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Biomedical subjects

K Gu

Publications and source records attributed to K Gu.

At least 19 recordsLinked to original sources
PubMed2005

Excision of a selectable marker in transgenic rice (Oryza sativa L.) using a chemically regulated Cre/loxP system.

Removal of a selectable marker gene from genetically modified (GM) crops alleviates the risk of its release into the environment and hastens the public acceptance of GM crops. Here we report the production of marker-free transgenic rice by using a chemically regulated, Cre/loxP-mediated site-specific DNA recombination in a single transformation. Among 86 independent transgenic lines, ten were found to be marker-free in the T0 generation and an additional 17 lines segregated marker-free transgenic plants in the T1 generation. Molecular and genetic analyses indicated that the DNA recombination and excision in transgenic rice were precise and the marker-free recombinant T-DNA was stable and heritable.

Base Sequence↗
PubMed2004

Effectiveness of nurse counselling in discouraging the use of the infant walkers.

Infant walkers are widely used by caregivers in Singapore despite being recognized as a household hazard. The study determined the effectiveness of nurse counselling in dissuading caregivers from using the walker. Caregivers of children 4 months of age were recruited and divided into the intervention group (nurse's advice and illustrated pamphlets), a conventional group (questionnaire alone) and a control group (without any intervention in separate polyclinic). The percentage of the caregivers, who used the walkers in each group when their child was 9 months old, was taken as a surrogate indicator of effectiveness of nurse's intervention. The study analyzed 708 caregivers. Fewer caregivers (62.7% intervention vs. 80.4% questionnaire alone vs. 83.0% control) used the walker after nurse's advice with illustrated pamphlets. 8% of the users reported walker-related injuries (n=43). Nurses' counselling could be a simple yet effective method to discourage the use of walkers.

Accident Prevention↗
PubMed2003

High-resolution genetic mapping of Xa27(t), a new bacterial blight resistance gene in rice, Oryza sativa L.

Bacterial blight of rice, caused by Xanthomonas oryzae pv. oryzae ( Xoo) (Ishyama) Dye, is one of the serious diseases prevalent throughout Asia. In a previous study, a resistance ( R) locus was transferred from the tetraploid wild rice Oryza minuta to the cultivated rice species, Oryza sativa L. Here, we report the fine genetic mapping of the R locus, tentatively designated as Xa27(t). We performed disease evaluation with an Xa27(t) near-isogenic line, IRBB27, testing 35 Xoo strains collected from 11 countries. The Xa27(t) locus conferred a high level of resistance to 27 strains and moderate resistance to three strains. Resistance of the Xa27(t) gene was developmentally regulated in IRBB27 and showed semi-dominant or a dosage effect in the cv. CO39 genetic background. As a prelude to cloning Xa27(t), a molecular mapping strategy was employed with a large mapping population consisting of 3,875 gametes. Three molecular markers, M336, M1081, and M1059, closely linked to Xa27(t), were identified to facilitate the mapping of Xa27(t) to the long arm of chromosome 6. The Xa27(t) locus was confirmed by chromosome landing of M1081 and M1095 markers on the rice genome. Markers derived from the genomic sequence of O. sativa cv. Nipponbare were used to further saturate the Xa27(t) genomic region. Xa27(t) was finally located within a genetic interval of 0.052 cM, flanked by markers M964 and M1197, and co-segregated with markers M631, M1230, and M449.

Base Sequence↗
PubMed2001

cDNA - protein fusions: covalent protein - gene conjugates for the in vitro selection of peptides and proteins.

We report a method for the synthesis of covalent cDNA-protein fusions for protein display applications. A branched mRNA template was developed which carries a peptidyl acceptor and a reverse transcription primer at the 3'-end. Translation in vitro followed by reverse transcription produced a protein covalently bonded to its encoding cDNA. Both single- and double-stranded cDNA-protein fusions were prepared. cDNA-protein fusions are stable in alkali and resistant to ribonucleases. Their simple preparation and their resistance towards degradation should make cDNA-protein fusions a useful tool for the in vitro selection and evolution of high affinity ligands from large libraries of polypeptides.

