Search PubMed⌕ Search

Biomedical subjects

J Kim

Publications and source records attributed to J Kim.

At least 955 records · Page 53Linked to original sources
PubMed1994

Phosphorylation reverses the membrane association of peptides that correspond to the basic domains of MARCKS and neuromodulin.

Several groups have observed that phosphorylation causes the MARCKS (Myristoylated Alanine-Rich C Kinase Substrate) protein to move off cell membranes and phospholipid vesicles. Our working hypothesis is that significant membrane binding of MARCKS requires both hydrophobic insertion of the N-terminal myristate into the bilayer and electrostatic association of the single cluster of basic residues in the protein with acidic lipids and that phosphorylation reverses this electrostatic association. Membrane binding measurements with myristoylated peptides and phospholipid vesicles show this hydrophobic moiety could, at best, barely attach proteins to plasma membranes. We report here membrane binding measurements with basic peptides that correspond to the phosphorylation domains of MARCKS and neuromodulin. Binding of these peptides increases sigmoidally with the percent acidic lipid in the phospholipid vesicle and can be described by a Gouy-Chapman/mass action theory that explains how electrostatics and reduction of dimensionality produce apparent cooperativity. The electrostatic affinity of the MARCKS peptide for membranes containing 10% acidic phospholipids (10(4) M-1 = chi/[P], where chi is the mole ratio of peptide bound to the outer monolayer of the vesicles and [P] is the concentration of peptide in the aqueous phase) is the same as the hydrophobic affinity of the myristate moiety for bilayer membranes. Phosphorylation decreases the affinity of the MARCKS peptide for membranes containing 15% acidic lipid about 1000-fold and produces a rapid (t1/2 < 30 s) dissociation of the peptide from phospholipid vesicles.

Amino Acid Sequence↗
PubMed1994

Multiple sequence alignment using simulated annealing.

Multiple sequence alignment is a useful technique for studying molecular evolution and analyzing structure-sequence relationships. Dynamic programming of multiple sequence alignment has been widely used to find an optimal alignment. However, dynamic programming does not allow for certain types of gap costs, and it limits the number of sequences that can be aligned due to its high computational complexity. The focus of this paper is to use simulated annealing as the basis for developing an efficient multiple sequence alignment algorithm. An algorithm called Multiple Sequence Alignment using Simulated Annealing (MSASA) has been developed. The computational complexity of MSASA is significantly reduced by replacing the high-temperature phase of the annealing process by a fast heuristic algorithm. This heuristic algorithm facilitates in minimizing the solution set of the low-temperature phase of the annealing process. Compared to the dynamic programming approach, MSASA can (i) use natural gap costs which can generate better solution, (ii) align more sequences and (iii) take less computation time.

Algorithms↗
PubMed1994

Chlorophyll regulates accumulation of the plastid-encoded chlorophyll proteins P700 and D1 by increasing apoprotein stability.

Chlorophyll protein accumulation in barley (Hordeum vulgare L.) chloroplasts is controlled posttranscriptionally by light-induced formation of chlorophyll a. The abundance of translation initiation complexes associated with psbA, psaA, and rbcL mRNAs was measured using extension and inhibition analysis in plants grown in the dark for 4.5 d and then illuminated for up to 16 h. Light-induced accumulation of the chlorophyll proteins was not accompanied by changes in the abundance of translation initiation complexes, indicating that regulation of chlorophyll protein accumulation at this stage of development does not occur at the level of translation initiation. Translational runoff assays were performed in the presence of lincomycin, an inhibitor of translation initiation, to determine whether chlorophyll protein accumulation was regulated at the level of translation elongation. The extent of ribosome runoff of psaA and psbA mRNAs was similar in the presence or absence of chlorophyll, indicating that chlorophyll did not alter chlorophyll protein translation elongation. Polysome-associated D1 translation intermediates were radiolabeled in the presence or absence of chlorophyll, even though full-length D1 accumulated only in the presence of chlorophyll. Chlorophyll influenced the stability of D1 translation intermediates to a small extent and greatly increased D1 stability after release from ribosomes. Overall, these results demonstrate that light-induced chlorophyll biosynthesis triggers the accumulation of the chlorophyll proteins D1 and P700 in barley chloroplasts by enhancement of chlorophyll apoprotein stability.

