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Biomedical subjects

J Y Yang

Publications and source records attributed to J Y Yang.

At least 37 records · Page 2Linked to original sources

Cloning, characterization and genomic organization of LCC-1 (scya16), a novel human CC chemokine expressed in liver.

By homology search of expressed sequence tags (EST) in GenBank a novel member of the CC chemokine family was identified. The full-length sequence of this liver-specific CC chemokine (LCC-1) predicted a mature protein of 97 amino acids with 31-48% identity to other CC chemokines. There was a characteristic amino acid C-term extension when aligned with other chemokines. Northern blot analysis from a panel of human tissues revealed that LCC-1 mRNA expression is restricted to adult and fetal liver. Different polyadenylation results in two mRNA species of 1.5 kb and 0.5 kb in size. LCC-1 is constitutively expressed in human HepG2 hepatoma cells and is induced by hypoxic exposure. The promoter region of the LCC-1 gene contains potential HIF-1 binding sites. The EST for LCC-1 has been previously mapped to the CC chemokine cluster on human chromosome 17q11.2. The organization of the LCC-1 gene (scya16) into three exons interrupted by two introns is identical to that found for other members of the CC chemokine family.

Adult↗

Preliminary X-ray crystallographic and NMR studies on the exonuclease domain of the epsilon subunit of Escherichia coli DNA polymerase III.

The structured core of the N-terminal 3'-5' exonuclease domain of epsilon, the proofreading subunit of Escherichia coli DNA polymerase III, was defined by multidimensional NMR experiments with uniformly (15)N-labeled protein: it comprises residues between Ile-4 and Gln-181. A 185-residue fragment, termed epsilon(1-185), was crystallized by the hanging drop vapor diffusion method in the presence of thymidine-5'-monophosphate, a product inhibitor, and Mn(2+) at pH 5.8. The crystals are tetragonal, with typical dimensions 0.2 mm x 0.2 mm x 1.0 mm, grow over about 2 weeks at 4 degrees C, and diffract X-rays to 2.0 A. The space group was determined to be P4(n)2(1)2 (n = 0, 1, 2, 3), with unit cell dimensions a = 60.8 A, c = 111.4 A.

Catalytic Domain↗

Large scalp and skull defect in aplasia cutis congenita.

Aplasia cutis congenita (ACC) is a rare disease, which may involve any site on the body but usually the vertex with scalp and even skull involvement. Superficial lesions may heal spontaneously and seldom result in morbidity or mortality. However, in patients with large scalp and skull defects, there are risks of infection and bleeding. Both surgical and conservative treatment have been proposed. We report a case of successful treatment of ACC by early debridement and wound closure with split-thickness skin graft (STSG) in the neonatal stage followed by replacement of the STSG with a hairy scalp flap by means of tissue expansion at the age of 3 years. The original skull defect, 7 x 10 cm in size, decreased to 1.2 x 1.2 cm after a 4.5-year follow-up. No complications, such as haemorrhage or meningitis, were encountered. The final result was satisfactory.

Ectodermal Dysplasia↗

NMR solution structure of the theta subunit of DNA polymerase III from Escherichia coli.

The catalytic core of Escherichia coli DNA polymerase III contains three tightly associated subunits (alpha, epsilon, and theta). The theta subunit is the smallest, but the least understood of the three. As a first step in a program aimed at understanding its function, the structure of the theta subunit has been determined by triple-resonance multidimensional NMR spectroscopy. Although only a small protein, theta was difficult to assign fully because approximately one-third of the protein is unstructured, and some sections of the remaining structured parts undergo intermediate intramolecular exchange. The secondary structure was deduced from the characteristic nuclear Overhauser effect patterns, the 3J(HN alpha) coupling constants and the consensus chemical shift index. The C-terminal third of the protein, which has many charged and hydrophilic amino acid residues, has no well-defined secondary structure and exists in a highly dynamic state. The N-terminal two-thirds has three helical segments (Gln10-Asp19, Glu38-Glu43, and His47-Glu54), one short extended segment (Pro34-Ala37), and a long loop (Ala20-Glu29), of which part may undergo intermediate conformational exchange. Solution of the three-dimensional structure by NMR techniques revealed that the helices fold in such a way that the surface of theta is bipolar, with one face of the protein containing most of the acidic residues and the other face containing most of the long chain basic residues. Preliminary chemical shift mapping experiments with a domain of the epsilon subunit have identified a loop region (Ala20-Glu29) in theta as the site of association with epsilon.

