Biomedical subjects

M Kim

Publications and source records attributed to M Kim.

At least 235 records · Page 13Linked to original sources

PubMed1998

The effect of N-alkyloxycarbonyl group on the anticonvulsant activities of N-alkyloxycarbonyl-alpha-amino-N-methylsuccinimides.

In connection with the development of new anticonvulsant agents with a broad spectrum, we found that N-Cbz-alpha-amino-N-alkylsuccinimides showed significant anticonvulsant activities, and the pharmacological activities of these compounds were dependent on their stereochemistry and N-substituted alkyl group. These results prompted us to define the effects of other functional group on the anticonvulsant activities of these compounds. Therefore a series of N-alkoxycarbonyl-alpha-amino-N-methylsuccinimide were prepared from N-Cbz-aspartic acid and were evaluated with their anticonvulsant activities against the MES and PTZ tests, in order to define the effect of N-substituted alkoxy carbonyl group with the anticonvulsant activities. From these studies, it was found that all the tested N-alkoxycarbonyl-alpha-amino-N-methylsuccinimides exhibited significant anticonvulsant activities in the PTZ test and were not active in the MES test. The most active compound in the PTZ test was (S) N-ethoxycarbonyl-alpha-amino-N-methyl-succinimide. We found that the pharmacological activities in the PTZ test were dependent on their N-alkoxycarbonyl groups. They follow as such; The order of anticonvulsant activities for (R) series as evaluated by ED50 was N-phenoxycarbonyl = N-4-nitrobenzyloxycarbonyl > N-ethoxycarbonyl > N-allyloxycarbonyl > N-tert. butoxycarbonyl compound; For the (S) series N-ethoxycarbonyl > N-phenoxycarbonyl > N-allyloxycarbonyl compound. From the above results, it was conceivable that N-substituted alkoxycarbonyl group had certain effects on the anticonvulsant activities of N-alkoxycarbonyl-alpha-amino-N-methylsuccinimides.

Animals↗

PubMed1998

The effect of N-alkyloxycarbonyl group on the anticonvulsant activities of N-alkyloxycarbonyl-alpha-aminoglutarimides.

In connection with the development of new anticonvulsant agents with a broad spectrum, we reported that N-Cbz-alpha-aminoglutarimides, combining common structures of other anticonvulsants such as N-CO-C-N and cyclic imides in a single molecule, showed significant anticonvulsant activities in the MES (maximal electroshock seizure) and PTZ (pentylenetetrazole induced seizure) tests. In these studies, a series of (R) and (S) N-alkyloxycarbonyl-alpha-aminoglutarimides 7a-7e and 8a-8e, which were substituted with various alkyloxycarbonyl group instead of Cbz group, were prepared from the corresponding (R) and (S) N-Cbz-glutamic acid 3 and 4, and were evaluated with their anticonvulsant activities against the MES and PTZ tests, including neurotoxicity, in order to define the effect of N-alkyloxycarbonyl group on the anticonvulsant activities of N-alkyloxycarbonyl-alpha-aminoglutarimides. Among them, (S) N-4-nitrobenzyloxycarbonyl-alpha-amino-N-methylglutarimide 8e was the most active in MES (ED50 = 35.6 mg/kg, PI = 2.7) and PTZ tests (ED50 = 15.6, PI = 6.1). Interestingly, (R) and (S) N-4-nitrobenzyloxycarbonyl-alpha-amino-N-methylglutarimide 7e and 8e and (R) N-phenoxycarbonyl-alpha-amino-N-methylglutrimide 7d showed significant anticonvulsant activities in both the MES and PTZ tests and other compounds showed anticonvulsant activities in only the PTZ test. In addition, it was found that their anticonvulsant activities were dependent on their stereochemistries and N-substituted alkyloxycarbonyl groups.

Animals↗

PubMed1998

Comparison of egg-yolk protein hydrolysate and soyabean protein hydrolysate in terms of nitrogen utilization.

