Biomedical subjects

M K Kim

Publications and source records attributed to M K Kim.

At least 109 records · Page 6Linked to original sources

PubMed1999

In vivo transcription factor recruitment during thyroid hormone receptor-mediated activation.

Thyroid hormone receptor (TR) can act as both a transcriptional activator and a silencer. Optimal activation by TR requires synergism with activator(s) bound to the promoter (promoter proximal activator). It is thought that liganded TR either helps to recruit preinitiation complexes (PIC) to the promoter or activates the PIC already recruited. However, the studies analyzing the TR action on the PIC formation were done in vitro and, therefore, it is not clear how relevant they are to the in vivo TR action. For example, in vivo, the TR can act from distances equal to or greater than a kilobase from the promoter, but such distant effect is not reproducible in vitro. In this study, we used the PIN*POINT (ProteIN POsition Identification with Nuclease Tail) assay to define the molecular mechanism of TR action on transcription from the thymidine kinase promoter in the cellular context. We demonstrate that the recruitment of promoter-proximal activator Sp1, and the components of the basal transcription factors such as TBP, TFIIB, and Cdk7, is enhanced with thyroid hormone activation. Our results suggest that DNA forms a loop with TR-mediated activation to accommodate interactions between the liganded TR complex and the complex formed on the promoter. We also show that Sp1 bound to the promoter is essential for the DNA looping and recruitment of basal transcription factors such as TFIIB and Cdk7 but not for recruitment of TBP. On the basis of these findings, we present a model that illustrates the molecular mechanism of TR-mediated activation in vivo.

Base Sequence↗

PubMed1999

Cloning and expression of the gene encoding phospholipase A1 from Serratia sp. MK1 in Escherichia coli.

The gene encoding extracellular phospholipase A1 of Serratia sp. MK1 was cloned from a genomic DNA library. Formation of transparent halos on the PCY agar plates was used to identify E. coli carrying the phospholipase A1 gene. A 4.2 kb EcoRI fragment was isolated and sequenced. From nucleotide sequences and expression of various plasmids, two open reading frames (plaA and plaS) involved in efficient expression of phospholipase A1 in natural and recombinant host were identified. Extracellular phospholipase A1 activity was identified as the gene product of plaA encoding 321 amino acids with a predicted MW of 33,400. Analysis of the amino acid sequence revealed significant homology (around 70%) to phospholipase A1 of Serratia liquefaciens and Yersinia enterocolitica. The sequence, -Gly-X1-Ser-X2-Gly-, known as a lipase-specific consensus sequence was also found in the bacterial phospholipase A1. PlaS encoding a protein of 224 amino acids showed no enzymatic activity, but might be necessary for the efficient expression of phospholipase A1 in E. coli. To further improve the production of phospholipase A1 as a soluble and active form in E. coli, the effect of some parameters was examined. Surprisingly, a higher yield of soluble and active phospholipase A1 could be obtained under the combined conditions of a lower temperature, an enriched medium, and a lower-strength promoter.

Amino Acid Sequence↗

PubMed1999

Acyl-CoA: cholesterol acyltransferase inhibitory activity of ginseng sapogenins, produced from the ginseng saponins.

Ginseng sapogenins were produced from ginseng saponins, isolated from Korean ginseng roots. Ginseng saponins very mildly inhibited acyl-CoA:cholesterol acyltransferase (ACAT) in vitro, however, the sapogenins showed strong inhibitory activity on microsomal ACAT. Therefore, the sapogenins will be one of key ingredients of ginseng affected a lowering of the serum total cholesterol level.

Animals↗

PubMed1999

Design and development of new collimator cones for fractionated stereotactic radiation therapy in Samsung Medical Center.

