Effects of physical states of phospholipids on the incorporation and cytochalasin B binding activity of human erythrocyte membrane proteins in reconstituted vesicles

Proteoliposomes were reconstituted from a Triton extract of human erythrocyte membrane proteins and a mixture of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of varying ratios. With mixtures of egg PC and soybean PE, the protein/lipid ratio of the reconstituted vesicles was maximal at 25% PC and 75% PE, the composition which is known to have a maximum bilayer disruption (highest occurrence of lipidic particles seen by freeze-fracture electron microscopy). With mixtures of 1-palmitoyl-2-oleoyl-PC and dilinoleoyl-PE, which gave vesicles with few isolated lipidic particles at room temperature, the effect was less pronounced. The specific activity of the cytochalasin B (CB) binding protein in the reconstituted vesicles, on the other hand, was increased monotonically up to severalfold as the PC content was increased in the egg PC/soybean PE mixture. A similar increase was observed when soybean PE was partially substituted by dimyristoyl-PC, cholesterol, or transphosphatidylated PE from egg PC. These findings indicate that preexisting defects in the lipid bilayer promote protein incorporation into the bilayer during reconstitution whereas reduction of the bilayer fluidity facilitates the CB binding activity in the reconstituted vesicles.     

Detection of beta-Glucosidase Activity in Polyacrylamide Gels with Esculin as Substrate

beta-Glucosidase can be located after nondenaturing polyacrylamide gel electrophoresis by incubating the gel with 0.1% esculin and 0.03% ferric chloride. The esculetin released from esculin by beta-glucosidase action reacts with ferric ion to produce a black band, corresponding to the beta-glucosidase, against the transparent background.     

Feeding response of the cotton aphid,Aphis gossypii,to sublethal rates of flonicamid and imidacloprid

The cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is an important sap‐sucking pest of many plants, including melons and peppers. This study was conducted to determine the effects of sublethal exposure to flonicamid and imidacloprid and the mechanisms by which these insecticides affect the feeding behavior of A. gossypii. The median lethal concentrations (LC₅₀) of flonicamid and imidacloprid for adult A. gossypii were 2.40 and 1.92 mg l^?1, respectively. The lower lethal concentrations of flonicamid were 1.01 mg l^?1 (LC₃₀) and 0.29 mg l^?1 (LC₁₀), and those of imidacloprid were 0.82 mg l^?1 (LC₃₀) and 0.24 mg l^?1 (LC₁₀). The developmental period of A. gossypii nymphs at LC₃₀ was 3.6 days for both insecticides, which was shorter than that of the untreated controls (4.2 days). Longevity and total fecundity of A. gossypii adults were decreased at the sublethal concentrations of both insecticides. The lowest net reproductive rate was observed in A. gossypii treated with the LC₃₀ of flonicamid. Feeding behavior analyses using an electrical penetration graph showed that sublethal concentrations of flonicamid and imidacloprid had significant effects on the duration of phloem ingestion. Higher doses of flonicamid induced starvation by inhibiting phloem ingestion, whereas imidacloprid acted as a contact toxin rather than an inhibitor of feeding behavior.     

Evaluation of the acaricidal activity of 63 commercialized pesticides against Haemaphysalis longicornis (Acari: Ixodidae)

The acaricidal activity and residual effects of 63 commercialized pesticides against the Cheongju strain of Haemaphysalis longicornis in Korea were examined. Twenty‐two pesticides (4 carbamates, 5 organophosphates, 10 pyrethroids, 1 amitraz, 1 diamide, and 1 unclassified pesticide) caused greater than 80% mortality in H. longicornis adults under laboratory conditions using the spray method. These 22 pesticides were used to treat grassland under field conditions for investigation of the residual effects, and 100% mortality in H. longicornis adults was observed with all the carbamates (carbosulfan, benfuracarb, fenobucarb, and carbaryl) and γ‐cyhalothrin after 3 days of pesticide treatment. γ‐Cyhalothrin exhibited 56.7% mortality after 10 days of treatment, which was the longest residual effect of treated grass on H. longicornis adults. With regard to the residual effects on H. longicornis under field conditions, most of the 22 pesticides exhibited higher mortality in nymphs than in adults. In particular, benfuracarb exhibited 96.7% acaricidal activity until 10 days after treatment. These results indicate that carbamates are highly likely to be available under field conditions and, based on this preliminary data, could be used for the control of H. longicornis adults and nymphs.     

