Effect of vacancy disorder in phase-change materials.

Ge-Sb-Te-based phase-change materials (PCMs) exhibit contrasting electrical and optical properties upon change in atomic structures, which contain the octahedral p -orbital bonding and also substantial disordered vacancies. While extensive studies have been carried out, there is little detailed analysis of how the vacancy distribution and bonding nature are inter-correlated to affect the physical properties. We studied the effect of vacancy distribution on the octahedral p -bonding network in PCMs using a simple tight-binding model and ab initio calculations. We showed that the octahedral p -bonding network can be described as a collection of independent linear chains and that the vacancy disorders are rephrased as a distribution of atomic chain pieces. This finding enables to link the vacancy distribution to various aspects of materials properties such as total energy, structural distortions, and charge localization.

Luteolin and luteolin-7-O-glucoside protect against acute liver injury through regulation of inflammatory mediators and antioxidative enzymes in GalN/LPS-induced hepatitic ICR mice.

BACKGROUND/OBJECTIVES: Anti-inflammatory and antioxidative activities of luteolin and luteolin-7-O-glucoside were compared in galactosamine (GalN)/lipopolysaccharide (LPS)-induced hepatitic ICR mice. MATERIALS/METHODS: Male ICR mice (6 weeks old) were divided into 4 groups: normal control, GalN/LPS, luteolin, and luteolin-7-O-glucoside groups. The latter two groups were administered luteolin or luteolin-7-O-glucoside (50 mg/kg BW) daily by gavage for 3 weeks after which hepatitis was induced by intraperitoneal injection of GalN and LPS (1 g/kg BW and 10 µg/kg BW, respectively). RESULTS: GalN/LPS produced acute hepatic injury by a sharp increase in serum AST, ALT, and TNF-α levels, increases that were ameliorated in the experimental groups. In addition, markedly increased expressions of cyclooxygenase (COX)-2 and its transcription factors, nuclear factor (NF)-κB and activator protein (AP)-1, were also significantly attenuated in the experimental groups. Compared to luteolin-7-O-glucoside, luteolin more potently ameliorated the levels of inflammatory mediators. Phase II enzymes levels and NF-E2 p45-related factor (Nrf)-2 activation that were decreased by GalN/LPS were increased by luteolin and luteolin-7-O-glucoside administration. In addition, compared to luteolin, luteolin-7-O-glucoside acted as a more potent inducer of changes in phase II enzymes. Liver histopathology results were consistent with the mediator and enzyme results. CONCLUSION: Luteolin and luteolin-7-O-glucoside protect against GalN/LPS-induced hepatotoxicity through the regulation of inflammatory mediators and phase II enzymes.

Assessment of human estrogen receptor agonistic/antagonistic effects of veterinary drugs used for livestock and farmed fish by OECD in vitro stably transfected transcriptional activation assays.

The presence of veterinary drug residues in foods and the environment could potentially cause adverse effects on humans and wildlife. Several veterinary drugs were reported to exhibit endocrine disrupting effects via binding affinities to sexual hormone receptors such as estrogen and androgen receptors. Therefore, we confirmed the human estrogen receptor (ER) agonistic/antagonistic effects of 135 chemicals that were used as veterinary drugs in Korea by the official Organization for Economic Cooperation and Development (OECD) in vitro ER transcriptional activation (TA) assay using the VM7Luc4E2 cell line. In the case of ER agonist screening, 7 veterinary drugs (cefuroxime, cymiazole, trenbolone, zeranol, phoxim, altrenogest and nandrolone) were determined to be ER agonists. In addition, only zeranol was found to exhibit weak ER antagonistic activity. These 7 veterinary drugs, which were determined as ER agonists and/or antagonists by an OECD in vitro assay, were also found to have binding affinity to ERs. These results indicate that various veterinary drugs possess potential (anti-)estrogenic effects. However, further study is needed to determine the precise endocrine-disrupting effects of these compounds.