Artificial Gene Fusion↗
PubMed2001

Effects of L-arginine cardioplegia on myocardium.

Infusion of L-arginine (a precursor of nitric oxide, NO) in cardioplegia was examined to test its effect on metabolism of myocardium after myocardial ischemia and reperfusion (IR). Twenty-eight patients undergoing valve replacement were involved and randomly divided into two groups: the control group (crystalloid cardioplegia) and the experimental group (crystalloid cardioplegia + L-arginine). Blood samples were taken both before aortic clamping and after aortic unclamping from right radial artery to measure the concentrations of NO2-/NO3-, lactic acid (LA), malondialdehyde (MDA), superoxide dismutase (SOD), and xanthine oxidase (XOD). In the control group, the NO2-/NO3- level decreased at aortic unclamping, and 30 min later, it decreased significantly as compared with that before aortic clamping (p < .05). In the experimental group, it increased at aortic unclamping (p < .05), and 60 min later, increased to the peak. Five, fifteen, and thirty min after aortic unclamping, the concentrations of LA and MDA in the experimental group were lower than those in the control group (p < .05). Thirty and sixty min after aortic unclamping, the concentrations of SOD remained higher in the experimental group than those in the control group (p < .05). There was no difference between groups in the concentrations of XOD. The addition of L-arginine to the cardioplegia can protect the myocardium from injury by releasing nitric oxide.

Aged↗
PubMed2001

The influences of NO and Ach on cGMP levels in two patient populations.

Pulmonary hypertension following cardiac surgery is an important factor affecting postoperative mortality, and its mechanism has not been thoroughly clarified. Cardiopulmonary bypass (CPB) can destroy pulmonary endothelium and aggravate pulmonary hypertension. This study is designed to investigate the impacts of CPB on vascular endothelium-dependent relaxation, and the relations of CPB to pulmonary hypertension. Forty patients undergoing valve surgery were involved. According to their preoperative pulmonary arterial pressure (PAP), these patients were divided into pulmonary hypertension group (H group) and normal group (N group). The concentrations of cyclic guanosine monophosphate (cGMP) were measured at baseline conditions, after acetylcholine (Ach) injection, and during nitric oxide (NO) inhalation. Samples were taken before sternotomy and after weaning from CPB, 4 and 12 hours post-CPB. At baseline, the level of cGMP in the H group was lower than that of the N group by 33.9% before CPB. After initiating the CPB, although the level of cGMP continuously decreased in both groups until weaning from CPB (the N group decreased 33.3%, and the H group decreased 59%). At that point cGMP was higher in N group than in the H group (p < .01). The level of cGMP of both groups tended to recover 4 hours after CPB, but only the N group returned to baseline 12 hours after CPB. After injection of Ach, the level of cGMP of both groups followed the same change as in the baseline, except with different numeric value. The level of cGMP in N group rose ranging from 160.0-197.3%, while it rose ranging from 87.7-168.1% in H group. The levels of cGMP were higher in N group than those in H group at all times following injection of Ach (61.4, 173.3, 202.7, and 188.0%) (p < .01). After inhalation of NO, the level of cGMP of both groups followed the same change as the baseline. The level of cGMP in N group rose ranging from 194.8-320.5%. Although the levels of cGMP were higher in N group than those in H group (6.9, 25.3, 23.3, and 16.6%), significant differences were achieved at the 4 and 12 hour post-CPB periods (p < .05 or p < .01, respectively). It was concluded that the injury of vascular endothelial function caused by CPB was more critical in pulmonary hypertension patients.

Acetylcholine↗
PubMed2000

Psoralen photo-crosslinked mRNA-puromycin conjugates: a novel template for the rapid and facile preparation of mRNA-protein fusions.