Apoproteins↗
PubMed1994

Resetting the pitch-analysis system. 2. Role of sudden onsets and offsets in the perception of individual components in a cluster of overlapping tones.

Experiments on young adults studied the effects of suddenness of onset or offset on the discrimination of the order of pitches of individual tones in a 1-s, 4-tone cluster of overlapping pure tones. In experiment 1, the tones, all within a critical band, went on asynchronously. Each rose and decayed linearly in amplitude. Faster onsets, within the range 10 to 640 ms as measured on the first tone, increased the accuracy of the discrimination of the order of onsets, but 10-ms onsets were slightly worse than 40-ms onsets in early sessions. Experiment 2 found similar effects for the abruptness of offsets of tones in clusters whose components came on synchronously but went off asynchronously. Onset order was very much easier to detect than offset order. The auditory system may use neural onset and offset responses to reset itself and carry out new analyses at frequency-by-amplitude points of sudden amplitude change, thereby contributing to auditory scene analysis.

Acoustic Stimulation↗
PubMed1994

Cloning, sequencing, and expression of the structural genes for the cytochrome and flavoprotein subunits of p-cresol methylhydroxylase from two strains of Pseudomonas putida.

The structural genes for the flavoprotein subunit and cytochrome c subunit of p-cresol (4-methylphenol) methylhydroxylase (PCMH) from Pseudomonas putida NCIMB 9869 (National Collection of Industrial and Marine Bacteria, Aberdeen, Scotland) and P. putida NCIMB 9866 were cloned and sequenced. The genes from P.putida NCIMB 9869 were for the plasmid-encoded A form of PCMH, and the genes from P.putida NCIMB 9866 were also plasmid encoded. The nucleotide sequences of the two flavoprotein genes from P.putida NCIMB 9869 and P.putida NCIMB 9866 (pchF69A and pchF66, respectively) were the same except for 5 bases out of 1,584, and the translated amino acid sequences were identical. The nucleotide sequences of the genes for the cytochrome subunits of PCMH from the two bacteria (pchC69A and pchC66) varied by a single nucleotide in their 303-base sequences, and the translated amino acid sequences differed by a single residue at position 41 (Asp in PchC69A and Ala in PchC66). Both cytochromes had 21-residue signal sequences, as expected for periplasmic proteins, and these sequences were identical. On the other hand, no signal sequences were found for the flavoproteins.pchF69A and pchC69A were expressed, separately or together, in Escherichia coli JM109 and P.putida RA4007, with active PCMH produced in both bacteria. The E. coli-expressed flavocytochrome was purified. Our studies indicated that the E.coli-expressed subunits were identical to the subunits expressed in P.putida NCIMB 9869: molecular weights, isoelectric points, UV-visible spectra, and steady-state kinetic parameters were the same for the two sets of proteins. The subunits readily associated upon mixing two crude extracts of E.coli, one extract containing PchC69A and the other containing PchF69A. The courses of association of PchC69A and PchF69A were essentially identical for pure E. coli-expressed subunits and pure P. putida 9869-expressed subunits. E. coli-expressed PchC69A and PchF69A contained covalently bound heme and covalently bound flavin adenine dinucleotide, respectively, as the proteins expressed in nature.

Amino Acid Sequence↗
PubMed1994

Localization of the Vpx packaging signal within the C terminus of the human immunodeficiency virus type 2 Gag precursor protein.

