Biomedical subjects

Y M Kwon

Publications and source records attributed to Y M Kwon.

At least 19 recordsLinked to original sources

PubMed2005

Reduction of Salmonella enterica serovar enteritidis colonization and invasion by an alfalfa diet during molt in Leghorn hens.

The standard method for molting to stimulate multiple egg-laying cycles in laying hens is feed deprivation. However, the physiological changes within hens caused by feed deprivation increase susceptibility of the hens to Salmonella enterica serovar Enteritidis (SE) infection. In an effort to develop an alternative method to induce molting without increasing susceptibility to SE, an alfalfa diet was compared with the standard molting method for the level of ovary regression and SE colonization. Hens over 50 wk of age were divided into 3 treatment groups (12 hens/group): nonmolting by normal feeding (NM), molting by feed deprivation (M), and molting by alfalfa diet (A). Individual hens on all treatments were challenged orally with 10(5) cfu of SE on the fourth day after feed changes and were analyzed for ovary weight and SE colonization or invasion in crop contents, cecal contents, liver, spleen, and ovary on the ninth day. In 3 of the 4 trials, there was a significant decrease in SE colonization of the crop between the alfalfa diet (A) and the feed deprived molt (M). In most of the 4 trials, there was a significant reduction in SE infected organs in birds fed the alfalfa diet (A) compared with birds undergoing feed deprived molt (M). Most of the trials showed no significant difference in overall SE between A and NM. Therefore, the results of this study suggest that an alfalfa diet has the potential to be used as an alternative method for forced molting, without increasing the incidence of SE in eggs and internal organs.

Animal Feed↗

PubMed2003

Use of a Salmonella typhimurium hilA fusion strain to assess effects of environmental fresh water sources on virulence gene expression.

Many fruits and vegetables are irrigated with water from rivers, lakes and even wastewater systems. Irrigation may be a route for the introduction of Salmonella. Our objectives in this study were to determine survivability and virulence expression in a strain of Salmonella typhimurium when exposed to environmental water sources. Virulence expression was measured using a beta-galactosidase assay on a hilA:lacZY fusion strain of S. typhimurium. Water samples for environmental impact studies were taken from a local pond and specific sites along the Rio Grande River, which serves as a source of irrigation water in southern Texas. There was a significant difference (p<0.05) of virulence expression among the water sites. Certain regions along the Rio Grande River yielded greater amounts of beta-galactosidase activity than others. All sites yielded at least a two-fold greater virulence response than S. typhimurium grown in brain heart infusion. Salmonella survivors were enumerated as colony forming units (CFU)/ml as plated on a selective medium for the duration of 1 week and beta-galactosidase assays were performed to determine a possible correlation between culturable cells and virulence gene expression. Bacterial cells remained viable but decreased after 7 days incubation. In conclusion, water sampled at specific locations and at different times water samples exhibited differences in virulence expression in S. typhimurium.

Agriculture↗

PubMed2003

Isolation of Salmonella typhimurium Tn5 mutants defective for survival on egg shell surface using transposon footprinting.

The objective of this study was to screen the genome of Salmonella typhimurium for genes potentially required for survival on egg shell surfaces using a novel transposon footprinting method based on a previously developed Tn5 mutagenesis technique. Since the amplified DNA fragment length should usually be unique for each mutant, the polymerase chain reaction products separated on an agarose gel generate a transposon footprint with each band in the footprint representing the corresponding Tn5 mutant. By comparing the footprints from the pools of S. typhimurium Tn5 mutants before and after inoculation on egg shell surfaces, Tn5 mutants not recovered after selection were rapidly identified for potential isolation and genetic analysis.

Animals↗

PubMed2001

cDNA cloning of two isoforms of ornithine carbamoyltransferase from Canavalia lineata leaves and the effect of site-directed mutagenesis of the carbamoyl phosphate binding site.

