Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

Modulations of the Bcl-2/Bax family were involved in the chemopreventive effects of licorice root (Glycyrrhiza uralensis Fisch) in MCF-7 human breast cancer cell

Recently, cancer chemoprevention with strategies using foods and medicinal herbs has been regarded as one of the most visible fields for cancer control. Genistein in soy, American ginseng, and resveratrol are well-known to have antiproliferative properties in human breast cancer. Licorice root is a botanical, a shrub native to southern Europe and Asia, which primarily has desirable qualities in sweetening and herbal medicine. In this study, licorice (Glycyrrhiza uralensis Fisch) root also inhibits cell proliferation in human breast cancer cell. The cell proliferation study demonstrated that licorice root reduced the proliferation of MCF-7 cells in a dose- and time-dependent manner. The extracts were fractionated in CHCl(3), EtOAc, C(6)H(14), and CH(3)OH-H(2)O (70:30), and these extracts of licorice root (50 microg/mL) induced DNA fragmentation demonstrated by Hoechst 33258 staining. Apoptosis also determined the sub-G1 accumulation by flow cytometry analysis. These results were consistent with specific cleavage of PARP and antiapoptotic protein Bcl-2 and up-regulation of proapoptotic protein Bax demonstrated by Western blotting. Our findings suggest that licorice root may have chemopreventive effects against human breast cancer through the modulation of the expression of the Bcl-2/Bax family of apoptotic regulatory factors.     

Ovicidal and adulticidal activity of Eucalyptus globulus leaf oil terpenoids against Pediculus humanus capitis (Anoplura: Pediculidae)

The toxic effects of Eucalyptus globulus leaf oil-derived monoterpenoids [1,8-cineole, l-phellandrene, (-)-alpha-pinene, 2-beta-pinene, trans-pinocarveol, gamma-terpinene, and 1-alpha-terpineol] and the known Eucalyptusleaf oil terpenoids (beta-eudesmol and geranyl acetate) on eggs and females of the human head louse, Pediculus humanus capitis, were examined using direct contact and fumigation bioassays and compared with the lethal activity of delta-phenothrin and pyrethrum, two commonly used pediculicides. In a filter paper contact bioassay with female P. h. capitis, the pediculicidal activity was more pronounced with Eucalyptus leaf oil than with either delta-phenothrin or pyrethrum on the basis of LT(50) values (0.125 vs 0.25 mg/cm(2)). 1,8-Cineole was 2.2- and 2.3-fold more toxic than either delta-phenothrin or pyrethrum, respectively. The pediculicidal activities of (-)-alpha-pinene, 2-beta-pinene, and (E)-pinocarveol were comparable to those of delta-phenothrin and pyrethrum. l-Phellandrene, gamma-terpinene, and 1-alpha-terpineol were relatively less active than delta-phenothrin and pyrethrum. beta-Eudesmol and geranyl acetate were ineffective. 1-alpha-Terpineol and (E)-pinocaveol were highly effective at 0.5 and 1.0 mg/cm(2), respectively, against P. h. capitis eggs. At 1.0 mg/cm(2), (-)-alpha-pinene, 2-beta-pinene, and gamma-terpinene exhibited moderate ovicidal activity, whereas little or no ovicidal activity was observed with the other terpenoids and with delta-phenothrin and pyrethrum. In fumigation tests with female P. h. capitis at 0.25 mg/cm(2), 1,8-cineole, (-)-alpha-pinene, (E)-pinocarveol, and 1-alpha-terpineol were more effective in closed cups than in open ones, indicating that the effect of the monoterpenoids was largely due to action in the vapor phase. Neither delta-phenothrin nor pyrethrum exhibited fumigant toxicity. Eucalyptus leaf oil, particularly 1,8-cineole, 1-alpha-terpineol, and (E)-pinocaveol, merits further study as potential pediculicides or lead compounds for the control of P. h. capitis.     

His-His-Leu, an angiotensin I converting enzyme inhibitory peptide derived from Korean soybean paste, exerts antihypertensive activity in vivo.

