Local Control of Squamous Cell Carcinoma of the Cervix Treated with CT-based Three-dimensional Image-Guided Brachytherapy with or without Central Shielding.

The purposes of this retrospective study were to analyze local control of squamous cell carcinoma of the cervix treated with computed tomography (CT)-based image-guided brachytherapy (IGBT), as well as the factors affecting local control. A total of 39 patients were analyzed. The prescribed dose to the pelvis was 45-50 Gy with or without central shielding (CS). IGBT was delivered in 1-5 fractions. The total dose for high-risk clinical target volume (HR-CTV) was calculated as the biologically equivalent dose in 2-Gy fractions. The median follow-up period was 29.3 months. The 2-year overall survival and local control rates were 97% and 91%, respectively. In univariate analysis, the dose covering 90% of the HR-CTV (D90) and tumor size were found to be significant factors for local control. The cutoff values of tumor size and D90 for local control were 4.3 cm (area under the curve [AUC] 0.75) and 67.7 Gy (AUC 0.84) in the CS group and 5.3 cm (AUC 0.75) and 73.7 Gy (AUC 0.78) in the group without CS, respectively. However, though the local control of CT-based IGBT was favorable, the results suggested that the dose required for tumor control may differ depending on the presence of CS.

Simultaneous Integrated Boost Volumetric Modulated Arc Therapy for Middle or Lower Esophageal Cancer Using Elective Nodal Irradiation: Comparison with 3D Conformal Radiotherapy.

We investigated the feasibility of simultaneous integrated boost (SIB) volumetric modulated arc therapy (VMAT) using elective nodal irradiation (ENI) for middle or lower esophageal cancer and compared it with three-dimensional conformal radiotherapy (3D-CRT). The study included 15 patients. The prescribed doses included a standard dose (50.4 Gy) and a high dose (60 Gy) for the planning target volume (PTV) of the involved lesions. The objective of the whole lung volume receiving ≥ 20 Gy (V20Gy) was < 30%, and the mean lung dose (MLD) was < 20 Gy. The volumes of the lung receiving 5 Gy (V5Gy) and the heart receiving 30-50 Gy (V30-50Gy) were kept as low as reasonably achievable. As a result, SIB-VMAT showed superior dose conformity for the PTV (p<0.001). Although the lung V5Gy was significantly increased (p<0.001), the V20Gy and MLD showed no significant increase. The heart V30-50Gy showed a > 20% reduction in the mean against 3D-CRTs. Our results demonstrate the feasibility of SIB-VMAT for the treatment of middle or lower esophageal cancer with ENI. Although attention should be paid to the low-dose area of the lungs, SIB-VMAT would be a promising treatment option with improved outcomes for esophageal cancer.

An Adult Case of Nasal Chondromesenchymal Hamartoma: Imaging Characteristics Including Diffusion-Weighted Images.

Nasal chondromesenchymal hamartoma (NCMH), a rare, benign, nasal cavity tumor, typically occurs in children. Differential diagnosis is difficult because NCMH often presents with non-specific findings, including cystic components and invasion of the surrounding area on T2-weighted magnetic resonance images. Here, we present a rare adult case of NCMH, with no clear hyperintensity on diffusion-weighted images (DWI), and bone remodeling on the tumor margins on computed tomography. To the best of our knowledge, this is the first report of DWI on NCMH, and these findings, which suggest benign disease, may be useful in diagnosing NCMH.

Feasibility of 5-mm vs 2.5-mm width multileaf collimator in noncoplanar volumetric modulated arc stereotactic radiotherapy for multiple brain metastases.

The aim of this study was to examine the feasibility of noncoplanar volumetric modulated arc stereotactic radiotherapy (VMAT-SRT) using a 5-mm multileaf collimator (MLC) for multiple brain metastases. We identified 34 multiple-target cases (3 to 19 targets in each case) with a total of 257 of targets and constructed noncoplanar VMAT-SRT plans using 5-mm and 2.5-mm MLCs with 4-arc. The prescribed dose was 36 Gy/6 fr. Plans were evaluated using the Paddick conformity indices (PCI), Paddick gradient index (PGI), and normal brain dose (NBD, equal to the mean brain dose minus gross tumor volume). There were no significant differences in PCI (median [range]: 5 mm, 0.88 [0.78 to 0.94]; 2.5 mm, 0.89 [0.78 to 0.94]; p= 0.691), PGI (median [range]: 5 mm, 3.96 [2.21 to 6.63]; 2.5 mm, 3.96 [2.24 to 6.45]; p= 0.358), or NBD (median [range]: 5 mm, 7.5 Gy [2.5 to 12.4]; 2.5 mm, 7.5 Gy [2.5 to 12.5]; p= 0.675). The performance of the 5-mm MLC was not inferior to the 2.5-mm MLC in applications of noncoplanar VMAT-SRT for multiple brain metastases with regards to dose conformity, gradient, and NBD. This study provides the necessary background for generalizing noncoplanar VMAT-SRT approaches in treating multiple brain lesions.