Anti-H antibody of unusually high titer showing variable reactivities against group A red cells and broad thermal amplitude in a patient with lymphoma.

We report a case of a patient with high titer anti-H antibody showing broad thermal amplitude and variable reactivities against group A red cells. A 62-year-old Korean female was diagnosed with diffuse large B cell lymphoma involving multiple organs. Her ABO/RhD type was A+ and her genotype was ABO*A.01.01/ABO*O.01.02. Antibody screening test (AST) and antibody identification test (IDT) were strongly positive for all reagent cells. Anti-human globulin (AHG) test revealed an antibody titer of 1:256 for 37 °C phase and trace positivity for poly- and mono-specific C3d. Reactivity was stronger for O+ red cells than that for A+ red cells across all temperatures tested (4 °C, room temperature (RT) and 37 °C). This was also found for AHG phase. Anti-IH was ruled out based on agglutination of O+ cord cells (CCs). Antibody was determined as IgM anti-H after DTT treatment. Three batches of 10 A+ red cells from random donors were tested with three consecutive serums for crossmatching using tube method. Interestingly, out of thirty A+ red cells tested, 20 cells at RT, 11 cells at 37 °C and 11 cells in the AHG phase showed reactivity of greater than 2+. The patient was transfused with 6 units of packed RBCs subsequently. Chemotherapy (R-CHOP regimen) and Helicobacter pylori eradication were then started. Her antibody titer gradually decreased following such treatment. In conclusion, we identified a case of patient with high titer anti-H with broad thermal amplitude, suggesting that anti-H antibodies might need to be considered for cases with pan-agglutination in AST and IDT.

Cleavage of cytoplasm within the oligonucleate zoosporangia of allomyces macrogynus.

Allomyces macrogynus produces zoosporangia that discharge uninucleate zoospores after cleavage of multinucleate cytoplasm. Cleavage of cytoplasm within the oligonucleate zoosporangia of A. macrogynus was visualized by constructing three-dimensional models based on electron micrographs and confocal images. In oligonucleate zoosporangia, three adjacent nuclei can form three cleavage planes with a line of intersection of the planes. The position and boundary of the cleavage planes are thought to be determined by the relative positions of the nuclei. The establishment of three cleavage planes by cleavage membranes occurred sequentially, and the nuclear axis connecting the centers of two nuclei affected the development of cleavage membranes on each cleavage plane. In multinucleate zoosporangia, groups of three neighboring nuclei near the cell cortex may initiate the sequential establishment of cleavage planes and then may interact with the nuclei further from the cortex until the interactions of nuclei are propagated to the central region of the cytoplasm.

Luteolin and luteolin-7-O-glucoside inhibit lipopolysaccharide-induced inflammatory responses through modulation of NF-κB/AP-1/PI3K-Akt signaling cascades in RAW 264.7 cells.

Luteolin is a flavonoid found in abundance in celery, green pepper, and dandelions. Previous studies have shown that luteolin is an anti-inflammatory and anti-oxidative agent. In this study, the anti-inflammatory capacity of luteolin and one of its glycosidic forms, luteolin-7-O-glucoside, were compared and their molecular mechanisms of action were analyzed. In lipopolysaccharide (LPS)-activated RAW 264.7 cells, luteolin more potently inhibited the production of nitric oxide (NO) and prostaglandin E2 as well as the expression of their corresponding enzymes (inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) than luteolin-7-O-glucoside. The molecular mechanisms underlying these effects were investigated to determine whether the inflammatory response was related to the transcription factors, nuclear factor (NF)-κB and activator protein (AP)-1, or their upstream signaling molecules, mitogen-activated protein kinases (MAPKs) and phosphoinositide 3-kinase (PI3K). Luteolin attenuated the activation of both transcription factors, NF-κB and AP-1, while luteolin-7-O-glucoside only impeded NF-κB activation. However, both flavonoids inhibited Akt phosphorylation in a dose-dependent manner. Consequently, luteolin more potently ameliorated LPS-induced inflammation than luteolin-7-O-glucoside, which might be attributed to the differentially activated NF-κB/AP-1/PI3K-Akt pathway in RAW 264.7 cells.

