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Biomedical subjects

S Tong

Publications and source records attributed to S Tong.

At least 73 records · Page 4Linked to original sources

Identification and expression of glycine decarboxylase (p120) as a duck hepatitis B virus pre-S envelope-binding protein.

A 120-kilodalton protein (p120) was identified in the duck liver that binds to several truncated versions of duck hepatitis B virus (DHBV) pre-S envelope protein, suggesting p120 may serve as a DHBV co-receptor. The amino acid sequences of tryptic peptides from purified p120 were found to be the duck p protein of the glycine decarboxylase complex (DGD). DGD cDNA cloning revealed extensive protein conservation with the chicken homologue except for several insertions in the N-terminal leader sequence. The DGD cDNA contained no in-frame AUG codon at the predicted initiation site of the open reading frame, and site-directed mutagenesis experiments established an AUU codon as the translational initiator. The DGD protein expressed in rabbit reticulocyte lysates bound truncated DHBV pre-S protein identical to that of p120 derived from duck liver confirming DGD as p120. Moreover, transfection studies in liver- and kidney-derived cells revealed both cell surface and cytoplasmic expression of the protein. Cloning of the glycine decarboxylase cDNA will permit a direct test of whether it functions as a cell surface co-receptor or as a co-factor in the DHBV replication cycles.

Amino Acid Oxidoreductases↗

A short region in the genome of hepatitis B virus is critical for maintenance of high transcript levels.

The majority of hepatitis B virus (HBV) transcripts are not normally spliced during the viral life cycle, but several splice donor and acceptor sites are conserved on HBV transcripts. In particular, the genome region between nt 450 and 500 of the HBV genome appears to be rich in such sequences. In this study we deleted a short 30-nt sequence between a conserved splice donor site at HBV genome position 462 and a splice acceptor site at position 491, thus deleting the surface/polymerase open reading frames by 10 amino acid residues. At the transcriptional level, this deletion led to >99% reduction of the 2.1-kb class of subgenomic transcripts in transfected cells. Nuclear run-on experiments revealed that the transcription rate of the deleted 2.1-kb transcript is unchanged when compared with the wildtype, suggesting a posttranscriptional mechanism for the downregulation of the deleted transcript. In addition, experiments with a replication-competent HBV mutant containing the 30-nt deletion showed that the corresponding 10-amino-acid sequence within the reverse transcriptase domain of the polymerase protein appeared to be nonessential.

Amino Acid Sequence↗

Sociodemographic and behavioural determinants of blood lead concentrations in children aged 11-13 years. The Port Pirie Cohort Study.

OBJECTIVE: To describe the determinants of blood lead concentration in children with long term environmental exposure to lead. DESIGN: Prospective cohort study. SETTING: The lead smelting town of Port Pirie, South Australia, and surrounding townships. PARTICIPANTS: 326 children born in and around Port Pirie, 1979-1982, followed up until age 11-13 years in 1993-1994. MAIN OUTCOME MEASURES: Blood lead concentrations assessed at birth and at multiple ages up to 11-13 years; average lifetime blood lead concentration. RESULTS: Mean blood lead concentration rose sharply over the ages 6 to 15 months, reached a maximum around 2 years of age, and declined steadily as the children grew older. There was no difference in blood lead concentration between boys and girls until they reached the age of 11-13 years, when mean blood lead concentration in boys (8.4 micrograms/dL [0.41 mumol/L]) was slightly higher than in girls (7.5 micrograms/dL [0.36 mumol/L]). Residential area and father's employment site were the two variables most strongly predictive of a child's blood lead concentration at the end of primary school. Poorer-quality home environment was also found to be an independent contributor to blood lead concentrations. CONCLUSIONS: Age-related factors, and possibly recent concerted efforts to decrease entry or re-entrainment of lead into the environment at Port Pirie, have resulted in most children in our study having blood lead concentrations below 10 micrograms/dL (0.48 mumol/L) at the end of their primary school years. Lead exposure during a child's early years remains an important contributor to average lifetime exposure.

Age Factors↗

Nephelometric determination of micro amounts of nucleic acids with protamine sulfate.

