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Biomedical subjects

Y Kim

Publications and source records attributed to Y Kim.

At least 451 records · Page 25Linked to original sources

Features for a B-ISDN telemedicine system and its application.

Telemedicine technology is emerging as a new way of medical practice. It will provide more cooperative activity between departments and more comfortable access for disabled patients home. We developed two types of telemedicine system: a telediagnosis system and a home care system. Our telemedicine application is aimed to be run on the broadband-integrated services digital network (B-ISDN). The legacy network is also considered. The user interface is designed to help doctors to communicate easily. The key elements of telemedicine systems are user friendly interface, medical multimedia database design highly refined display technique.

Computer Communication Networks↗

Web based 3-D medical image visualization on the PC.

With the recent advance of Web and its associated technologies, information sharing on distribute computing environments has gained a great amount of attention from many researchers in many application areas, such as medicine, engineering, and business. One basic requirement of distributed medical consultation systems is that geographically dispersed, disparate participants are allowed to exchange information readily with each other. Such software also needs to be supported on a broad range of computer platforms to increase the softwares accessibility. In this paper, the development of world-wide-web based medical consultation system for radiology imaging is addressed to provide platform independence and greater accessibility. The system supports sharing of 3-dimensional objects. We use VRML (Virtual Reality Modeling Language), which is the defacto standard in 3-D modeling on the Web. 3-D objects are reconstructed from CT or MRI volume data using a VRML format, which can be viewed and manipulated easily in Web-browsers with a VRML plug-in. A Marching cubes method is used in the transformation of scanned volume data sets to polygonal surfaces of VRML. A decimation algorithm is adopted to reduce the number of meshes in the resulting VRML file. 3-D volume data are often very large in size, hence loading the data on PC level computers requires a significant reduction of the size of the data, while minimizing the loss of the original shape information. This is also important to decrease network delays. A prototype system has been implemented (http://cybernet5.snu.ac.kr/-cyber/mrivrml .html), and several sessions of experiments are carried out.

Algorithms↗

[Long-term survival in a case of multiple liver metastasis from postoperative gastric cancer effectively treated by hepatic intraarterial infusion chemotherapy using MMC and pirarubicin].

We experienced a case of multiple liver metastasis from postoperative gastric cancer who showed long-term survival with hepatic arterial infusion chemotherapy (HAI) of MMC and pirarubicin. A catheter was inserted into the hepatic artery, and 4 mg of MMC and 20 mg of pirarubicin were administered through an implantable port catheter every two to four weeks. The total dose of MMC and pirarubicin by the time of this report was 164 mg and 820 mg, respectively. The follow-up CT scan 2 months after the beginning of HAI showed a decrement of the liver tumors. The decrease rate at 12 and 17 months was 50% and 70%, respectively, which was diagnosed as partial response (PR). The therapeutic effect at 49 months is still PR without any sign of tumor enlargement of extra hepatic lesion.

Antineoplastic Combined Chemotherapy Protocols↗

Angiotensin I converting enzyme gene polymorphisms in systemic lupus erythematosus: decreased prevalence of DD genotype in African American patients.

The presence of the D (deletion) allele at the angiotensin converting enzyme (ACE) gene has been associated with a) adverse vascular events contributing to early mortality and b) progressive deterioration of renal function in a variety of chronic glomerular diseases. We investigated the potential role of ACE polymorphisms in patients with systemic lupus erythematosus (SLE). Two hundred and sixteen (216) SLE patients (121 Caucasians; 78 African Americans; and 17 other) and 200 normal controls were studied; 134 patients had evidence of renal disease. ACE genotypes were determined by a polymerase chain reaction based assay. The frequency of genotype DD was increased in African American normal controls compared to Caucasians (55% vs. 37%, p = 0.017) and in African American normal controls vs. African American lupus patients (55% vs. 30%, p = 0.008). Trend analysis of the genotype distribution across the three African American groups (renal, non-renal, controls) revealed a trend of increased frequency of I and decreased frequency of D as likelihood of renal disease increases (p = 0.008). No association between any ACE genotype with parameters of renal disease and/or response to therapy was identified. African American patients with lupus have a lower frequency of DD genotype as compared to African American normal controls. Further studies will be necessary to address whether this is due to decreased survival of these patients, a protective effect of DD genotype from developing the disease or a chance sample effect.

Adult↗

[A case of locally advanced breast cancer successfully treated with intra-arterial infusion chemotherapy of high-dose epirubicin].

