antibodies (MAbs) using purified TK1 from Raji cell extract. Production and confirmation of their specificity was confirmed using Western blot, immunohistochemical staining, TK1 activity inhibition assays, and enzyme-linked immunoadsorbent assay (ELISA) techniques. Thus, in the future, these antibodies may aid in the early detection of cancer and more accurate prognosis, as well as allowing for an increased ability to study the function of TK1 in basic cellular processes.

Antibodies, Monoclonal↗

PubMed2001

[The mutation of anti-CD3 antibody (HIT3a) gene and its expression].

OBJECTIVE: To improve the expression of anti-CD(3) single chain Fv (scFv) by site mutation and identify its biological activity. METHODS: Anti-CD(3) scFv gene was mutated by PCR, the target clones were screened by both the fingerprints of DNA restriction endonuclease digestion and Western blot, the antigen-binding activity of scFv was examined by FACS, competitive inhibition was performed with (125)I-labeled HIT3a and the cytotoxic effect mediated by the anti-CD(3) scFv-activated T lymphocytes was analyzed by (51)Cr-released assays. RESULTS: The DNA sequencing showed that the 6th amino acid of the anti-CD(3) antibody (HIT3a) heavy chain gene was mutated from E (GAG) to Q (CAG). The expression of mutated anti-CD(3) scFv (m2) was increased by 100 times higher than that of the parent scFv, and there was no difference in the Jurkat cell (CD(3)(+))-binding activity between the (m2) and parent scFv. The preliminary results of competitive assays showed that m2 could partially block the sites of CD(3)(+) Jurkat cells where the parent antibody bound to. Cytotoxicity assays demonstrated that CD(3)AK cells induced by IL-2 and m2 showed stronger cytotoxic effect than that of LAK cells induced by IL-2 alone in vitro. CONCLUSION: By site mutation, a high expression fragment m2 of anti-CD(3) scFv antibody was obtained. The results of some experiments indicated that m2 could bind to CD(3)(+) Jurkat cells, furthermore, by co-stimulated with IL-2, it could activate peripheral T lymphocytes and induce CD(3)AK cytotoxic effect.

CD3 Complex↗

PubMed2001

[Study of specific targeting cytotoxicity mediated by anti-CD(3)/anti-CD(20) Diabody].

OBJECTIVE: To study the specific targeting cytotoxicity mediated by an anti-CD(3)/anti-CD(20) diabody. METHODS: The diabody was purified by affinity chromatography and identified by Western blot assay, size exclusion chromatography, FACS and rosetting assay; the effect of the anti-CD(3)/anti-CD(20) diabody mediated lysis of CD(20)-expressing tumor cells was assayed by (51)Cr release assay in vitro and by human B cell tumor nude mice xenograft model in vivo. RESULTS: The anti-CD(3)/anti-CD(20) diabody could bind both Jurkat cells (CD(3)(+)) and Daudi cells (CD(20)(+)) and appeared to be potent in targeting activated peripheral blood mononuclear cell (PBMC) to lyse Daudi cells in vitro. Furthermore, it inhibited tumor growth and prolonged the survival of mice bearing xenografted Raji cells. CONCLUSION: The diabody proved to be a potent agent for targeting peripheral blood lymphocytes to lyse CD(20) antigen expressing tumor cells in vitro and in vivo.

Animals↗

PubMed2001

[Influence of the hearing loss level and auditory sensation curve type on click evoked SP/AP amplitude ratio in Menière's disease].

