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Biomedical subjects

X Kong

Publications and source records attributed to X Kong.

At least 19 recordsLinked to original sources
PubMed2001

Directed evolution of alpha-aspartyl dipeptidase from Salmonella typhimurium.

Model-free approaches (error-prone PCR to introduce random mutations, DNA shuffling to combine positive mutations, and screening of the resultant mutant libraries) have been used to enhance the catalytic activity and thermostability of alpha-aspartyl dipeptidase from Salmonella typhimurium, which is uniquely able to hydrolyze Asp-X dipeptides (where X is any amino acid) and one tripeptide (Asp-Gly-Gly). Under double selective pressures of activity and thermostability, through two rounds of error-prone PCR and three sequential generations of DNA shuffling, coupled with screening, a mutant pepEM3074 with approximately 47-fold increased enzyme activity compared with its wild-type parent was obtained. Moreover, the stability of pepEM3074 is increased significantly. Three amino acid substitutions (Asn89His, Gln153Glu, and Leu205Arg), two of them are near the active site and substrate binding pocket, were identified by sequencing the genes encoding this evolved enzyme. The mechanism of the enhancement of activity and stability was analyzed in this paper.

Amino Acid Substitution↗
PubMed2001

A new locus for hereditary gingival fibromatosis (GINGF2) maps to 5q13-q22.

Gingival fibromatosis (GINGF) is an oral disorder characterized by enlargement of the gingiva. It occurs either as the sole phenotype or combined with other symptoms. Thus far, one GINGF locus has been mapped on chromosome 2, at 2p21, and a second possible locus has been mapped to 2p13. However, the genes responsible for this disorder have not been elucidated. We identified a four-generation Chinese GINGF family in which the disease manifests within 1 year after birth. After exclusion of the two known GINGF loci in this family, we performed a genome-wide search to map the chromosome location of the responsible gene. We identified a new locus, GINGF2, on chromosome 5q13-q22 with a maximum two-point lod score of 4.31 at D5S1721 (theta = 0.00). Haplotype analysis placed the critical region in the interval defined by D5S1491 and D5S1453. Within this region, calcium/calmodulin-dependent protein kinase IV (CAMK4) is a strong candidate.

Age of Onset↗
PubMed2001

Directed evolution of operon of trehalose-6-phosphate synthase/phosphatase from Escherichia coli.

Trehalose is a nonspecific protective agent for biomacromolecules. Trehalose-6-phosphate synthase (OtsA)/phosphatase (OtsB), which is encoded by the gene operon otsBA located at -42 of the Escherichia coli genome, is the main enzyme system that catalyzes the synthesis of trehalose in E. coli. We cloned the operon and modified it by directed evolution. Unlike in the previously reported work, we modified the whole operon and screened the positive mutant simultaneously. Thus we believe that the gene complex solves the negative effects between two enzymes if one of them diversifies its structure or functions and finds the form most suitable for trehalose synthesis. It thus mimics the natural process, in which the functional improvement of organisms is related to alterations in coordinated enzymes. The evolution procedure was carried out in a sequence of error-prone PCR, shuffling PCR, and then strict screening of the mutants. After screening of a library of more than 4000 colonies, about 15 positive colonies were analyzed, resulting in a higher concentration of trehalose than control. One of them, E. coli TS7, shows 12.3-fold higher trehalose synthesis ability than E. coli DH5alpha. In contrast, we introduced the cDNA sequence of the tps1 gene from Saccharomyces cerevisiae, which has 54% identity with the gene otsA, as one of the templates in shuffling PCR. By hybrid evolution and screening, we obtained 10 positive colonies with higher concentrations of trehalose than control. E. coli TS22 appears to have 5.3-fold higher trehalose synthesis ability than E. coli DH5alpha and 1.6-fold more than E. coli DEF3(pOTS11). This result demonstrated that coevolution and hybrid evolution, as powerful protocols in protein engineering, are effective in modifying enzyme. It indicates that repeating the process of genomic evolution in nature is feasible.

Directed Molecular Evolution↗
PubMed2001

Blocking intercellular adhesion molecule-1 on human epithelial cells decreases respiratory syncytial virus infection.