A case of locally advanced breast cancer was treated with intra-arterial infusion chemotherapy using high-dose epirubicin. The 1st and 2nd cycle consisted of 210 mg (day 1, 4, 7) and 150 mg (day 1, 4, 7), respectively. After completion of 2 cycles of the regimen, remarkable loco-regional control and improved QOL were obtained. Leucopenia (nadir; 1,000/mm3) was the dose limiting factor, but well-tolerated with supportive therapy. The patient underwent salvage surgery and maintenance intra-venous chemotherapy (epirubicin; 30 mg/3 weeks). At this writing, the patient is enjoying favorable QOL. These findings suggested that this modality was effective for cases with exctensive loco-regional involvement.

Antibiotics, Antineoplastic↗

Nucleotide sequence of the Pseudomonas sp. DJ77 phnG gene encoding 2-hydroxymuconic semialdehyde dehydrogenase.

The nucleotide sequence of a 1520 bp region, spanning the coding region for the meta-cleavage pathway enzyme, 2-hydroxymuconic semialdehyde dehydrogenase, was determined. This enzyme, encoded by the phnG, is the first of three sequential enzymes required for conversion of 2-hydroxymuconic semialdehyde, which is produced from catechol by the PhnE catechol 2,3-dioxygenase, to 2-hydroxypent-2,4-dienoate in the dehydrogenative branch of the pathway. The deduced protein sequence is 484 amino acid residues long with a M(r) of 51504. The phnG has a high degree of homology with genes encoding isofunctional proteins from other Pseudomonas strains. We now show that the relative position of the phnG dehydrogenase gene in the phn operon is unique compared to the other meta-cleavage operons which have a dehydrogenative branch of the pathway.

Alcohol Oxidoreductases↗

Cloning and characterization of the regulatory genes phlR1 and phlR2 involved in phenol metabolism from Alcaligenes eutrophus JMP134.

One mutant (AEK201) of Alcaligenes eutrophus JMP134 deficient in phenol metabolism was isolated by transposon mutagenesis using pSUP2021, a suicide plasmid. The 14.5 kb EcoRI fragment containing Tn5 and flanking DNA was cloned from AEK201 and used to probe a gene bank of wild type by colony hybridization. All five positive cosmids isolated rendered AEK201 to grow on phenol. The data from subcloning revealed that a trans-acting factor encoded on the 2.3 kb SalI-HindIII fragment, which is common to all cosmids, allowed the mutant to restore three enzyme activities tested (phenol hydroxylase, catechol 1,2-dioxygenase, and catechol 2,3-dioxygenase). This fragment seems to act as a positive regulator on the entire phenol pathway. Another regulatory segment was subcloned from the 16.8 kb HindIII fragment on which phenol hydroxylase and catechol 2,3-dioxygenase activities were carried [Kim, Y., Ayoubi, P., and Harker, A. R. (1996) Appl. Environ. Microbiol. 62, 3227-3233]. The expression of phenol hydroxylase activity was entirely repressed in the presence of this segment in Pseudomonas aeruginosa PAO1c, but the enzyme activity was increased in A. eutrophus AEK301, suggesting that this trans-acting factor is both an activator and a repressor for phenol hydroxylase. Possible regulatory mechanisms for the phenol pathway in A. eutrophus JMP134 are discussed.

Alcaligenes↗

Protection of canine pancreatic microvascular endothelium against cold ischemic injury during preservation by the two-layer method.