Egg-yolk protein hydrolysate (YPp) is an alternative protein source in formulas for infants with intolerance to cow's milk or soyabean protein, or for patients with intestinal disorders. However, the nutritional value of YPp has never been investigated. YPp was prepared by enzymic hydrolysis of delipidated yolk protein, which led to an average peptide length of 2.6 residues. Three experiments were performed. In Expt 1, we compared the intestinal absorption rate of YPp and soyabean protein hydrolysate (SPp) in rats. YPp and SPp solutions were injected into the duodenum of anaesthetized rats and blood samples were taken from the portal vein at 7, 15, 30, 60, and 120 min. A higher amino acid concentration in the serum of the YPp group demonstrated that YPp was absorbed faster than SPp. In Expt 2, the effects of dietary YPp and SPp on body-weight gain, protein efficiency ratio (PER) and feed efficiency ratio (FER) were determined. At the end of the experiment, body weight had increased in both groups, while PER and FER were significantly higher in rats fed on YPp. In Expt 3, to investigate the effects of dietary YPp and SPp on N metabolism, we determined the biological value and net protein utilization. Yolk protein was the reference protein. Biological value and net protein utilization values were very similar between animals fed on yolk protein and YPp diets, and significantly higher than in rats fed on the SPp diet. The present findings demonstrate that there is no adverse effect of hydrolysis of yolk protein on N utilization, and that the nutritive value of YPp is similar to that of yolk protein and superior to that of SPp.

Animals↗

PubMed1997

Promoting apoptosis: a novel activity associated with the cyclin-dependent kinase inhibitor p27.

p27Kip1, a cyclin-dependent kinase inhibitor, is recognized as a negative regulator of the cell cycle. In this paper, we report that overexpression of p27Kip1 triggers apoptosis in several different human cancer cell lines. Using a recombinant adenoviral vector that expresses p27Kip1 (Adp27), we found that overexpression of p27Kip1 in MDA-MB-231 breast cancer cells induces apoptosis that was seen by a number of different techniques, including flow cytometry and in situ terminal deoxynucleotidyl transferase-mediated nick end labeling, flow cytometric assay for sub-G1 population, and 4',6-diamindino-2-phenylindole staining. Cleavage of poly(ADP-ribose) polymerase and degradation of cyclin B1, events that are known to be associated with apoptosis, were also observed following overexpression of p27Kip1. This is the first report indicating a role for p27Kip1 in induction of apoptosis.

Adenoviridae↗

PubMed1997

Simultaneous determination of catechins in human saliva by high-performance liquid chromatography.

Green tea extracts have been suggested to possess a preventive effect against dental caries. A quantitative method for their anticariogenic substances, catechins, was developed to evaluate their concentrations in human saliva after mouthrinsing with green tea extract. Salivary catechins were extracted to the organic phase after forming a complex with diphenylborate and an ion-pair with tetra-n-butylammonium, and then back-extracted to the acidic aqueous phase. The extract was analyzed by high-performance liquid chromatography using diode array detection at absorption wavelengths ranging from 269 to 278 nm. In reversed-phase chromatography by a gradient elution, eight catechins originating from green tea and an internal standard were separated in 15 min without interfering peaks. All the catechins were simultaneously and selectively determined in the concentration range 0.05-25.0 microg/ml. In replicate spiking experiments with standards, the mean recovery ranged between 86 and 99%, and both intra- and inter-assay C.V.s were within 2.3%. When mouthrinsing with an aqueous solution of green tea extract (5.0 mg/ml) containing eight catechins, the quantitative results revealed that each catechin was retained at microg/ml levels in saliva for up to 60 min.

Catechin↗

PubMed1997

[Results of a standardized treatment concept in primary hyperparathyroidism].