Nucleic acids can form large particle complexes with protamine sulfate by electrostatic forces, which results in strong light scattering. Based on this, a nephelometric method is described for sensitive and convenient determination of nucleic acids with protamine sulfate by using a common spectrofluorimeter. Maximum light scattering is produced in the range of pH 2.2-4.4 with the same excitation and emission wavelengths at 365 nm. Under optimal conditions, the calibration curves are linear in the range 0.05-60.0 micrograms cm-3 for nucleic acids. The corresponding detection limits are 12.5 ng cm-3 for calf thymus DNA, 9.0 ng cm-3 for fish sperm DNA, and 18.0 ng cm-3 for yeast RNA, respectively. Six synthetic samples are determined with satisfactory results. The relative standard deviation of five replicate measurements is 3.2% for 2.0 micrograms cm-3 calf thymus DNA.

Animals↗

Hospital admission and mortality differentials of asthma between urban and rural populations in New South Wales.

It remains unclear whether there are any differentials in hospital admission and mortality rates of asthma between urban and rural populations. An observational study was conducted, based on patient hospital records, to examine the distribution of asthma admissions and mortality in New South Wales. Data on all reported cases of asthma were obtained from New South Wales hospitals between 1989 and 1994. Information on deaths of asthma was collected between 1983 and 1992. The hospital admission rates of asthma varied from 4.8 per 1000 in 1990 to 5.4 per 1000 in 1992 for rural population, and from 3.0 per 1000 in 1991 to 3.4 per 1000 in 1992 for urban population. The hospital admission rates were 51.2-69.1% higher for rural residents than urban dwellers. The mortality rates of asthma ranged from 4.8 per 100,000 in 1983 to 8.0 per 100,000 in 1985 for rural population, and from 3.8 per 100,000 in 1983 to 6.0 per 100,000 in 1989 for urban population. The mortality rates of asthma were 3.62-42.85% higher for rural residents than urban dwellers. These results indicate that the non-age-adjusted hospital admission and mortality rates of asthma were considerably higher in rural populations than in urban populations in New South Wales.

Age Distribution↗

Alternative mechanisms of interaction between homotypic and heterotypic parainfluenza virus HN and F proteins.

Cell fusion by human parainfluenza virus (HPIV) type 2 or type 3 requires the coexpression of both the fusion (F) and haemagglutinin-neuraminidase (HN) glycoproteins from the same virus type, indicating that promotion of fusion requires a type-specific interaction between F and HN. In this report we have further investigated the interaction of the ectodomains of the F and HN glycoproteins from HPIV2 and HPIV3. We constructed mutants of the HPIV2 F and HPIV3 F proteins (F'-KDEL) lacking a transmembrane anchor and a cytoplasmic tail, and containing a C-terminal signal for retention in the endoplasmic reticulum (ER). The P12 and P13 F'-KDEL proteins were both found to be retained intracellularly, and neither could induce cell fusion when co-expressed with homotypic HN proteins. Qualitative and quantitative cell-fusion assays also showed that both the P12 F'-KDEL and P13 F'-KDEL proteins have inhibitory effects on P12 F- and HN-induced cell fusion. However, the F-KDEL mutants were found to inhibit cell fusion by two distinct mechanisms. An interaction between P12 F'-KDEL and P12 HN results in intracellular retention of HN, and a block in its transport to the cell surface. In contrast, P13 F'-KDEL was found to suppress the steady-state intracellular expression levels of HPIV2 HN. These results support the conclusion that fusion involves an interaction between the HN and F proteins, and suggest that an association between F and HN may occur in the ER.

Animals↗

Carboxypeptidase D is an avian hepatitis B virus receptor.

The receptor molecules for human and animal hepatitis B viruses have not been defined. Previous studies have described a 170 to 180 kDa molecule (p170 or gp180) that binds in vitro to the pre-S domain of the large envelope protein of duck hepatitis B virus (DHBV); cDNA cloning revealed the binding protein to be duck carboxypeptidase D (DCPD). In the present study, the DCPD cDNA was transfected into several nonpermissive human-, monkey-, and avian species-derived cell lines. Cells transfected with a plasmid encoding the full-length DCPD protein bound DHBV particles, whereas cells expressing truncated versions of DCPD protein that fail to bind the pre-S protein did not. The DHBV binding to DCPD-reconstituted cells was blocked by a monoclonal antibody that neutralizes DHBV infection of primary duck hepatocytes (PDH) and also by a pre-S peptide previously shown to inhibit DHBV infection of PDH. In addition to promoting virus binding, DCPD expression was associated with internalization of viral particles. The entry process was prevented by incubation of reconstituted cells with DHBV at 4 degrees C and by the addition of energy-depleting agents known to block DHBV entry into PDH. These results demonstrated that DCPD is a DHBV receptor. However, the lack of complete viral replication in DCPD-reconstituted cells suggested that additional factors are required for postentry events in immortalized cell lines.

