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Biomedical subjects

L Ding

Publications and source records attributed to L Ding.

At least 37 records · Page 2Linked to original sources
PubMed2001

Human catalytic antibodies with glutathione peroxidase activity.

In order to generate catalytic antibodies with glutathione peroxidase (GPx) activity, we prepared GSH-S-DNP butyl ester and GSH-S-DNP benzyl ester as the haptens. Two ScFvs that bound specifically to the haptens were selected from the human phage-displayed antibody library. The two ScFv genes were highly homologous, consisting of 786 bps and belonging to the same VH family-DP25. In the premise of maintaining the amino acid sequence, mutated plasmids were constructed by use of the mutated primers in PCR, and they were over-expressed in E. coli. After the active site serine was converted into selenocysteine with the chemical modifying method, we obtained two human catalytic antibodies with GPx activity of 72.2U/micromol and 28.8U/micromol, respectively. With the aid of computer mimicking, it can be assumed that the antibodies can form dimers and the mutated selenocysteine residue is located in the binding site. Furthermore, the same Ping-Pong mechanism as the natural GPx was observed when the kinetic behavior of the antibody with the higher activity was studied.

Amino Acid Sequence↗
PubMed2001

Atmospheric pressure ion trapping in a tandem FAIMS-FAIMS coupled to a TOFMS: studies with electrospray generated gramicidin S ions.

A tandem FAIMS-FAIMS system for ion trapping at room temperature and atmospheric pressure is described. The first FAIMS device consisted of a side-to-side configuration (sFAIMS) suitable for ion separation, whereas the second FAIMS device was appropriate for ion trapping (tFAIMS). Ions pre-selected by the sFAIMS entered the tFAIMS and were captured by virtual trapping fields at the hemispherical tip of the inner electrode. The use of the sFAIMS, with wider electrode diameters, and consequently better ion separation efficiency than the tFAIMS, lowered the number of background ions captured in the trapping region of tFAIMS, and thus reduced the space charge effects in the trap. This tandem device was coupled to a laboratory built time-of-flight mass spectrometer and was evaluated using the electrospray generated [M + 2H]2+ ion of gramicidin S. The half-time (t1/2) of the exponential decay of the ion cloud in tFAIMS, determined by monitoring the residual intensity of ions extracted from the ion trapping region of tFAIMS after various delay times, was about 2 s.

Algorithms↗
PubMed2001

Inhibition of the function of the FcgammaRIIB by a monoclonal antibody to thymic shared antigen-1, a Ly-6 family antigen.

Thymic shared antigen-1 (TSA-1) is a member of the Ly-6 family of glycosyl-phosphatidylinositol (GPI)-linked proteins. While it has been proposed that TSA-1 may play a role in thymic development, a physiological ligand for this antigen has not been identified. Here we report that a monoclonal antibody (mAb) to TSA-1, generated by immunizing a hamster with CD40 ligand (CD40L)-activated B cells, interferes with the function of FcgammaRIIB on splenic B cells and the B-cell lymphoma cell line, M12, by binding to TSA on the same cells. The interaction of anti-TSA with FcgammaRIIB resulted in an inhibition of the ability of the FcgammaRIIB to cross-link and/or aggregate soluble anti-CD3 or soluble anti-Cbeta T-cell receptor (TCR), leading to an inhibition of induction of expression of CD25 and CD69, interleukin (IL)-2 production and proliferation of naive T cells. Cross-blocking studies with mAbs strongly suggested that a physical association exists between TSA-1 and the FcgammaRIIB on the surface of activated B cells and favour the view that a functional intermolecular association exists between these two distinct membrane antigens.

Animals↗
PubMed2001

Quantifying spatial localization of optical mapping using Monte Carlo simulations.

