Biomedical subjects

J S Choi

Publications and source records attributed to J S Choi.

At least 73 records · Page 4Linked to original sources

PubMed1999

Influence of 5,10-methylenetetrahydrofolate reductase gene polymorphism on plasma homocysteine concentration in patients with end-stage renal disease.

The purpose of this study is to observe the influence of the methylenetetrahydrofolate reductase (MTHFR) gene (677C-->T substitution) on plasma homocysteine levels in end-stage renal disease (ESRD) patients who received a relatively large amount of folate (2 mg/d) and are undergoing hemodialysis. A cross-sectional study of plasma homocysteine, vitamin B(12), and folate was performed in patients with ESRD. The study population for the MTHFR gene study included 312 healthy subjects and 106 patients with ESRD undergoing hemodialysis. The C677T transition in the MTHFR gene was detected by HinF 1 restriction enzyme analysis and subsequent electrophoresis in a 3% agarose gel. The genotype of the MTHFR gene in 106 patients with ESRD was homozygous C677T mutation (VV) in 17 patients (16.1%) and heterozygous (AV) in 63 patients (58.4%); 26 patients (24.5%) did not carry this mutation (AA). The mean levels of homocysteine, vitamin B(12), and folate in the patients with ESRD were 23.3 +/- 14.0 mmol/L, 620.2 +/- 98.5 pmol/L, and 138.6 +/- 55.6 nmol/L, respectively. There was no significant difference in homocysteine levels among the three genotypes: 28.2 +/- 19.4 mmol/L for VV, 22.7 +/- 14.9 mmol/L for AV, and 23.4 +/- 11.1 mmol/L for AA genotype (P > 0.05). There was no difference in genotype distribution between the patient groups of less than 25th and greater than 75th percentiles, classified according to plasma homocysteine levels (P = 0.47). In conclusion, with high-dose folate supplementation, the hyperhomocysteinemia in patients with ESRD does not seem to be caused by the 677C-->T mutation in the MTHFR gene.

Cross-Sectional Studies↗

PubMed1999

Effect of resection velocity and suction ring on corneal flap formation in laser in situ keratomileusis.

PURPOSE: To determine the effects of microkeratome-cutting velocities and the suction ring on corneal flap creation. SETTING: Department of Ophthalmology, College of Medicine, The Catholic University of Korea, Seoul, Korea. METHODS: Procedures were performed under clinical settings in 65 pig eyes. Keratometric diopters (D) were measured before and after application of the suction ring. Corneal thickness was measured before and after resection cutting, and the difference was taken as flap thickness. The microkeratome was initially set at a resection thickness of 160 microns. The blade oscillation (turbine velocity) was set at 30, 35, and 42 psi. The translational velocity was arbitrarily divided into high (1 to 2 seconds), moderate (3 to 5 seconds), and low (6 seconds or more). Data were analyzed by varying the velocities consecutively. Photographs of the cut surface were acquired by scanning electron microscopy to evaluate resection morphology by comparison. RESULTS: Mean refractive powers were 39.94 D +/- 0.66 (SD) and 39.69 +/- 0.98 D before and after application of the suction ring, respectively; however, no significant difference was observed (P = .216). Lower surgeon translational velocity resulted in a significantly thicker corneal flap in 8 of the 9 paired comparisons (P < .05). Higher turbine velocity resulted in a significantly thicker corneal flap in 7 of the 9 paired comparisons (P < .05). The cut surface was smoother at higher turbine and lower translational velocity. The initial cut margin was steeper at higher translational velocity. CONCLUSIONS: The increase in intraocular pressure induced by the suction ring had no significant effect on keratometric power. At higher turbine and lower translational velocities, the corneal flap was thicker and the cut surface smoother. Higher translational velocities made the initial cut margin steeper.

Animals↗

PubMed1999

A novel angiogenic factor derived from Aloe vera gel: beta-sitosterol, a plant sterol.

Aloe vera gel has a beneficial effect on wound healing. Because angiogenesis is an essential process in wound healing, we hypothesized that Aloe vera gel might contain potent angiogenic compounds. Here we demonstrate that Aloe vera gel and its extracts are angiogenic on the chorioallantoic membrane (CAM) of chick embryo. Out of the three compounds purified from the final fraction of Aloe vera gel, beta-sitosterol showed a potent angiogenic activity in the CAM assay. In the presence of heparin, beta-sitosterol stimulated neovascularization in the mouse Matrigel plug assay and the motility of human umbilical vein endothelial cells in an in vitro wound migration assay. Thus beta-sitosterol is a novel plant-derived angiogenic factor which may have potential pharmaceutical applications for the management of chronic wounds.

