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J H Ha

Publications and source records attributed to J H Ha.

At least 19 recordsLinked to original sources

The complete genome sequence of pepper severe mosaic virus and comparison with other potyviruses.

The complete nucleotide sequence of pepper severe mosaic virus (PepSMV) was determined. The viral genome consisted of 9890 nucleotides, excluding a poly (A) tract at the 3' end of the genome. The PepSMV RNA genome encoded a single polyprotein of 3085 amino acid residues, resulting in ten functionally distinct potyviral proteins. The lengths of the 5' nontranslated region (NTR) and the 3' NTR were 164 and 468 nucleotides, respectively. The genome organization of the virus was typical for members of the genus Potyvirus in the family Potyviridae. The coat protein amino acid sequence identity between PepSMV and the other 45 potyviruses ranged from 53.4 to 79.7%. Sequence alignments and phylogenetic analyses of the potyviral polyprotein sequences revealed that PepSMV was the closest to potato virus Y (PVY) and closely related to members of the PVY subgroup. Our genome sequence data clearly confirmed that PepSMV belongs to a separate species in the genus Potyvirus.

3' Untranslated Regions↗

Cactus mild mottle virus is a new cactus-infecting tobamovirus.

A new cactus-infecting tobamovirus, Cactus mild mottle virus (CMMoV), was isolated from diseased grafted cactus, Gymnocalycium mihanovichii and its molecular properties were characterized. CMMoV is distantly related to known species of the genus Tobamovirus on the basis of serological and sequence analyses. Western blot analysis showed that CMMoV is serologically unrelated to Sammon's Opuntia virus, which is the only known species of the genus Tobamovirus found in cactus plants. The 3'-terminal 2,910 nucleotides of CMMoV have been sequenced. The coat protein (CP) and movement protein (MP) genes encode 161 and 306 amino acids residues, respectively, and the 3' untranslated region (UTR) consists of 229 nucleotides long. The nucleotide and amino acid sequences of the CP of CMMoV were 39.6% to 49.2% and 25.8% to 40.3% identical to other seventeen tobamoviruses, respectively. The MP shared 34.9% to 40.6% and 16.3% to 27.0% and 44.6% to 63.4% identities, respectively, at the amino acid and nucleotide levels with other members of the genus. Percentage identities of nucleotides of the 3' UTR ranged from 42.5% to 63.4%. Phylogenetic tree analyses of the CP and MP suggest the existence of the fifth cactus-infecting subgroup in the genus Tobamovirus. Sequence analyses of these two viral proteins revealed that the highest amino acid sequence identity between the virus and seventeen other tobamoviruses was 40.6%, supporting the view that CMMoV is a new definite species of the genus Tobamovirus.

Cactaceae↗

Genome structure and complete sequence of genomic RNA of Daphne virus S.

The complete genomic nucleotide sequence and structure of Daphne virus S (DVS), a daphne-infecting member of the genus Carlavirus, were determined. The genome of DVS was 8,739 nucleotides long, excluding the poly (A) tails. The genome of DVS contained six open reading frames coding for proteins of Mr 227 kDa (viral replicase), 25 kDa, 11 kDa and 7 kDa (triple gene block TGB) proteins 1, 2 and 3), 35 kDa (coat protein; CP), and 12 kDa from the 5' to 3' ends; respectively. This is the typical genome structure of members of the genus Carlavirus. Overall amino acid sequence similarities for the six ORFs of DVS were from 58.5% to 13.2% to those of the other carlaviruses. The 227 kDa replicase of DVS shared 45.5-39.2% amino acid similarities to that of 8 other known carlaviruses. Results from phylogenetic analyses of viral replicases and CPs demonstrated that DVS is a close relative of Helenium virus S and Chrysanthemum virus B. A total of 13 isolates of DVS shared 100-95.9% identities for the amino acid level and 99.5-81.0% identities for the nucleotide level. This is the first report of the complete genome sequence and structure of DVS and supports the conclusion that DVS is a typical species of the genus Carlavirus.

Base Sequence↗

Determination of the subthreshold state contribution in 13C(alpha,n)16O, the main neutron-source reaction for the s process.

The reaction rate of the stellar reaction 13C(alpha,n)16O, which is currently considered to be the main neutron source for the slow (s) process at low energies, has been rederived using the direct alpha-transfer reaction 13C(6Li,d)17O leading to the subthreshold state at 6.356 MeV in 17O. The contribution of the subthreshold state is found to be much smaller than the currently accepted predictions for the main neutron source of the s process, indicating less of a role of this reaction as the neutron source for the s-process scenario in low-mass stars at the asymptotic giant branch.

