Search PubMed⌕ Search

Biomedical subjects

K Han

Publications and source records attributed to K Han.

At least 73 records · Page 4Linked to original sources
PubMed1998

Recombinant flounder growth hormone from Escherichia coli: overexpression, efficient recovery, and growth-promoting effect on juvenile flounder by oral administration.

An efficient production method for recombinant flounder growth hormone (r-fGH) from Escherichia coli was developed and the biological activity of purified r-fGH was examined using juvenile flounder. The use of bicistronic construction in the expression plasmid resulted in the production of over 40% of the E. coli cellular protein as r-fGH. The r-fGH was recovered from cell lysates following inclusion body washing, solubilization and refolding in sodium dodecylsulfate (SDS) solution, and removal of contaminated proteins with secondary butanol treatment. The SDS content in purified r-fGH solution was adjusted to appropriate levels by diafiltration. More than 47% of the r-fGH was recovered from the E. coli cell lysates and the purity of recovered r-fGH was 98%. The oral administration of purified r-fGH to juvenile flounder, once a week for 4 weeks at a dosage of 40 micrograms r-fGH g-1 fish body weight, resulted in significant increases both in weight and length. These results of overexpression, simple purification with high recovery yield and purity, and good growth-promoting activity of the r-fGH suggest that the production scheme described in this study is useful for the potential application of r-fGH in fish farming.

Administration, Oral↗
PubMed1998

A comparison of multileaf-collimator and alloy-block field shaping.

PURPOSE: The purpose of this report was to compare the dose distribution at a field edge defined with divergent alloy blocks to the distribution obtained with a multileaf collimator (MLC). The comparison is made for simple block replacement situations. METHODS AND MATERIALS: A tertiary multileaf collimator mounted on a linear accelerator operating at 6 MV was compared to divergent alloy blocks positioned at the level of the blocking tray. The leaves of the MLC were positioned to give maximum stepping (leaf displacement equals leaf width), and the blocking produced the same field shape. Three different treatment plans were compared: single field, opposed fields, and a four-field "box." Dose distributions were determined using radiographic film scanned with a laser densitometer with a 0.45-mm spot size. One experiment was repeated using radiochromic film with reduced energy dependence. Dose distributions were examined on the isocenter plane, and on planes displaced by 1.0 and 2.5 cm. The effect of daily setup variations was also studied by comparing a single fraction treatment with a fractionated treatment consisting of 15 fields slightly displaced relative to each other. The magnitude of these displacements was determined using available literature on treatment reproducibility. RESULTS: For a single field plan, maximum stepping of an MLC-defined edge produces an obvious undulating dose pattern compared to an alloy block edge. At the isocenter plane, this pattern is unchanged when parallel opposed fields are used. However, blurring occurs for both MLC and block edges when planes displaced from the isocenter are examined. The gradient for the block edge is 8%/mm for opposed fields and a plane 2.5 cm from the isocenter, compared to 15%/mm for the isocenter plane. Adding two additional fields does not change the dose pattern in the isocenter plane, but does reduce the gradient across the steepest portion of the penumbra to 8%/mm, and shifts the isodose line with the most pronounced stepping to higher values (from 50 to 80%). Introducing daily setup variations results in a reduction of the sharp dose gradient along the sides of a single field, and around the periphery of the beam at the isocenter plane of opposed fields. Smaller changes are found for edges already blurred by other factors. Radiochromic film was generally noisier than radiographic film, but comparison of the two films did not show a significant difference, indicating that the energy dependence of the radiographic film was not a problem. CONCLUSIONS: The obvious dose stepping seen on a portal image of a single field with MLC shaping is shown to be partially erased by the addition of other fields, and for planes away from the isocenter. However, the effects of daily setup variations must be included to more effectively blur dose stepping along the external envelope of a single field or near the isocenter plane of opposed fields. This result conflicts with attempts to improve immobilization.

