High-voltage-activated calcium channel messenger RNA expression in the 140-3 neuroblastoma-glioma cell line

Expression of calcium channel alpha1 subunits in oocytes or cell lines has proven to be a powerful method in the analysis of structure-function relations, but these experimental systems are of limited value in the examination of neuron-specific functions such as transmitter release. Cell lines derived from neurons are often capable of these functions, but their intrinsic calcium channel alpha1 subunits are complicating factors in experimental design. We have examined the biophysical and molecular properties of calcium channels in a little studied neuroblastoma-glioma hybrid cell line, 140-3, a close relative of the NG108-15 cell line, to test whether this cell line might serve a role as an expression system for neural mechanisms. This cell was selected as it contains an intact transmitter release mechanism yet secretes little in response to depolarization. Patch-clamp recording revealed only a prominent low-threshold, rapidly inactivating calcium current with a single-channel conductance of approximately 7 pS that was identified as T type. A search for calcium channel alpha1 subunit messenger RNA in the 140-3 cells with three different tests only revealed alpha1C, whereas alpha1A-alpha1C were present in the parent NG108-15 line. We made a particular effort to search for alpha1E, since this subunit has been associated with a low-voltage-activated current. Our findings suggest that, since the principal nerve terminal-associated calcium channels (alpha1A, alpha1B, alpha1E) are absent in the 140-3 cell, this cell line may prove a particularly useful model for the analysis of the role of high-voltage-activated calcium channels in complex functions of neuronal cells.     

Technical evolution of arthoscopic knee surgery

The widespread growth of arthroscopic techniques and their use has dramatically changed the practice of orthopaedic surgery. A high degree of clinical accuracy and minimally invasive procedure with a low surgical morbidity have encouraged the use of arthroscopy to assist in diagnosis, to determine prognosis and to provide treatment. In particular, the knee is the proper joint in which arthroscopy has its diagnostic and intraarticular surgical application. The rapid advancement of arthroscopic techniques has demonstrated a variety of surgical techniques in procedures such as meniscectomy, meniscal repair and cruciate ligament reconstruction during the last decade. This article reviews the past history of arthroscopy and technical evolution of arthroscopy in knee surgery in our department.     

Endoscopic surgery for obstructive hydrocephalus

Endoscopic surgery is popular in the neurosurgical field. The purpose of this study was to determine the role of endoscopy in obstructive hydrocephalus. From 1989 to 1999, we performed 81 endoscopic third ventriculostomies and 10 septostomies. Seventy-one of 81 operations were performed with endoscopic third ventriculostomy alone and 10 patients had endoscopic third ventriculostomy and ventriculoperitoneal shunt simultaneously. Age distribution varied from 2 months to 62 years of age. Our selection criteria included aqueductal stenosis (39 patients) and obstructive hydrocephalus due to tumor or cyst (42 patients). The most common candidate for endoscopic septostomy was atresia of the foramen of Monro (4 patients). Endoscopic septostomy was also performed to simplify shunting in patient; with multiseptated ventricle due to shunt infection, germinoma, thalamic tumor, craniopharyngioma, cyst and brain abscess. Sixty-five of 71 patients who were treated with endoscopic third ventriculostomy alone showed successful results (91.5%). However, 6 patients had unsatisfactory results and they needed a ventriculoperitoneal shunt. With no mortality, transient surgical complications were observed in 7 patients: 2 transient diabetes insipidus from electrical injury to the pituitary stalk, 1 epidural hematoma from sudden drainage of CSF, 1 delayed intraventricular hemorrhage. 2 obstruction of fenestration site and 1 transient memory disturbance from injury to the fornix. Endoscopic septostomy was useful in simplifying shunting in all cases with complicated hydrocephalus. Endoscopic surgery is straightforward and effective in appropriately selected cases with obstructive by drocephalus.     

