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H Kim

Publications and source records attributed to H Kim.

At least 721 records · Page 40Linked to original sources

Regional expression and chromosomal localization of the delta opiate receptor gene.

The delta opiate receptor gene has been cloned from the mouse neuroblastoma-rat glioma hybrid cell NG108-15. The clone that we isolated is apparently identical to that reported by Evans et al. [Evans, C. J., Keith, D. E., Jr., Morrison, H., Magendzo, K. & Edwards, R. H. (1992) Science 258, 1952-1955] and essentially identical with that of Kieffer et al. [Kieffer, B. L., Befort, K., Gaveriaux-Ruff, C. & Hirth, C. G. (1992) Proc. Natl. Acad. Sci. USA 89, 12048-12052]. We have found full-length transcripts of the gene in mouse brain but in no other tissues examined. Within the brain the gene is expressed at low levels in many regions but transcripts are found in particularly large amounts in the anterior pituitary and pineal glands. Since these tissues are located outside the blood-brain barrier, opioid peptides easily can reach receptors in these areas from the blood. The gene, which is present as a single copy, has been mapped to the distal region of mouse Chromosome 4.

Amino Acid Sequence↗

Gene organization and primary structure of a ribosomal RNA gene cluster from Streptomyces griseus subsp. griseus.

The Streptomyces griseus subsp. griseus KCTC 9080 genome contains six rRNA-encoding gene (rDNA) clusters. One rDNA cluster (rrnE), contained in an 8.7-kb BamHI fragment, was cloned and sequenced. The rDNA were arranged in the order 16S-23S-5S, and separated by small intergenic spacers. No tRNA-encoding sequences were found in the spacer regions. The lengths of the mature 16S, 23S and 5S rRNAs were 1528, 3120 and 120 nucleotides (nt), respectively. The G + C content of the gene cluster was lower than that of the chromosomal DNA. In general, the primary and secondary structures of the three rRNAs showed good agreement with those from other Streptomyces species. However, in comparison with Escherichia coli, two noticeable changes (mismatches and deletions) and two large insertions were identified in the 16S and 23S rRNAs, respectively. On the other hand, regions showing considerable heterogeneity, even within the genus Streptomyces, were found in both rRNAs. Putative primers and processing signals showing high sequence similarity to those from other Streptomyces species were located in the region upstream from the 5' end of the mature 16S rRNA. A potential hairpin loop structure reminiscent of a Rho-independent terminator was located just downstream from the 5S rRNA. A considerable degree of sequence conservation and variation within rDNA gene clusters was revealed in this study, both at the infra- and suprageneric levels.

Base Sequence↗

Synthesis and degradation of cyclic ADP-ribose by NAD glycohydrolases.

Cyclic adenosine diphosphoribose (cADPR), a recently discovered metabolite of nicotinamide adenine dinucleotide (NAD), is a potent calcium-releasing agent postulated to be a new second messenger. An enzyme that catalyzes the synthesis of cADPR from NAD and the hydrolysis of cADPR to ADP-ribose (ADPR) was purified to homogeneity from canine spleen microsomes. The net conversion of NAD to ADPR categorizes this enzyme as an NAD glycohydrolase. NAD glycohydrolases are ubiquitous membrane-bound enzymes that have been known for many years but whose function has not been identified. The results presented here suggest that these enzymes may function in the regulation of calcium homeostasis by the ability to synthesize and degrade cADPR.

Adenosine Diphosphate Ribose↗

X-ray crystallographic determination of the structure of bovine lens leucine aminopeptidase complexed with amastatin: formulation of a catalytic mechanism featuring a gem-diolate transition state.