Journal Article↗

PubMed1999

Inhibitory effect of luteolin 4'-O-glucoside from Kummerowia striata and other flavonoids on interleukin-5 bioactivity.

Interleukin (IL)-5 is a chemotactic factor of eosinophils, and promotes the growth and survival of eosinophils, which plays an important role in the eosinophilia-associated allergic inflammation. In this study, luteolin 4'-O-glucopyranoside was identified as the IL-5 inhibitor from Kummerowia striata Thunb. (Leguminosae) by activity-guided fractionation followed by structural analysis compared with reported spectral data. The flavone compound exhibited dose-dependent inhibitory effect on IL-5 bioactivity with 95% inhibition at 30 microM, 79% at 15 microM, 60% at 7.5 microM, 54% at 3.8 microM and 29% at 1.9 microM, where 50% of inhibition (IC50) value was shown at the concentration of 3.7 microM. Furthermore, the inhibitory effect on IL-5 bioactivity by other flavonoid compounds available was estimated. In view of the IC50 values, the inhibitory potency on IL-5 bioactivity was in order of luteolin 4'-O-glucopyranoside > cosmosiin (14.2 microM) approximately equal to apigenin (16.4 microM) approximately equal to luteolin (18.7 microM) > quercimeritrin (27.3 microM) approximately equal to kaempferol (30.0 microM).

Animals↗

PubMed1999

Protective role for bcl-2 in experimentally induced cell death of bovine corneal endothelial cells.

To characterize the pattern of cell death and to investigate the potential role of bcl-2 in a death paradigm of corneal endothelial cells, primary cultures of bovine corneal endothelial (BCEN) cells were first established and treated with 0.01-1 microM staurosporine, a nonspecific protein kinase inhibitor. The pattern of BCEN cell death induced by staurosporine was apoptotic in nature, characterized by shrinkage of the cytoplasmic membrane, nuclear condensation and DNA fragmentation. Cotreatment of BCEN cells with Z-VAD-fmk (a caspase inhibitor) but not cycloheximide (a protein synthesis inhibitor) prevented staurosporine-induced cell death. To investigate the potential role of bcl-2, BCEN cells were transferred with a eukaryotic expression vector containing anti-apoptotic bcl-2 cDNA and characterized by reverse transcription-polymerase chain reaction (RT-PCR; BCEN/bcl-2). As measured by the MTT reduction assay after treatment with staurosporine, the survival rate of BCEN/bcl-2 cells was 48.0 +/- 4.8% compared to 7.4 +/- 2.1% in control BCEN cells. As determined by light microscopy, apoptotic changes such as nuclear condensation and apoptotic bodies were largely attenuated in BCEN/bcl-2 cells after staurosporine treatment although arborization of processes and rounding up of the cell body were not affected by overexpression of bcl-2. These results suggest that staurosporine induces apoptosis in a cycloheximide-independent but caspase-dependent manner and bcl-2 acts as a negative regulator in staurosporine-induced apoptosis of BCEN cells.

Animals↗

PubMed1999

Photomovement of the gliding cyanobacterium Synechocystis sp. PCC 6803.

Using a computerized videomicroscope motion analysis system, we investigated the photomovements of two Synechocystis sp. (PCC 6803 and ATCC 27184). Synechocystis sp. PCC 6803 displays a relatively slow gliding motion. The phototactic and photokinetic speeds of this cyanobacterium in liquid media were 5 microns/min and 15.8 microns/min, respectively, at 3 mumol/m2/s of stimulant white light. Synechocystis sp. PCC 6803 senses light direction rather than intensity for phototaxis. Synechocystis sp. ATCC 27184 showed a weak photokinesis but no phototaxis. Analysis of Synechocystis sp. ATCC 27184 suggests that the loss of phototaxis results from spontaneous mutation during several years of subculture. When directional irradiation was applied, the cell population of Synechocystis sp. PCC 6803 began to deviate from random movement and reached maximum orientation at 5 min after the onset of stimulant white light. Synechocystis sp. PCC 6803 showed high sensitivity to the stimulant white light of fluence rates as low as 0.002 mumol/m2/s. Neither 1,3-dichlorophenyldimethyl urea nor cyanide affected phototactic orientation, whereas cyanide inhibited gliding speed. This result suggests that the phototaxis of Synechocystis sp. PCC 6803 is independent of photosynthetic phosphorylation and that its gliding movement is primarily powered by oxidative phosphorylation. In the visible wavelength region, 560 nm, 660 nm and even 760 nm caused positive phototaxis. However, 360 nm light induced strikingly negative phototaxis. Therefore, at least two independent photoreceptors may exist to control phototaxis. The photoreceptor for positive phototaxis appears likely to be a phytochrome-like tetrapyrrole rather than chlorophyll a.