Optical mapping techniques used to study spatial distributions of cardiac activity can be divided into two categories. 1) Broad-field excitation method, in which hearts stained with voltage or calcium sensitive dyes are illuminated with broad-field excitation light and fluorescence is collected by image or photodiode arrays. 2) Laser scanning method, in which illumination uses a scanning laser and fluorescence is collected with a photomultiplier tube. The spatial localization of the fluorescence signal for these two methods is unknown and may depend upon light absorption and scattering at both excitation and emission wavelengths. We measured the absorption coefficients (micro a), scattering coefficients (micro s), and scattering anisotropy coefficients (g) at representative excitation and emission wavelengths in rabbit heart tissue stained with di-4-ANEPPS or co-stained with both Rh237 and Oregon Green 488 BAPTA 1. Monte Carlo models were then used to simulate absorption and scattering of excitation light and fluorescence emission light for both broad-field and laser methods in three-dimensional tissue. Contributions of local emissions throughout the tissue to fluorescence collected from the tissue surface were determined for both methods. Our results show that spatial localization depends on the light absorption and scattering in tissue and on the optical mapping method that is used. A tissue region larger than the laser beam or collecting area of the array element contributes to the optical recordings.

Animals↗
PubMed2001

Identification and characterization of a spliced C-type lectin-like gene encoded by rat cytomegalovirus.

The English isolate of rat cytomegalovirus (RCMV) encodes a 20-kDa protein with a C-type lectin-like domain that is expressed in the delayed-early and late phases of the viral replication cycle. Genomic sequence analysis of the restriction fragment KpnR of RCMV revealed significant homology to several C-type lectin-containing molecules implicated in natural killer (NK) and T-cell interactions, as well as genes from four poxviruses and African swine fever virus. The gene is spliced into five exons and shows a splicing pattern with exon boundaries similar to those observed in the human differentiation antigen CD69. The cap site of the gene was mapped by RNase protection, 5' rapid amplification of cDNA ends, and primer extension experiments. This analysis demonstrated that the core promoter of the RCMV lectin-like gene contains a GATA rather than a TATA box. Splicing patterns were confirmed with isolates from an infected-cell cDNA library. A unique aspect of the protein is that its translation is not initiated by the canonical methionine but rather by alanine. To study its role in virus replication and pathogenesis, a recombinant virus was constructed in which the gene is interrupted. Replication in tissue culture was similar to that of wild-type virus.

Amino Acid Sequence↗
PubMed2001

Analyses of microbial community within a composter operated using household garbage with special reference to the addition of soybean oil.

A commercially available composter was operated using fixed composition of garbage with or without the addition of soybean oil. The composter was operated without adding seed microorganisms or bulking materials. Microflora within the composter were analyzed by denaturing gradient gel electrophoresis (DGGE) in the case of oil addition, or by 16/18 S rRNA gene sequencing of the isolated microorganisms in the case of no oil addition. The results showed that, irrespective of the addition of oil, the bacteria identified were all gram positive, and that lactobacilli seemed to be the key microorganisms. Based on the results, suitable microflora for use in a household composter are discussed.

Journal Article↗
PubMed2001

New supports for enzyme immobilization based on copolymers of vinylene carbonate and acrylamide.

In this study, a series of beadlike and hydrophilic supports containing reactive cyclic carbonate groups for enzyme immobilization were prepared via reverse-phase suspension copolymerization of the aqueous solutions of vinylene carbonate (VCA), acrylamide (AA), and N,N'-methylene bisacrylamide in paraffin oil. The supports were used as a matrix for immobilization of trypsin and showed a considerable capacity to couple with trypsin and reasonable retention of activity for the immobilized trypsin, depending on the immobilization conditions, such as the content of VCA structural units, reaction time, and pH of the medium.

Acrylamide↗
PubMed2001

[Characteristics of granular sludge during start-up of the internal circulation].