Immunohistochemical analysis of Fas ligand expression in normal human tissues

Cross-linking of Fas and Fas ligand (FasL) induces apoptosis in Fas-bearing cells and regulates apoptosis. Fas is widely expressed in normal human tissues, but FasL expression has been considered to be restricted to lymphoid tissues. Recent studies have demonstrated that FasL is also expressed in some nonlymphoid tissues. To screen the in situ expression of FasL in normal human tissues, immunohistochemistry was performed using paraffin-embedded human tissues. FasL immunostaining was easily detected in testis, neurons, trophoblasts, tonsil, lymph node, Paneth cells, hepatocytes, renal tubular epithelium and bronchial epithelium, consistent with previous reports. Surprisingly, FasL was also expressed in many other cell types, including thymic medulla, skeletal muscle, cardiac muscle, pituitary gland, parathyroid gland, prostate glands, oocytes, epithelium of fallopian tube, endometrial glands, and gastric parietal cells. These findings demonstrate that FasL is widely expressed in human tissues and suggest that wide but cell-type specific expression of FasL may not only be implicated in the regulation of immune homeostasis but also in the regulation of cell death and life in many cell types in vivo.     

Immunolocalization of CD1d in human intestinal epithelial cells and identification of a beta2-microglobulin-associated form

In order to better understand the role of intestinal CD1d, we sought to define the cellular localization and further characterize the biochemical structure of CD1d in human intestinal epithelial cells (IEC). Using a CD1d-specific rabbit anti-gst-CD1d antibody, immunoprecipitation of radiolabeled cell surface proteins detected a previously identified 37 kDa protein as well as a 48-50 kDa protein which were confirmed by Western blotting with a CD1d-specific mAb, D5. Immunoprecipitation of protein lysates with the CD1d-specific mAb, D5 and 51.1.3, and the beta2-microglobulin (beta2m)-specific mAb, BBM.1, followed by N-glycanase digestion and Western blotting with the D5 mAb showed that the 48-50 kDa protein was a beta2m-associated, CD1d glycoprotein. CD1d was immunolocalized to the apical and lateral regions of native small and large intestinal IEC as defined by confocal laser microscopy using the D5 mAb and the rabbit anti-gst-CD1d antibody. In addition, a large apical intracellular pool of CD1d was identified. Identical observations were made with polarized T84 cells. Selective biotin labeling of apical and basolateral cell surfaces followed by immunoprecipitation with the D5 mAb, N-glycanase digestion and avidin blotting confirmed the presence of glycosylated CD1d on both cell surfaces and immunolocalization of the 37 kDa non-glycosylated form of CD1d to the apical cell surface. These studies show that CD1d is located in an ideal position for luminal antigen sampling and presentation to subjacent intraepithelial lymphocytes.     

Diagnostic criteria for malignancy in bile cytology and its usefulness

Fifty three bile specimens from 42 patients were reviewed to assess the diagnostic role of the bile cytology and to define more reliable cytologic indicators of malignancy. Forty three bile specimens came from 34 patients with malignant biliary strictures and 10 bile specimens were from eight patients with benign conditions. There were no false positives. The diagnostic specificity of bile cytology was 100% while diagnostic sensitivity was 55.8%. Overall diagnostic accuracy was 64.2%. We identified four key criteria as cytologic indicators of malignancy among 20 variables by using multiple regression analysis: loss of honeycomb arrangement, hyperchromatism, increased N/C ratio, and coarse chromatin. When bile specimens with three or more of these four criteria are thought to represent malignancy, the sensitivity of diagnosis of malignancy was 65.2%, specificity was 90% and diagnostic accuracy was 69.8%.     

99mTc-MIBI scan in mammary Pagets disease: a case report

Technetium-99m methoxyisobutylisonitrile (99mTc-MIBI) uptake is known to be increased in breast cancer because of increased blood flow from angiogenesis and heightened metabolism. We performed a 99mTc-MIBI scan in a patient with mammary Paget's disease. The patient had underlying invasive cancer in the same side of the breast. 99mTc-MIBI scan exhibited a scintigraphic image of the uptake from the invasive cancer lesion located deeply in the breast toward the epidermis. 99mTc-MIBI showed an uptake in the deeply located invasive cancer lesion as well as nipple lesion. Especially, the delayed phase of Tc-MIBI scan demonstrated the tumor site more accurately. In conclusion, 99mTc-MIBI scan could be a useful adjunct to clinical decision making in the management of Paget's disease of the breast.