Animals↗

Development of an In Vitro Screening Assay for Compounds that Increase Bone Formation.

Although parathyroid hormone (PTH) has the ability to stimulate bone growth in both rats and humans, its mechanism of action is not fully understood at the molecular level. An in vitro marker that reflects the in vivo anabolic actions of PTH would facilitate the discovery of small-molecule compounds that stimulate bone growth. We therefore compared the patterns of gene expression in three cell lines treated with PTH. The levels of c-fos, collagenase, interleukin-6 (IL-6), and collagen mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR) in three osteoblast-like cell lines. The most responsive marker was c-fos, which was induced 5-10-fold after 1 h of PTH treatment in the UMR106-01 cell line. Because it is a critical early response gene in bone growth, we investigated the possibility of using c-fos stimulation as a method to screen for compounds that can stimulate bone formation. A highly sensitive, medium-throughput RT-PCR assay for c-fos mRNA expression was established using the Taqmantrade mark Detection System (Perkin Elmer, Mississauga, Ontario). Cells were treated with a series of compounds to determine the specificity of c-fos stimulation. Of the compounds tested, only PTH, prostaglandin E(2), 8-bromo-cAMP, and forskolin induced c-fos mRNA levels, indicating that this assay was specific for compounds that are known to induce cAMP and stimulate bone growth. These results indicate that a simple in vitro assay for c-fos may be a reliable method for the screening of compounds that stimulate bone growth in vivo.

Journal Article↗

Regulation of bcl-2 gene family members in human endometrium by antiprogestin administration in vivo.

It is likely that the changes which occur in the endometrium throughout the menstrual cycle involve apoptosis, and that expression of associated genes, such as the bcl-2 family, are regulated by sex steroids. The aim of this study was to investigate the presence of bcl-2, Bax and oestrogen receptor proteins in secretory endometrium collected from ten patients with normal ovulatory cycles 4 or 6 days after the LH surge, and on the same days in a subsequent cycle in which the formation of secretory changes was inhibited by the administration of the antiprogestin mifepristone (RU486) 2 days after the onset of the LH surge. Since some stromal cells display positive immunoreactivity, leucocyte subpopulations of macrophages (CD68-positive) and large granular lymphocytes (CD56-positive) were identified in serial sections. After administration of mifepristone on day 2 after the LH surge, a significant increase in bcl-2 immunoreactivity was observed in glandular and surface epithelium. A positive correlation (0.874) with nuclear oestrogen receptor immunoreactivity in endometrial glands was demonstrated. Subsets of stromal cells, identified as CD68-positive macrophages and CD56-positive large granular lymphocytes displayed positive immunoreactivity for the bcl-2 epitope, which was unaffected by mifepristone administration. Bax immunostaining was similar in control and antiprogestin-treated endometrium. These data indicate that antiprogestin administration inhibits progesterone downregulation of steroid receptors in endometrial glands, resulting in persistence of a proliferative endometrium and accompanying bcl-2 secretion.

Adult↗

Cross-sectional survey of intralymphocytic and serum elements in hypertensive patients.

OBJECTIVE: To investigate the distribution characteristics of some common and trace elements in both serum and lymphocyte in patients with essential hypertension and to analyse their possible role in the pathogenesis of hypertension. METHODS: A cross-sectional study was carried out in 102 hypertensive patients matched with equal number of normotensive subjects by sex, age, and hypertensive family history; all of them were randomly selected from the same community. Both intralymphocytic and serum concentrations of some relevant elements including Na+, K+, Ca2+, Mg2+, Zn2+, Cu2+ and P were measured for all studied subjects, using the inductively coupled plasma spectrometry (ICPS). RESULTS: 1. Both intralymphocytic and serum concentrations of Na+ (P = 0.009, 0.004 respectively), and Ca2+ (P = 0.001, 0.049 respectively) in hypertensive group were significantly higher than those in the normotensives. 2. Stepwise regression analysis disclosed that systolic and diastolic blood pressure had significant correlation with age, serum Na+/K+ ratio and intralymphocytic Ca2+, Mg2+ concentrations (P < 0.001 for both). 3. According to hypertensive family history, the method of multiple variance analysis had been employed for 4 subgroups and the results revealed that the concentrations of serum Ca2+ and lymphocytic Na+ were significantly higher in hypertensives with or without family history than in normotensives without family history. Furthermore, intralymphocytic Na+ concentration was significantly higher in normotensives with positive family history than that in normotensives without family history. CONCLUSION: The serum and intralymphocytic distribution characteristics of elements in hypertensive patients compared with normotensive controls suggested that there might exist certain defects of the ion-transporting system on cell membrane which might be genetically linked.