Amino Acid Sequence↗

Criss-crossed interactions between the enhancer and the att sites of phage Mu during DNA transposition.

A bipartite enhancer sequence (composed of the O1 and O2 operator sites) is essential for assembly of the functional tetramer of phage Mu transposase (MuA) on supercoiled DNA substrates. A three-site interaction (LER) between the left (L) and right (R) ends of Mu (att sites) and the enhancer (E) precedes tetramer assembly. We have dissected the role of the enhancer in tetramer assembly by using two transposase proteins that have a common att site specificity, but are distinct in their enhancer specificity. The activity of these proteins on substrates containing hybrid enhancers reveals a 'criss-crossed' pattern of interaction between att and enhancer sites. The left operator, O1, of the enhancer interacts specifically with the transposase subunit at the R1 site (within the right att sequence) that is responsible for cleaving the left end of Mu. The right operator, O2, shows a preferential interaction with the transposase subunit at the L1 site (within the left att sequence) that is responsible for cleaving the right end of Mu.

Attachment Sites, Microbiological↗

Change of plasma lipoproteins by heparin-released lipoprotein lipase.

Lipoprotein lipase (LPL) is known to be attached to the luminal surface of vascular endothelial cells in a complex with membrane-bound heparan sulfate, and released into blood stream by heparin. LPL that catalyzes hydrolysis of triglyceride (TGL) on chylomicron and VLDL into two fatty acids and monoacylglycerol, is also implicated to participate in an enhancement of cholesterol uptake by arterial endothelial cells in vitro. But little is known about the LPL-mediated cholesterol uptake in physiological state. In this study, changes in blood lipid composition and levels of lipoproteins were determined after the injection of heparin in human. The level of LPL in plasma was increased from 0 to 11 mU/ml within 30-40 min post-heparin administration and decreased to the basal level within 2 h. The level of TGL in plasma decreased from 70 mg/dl to 20 mg/dl within 1 h and gradually increased to 80 mg/dl within 4 h. However the level of total cholesterol in plasma remained at 140 mg/dl during an experimental period of 4 h. Analysis of Lipoproteins in plasma by NaBr density gradient ultracentrifugation showed that the level of VLDL decreased from 50 mg/dl to 10 mg/dl within 1-2 h and returned to normal plasm level at 4 h. However there were no significant changes in the level of LDL and HDL. These results suggest that, at least, in normo-lipidemic subjects, increased free plasm LPL acts primarily on VLDL and failed to show any significant uptake of cholesterol-rich lipoproteins in human.

Adult↗

Long-term efficacy of plasma-derived hepatitis B vaccine: a 15-year follow-up study among Chinese children.

To determine necessity and timing of booster of hepatitis B vaccine, we need to observe the duration of its protection. We report the results of a 15-year follow-up of a cohort of 649 children who participated a randomized, double blind, placebo-controlled trial on a plasma-derived hepatitis B vaccine in 1982. During the 15 years after vaccination, more vaccinated children had anti-HBs of 10 S/N ratios or over, compared with the controls, at all nine observations. At 15 years 50.0% (26/52) of the participants studied in the vaccinated group and 33.3% of the tested controls (18/54) retained anti-HBs levels of S/N ratios> or =10 (P < 0.09). However, since 5 years after vaccination, median S/N ratios of anti-HBs among the vaccinated children with detectable anti-HBs were lower than those of the controls except that detected at 15 years. 16.7% (9/54) of the tested children in the control group were HBsAg positive at 15 years after vaccination, in comparison with 1.9% (1/52) of the tested children in the vaccinated (P < 0.02). 28 chronic HBsAg carriers were identified in the control cohort over the 15 years, whereas only 1 case was noted in the vaccinated group (8.2% vs. 0.3%, P < 0.00001), corresponding to an efficacy of 96%.