Preliminary retrospective investigation of FDG-PET/CT timing in follow-up of ablated lung tumor.

OBJECTIVE: The aim of this study was to clarify the most appropriate follow-up initiation time point for positron emission tomography (PET)/computed tomography (CT) following radio frequency ablation (RFA) of lung tumors, and the cutoff values of maximum standard uptake value (SUV(max)) to evaluate local tumor progression. METHODS: We enrolled 15 patients (8 men, median age 62 years) with 60 tumors, who were treated with RFA of lung tumors and underwent fluorodeoxyglucose (FDG)-PET/CT following RFA. Local tumor progression was assessed by periodic chest CT images prior to and following intravenous administration of a contrast medium. The SUV(max) of three periods, namely, 0-3 months, 3-6 months, and 6-9 months after RFA, was evaluated. The appropriate time point for follow-up initiation and the cutoff value of SUV(max) were determined using receiver-operating characteristic (ROC) analysis. RESULTS: The median follow-up period was 357 days. Of 60 tumors, 10 showed local progression. The area under the ROC curve (Az) for the 6-9 months (P = 0.044) was the largest and almost equal to that of the 3-6 months (P = 0.024). Az for the 0-3 months was the smallest and statistically insignificant (P = 0.705). The cutoff value of 1.5 of SUV(max) at 3-9 months after RFA showed 77.8% sensitivity and 85.7-90.5% specificity. CONCLUSIONS: The appropriate follow-up initiation time point is at least 3 months following RFA. Thus, SUV(max) is a useful and reliable predictive indicator.

Assessment of mean transit time in the engrafted lung with 133Xe lung ventilation scintigraphy improves diagnosis of bronchiolitis obliterans syndrome in living-donor lobar lung transplant recipients.

OBJECTIVE: Staging of bronchiolitis obliterans syndrome (BOS) following lung transplantation is based on declines in forced expiratory volume in 1 s (FEV(1)). The aim of this study was to evaluate the usefulness of (133)Xe ventilation scintigraphy in the early detection of BOS following living-donor lobar lung transplantation (LDLLT), to compare (133)Xe washout imaging with computed tomography (CT) findings for early detection of BOS following LDLLT, and to evaluate (133)Xe washout imaging by quantitative analyses. METHODS: Subjects comprised 30 double-lung recipients and 1 single-lung recipient, who had undergone LDLLT at our institution and survived more than 1 year. Clinically diagnosed BOS developed in six recipients. Declines in graft function were evaluated using a combination of three methods, namely, dynamic spirometry, high-resolution CT (HRCT), and (133)Xe ventilation scintigraphy. Findings for all transplanted lungs were compared between CT and (133)Xe washout imaging. (133)Xe washout imaging was assessed using mean transit time (MTT) of bi-and unilateral lungs. Correlations between MTT of bilateral lungs and FEV(1)% were evaluated. Differences in MTT between BOS and non-BOS lungs, and between non-BOS and donor lungs were also evaluated on unilateral lungs. Appropriate cut-off values of MTT of unilateral lungs were set for the diagnosis of BOS. RESULTS: In all six BOS cases, prolonged-washout images of engrafted lungs revealed early-phase BOS with declines from baseline FEV(1), whereas only one BOS case could be detected using early CT findings of BO (bronchodilatation, decrease in number and size of pulmonary vessels, thickening of septal lines, and volume reduction). A significant correlation was identified between MTT and FEV(1)% (r = -0.346, P < 0.0001). MTT of unilateral lungs was significantly longer in BOS lungs than in non-BOS lungs (P < 0.0001). The cut-off MTT of unilateral lungs for the diagnosis of BOS was set at 64.77 s. CONCLUSIONS: Our data show that (133)Xe washout imaging offers excellent potential for early detection of BOS compared with early CT findings. Using (133)Xe washout imaging and MTT with radioactive tracer offers a noninvasive indication of selective ventilatory function in engrafted lungs following LDLLT. MTT appears useful for identifying BOS following LDLLT and allows quantitative evaluation of graft function in unilateral lungs.