Enhancement of the catalytic activity of a 27 kDa subtilisin-like enzyme from Bacillus amyloliquefaciens CH51 by in vitro mutagenesis.

AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. To enhance the catalytic activity of AprE51, two residues, Gly-169 and Ser-101, which, according to the three-dimensional structural model of subtilisin, are located in the P1 substrate-binding site and S3 subsite, respectively, were mutated by site-directed mutagenesis. Results of the mutational analysis showed that substitution of alanine for Gly-169 increased the fibrinolytic activity 1.4-fold. All four Ser-101 mutations, that is, replacements with arginine, leucine, lysine, and tryptophan, also increased the fibrinolytic activity up to 3.9-fold. The S101W mutant with a bulky side chain was more active than mutants with a positively charged or nonpolar small side chains. The fibrinolytic activity of the S101W mutant was further increased by error-prone polymerase chain reaction. The AprE51-6 mutant (S101W/G169A/V192A) had stronger fibrinolytic activity than the S101W mutant. Purified AprE51-6 had a 2.5-fold higher k(cat) and a 2.3-fold lower K(m), which resulted in a 6-fold increase in catalytic efficiency (k(cat)/K(m)) relative to that of wild-type AprE51. In addition, AprE51-6 showed a relatively broader pH range and increased thermostability as compared to AprE51.

Fibrinolytic and antiplatelet aggregation properties of a recombinant Cheonggukjang kinase.

This study characterized the efficacy of recombinant Cheonggukjang kinase (CGK) 3-5-rich fraction as a thrombolytic agent, which we overexpressed in Bacillus licheniformis ATCC10716, a strain normally lacking fibrinolytic activity. We found that CGK3-5 is a plasmin-like protease that directly degrades fibrin clots and does not activate plasminogen during fibrin clot lysis and platelet-rich clot lysis assays. We also confirmed antiplatelet and antithrombotic activity by CGK3-5-rich fraction both in vitro and in vivo. CGK3-5-rich fraction inhibited collagen-induced platelet aggregation in platelet-rich plasma in a concentration-dependent manner. The concentration of 1.5 mg/mL CGK3-5-rich fraction completely inhibited collagen-induced platelet aggregation. Furthermore, injection of CGK3-5-rich fraction into tail veins dose-dependently protected mice from death by pulmonary embolism induced by collagen and epinephrine. The survival rates were 30%, 70%, and 100%, respectively, with doses of 130 mg/kg, 260 mg/kg, and 520 mg/kg. These findings suggest that CGK3-5 holds promise as a treatment to mitigate the potentially effects of stroke and heart failure.

Cytokinesis of the binucleate zoosporangia of Allomyces macrogynus.

Allomyces macrogynus, a true fungus, produces zoosporangia which discharge uninucleate zoospores after cytoplasmic cleavage. Binucleate zoosporangia of A. macrogynus were induced and examined to understand the basic principles of cytokinesis associated with the multinucleate zoosporangia. Development of cleavage membranes was visualized by constructing three dimensional models based on electron micrographs and confocal images. Cleavage membranes on the cleavage plane showed asymmetric ingression from the cortex, but cleavage of cytoplasm was completed by the fusion of cleavage membranes with plasma membrane. Also, the position of the cleavage plane was continuously rotated until settled at the last stage. These studies suggest that the positions of the numerous cleavage planes within a multinucleate zoosporangium are continuously adjusted during development of cleavage membranes. The final settlement of cleavage planes would define the exact boundary of cleavage planes and the expansion of cleavage membranes toward the boundary could complete the cleavage of cytoplasm.

Cloning of aprE86-1 gene encoding 27 kDa mature fibrinolytic enzyme from Bacillus amyloliquefaciens CH86-1.

