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Double Stained (double + stained)
Selected AbstractsEffects of hind limb denervation on the development of appendicular ossicles in the Dwarf African Clawed Frog, Hymenochirus boettgeri (Anura: Pipidae)ACTA ZOOLOGICA, Issue 4 2009Hyoung Tae Kim Abstract Sesamoids and other appendicular ossicles are common in other classes of vertebrates but comparatively rare in amphibians. The pipid frog Hymenochirus boettgeri (Boulenger, G. A. 1899. On Hymenochirus, a new type of aglossal batrachians. , Annals of the Magazine of Natural History Series 7: 122,125) is unusual among anurans in having seven (or more) appendicular ossicles in each hind limb. Sesamoids are often associated with muscles and tendons, and their development is usually regarded as mediated by or correlated with function. This study investigated the effects of paralysis (loss of function) on development of ossicles in the hind limb of Hymenochirus. Complete denervation of the right sciatic nerve was performed at developmental stages 63 and 66, and the animals maintained for a further 6,7 or 12,13 weeks. Specimens were cleared and double stained for cartilage and bone. There were no gross morphological differences between control and sham operated groups. The lunulae were not affected by paralysis, whereas the fabella arose later and/or regressed in some specimens. The distal os sesamoides tarsalia (OST) was shorter in paralysed individuals, and both the distal OST and cartilagines plantares showed delayed maturation. Denervation of the hind limb thus affected the timing of appearance, maintenance and rate of maturation of some sesamoid bones in Hymenochirus, but had no effect on others. [source] Expression of VCAM-1, ICAM-1, E-selectin, and P-selectin on endothelium in situ in patients with erythroderma, mycosis fungoides and atopic dermatitisJOURNAL OF CUTANEOUS PATHOLOGY, Issue 9 2000Vigfús Sigurdsson Background: Erythroderma may result from different causes. At present it is unclear whether the patho-mechanisms that lead to these different types of erythroderma are identical or different. Adhesion molecules and their ligands play a major role in endothelial-leukocyte interactions, which affect the binding, transmigration and infiltration of lymphocytes and mononuclear cells during inflammation, injury, or immunological stimulation. The aim of this study was to investigate the adhesion molecule expression on endothelial cells in erythroderma in situ. Methods: Snap-frozen skin biopsy specimens from 23 patients with erythroderma were studied. Eight had idiopathic erythroderma, 5 erythrodermic atopic dermatitis, 4 Sézary syndrome and 6 had erythroderma from miscellaneous causes. As a control we studied skin specimens from 10 patients with mycosis fungoides, 5 patients with atopic dermatitis and 5 healthy non-atopic volunteers. To determine adhesion molecule expression on endothelial cells in situ, sections were immuno-histochemically double stained with biotinylated Ulex Europaeus agglutinin 1 as a pan-endothelial cell marker, and for the adhesion molecules VCAM-1, ICAM-1, E-, and P-selectin. All double- and single-stained blood vessels in the dermis were counted. Results: Mean endothelial expression in erythroderma was as follows: VCAM-1 51.4%, ICAM-1 70.1%, E-selectin 43.5%, and P-selectin 52.6%. There was no statistical difference between different groups of erythroderma. Mean expression of all adhesion molecules tested, was in Sézary syndrome higher than in mycosis fungoides albeit not significant. In erythrodermic atopic dermatitis only VCAM-1 expression was significantly higher than in lesional skin of atopic dermatitis. No differences were observed in expression of the other three adhesion molecules. Conclusions: There is no difference regarding adhesion molecule expression on endothelial cells between different types of erythroderma. [source] Role of MMP9 on invadopodia formation in cells from adenoid cystic carcinoma.MICROSCOPY RESEARCH AND TECHNIQUE, Issue 2 2010Study by laser scanning confocal microscopy Abstract Migration, invasion and protease activity are essential for tumor progression and metastasis. Metastatic cells rely on invadopodia to degrade and invade extracellular matrix (ECM). Invadopodia are membrane protrusions with