The structure of the complex of bovine lens leucine aminopeptidase (blLAP) with the slow-, tight-binding inhibitor amastatin has been determined by X-ray crystallography. X-ray diffraction data were collected at -150 degrees C from a single blLAP-amastatin crystal which under the data collection conditions was of the space group P6(3)22 with unit cell parameters a = 130.3 A and c = 121.9 A. The structure of the blLAP-amastatin complex was determined by molecular replacement, using the structure of native blLAP as the starting model. Refinement of the blLAP-amastatin model plus 132 water molecules against data from 10.0- to 2.4-A resolution resulted in a final structure with a crystallographic residual of 0.198. The binding mode of amastatin is similar to that of bestatin, the structure of whose complex with blLAP has previously been determined. Of particular note, the N-terminus-to-C-terminus orientation of the two bound inhibitors is the same. The two N-terminal residues of amastatin and bestatin occupy the same binding sites, which are most likely S1 and S'1. The slow binding of amastatin and bestatin may be partially attributable to a binding mechanism in which the two active site metals are sequentially coordinated by the P1 amino and hydroxyl groups of these inhibitors. A catalytic mechanism for blLAP is proposed based on the binding modes of amastatin and bestatin and plausible binding modes of a dipeptide substrate and its putative gem-diolate transition state which were modeled into the active site of blLAP after the binding mode of amastatin. The proposed catalytic mechanism invokes roles for the catalytic metals in binding and activating the substrate and in stabilizing the transition state. The mechanism also includes roles for Asp-255 as a general base, Arg-336 as an additional electrophilic substrate activator and transition state stabilizer, and Lys-262 as a proton shuttle.

Amino Acid Sequence↗

Position of cyclization in cyclic ADP-ribose.

Cyclic adenosine diphosphoribose (cADPR) is a putative second messenger of calcium homeostasis synthesized from NAD by cleavage of the nicotinamide-ribose bond and cyclization of the ribose to the adenine ring. In this study, the ultraviolet absorption spectra of cADPR have been studied as a function of pH and compared to other compounds containing an adenine ring with substitutions at known positions. The results support a structure for cADPR in which cyclization is to position 1 of the adenine ring, rather than to N6, as has been previously proposed.

Adenine↗

Differentiation and identification of the two catalytic metal binding sites in bovine lens leucine aminopeptidase by x-ray crystallography.

The tightly binding and readily exchanging metal binding sites in the active site of bovine lens leucine aminopeptidase (blLAP; EC 3.4.11.1) have been differentiated and identified by x-ray crystallography. In native blLAP,Zn2+ occupies both binding sites. In solution, site 1 readily exchanges Zn2+ for other divalent cations, including Mg2+. The Zn2+ in site 2 is unavailable for metal exchange under conditions which allow exchange at site 1. The Zn2+/Mg2+ metal hybrid of blLAP (Mg-blLAP) was prepared in solution and crystallized. X-ray diffraction data to 2.9-A resolution were collected at -150 degrees C from single crystals of Mg-blLAP and native blLAP. Comparisons of omit maps calculated from the Mg-blLAP data with analogous maps calculated from the native blLAP data show electron density in one of the metal binding sites in Mg-blLAP which is much weaker than the electron density in the other binding site. Since there are fewer electrons associated with Mg2+ than with Zn2+, the difference in electron density between the two metal binding sites is consistent with occupancy of the weaker electron density site by Mg2+ and identifies this metal binding site as site 1, defined as the readily exchanging site. The present identification of the metal binding sites reverses the previous presumptive assignment of the metal binding sites which was based on the structure of native blLAP [Burley, S. K., David, P. R., Sweet, R. M., Taylor, A. & Lipscomb, W. N. (1992) J. Mol. Biol. 224, 113-140]. According to the residue-numbering convention of native blLAP, the new assignment of the metal binding sites identifies the readily exchanging site 1 with Zn-488, which is within interaction distance of one side-chain carboxylate oxygen from each of Asp-255, Asp-332, and Glu-334 and the main-chain carbonyl oxygen of Asp-332. The more tightly binding site 2 is identified with Zn-489, which is within interaction distance of one side-chain carboxylate oxygen from each of Asp-255, Asp-273, and Glu-334 and the side-chain amine nitrogen of Lys-250.

Amino Acid Sequence↗

Identification of two missense mutations in a dihydrolipoamide dehydrogenase-deficient patient.