A gene, encoding the major secreted fibrinolytic protein of Bacillus amyloliquefaciens CH86-1, was cloned from the genomic DNA. DNA sequencing showed that the gene, aprE86-1, could direct the synthesis of a mature protein of 275 amino acids long after processing. When aprE86-1 was introduced into B. subtilis, 27 kDa mature protein was produced as expected. The fibrinolytic activity of B. subtilis transformant (TF) was higher than that of B. amyloliquefaciens CH86-1, showing the possibility of increasing fibrinolytic activities of Bacillus strains through genetic engineering.

Characterization of a 27 kDa fibrinolytic enzyme from Bacillus amyloliquefaciens CH51 isolated from cheonggukjang.

Bacillus amyloliquefancies CH51 isolated from cheonggukjang, a traditional Korean fermented soy food, has strong fibrinolytic activity and produces several fibrinolytic enzymes. Among four different growth media, tryptic soy broth was the best in terms of supporting cell growth and fibrinolytic activity of this strain. A protein with fibrinolytic activity was partially purified from the culture supernatant by CMSephadex and Phenyl Sepharose column chromatographies. Tandem mass spectrometric analysis showed that this protein is a homolog of AprE from B. subtilis and it was accordingly named AprE51. The optimum pH and temperature for partially purified AprE51 activity were 6.0 and 45 degrees , respectively. A gene encoding AprE51, aprE51, was cloned from B. amyloliquefaciens CH51 genomic DNA. The aprE51 gene was overexpressed in heterologous B. subtilis strains deficient in fibrinolytic activity using an E.colo-Bacillus Shuttle vector, pHY300PLK.

Methionine-induced hyperhomocysteinemia modulates lipoprotein profile and oxidative stress but not progression of atherosclerosis in aged apolipoprotein E knockout mice.

It is documented that hyperhomocysteinemia (HHcy) is an independent risk factor for atherosclerosis, but whether elevated plasma homocysteine contributes to the progression of atherosclerosis in aged animals with hypercholesterolemia is still unknown. HHcy was induced in apolipoprotein E (ApoE) knockout mice (male, 32 weeks old) by feeding 2% methionine/low folate (1 mg/kg) diet for 20 weeks. HHcy induced by methionine feeding significantly increased oxidative stress, as measured by thiobarbituric-reactive substances in livers (P < .05) and genetic expression of Cu,Zn-superoxide dismutase, in methionine-fed animals compared with controls (P < .05). Furthermore, lipoprotein profiles were changed, in that low-density lipoprotein-cholesterol was shifted to very low-density lipoprotein in the methionine-supplemented group. However, nuclear factor kappaB activity, atherosclerotic lesions, hepatic glutathione level, lipid profiles, and activities of aspartate aminotransferase and alanine aminotransferase were not significantly different. These findings suggest that HHcy induced by methionine may promote disturbances in lipid peroxidation and modify lipoprotein metabolism but not contribute to the progression of atherosclerotic lesion in aged ApoE knockout mice.

Methionine supplementation accelerates oxidative stress and nuclear factor kappaB activation in livers of C57BL/6 mice.

The present study was designed to investigate whether hyperhomocysteinemia (HHcy) induced by methionine supplementation promotes oxidative stress and nuclear factor kappaB (NFkappaB) activation in livers of C57BL/6 mice when fed a 2% methionine and low folate (1 mg/kg) diet for 12 weeks. Plasma homocysteine concentrations of mice fed methionine were found to be 49 micromol/L by 12 weeks of feeding, which was five times higher than that of controls. HHcy induced by methionine feeding significantly increased oxidative stress, as measured by thiobarbituric acid-reactive substances (P < .05) in livers. This was further confirmed by lower levels of hepatic glutathione (P < .05) and elevated mRNA expressions of hepatic antioxidative enzymes, such as Cu,Zn-superoxide dismutase, catalase, glutathione reductase, and L-gulonolactone oxidase in methionine-fed animals (P < .05). Hepatic function of mice fed methionine seems to be normal, while hepatic triglyceride concentration was lowered by methionine feeding. NFkappaB nuclear binding activities of livers were higher in the methionine group than in the control group. The above results suggest that HHcy induced by methionine may promote disturbances in lipid peroxidation and antioxidant processes and be a pro-inflammatory mediator in livers of C57BL/6 mice.