The respiratory syncytial virus (RSV) causes potentially fatal lower respiratory tract infection in infants. The molecular mechanism of RSV infection is unknown. Our data show that RSV colocalizes with intercellular adhesion molecule-1 (ICAM-1) on the HEp-2 epithelial cell surface. Furthermore, a neutralizing anti-ICAM-1 mAb significantly inhibits RSV infection and infection-induced secretion of proinflammatory chemokine RANTES and mediator ET-1 in HEp-2 cells. Similar decrease in RSV infection is also observed in A549, a type-2 alveolar epithelial cell line, and NHBE, the normal human bronchial epithelial cell line when pretreated with anti-ICAM-1 mAb prior to RSV infection. Incubation of virus with soluble ICAM-1 also significantly decreases RSV infection of epithelial cells. Binding studies using ELISA indicate that RSV binds to ICAM-1, which can be inhibited by an antibody to the fusion F protein and also the recombinant F protein can bind to soluble ICAM-1, suggesting that RSV interaction with ICAM-1 involves the F protein. It is thus concluded that ICAM-1 facilitates RSV entry and infection of human epithelial cells by binding to its F protein, which is important to viral replication and infection and may lend itself as a therapeutic target.

Antibodies, Monoclonal↗
PubMed2001

Sarcolemmal and mitochondrial K(atp)channels mediate cardioprotection in chronically hypoxic hearts.

X. Kong, J. S. Tweddell, G. J. Gross and J. E. Baker. Sarcolemmal and Mitochondrial K(ATP)Channels Mediate Cardioprotection in Chronically Hypoxic Hearts. Journal of Molecular and Cellular Cardiology (2001) 33, 1041-1045. Hypoxia from birth increases the resistance of the isolated neonatal heart to ischemia. We determined if increased resistance to ischemia was due to activation of sarcolemmal or mitochondrial K(ATP)channels. Rabbits (n=8/group) were raised from birth in a normoxic (F(I)O(2)=0.21) or hypoxic (F(I)O(2)=0.12) environment for 8-10 days and the heart perfused with Krebs-Henseleit bicarbonate buffer. A mitochondrial-selective K(ATP)channel blocker 5-hydroxydecanoate (5-HD) (300 micromol/l) or a sarcolemmal-selective K(ATP)channel blocker HMR 1098 (30 micromol/l) were added alone or in combination for 20 min prior to a global ischemic period of 30 min, followed by 35 min reperfusion. Recovery of ventricular developed pressure was higher in chronically hypoxic than normoxic hearts. 5-HD and HMR 1098 partially reduced the cardioprotective effect of chronic hypoxia, but had no effect in normoxic hearts. The combination of 5-HD and HMR 1098 abolished the cardioprotective effect of chronic hypoxia. We conclude that both sarcolemmal and mitochondrial K(ATP)channels contribute to cardioprotection in the chronically hypoxic heart.

Animals↗
PubMed2001

Latency change estimation for evoked potentials: a comparison of algorithms.

Evoked potentials (EPs) have been widely used to quantify neurological system properties. Changes in EP latency may indicate impending neurological dysfunctions. This paper provides a review and a performance comparison of three classes of latency change estimation algorithms: correlation based, adaptive least mean square (LMS) based, and p-norm based algorithms. Data analysis based on computer simulated data and data from impact acceleration and hypoxia experiments were conducted. This concluded that correlation-based algorithms should only be used when the expected latency change is fixed and the noises are not correlated. While all adaptive LMS type algorithms were capable of tracking and estimating latency changes (fixed or variable) under Gaussian noise conditions, the direct LMS-based algorithm reduced the estimation error power given by traditional filter type LMS algorithms as much as 93%. When periodic interference was present, the frequency selective LMS algorithms outperformed other LMS-based algorithms and reduced of the estimation error power by 89%. Alpha-stable noise processes are better approximations of noises found in various EP analysis applications and adaptive p-norm based algorithms are found to be very robust under such noise conditions, eliminate erroneous abrupt latency changes other algorithms would have produced.

Algorithms↗
PubMed2001

Dentinogenesis imperfecta 1 with or without progressive hearing loss is associated with distinct mutations in DSPP.