Plan quality comparison between 4-arc and 6-arc noncoplanar volumetric modulated arc stereotactic radiotherapy for the treatment of multiple brain metastases.

To compare the plans of 4-arc and 6-arc noncoplanar volumetric modulated arc stereotactic radiotherapy (VMA-SRT) for multiple brain metastases and to investigate the cutoff value for the tumor number and volume for 6-arc rather than 4-arc VMA-SRT. We identified 24 consecutive multiple-target cases (3 to 19 targets in each case) with 189 total targets. We constructed plans using both 4- and 6-arc noncoplanar VMA-SRT. The prescribed dose was 36 Gy/6 fr, and it was delivered to 95% of the planning target volume (PTV). The plans were evaluated for the dose conformity using the Radiation Therapy Oncology Group and Paddick conformity indices (RCI and PCI), fall-off (Paddick gradient index [PGI]), and the normal brain dose. The median (range) RCI, PCI, and PGI was 0.94 (0.92 to 0.99), 0.89 (0.77 to 0.94), and 3.75 (2.24 to 6.54) for the 4-arc plan and 0.94 (0.91 to 0.98), 0.89 (0.76 to 0.94), and 3.65 (2.24 to 6.5) for the 6-arc plan, respectively. The median (range) of the normal brain dose was 910.3 cGy (381.4 to 1268.9) for the 4-arc plan and 898.8 cGy (377 to 1252.9) for the 6-arc plan. The PGI of the 6-arc plan was significantly superior to that of the 4-arc plan (p = 0.0076), and the optimal cutoff values for the tumor number and volume indicative of 6-arc (and not 4-arc) VMA-SRT were cases with ≥ 5 metastases and a PTV of ≥ 12.9 mL, respectively. The PCI values, however, showed no significant difference between the 2 plans. We believe these results will help in considering the use of 6-arc VMA-SRT for multiple brain metastases.

Evaluation of esophageal varices by multidetector-row CT: correlation with endoscopic 'red color sign'.

To evaluate the ability of multidetector-row CT (MDCT) to predict a risk of hemorrhage in patients with esophageal varices, a total of 40 MDCT scans were performed in 29 patients who had been diagnosed with esophageal varices by conventional upper gastrointestinal tract endoscopy. In 11 patients, MDCT was performed both before and after endoscopic injection sclerotherapy (EIS). Endoscopically, the red color sign (RC sign) was present in 28 scans. Of the 11 patients who underwent EIS, the RC sign disappeared after EIS in 9. The MDCT scans were obtained in the arterial, portal, and equilibrial phases, and the portal phase images were used in this study. Subsequently, the extent of esophageal varices was categorized into four MDCT scores. The variceal score, the maximum short axis of the varices, and the presence of palisade vein dilatation obtained from MDCT had significant correlation with endoscopic variceal forms, and the presence and severity of RC sign, respectively (p<0.01). All cases with a maximum minor axis of more than 4 mm showed positive RC sign. MDCT was useful in the evaluation of esophageal varices for predicting a risk of hemorrhage.

Computed tomography findings of congenital generalized lipodystrophy: multiple nodular fatty liver and diffuse sclerosis of bones.

Congenital generalized lipodystrophy (CGL) is a rare autosomal recessive disease that is also referred to as Berardinelli-Seip syndrome. It is characterized by a lack of adipose tissue throughout the body from birth, muscular hypertrophy, advanced bone age, fatty liver, and insulin resistance. We describe computed tomography (CT) and magnetic resonance findings for a 35-year-old woman with CGL. Multiple nodular, well-defined regions of fatty infiltration of the liver are rare and have never been previously reported in a patient with lipodystrophy. To our knowledge, this is the first report describing CT findings of bone sclerotic changes associated with CGL.

F-18 FDG PET demonstration of a thyroid metastasis in a patient with colon cancer.