Double-Blind Method↗

Heterozygous defect in HIV-1 coreceptor CCR5 and chemokine production.

CC chemokine receptor 5 (CCR5) is a cell entry cofactor for macrophage-tropic isolates of human immunodeficiency virus 1 (HIV-1). An inactive CCR5 allele with a 32-nucleotide deletion (CCR5Delta32) has been described that confers resistance to HIV-1 infection in homozygotes and slows the rate of progression to AIDS in heterozygotes. We found the allele CCR5Delta32 to be not rare in 399 Swiss blood donors with a frequency of 0.080. To assess the influence of defective CCR5 on production of its ligands we determined the capacity to produce the chemokines macrophage inflammatory protein (MIP)-1alpha, MIP-1beta and RANTES in comparison with the production of the CXC chemokine IL-8 which does not bind to CCR5. Production of chemokines was determined during endotoxin stimulation of whole-blood samples ex vivo. Both, basal and LPS-induced chemokine production in 32 blood donors heterozygous for CCR5Delta32 were not significantly different when compared with 55 blood donors who were homozygous for the wild type CCR5 allele.

Alleles↗

Levels of paraquat in fatal intoxications.

We describe here fatal levels of paraquat in plasma of victims due to the ingestion of the herbicide, paraquat, and the relationship between those levels and survival times after ingestion. We determined paraquat levels in plasma of 106 paraquat poisoning cases in Korea between June 1992 and December 1996 using a visible spectrophotometric method based on the alkali-dithionite reaction. Among 55 cases with known survival times, plasma paraquat levels of victims who died within one day of ingestion ranged from 2.3 to 636.6 microg/ml (average 127.6 microg/ml) while those of the victims who died between 1 approximately 4 days ranged from 0.9 to 25.1 microg/ml (average 7.0 microg/ml). Since preparations containing 24.5% paraquat dichloride are still used in Korea, our data for Korean victims who died within one day were 8.5 times higher than those in Baselt and Cravey's report for the same survival times. Our data also suggest that victims in whom a plasma paraquat concentration of more than 30 microg/ml was detected are likely to have died within 24 h after ingestion.

Adolescent↗

Forensic toxicological implication of acute fatal poisoning cases due to benfuracarb ingestion.

We describe here three cases involving acute fatalities due to benfuracarb ingestion and the forensic toxicological implications. Benfuracarb, a carbamate insecticide and its main metabolite carbofuran, were detected using thin layer chromatography (TLC) and gas chromatography/mass spectrophotometry (GC/MS) after extraction with ethyl acetate and then quantified using gas chromatography (GC) equipped with NPD. The blood levels of benfuracarb and carbofuran were in the range of 0.30-2.32 microg/ml and 1.45-1.47 microg/ml, respectively. Benfuracarb was not detected in urine, but carbofuran was detected in the range of 0.53-2.66 microg/ml.

Adult↗

Symptomatic tracheal stenosis in burns.

Tracheal stenosis in burns is rare and usually results from prolonged intubation or tracheostomy. Inhalation injury itself has the potential risk of tracheal stenosis. We reviewed the records of 1878 burn patients during 1987 to 1995 and found seven with tracheal stenosis (0.37%) after an average of 4.4 years follow up. There were 4 males and 3 females with an average age of 27.3 years. The tracheal stenosis developed 1-22 months after burn (average 7 months). Five patients had their inhalation injury confirmed by bronchoscopic examination. The incidence of tracheal stenosis among inhalation injury patients was 5.49% (5/92). Six patients needed intubation in the initial stage either for respiratory distress or prophylaxis, with an average duration of 195.2 h. In addition to prolonged intubation, the presence of inhalation injury, repeated intubations and severe neck scar contractures are also contributors to tracheal stenosis in burns. We favor T-tube insertion as the first treatment choice; permanent tracheostomy was unsatisfactory in our study.

