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K T Kim

Publications and source records attributed to K T Kim.

At least 55 records · Page 3Linked to original sources
PubMed1999

Phospholipase C-delta1 is activated by capacitative calcium entry that follows phospholipase C-beta activation upon bradykinin stimulation.

To characterize the regulatory mechanism of phospholipase C-delta1 (PLC-delta1) in the bradykinin (BK) receptor-mediated signaling pathway, we used a clone of PC12 cells, which stably overexpress PLC-delta1 (PC12-D1). Stimulation with BK induced a significantly higher Ca(2+) elevation and inositol 1,4,5-trisphosphate (IP(3)) production with a much lower half-maximal effective concentration (EC(50)) of BK in PC12-D1 cells than in wild type (PC12-W) or vector-transfected (PC12-V) cells. However, BK-induced intracellular Ca(2+) release and IP(3) generation was similar between PC12-V and PC12-D1 cells in the absence of extracellular Ca(2+), suggesting that the availability of extracellular Ca(2+) is essential to the activation of PLC-delta1. When PC12-D1 cells were treated with agents that induce Ca(2+) influx, more IP(3) was produced, suggesting that the Ca(2+) entry induces IP(3) production in PC12-D1 cells. Furthermore, the additional IP(3) production after BK-induced capacitative calcium entry was detected in PC12-D1 cells, suggesting that PLC-delta1 is mainly activated by capacitative calcium entry. When cells were stimulated with BK in the presence of extracellular Ca(2+), [(3)H]norepinephrine secretion was much greater from PC12-D1 cells than from PC12-V cells. Our results suggest that PLC-delta1 is activated by capacitative calcium entry following the activation of PLC-beta, additively inducing IP(3) production and Ca(2+) rise in BK-stimulated PC12 cells.

Animals↗
PubMed1999

Temperature sensitivity of catecholamine secretion and ion fluxes in bovine adrenal chromaffin cells.

The effects of temperature on ion fluxes and catecholamine secretion that are mediated by nicotinic acetylcholine receptors (nAChRs), voltage-sensitive calcium channels (VSCCs), and voltage-sensitive sodium channels (VSSCs) were investigated using bovine adrenal chromaffin cells. When the chromaffin cells were stimulated with DMPP, a nicotinic cholinergic agonist, or 50 mM K+, the intracellular calcium ([Ca2+]i) elevation reached a peak and decreased more slowly at lower temperatures. The DMPP-induced responses were more sensitive to temperature changes compared to high K+-induced ones. In the measurement of intracellular sodium concentrations ([Na+]i), it was found that nicotinic stimulation required a longer time to attain the maximal level of [Na+]i at lower temperatures. In addition, the VSSCs-mediated [Na+]i increase evoked by veratridine was also reduced as the temperature decreased. The measurement of [3H]norepinephrine (NE) secretion showed that the secretion within the first 3 min evoked by DMPP or high K+ was greatest at 37 degrees C. However, at 25 degrees C, the secretion evoked by DMPP, but not that by the 50 mM K+, was greater after 10 min of stimulation. This data suggest that temperature differentially affects the activity of nAChRs, VSCCs, and VSSCs, resulting in differential [Na+]i and [Ca2+]i elevation, and in the [3H]NE secretion by adrenal chromaffin cells.

Adrenal Glands↗
PubMed1999

Signalling pathway leading to an activation of mitogen-activated protein kinase by stimulating M3 muscarinic receptor.

The signalling pathway leading to an activation of mitogen-activated protein (MAP) kinase subtypes Erk-1 and -2 upon stimulation of muscarinic receptor with carbachol in human neuroblastoma SK-N-BE2(C) cells was investigated. Carbachol activated Erk-1/-2 by stimulating M3 muscarinic receptor, as determined by specific antagonists for individual muscarinic receptors. The activation of Erk-1/-2 by carbachol was blocked by the inhibition or down-regulation of protein kinase C (PKC). Among the multiple PKC isoforms expressed in SK-N-BE2(C) cells, only PKCepsilon was activated by the treatment of carbachol, and selective down-regulation of PKCepsilon was sufficient to block Erk-1/-2 activation. Carbachol treatment induced activation of the serine/threonine protein kinase Raf, and an inhibition of Raf blocked Erk-1/-2 activation. Ectopic expression of inhibitory small GTPase Ras, RasN17, blocked the carbachol-induced Raf activation without affecting the activation of PKCepsilon, while the inhibition of PKC blocked the Raf activation. Thus, these results suggest that carbachol-induced activation of PKCepsilon mediates Erk-1/-2 activation by a sequential activation of Ras, Raf and MAP kinase kinase.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMed1999