A 51-year-old man with a history of surgical removal of sigmoid colon cancer underwent F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) to search for distant metastases and/or local recurrence because the carcinoembryonic antigen level was elevated. F-18 FDG PET images showed increased focal FDG uptake in the left lobe of the thyroid. Computed tomography images showed thyroid tumor in the left lobe as well as F-18 FDG PET images. Thereafter he underwent thyroidectomy and the resected specimen was histopathlogically shown to have thyroid metastasis from colon cancer. F-18 FDG PET was useful to detect thyroid metastasis from colon cancer as well as the most frequently seen metastatic sites such as liver, lungs, and lymph nodes.

Positive gallium-67 and thallium-201 scans in thymic rebound after chemotherapy for lymphoma.

It is a diagnostic problem to distinguish thymic rebound or rebound thymic hyperplasia from thymic malignancy, but it is frequently made more difficult because most patients have had previous malignancies. Recently we evaluated a six-year-old girl with thymic rebound after chemotherapy for lymphoma, by both gallium-67 and thallium-201 scans. On gallium-67 scan, intensive uptake was seen in the anterior mediastinum. CT revealed a triangular-shaped, homogeneous mass in the anterior mediastinum. On early scan of thallium-201 study, slight accumulation was seen in the anterior mediastinum and was enhanced in delayed scans. Considering the clinical state and imaging results, thymic rebound after chemotherapy was the most likely diagnosis, and follow-up observation was done without therapy. During the course, there were no signs of relapse. Some reports have described both positive and negative thallium-201 accumulation in thymic rebound. Although more experience with similar cases is necessary, it is likely that thallium-201 also tends to accumulate in thymic rebound as well as gallium-67.

Cerebral perfusion MR imaging using FAIR-HASTE in chronic carotid occlusive disease: comparison with dynamic susceptibility contrast-perfusion MR imaging.

To determine the efficacy of flow-sensitive alternating inversion recovery using half-Fourier single-shot turbo spin-echo (FAIR-HASTE) in detecting cerebral hypoperfusion in chronic carotid occlusive disease, we subjected 12 patients with various degrees of cervical internal carotid artery stenoses and/or occlusion (Stenosis group) and 24 volunteers (Normal group) to FAIR-HASTE. In addition, 10 out of 12 patients in the Stenosis group underwent dynamic susceptibility contrast-perfusion magnetic resonance imaging (DSC-pMRI) before and after revascularization in the dominantly affected side. The absolute asymmetry indexes (AIs) of both cerebral hemispheres in the Normal and Stenosis groups were compared in FAIR-HASTE. In addition, the AIs were compared with those in the Stenosis group before and after revascularization in both FAIR-HASTE and regional cerebral blood flow (rCBF), calculated with DSC-pMRI. A statistically significant difference was recognized between the AIs in the Normal and Stenosis groups (AI = 2.25 +- 1.92, 8.09 +- 4.60, respectively ; p < 0.0001). Furthermore, in the Stenosis group the AIs on both FAIR-HASTE (8.88 +- 4.93, 2.22 +- 1.79, respectively ; p = 0.0003) and rCBF (7.13 +- 3.57, 1.25 +- 1.33, respectively ; p = 0.0003) significantly decreased after revascularization. In the Stenosis group, before revascularization, signal intensity on both FAIR-HASTE and rCBF had a tendency to be lower in the dominantly affected side. FAIR-HASTE imaging was useful in the detection and evaluation of cerebral hypoperfusion in chronic occlusive carotid disease.

Hydrogen peroxide overload increases adriamycin-induced apoptosis of SaOS(2)FM, a manganese superoxide dismutase-overexpressing human osteosarcoma cell line.

