Objective: To evaluate effects of epigallocatechin-3-gallate (EGCG) on the viability, membrane properties, and zinc distribution, with and without the presence of Zn²⁺, in human prostate carcinoma LNCaP cells. Methods: We examined changes in cellular morphology and membrane fluidity of LNCaP cells, distribution of cellular zinc, and the incorporated portion of EGCG after treatments with EGCG, Zn²⁺, and EGCG+Zn²⁺. Results: We observed an alteration in cellular morphology and a decrease in membrane fluidity of LNCaP cells after treatment with EGCG or Zn²⁺. The proportion of EGCG incorporated into liposomes treated with the mixture of EGCG and Zn²⁺ at the ratio of 1:1 was 90.57%, which was significantly higher than that treated with EGCG alone (30.33%). Electron spin resonance (ESR) studies and determination of fatty acids showed that the effects of EGCG on the membrane fluidity of LNCaP were decreased by Zn²⁺. EGCG accelerated the accumulation of zinc in the mitochondria and cytosol as observed by atomic absorption spectrometer. Conclusion: These results show that EGCG interacted with cell membrane, decreased the membrane fluidity of LNCaP cells, and accelerated zinc accumulation in the mitochondria and cytosol, which could be the mechanism by which EGCG inhibits proliferation of LNCaP cells. In addition, high concentrations of Zn²⁺ could attenuate the actions elicited by EGCG.

The sprayed chemicals on tea leaves disappear over a period of time by the influence of rainfall elution, evaporation, growth dilution, and photodegradation. Influence of plant growth on the four fungicides (hexaconazole, propiconazole, tridemorph, and c) was studied to know the constructive loss of fungicides. The study shows that residues of fungicides sprayed on tea shoots got diluted by the growing process. The expansion of a leaf took 8 to 11 d and more than 50% of the fungicide residues were cleaned out during this leaf expansion period. Under south Indian agroclimatic condition, the fungicides are sprayed at an interval of 10 d, so it is safe that the tea is harvested on the 10th day of the application of fungicides.

Low temperature stress during germination and early seedling growth is an important constraint of global production of maize. The effects of seed priming with 0.25%, 0.50%, and 0.75% (w/v) chitosan solutions at 15 °C on the growth and physiological changes were investigated using two maize (Zea mays L.) inbred lines, HuangC (chilling-tolerant) and Mo17 (chilling-sensitive). While seed priming with chitosan had no significant effect on germination percentage under low temperature stress, it enhanced germination index, reduced the mean germination time (MGT), and increased shoot height, root length, and shoot and root dry weights in both maize lines. The decline of malondialdehyde (MDA) content and relative permeability of the plasma membrane and the increase of the concentrations of soluble sugars and proline, peroxidase (POD) activity, and catalase (CAT) activity were detected both in the chilling-sensitive and chilling-tolerant maize seedlings after priming with the three concentrations of chitosan. HuangC was less sensitive to responding to different concentrations of chitosan. Priming with 0.50% chitosan for about 60~64 h seemed to have the best effects. Thus, it suggests that seed priming with chitosan may improve the speed of germination of maize seed and benefit for seedling growth under low temperature stress.

Fasciclin family proteins have been identified as cell adhesion molecules in various organisms. In this study, a novel Magnaporthe oryzae fasciclin-like protein encoding gene, named MoFLP1, was isolated from a subtractive suppressive cDNA library and functionally analyzed. Sequence analysis showed that the MoFLP1 gene contains an open reading frame (ORF) of 1050 nucleotides encoding 349 amino acids with a calculated molecular weight of 35.85 kDa and a pI of 7.76. The deduced MoFLP1 protein contains a 17-amino acid secretion signal sequence and an 18-amino acid sequence with the characteristics of a glycosylphosphotidylinositol (GPI) anchor additional signal at its N- and C-terminuses, respectively. Potential N-glycosylation sites and domains involving cell adhesion were also identified in MoFLP1. Sequence analysis and subcellular localization by the expression of MoFLP1-GFP fusion construct in M. oryzae indicated that the MoFLP1 protein is probably localized on the vacuole membrane. Two MoFLP1 null mutants generated by targeted gene disruption exhibited marked reduction of conidiation, conidial adhesion, appressorium turgor, and pathogenicity. Our results indicate that fasciclin proteins play important roles in fungal development and pathogenicity in M. oryzae.

Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune response to parasitoid attack, we explored the response of Papilio xuthus parasitized by the endoparasitic wasp Pteromalus puparum using proteomic approach. By examining the differential expression of plasma proteins in the parasitized and unparasitized host pupae by two-dimensional (2D) electrophoresis, 16 proteins were found to vary in relation to parasitization compared with unparasitized control samples. All of them were submitted to identification by mass spectrometry coupled with a database search. The modulated proteins were found to fall into the following functional groups: humoral or cellular immunity, detoxification, energy metabolism, and others. This study contributes insights into the molecular mechanism of the relationships between parasitoids and their host insects.

The effects of CO₂ enrichment on the growth and glucosinolate (GS) concentrations in the bolting stem of Chinese kale (Brassica alboglabra L.) treated with three nitrogen (N) concentrations (5, 10, and 20 mmol/L) were investigated. Height, stem thickness, and dry weights of the total aerial parts, bolting stems, and roots, as well as the root to shoot ratio, significantly increased as CO₂ concentration was elevated from 350 to 800 μl/L at each N concentration. In the edible part of the bolting stem, 11 individual GSs were identified, including 7 aliphatic and 4 indolyl GSs. GS concentration was affected by the elevated CO₂ concentration, N concentration, and CO₂×N interaction. At 5 and 10 mmol N/L, the concentrations of aliphatic GSs and total GSs significantly increased, whereas those of indolyl GSs were not affected, by elevated atmospheric CO₂. However, at 20 mmol N/L, elevated CO₂ had no significant effects on the concentrations of total GSs and total indolyl GSs, but the concentrations of total aliphatic GSs significantly increased. Moreover, the bolting stem carbon (C) content increased, whereas the N and sulfur (S) contents decreased under elevated CO₂ concentration in the three N treatments, resulting in changes in the C/N and N/S ratios. Also the C/N ratio is not a reliable predictor of change of GS concentration, while the changes in N and S contents and the N/S ratio at the elevated CO₂ concentration may influence the GS concentration in Chinese kale bolting stems. The results demonstrate that high nitrogen supply is beneficial for the growth of Chinese kale, but not for the GS concentration in bolting stems, under elevated CO₂ condition.

Near-infrared (NIR) transmittance spectroscopy combined with least-squares support vector machine (LS-SVM) was investigated to study the quality change of tomato juice during the storage. A total of 100 tomato juice samples were used. The spectrum of each tomato juice was collected twice: the first measurement was taken when the tomato juice was fresh and had not undergone any changes, and the second measurement was taken after a month. Principal component analysis (PCA) was used to examine a potential capability of separating juice before and after the storage. The soluble solid content (SSC) and pH of the juice samples were determined. The results show that changes in certain compounds between tomato juice before and after the storage period were obvious. An excellent precision was achieved by LS-SVM model compared with discriminant partial least-squares (DPLS), soft independent modeling of class analogy (SIMCA), and discriminant analysis (DA) models, with 100% of a total accuracy. It can be found that NIR spectroscopy coupled with LS-SVM, DPLS, SIMCA, and DA can be used to control the quality change of tomato juice during the storage.

In this paper, we presented a novel method for the facile and efficient one-pot synthesis of 2-arylbenzoxazoles, which were directly synthesized from 2-aminophenol and aldehydes catalyzed by hydrogen tetrachloroaurate (HAuCl₄·4H₂O) under an oxygen atmosphere with anhydrous tetrahydrofuran (THF) as solvent or in solvent-free condition. The results show that this method could bring excellent yields as high as 96%. THF was proven to be the best choice among several solvents screened and the reaction was tolerated with a variety of aromatic aldehydes possessing electron-donating or withdrawing groups. The advantages of the present method lie in catalytic process using economic and environmentally benign dioxygen as oxidant.

Site-directed mutagenesis (SDM) has been a very important method to probe the function-structure relationship of proteins. In this study, we introduced an easy-to-use, polymerase chain reaction (PCR)-based SDM method for double-stranded plasmid DNA, with a designed restriction site to ensure simple and efficient mutant screening. The DNA sequence to be mutated was first translated into amino acid sequence and then the amino acid sequence was reversely translated into DNA sequence with degenerate codons, resulting in a large number of sequences with silent mutations, which contained various restriction endonuclease (RE) sites. Certain mutated sequence with an appropriate RE site was selected as the target DNA sequence for designing a pair of mutation primers to amplify the full-length plasmid via inverse PCR. The amplified product was 5′-phosphorylated, circularized, and transformed into an Escherichia coli host. The transformants were screened by digesting with the designed RE. This protocol uses only one pair of primers and only one PCR is conducted, without the need for hybridization with hazardous isotope for mutant screening or subcloning step.