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.     

A new method for counting of quail leukocytes by flow cytometry

An automatic counting method was developed for fish blood cells using a fluorescent dye, 3, 3-dihexyloxacarbocyanine (DiOC (6)(3)), that selectively stain lipid bilayers in living cells. In the present study, the DiOC(6)(3) method was applied to quail (Cotumix cotumix japonica) blood cells. After quail blood cells were stained with DiOC(6)(3), absolute counts and relative proportions of erythrocytes, granulocytes, monocytes, and lymphocytes plus thrombocytes in whole blood were obtained by means of flow cytometry (FC). The number of each cell types by the FC was in good agreement with those counted microscopically. This method will offer new possibilities for routine blood cell counting for avian medicine.     

Annual changes in macrobenthos abundance in Funka Bay,Japan

The temporal variation in macrobenthos abundance was studied in Funka Bay in the periods 2001–2004 and 2007–2010. Dissolved oxygen just above the sea floor of the basin was lower and ignition loss at the level of the sea floor was higher in 2007–2010 than in 2001–2004. Macrobenthic species were categorized into three groups: long-lived species, sedental polychaeta Notomastus latericeus, and short-lived species. Ignition loss was the most influential factor affecting macrobenthos abundance as this variable restricts the recruitment of macrobenthos. The abundance of both long-lived species and N. latericeus was lower between 2007 and 2010 than between 2001 and 2004. Supplementation of oxygen from outside the bay by the Tsugaru Warm Current may be an important factor impacting the increase in the abundance of long-lived species. The abundance of N. latericeus barely increased between 2007 and 2010, possibly because dissolved organic matter, which is their food item, may have become limited due to a narrow chink between sediments with eutrophication. The abundances of short-lived species did not differ between the periods 2001–2004 and 2007–2010 due to the organisms having a low tolerance to hypoxia. The current conditions in Funka Bay are on the verge of turning the bay into an azoic zone.     

Growth,nutrient utilization,oxidative condition,and element composition of juvenile red sea bream <Emphasis Type="Italic">Pagrus major</Emphasis> fed with fermented soybean meal and scallop by-product blend as fishmeal replacement

A feeding trial was conducted to evaluate the effect of fermented soybean meal and scallop by-product blend (3:2) (FP) on the performance of juvenile red sea bream. Five isocaloric diets were prepared by replacing 0% (FP0), 15% (FP15), 30% (FP30), 45% (FP45), and 60% (FP60) fishmeal (FM) protein with FP, respectively. Triplicate groups of fish (initial mean weight 2.83 g) were fed the test diets for 45 days in a flowthrough seawater system. The results demonstrated that growth rates of fish fed FP0, FP15, and FP30 were similar, and significantly higher (P < 0.05) than those of FP45 and FP60. Nutrient utilization was significantly lower in FP60, with no differences found among the other groups. Dietary heavy-metal contents were affected by inclusion of FP, impacting on whole-body heavy-metal contents. In terms of oxidative stress, fish fed the FP30 diet were in the best condition, since this fish group showed the least oxidative-stressed condition as well as the highest tolerance against oxidation. In conclusion, the approach of utilizing this fermented mixture is promising and it could replace at least 30% FM protein in red sea bream diet without negative effects on performance, body composition or health of fish.     

Harpinpsta from Pseudomonas syringae pv. tabaci Is Defective and Deficient in Its Expression and HR-inducing Activity

To elucidate the role of harpins produced by Pseudomonas syringae, the corresponding hrpZ gene was isolated from P. s. pv. tabaci. The sequence information revealed that this gene carries a serious mutation with 326 bp lacking in the central region and potentially encodes only 140 N-terminal amino acids because of a frame shift. The investigation of biological properties using recombinant harpin indicated harpin_psta was incapable of inducing HR in both host and nonhost plants. Based on an immunoblot analysis to detect harpin from P. s. pathovars in hrp-inducing medium, the truncated harpin_psta was neither expressed nor secreted into the culture medium. These results suggest that harpin is not the sole determinant of the host-parasite specificity in P. s. pv. tabaci. Received 10 August 2000/ Accepted in revised form 21 December 2000     

Hindbrain decompression in a dog with scoliosis associated with syringomyelia

A 6-month-old female Border Collie was examined because of a 1-month history of progressive curvature of the cervical portion of the vertebral column. Radiography revealed severe cervical and thoracic scoliosis. Cervical syringomyelia and hydrocephalus were observed by means of magnetic resonance imaging. Suboccipital craniotomy and laminectomy of the first cervical vertebra were performed, and substantial improvement in the scoliosis and syringomyelia was observed 3 months after surgery. No recurrences were seen during the first year after surgery.     

Effect of setting conditions on mechanical properties of acid-induced Kamaboko gel from squid <Emphasis Type="Italic">Todarodes pacificus</Emphasis> mantle muscle meat

Squid Todarodes pacificus suwari gels, set at various temperatures and times, and acid-induced kamaboko gel, which was prepared by soaking suwari gel in 5% acetic acid for 20 h, were studied to evaluate the mechanical properties that are affected by setting conditions. Unset squid meat paste did not form a gel when soaked in acetic acid. The breaking strength of both suwari gel and acid-induced kamaboko gel showed a tendency to increase with setting temperature and time. SDS-PAGE analysis of suwari gel and acid-induced kamaboko gel, which were set at various temperatures and times, showed that myosin heavy chain (MHC) was observed at 30°C only for the first hour. The intensity of the MHC band at 30°C gradually decreased with setting time, while the intensity of the polymer band gradually increased with setting time. These results suggest that the protein-protein bonds in suwari gel affect the final texture of acid-induced kamaboko gel. Based on the analysis of the mechanical properties, and in consideration of the fact that the purpose of this experiment was to reduce energy usage, the best setting condition was determined to be 40°C for 3 h.