Journal Article↗

Zantedeschia mosaic virus causing leaf mosaic symptom in calla lily is a new potyvirus.

Anovel virus, Zantedeschia mosaic virus (ZaMV-KR), causing mosaic and malformation symptoms was isolated from calla lily ( Zantedeschia spp.) in Korea and its biological and molecular properties were characterized. The virus was distinct from Dasheen mosaic virus, an Araceae-infecting potyvirus, by serological and sequence analyses. Multiple alignments of the CP amino acid sequence between the virus and other potyviruses showed 51.8 to 62.1% identity. Phylogenetic analyses of the CP revealed that the virus could be clustered with Plum pox virus and Turnip mosaic virus. Sequence comparison of the CP gene between the virus and three other ZaMV isolates from Taiwan showed over 93.9% identity, and most of amino acids changes occurred in the N-terminal region. Sequence comparison of 3' NTR revealed homology levels of 27.0 to 47.9% between the virus and other potyviruses. Our results support ZaMV as a distinct species of the genus Potyvirus.

3' Untranslated Regions↗

In vitro effects of hydroxybenzaldehydes from Gastrodia elata and their analogues on GABAergic neurotransmission, and a structure-activity correlation.

The present study was designed to characterize the modulatory effects of the constituents of Gastrodia elata and their analogues on the GABAergic neurotransmission. 4-Hydroxybenzaldehyde (1) and 4-hydroxy-3-methoxybenzaldehyde (4) inhibited potently the activity of GABA transaminase (IC(50) = 4.1 and 5.4 microg/ml, respectively), while the activity of another constituent, 4-hydroxybenzyl alcohol (2), was very weak. Further investigation with 10 analogues revealed a structure-activity correlation, suggesting that the aldehyde group and the hydroxy group at C-4 are necessary for the inhibitory effect on the enzyme activity. Some potent enzyme inhibitors were examined for the effect on the radioligands to the GABA(A) receptor complexes of rat cerebral cortices. Among them, the component 4 dose-dependently increased (20 - 30 %) the binding of [(3)H]flunitrazepam in the presence of GABA.

4-Aminobutyrate Transaminase↗

Treatment of acute cerebral infarction with arginine esterase: a controlled study with heparin.

BACKGROUND AND PURPOSE: [corrected] There is no treatment proven to be of definitive benefit for ischemic stroke. Arginine esterase, a natural product from a snake venom, has been shown to reduce the serum fibrinogen level in human beings and may be useful in the treatment of ischemic stroke. In the present study, we compared the therapeutic effect of arginine esterase with that of heparin. SUBJECTS AND METHODS: We studied 50 consecutive patients with acute ischemic stroke who were admitted to the Asan Medical Center. We randomly administered either arginine esterase 0.005 unit/kg x 2 times/day or heparin (activated partial thromboplastin time 2-3 times of baseline value) intravenously for 7 days. Antiplatelets were administered afterwards in both groups. Blood fibrinogen, fibrinogen degradation product (FDP) and D-dimer levels were measured at 0, 6, 12, 18 h and 1, 2, 3, 7 and 30 days after the onset of stroke. NIH stroke scale was measured daily by 2 neurologists while Barthel index and Rankin scale were assessed at 7 days and 1 month after the onset of stroke by a research nurse. All these investigators were blinded to the therapeutic regimen each patient received. RESULTS: There were no significant differences in the mean age, gender proportion, stroke subtypes and baseline neurological severity between the two groups. One patient in the arginine esterase group died in an acute stage due to massive herniation and 1 in the heparin group underwent surgery for herniation. One (arginine esterase group) died of massive gastrointestinal bleeding due to previously unrecognized stomach cancer. Otherwise, no significant clinical and laboratory side effects were observed in both groups. In the arginine-esterase treated group, D-dimer and FDP levels were significantly (p < 0.05) elevated, and fibrinogen level significantly (p < 0.05) decreased at 2-7 days after the onset of stroke compared to the heparin-treated group. However, there was no significant difference in the neurological improvement reflected by NIH stroke scale, Barthel index and Rankin scale. CONCLUSION: Arginine esterase seems to be safe and has significant fibrinolytic effects when administered in the patients with acute ischemic stroke. However, in this preliminary study, it was not superior to heparin in terms of the improvement of neurological deficits. Further studies with larger doses and a larger number of subjects are required.

Acute Disease↗

Effects of angiotensin II on the renal antioxidant activities of borderline hypertensive rats.

