Confocal microscopic analysis of the interaction between cisplatin and the copper transporter ATP7B in human ovarian carcinoma cells.

Some cisplatin (DDP)-resistant cells overexpress the copper export transporter ATP7B, and cells molecularly engineered to overexpress ATP7B are resistant to DDP. The interaction of Cu with ATP7B normally triggers its relocalization from the perinuclear region to more peripheral vesicles. To investigate the interaction of DDP with ATP7B, we examined the effect of DDP on the subcellular localization of ATP7B using human ovarian carcinoma cells expressing a cyan fluorescent protein (ECFP)-tagged ATP7B (2008/ECFP-ATP7B). ATP7B expression was confirmed in 2008/ECFP-ATP7B cells by Western blotting, and its functionality was documented by showing that it rendered the cells 1.9-fold resistant to CuSO(4) and 4.1-fold resistant to DDP and also reduced the accumulation of both drugs. There was greater sequestration of Pt into intracellular vesicles in the 2008/ECFP-ATP7B cells than in the 2008/ECFP cells. Confocal digital microscopy revealed that ECFP-ATP7B localized in the perinuclear region in absence of drug exposure and that both Cu and DDP triggered relocalization to more peripheral vesicular structures. A fluorescein-labeled form of DDP that retained cytotoxicity and was subject to the same mechanisms of resistance as DDP colocalized with ECFP-ATP7B in the 2008/ECFP-ATP7B cells, whereas the same fluorochrome lacking the DDP moiety did not. These results provide evidence that DDP directly interacts with ATP7B to trigger its relocalization and that ATP7B mediates resistance to DDP by sequestering it into vesicles of the secretory pathway for export from the cell.     

Septate versus bicornuate uteri: errors in imaging diagnosis

Since two müllerian defects, the septate and bicornuate uteri, are no longer repaired by means of the same operative approach, an accurate preoperative diagnosis of these anomalies is now critical. A septum can be removed by means of hysteroscopic metroplasty. However, repair of a bicornuate uterus still requires abdominal surgery. Hysterosalpingography (HSG) has been the primary diagnostic modality for müllerian defects. On the basis of 63 patients, HSG findings alone, as interpreted by the radiologist, had a diagnostic accuracy of 55%. When this was supplemented with a gynecologic evaluation, the diagnostic accuracy improved to only 62.5%. However, when a diagnostic protocol that include ultrasound (US) examination with HSG was used for evaluating müllerian defects, the diagnostic accuracy improved to 90%, with all errors being noncritical. Therefore, it is concluded that HSG alone is not adequate to make the distinction between a septate and a bicornuate uterus unless the angle of divergence of two straight uterine cavities is 75 degrees or less, indicating a septate uterus. Luteal-phase US is frequently necessary to distinguish between these anomalies or to diagnose them in combination.     

Residual mediastinal masses in Hodgkin disease: prediction of size with MR imaging

Eighteen patients with mediastinal involvement of Hodgkin disease were examined with magnetic resonance (MR) imaging before and during therapy to find out if size of residual masses could be predicted from the MR characteristics of the tumor at diagnosis. After the first treatment, a significant decrease in T2 values and signal intensity ratios of tumor to fat and tumor to muscle was found in all patients. There was no significant change in T1 values. The relative decrease in tumor size correlated well with signal intensity ratios and poorly with T2 values of the original tumor. No correlation with T1 values was found. The authors conclude that size of the residual mass can be predicted from the initial size of the tumor and the signal intensity ratios at diagnosis. Since the degree of low signal intensity in the tumor before treatment probably reflects the amount of fibrotic tissue, these results support the hypothesis that residual masses after treatment are remnants of the fibrotic stroma of the original tumor.     

Novel thiosemicarbazones derived from formyl- and acyldiazines: synthesis, effects on cell proliferation, and synergism with antiviral agents.

The synthesis of a series of novel thiosemicarbazones (TSC's) derived from various alkyl diazinyl (3-pyridazinyl, 4-pyrimidinyl, 2-pyrazinyl) ketones and 3-pyridazinecarbaldehyde and their evaluation against herpes simplex virus (HSV) and human immunodeficiency virus (HIV) as well as the determination of their cytotoxicity are described. In addition, the effects of combination of such TSC's with the well-known antiviral drugs acyclovir (ACV) and 3'-azido-3'-deoxythymidine (AZT) were studied. Under our experimental conditions, i.e. determination of virus-induced cytopathic effect upon infection of HUT78 cells with HSV-1 and upon infection of MT4 cells with HIV-1, no antiviral activity could be detected with any of the TSC's. However, pronounced effects on proliferation of these rapidly growing T4 lymphocyte cell lines were observed. Clear structure-activity relationships with regard to these cytotoxic effects could be established: compared to pyridine, pyrazine, or pyrimidine-derived TSC's most of the 3-pyridazinyl congeners investigated are less cytotoxic; introduction of a methyl group into C-6 of the pyridazine system or prolongation of the acyl moiety in these compounds has essentially no influence; all compounds bearing an N,N-dimethylamino or a cycloamino substituent are much more toxic than those with an NH2 or NHR substituent; the nature of R in the latter type of compounds has only moderate influence. It has been reported that combination of TSC's with the antiviral agent acyclovir (ACV) results in potentiation of this well-known drug. We evaluated the potential of our series of novel TSC's in combination with ACV for inhibition of HSV-1-induced cytopathic effect in HUT78 cells and in combination with 3'-azido-3'-deoxythymidine (AZT) for inhibition of HIV-1-induced cytopathic effect in MT4 cells. Only four compounds out of this series, all characterized by an unsubstituted NH2 group, exhibited moderate synergism with the above mentioned antiviral drugs. Our results do not support the previously expressed opinion that TSC's are selective antiviral agents. In our test systems no evidence for inhibition of virus-induced cytopathic effect was obtained. The TSC derivatives exhibited a broad range of cytotoxic effects, some at concentrations considerably below those reported to have antiviral efficacy. Several of our novel diazine-derived compounds proved advantageous over the previously described pyridine analogues with regard to cytotoxicity. Moderate synergism could be detected for relatively noncytotoxic TSC's with the antiviral drugs ACV (antiherpes) and AZT (anti-HIV).     

Radiation induced carcinoma of the temporal bone

This report presents a case of squamous cell carcinoma of the temporal bone induced by high dose radiation therapy. Several important surgical techniques are stressed which serve to make this procedure far safer in the avoidance of catastrophic hemorrhage.