PURPOSE: In stereotactic radiotherapy using X-Knife system, the commercially supplied collimator cone system had a few mechanical limitations. The authors have developed new collimator cones to overcome these limitations and named them "SMC type" collimator cones. METHODS: We made use of cadmium-free cerrobend alloy within the stainless steel cylinder housing. We made nine cones of relatively larger sizes (3.0 cm to 7.0 cm in diameter) and of shorter length with bigger clearance from the isocenter than the commercial cones. The cone housing and the collimator cones were designed to insert into the wedge mount of the gantry head to enable double-exposure linac-gram taking. RESULTS: The mechanical accuracy of pointing to the isocenter was tested by ball test and cone rotation test, and the dosimetric measurements were performed, all of which were with satisfactory results. A new innovative quality assurance procedure using linac-grams on the patients at the actual treatment setup was attempted after taking 10 sets of AP and lateral linac-grams and the overall mechanical isocenter accuracy was excellent (average error = 0.4 +/- 0.2 mm). CONCLUSIONS: We have developed the SMC type collimator cone system mainly for fractionated stereotactic radiation therapy use with our innovative ideas. The new cones' mechanical accuracy and physical properties were satisfactory for clinical use, and the verification of the isocenter accuracy on the actual treatment setup has become possible.

Alloys↗

PubMed1999

Higher intracellular levels of uridinemonophosphate under nitrogen-limited conditions enhance metabolic flux of curdlan synthesis in Agrobacterium species.

Changes of intracellular nucleotide levels and their stimulatory effects on curdlan synthesis in Agrobacterium species were investigated under different culture conditions. Under nitrogen-limited conditions where curdlan synthesis was stimulated, intracellular levels of UMP were as high as 87 and those of AMP were 78 nmol/mg of cellular protein, while those under nitrogen-sufficient conditions were lower than 45 nmol/mg-protein. The levels of other nucleotides such as UDP, UTP, UDP-glucose, ADP, ATP, and ADP-glucose were lower than 30 nmol/mg-protein under both nitrogen-limited and sufficient conditions. The time profiles of curdlan synthesis and cellular nucleotide levels showed that curdlan synthesis had a positive relationship with intracellular levels of UMP and AMP. After the ammonium concentration in the medium fell below 0.1 g/L, intracellular levels of UMP and AMP increased, followed by curdlan synthesis. However, no significant changes in the specific activities of UMP kinase, UDP kinase, and UDP-glucose pyrophosphorylase were observed during cultivation. In vitro enzyme reactions for the synthesis of UDP-glucose, which serve as a precursor for curdlan synthesis, demonstrated that the synthesis of UDP-glucose increased with the increase of UMP concentration. In contrast, AMP had no effect on UDP-glucose synthesis at all. Addition of UMP in the medium increased the curdlan synthesis, whereas curdlan synthesis was inhibited in the presence of AMP. From these results, we concluded that only the higher intracellular UMP levels caused by nitrogen limitation in the medium enhance the metabolic flux of curdlan synthesis by promoting cellular UDP-glucose synthesis.

Adenosine Monophosphate↗

PubMed1999

The pharmacokinetic characteristics of glycolated humanized anti-Tac Fabs are determined by their isoelectric points.