Effect of bamboo culm extract on oxidative stress and genetic expression: bamboo culm extract ameliorates cell adhesion molecule expression and NFkappaB activity through the suppression of the oxidative stress.

BACKGROUND & AIMS: This study was designed to investigate whether bamboo culm extract (BCE) supplementation may ameliorate risk factors of cardiovascular diseases, such as hypercholesterolemia. METHODS: Oxidative stress and inflammatory mediators in plasma, livers of C57BL/6 mice fed high-cholesterol diet and calf pulmonary artery endothelial (CPAE) cells. Briefly, C57BL/6 mice were fed the high-cholesterol diet which was supplemented with 1% (w/w), or 3% (w/w) of BCE for 16 weeks. The concentration of total cholesterol, LDL-cholesterol, HDL-cholesterol level and atherogenic index were measured. Plasma TEAC value, hepatic thiobarbituric acid reactive substances (TBARS), protein carbonyl values and hepatic antioxidant enzyme activities, such as Cu,Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase (GSH-Px), GSH reductase and catalase were determined. In addition, hepatic nuclear factor kappa B activities were detected. In the calf pulmonary artery endothelial (CPAE) cells stimulated with lipopolysaccharide, the expression of vascular cell adhesion molecule-1 (VCAM-1) and intracellular cell adhesion molecule-1 (ICAM-1) were measured. RESULTS: Plasma cholesterol level was decreased, while HDL-cholesterol was increased, thus atherogenic index was lowered in BCE-supplemented animals. Plasma trolox equivalent and hepatic thiobarbituric acid reactive substances and protein carbonyl values were lowered significantly in BCE groups (p<0.05) in a dose-dependent manner. Hepatic antioxidative enzyme activities, such as Cu,Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase (GSH-P), GSH reductase, and catalase were elevated in mice fed BCE-supplemented diets (p<0.05). Nuclear factor kappa B activities of livers and vascular cell adhesion molecule-1 and intracellular cell adhesion molecule-1 expressions in CPAE cells stimulated with lipopolysaccharide were significantly lowered in BCE groups (p<0.05). CONCLUSION: These results suggest that BCE supplementation may modulate lipoprotein composition and attenuate oxidative stress by elevated antioxidative processes, thus suppressing inflammatory mediator activation as possible mechanism of its anti-atherogenic effect.

Protective effect of pinitol against D-galactosamine-induced hepatotoxicity in rats fed on a high-fat diet.

The protective effect of pinitol against D-galactosamine (GalN)-induced liver damage was examined. Forty male Sprague-Dawley rats were divided into normal control, GalN control, and pinitol groups (0.5%, 1%, and 2%). After 8 weeks of feeding, a single dose of GalN (650 mg/kg) was administered 24 h before their sacrifice. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and tumor necrosis factor-alpha (TNF-alpha) levels were significantly increased after an injection with GalN (P<0.05), but pinitol supplementation at the level of 0.5% reversed these changes to normal levels. Significant decreases in serum triglyceride and cholesterol and increases in hepatic cholesterol were observed in GalN-intoxicated rats. However, supplementation with pinitol significantly attenuated these trends. In addition, pinitol elevated the Mn-superoxide dismutase, glutathione reductase, and catalase activities, prevented hepatic lipid peroxidation, and restored the hepatic GSH levels and cytochrome P450 2E1 function. Thus, 0.5% pinitol supplementation protected the rats from the hepatotoxicity induced by GalN, at least part of its effect being attributable to attenuation of the oxidative stress and inflammatory process promoted by GalN.