This study was aimed to study the angiotensin II (Ang II)-induced antioxidant changes in the kidney of borderline-hypertensive rats (BHR). We measured renal antioxidant enzyme activities, and glutathione (GSH) contents and lipid peroxide levels in relation to the age of subjects. In the antioxidant enzyme assays, consistent changes were not observed in relation to age. However, in the assay for reduced GSH, nonenzymatic antioxidant, contents of adult and aged rats were much greater than those of weanling rats. Subcutaneous injection of pressor dose of human Ang II (200 microg/kg over 90 min) significantly reduced enzymatic activities in the weanling (4-week-aged) and adult (10-week-aged) BHR. However, in the relatively aged (16-week-aged) rats, Ang II did not alter enzymatic activities. Renal GSH contents of aged BHR, were highly increased by Ang II. Renal lipid peroxide levels of weanling and adult BHR were increased by Ang II, but decreased in the aged rats. However, these characteristic changes of renal antioxidant due to Ang II of the BHR could not be observed in the age-matched control, Wistar-Kyoto rats (WKR). From these results, it can be concluded that impacts of oxidative stress on the kidney of BHR may be greater in the young rats.

Aging↗

Suitability of AFLP markers for the study of genetic relationships among Korean native dogs.

To determine the genetic relationships among domestic dog breeds, we performed both a sequence comparison of mitochondrial DNA (mtDNA) and an amplified fragment length polymorphisms (AFLP) analysis. Three of four regions of mtDNA, cytochrome b, cytochrome oxidase subunit II, and 16S rRNA genes were highly homogeneous among dog breeds, whereas the other region, the control region, showed relatively high polymorphisms with a maximum percentage difference of 3.18%. However, the control region showed extensive polymorphism even within breeds, and the relationship tree derived from the data could not clearly delimit distinct breeds. 19 EcoRI/MseI primer combinations were used to generate AFLP markers among 25 dogs from 11 breeds including three Korean native dogs. These amplification reactions allowed the detection of more than 1900 amplification products of which 408 were identified as polymorphic bands. Unrooted neighbor-joining tree based on dissimilarity values showed that the Korean native dogs were clustered together with the Asian dogs and that the Asian originated dogs were clustered separately from Western originated dogs. A consensus tree using parsimony method also showed Korean native dogs were grouped separately from the other dogs with moderate bootstrap values. Taken together, it is concluded that AFLP analysis is a more informative tool for revealing genetic relationships among dog breeds than mtDNA sequence comparison.

Animals↗

Single amino acid of g(alpha16) (Ala(228)) is responsible for the ability of chemoattractant C5a receptor to induce G(alpha16)-mediated inositol phosphate release.

Our previous study suggested that the region encompassing residues 220-240 on G(alpha16) is important in coupling with C5a receptor (Lee et al. (1995) Mol. Pharmacol. 47, 218-223). When aligned sequences are compared in the residue 220-240 segment of G(alpha16), there is a block of eight amino acids extending from residue 227 to residue 234 (227-Ile-Ala-Leu-Ile-Tyr-Leu-Ala-Ser-234) in G(alpha16) that is replaced by a heterologous block extending from amino acid residue 224 to residue 231 (224-Thr-Ser-Ile-Met-Phe-Leu-Val-Ala-231) in G(alpha11). In order to identify the specific amino acid residue necessary for coupling to C5a receptor within the extension of eight amino acids in G(alpha16), a series of chimeric G(alpha11)/G(alpha16) cDNA constructs and mutant G(alpha16) cDNAs were expressed. Then the ability of chimeras and mutant proteins to mediate C5a-induced release of inositol phosphate in transfected Cos-7 cells was tested. The results show that single amino acid Ala(228) is responsible for conferring about 40-50% of the activity of G(alpha16) induced by C5a receptor stimulation.

Alanine↗

Cloning of human acetyl-CoA carboxylase beta promoter and its regulation by muscle regulatory factors.