BACKGROUND: Ischemic injury of the microvascular endothelium during cold preservation causes a disturbance of vascular microcirculation after reperfusion and results in graft failure. Recently we have shown that oxygenation of the canine pancreas during preservation by the two-layer method extends the period of preserved pancreatic viability. The aim of this study was to clarify the role of the oxygenation of the pancreas graft by the two-layer method in the viability of the microvascular endothelium during preservation. METHODS: After preservation of the canine pancreas by simple cold storage in Euro-Collins solution (EC) (group 1) or by the two-layer method using EC (group 2) for 48 hr, the viability of vascular endothelial cells was judged from nuclear trypan blue uptake. Pancreatic tissue perfusions were measured with a hydrogen gas clearance technique, and graft survival rates were examined after autotransplantation. In the control group, the grafts were autotransplanted without preservation (group 3). RESULTS: Graft survival rates were 0 of 5 (0%), 5 of 5 (100%), and 5 of 5 (100%) in groups 1, 2, and 3, respectively. The percentage of trypan blue-positive vascular endothelium in group 1 was significantly higher compared with group 3 (control) (26.4+/-1.7 vs. 7.4+/-4.3%, P<0.01). The two-layer method (group 2) decreased trypan blue uptake (11.3+/-3.7 vs. 26.4+/-1.7%, P<0.01). Pancreatic tissue perfusions after 2 hr of reperfusion were in inverse proportion to trypan blue uptake. Namely, pancreatic tissue perfusions in group 1 were significantly lower than group 3 (control) (45.6+/-12.8 vs. 64.5+/-20.6 ml/min/100 g, P<0.01). The two-layer method (group 2) improved pancreatic tissue perfusions (68.8+/-8.6 vs. 45.6+/-12.8 ml/min/100 g, P<0.01). CONCLUSION: We conclude that oxygenation of the pancreas during preservation by the two-layer method protects the microvascular endothelium from cold ischemic injury. Consequently, pancreatic microcirculation can be maintained after reperfusion, thus extending the period of preserved pancreatic viability.

Animals↗

Sp1 protein contributes to airway-specific rat MUC 2 mucin gene transcription.

We have shown increases in the abundance of airway mucin mRNA during the pathogenesis of chronic obstructive pulmonary disease in rat models (Jany et al., 1991) and now seek to determine the underlying mechanisms. As transcriptional modulation may be involved, we provide here a functional analysis of the 5' flanking region of a rat mucin gene (MUC 2). Using deletion mutants to bp -859, we constructed expression cassettes in CAT vectors and transfected them into two MUC 2-expressing cell lines, SPOC 1, a rat airway epithelial cell line and IEC-6, a rat intestinal epithelial cell line, and into one MUC 2 non-expressing cell line, FR, a rat skin fibroblast cell line. Results indicated that nucleotides -59 to -40 mediated high level expression in SPOC 1, but not in the other cells. Used as a probe in gel shift assays, fragment -59/-40 formed complexes of differing mobilities when incubated with nuclear protein extracts from the three cell types. Mutation of the putative Sp1 binding site in the probe sequence interfered with protein binding in all three cell types, but anti-Sp1 antibody supershifted a band formed only by airway cell extracts. A model of airway cell-specific MUC 2 transcription is proposed.

Animals↗

Identification of the transcription termination site of the mouse nkx-1.2 gene: involvement of sequence-specific factors.

We have identified a transcription termination site in the 3' flanking region of the mouse nkx-1.2 gene. A downstream transcription regulatory element in the mouse nkx-1.2 gene was characterized by transferring its 3'-fragment into a chloramphenicol acetyl transferase (CAT) expression vector. Analysis of recombinant plasmids transfected into mouse NIH3T3 cells by CAT assay showed the possible region of regulation. There were two direct repeat structures containing poly(dG-dT) x poly(dC-dA) sequences (GT repeats) in this region. The precise location of transcription termination was mapped by nuclease S1 analysis of the transcripts from recombinant plasmids transfected into COSM6 cells. It was approximately 20 nucleotides upstream of the first GT repeat within the 5' sequences of the first element of the two direct repeats. Gel mobility shift assay and footprinting analysis demonstrated that nuclear DNA binding proteins bound specifically to the sequences where the termination occurred as well as the other sequences in the second element of the direct repeats. Southwestern analysis showed that 90-, 54-, 36- and 15-kDa nuclear proteins bound to the region of the termination. It is possible that one or more of those proteins are involved in blocking the elongation of the mouse nkx-1.2 gene transcript and then result in termination.

3T3 Cells↗

Separation of DNA sequencing fragments up to 1000 bases by using poly(ethylene oxide)-filled capillary electrophoresis.

We have demonstrated that DNA bases up to 1000 base pairs (bp) in a sequencing ladder can be separated using poly(ethylene oxide)-filled capillary electrophoresis (resolution of raw data = 0.5 at 966 bp). Separation performance of this sieving matrix has been tested under different experimental conditions. It was found that the electric field strength played a critical role in the onset of reptation and thus the separation efficiency. Optimized gel composition and concentration is required for good separation, but the total gel concentration should lie between 2.5 and 3.0%. We observed that the capillary length influences the number of theoretical plates and the maximum readable length of DNA. For sequencing up to 500 bp, relatively nonviscous solutions can be used, greatly facilitating the replacement of the sieving matrix in between runs.

