Biomedical subjects

N Chang

Publications and source records attributed to N Chang.

At least 19 recordsLinked to original sources

PubMed2004

Effects of the interaction between the C677T 5,10-methylenetetrahydrofolate reductase polymorphism and serum B vitamins on homocysteine levels in pregnant women.

OBJECTIVE: The purpose of this study was to investigate the effect of the interaction between the C677T mutation in the 5,10-methylenetetrahydrofolate reductase (MTHFR) genotypes and serum levels of B vitamins on serum homocysteine levels in pregnant women. DESIGN: A cross-sectional study. SETTING: Ewha Womans University Hospital, Seoul, Korea. SUBJECTS: A total of 177 normal pregnant women, 24.6+/-1.1 weeks of gestation, in a 6-month period during 2001-2002. INTERVENTIONS: Serum vitamin B2, vitamin B6, and homocysteine analyses were conducted using high-performance liquid chromatography methods. Serum folate and vitamin B12 concentrations were determined using a radioimmunoassay kit. MTHFR gene mutation was investigated by the polymerase chain reaction of a genomic DNA fragment. RESULTS: Serum homocysteine was higher in women with the T/T genotype than those with the C/T or C/C genotype of the MTHFR gene (P<0.05). Serum homocysteine was negatively correlated with serum folate in all MTHFR genotypes (P<0.001), and the correlation between the two serum levels was the strongest in the T/T genotype. Serum homocysteine was higher in the subjects with the T/T MTHFR genotype only when the serum folate was below the median level. Explanatory power of B vitamin status as predictors of serum homocysteine levels was more pronounced in the T/T genotypes (68.5%) compared with the C/T (37.9%) or C/C genotypes (20.6%). CONCLUSIONS: Serum homocysteine levels in pregnant women varied significantly with MTHFR genotype and the serum B vitamin status. Higher serum folate, vitamin B2, and vitamin B12 concentrations may lessen the MTHFR genotypic effect on serum homocysteine levels.

Adult↗

PubMed2001

Development of a Canadian standardized protocol for subtyping methicillin-resistant Staphylococcus aureus using pulsed-field gel electrophoresis.

A panel of 24 methicillin-resistant Staphylococcus aureus strains was distributed to 15 laboratories in Canada to evaluate their in-house pulsed-field gel electrophoresis (PFGE) protocols and interpretation criteria. Attempts to compare fingerprint images using computer-aided analysis were not successful due to variability in individual laboratory PFGE protocols. In addition, individual site interpretation of the fingerprint patterns was inadequate, as 7 of 13 sites (54%) made at least one error in interpreting the fingerprints from the panel. A 2-day standardized PFGE protocol (culture to gel image) was developed and distributed to all of the sites. Each site was requested to use the standardized protocol on five strains from the original panel. Thirteen sites submitted gel images for comparisons. The protocol demonstrated excellent reproducibility and allowed interlaboratory comparisons with Molecular Analyst DST software (Bio-Rad) and 1.5% band tolerance.

Bacterial Typing Techniques↗

PubMed2001

Insulin and IGF-I inhibit calcium-dependent chloride secretion by T84 human colonic epithelial cells.

D-Myo-inositol (3,4,5,6) tetrakisphosphate [Ins(3,4,5,6)P(4)] or phosphatidylinositol 3-kinase (PI 3-kinase) activity acts to inhibit calcium-dependent chloride secretion in T84 colonic epithelial cells. To further distinguish between the contributions of these two signaling pathways to the inhibition of secretion, we studied effects of insulin, because the insulin receptor links to PI 3-kinase but not to pathways postulated to generate Ins(3,4,5,6)P(4). Chloride secretion across T84 cell monolayers was studied in Ussing chambers. Activation of PI 3-kinase was assessed by Western blotting. Basolateral, but not apical, addition of insulin inhibited carbachol- and thapsigargin-induced chloride secretion in a time- and concentration-dependent fashion. Insulin-like growth factor-I (IGF-I) had similar effects. Insulin had no effect on Ins(3,4,5,6)P(4) levels, and the inhibitory effects of insulin and IGF-I on chloride secretion were fully reversed by the PI 3-kinase inhibitors wortmannin and LY-294002. Western blot analysis showed that both insulin and IGF-I recruited the 85-kDa regulatory and 110-kDa catalytic subunits of PI 3-kinase to anti-phosphotyrosine immunoprecipitates. In conclusion, insulin and IGF-I act to inhibit calcium-dependent chloride secretion through a PI 3-kinase-dependent pathway. Because insulin is released in a pulsatile fashion postprandially and IGF-I levels are elevated in pathological settings, our findings may have physiological and/or pathophysiological significance.