Adult↗

[A study on relationship between hypertension and polymorphism of ACE gene in male Yi people in Liangshan Yi autonomous prefecture, Sichuan].

OBJECTIVE: To explore relationship between polymorphism of ACE gene and hypertension. METHODS: A cross-sectional study was conducted and PCR technique was used to analyze gene polymorphism. RESULTS: Genotypes DD, ID and II of ACE gene accounted for 13.0% and 9.7%, 50.9% and 48.4%, and 36.1% and 41.9%, respectively, in the normotensives and hypertensives. Frequencies of I and D alleles for ACE gene were 61.6% and 66.1%, and 38.4% and 33.9%, respectively, in the normotensives and hypertensives. There was no significant difference in distribution of II, ID and DD genotypes and I/D allele frequencies of ACE gene between the hypertensives and normotensives in male Yi people. CONCLUSION: Association between polymorphism of ACE gene and hypertension was uncertain.

Adult↗

[Spectrophotometric determination of cinnarizine based on charge-transfer reaction].

The charge-transfer (CT) complex formed between cinnarizine as the donor and 7, 7, 8, 8-Tetracyanoquinodimethane (TCNQ) as the acceptor in acetone-methanol has been studied by spectrophotometric method. Beer's law is obeyed in the range of 0-18 microg x mL(-1) of cinnarizine. The apparent molar absorptivity of CT complex at 743 nm is 1.58 x 10(4) L x mol(-1) x cm(-1). The composition of CT complex is found to be 1 : 1 by Bent-French and Job's methods. The relative standard deviation is less than 3% (n = 10). The method has been applied to the determination of cinnarizine in tablets with satisfactory results.

Calcium Channel Blockers↗

Declining blood lead levels and changes in cognitive function during childhood: the Port Pirie Cohort Study.

CONTEXT: Many studies have found a significant inverse association between early exposure to environmental lead and cognitive function in childhood. Whether these effects are reversible when exposure is reduced is not clear. OBJECTIVE: To assess the reversibility of the apparent effects of lead on cognitive abilities in early childhood by testing whether declines in blood lead concentrations beyond the age of 2 years are associated with improvements in cognition. SETTING: Urban and rural communities surrounding a large lead smelter in Port Pirie, South Australia. PARTICIPANTS: A total of 375 children followed up from birth to the age of 11 to 13 years. DESIGN: Long-term prospective cohort study. MAIN OUTCOME MEASURES: The Bayley Mental Development Index at age 2 years, the McCarthy General Cognitive Index at age 4 years, and IQs from the Wechsler Intelligence Scale (revised version) at ages 7 and 11 to 13 years. RESULTS: Mean blood lead concentrations in the children decreased from 1.02 pmol/L (21 .2 microg/dL) at age 2 years to 0.38 micromol/L (7.9 microg/dL) at age 1 1 to 13 years, but cognitive scores in children whose blood lead concentration declined most were generally not improved relative to the scores of children whose blood lead levels declined least. Changes in IQ and declines in blood lead levels that occurred between the ages of 7 and 11 to 13 years (r= 0.12, P= .09) suggested slightly better cognition among children whose blood lead levels declined most. CONCLUSION: The cognitive deficits associated with exposure to environmental lead in early childhood appear to be only partially reversed by a subsequent decline in blood lead level.

Child↗

Seasonal rainfall variability, the incidence of hemorrhagic fever with renal syndrome, and prediction of the disease in low-lying areas of China.