enzymes required for ECM degradation. These protrusions contain cortactin and membrane type 1 matrix metalloproteinase (MT1-MMP) superimposed to areas of digested matrix. Here we characterized invadopodia in a cell line (CAC2) derived from human adenoid cystic carcinoma. We carried out fluorescent-substrate degradation assay to assess in situ protease activity of CAC2 cells. Digestion spots in fluorescent substrate appear as black areas in green background. Cells were cultured on Matrigel-gelatin-FITC and fixed after 1 h and 3 h. CAC2 cells were double labeled to actin and cortactin. Cells were also double stained to actin and MT1-MMP. Samples were studied by laser scanning confocal microscopy. In all time points CAC2 cells showed actin, cortactin, and MT1-MMP colocalized with digestion spots in fluorescent substrate. We searched for other proteases involved in invadopodia activity. We have previously demonstrated that MMP9 influences adenoid cystic carcinoma behavior. This prompted us to investigate role played by MMP9 on invadopodia formation. CAC2 cells had MMP9 silenced by siRNA. After 1 h in fluorescent substrate, cells with silenced MMP9 showed clear decrease in matrix digestion compared with controls. No differences were found in cells with silenced MMP9 grown for 3 h on fluorescent substrate. Our results showed that CAC2 cells exhibit functional invadopodia containing cortactin and MT1-MMP. Furthermore, MMP9 would be required in the initial steps of invadopodia formation. Microsc. Res. Tech., 2010. © 2009 Wiley-Liss, Inc. [source] Interactive effects of cadmium and all- trans -retinoic acid on the induction of forelimb ectrodactyly in C57BL/6 mice,BIRTH DEFECTS RESEARCH, Issue 1 2006Grace S. Lee Abstract BACKGROUND Most toxicological studies have tested single chemical agents at relatively high doses, and fewer studies have addressed the toxic effects of chemical interactions. It is important to understand the toxicity of chemical mixtures in order to assess the more realistic risks of environmental and occupational exposures. A number of chemicals are known to induce a predominantly postaxial forelimb ectrodactyly in C57BL/6 mice, including acetazolamide, ethanol, cadmium, valproic acid, carbon dioxide, dimethadione, phenytoin, and 13- cis -retinoic acid and all- trans -retinoic acid (RA). In the present study, the interactive effects of coadministration of cadmium and RA on developing limbs were investigated. METHODS Pregnant C57BL/6 mice were treated with different intraperitoneal (IP) doses of cadmium chloride (CdCl2) and/or RA on gestational day (GD) 9.5, and fetuses were collected on GD 18 and double stained for examination of skeletal defects. RESULTS When RA was given simultaneously with cadmium, a significant increase in the incidence and severity of forelimb ectrodactyly (predominantly postaxial) was observed compared to the results with corresponding doses of cadmium or RA alone. When mice were exposed to subthreshold doses of both cadmium (0.5 mg/kg) and RA (1 mg/kg), the combined treatment exceeded the threshold, resulting in forelimb ectrodactyly in 19% of the fetuses. Moreover, coadministration of cadmium and RA at doses exceeding the respective thresholds showed a synergistic effect, that is, 92% of fetuses were found with the forelimb defect as opposed to 10% if the response were additive. CONCLUSIONS The findings demonstrate that concurrent exposure to these teratogens can have a synergistic effect and that subteratogenic doses may combine to exceed a threshold. Birth Defects Research (Part A), 2005. © 2005 Wiley-Liss, Inc. [source] Atlas of rat fetal skeleton double stained for bone and cartilageBIRTH DEFECTS RESEARCH, Issue 3 2001Elena Menegola Background The double staining of fetal skeleton for bone and cartilage is a very useful method to evidence skeletal abnormalities in laboratory animals. However, this method has been rarely used in routine developmental toxicity tests. One reason could be the difficulty of comparing the single skeletal pieces and of having reference points. In this paper the fetal rat skeleton double stained with Alizarin red S and Alcyan Blue is described in detail to produce an atlas for developmental toxicity laboratories. Teratology 64:125,133, 2001. © 2001 Wiley-Liss, Inc. [source] | |||||||||||||