The immunoscreening method was used to isolate cDNAs of 1323 bp (ClOCT1) and 1433 bp (ClOCT2) encoding two ornithine carbamoyltransferases (OCT, EC 2.1.3.3) from the cDNA expression library of Canavalia lineata leaves constructed in a lambdaZAP Express vector. ClOCT1 and ClOCT2 encode 359 and 369 amino acids, respectively. The N-terminals of deduced amino acid sequences of the two cDNAs showed typical features of the transit peptide of chloroplast targeting proteins. The ornithine-binding domain (FMHCLP) and catalytic domain (HPXQ) of ClOCT1 and ClOCT2 and the carbamoyl phosphate (CP)-binding site of ClOCT1 (SMRTR) are identical to OCTs of other plant species, pea and Arabidopsis thaliana. However, the CP-binding site sequence of ClOCT2, SLRTH, has not yet been reported. Both ClOCT1 and ClOCT2 cDNAs were expressed in Escherichia coli BL21 (DE3) by using expression vector pET30a. Recombinant ClOCT1 protein showed 14 times higher ornithine-dependent OCT activity than canaline-dependent OCT activity. In contrast, recombinant ClOCT2 protein showed 13 times higher canaline-dependent OCT activity than ornithine-dependent OCT activity. The two amino acids of the CP-binding site of ClOCT2 (SLRTH) were combinatorially changed to those of the CP-binding site of ClOCT1 (SMRTR) by site-directed mutagenesis. When Leu-118 of ClOCT2 was changed to Met, ornithine-dependent activity was increased significantly. It is assumed that the substrate specificity of ClOCT1 or ClOCT2 proteins partially depends on the amino acid sequence of the CP-binding site.

Amino Acid Sequence↗

PubMed2001

In situ study of insulin aggregation induced by water-organic solvent interface.

PURPOSE: The aim of this study was to assess insulin stability by monitoring in situ time-course of insulin aggregation induced by a water-organic solvent (o/w) interface that occurs during the microencapsulation process. METHODS: Insulin aggregation at a simple o/w interface was monitored spectrophotometrically by detecting the percentage of turbidity changes (%T) at 350 nm. The effects of protein concentration and agitation and the presence of poly (lactic-co-glycolic acid) (PLGA) in methylene chloride (MC) on insulin aggregation were observed. For the 0.72 mg/ml insulin in phosphate-buffered saline (PBS), the effect of nonionic (dodecyl maltoside [DDM]) and anionic (sodium dodecyl sulfate [SDS]) surfactant in PBS were also evaluated at various protein/surfactant mol ratios. The conformation of insulin protected by a 10-fold molar excess of SDS recovered after 1 h of contact with MC was evaluated via circular dichroism (CD) spectroscopy. RESULTS: A typical turbidity-time profile was represented by a sigmoidal curve. Greater change in %T was observed with increasing insulin concentration, in the presence of PLGA in MC and in the presence of agitation. DDM failed to delay insulin aggregation at all ratios used, whereas a less than 10% change in %T was observed in 1 h when a 10- - approximately 20-fold excess of SDS was used. CD spectra indicated that the presence of insulin in SDS after 1 h of contact with MC qualitatively retained its secondary structure integrity. CONCLUSIONS: An experimental method was designed for an in situ assessment of protein stability at the o/w interface.

Chemical Phenomena↗

PubMed2001

Scintigraphic assessment of the acetabulum after arthroplasty with reference to cup geometry.

A unique scintigraphic appearance has been noted in uncomplicated hip prostheses which have the acetabular component inserted after under-reaming or where a cone-shaped cup is inserted. We hypothesized that the pattern of increased uptake around the superior and inferior third of the cup with relatively reduced uptake in the medial third is due to preferential loading of the rim of the cup. A prospective study of ten patients with uncomplicated hip prostheses supports the hypothesis.

Acetabulum↗

PubMed2001

Characterization of prejunctional purinoceptors inhibiting noradrenaline release in rat mesenteric arteries.

The effects of purinoceptor agonists on noradrenaline NA release by electrical stimulation in rat mesenteric arteries were examined to clarify the pharmacological properties of prejunctional purinoceptors on adrenergic nerves. Adenosine and the other P1-receptor agonists, 5'-(N-ethylcarboxamido) adenosine and 2-chloroadenosine, significantly inhibited the release of NA. Also beta,gamma-methylene ATP and 2-methylthio ATP, P2-receptor agonists, significantly inhibited NA releases. The inhibitory effect of adenosine was significantly reduced by adenosine deaminase, but those of beta,gamma-methylene ATP and 2-methylthio ATP were not affected. This suggests that the inhibitory effects of P2-receptor agonists are not due to conversion into adenosine. 1,3-Dipropyl-8-cyclopentylxanthine (DPCPX), a P1 (A1)-receptor antagonist, significantly reduced the inhibitory effects of not only the P1- but also P2-receptor agonists. Therefore, DPCPX appears to act on both prejunctional P1- and P2-receptor as an antagonist. Pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), a P2-receptor antagonist, significantly reduced the inhibitory effects of the P2-receptor agonists, but not those of the P1-receptor agonists. From these findings in the rat mesenteric artery, the P1-receptor agonist-induced inhibition of NA-release appears to be mediated via a well-known prejunctional P1-receptor of the A1-subtype, but the P2-receptor agonist-induced inhibition appears to be mediated via an unidentified purinoceptor that is blocked not only by P2-receptor antagonists but also by P1-receptor antagonists.