Viral protein X (Vpx) is a human immunodeficiency virus type 2 (HIV-2) and simian immunodeficiency virus accessory protein that is packaged into virions in molar amounts equivalent to Gag proteins. To delineate the processes of virus assembly that mediate Vpx packaging, we used a recombinant vaccinia virus-T7 RNA polymerase system to facilitate Gag protein expression, particle assembly, and extracellular release. HIV genes were placed under control of the bacteriophage T7 promoter and transfected into HeLa cells expressing T7 RNA polymerase. Western immunoblot analysis detected p55gag and its cleavage products p39 and p27 in purified particles derived by expression of gag and gag-pol, respectively. In trans expression of vpx with either HIV-2 gag or gag-pol gave rise to virus-like particles that contained Vpx in amounts similar to that detected in HIV-2 virus produced from productively infected T cells. Using C-terminal deletion and truncation mutants of HIV-2 Gag, we mapped the p15 coding sequence for determinants of Vpx packaging. This analysis revealed a region (residues 439 to 497) downstream of the nucleocapsid protein (NC) required for incorporation of Vpx into virions. HIV-1/HIV-2 gag chimeras were constructed to further characterize the requirements for incorporation of Vpx into virions. Chimeric HIV-1/HIV-2 Gag particles consisting of HIV-1 p17 and p24 fused in frame at the C terminus with HIV-2 p15 effectively incorporate Vpx, while chimeric HIV-2/HIV-1 Gag particles consisting of HIV-2 p17 and p27 fused in frame at the C terminus with HIV-1 p15 do not. Expression of a 68-amino-acid sequence of HIV-2 containing residues 439 to 497 fused to the coding regions of HIV-1 p17 and p24 also produced virus-like particles capable of packaging Vpx in amounts similar to that of full-length HIV-2 Gag. Sucrose gradient analysis confirmed particle association of Vpx and Gag proteins. These results demonstrate that the HIV-2 Gag precursor (p55) regulates incorporation of Vpx into virions and indicates that the packaging signal is located within residues 439 to 497.

Amino Acid Sequence↗
PubMed1994

The enhancer-blocking suppressor of Hairy-wing zinc finger protein of Drosophila melanogaster alters DNA structure.

Insertion of the gypsy retrotransposon of Drosophila melanogaster into a gene control region can repress gene expression. The zinc finger protein (SUHW) encoded by the suppressor of Hairy-wing [su(Hw)] gene binds to gypsy and prevents gene enhancers from activating transcription. SUHW blocks an enhancer only when positioned between the enhancer and promoter. Although position dependent, SUHW enhancer blocking is distance independent. These properties indicate that SUHW does not interact with the transcription activator proteins that bind to enhancers. To explore if DNA distortions are involved in enhancer blocking, the ability of SUHW to alter DNA structure was examined in gel mobility assays. Indeed, SUHW induces an unusual change in the structure of the binding-site DNA. The change is not a directed DNA bend but correlates with loss of sequence-directed bends in the unbound DNA. The DNA distortion requires a SUHW protein domain not required for DNA binding, and mutant proteins that fail to alter DNA structure also fail to eliminate the sequence-directed bends. These results suggest that SUHW increases DNA flexibility. The DNA distortion is not sufficient to block enhancers, and therefore it is suggested that increased DNA flexibility may help SUHW interact and interfere with proteins that support long-distance enhancer-promoter interactions.

Animals↗
PubMed1994

MR imaging of prostate cancer with an endorectal surface coil technique: correlation with whole-mount specimens.

PURPOSE: To evaluate the diagnostic accuracy of magnetic resonance (MR) imaging in staging prostate cancer with an endorectal surface coil technique. MATERIALS AND METHODS: The authors prospectively evaluated MR images obtained with an endorectal surface coil from 70 consecutive patients with known prostate cancer. Gadopentetate dimeglumine was administered to 40 patients. Multiple sequences were used, including conventional and fast spin echo, with and without fat suppression. The readers were blinded to the MR findings unless bone or nodal metastasis was present. MR images were compared with whole-mount sections. RESULTS: The prospective staging accuracy for MR imaging was 51% (36 of 70 patients). Stage B disease was present in 27 patients (38%), stage C in 42 (60%), and stage D in one (1%). The retrospective staging accuracy was 67% (47 of 70 patients). Of the 42 patients with stage C disease, positive surgical margins were present in 36 (85%). Gadopentetate dimeglumine did not help detect or stage tumors. CONCLUSION: Further studies must be performed to determine the role of endorectal coil MR imaging in the staging of prostate cancer.