We describe the synthesis of a novel type of mRNA template and its use in the preparation of mRNA-protein fusions. A light-induced psoralen crosslinking reaction was used to attach a puromycin-containing oligonucleotide to the 3'-end of an mRNA template. The photo-crosslinked template was found to undergo efficient mRNA-protein fusion formation in rabbit reticulocyte lysate. Fusion formation was subsequently tested with templates carrying puromycin linkers of different length and chemical composition. Short linkers with multiple triethyleneglycol phosphate building blocks allowed the most efficient fusion formation under a wide range of salt conditions. The present method simplifies the preparation of mRNA-protein fusions and thus significantly accelerates the in vitro protein evolution procedure which involves repetitive cycles of fusion production and selection.

Cross-Linking Reagents↗
PubMed2000

Gene therapy-directed osteogenesis: BMP-7-transduced human fibroblasts form bone in vivo.

An ex vivo gene therapy strategy was used to achieve localized skeletal regeneration in vivo. When an adenovirus vector engineered to express bone morphogenetic protein 7 transduced human gingival fibroblasts or rat dermal fibroblasts, these nonosteogenic tissues formed bone and supported the development of hematopoietic tissue when transplanted into immunocompromised mice. Transduced gingival fibroblasts formed marrow-containing ossicles in 100% of transplants after 1-2 weeks in vivo (n = 30). Immunostaining with murine and human-specific antisera raised against osteonectin and in situ hybridization of human-specific Alu genomic sequence demonstrated that the newly formed bone organ was a chimera of both the human donor and the mouse recipient cells. In experiments of greater clinical relevance, AdCMVBMP-7-transduced dermal fibroblasts repaired critical size skeletal defects in rat calvariae (n = 6). The results of this study suggest a bifunctional role of BMP-7-transduced fibroblasts. The transduced, nonosteogenic cells not only secreted biologically active BMP-7 in vitro and in vivo, but also differentiated into bone-forming cells in vivo. This model exploits the use of an easily biopsied, self-regenerating tissue such as gingiva or skin and suggests that local regeneration of tissues by ex vivo gene therapy may not require that autogenous cells be cultured from the tissue that is to be regenerated.

Adenoviridae↗
PubMed2000

Voltammetric determination of mifepristone at a DNA-modified carbon paste electrode.

A new strategy for the preparation of a DNA-modified carbon paste electrode is developed. It is found that the anodic response of mifepristone is greatly enhanced at the dsDNA-modified carbon paste electrode comparing with that obtained at the bare electrode, while the response at a ssDNA-modified electrode is similar to bare electrode. So the dsDNA-modified electrode is employed as a sensitive biosensor for the detection of mifepristone. A linear dependence of the peak currents on the concentration is observed in the range 2.0 x 10(-7) approximately 2.0 x 10(6) mol/L, with a detection limit of 1.0 x 10(-7) mol/L. The relative standard deviation is 4.3% for six successive determinations of 1.0 x 10(6) mol/L mifepristone. The determination of mifepristone tablets is carried out and satisfactory results are obtained.

Biosensing Techniques↗
PubMed2000

Molecular cloning of a human dentin sialophosphoprotein gene.

Dentin sialoprotein (DSP) and dentin phosphoprotein (DPP; phosphophoryn) are two principal dentin-specific non-collagenous proteins. DPP is extremely acidic and is rich in aspartic acid and serine. By virtue of this structure, DPP may bind large amounts of calcium and may facilitate initial mineralization of dentin matrix collagen as well as regulate the size and shape of the crystals. The function of DSP is not known. DSP and DPP are encoded by a single gene in both rat and mouse, and are uniquely expressed in odontoblasts and transiently in pre-ameloblasts. Because DSP and DPP are isolated from dentin as distinct proteins and appear to be present in different amounts, the nascent dentin sialophosphoprotein (DSPP) is likely cleaved to yield DSP and DPP. However, when, where and how the DSPP is cleaved into DSP and DPP is not clear. To further elucidate the structure and function of human DSP and DPP, we have cloned DPP and DSP cDNA by reverse transcriptase-polymerase chain reaction (RT-PCR) strategies, and then cloned and initiated characterization of a human dentin sialophosphoprotein gene. The genomic organization of human DSPP is very similar to that of mouse, containing five exons and four introns, suggesting it is a homologue of mouse dentin sialophosphoprotein (DSPP). Exons 1-4 encode for DSP, while exon 5 encodes for the C-terminus of DSP and the whole DPP. A 4.6-kb RNA transcript was detected on Northern blot analyses of total RNA extracted from immature (open root apices) human teeth using either a human DPP or DSP probe.