BACKGROUND AND OBJECTIVE: Primary hyperparathyroidism (PHPT) is being diagnosed ever more frequently. After diabetes mellitus, ovarian dysfunction and metabolic disorders it is now among the most frequent metabolic disease. Its surgical treatment has become standardized with increasing experience. A prospective study was undertaken to clarify to what extent our own standardized diagnostic and therapeutic measures conform to current standards in literature. PATIENTS AND METHODS: During 10 years (1. 1. 1987-31. 12. 1996) 478 patients with PHPT were treated surgically (solitary adenoma: 317 (76.5%), multiple lobe hyperplasias 66 (15.9%), double adenoma 28 (6.8%), and carcinoma 4 (0.8%). PHPT was part of the MEN (multiple endocrine neoplasia) syndrome in 20 patients. Typical symptoms of PHPT had been present in 400 patients (83.7%), while 78 (16.3%) had no manifest symptoms. Ultrasound examination of the soft tissues was the only preoperative test of localization. More extensive preoperative diagnosis was practised only before reoperations. All patients had bilateral exploration of the neck to try and demonstrate all four epithelial bodies. RESULTS: A primary operation normalized the calcium level in 405 of 414 patients (97.8%). In 64 previously unsuccessfully operated patients who were reoperated for persisting HPT the elevated calcium levels could be normalized in 90.6%. Hypercalcaemia was finally abolished in all 478 patients. Average follow-up duration for 319 patients was more than 4.8 (0.5-8.5) years. The perioperative mortality rate was 0.8 (4 of 478), with 41 deaths during the later follow-up period. Permanent recurrent laryngeal nerve palsy occurred in 6 patients (1.8%). Long-term calcium substitution was necessary postoperatively in four patients (1.2%). CONCLUSION: Our findings support a liberal policy towards indication for bilateral surgical exploration of the neck in PHPT.

Adenoma↗

PubMed1997

Fluid shear stress activation of focal adhesion kinase. Linking to mitogen-activated protein kinases.

Shear stress, the tangential component of hemodynamic forces, activates the extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) signal transduction pathways in cultured vascular endothelial cells to induce the transcriptional activation of many immediate early genes. It appears that integrins, protein-tyrosine kinases, and the structural integrity of actin are important factors involved in these shear stress-induced responses. The underlying molecular events were investigated by the application of a shear stress of 12 dyn/cm2 on bovine aortic endothelial cells (BAEC). We found that such a shear stress increased the tyrosine phosphorylation and the kinase activity of focal adhesion kinase (FAK) and its association with growth factor receptor binding protein 2 (Grb2) in a rapid and transient manner, suggesting that FAK may be linked to these mitogen-activated protein kinase signaling pathways through a Grb2. Son of sevenless (Sos) complex. FAK(F397Y), which encodes a dominant negative mutant of FAK, attenuated the shear stress-induced kinase activity of Myc epitope-tagged ERK2 and hemagglutinin epitope-tagged JNK1. DeltamSos1, encoding a dominant negative mutant of Sos in which the guanine nucleotide exchange domain has been deleted, also attenuated shear stress activation of Myc-ERK2 and hemagglutinin-JNK1. Pretreating the confluent BAEC monolayers with a blocking type anti-vitronectin receptor monoclonal antibody had similar inhibitory effects in these shear stress-activated ERKs and JNKs. Confocal microscopic observation further demonstrated that FAK tended to cluster with vitronectin receptor near the abluminal side of the sheared BAEC. These results demonstrate that FAK signaling is critical in the shear stress-induced dual activation of ERK and JNK.

Actins↗

PubMed1997

Molecular assembly of the extracellular domain of P2X2, an ATP-gated ion channel.

We have produced the putative extracellular domain (ECD) of the ATP-gated ion channel, P2X2, in a bacterial expression system. The hexahistidine-tagged protein was purified by immobilized metal affinity chromatography and refolded by sulfitolysis and dialysis. We demonstrate that P2X2-ECD forms a stable tetramer in solution by gel filtration chromatography, dynamic light scattering and analytical sedimentation centrifugation. [alpha-32P]ATP has been covalently cross-linked by UV irradiation to the P2X2-ECD and this binding is specific and competable by antagonists suramin and cibacron blue. These results indicate that the binding affinity among P2X2-ECD subunits is appreciably stronger than 3.4 microM (0.1 mg/ml), implying that the extracellular domain of P2X2 is primarly responsible for tetramerization of whole P2X2 and thus probably plays a role in determining homo- and heteromerization specificity of P2X channel subunits.

Adenosine Triphosphate↗

PubMed1997

Alternative splicing in the pore-forming region of shaker potassium channels.

