Changes in the distribution of integrins and their basement membrane ligands during development of human thyroid follicular epithelium

Interactions between cells and basement membrane components are crucial for the regulation of epithelial cell differentiation and polarization. We have studied by immunohistochemical methods the distribution of integrin adhesion proteins and some of their basement membrane ligands in foetal (16--19 weeks) and adult thyroid follicular epithelia. A diffuse immunoreactivity for only 3, v and 1 integrins was found in foetal follicular epithelium, whereas in adult follicular epithelium these integrins were expressed basally in a polarized manner. Additionally, 3 integrin was seen in a more basolaterally confined manner in adult follicular epithelium. Among basement membrane components, laminin 1, 1, 1 and 2 chains were found in epithelial basement membranes of the foetal thyroid gland, suggestive of the presence of laminins-1 and -3. In contrast, the basement membranes of adult follicular epithelium presented a much weaker immunoreactivity for the laminin 2 chain. Furthermore, immunoreactivity for the laminin 2 chain was occasionally seen in adult thyroid glands, apparently confined to myofibroblasts. Immunoreactivity for type IV collagen 1 and 2 (IV) chains was found in follicular basement membranes of foetal as well as adult thyroid gland. The results suggest that during maturation of foetal thyroid follicular epithelium a distinct polarization of integrins takes place. In mature thyroid follicular epithelium, the presumable adhesion-mediating integrin complexes are 31, v1 and/or v3 mediating adhesion to laminin-1 (1-1- 1) and type IV collagen trimer 12 (IV)     

Intracellular localization of factor VIII-related antigen and fibronectin in cultured human endothelial cells: evidence for divergent routes of intracellular translocation

Cultured human endothelial cells synthesize and secrete both fibronectin and factor VIII-related antigen (VIIIR:Ag). In immunofluorescence microscopy, intracellular fibronectin was seen diffusely perinuclearly whereas VIIIR:Ag was located both diffusely in the perinuclear cytoplasm and in distinct rod-shaped granules. These granules could, moreover, be visualized with fluorochrome-coupled Ricinus communis agglutinin I (RCA), which also stained the Golgi apparatus as a reticular juxtanuclear structure, and they were identified as Weibel-Palade bodies by immunoelectron microscopy. Puromycin treatment depleted intracellular fibronectin but did not affect the granular localization of VIIIR:Ag. A short exposure of the cells to monensin caused a juxtanuclear accumulation of fibronectin at the Golgi region whereas VIIIR:Ag only was seen in rounded cytoplasmic granules. A prolonged monensin treatment brought about a cytoplasmic accumulation of fibronectin-containing vesicles whereas VIIIR:Ag showed no accumulation and there was no codistribution between granules containing fibronectin or VIIIR:Ag. Type IV procollagen, on the other hand, was distinctly co-localized with fibronectin. In monensin-treated cells RCA mainly stained the VIIIR:Ag-containing vesicles whereas Concanavalin A (Con A) appeared to label the fibronectin-containing vesicles. Immunoelectron microscopy of these cells revealed VIIIR:Ag in some vacuolar structures and typical Weibel-Palade bodies could not be identified. Exposure of the cells to tunicamycin, on the other hand, caused a prominent cytoplasmic accumulation of VIIIR:Ag and, within 96 h, led to the disappearance of most of the VIIIR:Ag-positive granules but did not affect the intracellular distribution of fibronectin. These results, which show that metabolical inhibitors affect differently the intracellular compartmentalization of fibronectin and VIIIR:Ag, indicate, that the two glycoproteins have divergent intracellular pathways in cultured human endothelial cells.     

光刺激参数控制器

本文介绍一种能准确而迅速改变光刺激器光学参数(波长、强度等)的远距控制装置。将具有不同光学参数的滤光片(或孔径)置于转盘的不同位置,并对其编码。通过对编码值与预置值的比较,驱动步进马达带动转盘到所需的位置。仪器操作简便,有良好的重复性。     

Synthesis of monoacid 2,3-diacyl-sn-glycerols via 1,6-ditrityl-d-mannitol

Stereochemically pure 2,3-dipalmitoyl-sn-glycerol and 2,3-dioleoyl-sn-glycerol were prepared in an overall yield of 20% by a new and facile method starting from D-mannitol. The synthetic intermediates were 1,6-ditrityl-D-mannitol (1), 1-trityl-sn-glycerol (2), and 1-trityl-2,3-diacyl-sn-glycerol (3). The key reaction was the oxidation of 1 with lead tetraacetate followed by reduction with sodium borohydride. The product (2) was readily separated from the only byproduct, tritylethyleneglycol.     