Amino Acid Sequence↗
PubMed2000

Treatment of inflamed ferret dental pulps with recombinant bone morphogenetic protein-7.

Recombinant human BMP-7 (bone morphogenetic protein-7, osteogenic protein-1) is osteogenic, dentinogenic and cementogenic when implanted into the appropriate tissue in vivo. However, most studies characterizing the induction of these tissues have implanted BMP-7 into freshly surgerized, clinically healthy tissues. To determine if BMP-7 is dentinogenic in inflamed dental pulps, we applied BMP-7 to inflamed ferret pulps. A single application of 5 microg of a commercial preparation of lipopolysaccharide (LPS) from Salmonella typhimurium directly to the coronal pulp induced a reversible mixed inflammatory exudate of moderate intensity within 3 d. Treatment with a single application of 2.5, 7.5 or 25 microg recombinant human BMP-7/mg collagen (2 mg total mass/tooth) induced reparative dentinogenesis in controls but not LPS treated dental pulps. These data reveal that a single application of up to 50 microg/tooth of exogenous recombinant BMP-7 is insufficient to induce reparative dentinogenesis in ferret teeth with reversible pulpitis. Given that pulp cells in the inflamed tissues likely retain the capacity to respond to exogenous BMP-7, it is possible that insufficient active recombinant protein is available to induce tissue formation in experimentally inflamed dental pulps.

Animals↗
PubMed2000

Isolation and expression in Escherichia coli of cslA and cslB, genes coding for the chondroitin sulfate-degrading enzymes chondroitinase AC and chondroitinase B, respectively, from Flavobacterium heparinum.

In medium supplemented with chondroitin sulfate, Flavobacterium heparinum synthesizes and exports two chondroitinases, chondroitinase AC (chondroitin AC lyase; EC 4.2.2.5) and chondroitinase B (chondroitin B lyase; no EC number), into its periplasmic space. Chondroitinase AC preferentially depolymerizes chondroitin sulfates A and C, whereas chondroitinase B degrades only dermatan sulfate (chondroitin sulfate B). The genes coding for both enzymes were isolated from F. heparinum and designated cslA (chondroitinase AC) and cslB (chondroitinase B). They were found to be separated by 5.5 kb on the chromosome of F. heparinum, transcribed in the same orientation, but not linked to any of the heparinase genes. In addition, the synthesis of both enzymes appeared to be coregulated. The cslA and cslB DNA sequences revealed open reading frames of 2,103 and 1,521 bp coding for peptides of 700 and 506 amino acid residues, respectively. Chondroitinase AC has a signal sequence of 22 residues, while chondroitinase B is composed of 25 residues. The mature forms of chondroitinases AC and B are comprised of 678 and 481 amino acid residues and have calculated molecular masses of 77,169 and 53,563 Da, respectively. Truncated cslA and cslB genes have been used to produce active, mature chondroitinases in the cytoplasm of Escherichia coli. Partially purified recombinant chondroitinases AC and B exhibit specific activities similar to those of chondroitinases AC and B from F. heparinum.

Blotting, Southern↗
PubMed1999

Diabetes and decline in heart disease mortality in US adults.