Relation of latex-specific IgE titer and symptoms in patients allergic to latex.

BACKGROUND: The relation between latex-specific IgE titer and the type or total number of latex-induced symptoms has not been previously investigated. OBJECTIVE: We sought to determine the association of latex-IgE in vitro assay results with the type, number, or severity of symptoms in patients allergic to latex. METHODS: Ninety-one patients with positive histories and positive skin test responses were identified as having type I allergy. Data was collected for reported symptoms after latex exposure. Symptom severity was quantitated by 2 scores: (1) the total number of symptoms to latex exposure and (2) a symptom severity score (3 = anaphylaxis, 2 = asthma, and 1 = rhinoconjunctivitis, urticaria, or both). All subjects underwent AlaSTAT and CAP tests. RESULTS: AlaSTAT class was associated with total number of symptoms (r = 0.32, P <.001) and severity score (r = 0.33, P <.002). Similarly, CAP class was associated with both number of symptoms (r = 0.32, P <. 0001) and severity score (r = 0.31, P <.001). Among the symptoms reported, asthma had the strongest association with a positive in vitro IgE assay (odds ratio = 6.7 [95% confidence interval = 1.9, 25. 6]), followed by urticaria (odds ratio = 1.9 [95% confidence interval = 0.8, 4.6]). Contact dermatitis had no statistical association with in vitro assays in this study. AlaSTAT and CAP class correlated together significantly (r = 0.58, P <.001). CONCLUSION: Patients allergic to latex with higher AlaSTAT or CAP class were more symptomatic. Increasing class or titer also predicted more severe symptoms. Higher class of either the AlaSTAT or CAP assay to latex was strongly associated with latex-related asthma and urticaria and marginally associated with latex-related rhinoconjunctivitis.

Anaphylaxis↗
PubMed1999

Rapid and simple measurement of serotonin N-acetyltransferase activity by liquid biphasic diffusion assay.

We report here a rapid, simple, and accurate method to assay for serotonin N-acetyltransferase (NAT) activity. This assay relies on the selective diffusion of radiolabeled acetyltryptamine into a water-immiscible scintillation fluid. Unlike organic solvent extraction, thin-layer chromatography, or high performance liquid chromatography, the separation of acetyltryptamine from acetyl CoA and tryptamine is not required in the method. Moreover, the limit of sensitivity is less than 4 pmol of N-acetyltryptamine formed per sample. Enhancement of NAT activity upon beta-adrenergic receptor stimulation in the rat pineal gland was clearly detected with this method. In addition, the NAT activity measurements obtained with this method agreed quantitatively in the pineal gland and other brain tissues with the conventional organic solvent extraction method. The results suggest that this liquid biphasic diffusion assay is applicable to the detection of NAT activity in tissues and cells.

Adrenergic beta-Agonists↗
PubMed1999

Pharmacological characterization of beta2-adrenoceptor in PGT-beta mouse pineal gland tumour cells.