The quick start-up of the laboratory scale IC reactor and the characteristics of granular sludge during start-up were studied in this paper. The results showed that the first start-up of IC reactor could be finished in 20 days, while secondary start-up only needed 15 days with COD loading rate of 12-15 kg.(m3.d)-1 and COD removal larger than 85%. During start-up, the characteristics of granular sludge changed greatly: average granular diameter was increased from 0.88 mm to 1.25 mm; average settling velocity was enhanced from 35.4 m.h-1 to 105.17 m.h-1; methanogenic activities of the granular increased up to 4 times as large as the seeded sludge; the main methanobacteria was changed from Methanothrix to Methanococcus and Methanobacterium.

Sewage↗
PubMed2001

[The effects of the polysaccharides from Dermatocarpon miniatum on oxygen radicals and lipid peroxidation].

OBJECTIVE: To study the effects of Dermatocarpon miniatum (DEM) polysaccharides scavenging oxygen radicals and inhibiting lipid peroxidation. METHODS: .OH was produced by Fenton reaction and O2-. was produced by the oxidation of pyrogallol. The inhibition to LPO was determined by the colorimetry for testing the relative content of MDA. RESULTS: DEM polysaccharides could scavenge oxygen radical. The amounts of scavenging 50% (EC50) .OH was 1.72 mg/ml, the EC50 for O2-. was 2.12 mg/ml. DEM polysaccharides could also decrease the content of MDA. CONCLUSION: DEM polysaccharides has the effects of scavenging oxygen radicals and the inhibition to LPO. Its activities showed positive correlation with the amounts.

Antioxidants↗
PubMed2001

[Evaluation of saliva ferning test in diagnosis of Sjögren's syndrome].

OBJECTIVE: To evaluate saliva ferning test in (SFT) diagnosis of xerostomia in patients with Sjögren's syndrome (SS). METHODS: Dried samples of freshly produced saliva from 78 patients diagnosed as SS according to european community criteria and 80 healthy controls were examined by light microscopy. The crystallization was classified into 4 types according to the ferning phenomenon: uniformity, branching, spreading and integrity (type I normal and type II, III, IV abnormal). Then, the 78 patients underwent labial salivary gland biopsy. According to Tarpley's classification of labial gland biopsy, > or = 2+ was considered positive. RESULTS: (a) The sensitivity and specificity of SFT were 89.74% (70/78) and 83.75% (67/80) respectively. (b) Abnormal SFT was observed in 70/78 (89.74%) of SS and in 13/80 (16.25%) of normal controls. The differences of SFT in SS patients versus normal controls were significant (P < 0.01). (c) The sensitivity of SFT and labial gland biopsy had no significant differences (P > 0.05) as diagnostic tests in SS. CONCLUSIONS: SFT was valuable as a diagnostic test in patients suspective of SS.

Adolescent↗
PubMed2001

[Study on the phase I metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD].

AIM: To study the phase I metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. METHODS: DDPH was administered i.p. to bile duct-cannulated rats. Bile samples were collected before administration and up to 12 h after administration. After being treated with beta-glucuronidase, the bile samples were purified and enriched with C-18 SPE columns, and then were analyzed by LC/DAD/MSD. The samples containing synthesized reference standards of DDPH metabolite 1-(2, 6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethylamino)-propane (M1), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxy-phenylethylamino)-propane (M2), 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3,4- methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3-hydroxy-4- methoxyphenylethylamino)-propane (M4), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2- (3-hydroxy-4-methoxyphenylethylamino)-propane (M5) and 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3-methoxy-4- hydroxyphenylethylamino)-propane (M6) were analyzed by LC/DAD/MSD under identical conditions. RESULTS: The retention times, UV spectra, molecular weights and production spectra (obtained by collision-induced dissociation) of the apparent ions of peak A, B, C, D, E and F in the total ion chromatogram of DDPH treated rat bile sample were consistent with those of M1, M2, M3, M5, M4 and M6, respectively. CONCLUSION: M1, M2, M3, M4, M5 and M6 were identified as the phase I metabolites of DDPH in the rat.