To evaluate a method for preventing the nephrotoxicity caused by the high renal accumulation of radiolabeled or toxin-conjugated small immunoproteins used for cancer therapy, we conjugated humanized anti-Tac Fab fragments with various numbers of glycolate molecules [glycolated Fab fragments (glyco-Fabs)] and separated the conjugates by means of ion-exchange columns into three fractions, depending on their isoelectric points (pIs). We evaluated the biodistribution, pharmacokinetics, and catabolism in normal nude mice of nonglycolated Fab (pI > or = 9.3) and three different preparations of glyco-Fab, including strongly anionic glyco-Fab (sa-glyco-Fab: pI < or = 4.5), mildly anionic glyco-Fab (pI = 4.5-7), and mildly cationic glyco-Fab (pI = 7-9.3). In addition, the biodistributions of 125I-labeled sa-glyco-Fab and 131I-labeled nonglycolated Fab were evaluated in normal nude mice coinjected with 50 mg of L-lysine and/or 1 microg of furosemide and in a control group without coinjection. We then evaluated the serial biodistribution of 125I-labeled sa-glyco-Fab (4 microCi/1 microg) and 131I-labeled nonglycolated Fab (5 microCi/1 microg) in Tac antigen-positive (ATAC4) and -negative (A431) tumor-bearing nude mice with s.c. tumor xenografts derived from Tac antigen-positive ATAC4 cells and receptor-negative A431 cells. These animals were coinjected with 30 mg of lysine i.v. and 30 mg of lysine i.p. 15 min after the radiolabeled Fab injection. To evaluate the biodistribution data and study scintigraphic imaging, we performed serial scintigraphy on normal and tumor-bearing mice with all four 131I-labeled preparations. 125I-labeled mildly cationic glyco-Fab and 131I-labeled nonglycolated Fab had similar distributions, except in the kidney. However, both 125I-labeled anionic glyco-Fab preparations showed significantly different distributions from both cationic Fabs in the blood, liver, lung, and spleen. Renal accumulation of all four radiolabeled Fab preparations increased significantly as the pI increased (P < 0.01). In addition, the intact fraction of Fab excreted into urine increased as pI decreased. Therefore, the glomerular filtration depended on whether the charge on the Fab was positive or negative. The proportion of Fab reabsorbed by the proximal tubules increased as pI increased. 125I-labeled sa-glyco-Fab and 125I-labeled mildly anionic glyco-Fab showed a similar distribution in the blood and all organs except the kidney. Lysine led to an additional blocking effect on proximal tubular uptake of both sa-glyco-Fab and nonglycolated Fab. Addition of furosemide yielded only a small effect when used with lysine. With lysine, the sa-glyco-Fab:nonglycolated Fab estimated integral radioactivity ratios were 4.7 and 0.7 in the ATAC4 tumor and in the kidney, respectively. The use of anionic fragments, which may be used in conjunction with lysine, represents a promising approach that may help decrease the renal toxicity of other small fragments, the molecular weights of which range from Mr 40,000 to 70,000, and, thereby, allow higher doses of radiation to the tumor.

Animals↗

PubMed1999

Disassembly of focal adhesions during apoptosis of endothelial cell line ECV304 infected with Orientia tsutsugamushi.

Many bacterial pathogens induce apoptosis in their host cells. We observed the cellular effect of ECV304 cells infected with Orientia tsutsugamushi. The infected cells became rounded and floated in culture supernatant. These floating cells as well as adherent cells exhibited typical features of apoptosis, such as DNA fragmentation and TUNEL staining. As many cells detached from growth substrate, we examined the focal adhesion using the immunofluorescence assay method and observed decreased focal adhesions in heavily infected cells. As endothelial cells could undergo apoptosis by the loss of focal adhesions, this change of focal adhesions may account for the Orientia-induced apoptosis.

Apoptosis↗

PubMed1999

Central neck dissection for the treatment of recurrent thyroglossal duct cysts in childhood.

OBJECTIVE: The recurrence rate of thyroglossal duct cysts removed by the classic Sistrunk procedure exceeds 4%, even in skilled hands. Simple reexcision fails in 33% of these patients. Recent pathology literature suggests that the tracts of thyroglossal duct cysts may arborize, arguing for a wide-field approach to recurrent lesions. We describe the anatomic rationale and technique of an en bloc central neck dissection in children, on the basis of cadaver dissections and histopathologic review of recurrent thyroglossal duct cyst specimens. METHODS: We reviewed the medical records of all the children undergoing surgery for thyroglossal duct cysts and fistulas during the years 1990 to 1998 by the senior author. En bloc central neck dissections were performed on several cadaver specimens to further delineate the anatomic rationale for this procedure. RESULTS: We have performed an en bloc central neck dissection in 7 children, 5 with recurrent or multiply recurrent thyroglossal duct cysts. None has had a recurrence after follow-up of 9 months to 6 years. All have acceptable functional and cosmetic results. CONCLUSION: An en bloc central neck dissection is a logical and effective surgical technique for the removal of recurrent or multiply recurrent thyroglossal duct cysts.

Adolescent↗

PubMed1999

A randomized clinical trial of dermal anesthesia by iontophoresis for peripheral intravenous catheter placement in children.