Dentinogenesis imperfecta 1 (DGI1, MIM 125490) is an autosomal dominant dental disease characterized by abnormal dentin production and mineralization. The DGI1 locus was recently refined to a 2-Mb interval on 4q21 (ref. 1). Here we study three Chinese families carrying DGI1. We find that the affected individuals of two families also presented with progressive sensorineural high-frequency hearing loss (gene DFNA39). We identified three disease-specific mutations within the dentin sialophosphoprotein gene (DSPP) in these three families. We detected a G-->A transition at the donor-splicing site of intron 3 in one family without DFNA39, a mutation predicted to result in the skipping of exon 3. In two other families affected with both DGI1 and DFNA39, however, we identified two independent nucleotide transversions in exons 2 and 3 of DSPP, respectively, that cause missense mutations of two adjacent amino-acid residues in the predicted transmembrane region of the protein. Moreover, transcripts of DSPP previously reported to be expressed specifically in teeth are also detected in the inner ear of mice. We have thus demonstrated for the first time that distinct mutations in DSPP are responsible for the clinical manifestations of DGI1 with or without DFNA39.

Asian People↗
PubMed2001

Watermarking medical signals for telemedicine.

Modern telecommunication infrastructure supports the possibility of delivering quality health care without the physical presence of medical experts. The integrity of biomedical signals being transmitted through communication channels must be established before their utilization. This paper investigates three digital watermarking techniques for signal integrity verification in an electroencephalogram (EEG) monitoring application for brain injury detection. The techniques studied are patchwork, least significant bit, and quantization watermarking methods. The three techniques are evaluated and compared in the following areas: sensitivity to noise contamination, robustness to EEG signal characteristic changes due to brain injury, and consistency under various communication channel models. The patchwork method performs best for noise contamination rejection among the three methods. The noise contamination detection rates of all three methods remain relatively stable across a wide range of EEG characteristics.

Brain Injuries↗
PubMed2001

[Effect of pentoxifylline on promoter activity of human alpha 1(I) procollagen gene].

OBJECTIVE: To study the effect of pentoxifylline (PTX) on promoter activity of human alpha 1(I) procollagen (COL1A1) gene and the influence of PTX on the promoter activity induced by insulin-like growth factor 1 and insulin. METHODS: The constructs of pCOLH2.5 containing -2483 approximately +42bp of the procollagen gene and chloramphenicol acetyltransferase (CAT) as reporter gene were transiently transfected into human skin fibroblasts. The cells were subsequently treated with PTX or IGF-1 or insulin, or PTX plus IGF-1 or insulin. The CAT activity was assessed 24h after PTX and the cytokines added. RESULTS: PTX of 0.4 mmol/L, 2 mmol/L, and 10 mmol/L decreased the CAT activity of pCOLH 2.5 to 82%+/-9%, 58%+/-8%, and 32%+/-13% of the control level, respectively. IGF-1 and insulin increased the activity of the construct. PTX could inhibit the CAT activity of pCOLH2.5 induced by IGF-1 and insulin. CONCLUSIONS: These studies indicate that PTX downregulates the promoter activity of the human COL1A1 gene, while IGF-1 and insulin upregulates it. PTX can inhibit the promoter activity induced by IGF-1 and insulin.

Collagen↗
PubMed2001

[Decomposition of aqueous sodium pentachlorophenolate (PCP-Na) by using TiO2 coating photocatalyst].

Titanium dioxide (TiO2) photocatalysts were coated on hollow glass beads by sodium silicate (Na2SiO3.9H2O) solution. The coated catalysts were characterized by BET, XRD. Their photoactivity were examined using photocatalytic degradation of sodium pentachlorophenolate(PCP-Na) and glucose solution. The impact of catalysts dosage, PCP-Na initial concentration, initial pH and oxygen concentration on reaction kinetics were also studied in detail. The experiments illustrated that the performance of C3-type catalysts was satisfactory. Conditions of the experiments were following: The initial concentration of PCP-Na and CODCr were 10 mg.L-1 and 400 mg.L-1, respectively, and illumination time was 2 hours and catalysts dosage was 2 g.L-1, the removal rates of CODCr and PCP-Na were above 65% and 92% respectively. Optimum dosage of C3-type catalysts was 2-2.5 g.L-1, DO was no less than 3 mg.L-1.