Androstadienes↗

PubMed2000

Dietary protein content affects the profiles of extracellular amino acids in the medial preoptic area of freely moving rats.

This study examined the effect of protein consumption on extracellular amino acid concentrations in the medial preoptic area (MPOA) of rats. Rats were given free access to diets containing 0, 25 or 50 % protein for 3-h duration, starting from the onset of dark cycle (1800 h). The microdialysis probe was implanted into the MPOA at 1500 h. Dialysates were collected every 20 min from 1700 h to 2100 h. Amino acid concentrations in dialysate samples were determined by reverse phase-HPLC. Extracellular amino acid concentrations in the MPOA were elevated by protein consumption within 20 to 40 min following the start of the meal. The 50 % protein diet resulted in increased (p<0.05) alanine, glutamine, isoleucine, leucine, methionine, tyrosine and valine concentrations, when compared with both baseline and the 0% protein diet. When the 25 % protein diet was fed, amino acid concentrations in the MPOA were between those after the 0 and 50% protein diets. The ratio of tryptophan to the total branched-chain amino acids in extracellular fluid was highest after the 0% protein diet and increased with time. We conclude that extracellular amino acid profiles in the MPOA are affected by dietary protein content.

Amino Acids↗

PubMed2000

Production of lactic acid by Lactobacillus rhamnosus with vitamin-supplemented soybean hydrolysate.

Batch fermentation studies were performed to evaluate the potentials of a complex nitrogen source, soybean, as an alternative to yeast extract for the economical production of lactic acid by Lactobacillus rhamnosus. An enzyme-hydrolysate of soybean meal, Soytone, with an adequate supplementation of vitamins was found to be highly effective in supporting lactic acid production from glucose and lactose. The effects of seven selected vitamins: d-biotin, pyridoxine, p-aminobenzoic acid, nicotinic acid, thiamine, pantothenic acid, and riboflavin, on cell growth and lactic acid production were investigated to provide the basis for the optimization of vitamin supplementation to minimize the cost. Pantothenic acid was the most required compound while the other six vitamins were also essential for high lactic acid productivity. As a result of the optimization, 15 g/l yeast extract could be successfully replaced with 19.3 g/l Soytone supplemented with the vitamins, resulting in a production of 125 g/l lactic acid from 150 g/l glucose. The volumetric productivity and lactate yield were 2.27 g/l/h and 92%, respectively, which were higher than those with 15 g/l yeast extract. The raw material cost was estimated to be 21.4 cent/kg lactic acid, which was only approximately 41% of that with yeast extract.

Journal Article↗

PubMed2000

Corneal insult affects the production and distribution of FGF-2 within the lacrimal gland.