We have cloned cDNAs for the shaker potassium channel gene from the spiny lobster Panulirus interruptus. As previously found in Drosophila, there is alternative splicing at the 5' and 3' ends of the coding region. However, in Panulirus shaker, alternative splicing also occurs within the pore-forming region of the protein. Three different splice variants were found within the P region, two of which bestow unique electrophysiological characteristics to channel function. Pore I and pore II variants differ in voltage dependence for activation, kinetics of inactivation, current rectification, and drug resistance. The pore 0 variant lacks a P region exon and does not produce a functional channel. This is the first example of alternative splicing within the pore-forming region of a voltage-dependent ion channel. We used a recently identified potassium channel blocker, kappa-conotoxin PVIIA, to study the physiological role of the two pore forms. The toxin selectively blocked one pore form, whereas the other form, heteromers between the two pore forms, and Panulirus shal were not blocked. When it was tested in the Panulirus stomatogastric ganglion, the toxin produced no effects on transient K+ currents or synaptic transmission between neurons.

Amino Acid Sequence↗

PubMed1997

Synthesis of a novel sialic acid derivative (sialylphospholipid) as an antirotaviral agent.

A novel sialylphospholipid (SPL) was synthesized from N-acetylneuraminic acid (NeuAc) and phosphatidylcholine (PC) by a chemical and enzymatic method and evaluated as an inhibitor of rotavirus. PC and 1,8-octanediol were conjugated by transesterification reaction of Streptomyces phospholipase D (PLD) under a water-chloroform biphasic system to afford phosphatidyloctanol, which was condensed with a protected 2-chloro-2-deoxyneuraminic acid derivative by using silver trifluoromethanesulfonate as an activator in chloroform and converted, after deprotection, to SPL. Rhesus monkey kidney cells (MA-104) were incubated with simian (SA-11 strain) and human (MO strain) rotaviruses in the presence of SPL, and the cells infected were detected indirectly with anti-rotavirus antibody. SPL showed dose dependent inhibition against both virus strains. The concentrations required for 50% inhibition (IC50) against SA-11 and MO were 4.35 and 16.1 microM, respectively, corresponding to 10(3)- and 10(4)-fold increases in inhibition as compared to monomeric NeuAc.

Animals↗

PubMed1997

Additive Effects on the Microstructure Evolution in Hexadecyltrimethylammonium Bromide Solution and Its Rheological Properties

The effects of various additives on the microstructure evolution in hexadecyltrimethylammonium bromide micellar solution and its rheological properties are investigated. The additives considered in the present study are primary, secondary, and tertiary heptanols and sodium salicylate (NaSal). The microstructure is developed via the two-step shape transitions in the micellar solution; first, the initial spherical micelles undergo shape transition to rod-like or disc-like micelles as the micelles tend to be packed compactly with increase in the surfactant concentration; then further increment in the surfactant concentration makes the anisotropic rod-like micelles overlap each other. Solutions in these states exhibit typical non-Newtonian behaviors such as shear thinning at high shear rates. In the present study, the additive effects on the microstructure evolution are investigated by employing various techniques including a phase modulated flow birefringence, a dynamic light scattering and a dynamic oscillatory rheometry. The results show that addition of the solubilized additives enhances the microstructure transitions, which are affected by the additive concentration and its chemical structure. Presence of the cosurfactants such as heptanols with hydrophobic alkyl chains reduces the repulsion by forming surfactant-alcohol mixed micelles. The primary heptanol can penetrate easily into the micelles and aligned parallel to the surfactant molecules compared with the secondary and tertiary heptanols. Thus, the primary heptanol enhances the two-step shape evolutions more effectively than the other types. In the presence of NaSal, on the other hand, the micellar solution exhibits the viscoelastic properties and the yield stress owing to the formation of networked or worm-like micelles. Copyright 1997 Academic Press. Copyright 1997Academic Press

Journal Article↗

PubMed1997

Identification of YH439 and its metabolites in rat urine by gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry.