We previously developed a new microscopic observation system that enables time-lapse quantitative analysis of apoptosis and necrosis. With this system we quantitatively analyzed adriamycin (ADR)-induced cell death using manganese superoxide dismutase (MnSOD)- and wild-type p53-gene transfectants on SaOS(2), a p53-deficient human osteosarcoma cell line. A highly MnSOD-overexpressing cell line, SaOS(2)FM(H), acquired ADR-tolerance compared to the parent cell line SaOS(2). The ADR-tolerance of SaOS(2)FM(H) diminished by L-buthionine-[S,R]-sulfoximine (BSO), which did not change ADR-sensitivity of SaOS(2), to the similar ADR-sensitivity of SaOS(2). A wild-type p53-expressing cell line, SaOS(2)wtp53, significantly increased in ADR-sensitivity compared to SaOS(2). This ADR-sensitivity of SaOS(2)wtp53 was enhanced by BSO. When isosorbide 5-mononitrate was combined with BSO, isosorbide 5-mononitrate increased ADR sensitivity of a moderately MnSOD-overexpressing cell line, SaOS(2)FM(L), decreased that of SaOS(2) FM(H), and did not change those of SaOS(2) and SaOS(2)wtp53 compared to BSO alone. Time-lapse microscopic observations during ADR treatment for 24 h indicated that the most cells of each cell line underwent apoptosis, and a few cells (less than 11%) died by necrosis. When cells were treated with iso-concentration of ADR, apoptosis of SaOS(2)FM(H) was less than that of SaOS(2). BSO, which did not change ADR-sensitivity of SaOS(2), increased appearance rate of ADR-induced apoptosis, but not necrosis of MnSOD-overexpressing cell lines. When iso-survival dose of ADR, which reduced surviving fraction to 0.01, was given for each cell line, no difference was observed in appearance of either apoptosis or necrosis between SaOS(2) and MnSOD-overexpressing cell lines. On the other hands, appearance of both apoptosis and the following secondary necrosis of SaOS(2) wtp53 was significantly accelerated compared to those of SaOS(2). These findings indicate that hydrogen peroxide overload on p53-independent pathway due to MnSOD overexpression plus BSO might increase the apoptosis frequency without acceleration of apoptotic process of each cell, resulting in negating ADR-tolerance of MnSOD-overexpressing cell lines.

Manganese superoxide dismutase overexpression changes plating efficiency bidirectionally according to change in redox for SaOS2 human osteosarcoma cell line.

Manganese superoxide dismutase (MnSOD) is a mitochondrial enzyme that converts cytotoxic superoxide radicals into hydrogen peroxide. MnSOD activity is lower in tumor cells, and MnSOD overexpression reportedly ameliorates malignant phenotypes. We established stable MnSOD overexpressing cell lines from a human osteosarcoma cell line, SaOS2, and then investigated the effects of MnSOD overexpression on plating efficiency (PE) and the involvement of reactive oxygen species, including nitric oxide (NO) in those effects. The PE of SaOS2FM(L), a moderate MnSOD overexpression cell line, increased, while that of SaOS2FM(H), a high MnSOD overexpression cell line, decreased. Although we assessed PE using a colony-formation assay, time-lapse microscopic observation revealed that cells attached to the flasks had undergone neither apoptosis nor necrosis. Moreover, MnSOD overexpression did not affect cell doubling time. Therefore, MnSOD overexpression might correlate directly with cellular adhesion's effect on PE changes. When L-buthionine-[S,R]-sulfoximine (BSO) was administered to increase the intracellular concentration of hydrogen peroxide, the PEs of both cell lines decreased, and when hydrogen peroxide was eliminated by the administration of sodium pyruvate, only the PE of SaOS2FM(H) increased. The combination of BSO and NO (NOR4 or isosorbide 5-mononitrate) administration synergistically decreased PE in both cell lines. These findings suggest that changes in cellular adhesion properties correlate with the balance between increased hydrogen peroxide levels and decreased superoxide radical levels. This is the first report to indicate that PE and cellular adhesion properties change bidirectionally according to the levels of MnSOD overexpression: first increasing then decreasing as MnSOD activity increases. Our results indicate that PE changes might be decided by the balance between two cytotoxic compounds (decreased superoxide radical levels and increased hydrogen peroxide levels), and that NO loading and increased hydrogen peroxide synergistically reduce PE and cellular adhesion.

Development of a new hybrid gel phantom using carrageenan and gellan gum for visualizing three-dimensional temperature distribution during hyperthermia and radiofrequency ablation.