It has been reported that soybean peptide fractions isolated from Korean fermented soybean paste exert angiotensin I converting enzyme (ACE) inhibitory activity in vitro. In this study, further purification and identification of the most active fraction inhibiting ACE activity were performed, and its antihypertensive activity in vivo was confirmed. Subsequently, a novel ACE inhibitory peptide was isolated by preparative HPLC. The amino acid sequence of the isolated peptide was identified as His-His-Leu (HHL) by Edman degradation. The IC(50) value of the HHL for ACE activity was 2.2 microg/mL in vitro. Moreover, the synthetic tripeptide HHL (spHHL) resulted in a significant decrease of ACE activity in the aorta and led to lowered systolic blood pressure (SBP) in spontaneously hypertensive (SH) rats compared to control. Triple injections of spHHL, 5 mg/kg of body weight/injection resulted in a significant decrease of SBP by 61 mmHg (p < 0.01) after the third injection. These results demonstrated that the ACE inhibitory peptide HHL derived from Korean fermented soybean paste exerted antihypertensive activity in vivo.     

Growth-inhibiting effects of Coptis japonica root-derived isoquinoline alkaloids on human intestinal bacteria.

The growth-inhibiting activity of Coptis japonica (Makino) root-derived materials toward eight human intestinal bacteria was examined using an impregnated paper disk method and compared to that of four commercially available isoquinoline alkaloids [berberine sulfate (BS), berberine iodide (BI), palmatine chloride (PC), and palmatine sulfate(PS)], as well as that of Thea sinensis leaf-derived epigallocatechin gallate (EGCG). The biologically active constituents of the Coptis extract were characterized as the isoquinoline alkaloids berberine chloride (BC), palmatine iodide (PI), and coptisine chloride (CC) by spectral analysis. The growth responses varied with both chemical and bacterial strain used. In a test using 500 microg/disk, BC and PI produced a clear inhibitory effect against Bifidobacterium longum, Bifidobacterium bifidum, Clostridium perfringens, and Clostridium paraputrificum, whereas weak or no inhibition was observed in Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, and Escherichia coli. At 1000 microg/ disk, CC revealed weak or no growth inhibition toward all test bacteria, whereas EGCG exhibited weak growth inhibition against only C. perfringens and C. paraputrificum. Among various isoquinoline alkaloids, BC exhibited more potent inhibitory activity toward C. perfringens than BI and BS, whereas the inhibitory effect was more pronounced in PI compared to PC and PS. The Coptis root-derived materials did not promote growth of B. longum and C. perfringens.     

Effect of far-infrared radiation on the antioxidant activity of rice hulls

After far-infrared (FIR) radiation onto rice hull, a methanolic extract was prepared for the determination of antioxidant ability. After 30 min of FIR treatment, the radical scavenging activity and total phenol contents of rice hull extracts increased from 47.74 to 79.63% and from 0.12 to 0.19 mM, respectively, compared to control. Inhibition of lipid peroxidation in extracts was also increased from 41.07 to 47.96%. According to the GC-MS analysis, more phenolic compounds (p-coumaric acid, 3-vinyl-1-oxybenzene, p-hydroxybenzaldehyde, vanillin, p-hydroxybenzoic acid, and 4,7-dihydroxyvanillic acid) were detected in FIR-irradiated rice hull extract. These results indicated that FIR radiation onto rice hull could liberate and activate covalently bound phenolic compounds that have antioxidant activities.     

Panax ginseng extract rich in ginsenoside protopanaxatriol attenuates blood pressure elevation in spontaneously hypertensive rats by affecting the Akt-dependent phosphorylation of endothelial nitric oxide synthase

Nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS) is a fundamental regulator of systemic blood pressure. Ginsenosides from Panax ginseng have been investigated in vitro for the molecular and biochemical mechanisms by which they stimulate NO release in vascular endothelial cells; however, little research has been done to confirm the physiological relevance of these in vitro studies. To address this research gap, the effects of a P. ginseng extract rich in ginsenosides from protopanaxatriol on spontaneously hypertensive rats (SHRs) was examined. Ginseng extract administration stimulated nongenomic Akt-mediated eNOS activation, enhanced NO production, improved vessel wall thickening, and alleviated hypertension in SHRs, confirming the physiological relevance of previous in vitro studies with ginsenosides.     

