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Biomedical subjects

Y Park

Publications and source records attributed to Y Park.

At least 19 recordsLinked to original sources
PubMed2006

Characterization of an O-methyltransferase from soybean.

O-methyltransferases (OMTs) catalyze the transfer of a methyl group from S-adenosine-L-methionine to a hydroxyl group of an acceptor molecule to form methyl ether derivatives and can modify the basic backbone of a secondary metabolite. A new O-methyltransferase, SOMT-9, was cloned from Glycine max and found to encode a protein whose molecular weight is 27-kDa. SOMT-9 was expressed as a GST-fusion protein in Escherichia coli and several compounds such as caffeic acid, esculetin, narigenin, kaempferol, quercetin, and luteolin were tested as putative substrates of SOMT-9. HPLC and NMR results showed that SOMT-9 transfers a methyl group to the 3'-OH group of substrates having ortho-hydroxyl groups. SOMT-9 showed the highest affinity for quercetin, suggesting that SOMT-9 uses a flavonoid as a substrate. Based on its molecular weight and substrate specificity, SOMT-9 belongs to a new class of OMT and is likely to be involved in the biosynthesis of isorhamnetin.

Amino Acid Sequence↗
PubMed2006

Sustained expression of Mect1-Maml2 is essential for tumor cell growth in salivary gland cancers carrying the t(11;19) translocation.

Mucoepidermoid (MEC) salivary gland tumors arise from a t(11;19) rearrangement which generates a fusion oncogene, Mect1-Maml2, that functions to activate CREB-responsive target genes. To determine if sustained expression of Mect1-Maml2 is required for tumor cell growth, we first showed that ectopic expression of Mect1-Maml2 in rat epithelial RK3E cells is tumorigenic in vivo in nude mice and that excised xenografts continue to express the fusion oncogene. We then generated a hairpin RNAi vector that selectively suppressed the fusion peptide and showed that ectopic expression in either parotid or pulmonary MEC tumor cell lines containing the t(11;19) rearrangement resulted in at least 90% colony growth inhibition. In contrast, single nucleotide changes within this RNAi sequence abolished the ability to suppress Mect1-Maml2 protein and abolished all growth inhibition of these MEC tumor lines. In addition, the RNAi-specific vector had no effect on colony growth of non-MEC tumors including a lung tumor or two other salivary gland cell lines that do not express Mect1-Maml2. We also generated a mutant Mect1-Maml2 expression plasmid that carried silent nucleotide changes within the RNAi target sequence and observed that co-transfection of this mutant, but not wild-type Mect1-Maml2, could partially rescue RNAi growth inhibition in the MEC tumor line. The recent detection of acquired fusion oncogenes in epithelial solid tumors has suggested new possibilities for the diagnosis and therapy of these cancers. Our data show that the 'gain-of-function' activity from aberrant Mect1-Maml2 expression is a candidate therapeutic target for this group of malignant salivary gland tumors.

Base Sequence↗
PubMed2006

piggyBac-like elements in the tobacco budworm, Heliothis virescens (Fabricius).

We identified two different groups of piggyBac-like elements (PLE) in the tobacco budworm, Heliothis virescens, and named them HvPLE1 and HvPLE2. An intact copy of HvPLE1 revealed the characteristics of PLE: inverted terminal repeats, inverted subterminal repeats, and an open reading frame encoding transposase, whereas other HvPLE1 copies and all the HvPLE2 copies carried disruptive mutations in the region encoding transposase. We also identified none to two bands per genome hybridized to a probe of Trichoplusia ni piggyBac in genomic Southern blotting, which are different from HvPLE1 or HvPLE2. Analysis of the sequences of multiple copies of HvPLE1 and HvPLE2 suggests that the PLEs are closely related to the T. ni piggyBac, of relatively young age, and independently entered the H. virescens genome.

Amino Acid Sequence↗
PubMed2006

Cloning and characterization of protease inhibitor-like cDNAs from the Hessian fly mayetiola destructor (SAY).