Particle Accelerators↗
PubMed1998

Metabolism of an anionic fluorescent dye, 1-anilino-8-naphthalene sulfonate (ANS) by rat liver microsomes.

The present study was designed to examine the metabolism of 1-anilino-8-naphthalene sulfonate (ANS), an anionic compound which is transported into liver via "multispecific organic anion transporter", with rat hepatic microsomes. TLC analysis indicated that the fluorescent metabolites were not produced to a measurable extent, which made it possible to assess the ANS metabolism by measuring the fluorescence disappearance. The metabolism of ANS was remarkably inhibited by the presence of SKF-525A as well as by the substitution of O2 by CO gas. ANS metabolism by microsomes also required NADPH as a cofactor. These results indicated that the microsomal monooxygenase system might be mainly responsible for the ANS metabolism. The maximum velocity (Vmax) and Michaelis constant (K(m)) were calculated to be 4.3 +/- 0.2 nmol/min/mg protein and 42.1 +/- 2.0 microM, respectively. Assuming that 1 g of liver contains 32 mg of microsomal protein, the Vmax value was extrapolated to that per g of liver (Vmax'). The intrinsic metabolic clearance (CLint) under linear conditions calculated from this in vitro metabolic study was 3.3 ml/min/g liver, being comparable with that (3.0 ml/min/g liver) calculated by analyzing the in vivo plasma disappearance curve in a previous study. Furthermore, the effects of other organic anions on the metabolism of ANS were examined. Bromophenolblue (BPB) and rose bengal (RB) competitively inhibited the metabolism of ANS, while BSP inhibited it only slightly. The inhibition constant (Ki) of BPB (6 microM) was much smaller than that of RB (200 microM). In conclusion, the microsomal monooxygenase system plays a major role in the metabolism of ANS, and other unmetabolizable organic anions (BPB and RB) compete for this metabolism.

Anilino Naphthalenesulfonates↗
PubMed1998

Ocular absorption of Pz-peptide and its effect on the ocular and systemic pharmacokinetics of topically applied drugs in the rabbit.

PURPOSE: To determine the corneal and conjunctival penetration of 4-phenylazobenzyloxycarbonyl-L-Pro-L-Leu-Gly-L-Pro-D-Arg (Pz-peptide) and to evaluate its effect on the corneal and conjunctival penetration of hydrophilic solutes as well as on the ocular and systemic absorption of topically applied atenolol and propranolol in the rabbit. The hydrophilic solutes were mannitol, fluorescein, FITC-dextran 4,000, and FITC-dextran 10,000. METHODS: Drug penetration across the rabbit cornea and conjunctiva was evaluated using the modified Ussing chamber. Ocular and systemic absorption of topically applied atenolol and propranolol was evaluated by analyzing the drug concentration in various anterior segment tissues at 45 min and in the blood over 240 min, respectively, following topical instillation of 25 microl of 20 mM atenolol or propranolol solution to the rabbit eye. RESULTS: The conjunctiva was 29 times more permeable than the cornea to 3 mM Pz-peptide. Conjunctival Pz-peptide transport was 1.7 times more extensive in the mucosal-to-serosal than in the opposite direction, whereas corneal Pz-peptide transport showed no directionality. The apparent permeability coefficient of Pz-peptide across the cornea and the conjunctiva increased over the 1-5 mM range, suggesting that Pz-peptide enhanced its own transport across both epithelial tissues. The cornea appeared to be more sensitive than the conjunctiva to the penetration enhancement effect of Pz-peptide. Thus, whereas Pz-peptide elevated the corneal transport of mannitol, fluorescein, and FD4 by 50%, 57%, and 106%, respectively, it did not affect the conjunctival transport of mannitol and fluorescein, while enhancing FD4 transport by only 46%. Moreover, while Pz-peptide enhanced the ocular absorption of topically applied hydrophilic atenolol, it did not affect the ocular absorption of lipophilic propranolol. Interestingly, Pz-peptide did not affect the systemic absorption of either beta adrenergic antagonist. CONCLUSIONS: Pz-peptide appears to facilitate its own penetration across the cornea and the conjunctiva. Pz-peptide appears to increase the ocular absorption of topically applied hydrophilic but not lipophilic drugs, while not affecting the systemic absorption of either type of drugs.