Adolescent↗

Studies on the sedative and hypnotic effects of oleamide in mice.

The hypnotic and sedative effects of oleamide (CAS 301-02-0) were studied in mice. Intraperitoneal injection of oleamide showed a time-related inhibition of the locomotor activity with the maximum effect appearing 30 min after drug administration. Oleamide, at the dose range of 43.7-700 mg/kg, dose-dependently inhibited the locomotor activity in mice. Oleamide could promote the hypnotic action induced by sodium pentobarbital. The interaction studies showed that oleamide potentiated the inhibitory effect of diazepam (CAS 439-14-5) and antagonized the stimulatory effect of ethanol, methamphetamine, and caffeine, respectively. These results provide further evidence for the hypnotic and sedative effects of oleamide, suggesting a potential therapeutic usefulness of this fatty acid amide.

Animals↗

Dynamics of nontypical apoptotic morphological changes visualized by green fluorescent protein in living cells with infectious pancreatic necrosis virus infection.

Morphologically, apoptotic cells are characterized by highly condensed membrane blebbing and formation of apoptotic bodies. Recently, we reported that apoptosis precedes necrosis in a fish cell line infected with infectious pancreatic necrosis virus (IPNV). In the present study, we tested the possibility that nontypical apoptosis is a component of IPNV-induced fish cell death. A variant type of green fluorescent protein (EGFP) was expressed in a fish cell line such that EGFP served as a protein marker for visualizing dynamic apoptotic cell morphological changes and for tracing membrane integrity changes during IPNV infection. Direct morphological changes were visualized by fluorescence microscopy by EGFP in living cells infected with IPNV. The nontypical apoptotic morphological change stage occurred during the pre-late stage (6 to 7 h postinfection). Nontypical apoptotic features, including highly condensed membrane blebbing, occurred during the middle apoptotic stage. At the pre-late apoptotic stage, membrane vesicles quickly formed, blebbed, and were finally pinched off from the cell membrane. At the same time, at this pre-late apoptotic stage, apoptotic cells formed unique small holes in their membranes that ranged from 0.39 to 0.78 micrometer according to examination by scanning electron microscopy and immunoelectron microscopy. Quantitation of the intra- and extracellular release of EGFP by CHSE-214-EGFP cells after IPNV infection was done by Western blotting and fluorometry. Membrane integrity was quickly lost during the late apoptotic stage (after 8 h postinfection), and morphological change and membrane integrity loss could be prevented and blocked by treatment with apoptosis inhibitors such as cycloheximide, genistein, and EDTA before IPNV infection. Together, these findings show the apoptotic features at the onset of pathology in host cells (early and middle apoptotic stages), followed secondarily by nontypical apoptosis (pre-late apoptotic stage) and then by postapoptotic necrosis (late apoptotic stage), of a fish cell line. Our results demonstrate that nontypical apoptosis is a component of IPNV-induced fish cell death.

Animals↗

Assessment of myocardial viability using 123I-labeled iodophenylpentadecanoic acid at sustained low flow or after acute infarction and reperfusion.