Analysis of transcriptomes from the salivary glands and midgut of Hessian fly larvae Mayetiola destructor (say) identified a set of diverse cDNAs that encode proteins with a relatively high percentage (over 10%) of cysteinyl residues. Structural comparison of these putative proteins with known sequences in GenBank revealed that the positions of the cysteinyl residues in the identified proteins were highly conserved within a family of proteinase inhibitors despite very little overall sequence similarity. Phylogenetic analysis sorted this set of cDNAs into five different groups. To determine if these cDNAs indeed encode proteinase inhibitors, recombinant proteins were generated with two cDNAs from two different groups. Biochemical analysis of the recombinant proteins against commercial and insect gut proteinases demonstrated that the recombinant proteins are strong proteinase inhibitors with different specificities. Northern blot and real-time PCR analysis revealed that the different genes were expressed at different developmental stages and in different tissues. The overall results indicated that M. destructor contains a complex of genes that code for proteinase inhibitors which may regulate proteinase activities in different regulatory pathways. The GenBank accession numbers for the cDNAs in this paper were DQ232690 to DQ232718.

Amino Acid Sequence↗
PubMed2006

Intact mariner-like element in tobacco budworm, Heliothis virescens (Lepidoptera: Noctuidae).

An intact mariner-like element was isolated from the genome of tobacco budworm, Heliothis virescens. This is the first report of an intact mariner element after the initial identification of Mos1 from Drosophila mauritiana. The full-length Hvmar1 has 30 bp inverted terminal repeats and a complete 1065 bp open reading frame (ORF) encoding the transposase with a 'D,D(34)D' motif in the catalytic domain. Polymerase chain reaction results show that at least one insertion of the Hvmar1 element is conserved in this Heliothis strain. Phylogenetic analysis demonstrated that Hvmar1 belongs to the irritans subfamily.

Amino Acid Sequence↗
PubMed2005

BMP-2 induces the expression of chondrocyte-specific genes in bovine synovium-derived progenitor cells cultured in three-dimensional alginate hydrogel.

OBJECTIVE: According to recent reports, the synovial membrane may contain mesenchymal stem cells with the potential to differentiate into chondrocytes under appropriate conditions. In order to assess the usefulness of synovium-derived progenitor cells for the purposes of cartilage tissue engineering, we explored their requirements for the expression of chondrocyte-specific genes after expansion in vitro. DESIGN: Mesenchymal progenitor cells were isolated from the synovial membranes of bovine shoulder joints and expanded in two-dimensions on plastic surfaces. They were then seeded either as micromass cultures or as single cells within alginate gels, which were cultured in serum-free medium. Under these three-dimensional conditions, chondrogenesis is known to be supported and maintained. Cell cultures were exposed either to bone morphogenetic protein-2 (BMP-2) or to isoforms of transforming growth factor-beta (TGF-beta). The levels of mRNA for Sox9, collagen types I and II and aggrecan were determined by RT-PCR. RESULTS: When transferred to alginate gel cultures, the fibroblast-like synovial cells assumed a rounded form. BMP-2, but not isoforms of TGF-beta, stimulated, in a dose-dependent manner, the production of messenger RNAs (mRNAs) for Sox9, type II collagen and aggrecan. Under optimal conditions, the expression levels of cartilage-specific genes were comparable to those within cultured articular cartilage chondrocytes. However, in contrast to cultured articular cartilage chondrocytes, synovial cells exposed to BMP-2 continued to express the mRNA for alpha1(I) collagen. CONCLUSIONS: This study demonstrates that bovine synovium-derived mesenchymal progenitor cells can be induced to express chondrocyte-specific genes. However, the differentiation process is not complete under the chosen conditions. The stimulation conditions required for full transformation must now be delineated.

Alginates↗
PubMed2005

Inactivated mariner-like elements (MLE) in pink bollworm, Pectinophora gossypiella.