Agar↗

PubMed1999

Administration-time differences in the pharmacokinetics of gentamicin intravenously delivered to human beings.

Administration-time differences of gentamicin pharmacokinetics were studied by crossover design after a single intravenous administration of gentamicin (80 mg) to 10 human subjects at 09:00 (morning time) and 22:00 (nighttime). The profiles of serum gentamicin concentration showed a significant statistical difference between 09:00 and 22:00, suggesting circadian variations of pharmacokinetic behaviors. A significant circadian rhythm of pharmacokinetic parameters as a function of time of day was noted in human subjects, showing lower total body clearance Clt and higher serum area under the curve (AUC) when given at nighttime. The half-life t1/2 was shorter in the morning (2.82 h +/- 0.43 h) when compared to the nighttime (2.97 h +/- 0.36 h), but the difference was not statistically significant. The AUC was significantly greater in the morning (23.4 +/- 3.84 micrograms-h/mL) than that in the nighttime (26.3 +/- 5.79 micrograms-h/mL) (p < .05), most likely because the Clt was significantly higher when gentamicin was given in the morning (3.51 +/- 0.57 L/h) versus in the nighttime (3.18 +/- 0.65 L/h). Although the volume of distribution Vd decreased when given at nighttime, it was independent of the dosing time. From this study, there was an administration-time difference of gentamicin pharmacokinetics in human beings. The optimized dosing regimen of gentamicin can be decided by considering circadian rhythm and rest-activity routine so that minimized toxicity and effective therapy are established for patients. The current findings also can be applied to other drugs with circadian rhythms of pharmacokinetics and narrow therapeutic windows in clinical chronotherapeutics.

Adult↗

PubMed1999

Laparoscopic splenectomy for immune thrombocytopenic purpura--long-term result of 40 laparoscopic splenectomies.

Laparoscopic surgery has recently extended its indications and it has also become an acceptable surgical approach for splenectomy. In the last five years, we have performed 40 laparoscopic splenectomies for immune thrombocytopenic purpura. Thirty-five patients were female and 5 patients were male. The mean age was 34, varying from 17 to 56. After learning to perform laparoscopic splenectomy with five ports, we are now usually using three or four ports in a right lateral kidney position. There was no case of conversion to exploratory laparotomy. The mean hospital stay was 7 days. There was no perioperative mortality; but in 2 cases we had postoperative subphrenic abscesses which were successfully managed by catheter drainage. Since undergoing laparoscopic splenectomy, 28 patients (70%) were weaned effectively from their steroid medications. Eight patients (20%) have been on small doses of steroid, and 4 patients (10%) have been on the same doses of steroid with no response. The patient group with rapidly increasing platelet count after splenectomy showed a statistically significant relation with the complete response group (p < 0.001). Laparoscopic splenectomy is a safe and reasonable operative procedure for patients with immune thrombocytopenic purpura.

Adolescent↗

PubMed1999

Imported tertian malaria resistant to primaquine.

In Plasmodium vivax and Plasmodium ovale malaria, some of the liver stage parasites remain dormant. The activation of these dormant forms (called hypnozoite) can give rise to relapse weeks, months or years after the initial infection. To prevent relapses, a course of primaquine may be given as terminal prophylaxis to patients. Different strains of Plasmodium vivax vary in their sensitivity to primaquine and, recently, cases of relapse of Plasmodium vivax after this standard primaquine therapy were reported from various countries. We reported a case of primaquine resistant malaria which initially was thought to be relapsed caused by loss of terminal prophylaxis.

Animals↗

PubMed1999

Necrosis and apoptosis after retinal ischemia: involvement of NMDA-mediated excitotoxicity and p53.