2-Chloroadenosine↗

PubMed2001

Anastomosis of vessels less than 2 mm with the vascular clip system clip applier.

Sutures may cause endothelial trauma and occlusion. The vascular clip system (VCS) clip applier may minimize endothelial injury. Fourteen carotid arteries of nine adult rabbits were transected and re-anastomosed with either #7-0 polypropylene (Group I, n=8) or VCS clips (Group II, n=6). The animals were sacrificed at 1, 3, 8, 14, and 30 days postoperatively. The operation time and bleeding amount were checked for each anastomosis. Carotid angiograms, photography, H&E staining and scanning electron microscopy (SEM) were performed. Fibrin and thrombus, inflammatory cell infiltration, endothelial disruption, luminal distortion, fibrosis, and wall thickening were compared. The luminal diameter was greater in group II. There were minimal differences in thrombosis, wall thickening and fibrosis between the two groups. However, fibrin, inflammatory cell infiltration, multinucleated giant cell formation, endothelial disruption, and luminal distortion were greater in group I. On SEM, group I showed trans-mural penetration. In contrast, group II showed suture margin eversion and no transmural penetration. Stenosis was greater in group I than in group II on carotid angiogram. The operation time was shorter in group II than in group I, i.e. 5+/-1.4 min vs. 11+/-3.8 min, respectively. The current data showed similar or superior results with VCS clips in comparison to conventional suturing with polypropylene.

Angiography↗

PubMed2000

Two closely related cDNAs encoding actin-depolymerizing factors of petunia are mainly expressed in vegetative tissues.

Actin-depolymerizing factor (ADF) is one of the small actin-binding proteins that regulate actin dynamics in cells. We have isolated two cDNA clones, PhADF1 and PhADF2, encoding ADF from cDNA libraries constructed from petal protoplast cultures and flowers of Petunia hybrida. PhADF1 and PhADF2 encode polypeptides of 139 and 143 amino acids with a calculated molecular mass of 16.04 and 16.51kDa, respectively. Co-sedimentation assay showed that the recombinant PhADF1 protein produced in Escherichia coli binds to F-actin at pH7. 0 and preferentially depolymerizes it at pH8.0. Gene tree analysis indicates that the plant ADF family can be grouped into four classes, and PhADFs are included in class I. Southern blot analyses revealed that one or two copies of PhADF genes are present in petunia genome, and several other related isoforms also exist. Northern blot analyses indicated that PhADF1 and PhADF2 are closely related and abundantly expressed in every plant organ except pollen. In addition, they are highly accumulated in mature vegetative tissue (petal, leaf, and stem). Our results indicate that the transcription of petunia ADF genes is differentially regulated by developmental signals.

Actin Depolymerizing Factors↗

PubMed2000

The AGAMOUS-LIKE 20 MADS domain protein integrates floral inductive pathways in Arabidopsis.

The very late-flowering behavior of Arabidopsis winter-annual ecotypes is conferred mainly by two genes, FRIGIDA (FRI) and FLOWERING LOCUS C (FLC). A MADS-domain gene, AGAMOUS-LIKE 20 (AGL20), was identified as a dominant FRI suppressor in activation tagging mutagenesis. Overexpression of AGL20 suppresses not only the late flowering of plants that have functional FRI and FLC alleles but also the delayed phase transitions during the vegetative stages of plant development. Interestingly, AGL20 expression is positively regulated not only by the redundant vernalization and autonomous pathways of flowering but also by the photoperiod pathway. Our results indicate that AGL20 is an important integrator of three pathways controlling flowering in Arabidopsis.

AGAMOUS Protein, Arabidopsis↗

PubMed2000

Genomic structure of ornithine carbamoyltransferase gene from Canavalia lineata.