1. The adrenoceptor in a mouse pineal gland tumour cell line (PGT-beta) was identified and characterized using pharmacological and physiological approaches. 2. Adrenaline and noradrenaline, adrenoceptor agonists, stimulated cyclic AMP generation in a concentration-dependent manner, but had no effect on inositol 1,4,5-trisphosphate production. Adrenaline was a more potent activator of cyclic AMP generation than noradrenaline, with half maximal-effective concentrations (EC50) seen at 175+/-22 nM and 18+/-2 microM for adrenaline and noradrenaline, respectively. 3. The addition of forskolin synergistically stimulated the adrenaline-mediated cyclic AMP generation in a concentration-dependent manner. 4. The pA2 value for the specific beta2-adrenoceptor antagonist ICI-118,551 (8.7+/-0.4) as an antagonist of the adrenaline-stimulated cyclic AMP generation were 3 units higher than the value for the betaI-adrenoceptor antagonist atenolol (5.6+/-0.3). 5. Treatment of the cells with adrenaline and forskolin evoked a 3 fold increase in the activity of serotonin N-acetyltransferase with the peak occurring 6 h after stimulation. 6. These results suggest the presence of beta2-adrenoceptors in mouse pineal cells and a functional relationship between the adenylyl cyclase system and the regulation of N-acetyltransferase expression.

Adenylyl Cyclases↗
PubMed1999

Characterization of high affinity neurotensin receptor NTR1 in HL-60 cells and its down regulation during granulocytic differentiation.

1. We investigated responses to neurotensin in human promyelocytic leukaemia HL-60 cells. 2. Neurotensin increased the cytosolic calcium concentration ([Ca2+]i) in a concentration-dependent manner and also produced inositol 1,4,5-trisphosphate (InsP3). 3. Among the tested neurotensin analogues, neurotensin 8-13, neuromedin-N, and xenopsin also increased [Ca2+]i, whereas neurotensin 1-11 and neurotensin 1-8 did not elicit detectable responses. 4. SR48692, an antagonist of NTR1 neurotensin receptors, blocked the neurotensin-induced [Ca2+]i increase, whereas levocabastine, which is known as an NTR2 neurotensin receptor antagonist, did not attenuate the neurotensin-evoked effect. 5. The expression of NTR1 neurotensin receptors was confirmed by Northern blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR). 6. During 1.25% dimethylsulfoxide (DMSO)-triggered granulocytic differentiation of HL-60 cells, the neurotensin-induced [Ca2+]i rise became gradually smaller and completely disappeared 4 days after treatment with DMSO. The mRNA level for neurotensin receptors was also decreased after differentiation. 7. The results show that HL-60 cells express NTR1 neurotensin receptors and suggest that granulocytic differentiation involves transcriptional regulation of the receptors resulting in down-regulation of the neurotensin-induced signalling.

Calcium↗
PubMed1999

Orbital floor and infraorbital rim reconstruction after total maxillectomy using a vascularized calvarial bone flap.

A number of techniques have been introduced to support the orbital floor after maxillectomy without orbital exenteration. These methods include skin graft or muscular sling, but they have resulted in severe complications, such as enophthalmos, global ptosis, diplopia, and facial deformity. Currently, advanced microvascular reconstruction using bone and soft tissue is performed by many surgeons. This usually results in the filling of the postmaxillectomy defect, but the lack of support for the orbital rim and floor by the bone flap may still cause the complications mentioned above. Vascularized calvarial bone flap was chosen in this study for reconstruction of the orbital floor and infraorbital rim to function as a buttress, to reconstruct recipient sites of poor vascular bed after radiation therapy, and to withstand further postoperative radiation. By providing a solid floor and rim, these complications can be prevented with satisfactory function and aesthetically acceptable results. From September of 1995 to July of 1998, we performed vascularized bone flap for the reconstruction of the orbital floor and infraorbital rim in four cases after total maxillectomy involving the orbital floor. With a follow-up period from 19 to 35 months (mean, 27 months), we obtained significant improvement of functional and aesthetically acceptable results without global ptosis, enophthalmos, diplopia, or severe facial contour deformity.

Aged↗
PubMed1999

Chronic pruritic papular dermatitis in adult men: a variant of prurigo.

