Effects of immunosuppressants on induction of regulatory cells after intratracheal delivery of alloantigen

BACKGROUND: We previously reported that intratracheal delivery (ITD) of alloantigen generated regulatory cells in mice. Here, we examined the effect of various doses of conventional immunosuppressants (FK506, cyclosporine A, azathioprine, mycophenolate mofetil, and rapamycin) on inducing regulatory cells in our model. METHODS: CBA mice (primary recipients) were given C57BL/6 splenocytes by ITD and either no additional treatment or various doses of an immunosuppressant. Seven days later, splenocytes from these mice were adoptively transferred into naive secondary CBA recipients that underwent C57BL/6 cardiac grafting the same day. RESULTS: Adoptive transfer from primary recipients given ITD of splenocytes alone induced prolonged allograft survival in secondary recipients (median survival time [MST], 50 days), suggesting that regulatory cells were generated. When ITD of alloantigen was combined with daily administration of 0.1 mg/kg FK506 or 0.2 mg/kg rapamycin, graft survival was similarly prolonged (MST 55 and 50 days, respectively). When combined with 20 or 40 mg/kg MMF or 0.4 mg/kg rapamycin, the majority of recipients demonstrated indefinite survival (MST, >100 days in all groups). When ITD of alloantigen was combined with 0.3, 0.5, or 1.0 mg/kg FK506; 5, 10, or 25 mg/kg cyclosporine A; or 1.0 or 2.0 mg/kg azathioprine, allografts were rejected acutely (MST 7-13 days). CONCLUSION: Generation of regulatory cells by ITD of alloantigen was facilitated by mycophenolate mofetil and high doses of rapamycin but abrogated by cyclosporine A, azathioprine, and high doses of FK506. Low doses of rapamycin and of FK506 did not interfere with generation of regulatory cells.     

Prestin-dependent and prestin-independent motility of guinea pig outer hair cells

The motile response of isolated guinea pig outer hair cells (OHCs) was investigated using a combination of whole-cell patch clamp recording and continuous video image analysis. OHC's length, width, and area were measured from video images and the cell volume estimated from these values. Morphological data was then correlated with electrophysiological recordings of whole-cell current, membrane potential and voltage-dependent non-linear capacitance. Electromotility was evoked either by manipulating the membrane potential under voltage-clamp conditions or by exposing OHCs to high K+ solutions. Other motile responses were investigated in voltage-clamp experiments at constant holding potential, or exposing OHCs to solutions that did not affect the membrane potential. We found that electrical stimulation evoked voltage-dependent changes in OHC's length, width and area but not in cell volume regardless of the time course of stimulation. Moreover, changes in cell area were always associated with both voltage-dependent motility and non-linear capacitance, suggesting prestin dependency. In contrast, voltage-independent motile responses at constant membrane potential, which are presumed to be prestin-independent, were associated with changes in cell length, width and volume without significant changes in area. Area measurements, then, become a tool to investigate the simultaneous occurrence of both prestin-dependent and prestin-independent OHC motilities, and for evaluating the individual contribution of each mechanism to the total cell movement.     

Reduction of radiation dose at HRCT of the temporal bone in children

PURPOSE: The purpose of this study was to reduce the radiation exposure of the eye lens in high resolution computed tomography (HRCT) of the temporal bone using an experimental phantom. MATERIALS AND METHODS: The HRCT image that was used for analysis was obtained by changing parameters including effective-mAs (E-mAs), distance coverage, and height of object in the Y-axis. Radiation exposure was measured to calculate equivalent doses by glass rod dosimeters that were fixed above the right orbit parallel to the body axis. Deterioration in image quality was evaluated by three radiologists and the following three-point rating method was employed: grade 1 (good image quality without diagnostic limitations), grade 2 (image was deteriorated, but there were no diagnostic limitations), and grade 3 (image was deteriorated with diagnostic limitations). RESULTS: Assuming that the equivalent dose was y (mSv), and E-mAs was x, a simple regression line, y=0.506x-0.494 (decision coefficient, R2=0.999), was obtained. A standard deviation (S.D.) less than 120 (E-mAs, 220-120) was judged as grade 1, an S.D. between 120 and 150 was judged as grade 2, and an S.D. higher than 150 was judged as grade 3, indicating that deterioration of the quality of images with reduced E-mAs affected the diagnosis by imaging at S.D. higher than 150. CONCLUSION: Radiation dose at the eye lens in HRCT could be reduced up to an equivalent dose corresponding to 70 mAs without compromising diagnostic quality in the phantom experiment.     

The impact of angiogenesis on the prognosis of advanced renal cell carcinomas

We studied the relationship between angiogenic factors and clinical responses in advanced renal cell carcinomas (RCCs) and evaluated the angiogenic factors to clarify the potential impact of these factors on the cancer-specific survival. From January 1990 to December 2000, 148 patients underwent a nephrectomy for RCCs at our institution. Of the 32 patients who had distant metastasis, 17 met the histopathologic analysis requirements for an immuno-histochemical investigation. Fifteen of them were administered interferon-gamma and the remaining two patients were added to interferon-alpha and eight of seventeen patients also underwent radiation therapy. Both thymidine phosphorylase (TP) and Factor VIII immunostaining were performed. The overall survival rates at 1, 5 and 10 years were 82.4%, 30% and 30%, respectively. Three of these patients were diagnosed with lung metastasis and a complete response was seen in two, while a partial response was observed in one. In addition another patient who was diagnosed with bone metastasis also showed a partial response (group A). The remaining 13 patients showed progressive disease (group B). Group A had a higher TP-positive ratio (TP-PR) than that of group B. A multivariate analysis of the clinicopathologic data showed that a positive mean vascular area (PMVA) could be an independent factor regarding the potential impact of these factors on a long survival in advanced RCCS. PMVA was thus found to be an independent factor regarding the prognosis with advanced RCCs.