The molecular basis of dihydrolipoamide dehydrogenase (E3; dihydrolipoamide:NAD+ oxidoreductase, EC 1.8.1.4) deficiency in an E3-deficient patient was studied. Fibroblasts cultured from the patient contained only approximately 6% of the E3 activity of cells from a normal subject. Western and Northern blot analyses indicated that, compared to control cells, the patient's cells had a reduced amount of protein but normal amounts of E3 mRNA. Direct sequencing of E3 cDNA derived from the patient's RNA as well as each of the subclones of the cDNA revealed that the patient had two substitution mutations in the E3 coding region. One mutation changed a single nucleotide from A to G, resulting in substitution of Glu (GAA) for Lys-37 (AAA). The other point mutation was a nucleotide change from C to T, resulting in the substitution of Leu (CTG) for Pro-453 (CCG). These mutations appear to be significant in that they alter the active site and possibly the binding of FAD.

Amino Acid Sequence↗

Re-refinement of the X-ray crystal structure of bovine lens leucine aminopeptidase complexed with bestatin.

Bestatin, (2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-leucine, has been incorrectly modelled in the previously reported structure of the complex between bovine lens leucine aminopeptidase (blLAP) and bestatin. In the previously reported structure, the C2 of bestatin was modelled and refined in the R configuration instead of the correct S configuration. The structure of the blLAP-bestatin complex has been re-refined after remodelling bestatin in its correct stereochemistry.

Animals↗

Expression of hydrogenase in Hupc strains of Bradyrhizobium japonicum.

Plasmid-borne hup-lacZ transcriptional fusion constructs were introduced into three separate mutant strains of Bradyrhizobium japonicum which express hydrogenase constitutively (Hupc strains SR470, SR473 and JH101) in both autotrophic and heterotrophic environments. The lacZ structural gene linked directly to the regulatory region upstream of the hydrogenase structural gene encompassing -149 bases expressed beta-gal at a constant, high level, in response to various concentrations of Ni (0 microM to 1 microM). beta-Gal activity was expressed at a constant level in response to variations in concentration of O2 (0%-10%) and H2 (0%-10%) as well. The cis-acting region required to express hydrogenase constitutively is located between -149 and -98 bases. This is also the site of nickel, oxygen and hydrogen-dependent regulatory action in the wild-type strain. It is postulated that a single mutation in Hupc strains affects the trans-acting factor which would normally by responsive to Ni, O2 and H2.

Base Sequence↗

The renal hemodynamic response to endothelin in chronic cyclosporine-treated dogs.

Use of the immunosuppressive agent cyclosporine A (CyA) is limited by its associated nephrotoxicity, characterized by an increase in renal vascular resistance (RVR) and reductions in renal blood flow (RBF) and glomerular filtration rate (GFR). The vascular endothelium produces vasoactive substances including endothelium derived relaxation factor (EDRF) and endothelin (ET), which modulate vascular tone. Since CyA has been shown to damage the endothelium, we examined the renal hemodynamic response to intrarenal ET infusion (4 micrograms./kg./minute) in chronic cyclosporine-treated dogs. Prior to ET infusion, CyA-treated dogs had a lower RBF and a greater RVR than normal dogs. In normal dogs, after ET infusion RVR increased from 30.24 +/- 0.64 to 44.60 +/- 1.66 mmHg./ml./minute (p < 0.001), RBF decreased from 4.26 +/- 0.28 to 2.90 +/- 0.30 ml./min./g. (p < 0.001) and GFR decreased from 50.20 +/- 5.90 to 36.50 +/- 7.90 ml. per minute (p < 0.001). In contrast, there was no change in RBF, GFR and RVR after intrarenal ET infusion in CyA-treated dogs. Prior to ET infusion, arterial plasma ET concentration was 5.0 +/- 1.1 pg./ml. in CyA-treated dogs, similar to 7.5 +/- 1.4 pg./ml. in normal dogs, and was not significantly altered in either group after intrarenal ET infusion. We conclude that ET may not contribute to the increased RVR in chronic cyclosporine nephrotoxicity, and suggest a vascular toxicity of CyA, rendering renal vessels unresponsive to the vasoconstrictive effect of ET.

Animals↗

Structural and functional differentiation of follicular and oviductal mouse oocytes visualised with FITC-protein conjugates.

