Epiplakin modifies the motility of the HeLa cells and accumulates at the outer surfaces of 3‐D cell clusters

Elimination of epiplakin (EPPK) by gene targeting in mice results in acceleration of keratinocyte migration during wound healing, suggesting that epithelial cellular EPPK may be important for the regulation of cellular motility. To study the function of EPPK, we developed EPPK knock‐down (KD) and EPPK‐overexpressing HeLa cells and analyzed cellular phenotypes and motility by fluorescence/differential interference contrast time‐lapse microscopy and immunolocalization of actin and vimentin. Cellular motility of EPPK‐KD cells was significantly elevated, but that of EPPK‐overexpressing cells was obviously depressed. Many spike‐like projections were observed on EPPK‐KD cells, with fewer such structures on overexpressing cells. By contrast, in EPPK‐KD cells, expression of E‐cadherin was unchanged but vimentin fibers were thinner and sparser than in controls, and they were more concentrated at the peri‐nucleus, as observed in migrating keratinocytes at wound edges in EPPK^?/? mice. In Matrigel 3‐D cultures, EPPK co‐localized on the outer surface of cell clusters with zonula occludens‐1 (ZO‐1), a marker of tight junctions. Our results suggest that EPPK is associated with the machinery for cellular motility and contributes to tissue architecture via the rearrangement of intermediate filaments.     

LASER THERAPY FOR GLAUCOMA

Three laser techniques for the treatment of glaucoma are described. The short-burn technique of argon laser iridotomy permitted perforation of the peripheral iris in one session without serious complications, thereby alleviating pupillary block in angle-closure glaucoma. The rates of successful intraocular pressure (IOP) control and visual acuity loss due to cataract progression were similar in laser iridotomy and surgical iridectomy. The tonographic outflow facility remained unchanged after argon laser iridotomy.
Techniques of argon laser trabeculoplasty (AL TJ to the anterior and posterior aspects of the trabecular band were evaluated. The anterior ALT gave fewer complications, but a lower IOP control rate than the posterior ALT. Posterior ALT over 180° with about 50 burns gave fewer complications than treatment over 360° with about 100 burns, yet the final effects in IOP control were better after the 180° than after the 360° treatment.
Nd-YAG laser irradiation can separate dysgenetic iris insertion from the trabecular band in developmental glaucoma without serious complications. This Nd-YAG laser goniotomy may be useful for the treatment of juvenile primary developmental glaucoma.     

Comparison of altered signal intensity, position, and morphology of the TMJ disc in MR images corrected for variations in surface coil sensitivity.

OBJECTIVE: The purpose of this study is to evaluate the corrections of signal intensity of the temporomandibular joint (TMJ) disc caused by variations in sensitivity of the magnetic resonance imaging (MRI) surface coil, to compare the modified signal intensities of the posterior and anterior bands, and then to evaluate the relationship of the signal intensity difference to altered disc position and morphology in a group of TMJ patients. STUDY DESIGN: MRI was performed on 96 joints. All patients underwent imaging in axial, coronal, and sagittal planes using fast-spin echo sequences (FSE). The images were taken in the closed, partially opened, and maximum opened mouth positions in 2 sequences. Classifications were made according to the position and morphology of the disc. TMJs were divided into normal, anterior disc displacement with reduction (ADDwR), anterior disc displacement without reduction (ADDwoR), and partial anterior disc displacement with reduction (PDDwR). Disc morphology was subdivided as biconcave, lengthened, biconvex, thick posterior band, and others (defined as folded and rounded). The correction of the inhomogeneous sensitivity of the surface coil was done with the original software. The signal intensities (SI) of the posterior band and anterior band of TMJ discs were measured. The correlations among the groups of TMJs and disc morphologies and SI were statistically analyzed by using Bonferroni/Dunn multicomparison method test. RESULTS: Of the total number of joints studied with the help of MRI, 37 were normal, 12 exhibited ADDwR, 32 ADDwoR, and 9 PDDwR. The corrected MR images indicated that SI of the posterior bands were higher than the anterior band of the discs. It can also be concluded that the SI of the posterior bands increased significantly in the following order: normal, PDDwR, ADDwR, and ADDwoR, while there is no statistical difference in the SI of the anterior band of the discs. In ADDwR and ADDwoR, thick posterior band is the most common shape. In normal TMJ, the biconcave shape is identified as the most frequently encountered shape. CONCLUSIONS: It was demonstrated that the SI of the posterior bands increase with the progress of internal derangement, and was found to be higher than that of the anterior band of the discs. It appears that disc degeneration starts from the posterior band of the disc.     

Brachytherapy for early oral tongue cancer: low dose rate to high dose rate

To examine the compatibility of low dose rate (LDR) with high dose rate (HDR) brachytherapy, we reviewed 399 patients with early oral tongue cancer (T1-2N0M0) treated solely by brachytherapy at Osaka University Hospital between 1967 and 1999. For patients in the LDR group (n = 341), the treatment sources consisted of Ir-192 pin for 227 patients (1973-1996; irradiated dose, 61-85 Gy; median, 70 Gy), Ra-226 needle for 113 patients (1967-1986; 55-93 Gy; median, 70 Gy). Ra-226 and Ir-192 were combined for one patient. Ir-192 HDR (microSelectron-HDR) was used for 58 patients in the HDR group (1991-present; 48-60 Gy; median, 60 Gy). LDR implantations were performed via oral and HDR via a submental/submandibular approach. The dose rates at the reference point for the LDR group were 0.30 to 0.8 Gy/h, and for the HDR group 1.0 to 3.4 Gy/min. The patients in the HDR group received a total dose of 48-60 Gy (8-10 fractions) during one week. Two fractions were administered per day (at least a 6-h interval). The 3- and 5-year local control rates for patients in the LDR group were 85% and 80%, respectively, and those in the HDR group were both 84%. HDR brachytherapy showed the same lymph-node control rate as did LDR brachytherapy (67% at 5 years). HDR brachytherapy achieved the same locoregional result as did LDR brachytherapy. A converting factor of 0.86 is applicable for HDR in the treatment of early oral tongue cancer.     