Bamboo culm extract supplementation elevates HDL-cholesterol and ameliorates oxidative stress in C57BL/6 mice fed atherogenic diet.

Previously, we reported that bamboo culms possess a stronger antioxidative capacity than bamboo leaves in vitro. In this study, we investigated whether bamboo culm extract (BCE) supplementation ameliorates oxidative stress and hepatic nuclear factor kappaB (NF kappa B) activation in C57BL/6 mice fed an atherogenic diet. In addition, the effect of BCE supplementation on plasma lipid levels of the animals was tested. The mice were randomly assigned to a normal diet, an atherogenic diet (control), or an atherogenic diet supplemented with 1% (wt/wt) BCE or 3% (wt/wt) BCE for 16 weeks. Atherogenic diet-induced oxidative stress, measured by hepatic thiobarbituric acid-reactive substances and protein carbonyls, was significantly lower in the BCE-supplemented groups than in the control (P < .05). Total antioxidative capacity was elevated in the BCE groups, along with greater activities of antioxidative enzymes such as superoxide dismutase and catalase, compared to the control or normal groups (P < .05). The hepatic NF kappa B binding activities were significantly lower in the BCE groups as well (P < .05). The high-density lipoprotein-cholesterol level was significantly elevated by BCE supplementation (P < .05), whereas the effects of BCE on triglyceride and total cholesterol were inconsistent. Results from this study suggest that BCE supplementation may lessen oxidative stress via a series of changes, including a reinforced antioxidant system, and also suggest that the lowered oxidative stress status may down-regulate the activation of inflammatory mediators.

Romidepsin (depsipeptide) induced cell cycle arrest, apoptosis and histone hyperacetylation in lung carcinoma cells (A549) are associated with increase in p21 and hypophosphorylated retinoblastoma proteins expression.

Histone deacetylase inhibitor such as romidepsin (depsipeptide, FR901228, FK228) is a promising new class of antineoplastic agent with the capacity to induce growth arrest and/or apoptosis of cancer cells. However, their precise mechanism of action is uncertain. Histone acetylation and deacetylation are involved in transcriptional activation and transcriptional repression, respectively. Romidepsin induced histone hyperacetylation can be correlated with the cell cycle arrest and apoptosis. In the present study, we investigated the effects of romidepsin on cell proliferation, cell cycle arrest, apoptosis and histone hyperacetylation. Expression of Cdc2/Cdk-1, cyclin B1, cyclin A, p21/Cip1, pRb, pRb2/p130, histone H4 and H3 acetylation status were studied with western blot analysis. The induction of apoptosis has been demonstrated by annexin V-FITC binding assay. Extent of apoptosis has been assessed measuring the activity of caspase-3. Romidepsin led to substantial decrease in the expression of Cdc2/Cdk-1, cyclin B1 and phosphorylated pRb and increase in p21. The pRb protein was found to be one of the targets for the romidepsin induced cell cycle arrest. Flow cytometric analysis showed that romidepsin induced cell cycle arrest at G2-M transition, with significant induction of apoptosis at 25 and 50 nM concentration of romidepsin, with an increase in the number of both early and late apoptotic cells. From this study it is concluded that romidepsin inhibit advanced human lung carcinoma (A549) cell proliferation by altering the expression of cell cycle regulators and apoptotic protein.

Depsipeptide a histone deacetlyase inhibitor down regulates levels of matrix metalloproteinases 2 and 9 mRNA and protein expressions in lung cancer cells (A549).