STUDY OBJECTIVE: To assess the efficacy of dermal anesthesia by lidocaine iontophoresis in children undergoing peripheral intravenous (PIV) catheter placement in the emergency department. METHODS: A double-blind, randomized, clinical trial was conducted at a tertiary children's hospital ED. Alert children 7 years or older requiring nonemergency PIV were eligible. Patients in the lidocaine group received 1 mL of 2% lidocaine with 1:100,000 epinephrine over a potential PIV site by iontophoresis. The control group received 1 mL of.9% saline solution with 1:100,000 epinephrine. After PIV placement, patients ranked the procedural pain using a visual analog scale. Complications were noted by visual inspection or telephone follow-up. RESULTS: During a 6-month period, 22 patients were assigned to the lidocaine group and 25 to the control group. There was no significant difference in age, sex, or ethnic background between the 2 study groups, and mean application time was 12.0 minutes. The median pain score was.5 in the lidocaine group compared with 4 in the control group (P =.0002; 95% confidence interval [CI] 1 to 5). No significant immediate or delayed complications were observed. CONCLUSION: Lidocaine iontophoresis provides effective dermal anesthesia for children older than 7 years of undergoing nonemergency PIV placement in the ED.

Anesthesia, Local↗

PubMed1999

Structural requirements of Syk kinase for Fcgamma receptor-mediated phagocytosis.

The tyrosine kinase Syk plays a critical role in the phagocytic pathway mediated by Fcgamma receptors (FcgammaR). In transfected COS1 cells co-expression of Syk enhances FcgammaR mediated phagocytosis. The other member of the Syk kinase family, the highly homologous tyrosine kinase Zap70, also plays a role in signaling by immunoglobulin gene family receptors, but does not increase the phagocytic efficiency of FcgammaRs. The homologous tandem SH2 and kinase domains of Syk and Zap70 are separated by a nonhomologous region referred to as the unique domain. Zap70's inability to enhance phagocytosis was not due to unique domain tyrosine 292, previously implicated in negative regulation of Zap70 function. We determined the regions of Syk important for its interaction with the phagocytic pathway. An intact kinase domain was required for Syk's effect on phagocytosis. Furthermore, the Syk variant SykB, lacking 23 amino acids in the unique region, signaled for phagocytosis as efficiently as did Syk. We then constructed exchange chimeras between Syk and Zap70 and determined the contributions of the SH2, unique and kinase domains to phagocytic signaling. Our data suggest that the Syk kinase domain, which has high intrinsic kinase activity, is important for facilitating phagocytic signaling by FcgammaRI and FcgammaRIIIA.

Animals↗

PubMed1999

Evaluation of condylar neck fracture plating techniques.

The purpose of this study was to compare the biomechanical stability of four different plating techniques used to fix condylar neck fractures and to decide which fixation systems are strong enough to withstand the functional load. Ten recently acquired formalin-fixed cadaver mandibles were used for this study. Each of the four sets of osteotomized condylar processes was fixed by one of four different fixation systems. The mandibles were then held in an angle vice so that the mandibles were oriented to simulate actual masticatory force loading on the temporomandibular joint and were loaded with an Instron loading machine. Data demonstrated that a two-miniplate system applied to the anterior and posterior regions of the condylar neck was more stable than single-plate repairs using either mini-dynamic compression plates or 2.4 mm plates. The two-miniplate-fixation technique is indicated in cases of condylar neck fracture to achieve early mobility of the jaw and stability of the fracture site.

Biocompatible Materials↗

PubMed1999

Role of actin filaments in the hatching process of mouse blastocyst.

