Usefulness of the murine model to study the immune response against Histoplasma capsulatum infection

The present paper is an overview of the primary events that are associated with the histoplasmosis immune response in the murine model. Valuable data that have been recorded in the scientific literature have contributed to an improved understanding of the clinical course of this systemic mycosis, which is caused by the dimorphic fungus Histoplasma capsulatum. Data must be analyzed carefully, given that misinterpretation could be generated because most of the available information is based on experimental host–parasite interactions that used inappropriate proceedings, i.e., the non-natural route of infection with the parasitic and virulent fungal yeast-phase, which is not the usual infective phase of the etiological agent of this mycosis.     

Determination of the oxidative burst chemiluminescent response of avian and murine-derived macrophages versus corresponding cell lines in relation to stimulation with Salmonella serotypes.

In contrast to mammalian systems, avian species lack a resident or harvestable macrophage population in the abdominal exudate. Peritoneal macrophages in the chicken can be elicited if an inflammatory agent such as sephadex is injected. This study examines the kinetics of different macrophage populations, derived by different methods of isolation and from different hosts, with respect to the elicited oxidative burst upon infection with host-adapted Salmonella serotypes.

The nature of the oxidative burst elicited by murine and avian-derived and cell line macrophages was determined after stimulation with phorbol myristate (PMA), zymosan A, and Salmonella serotypes. Both murine and chicken peritoneal macrophages, chicken blood monocytes and corresponding cell lines, J774A.1 and HD-11, were unable to produce a detectable chemiluminescent (CL) response after interaction with Salmonella using the luminescent probe luminol. However, both PMA and zymosan A induced a CL response in all cell types, with PMA eliciting a higher and earlier peak response (pkH) than zymosan A. Lucigenin-enhanced CL in both murine and chicken macrophages was achieved with PMA, zymosan A and Salmonella serotypes. In this case, zymosan A induced higher responses than PMA. In the peritoneal macrophages of both hosts, there were no significant differences in the oxidative burst induced by the different Salmonella serotypes. However, the J774A.1 (murine) cells demonstrated significant differences, with S. enterica serotype Choleraesuis (S. choleraesuis and S. gallinarum producing the highest response. In the HD-11 (chicken) cells, S. choleraesuis and S. dublin elicited the higher CL. With both cell lines, S. abortusovis failed to induce an appreciable CL response.

In these experiments it was demonstrated that oxidative burst was not detectable in monocytes/macrophage populations using luminol, which suggests a link to the lack of a myeloperoxidase system in these cells. Lucigenin-enhanced CL appeared independent from the myeloperoxidase system, indicating production of another oxidative species compared with luminol. No discernable effect of host specificity with regard to Salmonella serotype and respective host was seen in host-derived or cell line macrophages, and cell line macrophages displayed altered functional characteristics with regard to oxidative burst in comparison with their primary counterparts.     

Research on Growth Behavior of Embryos for Bovine and Murine on Primary Murine Embryos Fibroblast Cell Feeder Layer

The difference in growth behavior between bovine embryos and murine embryos was studied on PMEF(primary murine embryos fibroblast)feeder layer. The results showed as follows: With embryos having attached, bovine embryonic trophoblast formed a transparent membranous structure covering on inner cell mass (ICM), however, murine embryonic trophoblast formed disc structure. Bovine embryos formed four kinds of ICM colonies with different morphology including the mass-like, the net-like, the stream-like and the mixture-like colonies. Compared with Murine ICM, the bovine ICM grew more fast. So, the bovine ICM was passaged at first after a culture of approximately 5 - 6 days in vitro, but murine ICM was passaged at first after an attachment of 3 - 4 days on PMEF feeder layer. The mixture colonies of bovine ICM differentiated very early, while the others differentiated very late. Most ICM-like mass of Bovine grew in a defined spot, but bovine ICMs like stream and ICMs like net proliferated fast and dispersed quickly. We found that the single blastomeres derived from late bovine morula and late murine morula formed sub-blastophere; moreover, the bovine ICM cell would differentiate rapidly if the trophoblast was removed.     

