MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

Strategies of newcastle disease vaccination for commercial ostrich farms in Japan

Strategies of Newcastle disease (ND) vaccination were demonstrated in a commercial ostrich farm in Japan. Three of 13 seven-month-old ostriches kept in a pen were vaccinated with a live ND vaccine by eye dropping for the 1st and 2nd vaccinations and spraying for the 3rd to 5th vaccinations. Antibodies against ND virus (NDV) were detected in all of the unvaccinated ostriches by virus neutralization test. At 2.5 months post final vaccination, 2 ostriches introduced into the pen raised antibodies against NDV. These data indicate that NDV may be transmitted from vaccinated to unvaccinated ostriches in the flock and that the virus may be sustained for a certain period in the flock. These data may be helpful for ND vaccination management in ostrich farms.     

Germination and appressorium formation of Pyricularia oryzae Cavara can be inhibited by reduced concentration of Blasin®Flowable with carbon dioxide microbubbles

We investigated the possibility to reduce the usage of Blasin^®Flowable (BF), a disinfectant inhibiting the germination and appressorium formation of Pyricularia oryzae Cavara conidia, by using carbon dioxide microbubbles (CO₂MB). Germination was significantly inhibited by 10 000-fold diluted BF solution containing CO₂MB generated by the decompression-type generator compared to CO₂ millibubbles (CO₂MMB) and CO₂MB generated by the gas-water circulating-type generator. Appressorium formation in the 10 000-fold diluted BF solution containing both CO₂MBs was less than that in CO₂MMB. Scanning electron microscopy showed wrinkles and dents on the surface of conidia treated with 5 000-fold diluted BF solution containing both CO₂MBs. Via transmission electron microscopy, we observed the expansion of the vacuole and the intracellular space and bloated or absent lipid granules in the conidia treated with BF solution containing both CO₂MBs. Our results show that inhibition of the conidium germination and appressorium formation of P. oryzae Cavara by 10 000-fold diluted BF solution could be achieved by using the decompression-type CO₂MB.     

Oxygen isotopic compositions of asteroidal materials returned from Itokawa by the Hayabusa mission

Meteorite studies suggest that each solar system object has a unique oxygen isotopic composition. Chondrites, the most primitive of meteorites, have been believed to be derived from asteroids, but oxygen isotopic compositions of asteroids themselves have not been established. We measured, using secondary ion mass spectrometry, oxygen isotopic compositions of rock particles from asteroid 25143 Itokawa returned by the Hayabusa spacecraft. Compositions of the particles are depleted in (16)O relative to terrestrial materials and indicate that Itokawa, an S-type asteroid, is one of the sources of the LL or L group of equilibrated ordinary chondrites. This is a direct oxygen-isotope link between chondrites and their parent asteroid.     

Anthracnose of Madagascar periwinkle caused by species belonging to the Colletotrichum gloeosporioides species complex

Severe blight of stems was found on Madagascar periwinkle (Catharanthus roseus) in Shizuoka and Okinawa prefectures, Japan, in July 2003 and September 2004, respectively. Similar fungi were isolated from the diseased plants. The isolated fungi caused the disease after inoculation and subsequently were reisolated from diseased tissues. The fungi were identified as species closely related to C. siamense or C. tropicale according to sequences of the β-tubulin-2 gene, indicating that they belong to the Colletotrichum gloeosporioides species complex. This is the first report specifying the genetic relationships of the pathogens of Madagascar periwinkle anthracnose in the complex.     

Identification of a bacterium isolated from galls on carrot and weeds

In 2004, bacterial galls were found on the roots of carrots in Shizuoka Prefecture, Japan. Galls were about 0.1–2 cm in diameter, light brown in color and had rough surfaces. In 2005, similar galls were found on the roots of three weeds: henbit (Lamium amplexicaule L.), Persian speedwell (Veronica persica Poir.) and leaf mustard (Brassica juncea L.). A bacterium that forms white, rough colonies was isolated from the carrot and weeds galls. The bacterial isolates had properties identical with Rhizobacter dauci Goto and Kuwata. Phylogenetic analysis based on 16S rDNA sequences showed that the carrot isolate had the highest homology (similarity of 100%) with that of the type strain of R. dauci. Rep-PCR genomic fingerprinting using BOX A1R primer showed that the carrot and weeds isolates were nearly identical. Pathogenicity of the isolates was confirmed by inoculating the roots of carrots and the weeds. After 2–5 weeks, they formed galls on the roots of the original host species and on other plant species tested. The galls were indistinguishable from those formed naturally, and the inoculated bacterium was reisolated. Thus, the causal bacterium of carrot and weeds gall was identified as R. dauci, and the bacterium was found to have a wider host range than previously known. These weed hosts may serve as inoculum sources for carrot bacterial gall disease.     

Marigold Leaf Spot Caused by Alternaria tagetica New to Japan

In October 1998, a disease causing mainly foliar necrotic lesions was found on African marigold (Tagetes erecta) and French marigold (T. patula) grown in Miyagi Prefecture, Japan. Similar lesions also developed on stems and flowers, resulting in early blight of the affected organs. Plants with numerous lesions withered rapidly. A mitosporic fungus isolated repeatedly from the diseased plants was identified as Alternaria tagetica and demonstrated to cause the disease. The disease, as well as the fungus, is new to Japan. We propose the name “hanten-by?”, which means leaf spot in Japanese, for this disease. Received 5 April 2000/ Accepted in revised form 31 May 2000