OBJECTIVE: To assess the influence of hearing loss level and auditory sensation curve type on click evoked SP/AP amplitude ratio (negative summating potential/action potential) in the diagnosis of Meniere's disease. METHOD: AP and SP was recorded with tympanic ECochG in normal controls (50 ears), patients with sensorineural hearing loss (SNHL) and patients with Meniere's disease (90 ears) which were further divided based upon the degree of hearing loss and audiometric configuration. SP/AP amplitude ratio was calculated and analyzed. RESULT: AP latency in the three groups had no significant differences. AP amplitude in normal hearing subjects was higher than other two patients groups (P < 0.05). Amplitude of SP of control group was higher than that of SNHL group (P < 0.01), but lower than Meniere's disease group (P < 0.01). SP/AP amplitude ratio in Meniere's disease ears was (48.09 +/- 14.83)%, which was significantly higher than that of SNHL group (20.20 +/- 15.00)% (P < 0.01) and normal group (23.85 +/- 8.05)% (P < 0.01). In Meniere's disease ears, SP/AP amplitude ratio was positively correlated to pure tone thresholds (r = 0.5601 P < 0.01). An elevated SP/AP ratio was found in 73% of the Meniere's ears and 6% of the SNHL ears. SP/AP ratio among the different levels of hearing loss in Meniere's disease ears, of the ears with mild hearing loss was lower than those with moderate (P < 0.01) and severe hearing loss (P < 0.01), but between later groups there was no statistical significant difference (P > 0.05). Positive rate of SP/AP ratio was 63% in mild, 83% in moderate, 73% in severe hearing loss ears respectively. SP/AP amplitude ratio in patients with low tone auditory curve type was lower than that with flat type(P < 0.05), the latter had no significant difference (P > 0.05) compared with high tone type. Positive rate was 67.57% in low tone type, 75% in high tone type and 78% in flat tone type respectively. CONCLUSION: Results suggests the increase of SP/AP ratio was of clinical significance in the diagnosis of Meniere's disease, and was with advancing pure tone thresholds in certain range of hearing loss levels. Click evoked SP/AP amplitude ratio was affected by degree of hearing loss and auditory sensation curve as well, perhaps the influence of the former on SP/AP ratio was considerable significant.

Action Potentials↗

PubMed2000

Solid-supported synthesis of a peptide beta-turn mimetic.

[structure: see text]The solid-supported synthesis of a bicyclic diketopiperazine, a potential peptide beta-turn mimetic, is described. The Ugi reaction between the resin ester of alpha-N-Boc-diaminopropionic acid (an amine input), alpha-bromo acid, aldehyde, and isocyanide is the key step in the proposed protocol.

Chromatography, High Pressure Liquid↗

PubMed2000

Common fold in helix-hairpin-helix proteins.

Helix-hairpin-helix (HhH) is a widespread motif involved in non-sequence-specific DNA binding. The majority of HhH motifs function as DNA-binding modules, however, some of them are used to mediate protein-protein interactions or have acquired enzymatic activity by incorporating catalytic residues (DNA glycosylases). From sequence and structural analysis of HhH-containing proteins we conclude that most HhH motifs are integrated as a part of a five-helical domain, termed (HhH)(2) domain here. It typically consists of two consecutive HhH motifs that are linked by a connector helix and displays pseudo-2-fold symmetry. (HhH)(2) domains show clear structural integrity and a conserved hydrophobic core composed of seven residues, one residue from each alpha-helix and each hairpin, and deserves recognition as a distinct protein fold. In addition to known HhH in the structures of RuvA, RadA, MutY and DNA-polymerases, we have detected new HhH motifs in sterile alpha motif and barrier-to-autointegration factor domains, the alpha-subunit of Escherichia coli RNA-polymerase, DNA-helicase PcrA and DNA glycosylases. Statistically significant sequence similarity of HhH motifs and pronounced structural conservation argue for homology between (HhH)(2) domains in different protein families. Our analysis helps to clarify how non-symmetric protein motifs bind to the double helix of DNA through the formation of a pseudo-2-fold symmetric (HhH)(2) functional unit.

Amino Acid Sequence↗

PubMed2000

Roles of the histone H2A-H2B dimers and the (H3-H4)(2) tetramer in nucleosome remodeling by the SWI-SNF complex.

SWI-SNF is an ATP-dependent chromatin remodeling complex required for expression of a number of yeast genes. Previous studies have suggested that SWI-SNF action may remove or rearrange the histone H2A-H2B dimers or induce a novel alteration in the histone octamer. Here, we have directly tested these and other models by quantifying the remodeling activity of SWI-SNF on arrays of (H3-H4)(2) tetramers, on nucleosomal arrays reconstituted with disulfide-linked histone H3, and on arrays reconstituted with histone H3 derivatives site-specifically modified at residue 110 with the fluorescent probe acetylethylenediamine-(1,5)-naphthol sulfonate. We find that SWI-SNF can remodel (H3-H4)(2) tetramers, although tetramers are poor substrates for SWI-SNF remodeling compared with nucleosomal arrays. SWI-SNF can also remodel nucleosomal arrays that harbor disulfide-linked (H3-H4)(2) tetramers, indicating that SWI-SNF action does not involve an obligatory disruption of the tetramer. Finally, we find that although the fluorescence emission intensity of acetylethylenediamine-(1,5)-naphthol sulfonate-modified histone H3 is sensitive to octamer structure, SWI-SNF action does not alter fluorescence emission intensity. These data suggest that perturbation of the histone octamer is not a requirement or a consequence of ATP-dependent nucleosome remodeling by SWI-SNF.