We isolated multiple copies of mariner-like element (MLE) from the pink bollworm (PBW), Pectinophora gossypiella, a key lepidopteran cotton pest. Although all the MLE sequences contain multiple mutations accumulated in their transposase coding region, the consensus sequence revealed a putative ancestral transposase encoding 339 amino acid residues with a D,D(34)D motif, and 36 bp inverted terminal repeats, belonging to the cecropia subfamily, and most similar to the MLE found in Antheraea species. Examining six different pink bollworm populations, we conclude that the MLE in PBW described in this study are ancient and are undergoing the process of accumulating inactivating mutations. This conclusion is supported by the patterns of polymorphisms revealed by genomic Southern hybridization, transposable element displays, and sequences from multiple MLE.

Amino Acid Sequence↗
PubMed2004

Comparative mapping of ZYMV resistances in cucumber (Cucumis sativus L.) and melon (Cucumis melo L.).

Zucchini yellow mosaic virus (ZYMV) routinely causes significant losses in cucumber ( Cucumis sativus L.) and melon ( Cucumis melo L.). ZYMV resistances from the cucumber population 'TMG1' and the melon plant introduction (PI) 414723 show different modes of inheritance and their genetic relationships are unknown. We used molecular markers tightly linked to ZYMV resistances from cucumber and melon for comparative mapping. A 5-kb genomic region (YCZ-5) cosegregating with the zym locus of cucumber was cloned and sequenced to reveal single nucleotide polymorphisms and indels distinguishing alleles from ZYMV-resistant (TMG1) and susceptible (Straight 8) cucumbers. A low-copy region of the YCZ-5 clone was hybridized to bacterial artificial chromosome (BAC) clones of melon and a 180-kb contig assembled. One end of this melon contig was mapped in cucumber and cosegregated with ZYMV resistance, demonstrating that physically linked regions in melon show genetic linkage in cucumber. However the YCZ-5 region segregated independently of ZYMV resistance loci in two melon families. These results establish that these sources of ZYMV resistances from cucumber TMG1 and melon PI414723 are likely non-syntenic.

Chromosome Mapping↗
PubMed2004

Prediction of the risk of type 1 diabetes from polymorphisms in candidate genes.

Type 1 diabetes (T1D) is a T-cell-mediated autoimmune disease, in which pancreatic beta cells are selectively destroyed. Innate immune response is also important in the development of T1D. Several studies have demonstrated that HLA class II alleles, DQ and DR influence T1D susceptibility. Specific class I and II alleles are non-randomly associated with each other on an extended haplotypes, the typing of which provide the best risk determinants of T1D. Studying the haplotype in different ethnic populations will enable us to identify the exact polymorphisms that can trigger T1D and to develop biological tools for protection. With the advent of numerous candidate markers, additional loci that influence susceptibility to T1D have been reported. Although different studies have suggested that various genetic variants increase the risk, large-scale association studies that examine many polymorphisms simultaneously are required to allow reliable prediction of the genetic risk. Not only the numbers of genetic markers we are applying, but also the accurate phenotyping is critical in the success of setting-up a best-fitting model for prediction. For a better phenotyping, multiple autoantibodies to islet cell antigens, which may arise even before the clinical onset have been known as the best surrogate markers of T1D. However, differences in genotypes between antibody-positive individuals and T1D patients may indicate genetic factors, which determine progression to clinical disease. The ability to detect those antibody-positive individuals who will progress to T1D is central to prevention program.

Diabetes Mellitus, Type 1↗
PubMed2004

An improved large-scale synthesis of PEG-peptides for gene delivery.

Polyethylene glycol (PEG)-peptides are under development as components of nonviral gene delivery systems. Several earlier reports have demonstrated that covalent attachment of PEG to the surface of peptide condensed DNA particles blocks non-specific biodistribution during gene targeting. In this study, we report an improved large-scale synthesis and purification of a DNA condensing PEG-peptide used for gene delivery. The new method takes advantage of low-pressure cation-exchange chromatography to isolate dimeric Cys-Trp-Lys(18). The dimeric peptide was reduced and directly conjugated with PEG-maleimide resulting in PEG-Cys-Trp-Lys(18). The PEG-peptide was purified by low-pressure chromatography affording 50 mumol (400 mg) quantities of PEG-peptide in >95% purity. The approach offers the advantage of avoiding preparative high-performance liquid chromatography (HPLC) purifications of polylysine peptides to increase yield and capacity.