The fluorescence labelling characteristics of mouse oocytes were examined at various stages of periovulatory differentiation using FITC-protein conjugates. The zona pellucida, perivitelline space and plasma membrane underwent visible changes which were developmentally and environmentally related. Following exposure to fluorescein isothiocyanate (FITC)-casein conjugates, the zona pellucida (ZP) of germinal vesicle stage (GV) ovarian oocytes exhibited a bright, amorphous, mesh-like staining pattern (immature type). In contrast, mature polar body stage (PB) oocytes, either ovarian or oviductal, displayed faint, spotty fluorescence labelling of the ZP (mature type). The perivitelline space (PVS) of mature ovarian oocytes (12 h post-hCG) failed to label, whereas approximately 50% of oviductal oocytes showed PVS labelling. The incidence of PVS staining increased with postovulatory age, possibly as a result of the accumulation of materials secreted by the oviduct. Following in vivo or in vitro fertilisation of oocytes, a characteristic pattern of plasma membrane (PM) labelling was observed. Similar patterns of PM labelling were seen in oocytes parthenogenetically activated with ethanol or ionophore (A23187) but not in control oocytes. The pattern of PM labelling observed with FITC-protein conjugates was strikingly similar to that observed with FITC-labelled lectins, which are thought to interact with glycoconjugates released from cortical granules. Immature type of ZP staining also occurred when GV oocytes were treated with FITC alone or with a variety of FITC-protein conjugates. Thus, protein may not be required for labelling of the ZP by FITC-protein conjugates as previously thought. FITC-conjugated proteins including casein, bovine serum albumin, peroxidase and non-immune immunoglobulin G (IgG), all labelled the PM of activated oocytes; however, FITC-IgG failed to label the PVS. Results demonstrate for the first time that various components of viable mouse oocytes exhibit and undergo characteristic structural and functional changes during periovulatory differentiation as evidenced by their interaction with one or more FITC-protein conjugates and/or FITC. On the basis of these results the intrafollicular and oviductal mechanisms mediating these changes are discussed as is the possibility that the fluorescent molecule attached to conjugates may play a role in oocyte labelling.

Animals↗

History of urethral catheters and their balloons: drainage, anchorage, dilation, and hemostasis.

Male urethral catheterization, as practiced in early Greece, was made possible by a sound knowledge of anatomy. It has been preserved by written tradition and advanced by the development of new materials. Balloon dilation, conceived and practiced more than 160 years ago by urologists, has undergone a similar cycle. Some useful ideas, once conceived, have been forgotten and have surfaced again, decades, sometimes centuries, later as new inventions.

Catheterization↗

Composite lymphoma and related disorders.

Lymphomas evolve over time, usually from the small-cell to the large-cell category, and from follicular histologic characteristics to diffuse architecture. The histologic characteristics of lymphomas may be discordant in patients with multiple sites of involvement. When different types of lymphomas are encountered in a single organ or tissue, they are designated as CLs in the Working Formulation of NHL. The most common CLs consist of different subsets of follicular center cell lymphomas, usually one with low-grade follicular histologic characteristics and another with diffuse architecture and/or more aggressive cytologic features. These lymphomas now are considered to represent different phases of clonal evolution rather than representing a coincidental simultaneous occurrence of two unrelated lymphomas. Whether B-cell large-cell lymphoma coexisting with another B-cell lymphoma, namely, the nodular variant of lymphocyte-predominant Hodgkin's disease, also represents clonal evolution requires further study. Composite lymphomas consisting of a B-cell lymphoma and a T-cell lymphoma are extremely rare. The histogenetic implications for a clonal relationship between component subsets in these and other equally rare combinations remain uncertain. The CLs should continue to be recognized because (1) the component morphologic subsets may have entirely different natural histories, requiring different treatment modalities although they may be clonally related; and (2) the study of such cases may provide us with information regarding the complex interrelationship of the lymphoid system and its clonal evolution. The morphologic definition for CL of the Working Formulation should continue to be used whenever possible, with addition of appropriate immunologic and/or molecular-genetic data.