Hatching has been suggested to occur as a result of protease-mediated lysis and the blastocoele tension. However, even if rupturing is initiated at multiple sites, interestingly only a single site is used for escape. This implies that there are several mechanisms involved in hatching. In this study, the involvement of actin filaments in mouse embryo hatching was examined. We treated mouse embryos with cytochalasin B for 12 h or 24 h at the morula, middle blastocyst, expanded blastocyst, lobe-formed blastocyst and hatching blastocyst stages, and measured the amount and distribution of actin filaments using a confocal microscope. At morula, middle blastocyst, lobe-formed blastocyst and hatching blastocyst stages embryonic development was completely arrested by cytochalasin B. However, when transferred to cytochalasin-B-free medium, the embryos resumed development and escaped the zona pellucida. In the expanded blastocysts development was almost completely inhibited by cytochalasin B, but rupturing occurred in some embryos. However, development stopped completely at the ruptured stage. Distribution of actin filaments was prominent at rupturing and hatching sites regardless of cytochalasin B treatment. The amount of actin filaments was prominent at hatching embryos compared with other developmental stages of embryos. These actin filaments were distributed intensively between the trophectodermal cells, and formed locomotion patterns. Taken together, these results suggest that not only tension and lytic enzymes are required to rupture, but the activity of actin filaments may have a crucial role in the process of hatching.

Actin Cytoskeleton↗

PubMed1999

Common head and neck manifestations of AIDS.

Head and neck manifestations of acquired immunodeficiency syndrome (AIDS) can involve the skin, ear, upper aerodigestive tract, and neck. Several head and neck manifestations of AIDS may be the only initial sign of this disease process and therefore primary-care physicians and otolaryngologists must be able to recognize and understand the management of these lesions. Cystic enlargement of the parotid gland and Kaposi's sarcoma are increasingly being encountered in the head and neck exam of HIV-infected patients. An example of each of these disease processes is presented with full discussion about the various treatment methods.

Adult↗

PubMed1999

Breaking the bad news of cancer: the patient's perspective.

OBJECTIVE: Head and neck cancers present a special challenge to the patient and the physician because they affect the quintessential aspects of living such as breathing, eating, and speaking. Numerous articles have described how the physician should perform the difficult task of conveying bad news, but only a small number of articles specifically assess the patients' perceptions when being told of a serious diagnosis. The purpose of this survey was to evaluate the thoughts and concerns of patients receiving diagnoses of head and neck cancer. STUDY DESIGN: Questionnaire survey. METHODS: A 41-item questionnaire was sent to head and neck cancer patients who have been treated for and remain free of disease for at least 2 years. RESULTS: All of the respondents felt that their diagnosis was adequately explained to them and that no further explanations were necessary. Eighty-one percent of the respondents did not wish to have anyone else present at the time of diagnosis. Additionally, 75% of the respondents did not want the physician to touch their hands or hug them when given the bad news. Only 63% of the respondents had further discussions with family, friends, or other physicians after being told of their diagnosis. CONCLUSION: When patients are told of the diagnosis of head and neck cancer, they want their physician to be truthful, caring, and compassionate. The patients want their diagnosis in simple and direct terms without the incorporation of complex medical terminology. The results of this survey can provide insightful information to physicians when they are undertaking the difficult task of conveying bad news to their patients.

Communication↗

PubMed1999

Positional preference of proline in alpha-helices.

Conformational free energy calculations have been carried out for proline-containing alanine-based pentadecapeptides with the sequence Ac-(Ala)n-Pro-(Ala)m-NHMe, where n + m = 14, to figure out the positional preference of proline in alpha-helices. The relative free energy of each peptide was calculated by subtracting the free energy of the extended conformation from that of the alpha-helical one, which is used here as a measure of preference. The highest propensity is found for the peptide with proline at the N-terminus (i.e., Ncap + 1 position), and the next propensities are found at Ncap, N' (Ncap - 1), and C' (Ccap + 1) positions. These computed results are reasonably consistent with the positional propensities estimated from X-ray structures of proteins. The breaking in hydrogen bonds around proline is found to play a role in destabilizing alpha-helical conformations, which, however, provides the favored hydration of the corresponding N-H and C=O groups. The highest preference of proline at the beginning of alpha-helix appears to be due to the favored electrostatic and nonbonded energies between two residues preceding proline and the intrinsic stability of alpha-helical conformation of the proline residue itself as well as no disturbance in hydrogen bonds of alpha-helix by proline. The average free energy change for the substitution of Ala by Pro in a alpha-helix is computed to be 4.6 kcal/mol, which is in good agreement with the experimental value of approximately 4 kcal/mol estimated for an oligopeptide dimer and proteins of barnase and T4 lysozyme.