PURPOSE: Accumulated evidence has shown that apoptosis and necrosis contribute to neuronal death after ischemia. The present study was performed to study the temporal and spatial patterns of neuronal necrosis and apoptosis after ischemia in retina and to outline mechanisms underlying necrosis and apoptosis. METHODS: Retinal ischemia was induced by increasing intraocular pressure to a range of 160 mm Hg to 180 mm Hg for 90 minutes in adult rats. The patterns of neuronal cell death were determined using light and electron microscopy and were visualized by TdT-dUTP nick-end labeling (TUNEL). The mRNA expression profile of p53 was examined using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization histochemistry. Immunohistochemistry was performed using anti-p53, anti-microtubule associated protein-2, and anti-glial fibrillary acidic protein antibodies. RESULTS: Within 4 hours after ischemia, neurons in the inner nuclear cell layer (INL) and ganglion cell layer (GCL) underwent marked necrosis, made apparent by swelling of the cell body and mitochondria, early fenestration of the plasma membrane, and irregularly scattered condensation of nuclear chromatin. After 3 days, the INL and GCL neurons showed further degeneration through apoptosis marked by cell body shrinkage, aggregation, and condensation of nuclear chromatin. Apoptotic neurons were also observed sparsely in the outer nuclear cell layer. Intravitreal injections of MK-801 prevented early neuronal degeneration after ischemia. Of note, mRNA and protein levels of p53, the tumor suppressor gene known to induce apoptosis, were increased in the retinal areas undergoing apoptosis 1 to 3 days after ischemic injury. CONCLUSIONS: Ischemia produces the N-methyl-D-aspartate-mediated necrosis and slowly evolving apoptosis of neurons in the retina. The latter may depend on the expression of the p53 proapoptosis gene.

Animals↗

PubMed1999

Reactive oxygen species-induced apoptosis and necrosis in bovine corneal endothelial cells.

PURPOSE: The loss of corneal endothelial cells associated with aging and possibly other causes has been speculated to be related to exposure to reactive oxygen species (ROS). The current study was conducted to investigate, by use of photosensitizers, the underlying mechanisms involved in the death of bovine corneal endothelial cells (BCENs) caused by ROS. METHODS: BCEN cells in primary culture were treated with a photosensitizer (riboflavin or rose bengal) with light exposure. The patterns of cell damage and death were assessed using an acridine orange-ethidium bromide differential staining method, TdT-mediated dUTP nick-end labeling (TUNEL) assay, and transmission electron microscopy. The cytotoxicity was assayed by mitochondrial function using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) testing. Antioxidants, including catalase, L-histidine, salicylic acid, and superoxide dismutase, were used to determine the types of ROS involved. Activation of nuclear factor (NF)-kappaB was examined by fluorescent immunocytochemistry with anti-p65 antibody. RESULTS: Light-irradiated riboflavin or rose bengal resulted in a significant decrease in viability of BCEN cells. Chromosomal condensation and fragmentation were observed in apoptotic cells, and membrane lysis and damage of cell ultrastructures were observed in necrotic cells. Riboflavin induced apoptosis at 30 minutes and thereafter and induced necrosis after 2 hours. Rose bengal was shown to cause similar effects within half the time required for the effects of riboflavin. Catalase and salicylic acid were found to provide protection for BCENs from cytotoxic effects of riboflavin, and L-histidine was found to protect BCENs from cytotoxicity induced by rose bengal. Kinetic studies using immunocytochemistry showed that NF-kappaB was translocated into the nucleus within 15 minutes and 30 minutes after treatment with rose bengal and riboflavin, respectively. CONCLUSIONS: The cytotoxic effects of photo-irradiated riboflavin and rose bengal are shown to be mediated by two distinct but parallel pathways, one leading to apoptosis and the other to necrosis. Possible involvement of NF-kappaB in cell death is suggested. These findings provide potential leads for future investigation into the molecular mechanisms of loss of corneal endothelial cells related to aging, oxidative stress, and possibly other similar causes.

Acridine Orange↗

PubMed1999

Circadian changes in pharmacokinetics of cyclosporin in healthy volunteers.

Circadian changes in the pharmacokinetics of cyclosporin were evaluated after oral administration of the drug, 300 mg, at 09:00 and 20:00 to twelve healthy male subjects. At 09:00, the blood concentrations of cyclosporin were higher, the area under the blood concentration-time curve from time 0 to time infinity was significantly greater (9.70 versus 8.42 microg x hr/ml), and the renal clearance was significantly slower (1.14 versus 1.48 ml/min) than those at 22:00. The terminal half-life, peak concentration, and time to reach peak concentration were not significantly different between two groups.

Administration, Oral↗

PubMed1999

Mechanism of fluoxetine block of cloned voltage-activated potassium channel Kv1.3.

