Effect of intramammary injection of rboGM-CSF on milk levels of chemiluminescence activity, somatic cell count, and Staphylococcus aureus count in Holstein cows with S. aureus subclinical mastitis

The effect of intramammary injection of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF, 400 microg/10 mL) on quarter milk levels of chemiluminescence (CL) activity, and somatic cell count (SCC) and shedding pattern of Staphylococcus aureus was investigated. Ten Holstein cows, naturally infected with S. aureus were used, with either early-stage or late-stage subclinical mastitis. Injection of rboGM-CSF caused a remarkable increase in milk CL activity with a peak at 6 h after the cytokine injection in the early- and late-stage groups. In the early-stage group, milk SCC stayed around preinjection level at 6 h, rose significantly on days 1 and 2, and was followed by a smooth and significant decline to an under preinjection level (below 200 000 cells/mL) on day 7 postinjection. Alternatively, in the late-stage group, milk SCC rose significantly at 6 h after the cytokine injection and maintained high levels thereafter. The milk S. aureus count decreased drastically by the cytokine injection in the early-stage group. The bacterial count was moderately decreased in the late-stage group, but increased back to preinoculation levels on day 7 after the cytokine injection. The results suggest that the rboGM-CSF has a potential as a therapeutic agent for S. aureus infection causing subclinical mastitis of dairy cows, if the cytokine is applied at the initial stage of infection.     

我国三种野生泥鳅遗传多样性的微卫星分析

本研究采用5个微卫星分子标记技术对我国黑龙江泥鳅(M.mohoity)、北方泥鳅(M.bipartitus)、泥鳅(M.anguillicaudatus)进行了遗传多样性分析。结果表明,三种泥鳅的5个微卫星基因座共检测到等位基因数27个,微卫星基因座的等位基因数范围为3~7个。5个微卫星基因座在三种泥鳅的多态信息含量都达到了高度多态的水平,平均PIC值在0.526~0.722之间,说明三种泥鳅具有丰富的遗传多样性。黑龙江泥鳅、北方泥鳅和泥鳅的观测杂合度Ho分别为0.978,0.889和0.742,期望杂合度He分别为0.683、0.781、0.721。三种泥鳅的遗传距离为0.189~0.559,泥鳅和北方泥鳅群体间遗传距离最小为0.189,亲缘关系较近,而泥鳅和黑龙江泥鳅群体间遗传距离最大为0.559,亲缘关系较远。Hardy-Weinberg平衡检验P值。得出所有个体在5个微卫星位点上都无偏离现象。聚类结果显示黑龙江泥鳅、北方泥鳅、泥鳅群体分为二支,泥鳅和北方泥鳅聚为一类,之后两者再与黑龙江泥鳅聚为一类。     

Comparison of dendritic cell-mediated immune responses among canine malignant cells

Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials.     

The influence of barley malt protein modification on beer foam stability and their relationship to the barley dimeric alpha-amylase inhibitor-I (BDAI-I) as a possible foam-promoting protein

The foam stability of beer is one of the important key factors in evaluating the quality of beer. The purpose of this study was to investigate the relationship between the level of malt modification (degradation of protein, starch, and so on) and the beer foam stability. This was achieved by examining foam-promoting proteins using two-dimensional gel electrophoresis (2DE). We found that the foam stability of beer samples brewed from the barley malts of cultivars B and C decreased as the level of malt modification increased; however, the foam stability of cultivar A did not change. To identify the property providing the increased foam stability of cultivar A, we analyzed beer proteins using 2DE. We analyzed three fractions that could contain beer foam-promoting proteins, namely, beer whole proteins, salt-precipitated proteins, and the proteins concentrated from beer foam. As a result, we found that in cultivar A, some protein spots did not change in any of these three protein fractions even when the level of malt modification increased, although the corresponding protein spots in cultivars B and C decreased. We analyzed these protein spots by peptide mass finger printing using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. As a result, all of these spots were identified as barley dimeric alpha-amylase inhibitor-I (BDAI-I). These results suggest that BDAI-I is an important contributor to beer foam stability.     

Spontaneous well-differentiated pancreatic islet cell carcinoma with vascular invasion in a male F344 rat

This case report describes a case of spontaneous pancreatic islet cell carcinoma with vascular invasion in a 110-week-old male F344 rat. Histologically, a pancreatic nodule consisting of tumor cells and many blood-rich vessels, and covered with a fibrous capsule showed local invasion in the capsule and adjacent acinar tissues, encircling a large duct-like structure (DS). The tumor was composed of well-differentiated tumor cells resembling normal pancreatic islet cells, which had small round nuclei and eosinophilic cytoplasm. Mitotic figures were rare. Immunohistochemistry revealed that the tumor cells were positive for insulin. Although endothelial cells were not detected, the DS wall showed cells positive for α-smooth muscle actin and elastic fibers, suggesting that the DS is the pancreatic artery. This is a rare case of islet cell carcinoma consisting of well-differentiated tumor cells with invasion of the pancreatic artery in a rat.     

Malignant pinealoma observed in the deep cerebral parenchyma of a male Wistar rat

This report describes a case of spontaneous malignant pinealoma in a 90-week-old male Wistar rat. The tumor mass occurred in the deep cerebral parenchyma and no intact pineal gland was observed in the area between the posterior-dorsal median line of the cerebrum and the cerebellum. The tumor was characterized by a large nodular proliferation occupying the central area of the brain, extending from the dorsal surface to the base of the brain, corresponding to the thalamus. The tumor cells had round to irregular oblong nuclei approximately 5–17 μm in diameter and showed faintly or moderately eosinophilic cytoplasm and indistinct cell boundaries. Immunohistochemically, the tumor cells were positive for synaptophysin and partially positive for neuron-specific enolase (NSE). The tumor showed malignant features including cellular pleomorphism, high mitotic index, necrotic foci, and invasive and extensive growth and was, therefore, diagnosed as an extremely rare malignant pinealoma in the deep cerebral parenchyma.     

A possible critical dosing period of p-cumylphenol for development of cystic kidneys in rat neonates

In accordance with a previous report on cystic kidneys induced in rat neonates when dosed with p-cumylphenol (PCP) for 18 days from postnatal day (PND) 4, 3 rat neonates were dosed with PCP once a day for 14 days, either from PND 14, 21, 28, 35, or 42 as W2, W3, W4, W5, and W6 groups, respectively, to investigate whether dosing periods in different PNDs influenced the development of cystic renal tubules. The lesion was striking in the W2 group and at a lesser magnitude in the W3 group, whereas either kidney was unaffected when dosing was initiated beyond PND 28. These findings, together with the results from the previous study, suggested that PND 14-28 is a critical dosing period for PCP to develop cystic kidneys in rat neonates. The lining epithelium of the cystic tubules was immunohistochemically positive for AQP2. This finding and the anatomical location indicated that the cystic tubules were of collecting duct origin. Either obstruction, fluid accumulation, or reparative hyperplasia of the lining epithelium was unlikely to be involved in the formation of cystic tubules lined with a monolayer of cuboidal or columnar epithelium with a high nuclear density. Thus, the follow-up investigation on PCP suggested a critical dosing period of PND 14-28 in rat neonates for the development of cystic dilation of renal collecting ducts. This study further supports that additive hyperplasia of the lining epithelium is a fundamental basis of this unique lesion.