Peptides↗

PubMed1999

Apoptosis of endothelial cell line ECV304 persistently infected with Orientia tsutsugamushi.

Endothelial cells are major targets of Orientia tsutsugamushi. To examine the consequences of the infection of endothelial cells with O. tsutsugamushi, we used human endothelial cell line ECV304. Persistent infection was established and infected cultures could be maintained for over seven months without the addition of normal cells. The heavily infected cells became round and floated in the culture medium, harboring large numbers of organisms inside them. Some of the infected ECV304 cells showed features of apoptotic cells, as determined by the terminal deoxytransferase-mediated dUTP nick end-labeling reaction and DNA fragmentation. We also found that O. tsutsugamushi increased transcription of the mRNAs of proinflammatory cytokines such as IL-6 and IL-8. These results show the first evidence of in vitro-persistent infection by O. tsutsugamushi, which may be related to in vivo persistence reported previously.

Animals↗

PubMed1999

hnRNP U inhibits carboxy-terminal domain phosphorylation by TFIIH and represses RNA polymerase II elongation.

This study describes a potential new function of hnRNP U as an RNA polymerase (Pol II) elongation inhibitor. We demonstrated that a subfraction of human hnRNP U is associated with the Pol II holoenzyme in vivo and as such recruited to the promoter as part of the preinitiation complex. hnRNP U, however, appears to dissociate from the Pol II complex at the early stage of transcription and is therefore absent from the elongating Pol II complex. When tested in the human immunodeficiency virus type 1 transcription system, hnRNP U inhibits elongation rather than initiation of transcription by Pol II. This inhibition requires the carboxy-terminal domain (CTD) of Pol II. We showed that hnRNP U can bind TFIIH in vivo under certain conditions and inhibit TFIIH-mediated CTD phosphorylation in vitro. We find that the middle domain of hnRNP U is sufficient to mediate its Pol II association and its inhibition of TFIIH-mediated phosphorylation and Pol II elongation. The abilities of hnRNP U to inhibit TFIIH-mediated CTD phosphorylation and its Pol II association are necessary for hnRNP U to mediate the repression of Pol II elongation. Based on these observations, we suggest that a subfraction of hnRNP U, as a component of the Pol II holoenzyme, may downregulate TFIIH-mediated CTD phosphorylation in the basal transcription machinery and repress Pol II elongation. With such functions, hnRNP U might provide one of the mechanisms by which the CTD is maintained in an unphosphorylated state in the Pol II holoenzyme.

Heterogeneous-Nuclear Ribonucleoprotein U↗

PubMed1999

Discordant evolution of asymptomatic proteinuria in identical twins.

We describe a pair of 17-year-old identical twin brothers with asymptomatic proteinuria, one of whom showed focal segmental glomerulosclerosis (FSGS) while the other showed immunoglobulin M (IgM) nephropathy. For each twin, audiological examination was normal. There was no family history of renal failure, deafness, or hematuria. HLA typing revealed an identical phenotype consisting of A25, A33, B44, B54, Cw1, Cw7, DR7 and DRB1. There is still controversy about whether minimal change disease, IgM nephropathy, and FSGS are discrete entities or different aspects of the same disease. The coexistence of IgM nephropathy and FSGS in identical twins suggests that the same genetic factors may be involved in the development of both diseases. However, although the brothers are identical twins, they had different eating habits and body weight. The twin who preferred to eat a protein-rich diet and who was heavier developed early proteinuria and manifested FSGS on renal biopsy. The discordant evolution of asymptomatic proteinuria in identical twins may provide a clue for the existence of environmental factors on the progression from IgM nephropathy to FSGS. Therefore, this report provides indirect support for the hypothesis that IgM nephropathy and FSGS represent different aspects in the spectrum of a single disease.

Adolescent↗