A Cucumber mosaic virus Isolated from Silene armeria

A Cucumber mosaic virus was newly isolated from Silene armeria and was characterized by biological, serological and molecular biological methods. Received 4 July 2001/ Accepted in revised form 28 August 2001     

Colonic mast cell infiltration in rats with TNBS-induced visceral hypersensitivity

Colonic mucosal mast cells are implicated in the pathogenesis of visceral hypersensitivity associated with irritable bowel syndromes. This study was designed to investigate the roles of mucosal mast cells in development of an experimental visceral hypersensitivity induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in rats. TNBS, when injected into the proximal colon through laparotomy, produced a significant decrease in pain threshold of the distal colon to mechanical distention, indicating a visceral hypersensitivity. In the proximal colon that was directly insulted by TNBS, mucosal necrosis and extensive inflammatory cell infiltration were observed with concomitant increase in tissue myeloperoxide (MPO) activity. In the distal colon where distention stimuli were applied, the number of mucosal mast cells significantly increased following TNBS treatment, although neither mucosal injury nor increase in tissue MPO activity was observed. In an organ culture, spontaneous release of a mucosal mast cell-specific protease (RMCP-2) from the distal colon tissue of TNBS-treated rats was significantly larger than that of sham animals. Furthermore, TNBS-induced visceral hypersensitivity was significantly suppressed by subcutaneous pretreatment with a mast cell stabilizer doxantrazole in a dose-dependent manner. These findings suggest that prominent colonic mast cell infiltration associated with an enhanced spontaneous mediator release is responsible, at least partly, for development of visceral hypersensitivity induced by TNBS in rats.     

Effects of atelocollagen gel containing bone marrow-derived stromal cells on repair of osteochondral defect in a dog

To clarify the contribution of autologous transplantation of mesenchymal stromal cells (MSCs), an atelocollagen gel containing or not containing fluorescently-labeled canine MSCs was transplanted into an osteochondral defect which did not repair spontaneously and the histological repair of the defect was compared. Although an early repair of the cartilage was not observed in either defect, the reproduction of subchondral bone was remarkable in the MSCs-implanted defect. Moreover, in 2 weeks after operation, the implanted MSCs were located in the deeper regions of the defect, suggesting the differentiation of osteoblasts. There was a possibility that the movement of the implanted MSCs was due to an increase in intra-articular pressure from postoperative inflammation.     

Isolation and chemiluminescent properties of ferret (Mustela putorius furo) polymorphonuclear cells

Ferret polymorphonuclear cells (PMNs) and peripheral blood mononuclear cells (PBMCs) were separated from whole blood by density gradient centrifugation. Using a 50% Percoll solution (density=1.066), PMNs and PBMCs were successfully isolated after centrifugation; the purities of the PMNs and PBMCs were 94.2% and 95.6%, respectively. To evaluate the function of isolated ferret PMNs, we measured the superoxide generation with a MCLA-dependent chemiluminescence assay. The isolated ferret PMNs responded to phorbol 12-myristate 13-acetate (PMA) with kinetics similar to that of human PMNs. The ferret PMNs did not respond to N-formyl-Met-Leu-Phe (fMLF), unlike human PMNs, which rapidly responded. Thus, authors established a method for the rapid separation of highly purified populations of functional PMNs from the whole blood of ferrets.     

Phylogenetic characterization of rabies virus isolates from Carnivora in Brazil

The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.     

纤维素降解菌在高温厌氧消化过程中的作用

前期研究表明梭菌属的新株JC3（Clostridiumsp.Strain JC3）是嗜热产甲烷污泥中的主要纤维素降解菌。本项目采用分子生物学技术定量研究了菌株JC3在高温厌氧消化过程中的作用。在含三个小室（室1,室2,室3）的厌氧折流板反应器（9.5 L）中进行纤维素降解实验,试验温度设为55℃。当负荷为2.5 kg COD.m-3d-1,HRT为2天时,80%以上的COD进料都转化为甲烷。采用专门针对菌株JC3的特异性荧光针来研究折流式反应器中污泥样品。结果表明JC3细胞对DAPI染色细胞的比例从低于0.5%（最低检测极限）增加到9.4%（室1）,13.1%（室2）和21.6%（室3）。这表明由荧光原位杂交技术确定的菌株JC3细胞数与滞留污泥的纤维素降解产甲烷活性密切相关。设计了一个针对菌株JC3纤维素酶基因（纤维二糖水解酶A cbh A）的特异探针,并应用到处理固体废弃物如咖啡渣、废纸、垃圾和纤维素等消化污泥中,采用定量PCR技术发现菌株JC3细胞数与高温消化器中污泥的纤维素降解能力密切相关。因此,在高温厌氧消化过程中,菌株JC3对厌氧水解纤维素是非常重要的。