We developed a new hybrid gel phantom using carrageenan and gellan gum for the purpose of visualizing three-dimensional temperature distribution. The phantom, which contains carrageenan, gellan gum, non-ionic surface active agent, potassium chloride, n-butanol, sodium azide, and water, shows good transparency at room temperature, and has the advantage that the heated region becomes white and opaque due to segregation of the surface active agent. Carrageenan and gellan gum were added to improve the transparency and fragility of the hybrid gel. Potassium chloride was used to adjust the electrical conductivity of the gel to a range of 5-130 MHz, so that it would be equivalent to that of muscle tissue for each frequency used by electromagnetic heating devices. N-butanol was used to adjust the clouding temperature to a range between 45 and 55 degrees C. In the present study we clarified the important properties of the new phantom, and developed formulae for easy determination of the amounts of ingredients necessary for the desired clouding temperature and electric conductivity. The characteristics of this phantom are: a) a solid form to avoid convection by heat conduction; b) sufficient strength without fragility to form a torso without the use of a reinforcing agent; c) high transparency at room temperature and visualization of the heating area as a white turbidity; d) time-lapse and accurate visualization of the changing temperature area without thermal hysteresis; e) electrical properties similar to those of human tissues; f) ease of production; and g) low cost and good safety. This phantom might assist oncologists in their routine checking and study of the performance of electromagnetic heating devices for hyperthermia and radiofrequency ablation.

Diagnostic capabilities of I-131, TI-201, and Tc-99m-MIBI scintigraphy for metastatic differentiated thyroid carcinoma after total thyroidectomy.

We investigated the diagnostic capabilities of I-131, Tl-201, and Tc-99m-MIBI (hexakis-2-methoxyisobutyl- isonitrile) scintigraphy for thyroid cancer metastases after total thyroidectomy over the entire body and for every locus before and after thyroid bed ablation. After total thyroidectomy of thyroid cancer, 36 cases were subjected to I-131 treatment 64 times. They consisted of 17 men and 19 women with 31 papillary carcinomas and 5 follicular carcinomas. Their ages were 22--75(an average of 60.5+/-12.3) years. I-131 scintigraphy(I-131), Tl-201 scintigraphy(Tl-201), and Tc-99m- MIBI scintigraphy (Tc-99m-MIBI) were performed. We defined the metastases as those cases in which serum thyroglobulin (Tg)increased significantly or in which we were able to prove the lesions on CT (computed tomography), MRI (magnetic resonance imaging) or bone scintigram. Three radiology medical specialists visually evaluated each scintigram and calculated the sensitivity, specificity, and likelihood ratio. For whole-body sensitivity, both Tl-201 and Tc-99m-MIBI were high before ablation and I-131 was high after ablation. Before ablation, the negative likelihood ratio was less than 0.1 for Tl-201 and Tc-99m-MIBI, while the positive likelihood ratio was more than 10 for Tl-201. After ablation, the positive likelihood ratio for I-131, Tl-201, and Tc-99m-MIBI was more than 10. The sensitivity of the mediastinum was appropriate, except for I-131 before ablation, and the sensitivity of the lung before and after ablation was inferior for either tracer. The specificity of the cervix for I-131 before ablation was markedly deteriorated, but it increased after ablation.

Well differentiation and intact Smad4 expression are specific features of groove pancreatic ductal adenocarcinomas.

OBJECTIVES: We aimed to select true groove pancreatic ductal adenocarcinomas (GPDACs) and define their specific features. METHODS: We performed histopathologic and immunohistochemical comparisons of 6 GPDACs with 6 duodenal adenocarcinomas (DACs) and 24 conventional pancreatic ductal adenocarcinomas (cPDACs). Both groups were adjusted to ensure similar mean tumor size. RESULTS: Representative loupe image showed prominent duodenal invasion and slight pancreatic invasion. Groove pancreatic ductal adenocarcinomas exhibited different mucins and cytokeratin profiles in DACs, but cPDACs and small branch pancreatic ducts had the same profiles. Histopathologic analysis of GPDACs showed a significantly higher incidence of duodenal invasion and well differentiation than cPDACs, although the incidences of lymph node metastasis, angiolymphatic invasion, and neural invasion were similar. Immunohistochemical analysis of GPDACs showed a significantly lower frequency of abnormal Smad4 immunolabeling, and fewer GPDAC samples exhibited abnormal immunolabeling for MUC1, p16, Smad4, and p53 than cPDACs. CONCLUSIONS: These results suggest that GPDACs arise from small branch pancreatic ducts around accessory pancreatic duct penetrating the groove and duodenum and are distinguishable from DACs. Molecular immunohistochemistry suggests the accumulation of genetic abnormalities during tumor progression is slow in comparison with cPDACs. Thus, the site of PDAC occurrence, such as the border or inner area of the pancreas head, may determine genetic progressivity.