The effects of fluoxetine (Prozac), a widely used antidepressant drug, on Kv1.3 stably expressed in Chinese hamster ovary cells were examined using the whole-cell and excised inside-out configurations of the patch-clamp technique. In whole-cell recordings, fluoxetine accelerated the decay rate of inactivation of Kv1.3 and thus decreased the current amplitude at the end of the pulse in a concentration-dependent manner with an IC(50) value of 5.9 microM. The inhibition displayed a weak voltage dependence, increasing at more positive potentials. Neither the activation nor the steady-state inactivation curve was affected by fluoxetine. In addition, fluoxetine reduced the tail current amplitude and slowed the deactivation of the tail current, resulting in a crossover phenomenon. When applied to the internal side of the membrane in inside-out recordings, the inhibition by fluoxetine was much faster and more potent with an IC(50) value of 1.7 microM compared with whole-cell recordings. Norfluoxetine, the major metabolite of fluoxetine, also inhibited Kv1.3 in a concentration-dependent manner (IC(50) = 1.4 microM) in whole-cell recordings. To check whether the fluoxetine-induced inhibition demonstrated in cloned Kv1.3 could also be observed in native T lymphocytes, the effects of fluoxetine were investigated on human T lymphocytes. Fluoxetine also inhibited outward K(+) current in human T lymphocytes. Our results indicate that fluoxetine produced a concentration- and voltage-dependent inhibition of Kv1.3 that can be interpreted as an open channel block and that a binding site for fluoxetine is more accessible from the intracellular side.

Animals↗

PubMed1999

Enhanced secretory group II PLA2 activity in the tears of chronic blepharitis patients.

PURPOSE: Phospholipase A2 (PLA2) hydrolyzes phospholipids, one of the important constituents of human meibomian gland secretions. This study was performed to investigate PLA2 type and activity in the tears of chronic blepharitis patients compared to those of normal persons. METHODS: Tear samples of 36 patients and 10 normal persons were collected in non-heparinized microcapillary tubes. PLA2 activity in the tears was measured by Dole's method, and the results of the blepharitis patients were compared to those of the normal persons. The characterization of PLA2 was performed by the head group preference test and the dithiothreitol (DTT) sensitivity test. The classification of PLA2 type was done using Western blot analysis with anti-human secretory PLA2 antibody. RESULTS: No statistically significant differences were found among the six categories of chronic blepharitis. However, the mean PLA2 activity in the tears of the chronic blepharitis patients was about two times higher than that of the normal controls with statistical significance (P < 0.05). The PLA2 substrate specificity test revealed group II PLA2 activity. Furthermore, the group II PLA2 was identified as a 14 kDa band in Western blot analysis using an antibody raised against human secretory group II PLA2. CONCLUSIONS: Secretory group II PLA2 activity was significantly enhanced in the tears of the chronic blepharitis patients compared with that of the normal controls. It is suggested that this increased enzymatic activity may decrease the tear film stability through increased hydrolysis of phospholipids.

Adult↗

PubMed1998

In vitro activation of pro-phenol-oxidase by two kinds of pro-phenol-oxidase-activating factors isolated from hemolymph of coleopteran, Holotrichia diomphalia larvae.

Previously, we purified and characterized a pro-phenol-oxidase (pro-PO) of 79 kDa from coleopteran insect, Holotrichia diomphalia larvae [Kwon et al. (1997) Mol. Cells 7, 90-97]. Here, we describe the identification of two pro-PO-activating factors (PPAF), named PPAF-I and PPAF-II, directly involved in the activation of the isolated pro-PO. When pro-PO was incubated with either PPAF-I or PPAF-II, no phenol oxidase activity was observed. However, incubation of pro-PO with both PPAF-I and PPAF-II specifically exhibited phenol oxidase activity. The purified PPAF-I with a molecular mass of 33 kDa on SDS/PAGE had characteristics of a serine protease. It exhibited amidase activity against fluorogenic peptide substrates, tert-butoxycarbonyl-phenylalanyl-seryl-arginyl-4-methylcoumaryl-7-amide being the best among the substrates examined. The activity was completely inhibited by 0.02 mM p-nitrophenyl-p'-guanidinobenzoate HCl and diisopropylflurophosphate. The NH2-terminal sequence of PPAF-I had significant sequence similarity to those of serine proteases. On the other hand, the purified PPAF-II had a molecular mass of 40 kDa on SDS/PAGE and 400 kDa determined by gel filtration, indicating an oligomeric protein. The NH2-terminal sequence of PPAF-II showed no similarity to known proteins. PPAF-II exhibited no amidase activity against the fluorogenic substrates. Reconstitution experiments and immunoblotting analysis using affinity-purified antibody against pro-PO demonstrated that PPAF-I first cleaves the intact pro-PO to an intermediate of 76 kDa with no phenol oxidase activity, and then, PPAF-I converts the intermediate to the active phenol oxidase of 60 kDa in the presence of PPAF-II. These results indicate that the activation of pro-PO system in hemolymph of H. diomphalia larvae is accomplished by at least two activating factors, a serine protease and a protein cofactor.

