Antiproliferative and Antimicrobial Potentials of a Lectin from Aplysia kurodai (Sea Hare) Eggs

In recent years, there has been considerable interest in lectins from marine invertebrates. In this study, the biological activities of a lectin protein isolated from the eggs of Sea hare (Aplysia kurodai) were evaluated. The 40 kDa Aplysia kurodai egg lectin (or AKL-40) binds to D-galacturonic acid and D-galactose sugars similar to previously purified isotypes with various molecular weights (32/30 and 16 kDa). The N-terminal sequence of AKL-40 was similar to other sea hare egg lectins. The lectin was shown to be moderately toxic to brine shrimp nauplii, with an LC₅₀ value of 63.63 µg/mL. It agglutinated Ehrlich ascites carcinoma cells and reduced their growth, up to 58.3% in vivo when injected into Swiss albino mice at a rate of 2 mg/kg/day. The morphology of these cells apparently changed due to AKL-40, while the expression of apoptosis-related genes (p53, Bax, and Bcl-XL) suggested a possible apoptotic pathway of cell death. AKL-40 also inhibited the growth of human erythroleukemia cells, probably via activating the MAPK/ERK pathway, but did not affect human B-lymphoma cells (Raji) or rat basophilic leukemia cells (RBL-1). In vitro, lectin suppressed the growth of Ehrlich ascites carcinoma and U937 cells by 37.9% and 31.8%, respectively. Along with strong antifungal activity against Talaromyces verruculosus, AKL showed antibacterial activity against Staphylococcus aureus, Shigella sonnei, and Bacillus cereus whereas the growth of Escherichia coli was not affected by the lectin. This study explores the antiproliferative and antimicrobial potentials of AKL as well as its involvement in embryo defense of sea hare.     

Fusariosis in rubber tree: pathogenic,morphological, and molecular characterization of the causal agent

Fusariosis, one of the phytosanitary problems found in rubber producing areas in the northwest of the state of São Paulo, is a disease that affects the bark of the adult plants, affecting the exploitation of latex. The typical symptoms appear as cracks in the bark that expand from the rootstock towards the tapping panel, causing a drying of the latex flux in the injured region, impeding latex tapping. Due to the recent incidence of this disease in rubber plantations, the goal of this study was to characterize the Fusarium associated with symptomatic rubber tissue in three different locations in the state of São Paulo. In order to identify Fusarium species, pathogenicity, morphological, cultural and molecular studies were carried out. A total of 51 isolates were obtained and separated into three groups based on macroconidium morphology, presence or absence of sporodochia, types of chlamydospores, formation of phialides and conidiogenesis of microconidia and mesoconidia, mycelial growth rate and coloring of the colonies. These groups were corroborated using DNA sequence information for five different genetic loci, and were subsequently recognized as Fusarium oxysporum, F. incarnatum and F. decemcellulare. Our results further showed that all 51 of the Fusarium isolates recovered were pathogenic to rubber tree seedlings of RRIM 600 standard clone. This study also reports for the presence of F. oxysporum and F. incarnatum in rubber plantations in the state of São Paulo and in Brazil.

     

Identification of fructo- and malto-oligosaccharides in cured tobacco leaves (Nicotiana tabacum)

Fructo-oligosaccharides (FOSs) and malto-oligosaccharides (MOSs) in cured tobacco leaves ( Nicotiana tabacum ) were detected and quantified using high-performance liquid chromatography (HPLC) coupled with electrospray ionization (ESI) tandem mass spectrometry (MS/MS). These oligosaccharides were present in several tobacco materials including flue-cured tobacco, sun/air-cured tobacco, and cut filler of commercially available tobacco products, but were not detected in air-cured tobacco. The changes in these oligosaccharides during storage were also investigated. The results revealed that MOSs simply decreased under a warm atmosphere and FOSs increased. In addition, the formation of FOSs in cured tobacco leaves occurred in the presence of sucrose. With regard to FOSs, it has been reported that green tobacco leaves do not contain FOSs such as kestose, nystose, and fructosyl-nystose. The results of a model test suggested that the changes in FOS amount were caused by enzymatic reactions.     

The preventive effect of Bacillus subtilus strain DB9011 against experimental infection with enterotoxcemic Escherichia coli in weaning piglets

Porcine edema disease (ED) is caused by Shiga toxin 2e‐producing Escherichia coli (STEC). Post‐weaned piglets often suffer from ED as a result of intestinal infection with STEC, which causes impaired growth performance and high mortality. Antimicrobial therapy is a curative treatment for piglets infected with STEC, but the emergence of antimicrobial‐resistant STEC has become a serious problem for Japanese pig farmers. Therefore, an alternative strategy other than antimicrobial therapy is needed for the prevention or treatment of ED. In this study, we evaluated the effect of oral administration of Bacillus subtilis DB9011 (DB9011) to prevent the experimental infection of STEC in weaning piglets. Eight 21‐day‐old piglets were divided into two groups: STEC challenge with the basal diet, and STEC challenge with DB9011 supplemented diet. The challenge was carried out when the animals were 25, 26 and 27 days old using STEC contained in capsules resistant against gastric digestion. All pigs were euthanized at 36 days of age. DB9011 improved the symptoms of ED and decreased the number of STEC in the ileal digesta and feces. Accordingly, oral administration of DB9011 in weaned piglets prevents ED through the suppression of the growth of STEC in the ileum.     