Dimerization↗
PubMed2004

Benzodiazepine impairment of perirhinal cortical plasticity and recognition memory.

Benzodiazepines, including lorazepam, are widely used in human medicine as anxiolytics or sedatives, and at higher doses can produce amnesia. Here we demonstrate that in rats lorazepam impairs both recognition memory and synaptic plastic processes (long-term depression and long-term potentiation). Both impairments are produced by actions in perirhinal cortex. The findings thus establish a mechanism by means of which benzodiazepines impair recognition memory. The findings also strengthen the hypotheses that the familiarity discrimination component of recognition memory is dependent on reductions in perirhinal neuronal responses when stimuli are repeated and that these response reductions are due to a plastic mechanism also used in long-term depression.

Animals↗
PubMed2004

Inhibition of adherence and killing of Candida albicans with a 23-Mer peptide (Fn/23) with dual antifungal properties.

Candida albicans adheres to host tissue and then proliferates in order to establish a commensal as well as a pathogenic state. Specific adherence to proteins is provided by several surface adhesins of Candida. Two well-studied proteins, Als1p and Als5p, do not require energy for adherence to occur (dead as well as living cells adhere) and have a multiplier effect of cell-cell aggregation that mediates the formation of microcolonies of Candida cells. The entire process is spontaneous, reversible, and stable for physiologically relevant chemical and physical forces. This adherence process is inhibited by the addition of free peptide ligands, including a 23-mer derived from fibronectin (Fn/23) that binds to the adhesins through H bond formation. Adherence was measured by determining the number of yeast cells that adhered to 90-microm-diameter polyethylene glycol (PEG) beads with a 7-mer peptide (KLRIPSV) synthesized on the surfaces of the beads. The concentration of the Fn/23 peptide that inhibited the adherence of cells to the peptide-coated beads by 50% was 4 to 5 microM, and the magnitudes of adherence were similar regardless of the presence or absence of physiologic salt concentrations. The minimum fungicidal concentration of Fn/23 was 2 to 4 microM in water, but there was no killing in physiologic salt concentrations. Peptides from the C and N termini or the center sequence of Fn/23 had no effect on inhibition of adherence and little effect on fungal viability. The fungicidal effect was similar to that seen with 23-, 19-, and 18-mer peptides derived from porcine myeloid cells, a Helicobacter pylori ribosomal protein, and a hybrid of cecropin and magainin, respectively. However, these fungicidal peptides did not inhibit C. albicans adherence to the peptide-coated PEG beads. This dual property of Fn/23, i.e., inhibition of adherence and killing of C. albicans, may provide important adjuvant effects in the treatment of disease caused by this fungus.

Amino Acid Sequence↗
PubMed2003

A dominant negative PPARgamma mutant shows altered cofactor recruitment and inhibits adipogenesis in 3T3-L1 cells.

AIMS/HYPOTHESIS: PPARgamma, a member of the nuclear hormone receptor family of transcription factors, plays a key role in adipocyte differentiation and insulin sensitivity. The aim of this study was to identify a potential dominant negative murine PPARgamma mutant and to characterize the in vitro functional properties of this mutant. METHODS: In vitro transient transfections and mammalian two-hybrid assays in TSA201 cells were used to characterize the transcriptional activity of the L466A mutant and to study the molecular interaction of transcriptional cofactors with the L466A mutant in an attempt to elucidate the mechanism of its dominant negative activity. Adenoviral constructs expressing PPARgamma wild-type (AdWT) or the L466A mutant (AdL466A) were infected into the murine 3T3-L1 cell line to study the mutant's effect on adipogenesis. RESULTS: The L466A mutant alone is transcriptionally defective. However, it retains DNA binding and inhibits the ligand-dependent and -independent activity of the wild-type receptor, consistent with dominant negative properties. In mammalian two-hybrid studies, the L466A mutant does not bind nuclear receptor coactivators. However, it more avidly recruits corepressors due to enhanced binding to the corepressor ID1 domain, leading to pronounced transcriptional repression. The AdL466A mutant inhibits adipogenesis induced by either a differentiation cocktail or by thiazolidinedione ligand. AdL466A infection also blocked the upregulation of the adipocyte marker genes aP2 and adipsin. CONCLUSION: We conclude that the L466A PPARgamma mutant possesses potent dominant negative activity based on preferential corepressor recruitment and it inhibits adipogenesis and the expression of adipocyte-specific genes.