Amidohydrolases↗

PubMed1998

Structure-related inhibition of human hepatic caffeine N3-demethylation by naturally occurring flavonoids.

The effects of flavonoids on caffeine N3-demethylation, a marker activity of CYP1A2, in human liver microsomes were investigated to elucidate the inhibition mechanism and the structure-activity relationship. Caffeine N3-demethylase activity was inhibited by the presence of various flavonoids, whose structures seem to be closely related to the degree of inhibition. Among twenty-one compounds tested, the most active was chrysin with an IC50 value of 0.2 microM. Others had IC50 values ranging from 1 to more than 500 microM. Kinetic analysis revealed that the mechanism of inhibition varied among the flavonoids. The inhibitory effect was postulated to be governed by factors such as the number of hydroxyl groups and glycosylation of these free hydroxyl groups. An increase in the number of free hydroxyl groups reduced the inhibitory effect on P450 activity. Analysis of the quantitative structure-activity relationship (QSAR) showed that the volume to surface area ratio was the most effective factor on the inhibition of caffeine N3-demethylation, and the electron densities on the C3 and C4' atoms exercised significant influence on the inhibitory effect. The calculated inhibitory effect of flavonoids on CYP1A2 activity was highly correlated with the antimutagenicity of flavonoids in 2-amino-3,4-dimethylimidazo[4,5-flquinoline (MelQ)-induced umu response.

Anticarcinogenic Agents↗

PubMed1998

Organization and transcriptional characterization of the cat1 gene cluster in Acinetobacter lwoffi K24.

Previously, we have reported that two clustered cat genes from Acenitobacter lwoffi K24 had different arrangements, catB1C1A1 and catB2A2C2 (Kim, S.I., S.-H. Leem, J.-S. Choi, Y.H. Chung, S. Kim, Y.-M. Park, Y.K. Park, Y.N. Lee, and K.-S. Ha. 1997, J. Bacteriol. 179, 5226-5231). By further analysis of the organization of the cat1 gene cluster, we obtained a complete sequence of the catB1 gene, which encoded 40.8-kDa polypeptide containing 379 amino acids, and found a open reading frame (ORF) coding a putative regulatory protein in upstream region of catB1 on plasmid pCD1-1. This ORF encoded 34.2-kDa polypeptide containing 379 amino acids and had more than 40% identity with catR, LysR family regulatory protein of Pseudomonas putida. RT-PCR, Northern blot analysis and primer extension assay for transcriptional analysis of the cat1 gene cluster revealed that the catB1C1 genes were cotranscribed and the catA1 gene was independently transcribed.

Acinetobacter↗

PubMed1998

Two hevein homologs isolated from the seed of Pharbitis nil L. exhibit potent antifungal activity.

Two antifungal peptides (Pn-AMP1 and Pn-AMP2) have been purified to homogeneity from seeds of Pharbitis nil. The amino acid sequences of Pn-AMP1 (41 amino acid0 residues) and Pn-AMP2 (40 amino acid residues) were identical except that Pn-AMP1 has an additional serine residue at the carboxyl-terminus. The molecular masses of Pn-AMP1 and Pn-AMP2 were confirmed as 4299.7 and 4213.2 Da, respectively. Both the Pn-AMPs were highly basic (pI 12.02) and had characteristics of cysteine/glycine rich chitin-binding domain. Pn-AMPs exhibited potent antifungal activity against both chitin-containing and non-chitin-containing fungi in the cell wall. Concentrations required for 50% inhibition of fungal growth were ranged from 3 to 26 micrograms/ml for Pn-AMP1 and from 0.6 to 75 micrograms/ml for Pn-AMP2. The Pn-AMPs penetrated very rapidly into fungal hyphae and localized at septum and hyphal tips of fungi, which caused burst of hyphal tips. Burst of hyphae resulted in disruption of the fungal membrane and leakage of the cytoplasmic materials. To our knowledge, Pn-AMPs are the first hevein-like proteins that show similar fungicidal effects as thionins do.

Amino Acid Sequence↗