Aged↗
PubMed1994

Differentiation of intercalated cells in developing rat kidney: an immunohistochemical study.

Intercalated cells are present in both the collecting duct, which is derived from the ureteric bud, and the connecting tubule (CNT), which is part of the nephron and thus is developed from the metanephric blastema. However, the embryologic origin of the intercalated cells has not been established. Two populations of intercalated cells, type A and type B, exist in the CNT and the cortical collecting duct (CCD). It is uncertain, however, whether these cells represent truly distinct cell types or whether one is derived from the other. In this study we have used specific antibodies to carbonic anhydrase II (CA II), H(+)-adenosinetriphosphatase (H(+)-ATPase), and band 3 protein to identify subpopulations of intercalated cells, to determine the site and time of their appearance, and to follow their differentiation in the developing rat kidney. Prenatal kidneys from 16-, 17-, 18-, and 20-day-old fetuses, and postnatal kidneys from 0-, 3-, 7-, 14-, and 21-day-old pups were preserved for immunohistochemical studies. Immunostaining for CA II and H(+)-ATPase appeared simultaneously in a subpopulation of cells in the CNT and the medullary collecting duct (MCD) of the 18-day-old fetus, suggesting that intercalated cells differentiate from separate foci, one in the nephron and one in the collecting duct. Cells with apical and cells with basolateral labeling for H(+)-ATPase appeared in the CNT and MCD at 18 days of gestation, indicating that type A and type B cells differentiate simultaneously during renal development. Band 3 immunostaining was very weak in the fetal kidney, but a striking increase in labeling was observed in the 3-day-old kidney, suggesting that there is an activation of acid-secreting cells shortly after birth. In the fetal kidney, immunostaining for CA II and H(+)-ATPase was observed in cells throughout the MCD and on the papillary surface. After birth, immunostaining gradually disappeared from both the papillary surface and the terminal inner MCD, and cells with basolateral labeling for H(+)-ATPase gradually disappeared from the outer MCD. The results of this study suggest that type A and type B intercalated cells represent distinct cell types that derive from undifferentiated cells at two separate foci, one in the nephron and one in the collecting duct. Our results also suggest that entire populations of intercalated cells are eliminated from the collecting duct during normal renal development.

Aging↗
PubMed1994

Alpha 4-integrins mediate antigen-induced late bronchial responses and prolonged airway hyperresponsiveness in sheep.

Eosinophils and T lymphocytes are thought to be involved in allergic airway inflammation. Both cells express the alpha 4 beta 1-integrin, very late antigen-4 (VLA-4, CD49d/CD29); alpha 4-integrins can promote cellular adhesion and activation. Therefore, we examined the in vivo effects of a blocking anti-alpha 4 monoclonal antibody, HP 1/2, on antigen-induced early and late bronchial responses, airway hyperresponsiveness, inflammatory cell influx, and peripheral leukocyte counts in allergic sheep. Sheep blood lymphocytes, monocytes, and eosinophils expressed alpha 4 and bound HP 1/2. In control sheep, Ascaris antigen challenge produced early and late increases in specific lung resistance of 380 +/- 42% and 175 +/- 16% over baseline immediately and 7 h after challenge, respectively, as well as airway hyperresponsiveness continuing for 14 d after antigen challenge. Treatment with HP 1/2 (1 mg/kg, i.v.) 30 min before antigen challenge did not affect the early increase in specific lung resistance but inhibited the late-phase increase at 5-8 h by 75% (P < 0.05) and inhibited the post-antigen-induced airway hyperresponsiveness at 1, 2, 7, and 14 d (P < 0.05, for each time). Intravenous HP 1/2 given 2 h after antigen challenge likewise blocked late-phase airway changes and postchallenge airway hyperresponsiveness. Airway administration of HP 1/2 (16-mg dose) was also effective in blocking these antigen-induced changes. Response to HP 1/2 was specific since an isotypic monoclonal antibody, 1E6, was ineffective by intravenous and aerosol administration. Inhibition of leukocyte recruitment did not totally account for the activity of anti-alpha 4 antibody since HP 1/2 neither diminished the eosinopenia or lymphopenia that followed antigen challenge nor consistently altered the composition of leukocytes recovered by bronchoalveolar lavage. Because airway administration of HP 1/2 was also active, HP 1/2 may have inhibited cell activation. Reduction of platelet-activating factor-induced eosinophil peroxidase release from HP 1/2-treated eosinophils supports such a mechanism. These findings indicate a role for alpha 4-integrins in processes that lead to airway late phase responses and persisting airway hyperresponsiveness after antigen challenge.