To investigate determinants of hemorrhagic fever with renal syndrome (HFRS) in low-lying areas of China, the authors studied Chuigang and Wanggang communities in Anhui Province. These adjacent farming communities have a population of about 100,000. Data were collected from the two communities in 1961-1977 and from Yingshang County in 1983-1995; information covered the incidence of HFRS, amount of precipitation, differences in the water level of the Huai River, density of Apodemus agrarius, autumn crop production, and areas of inundated farmland. Correlation and multiple linear regression analyses were used to estimate the relation between seasonal rainfall, density of mice, occupational factors, and occurrence of the disease. Associations were observed between the incidence of HFRS and the amount of precipitation, the water level of the Huai River, and the areas of inundated farmland in Chuigang community. The smaller the water-level difference, the less farmland was inundated and the higher the incidence of HFRS. In Wanggang community, the density of A. agrarius (r1=0.63, p=0.02), the water-level difference in the Huai River (r2=-0.81, p=0.007), and crop production (r3=0.96, p=0.005) were correlated with the incidence of HFRS. The regression analyses based on Wanggang community suggested that these indexes could be used as predictive variables, and the results from the model were well calibrated with the actual incidence of HFRS in that community (R2=0.88, p < 0.01) and Yingshang County (R2=0.91, p < 0.01).

Animals↗

On-line pretreatment and determination of Pb, Cu and Cd at the microgram l-1 level in drinking water by chelation ion chromatography.

A novel, highly sensitive method for the simultaneous separation and determination of lead, copper, cadmium and other transition metals in drinking water was achieved by on-line sample pretreatment of chelation ion chromatography. Manganese, which coeluted with cadmium, was oxidized to permanganate by ammonium persulfate before injection. Permanganate, with bulk quantity of alkali, alkaline earth metals, iron and aluminum, was eliminated by pyrophosphoric acid-ammonium acetate buffer solution (pH 5.5), while retaining heavy and transition metals on a selective chelating resin (MetPac CC-1 column). Then, they were disabsorbed and transferred to a sulfonated cation exchanger (TMC-1 column). Finally, the concentrated trace metals were separated on a bifunctional ion-exchange column (CS5A) by a concentration gradient of oxalic acid and sodium nitrate eluents, coupled with post-column spectrophotometric detection with 2-[(5-bromo-2-pyridyl)azo]-5-diethylaminophenol (5-Br-PADAP) at 560 nm. The separation and color-development conditions were optimized. The detection limits for the method (signal-to-noise ratio = 3:1) were at or below the microgram l-1 level. The results of drinking water analyses were satisfactory.

Cadmium↗

Analysis of linear, area and volume distortion in 3D ultrasound imaging.

We have developed a three-dimensional (3D) ultrasound imaging system that uses a side-firing probe, axially rotated under computer control, to acquire a series of 2D images, from which the 3D image is reconstructed. For an undistorted reconstruction, the inner radius R0 of the 2D images and the total scanning angle theta must be known accurately. Here, we describe (a) a theoretical analysis of the relative distortion in image shape, length, area, and volume due to an error delta R in R0 or delta theta in theta; (b) measurements of these in simulated and real 3D images; and (c) a method to calibrate R0, theta, and image scale accurately. Theoretically, all four relative distortions vary as P delta R/R + Q delta theta/theta, where magnitude of P < or = 1, magnitude of Q < or = 1, and R is the average distance of the object from the axis. In every case, the simple theoretical formulas for P and Q agree with image measurements to within the measurement uncertainty.

Algorithms↗

Intra- and inter-observer variability and reliability of prostate volume measurement via two-dimensional and three-dimensional ultrasound imaging.

We describe the results of a study to evaluate the intra- and inter-observer variability and reliability of prostate volume measurements made from transrectal ultrasound (TRUS) images, using either the (optimal) height-width-length (HWL) method (V = pi/6 HWL) with two-dimensional (2D) TRUS images (obtained as cross-sections of three-dimensional [3D] TRUS images) or manual planimetry of 3D TRUS images (the 3D US method). In this study, eight observers measured 15 prostate images, twice via each method, and an analysis of variance (ANOVA) was performed. This analysis shows that, with the 3D US method, intra-observer prostate volume estimates have 5.1% variability and 99% reliability, and inter-observer estimates have 11.4% variability and 96% reliability. With the HWL method, intra-observer estimates have 15.5% variability and 93% reliability, and inter-observer estimates have 21.9% variability and 87% reliability. Thus, in vivo prostate volume estimates from manual planimetry of 3D TRUS images have much lower variability and higher reliability than HWL estimates from 2D TRUS images.

Analysis of Variance↗