DNA↗

Characterization of the gene encoding catechol 2,3-dioxygenase from Achromobacter xylosoxidans KF701.

Catechol 2,3-dioxygenase (C23O) catalyzes a meta cleavage of the aromatic ring in catechol to form 2-hydroxymuconic semialdehyde. A C23O gene was cloned from chromosomal DNA of A. xylosoxidans KF701, a soil bacterium degrading biphenyl, and expressed in E. coli HB101. In substrate specificity to catechol and its analogs, the C23O exhibited the highest aromatic ring-fission activity to catechol, and its relative activity to other dihydroxylated aromatics was 4-chlorocatechol > 4-methylcatechol > 3-methylcatechol >> 2, 3-dihydroxybiphenyl. Aromatic ring-fission activity of the C23O to catechol was about 40-fold higher than that to 2,3-dihydroxybiphenyl. Nucleotide sequence analysis of the C23O gene from A. xylosoxidans KF701 revealed an open reading frame consisting of 924 base pairs, and identified a putative ribosome-binding sequence (AGGTGA) at about 10 nucleotides upstream from the initiation codon. The open reading frame can encode a polypeptide chain with molecular weight of 34 kDa containing 307 amino acid residues. The deduced amino acid sequence of the C23O exhibited the highest homology with that of C23O from Pseudomonas sp. IC with 96% identity, and the least homology with that of C23O from P. putida F1 with 22% identity among reported C23O sequences. Furthermore, comparison of the C23O sequence with other extradiol dioxygenases has led to identification of evolutionally conserved amino acid residues whose possible catalytic and structural roles are proposed.

Alcaligenes↗

Cloning and characterization of the 5' flanking region of human ATP-citrate lyase gene.

Two phage clones, lambda hgACL21 and lambda hgACL28, harboring the 5' flanking region of human ATP-citrate lyase (ACL) gene were identified by screening about 1.5 X 10(6) recombinant plaques from the lambdaEMBL3-human placental genomic DNA library. The 5' flanking region of ACL had the CAAT box on -92 bp from the transcription initiation site (+1), however, the TATA box was not found. The primer extension and rapid amplification of cDNA end showed that mRNA is transcribed at a thymine extending 12 bp upstream of the reported cDNA end. The sequences of 5' flanking region in 1.5 kb size of human ACL showed 60% homology with those of rat; however, no homology was found in the exon 1 and intron 1 region. Several consensus sequences, including four Sp1 binding sites, were found in the 5' flanking region of this gene. The promoter activity was assayed by transfecting the 3' or 5' deletion clones of ACL-chloramphenicol acetyl transferase (CAT) plasmid into PLC/PRF5 cells. The clone that contains the part of the first intron sequences from -659 to +440 bp showed the highest CAT activity in the transient transfection assay. High promoter activities were maintained until the transcription initiation site was removed. It is suggested that the sequences from -213 to +12 which contain three Sp1-binding sequences, CAAT box, and the transcription initiation site were necessary as a mean of for exerting the basal promoter activity of ACL gene.

ATP Citrate (pro-S)-Lyase↗

Localization and sequence analysis of the phnH gene encoding 2-hydroxypent-2,4-dienoate hydratase in Pseudomonas sp. strain DJ77.

The phnDEFG genes of Pseudomonas sp. DJ77, which are responsible for the degradation of polyaromatic hydrocarbons and chlorinated aromatics, were located previously on the 6.8 kb XhoI fragment of chromosomal DNA. Here, we sequenced a downstream region hitherto unknown and identified the phnH gene encoding a 2-hydroxypent-2,4-dienoate hydratase, which is required for the conversion of 2-hydroxypent-2,4-dienoate to 4-hydroxy-2-oxovalerate in the meta-cleavage pathway of catechols. The relative position of the hydratase gene in the phn operon is unique compared to the other meta-cleavage operons which have a dehydrogenative branch of the pathway. The PhnH hydratase contains 264 amino acids with a Mr of 28048. The deduced amino acid sequence of the PhnH enzyme is 60.9-31.6% identical to those of homologous enzymes encoded by the todG, bphE, cmtF, bphH, bphX1, xylJ, dmpE, cumE, MTCY03C7.20 and etbE genes.

Amino Acid Sequence↗

Mutagenesis of the positively charged conserved residues in the 5' exonuclease domain of Taq DNA polymerase.

Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in the polymerase chain reaction method. Taq DNA polymerase has a domain at the amino terminus (residue 1 to 290) that has a 5' exonuclease activity and a domain at the C-terminus that catalyzes polymerase reaction. Taq DNA polymerase is classified into the pol I family which is represented by E. coli DNA polymerase I. The alignment of amino acid sequences for the 5' exonuclease domains of the pol I family DNA polymerases shows six highly conserved sequences called motifs A to F. Motif C contains three positively charged residues such as 74Arg, 82Lys and 85Arg which might be involved in catalysis. In order to understand the function of those residues, they are mutagenized to alanine. The 5' exonucleolytic activities of those mutated 5' exonucleases decreased by 80 to 90%, thereby implying that three positively charged residues play certain roles in the 5' exonuclease catalysis.

Amino Acid Sequence↗

Phorbol myristate acetate-dependent association of protein kinase C alpha with phospholipase D1 in intact cells.

A phospholipase D1 (PLD1) was purified from rat brain by the use of antibody-coupled protein A Sepharose. We found that protein kinase C alp (PKCalpha) stimulated PLD1 activity in the presence of phorbol myristate acetate (PMA). PMA-dependent association of PKCalpha with PLD1 was verified in NIH-3T3 fibroblast cells, and COS7 cells transiently expressing PLD1 as well as in vitro suggesting that the activation of PLD1 resulted from direct association of PKCalpha with PLD1.

3T3 Cells↗

A phospholipase A2 kinetic and binding assay using phospholipid-coated hydrophobic beads.

A novel kinetic and membrane-binding assay for phospholipase A2 (PLA2) has been developed utilizing phospholipid-coated hydrophobic styrene-divinylbenzene beads (5.2 +/- 0.3 microm diameter). Phospholipids formed a stable monolayer film on styrene-divinylbenzene beads with average surface packing density of (1.3 +/- 0.2) x 10(-2) molecule/A2. Secretory PLA2 readily hydrolyzed 1-palmitoyl-2-[3H]-oleoyl-sn-glycero-3-phosphoglycerol coated on styrene-divinylbenzene beads which could be easily monitored by measuring the radioactivity of fatty acid released to solution in the presence of bovine serum albumin. For human cytosolic PLA2 with high specificity for sn-2 arachidonyl group, styrene-divinylbenzene beads coated with 1-stearoyl-2-[14C]-arachidonyl-sn-glycero-3-phosphocholine and dioleoylglycerol (7:3, mol/mol) were used as substrate. PLA2 activity was linearly proportional to the enzyme concentration in the range from 1 to 150 nM for human class II secretory PLA2 and from 1 to 20 nM for cytosolic PLA2; the specific activity was 1.6 and 1.7 micromol/min/mg, respectively. Finally, styrene-divinylbenzene beads coated with polymerized 1,2-bis[12-(lipoyloxy) dodecanoyl]-sn-glycero-3-phosphoglycerol were used to measure the membrane binding affinity of PLA2, which in conjunction with kinetic data provides important insights into how PLA2 interacts with membranes.

Animals↗

Twist-mediated activation of the NK-4 homeobox gene in the visceral mesoderm of Drosophila requires two distinct clusters of E-box regulatory elements.

NK-4, also called msh2 and tinman, encodes a homeodomain transcription factor that is required for the development of the dorsal mesoderm and its derivatives in the Drosophila embryo. Genetic analyses indicate that NK-4 resides downstream of the mesodermal determinant twist, which encodes a basic helix-loop-helix-type transcription factor. However, the regulation of NK-4 by twist remains poorly understood. Using expression assays in cultured cells and transgenic flies, we show that two distinct clusters of E-box regulatory sequences, present upstream of the NK-4 gene, mediate NK-4 expression in the visceral mesoderm. These elements are conserved between the Drosophila melanogaster and Drosophila virilis NK-4 genes and serve as binding sites for Twist (E1 cluster) and NK-4 (E2 cluster) proteins. In cultured cells, Twist and NK-4 binding results in activation of NK-4 gene expression. In transgenic animals, the E1 and E2 clusters are functionally connected, and both elements are required for NK-4 activation in cells of the visceral mesoderm and also for NK-4 repression in cells of the somatic musculature. These results demonstrate that NK-4 is a direct transcriptional target for Twist and its own gene product in visceral mesodermal cells, supporting the idea that twist and NK-4 function in the subdivision of the mesoderm during Drosophila embryogenesis.

Animals↗