A preliminary and clinical study of radiation therapy for tongue carcinoma

Four hundred and seventy-four patients with squamous cell carcinomas of the tongue were treated with radiation therapy at the Department of Radiology, Osaka University Hospital between 1978 and 1987. Brachytherapy was mainly employed for the majority of the patients with T1 and T2 lesions, whereas the combination of external irradiation and brachytherapy was the treatment of choice for the patients with infiltrative T2 and more advanced lesions. Patients treated with brachytherapy alone were given an estimated tumor dose of about 70Gy within a week. Patients treated with the combined therapy received an estimated tumor dose of about 30Gy to 50Gy within 3 to 5 weeks from external irradiation, followed by 50Gy to 60Gy within a week from brachytherapy. The treatments used in this study improved the survival. The 2-year local control rates were 85% in T1, 77% in T2, 60% in T3 and 33% in T4 lesions. The 5-year actuarial survival rates according to the T stage were 78% in T1, 64% in T2, 50% in T3 and 40% in T4 lesions. One hundred and four patients(28%) of three hundred and sixty nine patients who had negative lymph nodes clinically had cervical lymph node metastases subsequently. Whereas twenty six patients(25%) of one hundred and five patients who had positive lymph nodes clinically proved to be false positive. The incidence of osteoradionecrosis was 6.0% in patients treated with the combination of external irradiation(30Gy) and brachytherapy(60Gy), while it was 1.7% in patients treated with brachtherapy alone(70Gy). Brachytherapy is a fairly good therapeutic technique for the majority of the patients with tongue cancers. The multicombined treatment modality could provide the chance for cure of advanced lesions.     

Effects of cepharanthine with or without Z-100 against leukopenia during radiation therapy for oral cancer

Objectives

The purpose of this study was to evaluate the clinical influence of the administration of cepharanthine with or without Z-100 on the number of leukocytes during external beam radiation therapy (EBRT) in patients with oral cancer.

Methods

Data from 43 patients with oral cancer who received EBRT were evaluated retrospectively. During EBRT, 14 patients received neither cepharanthine nor Z-100 (group A), 19 patients were administered 6?mg/day peroral cepharanthine (group B), and 10 patients were administered both 6?mg/day peroral cepharanthine and Z-100 injections (group C). Blood tests were performed for all patients before, during, and after EBRT. The maintenance rates of white blood cells, lymphocytes, and neutrophils were evaluated.

Results

The average maintenance rates of white blood cells in group A were 84.7?% during the first half of EBRT, 81.5?% during the second half of EBRT, and 88.2?% at 1?month after EBRT. The corresponding rates were 112.6, 98.7, and 101.0?% in group B, and 97.1, 101.5, and 90.8?% in group C, respectively. The average maintenance rate of white blood cells in group B was significantly higher than that in group A during the first half of EBRT (P?<?0.05).

Conclusions

Administration of cepharanthine may be useful for preventing leukopenia in patients with oral cancer treated by EBRT.     

3' Phosphatase activity toward phosphatidylinositol 3,4-bisphosphate [PI(3,4)P2] by voltage-sensing phosphatase (VSP)

Voltage-sensing phosphatases (VSPs) consist of a voltage-sensor domain and a cytoplasmic region with remarkable sequence similarity to phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a tumor suppressor phosphatase. VSPs dephosphorylate the 5' position of the inositol ring of both phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P(3)] and phosphatidylinositol 4,5-bisphosphate [PI(4,5)P(2)] upon voltage depolarization. However, it is unclear whether VSPs also have 3' phosphatase activity. To gain insights into this question, we performed in vitro assays of phosphatase activities of Ciona intestinalis VSP (Ci-VSP) and transmembrane phosphatase with tensin homology (TPTE) and PTEN homologous inositol lipid phosphatase (TPIP; one human ortholog of VSP) with radiolabeled PI(3,4,5)P(3). TLC assay showed that the 3' phosphate of PI(3,4,5)P(3) was not dephosphorylated, whereas that of phosphatidylinositol 3,4-bisphosphate [PI(3,4)P(2)] was removed by VSPs. Monitoring of PI(3,4)P(2) levels with the pleckstrin homology (PH) domain from tandem PH domain-containing protein (TAPP1) fused with GFP (PH(TAPP1)-GFP) by confocal microscopy in amphibian oocytes showed an increase of fluorescence intensity during depolarization to 0 mV, consistent with 5' phosphatase activity of VSP toward PI(3,4,5)P(3). However, depolarization to 60 mV showed a transient increase of GFP fluorescence followed by a decrease, indicating that, after PI(3,4,5)P(3) is dephosphorylated at the 5' position, PI(3,4)P(2) is then dephosphorylated at the 3' position. These results suggest that substrate specificity of the VSP changes with membrane potential.