Adenosine Triphosphatases↗

PubMed2000

Reversed-phase liquid chromatography of proteins and peptides using multimodal copolymer-encapsulated silica.

Multimodal copolymer-encapsulated particles for liquid chromatography were prepared by bonding 1-octadecene and unsaturated carboxylic acids on silica particles (5 microm diameter, 300 A pores) for liquid chromatography of proteins. These multimodal copolymer-encapsulated particles can provide both hydrophobic and hydrogen bonding interactions with polar compounds. The chromatographic performance of these multimodal copolymer-encapsulated particles for peptide and protein separations was evaluated under reversed-phase conditions. Compared with typical C8-bonded silica, polymer-encapsulated particles were more stable in acidic mobile phases and provided better recoveries, especially for large proteins (Mr>0.5 x 10(6)). Totally hydrophobic polymer-encapsulated particles were found to produce broad peaks for proteins, and significant improvements were observed by introducing hydrophilic groups (-COOH) onto the polymer-encapsulated surface to form a multimodal phase. For the reversed-phase liquid chromatography of peptides and proteins, improved selectivity and increased solute retention were found using the multimodal polymer-encapsulated particles. More peaks were resolved for the separation of complex peptide mixtures such as protein digests using the multimodal polymer-encapsulated particles as compared to totally hydrophobic polymer-encapsulated particles.

Chromatography, High Pressure Liquid↗

PubMed2000

A novel method to calculate the approximate derivative photoacoustic spectrum using continuous wavelet transform.

A novel method based on continuous wavelet transform (CWT) using Haar wavelet function for approximate derivative calculation of analytical signals is proposed and successfully used in processing the photoacoustic signal. An approximate nth derivative of an analytical signal can be obtained by applying n times of the wavelet transform to the signal. The results obtained from four other different methods--the conventional numerical differentiation, the Fourier transform method, the Savitzky-Golay method, and the discrete wavelet transform (DWT) method--were compared with the proposed CWT method; it was demonstrated that all the results are almost the same for signals without noise, but the proposed CWT method is superior to the former four methods for noisy signals. The approximate first and second derivative of the photoacoustic spectrum of Pr(Gly)3Cl3.3H2O and PrCl3.6H2O were obtained using the proposed CWT method; the results are satisfactory.

Journal Article↗

PubMed2000

An immune algorithm for resolution of multicomponent overlapping chromatograms.

A novel immune algorithm for resolution and quantitative determination of the components in overlapping chromatograms was proposed by imitating biological immune systems. The algorithm takes an overlapping chromatogram as its input and subtracts the chromatograms of standard samples from the input by iteration of a network. When the residual does not change, the network will converge and chromatographic information of the components in overlapping chromatogram will be obtained. Both simulated and experimental data sets were investigated by the method. Results showed that both resolved results and recoveries of quantitative determination are satisfactory. Comparing with conventional least-square method, the immune algorithm is fast in calculation.

Algorithms↗

PubMed2000

Indole-3-glycerol phosphate, a branchpoint of indole-3-acetic acid biosynthesis from the tryptophan biosynthetic pathway in Arabidopsis thaliana.