The 280-kDa beta-isoform of acetyl-CoA carboxylase (ACCbeta) is predominantly expressed in heart and skeletal muscle, whereas the 265-kDa alpha-isoform (ACCalpha) is the major ACC in lipogenic tissues. The ACCbeta promoter showed myoblast-specific promoter activity and was strongly induced by MyoD in NIH3T3 cells. Serial deletions of the promoter revealed that MyoD acts on the E-boxes located at positions -498 to -403 and on the proximal region including the 5'-untranslated region. Destruction of the E-boxes at positions -498 to -403 by site-directed mutagenesis resulted in a significant decrease of MyoD responsiveness. The "TGAAA" at -32 to -28 and the region around the transcription start site play important roles in basal transcription, probably as a TATA box and an Inr element, respectively. Mutations of another E-box at -14 to -9 and a "GCCTGTCA" sequence at +17 to +24 drastically decreased the MyoD responsiveness. The novel cis-element GCCTGTCA was preferentially bound by MyoD homodimer in EMSA and conferred MyoD responsiveness to a luciferase reporter, which was repressed by the overexpression of E12. This finding is unique since activation via E-boxes is mediated by heterodimers of MyoD and E-proteins. We screened a human skeletal muscle cDNA library to isolate clones expressing proteins that bind to the region around the GCCTGTCA (+8 to +27) sequence, and isolated Myf4 and Myf6 cDNAs. Electrophoretic mobility shift assay showed that recombinant Myf4 and Myf6 bind to this novel cis-element. Moreover, transient expression of Myf6 induced significant activation on the ACCbeta promoter or an artificial promoter harboring this novel cis-element. These findings suggest that muscle regulatory factors, such as MyoD, Myf4, and Myf6, contribute to the muscle-specific expression of ACCbeta via E-boxes and the novel cis-element GCCTGTCA.

Acetyl-CoA Carboxylase↗

Genomic analysis and functional expression of canine dopamine D2 receptor.

Dopamine D2 receptor (DRD2) is one of the five dopamine receptors with seven transmembrane domains that are coupled to the G protein. We have cloned and characterized the genomic and cDNA sequences of the canine DRD2 gene, which are 12.7 and 2.7 kb in size, respectively. The genomic DNA is composed of seven exons and six introns, encoding a 443 amino acid protein with 95% amino acid identity to other mammalian D2 receptors. A length polymorphism was detected in intron 3 of the receptor gene. We also characterized alternatively spliced forms of DRD2 cDNAs, DRD2L and DRD2S. They showed a higher level of expression in midbrain and thalamus. The ratio between the long and short form is similar in RT-PCR reaction. In human and rodent, the same two spliced forms are known to be coupled to G(i)-type heterotrimeric GTP binding protein, thereby opening an inwardly rectifying potassium channel, GIRK1. When the canine DRD2L and DRD2S were heterologously expressed in Xenopus oocytes, both forms activated GIRK1 potassium channels through coupling with G(i) protein. This activation was dose-dependent, demonstrating its ligand specificity.

Amino Acid Sequence↗

4-Hydroxybenzaldehyde from Gastrodia elata B1. is active in the antioxidation and GABAergic neuromodulation of the rat brain.

Ether fraction of G. elata methanol extract significantly inhibited the recovery time and severity induced by pentylenetetrazole (PTZ) treatment. Pretreatment of ether fraction of G. elata methanol extract successfully prevented diminution of brain GABA level in subconvulsive dose of PTZ-treated rats. 4-Hydroxybenzaldehyde, an analogue of p-hydroxybenzyl alcohol, showed an inhibitory effect on the GABA transaminase, and its inhibitory activity was higher than that of valproic acid, a known anticonvulsant. In the brain of PTZ-treated rats, brain lipid peroxidation was significantly increased, while it recovered to the control level after treatment with 4-hydroxybenzaldehyde. It may be concluded that antioxidation and positive modulation of GABAergic neuromodulation of 4-hydroxybenzaldehyde partially contribute to an antiepileptic and anticonvulsive activity of G. elata B1.

4-Aminobutyrate Transaminase↗

Increase of [Ca(2+)]i and release of arachidonic acid via activation of M2 receptor coupled to Gi and rho proteins in oesophageal muscle.