Animals↗
PubMed2001

[An abzyme to catalytize the deiodination of thyroxine].

AIM: To mimic an important family of selenoenzymes in organism-thyroxine (T4) deiodinases and prepare a selenium-containing abzyme catalyzing deiodination of T4. METHODS: A anti-T4 monoclonal antibody was generated by hybridoma methodology and converted into a selenium-containing abzyme by the method of chemical modification. The catalytic activity of the enzyme was measured by RIA method. RESULTS: The abzyme displayed a marked activity of catalyzing deiodination of T4 and a higher specificity to the substrate T4 than that of natural enzyme, and the double reciprocal plots of the initial rates of T3 formation vs. T4 concentration yielded a family of parallel lines. The catalytic activity could be sensitively inhibited by 6-propyl-2-thiouracil (PTU), a competitive inhibitor for dithiothreitol (DTT). CONCLUSION: An abzyme with the diodination activity was first prepared and the reaction mechanism of the enzyme was bisubstrate ping-pong one.

Animals↗
PubMed2001

[Study on the phase II metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD].

AIM: To study the phase II metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. METHODS: DDPH was administered by i.p. to bile duct-cannulated rats. Bile samples were collected before drug administration and up to 12 h after drug administration. After being purified and enriched with C-18 SPE columns the rat bile samples were analyzed by LC/DAD/MSD to identify the peaks of phase II metabolites. The fractions of phase II metabolites were prepared by HPLC and treated with beta-glucuronidase, and then were purified and enriched with C-18 SPE columns and analyzed by LC/DAD/MSD. The corresponding reference standards of DDPH phase I metabolites were analyzed by LC/DAD/MSD under identical conditions. RESULTS: The peaks M7, M8 and M9 in the chromatograms of rat bile samples were the phase II metabolites of DDPH and the enzymatic hydrolysates of M7, M8 and M9 were 1-(2, 6-dimethyl-4-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M2) and 1-(2,6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethyl-amino)-propane (M1) respectively. CONCLUSION: beta-1-O-[3,5-dimethyl-4-[-2-methyl-2-(3,4-dimethoxy-phenylethylamino)- ethoxy]-phenyl]-glucuronic acid (M7, glucuronide of M3), beta-1-O-[2, 4-dimethyl-3-[2-methyl-2-(3, 4-dimethoxy-phenylethylamino)-ethoxy]-phenyl]-glucuronic acid (M8, glucuronide of M2) and beta-1-O-[2-methoxy-4-[1-methyl-2-(2, 6-dimethylphenoxy)-ethylamino-ethyl]-phenyl]-glucuronic acid (M9, glucuronide of M1) were the phase II metabolites of DDPH in rat bile.

Animals↗
PubMed2000

Omega -crystallin of the scallop lens. A dimeric aldehyde dehydrogenase class 1/2 enzyme-crystallin.

While many of the diverse crystallins of the transparent lens of vertebrates are related or identical to metabolic enzymes, much less is known about the lens crystallins of invertebrates. Here we investigate the complex eye of scallops. Electron microscopic inspection revealed that the anterior, single layered corneal epithelium overlying the cellular lens contains a regular array of microvilli that we propose might contribute to its optical properties. The sole crystallin of the scallop eye lens was found to be homologous to Omega-crystallin, a minor crystallin in cephalopods related to aldehyde dehydrogenase (ALDH) class 1/2. Scallop Omega-crystallin (officially designated ALDH1A9) is 55-56% identical to its cephalopod homologues, while it is 67 and 64% identical to human ALDH 2 and 1, respectively, and 61% identical to retinaldehyde dehydrogenase/eta-crystallin of elephant shrews. Like other enzyme-crystallins, scallop Omega-crystallin appears to be present in low amounts in non-ocular tissues. Within the scallop eye, immunofluorescence tests indicated that Omega-crystallin expression is confined to the lens and cornea. Although it has conserved the critical residues required for activity in other ALDHs and appears by homology modeling to have a structure very similar to human ALDH2, scallop Omega-crystallin was enzymatically inactive with diverse substrates and did not bind NAD or NADP. In contrast to mammalian ALDH1 and -2 and other cephalopod Omega-crystallins, which are tetrameric proteins, scallop Omega-crystallin is a dimeric protein. Thus, ALDH is the most diverse lens enzyme-crystallin identified so far, having been used as a lens crystallin in at least two classes of molluscs as well as elephant shrews.