Aerosols↗
PubMed1994

Effect of Ca2+ on morphogenesis of HVJ (Sendai virus) virion in LLC-MK2 cells: suppression of viral production.

Production of HVJ (Sendai virus) virion is suppressed in the infected LLC-MK2 cells in the low Ca2+ condition. To clarify this phenomenon, the effect of Ca2+ on HVJ morphogenesis was examined under normal and low Ca2+ conditions. The HVJ production was observed in standard medium (containing 1.8 mM Ca2+) within 24 hr after infection and increased with the culture time. Indirect immunofluorescent staining with an anti-HVJ envelope antibody showed that the viral glycoproteins, HANA (HN) and F proteins, were synthesized within 10 hr after infection, diffused to the cell surface, and then many patches strongly stained with this antibody were formed on the cell surface. In contrast, when the infected cells were cultured in low Ca2+ medium, viral glycoproteins were synthesized as in the case of standard medium. However, the glycoproteins were almost all accumulated in the Golgi apparatus, suggesting that expression of viral glycoproteins at the cell surface was suppressed, and the patches were hardly seen on the cell surface. When the low Ca2+ medium was replaced by standard medium, the accumulated viral glycoproteins were rapidly diffused and expressed on the cell surface, HVJ production was restored and patches formed on the cell surface. By confocal laser scanning microscopy, the results were seen more clearly; Ca2+ is necessary for the expression of viral glycoproteins at the cell surface. By electron microscopy under normal conditions, groups of viral budding sites were seen collected in restricted areas, corresponding to the patches observed on immunofluorescent staining, but the budding sites were not seen on the cells cultured in low Ca2+ medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Further analysis of the effect of Ca2+ on morphogenesis of HVJ (Sendai virus) in LLC-MK2 cells: effects on phosphorylated M protein associated with viral morphogenesis.

Previously we have reported that Ca2+ concentration in culture medium affects the morphogenesis of HVJ from infected cells. We further investigated the effects of Ca2+ on HVJ morphogenesis in LLC-MK2 cells, focusing on M (matrix) protein which plays an important role in the assembly of viral components. On culture in standard medium (containing 1.8 mM Ca2+), most of the M protein accumulated on the apical surface in patches. On double immunofluorescent staining with anti-M protein monoclonal antibody and polyclonal antibody against HVJ glycoproteins, M protein and viral glycoproteins were observed in the same patches at the cell surface. By contrast, in low Ca2+ medium, M protein was evenly dispersed over the cell surface, especially in the regions of cell-cell contact, and was not seen in patches. Immunoblot analysis for M protein in the infected cells showed that the total amount of M protein was almost the same irrespective of Ca2+ concentration in the culture medium. But, there was a distinct difference in the forms of M protein found in the cells: most of the M protein in the cells cultured in standard medium was of the non-phosphorylated form, whereas the phosphorylated form of M protein was distinctly increased in the cells cultured in low Ca2+ medium. These results suggest that Ca(2+)-deficiency in the medium blocks dephosphorylation of phosphorylated M protein necessary for patch formation, resulting in disturbance of viral morphogenesis.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1994

[Status of parasitic infection diagnosed by surgical biopsy in Kwangju and Chollanam-do].