Cepharanthin enhances adriamycin sensitivity by synergistically accelerating apoptosis for adriamycin-resistant osteosarcoma cell lines, SaOS2-AR and SaOS2 F-AR.

Cepharanthin (CEP) is a biscoclaurine alkaloid extracted from Stephania cepharantha Hayata. CEP is reported to inhibit drug resistance by inhibiting P-glycoprotein, a drug efflux pump, and recently to induce apoptosis. In the present study, we examined the effects of CEP as an inhibitor of adriamycin (ADR) resistance on ADR-induced apoptosis and necrosis. First, we established p53-deficient ADR-resistant osteosarcoma cell lines, SaOS2-AR and SaOS2 F-AR. Resistant cells showed a higher level of intracellular glutathione peroxidase activity than parent cells. P-glycoprotein was overexpressed in resistant cells. The intracellular ADR level of resistant cells was lower than that of parent cells. One micro g/ml CEP eliminated the degradation of intracellular ADR of resistant cells; that is, to a level equivalent to that of the parent cells. CEP of 0.5 micro g/ml, which was not cytotoxic when used alone, significantly increased the ADR sensitivity of resistant cells, to a level similar to the parent cell level. Isosorbide 5-mononitrate, a potential nitric oxide-generation agent, combined with CEP further increased the ADR sensitivity of resistant cells, indicating a synergistic effect of CEP and isosorbide 5-mononitrate on ADR cytotoxicity. Time-lapse microscopic observation revealed that ADR dominantly induced apoptosis much more than necrosis for both parent and resistant cells, and that the use of 0.5 micro g/ml CEP with ADR synergistically accelerated apoptosis in resistant cells. Finally, we clarified the property by which CEP synergistically accelerates ADR-induced apoptosis. This property might be a new mechanism that explains how CEP overcomes ADR resistance.

pEYFP-Nuc vector is a useful tool for three-dimensional and time-lapse observation of nuclear morphology of Jurkat cells during apoptosis.

Using a pEYFP-Nuc vector, which contains nucleic acid sequences of a nuclear localization signal, we established a Jurkat-YN cell line that expressed enhanced yellow fluorescent protein (EYFP) in the nucleus. We observed three-dimensional and time-lapse changes in nuclear morphology of Jurkat-YN cells during Fas-induced apoptosis using a confocal laser scanning microscope. The nuclear forms visualized by EYFP were almost equal in quality to those visualized by SYTO59, a nucleic acid stain for living cells. Three-dimensional deformities in the nuclear form were observed during apoptosis before chromatin condensation became apparent, indicating these deformities are characteristic morphological changes of the early stage of apoptosis. In conclusion, the pEYFP-Nuc vector is a useful tool in the time-lapse observation of nuclear morphology of living cells during apoptosis.

Cepharanthine enhances in vitro and in vivo thermosensitivity of a mouse fibrosarcoma, FSa-II, based on increased apoptosis.

Cepharanthine (Ce) is a biscoclaurine alkaloid extracted from Stephania cepharantha Hayata. In our previous study, Ce significantly enhanced thermosensitivity and thereby reduced thermotolerance in vitro, and intra-peritoneal injection of Ce slightly enhanced thermosensitivity in vivo. In the present study, we investigated Ce's effect in vitro on the pattern of cell death after heating and the effect of intra-tumoral injection of Ce on in vivo thermosensitivity using a mouse fibrosarcoma, FSa-II, and C3H/He mice. Ce significantly enhanced the in vitro thermosensitivity of FSa-II cells with heating at 44 degrees C, with increased Ce concentration. Time-lapse microscopic observation of individual cells confirmed that Ce treatment hastened both apoptosis (specifically, apoptotic budding) and necrosis (as indicated by staining with propidium iodide). Staining with annexin V-enhanced green fluorescent protein indicated that Ce used concomitantly with heating significantly increased the proportion of cells in the early stage of apoptosis. Ce combined with heating also significantly increased the proportion of cells with high intracellular caspase-3 activity, as detected by a substrate of caspase-3, PhiPhiLux-G1D2. The intra-tumoral injection of Ce, followed by heating at 44 degrees C, significantly delayed in vivo tumor growth, and this delay increased in a Ce concentration-dependent manner. Ce injected 30 min before heating delayed tumor growth more than Ce injected immediately before heating. These findings suggest the potential of Ce as a thermosensitizer to increase apoptosis of tumor cells.