We have previously shown that acetylcholine-induced contraction of oesophageal circular muscle depends on activation of phosphatidylcholine selective phospholipase C and D, which result in formation of diacylglycerol, and of phospholipase 2 which produces arachidonic acid. Diacylglycerol and arachidonic acid interact synergistically to activate protein kinase C. We have therefore investigated the relationship between cytosolic Ca(2+) and activation of phospholipase A(2) in response to acetylcholine-induced stimulation, by measuring the intracellular free Ca(2+) ([Ca(2+)]i), muscle tension, and [3H] arachidonic acid release. Acetylcholine-induced contraction was associated with increased [Ca(2+)]i and arachidonic acid release in a dose-dependent manner. In Ca(2+)-free medium, acetylcholine did not produce contraction, [Ca(2+)]i increase, and arachidonic acid release. In contrast, after depletion of Ca(2+) stores by thapsigargin (3 microM), acetylcholine caused a normal contraction, [Ca(2+)]i increase and arachidonic acid release. The increase in [Ca(2+)]i and arachidonic acid release were attenuated by the M2 receptor antagonist methoctramine, but not by the M3 receptor antagonist p-fluoro-hexahydro siladifenidol. Increase in [Ca(2+)]i and arachidonic acid release by acetylcholine were inhibited by pertussis toxin and C3 toxin. These findings indicate that contraction and arachidonic acid release are mediated through muscarinic M2 coupled to Gi or rho protein activation and Ca(2+) influx. Acetylcholine-induced contraction and the associated increase in [Ca(2+)]i and release of arachidonic acid were completely reduced by the combination treatment with a phospholipase A(2) inhibitor dimethyleicosadienoic acid and a phospholipase D inhibitor pCMB. They increased by the action of the inhibitor of diacylglycerol kinase R59949, whereas they decreased by a protein kinase C inhibitor chelerythrine. These data suggest that in oesophageal circular muscle acetylcholine-induced [Ca(2+)]i increase and arachidonic acid release are mediated through activation of M2 receptor coupled to Gi or rho protein, resulting in the activation of phospholipase A(2) and phospholipase D to activate protein kinase C.

Acetylcholine↗

Electrocardiographic abnormalities in patients treated with clozapine.

BACKGROUND: Cardiovascular side effects of clozapine are not uncommon, but few systematic studies of these effects have been performed. In this study, we reviewed data on the electrocardiographic (ECG) abnormalities in patients treated with clozapine. METHOD: Sixty-one patients treated with clozapine were selected from the Seoul National University Hospital Treatment-Resistant Schizophrenia Clinic. A retrospective chart review was conducted to identify ECG abnormalities and cardiovascular side effects. RESULTS: The prevalence of ECG abnormalities in patients who had been using antipsychotics other than clozapine was 13.6% at baseline, which increased significantly to 31.1% after commencement of clozapine treatment. Among the 53 patients without baseline ECG abnormalities, 13 showed new-onset ECG abnormalities after using clozapine. Normal ECG under previous antipsychotic medication reduced the risk of new-onset ECG abnormalities, whereas increased age was found to increase the risk. The occurrence of orthostatic hypotension or tachycardia was not related to the development of ECG abnormalities. Most of the newly developed abnormalities had little clinical significance, and they tended to occur during the initial phase of treatment. In 10 patients, ECGs normalized despite the continued use of clozapine. Clozapine increased corrected QT interval (QTc) in a dose-dependent fashion; however, the clinical significance of this observation is uncertain. Pathologic prolongation of QTc was found to be rare. CONCLUSION: Although a substantial portion of patients treated with clozapine developed ECG abnormalities, most of the abnormalities were benign and did not hinder further treatment.

Adolescent↗

Identifying the site of initial tertiary structure disruption during apomyoglobin unfolding.

Structural characterization of protein unfolding intermediates [Kiefhaber et al. (1995) Nature 375, 513; Hoeltzli et al.(1995) Proc. Natl. Acad. Sci. U.S.A. 92, 9318], which until recently were thought to be nonexistent, is beginning to give information on the mechanism of unfolding. To test for apomyoglobin unfolding intermediates, we monitored kinetics of urea-induced denaturation by stop-flow tryptophan fluorescence and quench-flow amide hydrogen exchange. Both measurements yield a single, measurable kinetic phase of identical rate, indicating that the reaction is highly cooperative. A burst phase in fluorescence, however, suggests that an intermediate is rapidly formed. To structurally characterize it, we carried out stop-flow thiol-disulfide exchange studies of 10 single cysteine-containing mutants. Cysteine probes buried at major sites of helix-helix pairing revealed that side chains throughout the protein unpack and become accessible to the labeling reagent [5, 5'-dithiobis (2-nitrobenzoic acid)] with one of two rates. Probes located at all helical-packing interfaces-except for one-become exposed at the rate of global unfolding as determined by fluorescence and hydrogen exchange measurements. In contrast, probes located at the A-E helical interface undergo complete thiol-disulfide exchange within the mixing dead time of 6 ms. These results point to the existence of a burst-phase unfolding intermediate that contains globally intact hydrogen bonds but locally disrupted side-chain packing interactions. Dissolution of secondary and tertiary structure are therefore not tightly coupled processes. We suggest that disruption of tertiary structure may be a stepwise process that begins at the weakest point of the native fold, as determined by native-state hydrogen-exchange parameters.

Animals↗