Catalysis↗
PubMed2001

Glycosaminoglycan mimetics: a therapeutic approach to cerebral amyloid angiopathy.

Amyloid deposits characteristic of cerebral amyloid angiopathy lead to vessel rupture and intracerebral hemorrhage. Proteoglycans associate with the amyloid fibril deposits and are thought to play a role in the polymerization of amyloid proteins and the propagation of the deposition process. A series of low molecular weight anionic compounds was developed to mimic the glycosaminoglycan moieties of these proteoglycans. These compounds were tested in different in vitro systems to determine their anti-Abeta amyloid activity. Specific compounds were identified as being anti-fibrillogenic and protective against Abeta-induced cvtotoxicity. Such compounds also did not show intrinsic cellular toxicity could cross the blood-brain barrier (BBB) in vivo, and showed a good safety profile following chronic' exposure. Molecules showing an anti-amyloid profile combined with the ability to cross the BBB represent promising therapeutics for CAA.

Amyloid beta-Peptides↗
PubMed2001

Molecular epidemiology of respiratory syncytial virus.

OBJECTIVE: To determine the epidemiologic pattern of subgroups A and B and genotypes of respiratory syncytial virus (RSV) during two noncontinuous epidemics during 1990-1991 and 1997-1998 in Beijing. METHODS: Nasopharyngeal secretion (NPS) samples of RSV positive or RSV isolates tested by indirect immunofluorescence (IIF) assay were classified into subgroups A and B. Isolates of RSV were divided into at least six different lineages, designated NP1-NP6, by restriction mapping of the N gene. Np1, 3 and 6 were given by subgroup B isolates, while NP2, 4 and 5 were given by subgroup A isolates. Strains of subgroup A were further subdivided into six lineages SHL1-SHL6 on the basis of the SH gene sequence. SH lineages were closely related to each other and to NP1-NP6. Strains of SHL1, 3 and 4 were closely related and belonged to NP2, SHL2 and 6 to NP4, and SHL5 to NP5. RESULTS: Of 145 RSV NPS samples from the 1997-1998 epidemic, 83 (57.2%) were of subgroup B RSV positive, 62 (42.8%) of subgroup A RSV positive. The rate of occurrence of subgroup A to B strains was about 1:1.3. Two of 10 isolates during the epidemic were subgroup A strains, whereas 8 were subgroup B strains. The rate of occurrence of subgroup A to B strains was 1:4. Eight subgroup A strains of 10 isolates from the 1990-1991 epidermic were dominant; the proportion of subgroup A to B strains was 4:1. With 10 RSV isolates in 1997-1998, all 2 subgroup A strains gave N gene fragment restriction pattern NP4, and fell into SH lineage SHL2, whereas 8 subgroup B strains all belonged to NP3. All 8 subgroup A isolates from the 1990-1991 epidemic gave pattern NP4, and fell into SHL2, while 2 subgroup B strains all belonged to NP3. The classification of subgroups A and B deduced from NP patterns corresponded to the definition of these subgroups by monoclonal antibodies. CONCLUSIONS: These observations confirm that subgroups A and B or multiple lineages of RSV co-circulated in Beijing, but different genome types predominated each year. Moreover, very similar viruses were isolated up to more than 5 years ago, indicating that despite apparent diversity of the subgroup A strains, the separate lineages might be relatively stable.

Genotype↗
PubMed2001

[Carcinoma of kidney collecting duct: an analysis of 10 cases].

OBJECTIVE: To study the clinicopathologic characteristics of renal collecting duct carcinoma (CDC). METHODS: A retrospective study was done in 10 cases of CDC. RESULTS: Among 466 cases of renal cell carcinoma admitted in our Institute between January 1989 and June 30, 1999, 10(2.1%) cases of CDC were identified. Seven presented with gross hematuria and 3 with abdominal pain. Radical nephrectomy was done in 9 patients, enucleation of tumor in one. The primary tumor was located predominantly in the renal medulla. Histologic examination showed prominent tubular or tubulopapillary structures. Sarcomatoid carcinoma, cystadenocarcinoma, nests and cords of tumor cells in desmoplastic stroma were identified in some cases. High molecular weight cytokeratin 34 beta E12 was positive in 8 cases and peanut agglutinin in 7 cases. According to Fuhrman's nuclear grade, one was in G2, 4 were in G3 and 5 in G4. Six patients died of metastases within 3 to 23 months (mean 13.3 months), one died of heart disease with tumor free after 19 months, two survived with tumor free for 14 months and 39 months, respectively, one lost from follow-up. CONCLUSION: CDC is a distinctive renal cell carcinoma with prominent clinical appearance and progressive clinical course.