YH439 is a potential drug candidate for the treatment of various hepatic disorders. YH439 and its three metabolites have been identified in rat urine by liquid chromatography-mass spectrometry (LC-MS) and by gas chromatography (GC)-MS. Identification of YH439 and its metabolites was established by comparing their GC retention times and mass spectra with those of the synthesized authentic standards. Both electron impact- and positive chemical ionization MS have been evaluated. The metabolism study was performed in the rat using oral administration of the drug. A major metabolite (YH438) was identified as the N-dealkylation product of YH439. Other identified metabolites were caused by the loss of the methyl thiazolyl amine group (metabolite II) from YH439, the isopropyl hydrogen malonate group (metabolite IV) and the decarboxylated product (metabolite III) of metabolite II.

Administration, Oral↗

PubMed1997

The subunit structure of cytochrome-c oxidase from tuna heart and liver.

Cytochrome-c oxidase was isolated from tuna liver and heart, and the subunit composition was analysed by SDS/PAGE by two separation systems. Two additional subunits of the enzyme complex were immunoprecipitated from solubilized mitochondria with an antibody against bovine subunit IV. The N-terminal and internal amino acid sequences of all nuclear-coded subunits were determined after blotting onto poly(vinylidene difluoride) membranes or by tryptic hydrolysis of gel bands and HPLC separation of peptides, respectively. 13 subunits were identified with isoforms for subunits Va, VIc, VIIb and VIII. The isoforms for subunits Va and VIIb are found in liver and heart, isoforms for subunit VIc only in heart, and isoforms for subunit VIII only in liver. Isoforms for subunits Va, VIc and VIIb have not been described in other species. The postulated mechanism of thermogenesis in mammals, based on decreased H+/e- stoichiometry at high ATP/ADP ratios due to binding of ATP to the heart-type subunit VIa [Frank, V. & Kadenbach, B. (1996) FEBS Lett. 382, 121-124], appears not to occur in tuna, because no isoforms of subunit VIa were found.

Amino Acid Sequence↗

PubMed1997

mRNAs for microtubule proteins are specifically colocalized during the sequential formation of basal body, flagella, and cytoskeletal microtubules in the differentiation of Naegleria gruberi.

We have examined the distribution of four mRNAs-alpha-tubulin, beta-tubulin, flagellar calmodulin, and Class I mRNA-during differentiation of Naegleria gruberi amebas into flagellates by in situ hybridization. Three of the four mRNAs-alpha-tubulin, beta-tubulin, and Class I mRNA-began to be colocalized at the periphery of the cells as soon as transcription of the respective genes was activated and before any microtubular structures were observable. At 70 min after the initiation of differentiation, these mRNAs were relocalized to the base of the growing flagella, adjacent to the basal bodies and microtubule organizing center for the cytoskeletal microtubules. Within an additional 15 min, the mRNAs were translocated to the posterior of the flagellated cells, and by the end of differentiation (120 min), very low levels of the mRNAs were observed. Cytochalasin D inhibited stage-specific localization of the mRNAs, demonstrating that RNA localization was actin dependent. Since cytochalasin D also blocked differentiation, this raises the possibility that actin-dependent RNA movement is an essential process for differentiation.

Actin Cytoskeleton↗

PubMed1997

A recombinant adenovirus expressing p27Kip1 induces cell cycle arrest and loss of cyclin-Cdk activity in human breast cancer cells.

In order to elucidate the biochemical mechanisms by which the universal cyclin kinase inhibitor p27Kip1 regulates cell cycle progression in human breast cancer cells, a recombinant adenovirus expressing human p27 was constructed (Adp27). Upon infection of human breast cancer cells MDA-MB-231 and MCF-7 with Adp27, a high level of p27 expression was observed, and this resulted in a marked decrease in the proportion of cells in S-phase. In multiple cell lines, comparison of the cytotoxicity of Adp27 with another adenovirus vector expressing the related universal cyclin kinase inhibitor WAF1/Cip1 (AdWAF1), showed Adp27 to be markedly more (up to 56-fold) toxic than AdWAF1. DNA histograms showed Adp27 to cause a G1/S arrest at lower viral doses than AdWAF1. Analysis of cyclin dependent kinase activity following Adp27 infections showed decreased Cdk2 and cyclin B1-Cdc2 activity at lower viral doses when compared with AdWAF1. Adp27 is therefore potentially useful for studies of growth regulation and for gene therapy when growth inhibition is desired.

