Immunocytochemical evaluation of epithelial–mesenchymal transition in epithelial tumors of dogs and cats

Epithelial–mesenchymal transition (EMT) plays a crucial role in metastasis of epithelial tumors; however, it is challenging to detect EMT by cytology. In the present study, EMT was visualized by fluorescence-immunocytochemistry (FICC). Air-dried smears from epithelial tumors of dogs (n=22) and cats (n=9) were stained using mouse monoclonal anti-E-cadherin and rabbit monoclonal anti‐vimentin antibodies. Enzymatic immunohistochemistry (IHC) revealed that 51.6% (8/22 in dogs, 8/9 in cats) of the cases showed EMT. In dogs, FICC could detect EMT in 62.5% (5/8) of those cases. In cats, FICC could detect EMT in 100% (8/8) of the cases. In conclusion, the present FICC method could successfully detect EMT using conventional air-dried cytology smear slides.     

Susceptibility to N-methyl-N-nitrosourea-induced retinal degeneration in different rat strains

To evaluate the potential role of genetic background in the susceptibility to retinal degeneration induced by N-methyl-N-nitrosourea (MNU), female rats of the Sprague-Dawley (SD), Long-Evans (LE) and Copenhagen (CH) strains were administered 50 mg/kg MNU or saline at 7 weeks of age. Retina morphology and morphometric analysis of all rats was performed 7 days after MNU administration. Atrophy of both the peripheral and central outer retina occurred in all rat strains exposed to MNU. Decreased photoreceptor cell ratio and increased retinal damage ratio were observed. The severities of the retinal atrophy were similar among all three rat strains. In conclusion, MNU-induced photoreceptor degeneration developed consistently in all three strains regardless of the absence (SD rats) or presence (LE and CH rats) of melanin in the retina, suggesting that genetic and melanin factors did not affect photoreceptor cell death after MNU.     

Factors Affecting the Development of Somatic Cell Nuclear Transfer Embryos in Cattle

Nuclear transfer is a complex multistep procedure that includes oocyte maturation, cell cycle synchronization of donor cells, enucleation, cell fusion, oocyte activation and embryo culture. Therefore, many factors are believed to contribute to the success of embryo development following nuclear transfer. Numerous attempts to improve cloning efficiency have been conducted since the birth of the first sheep by somatic cell nuclear transfer. However, the efficiency of somatic cell cloning has remained low, and applications have been limited. In this review, we discuss some of the factors that affect the developmental ability of somatic cell nuclear transfer embryos in cattle.     

Negative feedback regulation ensures the one receptor-one olfactory neuron rule in mouse

In the mouse olfactory system, each olfactory sensory neuron (OSN) expresses only one odorant receptor (OR) gene in a monoallelic and mutually exclusive manner. Such expression forms the genetic basis for OR-instructed axonal projection of OSNs to the olfactory bulb of the brain during development. Here, we identify an upstream cis-acting DNA region that activates the OR gene cluster in mouse and allows the expression of only one OR gene within the cluster. Deletion of the coding region of the expressed OR gene or a naturally occurring frame-shift mutation allows a second OR gene to be expressed. We propose that stochastic activation of only one OR gene within the cluster and negative feedback regulation by that OR gene product are necessary to ensure the one receptor-one neuron rule.     

Maintenance of mouse trophoblast stem cells in KSR-based medium allows conventional 3D culture

Mouse trophoblast stem cells (TSCs) can differentiate into trophoblast cells, which constitute the placenta. Under conventional culture conditions, in a medium supplemented with 20% fetal bovine serum (FBS), fibroblast growth factor 4 (FGF4), and heparin and in the presence of mouse embryonic fibroblast cells (MEFs) as feeder cells, TSCs maintain their undifferentiated, proliferative status. MEFs can be replaced by a 70% MEF-conditioned medium (MEF-CM) or by TGF-ß/activin A. To find out if KnockOut^TM Serum Replacement (KSR) can replace FBS for TSC maintenance, we cultured mouse TSCs in KSR-based, FBS-free medium and investigated their proliferation capacity, stemness, and differentiation potential. The results indicated that fibronectin, vitronectin, or laminin coating was necessary for adhesion of TSCs under KSR-based conditions but not for their survival or proliferation. While the presence of FGF4, heparin, and activin A was not sufficient to support the proliferation of TSCs, the addition of a pan-retinoic acid receptor inverse agonist and a ROCK-inhibitor yielded a proliferation rate comparable to that obtained under the conventional FBS-based conditions. TSCs cultured under the KSR-based conditions had a gene expression and DNA methylation profile characteristic of TSCs and exhibited a differentiation potential. Moreover, under KSR-based conditions, we could obtain a suspension culture of TSCs using extracellular matrix (ECM) coating-free dishes. Thus, we have established here, KSR-based culture conditions for the maintenance of TSCs, which should be useful for future studies.     

根区温度对水培生菜生长和矿质元素含量的影响

在水培条件下,研究了5个根区温度对生菜(Lactuca sativa L.)生长和矿质元素含量的影响。结果表明:随着根区温度的升高,生菜单株产量呈先增加后减少的趋势,在25℃时达到最大值,且干物质含量最高;根区温度对生菜叶片净光合速率和矿质元素含量的影响与对产量的影响相似,即在25℃时达到最大值。35℃根区温度处理下,根叶生长和地上部矿质元素的积累受到严重阻碍,生菜叶片硝酸盐含量显著下降。在5个根区温度处理中,25℃对水培生菜的生长及矿质元素的吸收最为适宜。     

Pigmentation of cultured red sea bream, Chrysophrys major, using astaxanthin from Antarctic krill, Euphausia superba, and a mysid, Neomysis sp.

Pigmentation enhancement in cultured red sea bream, Chrysophrys major, was investigated using Antarctic krill, Euphausia superba, and a mysid, Neomysis sp., as a source of astaxanthin. Diets fortified with processed Antarctic krill (krill meal) and its acetone extract, containing 0.82–4.92 mg carotenoids/100 g dry weight, and raw krill and raw mysid supplemented diets, containing about 2.00 mg carotenoids/100 g wet weight, were formulated and tested for carotenoid deposition. The rate of carotenoid deposition in fish fed with raw krill and raw mysid was significantly higher and resulted in distinct pigmentation. The groups fed with the krill meal and acetone extract diets showed varied concentrations of skin carotenoids and resulted in faint pigmentation. Pigmented fish then fed on a carotenoid-free diet for the same length of time showed no apparent differences in the skin pigmentation although the detectable amounts of carotenoids varied. The bream converted some of the dietary astaxanthin to skin tunaxanthin.