Aldehyde Dehydrogenase↗
PubMed2000

Inhibition of cell migration by 24-kDa fibroblast growth factor-2 is dependent upon the estrogen receptor.

The single-copy gene for fibroblast growth factor-2 (FGF-2) encodes for multiple forms of the protein with molecular masses of 24, 22.5, 22, and 18 kDa. We reported previously that the 24-22-kDa FGF-2 forms inhibit the migration of endothelial and MCF-7 cells by 50% and 70%, respectively. Here we show that this inhibition of migration is mediated by the estrogen receptor (ER). We have found that depletion of the receptor in either cell line abrogates the inhibitory activity of 24-kDa FGF-2 while re-introduction of the ER into deficient cells once again promotes the inhibitory response. To determine whether exposure to 24-kDa FGF-2 resulted in the activation of the estrogen receptor, 3T3 cells were cotransfected with estrogen receptor cDNA and an estrogen regulatory element-luciferase gene reporter construct and treated with 24- and 18-kDa FGF-2. The high molecular weight form stimulated luciferase activity 5-fold while 18-kDa FGF-2 at the same concentration had no effect. Treatment of ER-positive MCF-7 cells transfected with the reporter construct only showed the same results. Inclusion of the pure estrogen antagonist ICI 182,780 blocked the increase in luciferase activity by 24-kDa FGF-2, further indicating that the response was estrogen receptor dependent. Expression of dominant negative FGF receptor 1 inhibited ER activation, indicating that this was the cell surface receptor mediating the effect. Although growth factor-dependent activation of the ER was reported to require mitogen-activated protein kinase-induced phosphorylation at Ser(118) in COS and HeLa cells, this mechanism is not involved with the activation by 24-kDa FGF-2. These results suggest that the addition of 55 amino acids to the amino-terminal end of 18-kDa FGF-2 by alternative translation alters FGF-2 function and allows for the activation of a second signaling pathway involving the estrogen receptor.

3T3 Cells↗
PubMed2000

Concurrent Parkinson tremors.

1. Concurrent resting and postural tremors of patients with idiopathic Parkinson's disease were monitored using transducers responding to angular velocity of rotation. Spectra and correlation functions were calculated for each pair of records. 2. When concurrent tremor spectra share indistinguishable fundamental frequencies, have statistically significant peaks in their coherence spectra at those fundamental frequencies, and show significant peaks in their cross-correlation functions near zero delay, they are classified as linearly dependent. When such tremor records are superimposed, their phase-locked behaviour is evident. 3. Pairs of correlated concurrent tremors, of varying duration, have been observed in both hands, both feet and in either hand and the contralateral or ipsilateral foot. Correlated tremors may be concurrent with other tremors that are independent. We hypothesize that correlated Parkinson tremors arise from one or more common (and possibly unilateral) central sources.

Aged↗
PubMed2000

Association of several small heat-shock proteins with reproductive tissues in the nematode Caenorhabditis elegans.

Immunohistochemical data on 10 of the 14 small heat-shock (smHSPs) proteins in fourth larval stage and adult Caenorhabditis elegans show that the tissues expressing the greatest number of smHSPs are vulva (HSP12s, HSP43 and, under stress, HSP16s) and spermatheca (HSP12s, HSP25, HSP43 and, under stress, HSP16s). HSP43 is also expressed in male tail structures, and following heat-shock HSP16s are expressed in spermatids and spermatozoa.

Aging↗