Glial origin of rapidly adhering amniotic fluid cells.

Rapidly adhering cells (RA cells) from the amniotic fluid of a pregnancy with fetal anencephaly were investigated by immunofluorescence assay with an antiserum against glial cells. After 24 hours'' cultivation a high proportion of the cells showed positive glial-specific fluorescence, whereas no staining was seen in cells from samples of normal amniotic fluid. At the 24th week the mother was delivered of a stillborn infant with anencephaly. Immunofluorescence staining of RA cells with glial-specific antiserum may be used for the differential diagnosis of fetal abnormalities associated with a high alpha-fetoprotein concentration in amniotic fluid.     

A comparative analysis of intermediate filament proteins in bovine heart Purkinje fibres and gastric smooth muscle

The intermediate filament (IF) composition of muscle cells of various sources is still a controversial issue. In the present study, the IF composition of bovine Purkinje fibres (PFs), atrial and ventricular myocardium, and gastric smooth muscle (SM) has been compared using biochemical and immunocytochemical methods. The Mr of the major IF subunit protein in all four tissues was 55,000. In two-dimensional (2-D) electrophoresis gels of Triton-treated ordinary atrial and ventricular myocardium and the gastric muscular wall, two or three isoelectric isoforms were seen, whereas in PFs up to seven isoforms caused by phosphorylation were observed. In immunofluorescence studies antibodies against the Mr 55,000 subunit of PFs and gastric SM, respectively, both showed identical reactivity with PFs, atrial and ventricular myocytes, gastric SM cells and some SM cells in intramyocardial and gastric muscular wall blood vessels. A small amount of vimentin (Mr 57,000) was also detected in 2-D gel electrophoresis in all four tissues as well as in immunoblotting of PFs with antibodies to vimentin. Immunofluorescence studies using both polyclonal and monoclonal antibodies to vimentin showed that vimentin was present in the endothelium and SM cells of both intramyocardial and gastric muscular wall vessels, sometimes together with desmin in the vascular SM cells, but was never seen in PF, atrial, ventricular or gastric SM cells proper. As expected, vimentin was present in interstitial tissue, i.e., fibroblasts and capillaries. However, interestingly, the monoclonal antibodies, which recognized different antigenic determinants of vimentin, did not give identical staining patterns. Especially the staining of the vascular SM cells differed. Since this staining pattern did not change upon denaturation and unmasking experiments, it seems that the organization of vimentin in different mesenchymal cell types varies. Vimentin was also detected in isolated PFs but here it was located solely in the contaminating interstitial tissue. Thus, desmin is the sole IF protein expressed in PFs, in atrial and ventricular myocytes and in gastric SM cells proper; vimentin alone being present in the interstitial tissue cells, whilst in vascular SM cells desmin and vimentin are coexpressed in various proportions. The variation in number of isoforms of desmin and the heterogeneity in staining of mesenchymal tissues with monoclonal vimentin antibodies probably indicates that the IF cytoskeletons are differently organized in various cell types, even though they contain IFs of the same class.     

长裙竹荪抑菌活性物质的分离纯化及其抑菌效果

长裙竹荪Dictyophora indusiata是珍贵的食药用真菌,具有很强的抑菌作用,在天然防腐剂开发方面具有广阔的应用前景。本研究以长裙竹荪的抑菌活性为指标,通过萃取、3次不同流动相的硅胶柱层析、1次反相柱层析和薄层层析法对竹荪提取物进行分离纯化,得到一个抗菌活性强的单体化合物。根据核磁共振波谱等数据分析,推断该化合物为间苯三酚。以巨大芽孢杆菌和肠炎沙门氏菌为供试菌,用平板打孔法及原位抑菌法测定该化合物的抑菌效果,结果表明：该化合物对这两种菌有很强的抑制作用,半抑制浓度分别为83.06μg/mL和51.58μg/mL。本研究首次从长裙竹荪中获得具有抗菌活性的单体化合物间苯三酚,为竹荪天然抗菌物质的开发提供理论依据。