In order to know the species and frequency of human parasitic infection diagnosed by biopsy, 149 cases (0.18%) of parasitic infection were reviewed, which were selected from 80,947 biopsied materials submitted for routine histopathological examination during a period of 10 years from 1980 to 1989 at Department of Pathology, Chonnam National University Hospital. They consisted of 112 cases of cysticercosis, 17 paragonimiasis, 7 clonorchiasis, 4 amebiasis, 1 sparganosis, 1 enterobiasis, 1 anisakiasis, and 1 fascioliasis respectively. Based on morphological preservation of cysticercus, they could be divided into mild (20.2%), moderate (40.4%), and severe (39.4%) degeneration. Except 2 cases biopsied at the lungs, 15 cases of ectopic paragonimiasis were located at abdominal cavity (8 cases) and central nervous system (7 cases). One case of intrahepatic fascioliasis was observed. This is the 13th human fascioliasis reported in Korea. From the above results, the frequency of parasitic infections found in biopsied specimens was on the decrease as the year passed by, but biopsy is very useful diagnostic method on tissue parasites such as cysticercosis and ectopic paragonimiasis.

Adolescent↗
PubMed1994

[Chronological observation on intestinal histopathology and intraepithelial lymphocytes in the intestine of rats infected with Metagonimus yokogawai].

The relationship between the intestinal histopathology and number and position of intraepithelial lymphocytes (IEL) was observed chronologically in the small intestine of rats experimentally infected with Metagonimus yokogawai. Fifteen Sprague-Dawley rats were orally infected each with 3,000 metacecariae, and 3 were kept uninfected for controls. Three rats each were sacrificed on the day 5, 10, 15, 24 and 70 post-infection (PI) and samples of the small intestine, 5 cm, 10 cm, 20 cm and 70 cm posterior to the pylorus were taken. The samples were processed routinely and stained with Giemsa. The intestinal histopathology was severe during the day 5-15 PI and characterized by villous atrophy, crypt hyperplasia, and decrease of villus/crypt height ratio. After the day 24 PI, the intestinal lesions showed some tendency of recovery. The number of IEL increased at the early stage of infection, but decreased thereafter to a lower level than that of controls, with progression of the pathological changes. Then, the IEL number began to increase again after the day 24 PI. In control rats, the great majority of the IEL were located at the basal region of the epithelium. During the early stage of infection, however, a considerable proportion of IEL was found to have moved to the intermediate or apical region of the epithelium. From the above results, it is suggested that the change of IEL number and position during the course of M. yokogawai infection should be closely related to the progression and recovery of the intestinal histopathology.

Animals↗
PubMed1994

Resetting the pitch-analysis system: 1. Effects of rise times of tones in noise backgrounds or of harmonics in a complex tone.

The question of whether sudden increases in the amplitude of pure-tone components would perceptually isolate them from a more complex spectrum was investigated in two experiments. In Experiment 1, a 3.5-sec noise was played as a masker. During the noise, two pure-tone components of different frequencies appeared in succession. Subjects were asked to judge whether the pitch sequence went up or down. The rise time of these components had only a small and inconsistent effect on discrimination. In Experiment 2, the 3.5-sec background signal was a complex tone. The amplitudes of two of its components were incremented in succession. Again, subjects judged whether the pitch pattern went up or down. This time there was a sizable, monotonic effect of the rise time of the increments, with more rapid increments leading to better discrimination. The difference between the two results is interpreted in terms of the auditory system's response to changing and unchanging signals and the role of its "sudden-change" responses in attracting perceptual processing to certain spectral regions.

Adult↗