Actins↗
PubMed1998

Acute lymphoblastic leukemia with maturation--a new entity with clinical significance.

The diagnosis of 'ALL with maturation' (ALLm) is proposed. One hundred and one patients with untreated ALL were entered into this study. The diagnosis of ALLm was made when more than 20% of all nucleated elements in the bone marrow showed maturation beyond prolymphocytes by light microscopic examination. The mature-appearing leukemic cells showed the same immunophenotype to remaining lymphoblasts. The number of ALLm cases was 19 (18.8%). The mean age at presentation of ALLm was 29 +/- 18, older than that of 18 +/- 16 of the remaining typical ALL (ALLt) (P = 0.015). Remission was induced with daunorubicin, vincristine, prednisone and L-asparaginase. Only two of 19 ALLm patients achieved CR after 4 weeks induction chemotherapy. In contrast, 57 of 82 (69.5%) ALLt patients achieved CR after the same induction chemotherapy. There was no significant difference in immunophenotype of ALLm compared with ALLt. Labeling index of DNA topoisomerase IIalpha (TopoLI) was studied by immunohistochemistry. Initial TopoLI of ALLm (221 +/- 147) was much lower than that of ALLt (609 +/- 262, P = 0.005). Furthermore, the remaining leukemic cells after chemotherapy were not labeled with anti-DNA topoisomerase IIalpha. The P53 protein was expressed in nine of 18 ALLm cases (50.0%) and P-glycoprotein was not expressed in ALLm cases. Twelve of 19 ALLm cases were studied for carrying bcr/abl fusion by karyotyping and/or fluorescent in situ hybridization. Only two cases revealed bcr/abl fusion. In conclusion, ALLm is a separate entity of ALL which has a very poor clinical course and is independent of other prognostic factors. The morphologically mature leukemic cells are in resting GO phase.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

Cloning of the Alcaligenes latus polyhydroxyalkanoate biosynthesis genes and use of these genes for enhanced production of Poly(3-hydroxybutyrate) in Escherichia coli.

Polyhydroxyalkanoates (PHAs) are microbial polyesters that can be used as completely biodegradable polymers, but the high production cost prevents their use in a wide range of applications. Recombinant Escherichia coli strains harboring the Ralstonia eutropha PHA biosynthesis genes have been reported to have several advantages as PHA producers compared with wild-type PHA-producing bacteria. However, the PHA productivity (amount of PHA produced per unit volume per unit time) obtained with these recombinant E. coli strains has been lower than that obtained with the wild-type bacterium Alcaligenes latus. To endow the potentially superior PHA biosynthetic machinery to E. coli, we cloned the PHA biosynthesis genes from A. latus. The three PHA biosynthesis genes formed an operon with the order PHA synthase, beta-ketothiolase, and reductase genes and were constitutively expressed from the natural promoter in E. coli. Recombinant E. coli strains harboring the A. latus PHA biosynthesis genes accumulated poly(3-hydroxybutyrate) (PHB), a model PHA product, more efficiently than those harboring the R. eutropha genes. With a pH-stat fed-batch culture of recombinant E. coli harboring a stable plasmid containing the A. latus PHA biosynthesis genes, final cell and PHB concentrations of 194.1 and 141.6 g/liter, respectively, were obtained, resulting in a high productivity of 4.63 g of PHB/liter/h. This improvement should allow recombinant E. coli to be used for the production of PHB with a high level of economic competitiveness.

Acyltransferases↗
PubMed1998

Reconstruction of three-dimensional digital teeth.