BACKGROUND: A novel histone deacetylase inhibitor, depsipeptide FR901228 (FK228), is a promising anticancer and antiproliferative agent and has been proposed to regulate gene transcription and reported to lower the risk of several cancers in different cell lines. Depsipeptide showed therapeutic efficacy in Phase I trial of patients with malignant lymphoma. Based on the recognition that basement membrane disruption occurs in acute lung injury and that matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) can degrade type IV collagen, one of the major components of the basement membrane and known to involve in tumor invasion and metastases. We hypothesized that depsipeptide would modulate MMP-2 and MMP-9 production in lung adenocarcinoma cells line (A549). METHODS: In this study, we observed the precise involvement of depsipeptide role on cancer metastasis. A549 cells were treated with depsipeptide at various concentrations (50 and 100nm), for 24h period and then subjected to mRNA levels with RT-PCR and protein levels with Western blot analysis to investigate the impact of depsipeptide on MMP-2 and MMP-9 expressions and further confirmed by using immunocytochemistry. RESULTS: The results showed that depsipeptide treatment decreased the expressions of MMP-2 and MMP-9 in dose-dependent manner. The level of mRNA and proteins expressions were significantly decreased in depsipeptide treated A549 cells in a dose-dependent manner and the level of pro-MMP-9 was found to be high in the 100nm depsipeptide-treated cell lysate of A549 cells, suggesting inhibitory role of depsipeptide on pro-MMP-9 activation. Further immunocytochemistry studies showed the weak expression of MMP-2 and MMP-9 in depsipeptide treated cells. CONCLUSION: We speculate that inhibition of metastasis-specific MMPs in cancer cells may be one of the targets for anticancer function of depsipeptide, and thus provides the molecular basis for the development of depsipeptide as a novel chemopreventive agent for metastatic lung cancer.

Cloning and functional expression of the gene encoding an inhibitor against Aspergillus flavus alpha-amylase, a novel seed lectin from Lablab purpureus (Dolichos lablab).

Maize is one of the more important agricultural crops in the world and, under certain conditions, prone to attack from pathogenic fungi. One of these, Aspergillus flavus, produces toxic and carcinogenic metabolites, called aflatoxins, as byproducts of its infection of maize kernels. The alpha-amylase of A. flavus is known to promote aflatoxin production in the endosperm of these infected kernels, and a 36-kDa protein from the Lablab purpureus, denoted AILP, has been shown to inhibit alpha-amylase production and the growth of A. flavus. Here, we report the isolation of six full-length labAI genes encoding AILP and a detailed analysis of the activities of the encoded proteins. Each of the six labAI genes encoded sequences of 274 amino acids, with the deduced amino acid sequences showing approximately 95-99% identity. The sequences are similar to those of lectin members of a legume lectin-arcelin-alpha-amylase inhibitor family reported to function in plant resistance to insect pests. The labAI genes did not show any of the structures characteristic of conserved structures identified in alpha-amylase inhibitors to date. The recombinant proteins of labAI-1 and labAI-2 agglutinated human red blood cells and inhibited A. flavus alpha-amylase in a manner similar to that shown by AILP. These data indicate that labAI genes are a new class of lectin members in legume seeds and that their proteins have both lectin and alpha-amylase inhibitor activity. These results are a valuable contribution to our knowledge of plant-pathogen interactions and will be applicable for developing protocols aimed at controlling A. flavus infection.

[The attitudes of nurses toward transsexuals].

PURPOSE: The purpose of this study was to identify the attitude of nurses toward transsexuals. METHOD: The Q-methodology which provides a method of analyzing the subjectivity of each item was used. Twenty-nine nurses classified the 50 selected Q-statements into a normal distribution using a 9 point scale. The collected data was analyzed using the Quanl PC program. RESULT: Four types of attitudes toward transsexuals were identified. The first type (humanitarian acceptance) showed an attitude of respecting transsexuals as human beings and understanding and accepting their desires and difficulties. The second type (superficial understanding) understood the psychological conflicts and suffering of transsexuals but could not accept them as members of families or society. The third type (insufficient understanding) did not feel a sense of rejection toward transsexuals but showed a lack of understanding of their desires and difficulties. The fourth type (rejection) failed to understand the desires and difficulties of transsexuals and showed a sense of rejection toward them, in addition to regarding them as sexually immoral people. CONCLUSION: The results of the study indicate that different approaches of educational programs based on the four types of attitudes toward transsexuals are recommended.