UNLABELLED: 123I-labeled iodophenylpentadecanoic acid (IPPA) is a synthetic fatty acid that may be useful for determination of myocardial viability. We investigated the uptake and clearance kinetics of this tracer in canine models of ischemia and infarction. METHODS: In protocol 1, 185 MBq (5 mCi) 123I-IPPA were injected intravenously in 19 dogs with 50% left anterior descending artery (LAD) flow reduction. In 9 dogs, 201TI was coinjected. In protocol 2, 5 dogs underwent LAD occlusion for 3 h, and 123I-IPPA was injected 60 min after reperfusion. All dogs had flow measured by microspheres, regional systolic thickening by ultrasonic crystals and measurements of postmortem risk area and infarct size. Tracer activities were quantified by gamma well counting and by serial imaging. RESULTS: In protocol 1 dogs with sustained low flow (50% +/- 4%) and absence of systolic thickening (-3.2% +/- 1%), 123I-IPPA defect magnitude (LAD/left circumflex artery [LCX] count ratios) decreased from 0.65 +/- 0.02 to 0.74 +/- 0.02 at 30 min and to 0.84 +/- 0.03 at 2 h (P < 0.01), indicative of rest redistribution. Final transmural 123I-IPPA LAD/LCX activity ratio (0.99 +/- 0.05) was significantly greater than the flow ratio (0.53 +/- 0.04) at injection, confirming complete rest redistribution. The final 123I-IPPA activity ratio was significantly greater than the 201TI ratio over the 2-h period (P < 0.01). In protocol 2 dogs that underwent 3 h of total LAD occlusion and reflow (infarct size = 51% +/- 13% of risk area), viability was overestimated with 123I-IPPA, because uptake averaged 64% of normal in the central necrotic region, where flow averaged < 10% of normal. CONCLUSION: These findings suggest that serial 123I-IPPA imaging may be useful for assessing myocardial viability under conditions of sustained low flow and myocardial asynergy, such as appears to exist in patients with chronic coronary artery disease and depressed left ventricular function. In contrast, 123I-IPPA given early after reperfusion following prolonged coronary occlusion overestimates the degree of viability and therefore may not provide useful information pertaining to the degree of myocardial salvage after reflow in the setting of acute myocardial infarction.

Animals↗

Mutational analysis of domain II beta of bacteriophage Mu transposase: domains II alpha and II beta belong to different catalytic complementation groups.

This study examines the contribution of domain II beta of bacteriophage Mu transposase (A protein), a subdomain of the central catalytic domain II, to the transposition reaction. The properties of several point mutations implicate a role for this domain in facilitating metal-assisted assembly of the synaptic complex, as well as in intramolecular DNA strand transfer. Point mutations as well as deletions in domain II beta can be complemented by those in domain II alpha but not those in domain III alpha. Thus, residues within subdomains II alpha and II beta belong to different catalytic complementation groups.

Bacteriophage mu↗

Use of the scalp as a donor site for large burn wound coverage: review of 150 patients.

The characteristics of rapid wound healing and multiple harvest capacity make the scalp an important donor site when dealing with large and deep burn wounds. This paper reports the results of a retrospective analysis of 150 patients treated for large burn wounds. The findings indicated that bleeding during graft skin harvest could be limited to 50 ml by intradermal injection of epinephrine (1:2,000,000), high-concentration epinephrine-soaked gauze compression (1:20,000), and temporary porcine skin coverage. Use of a scalp graft also carried a low risk of complications, with only four (2.7%) major complications including three cases (2.0%) of visible alopecia and one case (0.7%) of hair transplantation. There were no hypertrophic scars, even in the patient who had the largest number (11) of repeat harvests.

Adolescent↗

The interaction of glomerular mesangial cells and epithelial cells.

The interaction of cells within the glomerulus plays an important role in the development and progression of glomerular disease. To investigate the interaction of glomerular mesangial cells (GMC) and epithelial cells (GEC), and mediator(s) of this interaction, we investigated the effect of Adriamycin (doxorubicin hydrochloride)-induced (ADR) rat GMC-conditioned medium (GMC-CM) on the incorporation of 35S, 3H-leucine, and 3H-thymidine in normal rat GEC, as well as 3H-thymidine uptake by normal rat GMC in response to ADR-rat GEC-CM. In addition, changes in the responsiveness to interleukin-6 (IL-6) and the products of IL-6 were assessed in ADR-rat GMC. The results showed that: (1) GMC-CM of ADR-rat with heavy proteinuria stimulated GEC proliferation and the synthesis of sulfated compounds and protein, while the GEC-CM of ADR-rat from the same nephrotic period increased GMC proliferation; (2) the ADR-rat GMC had altered responsiveness to IL-6 and its products. The stimulation index results demonstrated the interaction of GMC and GEC in the ADR-induced rat model, and that this interaction related closely to the degree of proteinuria and was mediated by soluble products of the damaged glomerular cell.

Animals↗