The purpose of this study was to determine the distribution of FGF-2 within rabbit lacrimal glands and to determine whether corneal insult affects that distribution. The scarified corneas of experimental animals were inoculated either with adenovirus type 5 or buffer. Control animals were either untreated, or animals whose corneas were scarified. Twenty-one days later all animals were killed and the lacrimal glands were studied by immunocytochemistry and Western blotting to detect FGF-2. In untreated control animals, FGF-2 was immunolocalized predominantly within a population of elongated cells in the basal epithelium of ducts, and to a lesser degree in the basal epithelium of the acini. The elongated immunopositive cells appear to be myoepithelial cells known to be present at these sites. Interstitial cells around ducts and acini, and the basement membranes of the ducts and acini, were also immunopositive for FGF-2. Twenty-one days after adenovirus inoculation and scarification of the cornea, immunopositivity for FGF-2 was dramatically decreased in basement membranes, but increased within myoepithelial cells of the duct epithelium. These myoepithelial cells were frequently enlarged, bulging toward the duct lumen. In animals whose corneas were inoculated with buffer and scarified, or animals whose corneas were simply scarified, the changes in the lacrimal gland were similar, but somewhat less pronounced, to those of adenovirus-inoculated animals. Western blots confirmed the presence of FGF-2 immunoreactivity in all groups. The major band in untreated controls was at 24 kD, whereas all animals with corneal scarification had major bands at 38 kD. Densitometry of Western blots demonstrated that the amount of 24 kD FGF-2 present within the lacrimal gland after corneal scarification was at least 50% less than in untreated controls, whereas 38 kD FGF-2 was at least ten-fold greater. Our findings indicate that corneal scarification results in an altered distribution of FGF-2 within the lacrimal gland, which involves a decrease in low molecular weight FGF-2 and a dramatic increase in a higher molecular weight isoform of FGF-2. FGF-2 may be released from myoepithelial cells apically (exocrine) into the tear fluid and basally (autocrine/paracrine) into the connective tissue, as well as from extracellular complexes within basal laminae.

Adenoviridae Infections↗

PubMed2000

A comparison of 12- and 15-lead ECGS in ED chest pain patients: impact on diagnosis, therapy, and disposition.

The objective of this study was to investigate the diagnostic and therapeutic impact of the 15-lead electrocardiogram (15ECG) on the emergency department (ED) management of chest pain (CP) patients. The design was prospective use of 15ECG with real-time physician survey and retrospective comparison to 12-lead ECG (12ECG). The study took place in a University hospital ED. Adult CP patients participated. During the 15ECG period (June 1996 to July 1996), 595 patients (92% of CP patients) had 15ECG analysis. Diagnoses of acute coronary ischemic syndromes (ACIS) were as follows: 13 acute myocardial infarction (AMI, 7 anterior [ANT], 5 inferior [INF], 1 lateral [LAT], 2 posterior [POST], 1 right ventricular [RV]) and 136 unstable angina (USA) with 47% exhibiting ECG abnormality; the 2 POST and 1 RV AMI occurred in the setting of coexisting INF AMI. The following management strategies were used: 6 fibrinolytic therapy (TT), 4 primary angioplasty (PTCA), 67 rule-out myocardial infarction (ROMI), and 144 admission to critical care unit (CCU). During the 12ECG period (June 1995 to July 1995), 599 patients were encountered. The diagnoses of ACIS were as follows: 11 AMI (5 ANT, 4 INF, 2 LAT) and 146 USA with 51% exhibiting ECG abnormality (P = NS for diagnostic comparisons to 15ECG). The following management strategies were used: 5 TT, 5 PTCA, 59 ROMI, and 137 admission to CCU (P = NS for all treatment comparisons to 15ECG). Of 15ECG cases 81% had completed real-time physician survey, showing that the diagnosis and management ACIS were not altered by the 15ECG; physicians felt, however, that the 15ECG provided a more complete anatomic picture of the ACIS. No false-positive cases of additional lead STE were noted in this investigation except in cases involving abnormal intraventricular conduction such as the bundle branch block scenario. The 15ECG provided a more complete description of myocardial injury without altering the ED diagnosis, ED-based therapy, or hospital disposition in adult CP patients. Further study is required to identify patient subset(s) which may benefit from the 15ECG.

Angina, Unstable↗

PubMed2000

mu-conotoxin GIIIA interactions with the voltage-gated Na(+) channel predict a clockwise arrangement of the domains.