OBJECTIVE: Dental anatomy is one of the most important basic courses in the education of dentistry. The deep understanding of both the external and internal morphologic characteristics is very important to the teaching, research and clinical practice of dentistry. METHODS: In the present study, 32 permanent teeth from the skull specimen of a young man were individually embedded in black fluid resin, each in special containers. They were ground by a numerically-controlled grinding machine at intervals of 0.2 mm per layer. A distinct outline of the tooth could be seen on every section. Black and white photographs were taken and scanned into a computer by a film-scanner to obtain sequences of two-dimensional images of the tooth sections. After pattern recognition, all of the images of each tooth were piled up at intervals of 0.2 mm by the technique of computerized reconstruction. RESULTS: The three-dimensional stereo tooth models were built up from two-dimensional data. On the basis of those models, computer graphic techniques were used to highlight, smooth, and shade the teeth. These digital teeth and dentitions could function as a series of computerized teaching models. Their high resolution and accuracy could meet the basic demands of dentistry. CONCLUSIONS: The present study developed new techniques of model preparation, image input, and 3-D reconstruction. These digital teeth and dentitions provide an important foundation for the application of computer imaging, CAD/CAM, computer-assisted instruction, and virtual reality in dentistry.

Anatomy, Cross-Sectional↗
PubMed1998

[Development of emission models for volatile organic compounds from indoor materials].

Volatile organic compounds (VOCs) emitted from indoor materials was a major cause of indoor air pollution. The characteristics of VOCs emission was an important part of research programs on indoor air quality. The technology of test chambers with exactly controllable conditions has been successfully used in studies of VOCs emissions. The technology could be used to model the chamber VOCs concentration level vs time profile C(t), which could in turn be used to estimate the sample emission rate vs time profile R(t). The emission models of VOCs from indoor materials were presented in this review. The principal of emission process, parameters and the applications of emission models were introduced. The application of diffusion model, dilution model and vapor pressure (VP) model were limited due to the existence of sink effect. Sink model is the most promising model at present.

Air Pollution, Indoor↗
PubMed1998

[Three dimensional reconstruction of human permanent teeth].

OBJECTIVE: Dental anatomy is one of the most important basic courses in the education of stomatology. The deep understanding of morphologic characters is closely instructive to the clinic of endodontics, oral surgery, orthodontics and prosthodontics. METHODS: In present study, 32 permanent teeth from a skull specimen were inputted into computer after the processes of burying, grinding, photographing, scanning and recognizing. By the techniques of reconstruction, the 3D stereo tooth models were retrieved from 2D digital data. RESULTS: On the basis of those 3D data, computer graphics techniques were used to realize the lighted, smoothed and shaded teeth. CONCLUSION: Such a series of data teeth might be applicated as a corner stone of the computer aided instruction (CAI) in stomatology.

Computer-Assisted Instruction↗
PubMed1997

Promoter region of the Drosophila melanogaster norpA gene.

We have sequenced the 5'-flanking region of the Drosophila melanogaster norpA gene and characterized its promoter. The potential promoter region, which was deduced from the determination of the transcription start point (tsp), lacks a distinct TATA box sequence. Deletion analysis of the promoter region suggests that the minimal promoter necessary for efficient expression of the gene is located between -138 (PstI) and +278 relative to the tsp. Within this minimal promoter region, at least two downstream regulatory elements responsible for the stimulation of gene expression seem to exist in the DNA fragments between +44 and +121 and between +214 and +278. Among these, the DNA fragment between +44 and +121 affects promoter activity more dramatically (about 6-7 fold). This DNA fragment contains the consensus promoter element previously reported to be important for photoreceptor cell-specific expression, and this promoter element seems to be working in the norpA gene expression.

Animals↗
PubMed1997

Advanced glycation endproducts stimulate mitogen-activated protein kinase and proliferation in rabbit vascular smooth muscle cells.