CONTEXT: Mortality from coronary heart disease has declined substantially in the United States during the past 30 years. However, it is unknown whether patients with diabetes have also experienced a decline in heart disease mortality. OBJECTIVE: To compare adults with diabetes with those without diabetes for time trends in mortality from all causes, heart disease, and ischemic heart disease. DESIGN, SETTING, AND PARTICIPANTS: Representative cohorts of subjects with and without diabetes were derived from the First National Health and Nutrition Examination Survey (NHANES I) conducted between 1971 and 1975 (n = 9639) and the NHANES I Epidemiologic Follow-up Survey conducted between 1982 and 1984 (n = 8463). The cohorts were followed up prospectively for mortality for an average of 8 to 9 years. MAIN OUTCOME MEASURE: Changes in mortality rates per 1000 person-years for all causes, heart disease, and ischemic heart disease for the 1982-1984 cohort compared with the 1971-1975 cohort. RESULTS: For the 2 periods, nondiabetic men experienced a 36.4% decline in age-adjusted heart disease mortality compared with a 13.1% decline for diabetic men. Age-adjusted heart disease mortality declined 27% in nondiabetic women but increased 23% in diabetic women. These patterns were also found for all-cause mortality and ischemic heart disease mortality. CONCLUSIONS: The decline in heart disease mortality in the general US population has been attributed to reduction in cardiovascular risk factors and improvement in treatment of heart disease. The smaller declines in mortality for diabetic subjects in the present study indicate that these changes may have been less effective for people with diabetes, particularly women.

Adult↗
PubMed1999

Transplantation of human pulpal and gingival fibroblasts attached to synthetic scaffolds.

Autologous tissue grafting for the restoration of oral tissues is limited by several factors, including the availability of sufficient donor tissue. One solution to this problem may be to develop substitute tissue grafts by attaching disaggregated autologous cells propagated in vitro to scaffolds composed of natural or synthetic polymers. We have earlier demonstrated that human dental pulp and gingival fibroblasts (HPF, HGF) adhere to non-woven polyglycolic acid (PGA) scaffolds, proliferate and produce extracellular matrix in vitro. We now report that such HPF and HGF adhered to PGA scaffolds survive when implanted into subcutaneous sites in immuno-compromised mice. The transplanted cells synthesize and secrete type I collagen, cellular fibronectin and may express genes implicated in transducing bone morphogenetic protein (BMP) signals. Messenger RNA for BMP-2, -4, -7 (OP-1), the BMP type I receptors Act RI, BMPR-1A and 1B, the type II receptor BMPR-II, and type I collagen were detected by reverse transcription-polymerase chain reaction (RT-PCR). These data revealed that three adult human dental pulp and gingival cell populations, each from individual donors, attached to PGA scaffolds and cultured for 24 h in vitro, survive implantation and express genes indicative of a capacity to produce extracellular matrix. The implanted cells may also express genes associated with responsiveness to BMP-mediated tissue inductive signals.

Adult↗
PubMed1998

Human dentin phosphophoryn nucleotide and amino acid sequence.

Dentin sialoprotein (DSP) and phosphophoryns (DPP) are major dentin-specific non-collagenous proteins and are synthesized by odontoblasts. DPP are extremely acidic, rich in aspartic acid and serine, possess a high affinity for calcium and collagen, and are believed to function in dentin mineralization. Whereas DSP and DPP are the products of a single gene in mouse and rat, an analogous human gene has not been described. Using RT-PCR based cloning strategies, we have cloned human DPP cDNA from immature molar root total RNA. The open reading frame of this human DPP cDNA comprises 2364 bp encoding 788 amino acids rich in serine (58%), aspartic acid (26%) and asparagine (9%). These are mostly arranged as (DSS)n (n = 1-16), DS and NSS motifs. The N-terminal sequence (DDP) matches that obtained from human DPP extracted from the roots of immature teeth. The core protein of this human DPP was calculated to have a molecular weight of 76,906 Da and a net charge of -206 with an isoelectric point of 2.65. Of the serine residues, 53% can potentially be phosphorylated by casein kinases I and II. Thus, this newly cloned human cDNA, which encodes a protein with characteristics similar to rat and mouse DPP, is identified as a human DPP.

Amino Acid Sequence↗
PubMed1998

Antigenic components of Actinobacillus actinomycetemcomitans lipopolysaccharide recognized by sera from patients with localized juvenile periodontitis.