Synthesis and biological evaluation of 1-(6-methylpyridin-2-yl)-5-(quinoxalin-6-yl)-1,2,3-triazoles as transforming growth factor-β type 1 receptor kinase inhibitors

A series of 1-(6-methylpyridin-2-yl)-5-(quinoxalin-6-yl)-1,2,3-triazoles has been synthesized and evaluated for their ALK5 inhibitory activity. The 1-(6-methylpyridin-2-yl)-1,2,3-triazoles were assembled by Cu(I)-catalyzed azide–alkyne 1,3-dipolar cycloaddition. Following this, quinoxaline was introduced through Pd-catalyzed direct arylation. The synthesized 1-(6-methylpyridin-2-yl)-5-(quinoxalin-6-yl)-1,2,3-triazoles revealed significant selectivity differences with respect to p38α MAP kinase. In particular, 12k showed 80.8% ALK5 inhibitory activity at a concentration of 10 μM and IC₅₀ value of 4.69 μM, but did not show p38α MAP kinase inhibitory activity (?1.94% inhibition at a concentration of 10 μM).     

Isolation of Aeromonas salmonicida from masu salmon in the Republic of Korea

The isolation and identification of the Gram-negative fish pathogen, Aeromonas salmonicida salmonicida, from masu salmon (Oncorhynchus masou) at the Yang Yang Salmon Hatchery in the Republic of Korea is described. The bacterium possessed certain characteristics in common with isolates from fish in Japan. It is an important cause of mortality among cultured salmonids in the Republic of Korea. This is the first report of this fish pathogen from the Asian mainland.     

A colorimetric sandwich-type assay for sensitive thrombin detection based on enzyme-linked aptamer assay

A colorimetric sandwich-type assay based on enzyme-linked aptamer assay has been developed for the fast and sensitive detection of as low as 25 fM of thrombin with high linearity. Aptamer-immobilized glass was used to capture the target analyte, whereas a second aptamer, functionalized with horseradish peroxidase (HRP), was employed for the conventional 3,5,3′,5′-tetramethylbenzidine (TMB)-based colorimetric detection. Without the troublesome antibody requirement of the conventional enzyme-linked immunosorbent assay (ELISA), as low as 25 fM of thrombin could be rapidly and reproducibly detected. This assay has superior, or at least equal, recovery and accuracy to that of conventional antibody-based ELISA.     

Structure tuning of pyrazolylpyrrole derivatives as ERK inhibitors utilizing dual tools; 3D-QSAR and side-chain hopping

The ERK pathway is a well-known therapeutic target of cancer treatment with great advantage of selectivity between normal cells and cancer cells, and the number of direct ERK kinase inhibitors is quite limited considering large number of available ERK structure from PDB. Therefore, we tried to combine 3D-QSAR with side-chain hopping in an attempt to produce novel structures as ERK inhibitors. The predictive models with q(2) value of 0.867, r(2) value of 0.991 in CoMFA and q(2) value of 0.628, r(2) value of 0.950 in CoMSIA were used to select effective compounds from new library generated from side-chain hopping by CombiGlide.     

Structure based design and syntheses of amino-1H-pyrazole amide derivatives as selective Raf kinase inhibitors in melanoma cells

The synthesis of a novel series of N-(5-amino-1-(4-methoxybenzyl)-1H-pyrazol-4-yl amide derivatives 6a-o, 7a-s and their antiproliferative activities against A375P melanoma cell line were described. Most compounds showed competitive antiproliferative activities to sorafenib, the reference standard. Among them, N-(5-amino-1-(4-methoxybenzyl)-1H-pyrazol-4-yl)-5-(3-(4-chloro-3-(trifluoromethyl)phenyl) ureido)-2-methylbenzamide 7c exhibited potent activities (GI(50)=0.27 μM). Especially, 7c was found to be a potent and selective B-Raf V600E and C-Raf inhibitor (IC(50)=0.26 μM, IC(50)=0.11 μM, respectively), showing a possibility as melanoma therapeutics.