Chronic recurrent pruritic papular eruptions in which a specific diagnosis cannot be established becomes a baffling experience to the dermatologist. We have met adult male patients with chronic recurrent pruritic papular eruptions, but their clinicopathological features are not described in English language textbooks. Our purpose was to study the clinical and histological features of this entity and review the various existing taxonomy. We conducted a study of 20 patients over a six year period by taking histories, performing skin biopsies, screening patch tests, and doing immunofluoresence studies. The eruptions occurred exclusively in male adults and had a predilection for the trunk and proximal extremities. The lesions were characterized by severely pruritic, nonfollicular, monomorphic, erythematous urticarial papules. There was no evidence of atopic diathesis or history of insect bite. Most patients had normal levels of serum eosinophils and IgE. The predominant histopathologic finding was a presence of perivascular infiltration of mononuclear cells with eosinophils. The patients followed a chronic course of at least six months with waxing and waning; systemic corticosteroids were the only effective treatment. Finally, all other pruritic erythematous papular dermatoses were ruled out. These cases comprise a distinct entity that has previously been mentioned in a few reports. Clear definition of this entity with an appropriate designation is in order to avoid confusion among dermatologists, and we propose the disease name "chronic papular dermatitis in adult men" as a variant of prurigo.

Adrenal Cortex Hormones↗
PubMed1999

Update on the status of Africanized honey bees in the western states.

The Africanized honey bee (AHB), Apis mellifera scutella--perhaps better known as the "killer bee"--has arrived in the western United States and in southern California, following a nearly 50-year north-ward migration across South and Central America. First detected near Hidalgo, Texas in October 1993, the bees continue to advance 100 to 300 miles per year by colonizing existing hives or forming new hives in the wild. Although the AHB's "killer" reputation has been greatly exaggerated, the presence of AHBs will increase the chances of people being stung.

Animals↗
PubMed1999

Capsaicin inhibits phospholipase C-mediated Ca(2+) increase by blocking thapsigargin-sensitive store-operated Ca(2+) entry in PC12 cells.

Capsaicin has been shown to act through vanilloid receptors, which are temperature-sensitive cation channels. However, there also are indications that suggest the capsaicin effect is not mediated by the vanilloid receptor. We therefore investigated the effect of capsaicin on the phospholipase C-mediated Ca(2+) rise in PC12 cells. Capsaicin caused a rapid decline in extracellular ATP- or bradykinin-induced calcium transients to the basal level without significant attenuation of the peak level. However, capsaicin did not inhibit either ATP- or bradykinin-induced Ca(2+) elevation in the absence of extracellular Ca(2+) or inositol-1,4,5-trisphosphate production. Capsaicin also inhibited ATP-induced norepinephrine secretion. Capsaicin dramatically reduced the thapsigargin-induced sustained Ca(2+) level, suggesting that capsaicin inhibits thapsigargin-sensitive store-operated Ca(2+) entry (SOCE). Thapsigargin-induced Ba(2+) and Mn(2+) influx was also inhibited by capsaicin. Furthermore, capsaicin overlapped SK&F96365 in inhibiting thapsigargin-sensitive SOCE. Capsaicin-induced inhibition of SOCE also occurred in thapsigargin-treated Jurkat-T cells, which have a rather prominent SOCE. Resiniferatoxin, a vanilloid receptor agonist, did not mimic the effect of capsaicin. Ruthenium red and capsazepine, which are known to inhibit the vanilloid receptor, did not affect this capsaicin effect. The results suggest that capsaicin does not mediate vanilloid receptor signaling when inhibiting the thapsigargin-sensitive SOCE. The capsaicin action was also not mediated by activation of protein kinase C because phorbol-12-myristate 13-acetate and capsaicin did not overlap each other's effect and GF109203X did not reverse the inhibitory effect of capsaicin. The results suggest that capsaicin negatively modulates thapsigargin-sensitive SOCE subsequent to phospholipase C activation.

Animals↗
PubMed1999

Neomycin inhibits catecholamine secretion by blocking nicotinic acetylcholine receptors in bovine adrenal chromaffin cells.