The dominant antigen of Actinobacillus actinomycetemcomitans recognized by high-titer sera from patients with localized juvenile periodontitis is the serotype antigen located in the O-side chains of lipopolysaccharide. Whether such sera contain antibodies reactive with other epitopes in lipopolysaccharide, as is the case for patients with rapidly progressive periodontitis, remains unknown. We prepared and characterized by gas liquid chromatography lipopolysaccharide, lipid A, core carbohydrate with no or few O-side chains (core) and high-molecular-mass carbohydrate-rich in O-side chains (oligosaccharide) from A. actinomyce-temcomitans ATCC 43718 (serotype b, Y4). Using enzyme-linked immunosorbent assay (ELISA), sera from 36 patients with localized juvenile periodontitis were surveyed using whole-cell sonicate as plate antigen. The seven highest titer sera were selected for further study. Specific IgG antibody binding was observed to intact lipopolysaccharide and to all the lipopolysaccharide fractions. The mean titers were highest for intact lipopolysaccharide (138.8 ELISA units), and lipid A (122 ELISA units), followed by the core fraction (81 ELISA units) and the oligosaccharide fraction (69.5 ELISA units). ELISA inhibition revealed that the core fraction at a concentration of 10 micrograms/test well inhibited antibody binding to A. actinomycetemcomitans lipopolysaccharide by a mean value of 56.7%. To further characterize antibody binding to the core fraction, ELISA inhibition was performed using as inhibitor the core carbohydrate fraction of the Re mutant of Salmonella minnesota, which is known to contain only alpha-keto-3-deoxyoctonate residues and phosphate. This fraction at 10 micrograms/test well inhibited binding of antibodies from 6 of 7 test sera with a mean value of 49.2%. Thus, sera from patients with localized juvenile periodontitis contain antibodies that bind to the O-side chains of lipopolysaccharide, as has been previously reported, but they also contain antibodies that bind to lipid A and to lipopolysaccharide core polysaccharide epitopes, specifically to alpha-keto-3-deoxyoctonate moieties. The humoral immune response to A. actinomycetemcomitans in patients with localized juvenile periodontitis is more complex than previously reported and is very similar to that of patients with rapidly progressive periodontitis.

Adolescent↗
PubMed1998

Mortality in adults with and without diabetes in a national cohort of the U.S. population, 1971-1993.

OBJECTIVE: To examine 22-year mortality (1971-1993), causes of death, life expectancy, and survival in a national sample of diabetic and nondiabetic adults according to age, sex, and race. RESEARCH DESIGN AND METHODS: A representative national cohort of 14,374 adults aged 25-74 years was identified in 1971-1975 in the First National Health and Nutrition Examination Survey (NHANES I). Diabetes was ascertained by medical history interview. The cohort was followed for mortality through 1992-1993, with verification of vital status for 96.2% (n = 13,830). Causes of death were determined from death certificates. RESULTS: Diabetic subjects comprised 5.1% of the cohort and accounted for 10.6% of the deaths. Mortality for diabetic subjects increased from 12.4 per 1,000 person-years for those aged 25-44 years at baseline to 89.7 per 1,000 person-years for those aged 65-74 years. The age-adjusted mortality rate was 57% higher for diabetic men than for diabetic women; the rate was 27% higher for diabetic non-Hispanic blacks than for diabetic non-Hispanic whites. Mortality rates were highest for insulin-treated subjects and for those with > or = 15 years' duration of diabetes. Diabetes was listed on the death certificate as the underlying cause of death for only 7.7% of diabetic men and 13.4% of diabetic women. Considering multiple causes of death, heart disease was listed the most frequently and was present on 69.5% of death certificates of people with diabetes. Death rates were higher for diabetic than for nondiabetic subjects in all age, sex, and race groups. The relative risk of death (diabetic versus nondiabetic subjects) declined with age from a value of 3.6 for those aged 25-44 years at baseline to 1.5 for those aged 65-74 years. The relative risk was elevated in diabetic subjects for all major causes of death except malignant neoplasms. Survival of diabetic subjects was lower than that of nondiabetic subjects in all age, sex, and race groups. Median life expectancy was 8 years lower for diabetic adults aged 55-64 years and 4 years lower for those aged 65-74 years. CONCLUSIONS: In this representative national sample of adults, mortality rates were higher for diabetic men than for diabetic women and for diabetic blacks than for diabetic whites. The study confirms the substantially higher risk of death, lower survival, and lower life expectancy of diabetic adults compared with nondiabetic adults.

Adult↗