The nucleotide sequences of the complete ornithine carbamoyltransferase (OCT) gene containing 3,964 bp 3' region and 1,701 bp promoter region were determined from Canavalia lineata leaves. The exons range in size from 131 bp to 390 bp, while the introns range in size from 102 bp to the relatively large size of 1,465 bp; all the splicing junctions followed the conserved GT/AG rule. The transcription initiation site is located 12 bp upstream of the ATG translation initiation site and 69 bp downstream of TATA-box. The regulatory elements located in the 1,701 bp 5' upstream region are GT-1, GATA motif, I-box, SBF-1, Nit2 etc. The structure of the OCT gene from C. lineata was compared with Arabidopsis thaliana, in which two genes consist of 5 exons and 4 introns. The nucleotide sequence of exons and deduced amino acid sequence exhibits 70% and 69% homology, respectively. The deduced amino acid sequences show identical conserved "HPXQ" box, carbamoyl phosphate and ornithine binding sites. The introns of C. lineata OCT gene are larger than those of A. thaliana, except for intron 3. Genomic DNA blot analysis showed that a single ornithine-dependent OCT gene exists in the C. lineata genome.

Amino Acid Sequence↗

PubMed2000

A bioluminescent Escherichia coli auxotroph for use in an in vitro lysine availability assay.

Microbiological methods have been used to determine the amino acid availability of a variety of animal feed and human food protein sources. Growth of Escherichia coli auxotrophs have been shown to yield a consistent linear response to lysine concentration when compared to chemical measures. Extent of total growth of E. coli lysine mutant (American Type Culture Collection #23812) when measured as optical density (OD) displays a lysine-dependent growth response that can be used to estimate lysine in feed proteins. However, typical OD-based growth studies for amino acid quantitation using the mutant may require anywhere from 12 to over 40 h. To develop an improved rapid method for lysine quantitation in protein sources, the plasmid pJHD500 carrying genes that encode for expression of bioluminescence and ampicillin resistance was transformed into the E. coli mutant by electroporation (set at 1.80 kV). The luminescence measured during early exponential growth allowed detectable differentiation of lysine concentration in the media in 4 h. When the luminescence method was compared with the conventional optical density lysine growth assay, the correlation coefficient was 0.989. Lysine availability valued for enzymatically hydrolyzed protein sources were comparable with availability measures using animal methods for lysine availability. This research shows potential applications for more rapid quantitative measurement of bioavailable lysine.

Animal Feed↗

PubMed2000

Efficient amplification of multiple transposon-flanking sequences.

Transposon mutagenesis is a very useful tool for gene identification in bacteria. Once the transposon mutants of interest are isolated, it is often necessary to identify the sequences that flank the transposon insertions. We devised an efficient method for specific amplification of transposon-flanking sequences that requires the sequence information of only transposon-specific sequences. The basic steps for this method consists of (1) digestion with a restriction enzyme, (2) ligation with a Y-shaped linker and (3) polymerase chain reaction amplification using a transposon-specific primer and a primer specific to the Y-shaped linker. The feasibility of this method was demonstrated with mini-Tn5 mutants of Salmonella typhimurium. We also found that this method can be used for simultaneous amplification of multiple transposon-flanking sequences.

3' Untranslated Regions↗

PubMed2000

A new technique of subtrochanteric shortening in total hip arthroplasty: surgical technique and results of 9 cases.

Total hip arthroplasty for severe chronic proximal femoral migration, most commonly seen in congenital dislocation of the hip, has been associated with high rates of complications. A new technique of femoral subtrochanteric shortening osteotomy with the prosthesis in situ is described. This technique minimizes the potential complications, allows for correction of severe femoral neck anteversion, and gives excellent rotational stability, while preserving the proximal femur for better press-fit cementless fixation. In this series, there were 9 cases: 6 women and 2 men with a mean age of 53 years (range, 26-77 years). The average follow-up period was 56 months (range, 6-86 months). The mean preoperative Harris Hip Score was 31 (range, 20-35), and the mean postoperative score was 81 (range, 60-98). At follow-up, all patients reported significant pain relief and functional improvement. All osteotomies appeared to be healed on radiographs by 12 weeks. There were 3 complications. The first complication was a recurrent dislocation resulting from muscle incompetence, which was revised using a constrained liner and a 32-mm head with no further dislocations. The second complication was a breach of the femoral shaft, which was treated operatively using a longer stem. The third complication was a proximal femoral shaft split, which was treated by leaving the cerclage wire in situ. This technique should be considered in cases of congenital dislocation of the hip and when femoral shortening is needed.