Adenoviridae↗

PubMed1997

Occurence of a sialylglycopeptide and free sialylglycans in hen's egg yolk.

Free sialylglycans (FSGs) and a sialylglycopeptide (SGP) as components of hen's egg yolk were found and their chemical structures were determined. SGP and FSGs were isolated from fresh egg yolk by treatment with phenol, gel filtration and successive chromatographies on columns of anion- and cation-exchangers. They were localized in the yolk plasma. The glycan moiety of SGP, which was liberated by PNGase digestion, was studied for the chemical structure by HPLC mapping with p-aminobenzoic ethylester-derivatization, sugar composition analysis, 1H nuclear magnetic resonance and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and the glycomoiety was found to be an N-linked disialyl-biantennary glycan. The amino acid sequence of the peptide moiety of SGP was determined to consist of Lys-Val-Ala-Asn-Lys-Thr, the Asn of which is modified with the disialylglycan moiety. FSGs were determined to be two free disialyl-biantennary glycans whose reducing end was either Man beta1-4GlcNAc (FSG-I) or Man beta1-4GlcNAc beta1-4GlcNAc (FSG-II). Since the molar value of SGP present in one egg yolk (2.8 micromol) is comparable to those of well-known major yolk proteins, low density lipoprotein, lipovitellins and phosvitin, it can be considered that SGP is one of the major components in hen's egg yolk.

Amino Acids↗

PubMed1997

Induction of apoptotic cell death in a neuroblastoma cell line by dibucaine.

Dibucaine, a local anesthetic known to interact with cell membranes, induced apoptosis in SK-N-MC human neuroblastoma cells in a dose-dependent manner. Apoptosis was demonstrated by direct visualization of morphological nuclear changes using a DAPI staining technique and confirmed by the production of characteristic ladder patterns of DNA fragmentation on gel electrophoresis. At concentrations which induced apoptosis, dibucaine significantly altered membrane fluidity, indicating that fluidity may be a major target for the cytotoxic action of dibucaine. Also, dibucaine increased intracellular calcium levels more effectively in calcium-containing Krebs-Ringer buffer than in calcium-free Krebs-Ringer buffer. Removal of extracellular calcium or addition of antioxidants or protein synthesis inhibitor effectively blocked dibucaine-induced apoptosis. These results suggest that membrane damage, intracellular calcium levels, and oxygen free radicals may be involved in the apoptosis induced by dibucaine.

Anesthetics, Local↗

PubMed1997

A novel gene therapy strategy for elimination of prostate carcinoma cells from human bone marrow.

We report a novel means to purge bone marrow of a specific subset of prostate carcinoma cells based on transductional and genetic selectivity. Using both adenovirus-polylysine-DNA complexes and E1A/B-deleted replication-deficient adenoviruses, we have demonstrated a transductional preference of these vectors for the prostate carcinoma cell lines DU 145, LNCaP, and PC-3 over primary human bone marrow cells and the leukemia cell line KG-1. We have also shown a genetic selectivity of an anti-erbB-2 intracellular single-chain antibody (sFv) encoding adenovirus, Ad21, for the erbB-2-positive prostate carcinoma cell lines DU 145 and LNCaP. Delivery of Ad21 resulted in cytotoxicity to the DU 145 and LNCaP, but not PC-3, cell lines and reduced the clonogenic capacity of DU 145 cells cultured alone or mixed with various ratios of irradiated human bone marrow. Finally, quantitative, competitive reverse transcription polymerase chain reaction (QC-RT-PCR) analysis demonstrated that Ad21 could effectively reduce DU 145 and erbB-2-positive primary prostate tumor contamination in bone marrow cultures. Delivery of Ad21 had no effect on the ability of progenitor cells to form colonies. These results suggest that an anti-erbB-2 sFv-encoding adenoviral vector is efficacious for removal of erbB-2-positive prostate carcinoma cells from human bone marrow, and demonstrates a novel method for ex vivo genetic purge of malignant cells from bone marrow for autologous bone marrow transplantation (ABMT) therapy.

Adenoviridae↗