Voltage-gated Na(+) channels underlie the electrical activity of most excitable cells, and these channels are the targets of many antiarrhythmic, anticonvulsant, and local anesthetic drugs. The channel pore is formed by a single polypeptide chain, containing four different, but homologous domains that are thought to arrange themselves circumferentially to form the ion permeation pathway. Although several structural models have been proposed, there has been no agreement concerning whether the four domains are arranged in a clockwise or a counterclockwise pattern around the pore, which is a fundamental question about the tertiary structure of the channel. We have probed the local architecture of the rat adult skeletal muscle Na(+) channel (mu1) outer vestibule and selectivity filter using mu-conotoxin GIIIA (mu-CTX), a neurotoxin of known structure that binds in this region. Interactions between the pore-forming loops from three different domains and four toxin residues were distinguished by mutant cycle analysis. Three of these residues, Gln-14, Hydroxyproline-17 (Hyp-17), and Lys-16 are arranged approximately at right angles to each other in a plane above the critical Arg-13 that binds directly in the ion permeation pathway. Interaction points were identified between Hyp-17 and channel residue Met-1240 of domain III and between Lys-16 and Glu-403 of domain I and Asp-1532 of domain IV. These interactions were estimated to contribute -1.0+/-0.1, -0.9+/-0.3, and -1.4+/-0.1 kcal/mol of coupling energy to the native toxin-channel complex, respectively. mu-CTX residues Gln-14 and Arg-1, both on the same side of the toxin molecule, interacted with Thr-759 of domain II. Three analytical approaches to the pattern of interactions predict that the channel domains most probably are arranged in a clockwise configuration around the pore as viewed from the extracellular surface.

Amino Acids↗

PubMed1999

Cross-Canada spread of methicillin-resistant Staphylococcus aureus via transplant organs.

We report our investigation of the transmission of methicillin-resistant Staphylococcus aureus (MRSA) through transplantation. The kidneys, liver, and corneas were harvested from a child who died in Nova Scotia. Several days postmortem it was learned that culture of a premortem endotracheal tube aspirate from the donor yielded MRSA. Both kidneys were transplanted into a child in Nova Scotia and the liver into a child in Alberta. Both recipients subsequently became blood culture-positive for MRSA. One corneal ring from the donor was MRSA-positive. All four MRSA isolates were mecA-positive by polymerase chain reaction (PCR). The relatedness of the MRSA isolates was examined by restriction fragment length polymorphism (RFLP) analysis, a 16S-23S ribosomal PCR typing method, and comparison of antibiograms. Results were identical for all four MRSA isolates. These findings indicate that MRSA from the donor was transferred to recipients during implantation of harvested organs in Alberta and Nova Scotia, a cross-Canada spread.

Adolescent↗

PubMed1999

An intragastric amino acid mixture influences extracellular amino acid profiles in the lateral hypothalamic area of freely moving rats.

We tested the effect of equicaloric loads of glucose (0.89 g) or a balanced amino acid mixture (0.85 g) on extracellular amino acid concentrations in the brains of freely moving rats. At 15:30 hours, the microdialysis probe was inserted into the lateral hypothalamic area of ambulatory rats, and food and water were removed. Dialysates were collected every 20 min from 1 h prior to gavage (18:00 hours) and until 3 h after the gavage. Amino acid concentrations in the dialysate were determined by reverse-phase HPLC. Following the amino acid gavage, extracellular amino acid concentrations significantly increased from baseline for alanine, isoleucine, leucine, methionine, threonine, tyrosine, and valine. Those elevations occurred within 20-40 min following the amino acid load, and lasted up to 100 min. After the glucose and water treatments, amino acid concentrations were either not affected or gradually diminished from baseline. We conclude that extracellular amino acid concentration in the lateral hypothalamus is influenced by the composition of food consumed.

Administration, Oral↗

PubMed1998

A standardized protocol for the rapid preparation of bacterial DNA for pulsed-field gel electrophoresis.

A rapid method for the preparation of bacterial DNA for pulsed-field gel electrophoresis was developed for Gram-positive and Gram-negative bacteria. This method was accomplished by reducing the time for the cell lysis reaction, restriction endonuclease digestion, and electrophoresis to 1, 1.5, and 18 h, respectively. The whole procedure from the initial bacterial culture plate to the final analysis of restriction fragments can be completed within 24 h. This rapid method was successfully achieved for Staphylococcus aureus, Enterococcus faecalis, Neisseria gonorrhoeae, Salmonella typhimurium, Serratia marcescens, and Stenotrophomonas maltophilia.