Effects of meat species on the postprandial thermic effect in rats

We examined animal species differences in the postprandial thermic effect of meat and investigated the postprandial thermic effect of mutton in rats. After intake of experimental diets containing each meat, body temperatures of rats fed mutton or venison were significantly higher than that of rats fed rabbit meat. After intake of experimental diets containing fractionized mutton, the body temperatures of rats fed diets containing lean mutton protein were higher than those of rats fed diets without lean mutton protein. In a two‐dimensional fluorescence difference gel electrophoresis study of brown adipose tissue, it was shown that the intake of mutton up‐regulated the expression of many signaling molecules that are involved in energy metabolism. The postprandial thermic effect of mutton seems to be due not to catecholamine and adrenocorticotropic hormone but to thyroid hormones. The results suggest that intake of lean mutton protein stimulates the secretion of thyroid hormones and facilitates energy metabolism in rats.     

Anti-Thy-1 Antibody-mediated Complement-dependent Cytotoxicity is Regulated by the Distribution of Antigen,Antibody and Membrane Complement Regulatory Proteins in Rats

Some therapeutic antibodies as anticancer agents exert their effects through the host immune system, but the factors that predict their cytotoxicity, including complement-dependent cytotoxicity (CDC), are unclear. In the present study, we attempted to elucidate some of these factors in a preclinical model. CDC-related mesangiolysis caused by administration of the anti-Thy-1.1 antibody can be studied in the rat anti-Thy-1 glomerulonephritis model, so the model was used in this study. Three animals each were sacrificed at 0.5, 1, 8, 24 and 48 hours after i.v. administration of the anti-Thy-1.1 antibody at 1mg/kg. The distribution of the Thy-1.1 antigen and 2 membrane complement regulatory proteins (mCRPs), Crry and CD55, in three non-treated animals and the distribution of the injected antibody and C3 in the model was studied by immunohistochemistry. In the mesangial cells of the kidney, both expression of the antigen and distribution of the antibody with C3 deposition were observed with weak expression of mCRPs. There was also antigen and antibody distribution in the medullary cells of the adrenal gland and in the lymphocytes of the thymus but no C3 deposition, which was thought to be related to high expression of mCRPs. The antigen was observed in several other organs and tissues without distribution of the antibody. Cell death was only observed in the mesangial cells. These results clearly demonstrate that activation of CDC is regulated by several factors, such as distribution of the target molecule, antibody distribution and the balance among the molecules of the CDC cascade and mCRPs.     

Influence of the nitrogen form on in vitro organogenesis in Equisetum arvense

The physiological and biochemical mechanisms of organogenesis in Equisetum arvense have not been clarified yet. However, high concentrations of nitrogen have been shown to exert an inhibitory effect on in vitro tuber formation in E. arvense. The aim of this study was to clarify the influence of the form of nitrogen in a medium on in vitro organogenesis in E. arvense. Single‐node segments of E. arvense rhizomes were cultured in the test medium. The NH₄‐N and NO₃‐N concentrations of the test medium, respectively, were adjusted by adding NH₄H₂PO₄ and KNO₃ to the basal medium. The basal medium was a nitrogen‐free, modified form of White's medium. Vegetative shoots were newly formed in the test tubes for concentrations of NO₃‐N and NH₄‐N that exceeded 56 mg L^?1. However, no rhizome was formed at NH₄‐N concentrations exceeding 28 mg L^?1. The number of newly formed tubers decreased at an NH₄‐N concentration of 28 mg L^?1 and no tuber was formed at NH₄‐N concentrations exceeding 56 mg L^?1. In summary, although the presence of NO₃‐N in the medium did not inhibit in vitro rhizome or tuber formation in E. arvense, the presence of NH₄‐N in the medium exerted a strong inhibitory effect on the in vitro formation of both of these organs.     

Fusarium proliferatum, an additional bulb rot pathogen of Chinese chive

This is the first report of a disease of Chinese chive caused by Fusarium proliferatum. Because the symptoms are similar to those of the bulb rot (kampu-byo in Japanese) caused by F. oxysporum, we propose F. proliferatum as another causal agent of bulb rot of Chinese chive. Symptoms are wilting of leaves and brown rot on the basal bulbs of Chinese chive. A Fusarium sp., frequently isolated from symptomatic plants, produced identical symptoms on Chinese chive after inoculation, and was reisolated from the diseased plants. The fungus was identified as F. proliferatum based on morphological, cultural, and molecular characteristics.