Hodgkin Disease↗

The association of education with coronary heart disease mortality in the USSR Lipid Research Clinics Study.

There is a strong inverse association between educational attainment and coronary heart disease (CHD) mortality in men in the USSR Lipid Research Clinics (LRC) Study. Less educated men were characterized by higher mean blood pressure, high density lipoprotein cholesterol (HDL-C), cigarettes smoked and by lower mean low density lipoprotein cholesterol (LDL-C) and body mass index (BMI). With respect to nutritional variables, less educated men were characterized by higher mean energy per kg body weight and alcohol intake and by lower mean intake of fat, saturated, mono- and polyunsaturated fatty acids, cholesterol, sucrose, and other sugars. The Keys score closely predicted the differences in plasma cholesterol among the educational groups in the randomly selected sample. In men without CHD at entry, the age and clinic-adjusted relative risk for CHD mortality was 2.4 for the least educated compared with the most educated group; on adjusting for systolic blood pressure, HDL-C, LDL-C, BMI, alcohol intake and number of cigarettes, the relative risk was reduced to 1.9. These data indicate that only 22% of the twofold excess of CHD mortality associated with low education was statistically attributable to the major risk factors. The failure to explain more of the education-CHD mortality gradient in the USSR LRC cohort was similar to observations from cohort studies in Great Britain and the US. Other correlates of low education must be explored to explain the association.

Body Mass Index↗

A comparison of human and bovine Escherichia coli O157:H7 isolates by toxin genotype, plasmid profile, and bacteriophage lambda-restriction fragment length polymorphism profile.

Foods of bovine origin have been linked to human disease outbreaks caused by Escherichia coli O157:H7 and may be linked to the more common sporadic cases as well. In this study, E. coli O157:H7 from the bovine reservoir (22 isolates: 12 from dairy and 10 from beef breed cows) and from human patients (50 isolates from sporadic human infections) were compared using Shiga-like toxin genotypes, plasmid profiles, and DNA restriction fragment length polymorphisms identified with a bacteriophage lambda probe (lambda-RFLP). Twenty-three lambda-RFLP profiles, 4 Shiga-like toxin genotypes, and 8 plasmid profiles were identified among the isolates tested. Together the typing methods distinguished 43 strains, of which 3 were isolated from both humans (5 isolates) and cattle (6 isolates; 5 from dairy herds). These data demonstrate the value of lambda-RFLP as a means of strain identification for E. coli O157:H7.

Animals↗

Use of interleukin-7, interleukin-2, and interferon-gamma to propagate CD4+ T cells in culture with maintained antigen specificity.

In contrast to CD8+ T cells, it has been difficult to establish consistently satisfactory conditions for the bulk culture of antitumor CD4+ T cells in either mice or humans. This difficulty is not limited to tumor antigen, since similar problems are encountered growing CD4+ T cells that recognize alloantigen, tetanus, or candida. Four basic findings are reviewed in this article, stemming from work with identical results in both human and mouse. (a) Although CD4+ T cells initially proliferate after exposure to appropriate antigen-presenting cells (APCs), such proliferation is not sustained; however, expansion of CD4+ T cells can be achieved with the addition of recombinant interleukin-2 (rIL2) or rIL-7. (b) Specific CD4+ responses to antigens are often better sustained with exogenous IL-7 than with exogenous IL-2. (c) Adding rIL-7 or rIL-2 permits sustained CD4+ T-cell proliferation, but subsequent culture restimulation is complicated by high background reactivity to APCs whether APCs are antigen pulsed or not; this problem can be overcome by addition of exogenous interferon-gamma (IFN-gamma) to the CD4+ cultures. (d) In certain instances proliferation is paradoxically impaired by reexposure to specific antigen, possibly reflecting apoptosis; this problem is also overcome by addition of rIFN-gamma to culture. We conclude that combinations of exogenous IL-7, IL-2, and IFN-gamma with APC restimulation can be used to sustain antigen-specific CD4+ T cells in culture. Using these techniques, antitumor CD4+ T cells were propagated from the peripheral blood of two tumor-bearing patients.

Animals↗