Bacteriological Techniques↗

PubMed1996

Expanded genomic map of Campylobacter jejuni UA580 and localization of 23S ribosomal rRNA genes by I-CeuI restriction endonuclease digestion.

The genomic map of Campylobacter jejuni UA580 was expanded and more precisely constructed using I-CeuI, Sal/I and SmaI restriction endonucleases in conjunction with pulsed-field gel electrophoresis (PFGE). The presence of three fragments after digestion with I-CeuI confirmed the presence of three copies of the 23S ribosomal rRNA (rrl) gene. The genome size of Campylobacter jejuni UA580 was determined to be 1725 +/- 5.9 kbp by I-CeuI with fragment sizes of 1053 +/- 4.4, 361 +/- 2.7 and 311 +/- 3.6 kbp. Analysis of a PCR product from C. jejuni UA580 23S rRNA gene showed that I-CeuI did cut within the gene. The precise locations of the three genes were determined using I-CeuI with two copies of the 23S and 5S rRNA genes located separately from the 16S rRNA gene whereas the third copy of the 23S and 5S rRNA genes had a closer linkage to a 16S rRNA gene copy. Homologous gene probes were used to map additional genes and allowed the realignment of a few previously mapped genes on the chromosome. Other strains of C. jejuni were also cut into three fragments with I-CeuI, which generated variable PFGE patterns.

Campylobacter jejuni↗

PubMed1995

Genome map of Campylobacter fetus subsp. fetus ATCC 27374.

A physical map of the chromosome of Campylobacter fetus subsp. fetus was constructed by using pulsed-field gel electrophoresis of restriction fragments generated by SalI, SmaI and NotI. Digestion of the type strain ATCC 27374 with these restriction endonucleases resulted in generating 4-14 fragments. The order of the fragments was deduced from hybridization of these restriction fragments to Southern blots of pulsed-field gel electrophoresis gels generated by the other two enzymes. The estimated genome size was 1160 kb. The position of several homologous and heterologous genes was determined on the circular map. These included the 2.8-kb sapA gene, encoding the 97-kDa surface array protein. Three copies of ribosomal RNA genes for which the 16S, 23S and 5S rRNA appeared to be located in close proximity in each of the three regions. The RNA polymerase genes rpoA, rpoB, and rpoD were mapped and appeared to be situated close together in one region. The flagellin genes (flaAB) of C. jejuni and the gyrase genes gyrA and gyrB of C. perfringens and Bacillus subtilis, respectively, were used to identify the locations of flaAB, the gyrA and the gyrB genes on the ATCC 27374 chromosome.

Base Sequence↗

PubMed1995

Microdialysis as a tool to measure dietary and regional effects on the complete profile of extracellular amino acids in the hypothalamus of rats.

Regional and dietary-induced changes in hypothalamic extracellular amino acid concentrations were examined. Microdialysis probes were simultaneously implanted in the hypothalamic paraventricular nucleus (PVN) and the lateral hypothalamus (LH) of anesthetized rats and perfused at a rate of 2 microliters/min. Dialysates were collected every 20 min for 1 h prior to gavage of a balanced amino acid mixture (0.85 g patterned after 1 g of chicken egg albumin) and then every 20 min for 3 h after treatment. Tail vein blood samples were also collected. Marked changes in plasma levels of most amino acids were evident immediately following the amino acid gavage. In the PVN, concentrations of isoleucine, leucine, methionine and valine all increased within 40 min, whereas significant decreases in glutamine, histidine and taurine were observed in the LH. In a separate study, PVN extracellular amino acid concentrations were examined in awake, freely-behaving rats following gavage of equicaloric loads of a balanced amino acid mixture, glucose (0.89 g) or water. Dialysate levels of glutamate, isoleucine, leucine, methionine, threonine, tyrosine and valine showed reliable increases after amino acid treatment, although the overall time course of these effects differed somewhat. The amino acid profile of the PVN was, in general, unaffected by glucose administration. These findings suggest that specific brain regions may respond uniquely to amino acid ingestion and further imply that dietary composition may influence the amino acid profiles of the extracellular fluid in brain.