We investigated the effects of neomycin on nicotinic acetylcholine receptor-induced responses in bovine adrenal chromaffin cells. Neomycin inhibited the nicotinic agonist dimethylphenylpiperazinium iodide (DMPP)-induced norepinephrine secretion in a concentration-dependent manner. Neomycin had also an inhibitory effect on the DMPP-induced increase in cytosolic Ca++ concentration ([Ca++]i). This effect was further confirmed by inhibition of the DMPP-induced fluorescence quenching of fura-2 upon Mn++ entry. Under the same conditions, however, neomycin did not change the bradykinin-induced [Ca++]i increase, which follows the downstream signal of phospholipase C phospholipase C activation in this cell. The inhibitory effect of neomycin on the DMPP-induced [Ca++]i increase was apparent when the neomycin treatment was performed simultaneously with DMPP, suggesting a direct action on the nicotinic receptor. The direct inhibitory action of neomycin on the nicotinic receptor was also evident when neomycin inhibited the DMPP-induced cytosolic Ca++ increase, which is not affected by nifedipine nor omega-conotoxin MVIIC, and the cytosolic Na+ increase, which is not affected by tetrodotoxin. In addition, we observed that neomycin inhibited the binding of nicotine to the acetylcholine receptor in a noncompetitive manner. The data suggest that neomycin inhibits the nicotinic acetylcholine receptor directly, which results in blockage of the nicotinic receptor-mediated signaling without involvement of phospholipase C.

Adrenal Glands↗
PubMed1998

Opposing effects of protein kinase A and C on capacitative calcium entry into HL-60 promyelocytes.

Treatment of HL-60 cells with thapsigargin, a microsomal Ca2+/ATPase inhibitor, led to depletion of intracellular calcium stores followed by capacitative calcium entry. Stimulation of adenylyl cyclase with forskolin enhanced thapsigargin-induced Ca2+ influx. The forskolin effect was confirmed by enhanced fluorescence quenching induced by Mn2+ entry into fura-2 loaded cells. 1,9-Dideoxy-forskolin, an inactive analog of forskolin, did not affect capacitative calcium entry. On the other hand, phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, inhibited thapsigargin-induced Ca2+ entry. Histamine and prostaglandin E2 (PGE2) elevated intracellular adenosine 3':5'-cyclic monophosphate (cAMP) levels and enhanced the thapsigargin-induced capacitative calcium entry. Incubation with N-[2-(p-bromocynnamylamino)ethyl]-5-isoquinolinesulfonamide (H89), an inhibitor of protein kinase A (PKA), blocked the forskolin effect, and GF109203X, an inhibitor of protein kinase C (PKC), blocked the phorbol 12-myristate 13-acetate effect. The results suggest that protein kinase A regulates capacitative calcium entry positively, but that protein kinase C regulates Ca2+ influx negatively. Furthermore, after differentiation of HL-60 promyelocytes with dimethylsulfoxide to granulocytes, the inhibitory effect of phorbol 12-myristate 13-acetate became more pronounced, whereas the stimulatory effect of prostaglandin E2 did not change. This result suggests that the regulation of capacitative calcium entry by protein kinase C and protein kinase A develops differently during differentiation.

Calcium↗
PubMed1998

Characterization of Mas-7-induced pore formation in SK-N-BE(2)C human neuroblastoma cells.

Mastoparan, a peptide toxin from wasp venome, mimics receptors by stimulating the GTPase activity of guanine nucleotide binding proteins and the G-protein-coupled phospholipase C (PLC). By using Mas-7, the active analog of mastoparan, we showed that it makes pores in the plasma membrane. Treatment with Mas-7 but not Mas-17, the inactive analog, produced a concentration-dependent rise in intracellular Ca2+ concentration ([Ca2+]i) and facilitated the uptake of ethidium bromide (EtBr) (314 Da) to a sustained level during the stimulation. In addition, Mas-7 triggered the influx of lucifer yellow (457 Da), while it did not induce the influx of fura-2 (831 Da) and Evans blue (961 Da). However, the Mas-7-induced permeability was selectively prevented by the addition of La3+, Ni2+, and Co2+, but not Cd2+. This blocking activity was concentration-dependent. While the stimulatory effect of Mas-7 on PLC activity was dependent on extracellular Ca2+, the pore forming activity of Mas-7 was not effected by removal of extracellular Ca2+. These results, therefore, suggest that the mastoparan's action in pore formation is independent from its action in PLC stimulation and is negatively effected by inorganic cations.

