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.     

Anthracnose of sweet pepper caused by Colletotrichum scovillei in Japan

Severe fruit rot of sweet pepper was found in Shimane, Hyogo, Chiba, Toyama, and Nagano prefectures, Japan from 2005 to 2011. Dark, sunken spots with concentric rings of orange conidial masses appeared on fruits. Pathogenic isolates from diseased fruits in the prefectures were identified as Colletotrichum scovillei. This species was added to the pathogens of sweet pepper anthracnose in Japan. The representative isolate was pathogenic to sweet pepper, tomato and chili pepper fruits, kidney bean pod, azuki bean, pea and strawberry leaves, but a caused no symptoms on cucumber or carrot in inoculation tests.     

Nationwide seroprevalence of Neospora caninum among dairy cattle in Japan

Serum samples from 2420 clinically healthy dairy cattle, randomly selected from stored sera in 18 districts of Japan, were tested for the presence of Neospora caninum antibodies using an indirect fluorescent antibody test (titer > or =1:200). Nationwide seroprevalence is estimated at 5.7% (139/2420). Seropositive cattle were detected in all surveyed districts despite the evidence of confirmed case reports of bovine neosporosis, showing that N. caninum is widely distributed throughout Japan. Age-specific seroprevalence did not increase with cattle age, suggesting that Neospora infection is likely to be transmitted vertically rather than horizontally in Japan. Considering that N. caninum seropositive cows are thought to be more likely to abort, substantial fetal losses may be induced by N. caninum infection in Japan. Devising strategies are needed to reduce the economic impact on the Japanese dairy industry. This is the first study to investigate the nationwide seroprevalence of N. caninum in cattle in Asia.     

Canopy temperature on clear and cloudy days can be used to estimate varietal differences in stomatal conductance in rice

Leaf photosynthetic potential, a major determinant of yield potential in rice, is mostly explained by stomatal conductance (g_s) as well as leaf N content (Rubisco protein content). Therefore, the rapid and precise evaluation of g_s as well as leaf N content would be a valuable addition to rice breeding programs aimed at improved yield potential. We established a simple method based on infrared thermography to estimate varietal differences in g_s. Seven rice varieties were cultivated in two-row plots in 2007, and canopy temperatures in three varieties, including a control variety, Koshihikari, were simultaneously measured to determine canopy temperature difference (CTd) between Koshihikari and the other varieties on clear days. The varieties Takanari, Habataki, and TUAT1-5-6a displayed significantly (P < 0.05) lower leaf temperature and higher g_s and leaf photosynthetic rate (P_n) than Koshihikari. CTd was closely correlated with g_s and P_n. These results indicate that CTd may be useful as a relative index to estimate varietal differences in g_s. A significant CTd was still observed even under cloudy conditions in 2008. Takanari and TUAT1-5-6a had lower leaf temperature than Koshihikari on cloudy days as well as on clear days. Subsequent investigation of photosynthetic light response curves revealed that Takanari and TUAT1-5-6a had higher g_s and P_n than Koshihikari under lower irradiance. These results suggest that infrared thermography may be a simple method of evaluating varietal differences in g_s through CTd, and that it may be practicable even under cloudy conditions.     

Detection of chromogranin A in the adrenal gland extracts of different animal species by an enzyme-linked immunosorbent assay using Thomsen-Friedenreich antigen-specific Amaranthus caudatus lectin

The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen-Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.     

High levels of sugars and fructan in mature seed of sweet wheat lacking GBSSI and SSIIa enzymes

Sweet wheat (SW), which lacks functional granule-bound starch synthase I (GBSSI) and starch synthase IIa (SSIIa), accumulates high levels of free sugars in immature seeds. Here, we examined the effects of the lack of these two enzymes on mature kernel composition. Whole grain flour of SW had higher levels of sugars, particularly maltose, slightly higher ash and protein content, approximately two to three times higher lipid levels, and about twice as much total dietary fiber as parental or wild-type lines. Considerably higher levels of low-molecular-weight soluble dietary fiber (LMW-SDF), largely consisting of fructan, were also detected in SW. Although there were no differences in total amino acid levels, the free amino acid content of SW was approximately 4-fold higher than that of wild type, and the levels of certain free amino acids such as proline were particularly high. Thus, we were able to clearly demonstrate that the lack of GBSSI and SSIIa caused dramatic changes in mature seed composition in SW. These compositional changes suggest that SW flour may provide health benefits when used as a food ingredient.     

Specific and non-specific adsorption of inorganic ions I. Evaluation of specific adsorbability by means of minimum concentration for specific adsorption

The Minimum Concentration for Specific Adsorption (MCSA) was defined as a reversal of the charge concentration of the ion at the infinite dilution of sol (adsorbent), or practically at such concentration of sol that equilibrium of the ion Is but negligibly affected by adsorption of the ion.

The MCSAs of 15 species of cations were measured with respect to SiO₂ at pH 4,6,5, and 10; the MCSAs of 10 species of anions were measured with respect to Fe(OH)₃ at pH 4, and 6.5; and the MCSAs of 10 species of anions were measured with respect to Al-coated-SiO₂ at pH 6.5. Relative specificities of the inorganic ions based on the MCSAs at pH 6.5 were as follows:

The MCSAs of cations with respect to SiO₂ and of anions to Fe(OH)₃ and Al-coated-SiO₂ at pH 6.5 had close relationships to ionic potentials and general solubility of the elements in water. Based on these results, the authors concluded that the MCSA could be used u an index of specificity (i.e. covalent bond-forming tendency) of an Jon at adsorption reactions.

The values corresponding to the apparent stability constants of surface complexes formed by adsorption reaction with SiO₂ and cations at pH 4 were calculated from the values of the MCSAs.     

Metyltetraprole activity against plant pathogens with relatively rare cytochrome b haplotypes for azoxystrobin resistance

Journal of General Plant Pathology - Metyltetraprole is a novel quinone outside inhibitor (QoI) fungicide designed to avoid cross-resistance in cytochrome b G143A-harboring QoI-resistant...