Adult↗

PubMed2000

Recovery of a marker strain of Salmonella typhimurium in litter and aerosols from isolation rooms containing infected chickens.

Screening of poultry flocks for foodborne pathogen Salmonella contamination is critical for Salmonella control in preharvest stages of poultry production. In this study, two sampling methods (litter and air filter) were compared for detection of S. typhimurium from experimentally infected chicks some of which had received either a probiotic competitive exclusion culture or transfer of cecal contents from salmonellae-free adult birds. At 4, 9, and 11 days after inoculation, S. typhimurium samples were enumerated by selective plating. For both types of sampling, the control birds yielded the greatest levels of environmental contamination followed by the samples from the probiotic inoculated birds with the birds receiving the cecal transfer culture having the lowest levels of contamination. Although the two sampling methods responded in a similar fashion, detection sensitivity needs to be increased for air filter sampling.

Aerosols↗

PubMed1999

Analysis of cDNAs expressed during first cell division of petunia petal protoplast cultures using expressed sequence tags.

A large-scale sequence analysis of randomly selected cDNA clones was performed to isolate numerous genes in petunia petal protoplast cultures. We have partially sequenced 1158 randomly selected genes of the cDNA library constructed from 2-6 d cultured petal protoplasts. Three hundred and sixty-five different genes were identified, 25% of which showed significant similarity to existing sequences in the petunia, and an array of other organisms. In this report, 90 independent genes are analyzed in detail. A functional categorization of the database-matched expressed sequence tags (ESTs) showed that defense- or stress-related genes, as well as genes involved in the primary metabolic pathways and in the transcriptional or translational apparatus are abundantly represented. In particular, ESTs were identified with apparent homologies to the cyclin-dependent kinase, histone, actin-depolymerizing factor, proteasome, and ubiquitin which are expected to be related to cell division or to cell cycle control.

Cell Division↗

PubMed1999

Primary CD56 + nasal-type T/natural killer-cell subcutaneous panniculitic lymphoma: presentation as haemophagocytic syndrome.

Natural killer (NK) cells are large granular lymphocytes that mediate cytotoxic reactions which are not restricted by the major histocompatibility complex. In recent years it has become apparent that a minor proportion of malignant lymphomas expresses an NK-cell phenotype defined by its reactivity with the CD56 antibody. Primary purely cutaneous CD56 + lymphomas have rarely been reported. They share a generally aggressive course and are highly associated with Epstein-Barr virus. We describe a patient with a primary cutaneous nasal-type T/NK-cell lymphoma that presented as a haemophagocytic syndrome and showed an aggressive clinical course.

Adult↗

PubMed1999

The hypotensive effect of docosahexaenoic acid is associated with the enhanced release of ATP from the caudal artery of aged rats.

Fish oils have been shown to lower blood pressure in hypertensive subjects. To determine the mechanism of this hypotensive effect, we examined the effects of docosahexaenoic acid (DHA), one of the (n-3) polyunsaturated fatty acids in fish oil, on blood pressure and on the release of adenyl purines, such as ATP, ADP, AMP and adenosine, from the caudal arteries of aged rats. Aged female Wistar rats (100 wk) were fed a high cholesterol diet and were administered intragastrically ethyl all-cis-4,7,10,13,16,19-docosahexaenoate [300 mg/(kg.d)] for 12 wk (DHA group) or vehicle alone (control group). Compared with the controls, rats supplemented with DHA had significantly greater (10.1%) DHA concentrations in the caudal arteries. This was associated with more total (n-3) arterial fatty acids, a greater unsaturation index of arterial fatty acids, 43.9% lower plasma noradrenaline levels and the repression of the elevation in blood pressure observed with advancing age. The amount of purines released, both spontaneously and in response to noradrenaline, from arterial segments of DHA-supplemented rats was significantly higher than that released from tissues of control rats. Regression analysis revealed significant negative relationships between the total amount of purines released from the artery and the systolic (SBP) and diastolic (DBP) blood pressures. These results suggest that in aged rats, supplementation with DHA alters the membrane fatty acid composition as well as the amount of ATP released from vascular endothelial cells and decreases plasma noradrenaline, and that these factors may ameliorate the rise in blood pressure normally associated with advancing age.

Adenosine Triphosphate↗