Calcium↗
PubMed1998

Inhibitory effects of bulbocapnine on dopamine biosynthesis in PC12 cells.

The effects of bulbocapnine, an aporphine isoquinoline alkaloid, on dopamine biosynthesis in PC12 cells were investigated. Bulbocapnine showed 45.2% inhibition on dopamine content in PC12 cells at a concentration of 20 microM for 12 h. The IC50 value of bulbocapnine was 26.7 microM. Bulbocapnine at concentrations up to 80 microM was not cytotoxic towards PC12 cells. Tyrosine hydroxylase (TH) activity was inhibited by the treatment of bulbocapnine in PC12 cells (24.4% inhibition at 20 microM). Bulbocapnine at 20 microM also decreased the intracellular Ca2+ concentration by 12.9% inhibition relative to control in PC12 cells. However, TH mRNA level was not altered by bulbocapnine treatment. These results suggest that the inhibition of TH activity by bulbocapnine might be involved in at least one component of the reduction of dopamine biosynthesis in PC12 cells.

Animals↗
PubMed1998

Differential inhibition of catecholamine secretion by amitriptyline through blockage of nicotinic receptors, sodium channels, and calcium channels in bovine adrenal chromaffin cells.

We investigated the effects of amitriptyline, a tricyclic antidepressant, on [3H]norepinephrine ([3H]NE) secretion and ion flux in bovine adrenal chromaffin cells. Amitriptyline inhibited [3H]NE secretion induced by 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP) and 70 mM K+. The half maximal inhibitory concentration (IC50) was 2 microM and 9 microM, respectively. Amitriptyline also inhibited the elevation of cytosolic calcium ([Ca2+]i) induced by DMPP and 70 mM K+ with IC50 values of 1.1 microM and 35 microM, respectively. The rises in cytosolic sodium ([Na+]i) and [Ca2+]i induced by the Na+ channel activator veratridine were also inhibited by amitriptyline with IC50 values of 7 microM and 30 microM, respectively. These results suggest that amitriptyline at micromolar concentrations inhibits both voltage-sensitive calcium (VSCCs) and sodium channels (VSSCs). Furthermore, submicromolar concentrations of amitriptyline significantly inhibited DMPP-induced [3H]NE secretion and [Ca2+]i rise, but not veratridine- or 70 mM K+-induced responses, suggesting that nicotinic acetylcholine receptors (nAChR) as well as VSCCs and VSSCs can be targeted by amitriptyline. DMPP-induced [Na+]i rise was much more sensitive to amitriptyline than the veratridine-induced rise, suggesting that the influx of Na+ and Ca2+, through the nAChR itself is blocked by amitriptyline. Receptor binding competition analysis showed that binding of [3H]nicotine to chromaffin cells was significantly affected by amitriptyline at submicromolar concentrations. The data suggest that amitriptyline inhibits catecholamine secretion by blocking nAChR, VSSC, and VSCC.

Adrenal Glands↗
PubMed1998

The effect of pyrimidine nucleosides on adenosine-induced apoptosis in HL-60 cells.

Among several nucleosides, adenosine is the only one to induce typical apoptotic cell death in human promyelocytic leukemia HL-60 cells. Intracellularly transported adenosine seemed to be required for the induction of apoptosis, since dipyridamole, which inhibits the transport of adenosine, strongly suppressed apoptosis, and 8-phenyltheophylline, a receptor antagonist, did not affect the adenosine-induced effect. The viability of adenosine-treated HL-60 cells was partially recovered by supplementation with a pyrimidine nucleoside, uridine or thymidine. Cytidine or deoxycytidine had no effect on the growth and survival of adenosine-treated cells, while uridine or thymidine inhibited adenosine-induced intracellular DNA fragmentation. These results suggest that the quantitative adjustment of purine and pyrimidine nucleosides might play an important role in the adenosine-induced apoptosis of HL-60 cells. The reduction of c-Myc expression in adenosine-treated cells was prevented by uridine or thymidine. These observations suggest that the expression of c-Myc might be related to an intracellular sensing system for the quantitative balance of nucleosides or nucleotides.

Adenosine↗