Amino Acids↗

PubMed1995

Characterization of chromosomal DNA profiles from Helicobacter pylori strains isolated from sequential gastric biopsy specimens.

The restriction endonuclease profiles of DNAs from Helicobacter pylori strains isolated from 20 patients in two or more consecutive biopsy specimens over a period of up to 2 years were analyzed by pulsed-field gel electrophoresis with NotI and NruI. H. pylori strains possess a high degree of genomic diversity which was not observed to occur in vivo, and attempts to observe it in vitro were not successful.

DNA, Bacterial↗

PubMed1994

Genome conservation in Helicobacter mustelae as determined by pulsed-field gel electrophoresis.

Genomic DNA from 15 strains of Helicobacter mustelae was subjected to pulsed-field gel electrophoresis (PFGE) after digestion with PacI and SfiI. H. mustelae genome DNA appeared very similar in all strains examined, whether isolated from ferrets or mink or from animals bred in either the USA or in the UK. The H. mustelae genome size was estimated to be 1.7 Mb, similar in size to that of H. pylori. A minor difference in PacI PFGE pattern and genome size was observed between rifampicin-resistant and rifampicin-susceptible derivatives of H. mustelae F251. Another minor difference in genome pattern based on PFGE with SfiI was observed between an H. mustelae strain used to experimentally infect four ferrets which resulted in loss of an SfiI site in strains obtained from the newly infected ferrets. Thus, although minor differences in PFGE pattern were noted, H. mustelae lacks the genomic diversity observed in H. pylori.

Animals↗

PubMed1992

Construction of a Helicobacter pylori genome map and demonstration of diversity at the genome level.

Genomic DNA from 30 strains of Helicobacter pylori was subjected to pulsed-field gel electrophoresis (PFGE) after digestion with NotI and NruI. The genome sizes of the strains ranged from 1.6 to 1.73 Mb, with an average size of 1.67 Mb. By using NotI and NruI, a circular map of H. pylori UA802 (1.7 Mb) which contained three copies of 16S and 23S rRNA genes was constructed. An unusual feature of the H. pylori genome was the separate location of at least two copies of 16S and 23S rRNA genes. Almost all strains had different PFGE patterns after NotI and NruI digestion, suggesting that the H. pylori genome possesses a considerable degree of genetic variability. However, three strains from different sites (the fundus, antrum, and body of the stomach) within the same patient gave identical PFGE patterns. The genomic pattern of individual isolates remained constant during multiple subcultures in vitro. The reason for the genetic diversity observed among H. pylori strains remains to be explained.

Base Sequence↗

PubMed1992

Genome maps of Campylobacter jejuni and Campylobacter coli.

Little information concerning the genome of either Campylobacter jejuni or Campylobacter coli is available. Therefore, we constructed genomic maps of C. jejuni UA580 and C. coli UA417 by using pulsed-field gel electrophoresis. The genome sizes of C. jejuni and C. coli strains are approximately 1.7 Mb, as determined by SalI and SmaI digestion (N. Chang and D. E. Taylor, J. Bacteriol. 172:5211-5217, 1990). The genomes of both species are represented by single circular DNA molecules, and maps were constructed by partial restriction digestion and hybridization of DNA fragments extracted from low-melting-point agarose gels. Homologous DNA probes, encoding the flaAB and 16S rRNA genes, as well as heterologous DNA probes from Escherichia coli, Bacillus subtilis, and Haemophilus influenzae, were used to identify the locations of particular genes. C. jejuni and C. coli contain three copies of the 16S and 23S rRNA genes. However, they are not located together within an operon but show a distinct split in at least two of their three copies. The positions of various housekeeping genes in both C. jejuni UA580 and C. coli UA417 have been determined, and there appears to be some conservation of gene arrangement between the two species.

Bacterial Proteins↗