Research on Isolation and Clone of Embryonic Stem Cell-Like in Bovine

Bovine embryonic stem cell would be invaluable for researching the aspect of animal cloning, production transgenic animal and discussion of gene function in vitro. With the object of establishing an effective culture system for isolation and clone of bovine pluripotent stem cell, we cultured bovine embryos and mouse embryos including morula blastula and hatached blastula and obtained animal ICM on Primary marine embryonic fibroblast (Primary murine embryonic fibroblast, PMEF) feeder layer with tissue medium(DMEM supplemented with 15ml/100ml NBS ,0.1μmol/L Na2SeO3, 0. 1mmol/L β-mercaptoethanol, 1 000ng/ml LIF,10 ng/ml IGF, 1mmol/L necessary amino acid and 1mmol/L L-glutamine), then, we obtained mouse ICM and bovine ICM. Moreover, we isolated and cloned the 6 passage bovine ES like cells(12 cell lines) and 9 passage marine ES like cells (52 cell lines) deriving from bovine ICM and murine ICM respectively on the feeder layer of PMEF by disaggregating ICM and ES cell clones of bovine and murine into smaller clumps through digesting with 0. 125g/100ml trypsin and 0.02g/100ml EDTA and scattering with a glass needle. The pluripotency of both murine and bovine ES like cells was identified with morphological character, histochemistry identification, karyotype analysis and differentiation of ES cells in vitro or in vivo. This result showed that bovine embryonic stem cell and murine embryonic stem cell had developmental pluripotency.     

支持牛类胚胎干细胞发育的饲养层培养体系的建立

以小鼠胎儿和牛睾丸为材料 ,以含 1 5 % NBS、0 .1 mmol/ Lβ-巯基乙醇、0 .1 μmol/ L Na2 Se O3的DMEM溶液为培养液 ,分离获得了传 1 5代的小鼠胎儿成纤维细胞和 5代牛睾丸成纤维细胞 ,建立了小鼠和牛类 ES细胞培养体系。结果表明 :牛睾丸成纤维细胞和小鼠胎儿成纤维细胞均属附着生长型细胞 ,与小鼠胎儿成纤维细胞相比较 ,牛成纤维细胞直径和长度大 ,生长速度快 ,易于老化 ;1 2～ 1 6日龄的小鼠胎儿最适宜分离与克隆小鼠胎儿成纤维细胞 ;在 2 5℃条件下 ,以 0 .2 5 %胰蛋白酶 0 .0 4% EDTA消化液作用胎儿小块组织分离原代小鼠胎儿成纤维细胞 ,消化液作用时间不应超过 2 0 min,以相同的消化液在 3 7℃条件下 ,离散贴壁成纤维细胞 ,作用时间以 2～ 3 min为宜 ;培养细胞密度与传代时间间隔有密切关系 ,若传代时间间隔为 3～ 4d,培养细胞浓度应为 3× 1 0 5个 / ml～ 5× 1 0 5个 / ml;在成纤维细胞分离与克隆过程中 ,培养基中添加0 .1μmol/ L Na2 Se O3 0 .1 mmol/ Lβ-巯基乙醇 1 5 % NBS,有利于 MEF和 NBTF的增殖     

贺兰山西坡鼠种内脏重量与海拔梯度关系研究

采用铗日法对贺兰山西坡鼠类的内脏重量与海拔梯度关系情况进行了细致研究。选取8条典型沟道为样线,共布设有效铗日18 748个,捕获鼠类235只。阿拉善黄鼠和大林姬鼠在所有海拔梯度均有分布,其次是社鼠、灰仓鼠和短尾仓鼠,海拔分布范围最小的是高山鼠兔和东方田鼠。阿拉善黄鼠的肺脏和肾脏重量高海拔重量远高于低海拔重量,显示出了其对外界环境的超强适应能力。     

Cytotoxic activity of Origanum dictamnus

Several extracts of Origanum dictamnus, an endemic plant of Greece growing only in the island of Crete and the bioassay-directed isolated ursolic acid, were tested in vitro against the P388 (murine leukemia) and the human bronchial epidermoid cancer NSCLC-N6 (non small cell lung cancer) cell lines. Both the initial dichloromethane extract and the isolated from it ursolic acid exhibited cytotoxic activity. Ursolic acid was also tested in vivo, on murine ascite leukemia P388, where it exhibited at a dose of 50 mg/kg a marginal antileukemic activity.     