Selective growth-inhibiting effects of compounds identified in Tabebuia impetiginosa inner bark on human intestinal bacteria

The growth-inhibiting activity of anthraquinone-2-carboxylic acid and lapachol identified in the inner bark of taheebo, Tabebuia impetiginosa, toward 10 human intestinal bacteria was evaluated by using a paper disk diffusion bioassay and compared to those of seven lapachol congeners (1,4-naphthoquinone, naphthazarin, menadione, lawsone, plumbagin, juglone, and dichlone) as well as two commercially available antibiotics, chloramphenicol and tetracycline. Anthraquinone-2-carboxylic acid exhibited very strong growth inhibition of Clostridium paraputrificum at 1 microg/disk while 100 microg/disk of lapachol was needed for moderate growth inhibition of the same organism. These two isolates exhibited weak inhibition of Clostridium perfringens and Escherichia coli at 100 microg/disk while no adverse effects were observed on the growth of Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium infantis, Lactobacillus acidophilus, and Lactobacillus casei at 1000 microg/disk. Structure-activity relationships indicate that a methyl group in the C-2 position of 1,4-naphthoquinone derivatives might play an important role in antibacterial activity.     

Identification of a RAPD marker linked to a brown planthopper resistance gene in rice

We report the tagging of a brown planthopper (BPH) resistance gene (Bph–1) in rice using RAPD and RFLP markers. The Korean rice variety ‘Gayabyeo’ has dominant duplicate genes including Bph–1 conferring resistance to biotype 1 of BPH. Bulked segregant RAPD analysis was employed for rapid identification of DNA markers linked to resistance genes. For tagging these two genes, an F2F3 population from a ‘Gayabyeo’ × ‘Nagdongbyeo’ cross was developed and evaluated for BPH resistance. Three bulked DNAs from two groups of homozygous BPH resistant (each for Bph–1 and the other unknown gene) and homozygous susceptible F2 plants were analyzed by RAPD using 140 random oligomers. One primer, OPD–7 yielded a 700-bp fragment that was present in Gayabyeo and resistant F2 plants (homozygous for Bph-1 locus) but absent in Nagdongbyeo and susceptible F2 plants. Cosegregation of this marker with Bph-1 was verified using an F2 population segregating for Bph-1. Chromosomal regions surrounding the Bph-1 were examined with additional RFLP and microsatellite markers on chromosome 12 to define the location of the RAPD marker and Bph-1. Use of this RAPD marker could facilitate early selection of resistant lines for BPH. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nutritional Properties of Various Tissues from New Kenaf Cultivars

Kenaf is an economically important crop that contains various functional compounds. The objective of this study was to evaluate the nutritional properties of leaves, stem bark, flowers, and seeds collected from three mutant kenaf cultivars (Jangdae, Baekma, and Jeokbong) and two original cultivars (Jinju and C14). For all analyzed tissues, there were no significant differences in the proximate compositions (moisture, crude protein, crude fiber, crude fat, ash, and nitrogen-free extract) among the tested genotypes, except for the leaf crude protein content, which was highest in Jinju plants. Of the tested minerals, potassium and calcium were the most abundant in all tissues. Additionally, we detected significant differences in the mineral contents (e.g., calcium, phosphorus, potassium, magnesium, zinc, and iron) of various tissues in all cultivars. Proline and phenylalanine were the major amino acids detected in leaves, and the highest total amino acid (TAA) and essential amino acid (EAA) contents were observed in Jinju plants. The TAA and EAA contents were lower in stem bark than in the other tissues, except in Baekma plants, in which the lowest levels were observed in flowers. Moreover, the TAA and EAA contents were approximately 20 times lower in white flowers (Baekma) than in ivory flowers. Furthermore, the highest TAA and EAA concentrations were observed in Jangdae seeds. These results may be useful for identifying the optimal cultivar and tissues for use in food products.