Nonhomogeneous Deformation in the Anterior Leaflet of the Mitral Valve

In the mitral valve, regional variations in structure and material properties combine to affect the biomechanics of the entire valve. Previous biaxial testing has shown that mitral valve leaflet tissue is highly extensible, and exhibits nonlinear, anisotropic material properties. In this study, experimental measurements of mitral valve leaflet deformation under quasi-static pressure loading were performed on isolated porcine hearts. Biplane video images of markers placed on the anterior leaflet surface were used to reconstruct the 3D position of the markers at several pressure levels over the physiological range. A least-squares finite-element method was used to fit parametric models to the markers and to calculate the deformation over the surface. The results showed that the leaflet deformations were anisotropic, exhibiting a large nonhomogeneous radial stretch and a small circumferential stretch. This information can be used to better understand how the valve deforms under physiological loading, and to help design treatments for valve problems, such as mitral regurgitation.     

Activation of ion transport by combined effects of ionomycin,forskolin and phorbol ester on cultured HT-29cl.19A human colonocytes

The differentiated clone 19A of the HT-29 human colon carcinoma cell line was used as a model to study the intracellular electrophysiological effects of interaction of the cAMP, the protein kinase C (PKC) and the Ca²⁺ pathways, (a) A synergistic effect between ionomycin and forskolin was observed. From intracellular responses it was concluded that the synergistic effect is caused by activation of an apical Cl^– conductance by protein kinase A and a basolateral K⁺ conductance by Ca²⁺. (b) A transient synergistic effect of ionomycin and the phorbol ester phorbol dibutyrate (PDB) was found. The decrease of the response appeared to be due to PKC-dependent inactivation of the basolateral K⁺ conductance. The synergism is caused by PKC-dependent increase of the apical Cl^– conductance and Ca²⁺-dependent increase of the basolateral K⁺ conductance. (c) The effects of carbachol and PDB were not fully additive presumably because of their convergence on PKC activation, (d) Forskolin and PDB, when added in this order, had a less than additive effect. Results of cell-attached patch-clamp studies, presented in the accompanying paper, showed a synergistic effect of forskolin and PDB on non-rectifying small-conductance Cl^– channels. Assuming that these channels are involved in the transepithelial responses it is suggested that forskolin and PDB induce a modulatory, synergistic increase of the apical Cl^– conductance when both pathways are activated simultaneously. (e) The HT-29cl.19A cells differ from T₈₄ cells in that the latter did not respond with an increase of the short-circuit current to addition of phorbol ester. This may be due to a very low expression of PKC.     

Cholinergic responses in cloned human TE671/RD tumour cells

The cholinergic responses of the human tumour cell line TE671/RD were examined using digital Ca²⁺ imaging fluorescence microscopy and patch-clamp measurements. In response to stimulation of the muscarinic acetylcholine (ACh) receptor (mAChR), the intracellular concentration of Ca²⁺ ([Ca²⁺]_i) rose about two-fold, in parallel with inositol 1,4,5-trisphosphate accumulation, measured by chromatographic techniques. By contrast, there was no increment of [Ca²⁺]_i upon stimulation of the nicotinic ACh receptor (nAChR), nor after caffeine application. Electrophysiological experiments showed that TE671/RD cells lack functional voltage-activated Ca²⁺ channels. The stimulation of the nAChR induced transient whole-cell currents (I _ACh). Little or no current was detected in isotonic extracellular Ca²⁺, with Cs⁺ in the patch pipette. Cell pretreatment with muscarine reduced I _ACh by about 20%, without consistent modifications of current kinetics. Muscarine applied to the extra-patch membrane under the cell-attached configuration had no obvious effect on ACh-evoked unitary events. In conclusion, in human TE671/ RD cells, muscarinic stimulation increases [Ca²⁺]_i, while nicotinic stimulation does not. In addition, the nAChR exhibits peculiar ion permeability properties and is not functionally regulated by the breakdown of phosphoinositides.     

Establishment and characterization of the permanent human cell line C3842 derived from a secondary chondrosarcoma in Ollier’s disease

The permanent human cell line C3842 was established from a secondary chondrosarcoma in a typical case of Olliers disease. In the present study, we analyzed the morphological, cytogenetic and molecular biological characteristics of the cultured cells in comparison with the original tumor and investigated the invasion properties of the tumor model using functional imaging of proteolysis, matrigel assay and chick chorioallantoic membrane assay. C3842 cells exhibit the typical features of malignant cartilage tumor cells in vitro, including the expression of collagen types II, IX, XI and aggrecan. The proteolytic ability of C3842 cells is attributed to the expression of several proteases, such as cathepsin B, urokinase plasminogen activator and matrix-metalloproteinase-2, which enable the cells to degrade collagen type I and to permeate matrigel matrix. In accordance with the biological features in vivo, C3842 cells are not able to invade through the epithelium of the chick chorioallantoic membrane. In conclusion, the cell line C3842 provides the first model of a secondary chondrosarcoma in Olliers disease in vitro, which is characterized by distinct features of such malignant cartilage tumors.     