Adult↗
PubMed2000

Near-infrared spectrophotometric determination of tri- and tetrapeptides.

A new method based on the near infrared technique has been developed for the noninvasive and nondestructive determination of the identity and sequences of amino acid residues in small peptides. The method is capable of distinguishing not only peptides with very similar structures (e.g., Gly-Ala-Ala, Gly-Ala-Leu, Leu-Gly-Gly and Gly-Leu-Leu-Gly, Gly-Leu-Gly-Gly, Gly-Gly-Ala-Gly) but also peptides with the same amino acid residues but different sequences (e.g., Gly-Ala-Ala, Ala-Gly-Ala, Ala-Ala-Gly and Gly-Gly-Gly-Ala, Gly-Gly-Ala-Gly).

Peptides↗
PubMed2000

Refinement of the locus for autosomal dominant hereditary gingival fibromatosis (GINGF) to a 3.8-cM region on 2p21.

Hereditary gingival fibromatosis (HGF, MIM 135300; approved gene symbol GINGF) is an oral disease characterized by enlargement of gingiva. Recently, a locus for autosomal dominant HGF has been mapped to an 11-cM region on chromosome 2p21. In the current investigation, we genotyped four Chinese HGF families using polymorphic microsatellite markers on 2p21. The HOMOG test provided evidence for genetic homogeneity, with evidence for linkage in four families (heterogeneity versus homogeneity test HOMOG, chi(2) = 0. 00). A cumulative maximum two-point lod score of 5.04 was produced with marker D2S390 at a recombination frequency of θ = 0 in the four linked families. Haplotype analysis localized the hereditary gingival fibromatosis locus within the region defined by D2S352 and D2S2163. This region overlaps by 3.8 cM with the previously reported HGF region. Single-strand conformation polymorphism and sequence analysis of the coding region of cytochrome P450 1B1 (CYP1B1) excluded it as a likely candidate gene.

Aryl Hydrocarbon Hydroxylases↗
PubMed2000

Sensitivity enhancement of HCACO by using an HMQC magnetization transfer scheme.

Previous theoretical calculations have demonstrated that the multiquantum relaxation rate of (1)H(alpha)-(13)C(alpha)(R(MQ)) is, on average, 1.3 +/- 0.4 or 1.7 +/- 0.6 times slower than the single-quantum relaxation rate of (13)C(alpha)(R(C)) for a sample with or without, respectively, amide protons. By taking advantage of this fact and by using the PEP sensitivity enhancement scheme, an HMQC version of the HCACO experiment has been developed. We demonstrate that this new experiment is 23 and 55% more sensitive than the original HSQC version of the HCACO experiment, at constant times of 7 and 27 ms, respectively, for a sample of the BC domain of the ciliary neurotrophic factor receptor protein dissolved in D(2)O at 20 degrees C.

Algorithms↗
PubMed2000

A J-multiplied HMQC (MJ-HMQC) experiment for measuring (3)J(HNHalpha) coupling constants.

The J-multiplied HSQC experiment (MJ-HSQC: S. Heikkinen et al., J. Magn. Reson 137, 243 (1999)) amplifies J coupling constants m times and allows direct observation of the (3)J(HNHalpha) coupling constants of peptides and proteins (<10 kDa). The drawbacks to this method are line broadening in the f(1)-dimension and lower sensitivity. In the J-multiplied HMQC (MJ-HMQC) experiment described here, the PEP-HSQC pulse sequence is replaced by a sensitivity-enhanced HMQC section, and the total decay time for the J-coupling and the chemical shift evolution is shortened by a period of t(1). This experiment affords narrower linewidth and enhances the sensitivity by 34%, on an average of 105 well-isolated peaks, when compared with the MJ-HSQC experiment.

Algorithms↗