The phytohormone indole-3-acetic acid (IAA) plays a vital role in plant growth and development as a regulator of numerous biological processes. Its biosynthetic pathways have been studied for decades. Recent genetic and in vitro labeling evidence indicates that IAA in Arabidopsis thaliana and other plants is primarily synthesized from a precursor that is an intermediate in the tryptophan (Trp) biosynthetic pathway. To determine which intermediate(s) acts as the possible branchpoint for the Trp-independent IAA biosynthesis in plants, we took an in vivo approach by generating antisense indole-3-glycerol phosphate synthase (IGS) RNA transgenic plants and using available Arabidopsis Trp biosynthetic pathway mutants trp2-1 and trp3-1. Antisense transgenic plants display some auxin deficient-like phenotypes including small rosettes and reduced fertility. Protein gel blot analysis indicated that IGS expression was greatly reduced in the antisense lines. Quantitative analyses of IAA and Trp content in antisense IGS transgenic plants and Trp biosynthetic mutants revealed striking differences. Compared with wild-type plants, the Trp content in all the transgenic and mutant plants decreased significantly. However, total IAA levels were significantly decreased in antisense IGS transgenic plants, but remarkably increased in trp3-1 and trp2-1 plants. These results suggest that indole-3-glycerol phosphate (IGP) in the Arabidopsis Trp biosynthetic pathway serves as a branchpoint compound in the Trp-independent IAA de novo biosynthetic pathway.

Arabidopsis↗

PubMed2000

Bisulfite-containing propofol: is it a cost-effective alternative to Diprivan for induction of anesthesia?

UNLABELLED: Propofol (Diprivan(TM); AstraZeneca, Wilmington, DE) is a commonly used drug for the induction of general anesthesia in the ambulatory setting. With the availability of a new bisulfite-containing generic formulation of propofol, questions have arisen regarding its cost effectiveness and safety compared with Diprivan(TM). Two hundred healthy outpatients were randomly assigned, according to a double-blinded protocol, to receive either Diprivan(TM) or bisulfite-containing propofol 1.5 mg/kg IV as part of a standardized induction sequence. Maintenance of anesthesia consisted of either desflurane (4%-8% end-tidal) or sevoflurane (1%-2% end-tidal) in combination with a remifentanil infusion (0.125 microg x kg(-1) x min(-1) IV). Patient assessments included pain on injection, induction time, hemodynamic and bispectral electroencephalographic changes during induction, emergence time, and incidence of postoperative nausea and vomiting. The two propofol groups were comparable demographically, and the induction times and bispectral index values during the induction were also similar. However, the bisulfite-containing formulation was associated with less severe pain on injection (5% vs 11%), with fewer patients recalling pain on injection after surgery (38% vs. 51%, P<0.05). None of the patients manifested allergic-type reactions after the induction of anesthesia. The acquisition cost (average wholesale price in US dollars) of a 20-mL ampoule of Diprivan(TM) was $15 compared with $13 for the bisulfite-containing propofol formulation. Therefore, we concluded that the bisulfite-containing formulation of propofol is a cost-effective alternative to Diprivan(TM) for the induction of outpatient anesthesia. IMPLICATIONS: Bisulfite-containing propofol and Diprivan(TM) (AstraZeneca, Wilmington, DE) were similar with respect to their induction characteristics; however, the generic formulation was associated with a smaller incidence of injection pain. Assuming that the drug costs are similar, these data suggest that the bisulfite-containing formulation of propofol is a cost-effective alternative to Diprivan(TM).

Ambulatory Care↗

PubMed2000

Measuring the VC-dimension using optimized experimental design.

VC-dimension is the measure of model complexity (capacity) used in VC-theory. The knowledge of the VC-dimension of an estimator is necessary for rigorous complexity control using analytic VC generalization bounds. Unfortunately, it is not possible to obtain the analytic estimates of the VC-dimension in most cases. Hence, a recent proposal is to measure the VC-dimension of an estimator experimentally by fitting the theoretical formula to a set of experimental measurements of the frequency of errors on artificially generated data sets of varying sizes (Vapnik, Levin, & Le Cun, 1994). However, it may be difficult to obtain an accurate estimate of the VC-dimension due to the variability of random samples in the experimental procedure proposed by Vapnik et al. (1994). We address this problem by proposing an improved design procedure for specifying the measurement points (i.e., the sample size and the number of repeated experiments at a given sample size). Our approach leads to a nonuniform design structure as opposed to the uniform design structure used in the original article (Vapnik et al., 1994). Our simulation results show that the proposed optimized design structure leads to a more accurate estimation of the VC-dimension using the experimental procedure. The results also show that a more accurate estimation of VC-dimension leads to improved complexity control using analytic VC-generalization bounds and, hence, better prediction accuracy.

Algorithms↗