3T3 Cells↗
PubMed2003

A rapid immunochromatographic assay for hepatitis B virus screening.

Simple, rapid and accurate assays for hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) are helpful for clinical diagnosis and field epidemiological surveys. A commercially developed, rapid immunochromatographic test for simultaneous detection of HBsAg and HBeAg was evaluated using a total of 2463 selected samples (827 frozen sera, 1011 fresh sera, and 625 whole blood samples). Results of the rapid test were compared with standard enzyme immunoassay (EIA) methods for HBsAg and HBeAg detection. The accuracy of the rapid test was excellent and was similar for frozen sera, fresh sera and whole blood. The overall sensitivity and specificity for the detection of HBsAg were 95 and 100%, and the corresponding positive and negative predictive values were 100 and 99.7%, respectively. The sensitivity and specificity for the detection of HBeAg were slightly less than that for HBsAg, and were 80 and 98%, with positive and negative predictive values of 91 and 94%, respectively. Thus, compared with the EIA method, the rapid test was highly sensitive and accurate for the detection of HBsAg although somewhat less sensitive and specific for detection of HBeAg. Because of its speed, simplicity and flexibility, the rapid test is ideally suited for HBsAg and HBeAg screening in population-based epidemiological studies and in low risk populations, particularly in regions of the world where hepatitis B is endemic.

Chromatography↗
PubMed2003

Conservation of ecdysis-triggering hormone signalling in insects.

Pre-ecdysis- and ecdysis-triggering hormones (PETH and ETH) from endocrine Inka cells initiate ecdysis in moths and Drosophila through direct actions on the central nervous system (CNS). Using immunohistochemistry, we found Inka cells in representatives of all major insect orders. In most insects, Inka cells are numerous, small and scattered throughout the tracheal system. Only some higher holometabolous insects exhibit 8-9 pairs of large Inka cells attached to tracheae in each prothoracic and abdominal segment. The number and morphology of Inka cells can be very variable even in the same individuals or related insects, but all produce peptide hormones that are completely released at each ecdysis. Injection of tracheal extracts prepared from representatives of several insect orders induces pre-ecdysis and ecdysis behaviours in pharate larvae of Bombyx, indicating functional similarity of these peptides. We isolated several PETH-immunoreactive peptides from tracheal extracts of the cockroach Nauphoeta cinerea and the bug Pyrrhocoris apterus and identified the gene encoding two putative ETHs in the mosquito Anopheles gambiae. Inka cells also are stained with antisera to myomodulin, FMRFamide and other peptides sharing RXamide carboxyl termini. However, our enzyme immunoassays show that these antisera cross-react with PETH and ETH. Our results suggest that Inka cells of different insects produce only peptide hormones closely related to PETH and ETH, which are essential endocrine factors required for activation of the ecdysis behavioural sequence.

Animals↗
PubMed2002

Robustness of the in-degree exponent for the World-Wide Web.

We consider a stochastic model for directed scale-free networks following power laws in the degree distributions in both incoming and outgoing directions. In our model, the number of vertices grow geometrically with time with a growth rate p. At each time step, (i) each newly introduced vertex is connected to a constant number of already existing vertices with the probability linearly proportional to in-degree distribution of a selected vertex, and (ii) each existing vertex updates its outgoing edges through a stochastic multiplicative process with mean growth rate of outgoing edges g and its variance sigma(2). Using both analytic treatment and numerical simulations, we show that while the out-degree exponent gamma(out) depends on the parameters, the in-degree exponent gamma(in) has two distinct values, gamma(in)=2 for p>g and 1 for p<g, independent of different parameters values. The latter case has logarithmic correction to the power law. Since the vertex growth rate p is larger than the degree growth rate g for the World-Wide Web (WWW) nowadays, the in-degree exponent appears robust as gamma(in)=2 for the WWW.

Journal Article↗