Establishment and characterization of cell line TNMY1 derived from human malignant fibrous histiocytoma

Although malignant fibrous histiocytoma (MFH) is one of the most common soft tissue sarcomas, its pathogenesis remains unclear. In this study, a cell line derived from human MFH, TNMY1, was established from a metastatic chest-wall lesion of a 60-year-old woman with MFH. The TNMY1 cell line was passaged 95 times, and it still retained the biological characteristics of the original tumor. TNMY1 consists of spindle-shaped cells and pleomorphic cells associated with multinucleated giant cells. Immunohistochemical studies showed that the spindle-shaped and pleomorphic cells were positive for vimentin, CD68 and alpha-smooth muscle actin, but negative for epithelial membrane antigen, desmin, muscle actin, alpha-sarcomeric actin, myoglobin, lysozyme and S-100 protein. The cells expressed collagen types I, III and V. These results indicate that MFH may originate from mesenchymal stem cells with the potential to differentiate into either fibroblasts or histiocytes. An elevated level of collagen type V mRNA expression is considered to support a diagnosis of MFH.     

人皮肤内神经-肥大细胞联接在经络线上的发现 Ⅱ“传入性”神经-肥大细胞联接和伴同传出性轴突的薛旺细胞

本研究取材于人的皮肤,电镜常规方法处理。发现人皮肤内有两种类型的神经肥大细胞联接。提示可能分别为传出性和传入性的,或暂称为A型和B型联接,分别报告于上篇和本篇中。 B型联接的构造特点是,脱开薛旺鞘样长带包被的轴突来到肥大细胞近旁,突进细胞体中。轴突终末的一部分被胞质唇封盖起来,终末周围的轴膜与细胞质膜形成联接的双层膜。无肥大细胞表面皱褶参与其形成,无线粒体和显明的囊泡存在终末之内,无薛旺细胞包被轴突终末。薛旺细胞伴同A型联接出现,可与肥大细胞紧密接触。胞质内有狭缝系统和粗面内质网片断,能拖出长尾,估计将过渡为鞘膜。薛旺细胞不是间织细胞,这里的间织细胞应是成纤维细胞。     

Alterations in Nonspecific Cross-reacting Antigen Localization during Cell Culture

Nonspecific cross reacting antigen (NCA), a constituent of the carcinoembryonic antigen family, was localized ultra-structurally in a human lung adenocarcinoma cell line, PC 9. NCA was distributed predominantly on the plasma membrane in the early phases of cell culture. Deletion of fetal bovine serum (FBS) from the culture medium suppressed cell division without significantly altering cell viability, and induced a dramatic but reversible change in NCA localization. Under these conditions, NCA was localized to membrane degradation products within cytoplasmic vesicles and vacuoles. Acid phosphatase activity was also present in some of these intracellular structures. Similar changes in NCA localization were seen in cells cultured with FBS at day 6 when the cells reached a plateau stage of growth. These findings strongly suggest that plasma membrane degradation is accelerated by the cessation of cell growth. Cytoplasmic reactivity for NCA in cancer cells may therefore reflect degradation of plasma membrane-associated NCA and may not necessarily be correlated with increased systhesis of this glycoprotein. Acta Pathol Jpn 39: 772 778, 1989.     

三磷酸腺苷对人胃癌细胞系MGC－803影响的电镜观察

用透射电镜和扫描电镜方法观察了三磷酸腺苷（ＡＴＰ）对人胃癌细胞系ＭＧＣ－８０３的影响，结果表明，ＡＴＰ能使ＭＧＣ－８０３细胞的核仁由网状向环状改变，微丝的组装得到改善，细胞表面微绒毛减少。提示ＭＧＣ－８０３细胞的恶性表型在ＡＴＰ作用下可向正常方向逆转。     

Immunoelectron microscopic localization of carcinoembryonic antigen in gastric adenocarcinoma cell lines

The distribution of carcinoembryonic antigen (CEA) in human gastric adenocarcinoma cell lines (HPE-GAC-3 cells and HPE-GAC-2 cells) was determined immunohistochemically by indirect peroxidase-labeled antibody method at the light and electron microscopic levels. In GAC-3 cells that proliferated as non-adherent single cells, CEA was located in the perinuclear spaces, the endoplasmic reticulum, Golgi apparatus, vesicles, multivesicular body (MVB) and entire plasma membrane. Membrane CEA was shown to be internalized into MVB in GAC-3 cells. In GAC-2 cells that form an acinus, CEA was predominantly present along the microvilli of the lumina) surface and in glycocalyceal bodies, the vesicles which bud from the microvilli into the lumen. These results suggest that in poorly differentiated cancer cells CEA is transported over the entire cell surface, retained on the membrane and accumulated Into the cell by way of the MVB, but in well differentiated cancer cells the newly synthesized CEA is rapidly and predominantly transported to the luminal surface and rapidly released from the membrane into the lumen by way of the glycocalyceal body.