Cytotoxic resveratrol oligomers from the tree bark of Dipterocarpus hasseltii

A new resveratrol tetramer, named diptoindonesin E, was isolated from the acetone extract of the tree bark of Dipterocarpus hasseltii, together with five known resveratrol oligomers (-)-epsilon-viniferin, laevifonol, (-)-alpha-viniferin, vaticanol B, (-)-hopeaphenol, and a coumarin, scopoletin. The structures of these compounds were determined from spectroscopic evidence. Cytotoxicity test of the isolated compounds showed that hopeaphenol strongly inhibited murine leukemia P-388 cells.     

Determination of the oxidative burst chemiluminescent response of avian and murine-derived macrophages versus corresponding cell lines in relation to stimulation with Salmonella serotypes

In contrast to mammalian systems, avian species lack a resident or harvestable macrophage population in the abdominal exudate. Peritoneal macrophages in the chicken can be elicited if an inflammatory agent such as sephadex is injected. This study examines the kinetics of different macrophage populations, derived by different methods of isolation and from different hosts, with respect to the elicited oxidative burst upon infection with host-adapted Salmonella serotypes.The nature of the oxidative burst elicited by murine and avian-derived and cell line macrophages was determined after stimulation with phorbol myristate (PMA), zymosan A, and Salmonella serotypes. Both murine and chicken peritoneal macrophages, chicken blood monocytes and corresponding cell lines, J774A.1 and HD-11, were unable to produce a detectable chemiluminescent (CL) response after interaction with Salmonella using the luminescent probe luminol. However, both PMA and zymosan A induced a CL response in all cell types, with PMA eliciting a higher and earlier peak response (pkH) than zymosan A. Lucigenin-enhanced CL in both murine and chicken macrophages was achieved with PMA, zymosan A and Salmonella serotypes. In this case, zymosan A induced higher responses than PMA. In the peritoneal macrophages of both hosts, there were no significant differences in the oxidative burst induced by the different Salmonella serotypes. However, the J774A.1 (murine) cells demonstrated significant differences, with S. enterica serotype Choleraesuis (S. choleraesuis and S. gallinarum producing the highest response. In the HD-11 (chicken) cells, S. choleraesuis and S. dublin elicited the higher CL. With both cell lines, S. abortusovis failed to induce an appreciable CL response.In these experiments it was demonstrated that oxidative burst was not detectable in monocytes/macrophage populations using luminol, which suggests a link to the lack of a myeloperoxidase system in these cells. Lucigenin-enhanced CL appeared independent from the myeloperoxidase system, indicating production of another oxidative species compared with luminol. No discernable effect of host specificity with regard to Salmonella serotype and respective host was seen in host-derived or cell line macrophages, and cell line macrophages displayed altered functional characteristics with regard to oxidative burst in comparison with their primary counterparts.     

小鼠胎儿和牛睾丸成纤维细胞的冷冻保存

研究了冷冻保护液、平衡时间、降温速度、解冻液以及解冻方式对冷冻小鼠胎儿成纤维细胞和牛睾丸成纤维细胞活力的影响。结果表明,对小鼠胎儿成纤维细胞和犊牛睾丸成纤维细胞进行冷冻,冷冻液Ⅱ（０．４％ＢＳＡ）＋１．５ｍｏｌ／Ｌ乙二醇＋０．１ｍｏｌ／Ｌ蔗糖的ＰＢＳ液）的冷冻效果优于冷冻液Ⅰ（１０％的ＮＢＳ＋１０％ＤＭＳＯ的ＤＭＥＭ液）。用冷冻液Ⅱ对小鼠胎儿成纤维细胞和犊牛睾丸成纤维细胞进行冷冻,平衡时间可从２．