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 bronchopulmonary artery fistula in a horse

A 24-year-old, Thoroughbred gelding presented with difficulty breathing for a few days and intermittent nose bleeding before dying. At necropsy, the bronchoesophageal artery and the bronchial artery that flowed into the left anterior lobe were tortuous and dilated, and it was found that dilated tortuous branches of the bronchial artery ran over the dorsal and ventral surfaces of the left anterior lobe. Histopathologically, an anastomosis between a muscular artery and an elastic artery were demonstrated, which were identified as bronchial and pulmonary arteries, respectively. Based on the gross and histopathological findings, a bronchopulmonary artery fistula was diagnosed. To the best of our knowledge, this is the first case report of a pulmonary vascular anastomosis in a horse.     

Association of BoLA-DRB3 alleles identified by a sequence-based typing method with mastitis pathogens in Japanese Holstein cows

The association of the polymorphism of bovine leukocyte antigen ( BoLA-DRB3 ) genes identified by the polymerase chain reaction sequence-based typing (PCR-SBT) method with resistance and susceptibility to mastitis caused by pathogenic bacteria was investigated. Blood samples for DNA extraction were collected from 194 Holstein cows (41 healthy cows and 153 mastitis cows including 24 mixed-infection cows infected with 2 or 3 species of pathogens) from 5 districts of Chiba prefecture, Japan. Sixteen BoLA-DRB3 alleles were detected. The 4 main alleles of DRB3*0101 , *1501 , *1201 , and *1101 constituted 56.8% of the total number of alleles detected. Mastitis cows were divided into 2 groups: group 1 with single-infection cows and group 2 with all mastitis cows including 24 mixed-infection cows. The differences in the frequencies of BoLA-DRB3 alleles and the number of cows homozygous or heterozygous for each BoLA-DRB3 allele between healthy cows and the 2 groups of mastitis cows were evaluated. Furthermore, similar comparisons were performed between healthy cows and the 2 groups of mastitis cows for each mastitis pathogen. It was considered that the 4 alleles, namely, DRB3*0101 , *1501 , *1201 , and *1101 had specific resistance and susceptibility to 4 different mastitis pathogens. Thus, DRB3*0101 might be associated with susceptibility to coagulase-negative Staphylococci and Escherichia coli , and DRB3*1501 might be associated with susceptibility to Escherichia coli . However, DRB3*1101 might be associated with resistance to Streptococci and coagulase-negative Staphylococci , and DRB3*1201 , with resistance to Streptococci , Escherichia coli , and Staphylococcus aureus .     

Do Horses Prefer Certain Substrates for Rolling in Grazing Pasture?

We assessed whether rolling damage by grazing horses could be reduced by constructing areas assigned for rolling. A group of horses were enclosed in a paddock with and without rolling areas made of dry soil, sand, and straw. Their behavior was recorded for 1 week in the paddock without any treatment (control paddock). Then the horses were moved to another paddock with the rolling areas (rolling paddock). After a 3-week familiarization period, horses were observed for 1 week. In the rolling paddock, the frequency and time spent rolling were significantly greater in rolling areas than in nonrolling areas. Horses significantly preferred the soil rolling area than sand and straw (P < .05). Although rolling was considered the most relevant body care behavioral element, the effects of the substrate in rolling areas on other body care behavioral activities, such as mutual and self-grooming, also were investigated. The frequency and duration of mutual grooming and the duration of self-grooming decreased significantly in the rolling paddock compared with the control paddock (P < .05). Hence, offering a rolling area encourages horses to roll in these areas and keeps the pasture in good condition; therefore, grazing time can be increased, with less reliance on supplementary feed.     

Pathological and phylogenetic features of prevalent canine distemper viruses in wild masked palm civets in Japan

Ten wild masked palm civets infected with canine distemper virus (CDV), captured in Japan from 2005 to 2007, were histopathologically and phylogenetically analyzed. Phylogenetic analysis based on the amino acid sequences of the H protein of two CDV isolates from masked palm civets revealed that the two isolates were classified into the clade of recent isolates in Japan. Histopathologically marked lesions of virus encephalitis were present in the brain, whereas gastrointestinal lesions were absent or at a mild degree. The distribution of the lesions resembles that of recent CDV cases in dogs. Therefore, recent CDV infections in masked palm civets could be caused by recently prevalent CDV in dogs. The possibility of the masked palm civet as a spreader of CDV among wildlife is also discussed.     

Immunostimulatory effects of natural human interferon-alpha (huIFN-α) on carps Cyprinus carpio L.

Human interferon-α (huIFN-α) is an important immunomodulatory substance used in the treatment and prevention of numerous infectious and immune-related diseases in animals. However, the immunostimulatory effects of huIFN-α in fish remain to be investigated. In the current study, the immune responses of the carp species Cyprinus carpio L. to treatment with huIFN-α were analyzed via measurement of superoxide anion production, phagocytic activity and the expression of cytokine genes including interleukin-1β, tumor necrosis factor-α and interleukin 10. Low doses of huIFN-α were administered orally once a day for 3 days, and sampling was carried out at 1, 3 and 5 days post-treatment. Our results indicate that a low dose of huIFN-α significantly increased phagocytic activity and superoxide anion production in the carp kidney. The huIFN-α-treated fish also displayed a significant upregulation in cytokine gene expression. The current study demonstrates the stimulatory effects of huIFN-α on the carp immune system and highlights the immunomodulatory role of huIFN-α in fish.     

Improved porcine model for Shiga toxin‐producing Escherichia coli infection by deprivation of colostrum feeding in newborn piglets

Porcine edema disease (ED) is a toxemia caused by enteric infection with Shiga toxin 2e (Stx2e)‐producing Escherichia coli (STEC). ED occurs most frequently during the weaning period and is manifested as emaciation associated with high mortality. In our experimental infection with a specific STEC strain, we failed to cause the suppression of weight gain in piglets, which is a typical symptom of ED, in two consecutive experiments. Therefore, we examined the effects of deprivation of colostrum on the sensitivity of newborn piglets to STEC infection. Neonatal pigs were categorized into two groups: one fed artificial milk instead of colostrum in the first 24 h after birth and then returned to the care of their mother, the other breastfed by a surrogate mother until weaning. The oral challenge with 10¹¹ colony‐forming units of virulent STEC strain on days 25, 26 and 27 caused suppression of weight gain and other ED symptoms in both groups, suggesting that colostrum deprivation from piglets was effective in enhancing susceptibility to STEC. Two successive STEC infection experiments using colostrum‐deprived piglets reproduced this result, leading us to conclude that this improved ED piglet model is more sensitive to STEC infection than the previously established models.