Advanced glycation end products of bovine serum albumin (AGEs-BSA) exhibited biphasic effects on the proliferation of cultured rabbit vascular smooth muscle cells (VSMCs) in terms of [3H]thymidine incorporation and cell number count; a stimulatory effect was observed at 1-10 micrograms/ml and an inhibitory effect at more than 20 micrograms/ml, while it inhibited [3H]thymidine incorporation even at 1-10 micrograms/ml in cultured bovine vascular endothelial cells (VECs). Transient activation of p42 mitogen-activated protein kinase (MAPK) with a peak at around 5 min and a subsequent sustained phase was induced by AGEs-BSA in VSMCs, but not in VECs. The dependence of MAPK activation on AGEs-BSA dose was correlated with that of VSMCs proliferation.

Animals↗
PubMed1997

Bombesin stimulates the motility of human prostate-carcinoma cells through tyrosine phosphorylation of focal adhesion kinase and of integrin-associated proteins.

Bombesin-like peptides, including the mammalian homologue gastrin-releasing peptides, are highly expressed and secreted by neuroendocrine cells in prostate carcinoma (PCa) tissues and are likely to be related to the progression of this disease. In the present study, we show that bombesin enhances the migration of androgen-independent PCa cells (PC-3) in vitro, while not affecting their adhesion to extracellular matrix proteins. The bombesin-increased motility of PC-3 cells occurs through its receptor, and, as shown with inhibitors, it likely requires activation of both protein tyrosine kinases (PTKs) and protein kinases C (PKCs). Because the focal adhesion kinase pp125FAK plays a key role in adhesion/motility and is highly expressed in advanced PCa, we examined whether in PC-3 cells bombesin signal transduction triggers the tyrosine phosphorylation of this PTK and of associated integrins and signaling proteins likely to be present in focal adhesion plaques. pp125FAK tyrosine phosphorylation was stimulated by bombesin and mimicked by PKC activation with the tumor-promotor phorbol 12-myristate-13-acetate (PMA). Moreover, this effect of bombesin on pp125FAK tyrosine phosphorylation requires the presence of both active PKC and cytoskeleton integrity since this signal was abolished by down-regulating PKCs induced by prolonged PMA treatment or by PKC inhibition with GF 109203X, as well as by disruption of the cytoskeleton with cytochalasin D. We also show that bombesin increases the tyrosine phosphorylation of a 95-kDa protein (pp95) which was co-immunoprecipitated with the alpha v and beta (3 and 5) subunits, forming integrin receptors with alpha v in PC-3 cells. The protein pp95 is distinct from the endogenously tyrosine-phosphorylated beta3 subunit. In addition, upon bombesin treatment, the beta1, beta3 and beta5 integrin subunits co-immunoprecipitated with pp125FAK and major phosphotyrosine (pY)-containing proteins of 125 and 68-70 kDa, likely corresponding to pp125FAK and paxillin. Together our data suggest that, in addition to PKC activation, tyrosine phosphorylation of pp125FAK and integrin-associated proteins may play an important role in bombesin signaling, triggering the processes of PCa cell motility and invasion.

Bombesin↗
PubMed1997

Analysis of the structure and expression fo the TFIIB gene in Drosophila melanogaster.

We have isolated and characterized a genomic clone encoding the Drosophila melanogaster transcription factor IIB (TFIIB). The coding region of the TFIIB gene is interrupted by three short introns. The 5'-flanking region of the gene lacks the typical TATA box sequence like those of other known genes encoding the general transcription factors. In addition, the 5'-flanking region of the gene contains several common DNA sequences present in Drosophila TBP and TFIIS genes, suggesting the common regulation mechanism of gene expression. RNA blot analysis revealed that the gene expresses 1.6 kb, 1.3 kb and 1.2 kb mRNAs throughout development and in adults. Deletion analysis of the promoter region shows that the minimal promoter necessary for efficient expression is located between -698 (PstI) and +60 relative to the transcription start point. Within this minimal promoter region, the upstream regulatory element responsible for the stimulation of gene expression may exist in the DNA fragment between -698 (PsfI) and -351 (StuI).

Amino Acid Sequence↗