Reactive oxygen species upregulate expression of muscle atrophy-associated ubiquitin ligase Cbl-b in rat L6 skeletal muscle cells.

Unloading-mediated muscle atrophy is associated with increased reactive oxygen species (ROS) production. We previously demonstrated that elevated ubiquitin ligase casitas B-lineage lymphoma-b (Cbl-b) resulted in the loss of muscle volume (Nakao R, Hirasaka K, Goto J, Ishidoh K, Yamada C, Ohno A, Okumura Y, Nonaka I, Yasutomo K, Baldwin KM, Kominami E, Higashibata A, Nagano K, Tanaka K, Yasui N, Mills EM, Takeda S, Nikawa T. Mol Cell Biol 29: 4798-4811, 2009). However, the pathological role of ROS production associated with unloading-mediated muscle atrophy still remains unknown. Here, we showed that the ROS-mediated signal transduction caused by microgravity or its simulation contributes to Cbl-b expression. In L6 myotubes, the assessment of redox status revealed that oxidized glutathione was increased under microgravity conditions, and simulated microgravity caused a burst of ROS, implicating ROS as a critical upstream mediator linking to downstream atrophic signaling. ROS generation activated the ERK1/2 early-growth response protein (Egr)1/2-Cbl-b signaling pathway, an established contributing pathway to muscle volume loss. Interestingly, antioxidant treatments such as N-acetylcysteine and TEMPOL, but not catalase, blocked the clinorotation-mediated activation of ERK1/2. The increased ROS induced transcriptional activity of Egr1 and/or Egr2 to stimulate Cbl-b expression through the ERK1/2 pathway in L6 myoblasts, since treatment with Egr1/2 siRNA and an ERK1/2 inhibitor significantly suppressed clinorotation-induced Cbl-b and Egr expression, respectively. Promoter and gel mobility shift assays revealed that Cbl-b was upregulated via an Egr consensus oxidative responsive element at -110 to -60 bp of the Cbl-b promoter. Together, this indicates that under microgravity conditions, elevated ROS may be a crucial mechanotransducer in skeletal muscle cells, regulating muscle mass through Cbl-b expression activated by the ERK-Egr signaling pathway.

Granulocyte-colony stimulating factor-producing gallbladder carcinoma-include analysis all case reports: A case report.

INTRODUCTION: It is extremely rare for gallbladder carcinoma to produce granulocyte-colony stimulating factor (G-CSF) and such tumors have a poor prognosis. PRESENTATION OF CASE: A 67-year-old man was admitted with continuous fever. Laboratory tests showed a leukocyte count of 27,980/µL, serum C-reactive protein (CRP) of 9.2mg/dL and serum G-CSF of 225pg/mL. Imaging revealed an irregular gallbladder mass about 90mm in diameter with peripheral enhancement that also involved the liver and transverse colon. G-CSF producing gallbladder carcinoma was diagnosed. We performed cholecystectomy, partial resection of segments 4 and 5 of the liver, partial resection of the transverse colon, and gastrostomy. Histopathological examination showed gallbladder carcinoma (pT3, pN0, M0, G2, and pStage IIIA by the UICC classification, version 7). On immunohistochemical staining, tumor cells were positive for G-CSF. The leukocyte count was normalized postoperatively and fever subsided immediately after surgery. Two months later, the leukocyte count rose to 56,820/µL and metastases to the liver and lymph nodes were detected by CT. Chemotherapy (gemcitabine plus cisplatin) was started and the leukocyte count was normalized after the first course. The patient has continued chemotherapy and has survived for 16 months postoperatively. DISCUSSION: G-CSF producing gallbladder carcinoma has a poor prognosis and most patients die within 12 months of starting therapy. It is rare for patients with recurrence to survive for 16 months after surgery, as in the present case. CONCLUSION: Multidisciplinary therapy (surgery and chemotherapy) may prolong the survival of patients with G-CSF producing gallbladder carcinoma, especially those with recurrence.

[Influence of a radioisotope from out of the effective field of view in a semiconductor single photon emission computed tomography scanner].

Discovery NM 530c (CZT SPECT) is a new single photon emission computed tomography (SPECT) scanner using a cadmium-zinc-telluride (CZT) solid-state semiconductor detector technology. Due to multi-pinhole collimator design of this system, each CZT detector facing different direction and be able to get incidence radioactivity from radioisotopes (RIs) existing outside of effective field of view (EFOV). The purpose of this study is to verify its impact and compare it to a conventional Anger-type SPECT scanner (Discovery NM/CT 670 pro). We used (99m)TcO4(-) as radiation source and set it outside of the EFOV at several different positions (height and angle) and scanned by both the cameras. As a result, CZT SPECT got more influence compared to Anger-type SPECT. The impact was different according to its height. When using other RIs in CZT SPECT room, it is important to confirm the appropriate position against CZT SPECT during scan.

[A case of advanced gastric cancer with tumor embolus in the portal vein and liver metastasis responding to S-1 plus cisplatin chemotherapy].

A 67-year-old woman presented with anemia. Computed tomography and upper gastrointestinal endoscopy revealed a primary gastric cancer with tumor embolus in the portal vein, liver metastasis, lymph node metastasis, and pancreatic involvement. Because curative surgery was deemed impossible, we started chemotherapy using S-1 (120 mg/m(2)/day for 3 weeks, followed by discontinuation for 2 weeks) plus cisplatin (80 mg/m(2)/day on days 1 and 8). After 4 courses of chemotherapy, the tumor embolus in the portal vein, liver metastasis, lymph node metastasis, and pancreatic involvement had resolved. Therefore, we performed distal gastrectomy. Histological examination revealed ypT1a, ly0, v0, ypN0 (0/49), ypCY0, ypStage IA, with a two-grade histological change in the main tumor after chemotherapy. Postoperatively, she underwent adjuvant chemotherapy with S-1 for 1 year (120 mg/m(2)/day for 4 weeks, followed by discontinuation for 2 weeks). At the 30-month follow-up after the adjuvant chemotherapy, she had no recurrence.

Histopathologic profiles of coronary atherosclerosis by myocardial bridge underlying myocardial infarction.

OBJECTIVE: Anatomic properties of myocardial bridge (MB) are sometimes responsible for myocardial infarction (MI) through the changes in the atherosclerosis distribution in the left ascending coronary artery (LAD). The purpose of this study was to investigate histopathologic profiles of atherosclerotic lesions resulting from the MB presence in the LAD in the MI cases. METHODS: In 150 consecutive autopsied MI hearts either with MBs [MI(+)MB(+); n = 67] or without MBs [MI(+)MB(-); n = 83] and 100 normal hearts with MBs [MI(-)MB(+)], LADs were consecutively cross-sectioned at 5-mm intervals. The most advanced intimal lesion and unstable plaque-related lesion characteristics (UPLCs) in each section were histopathologically evaluated in conjunction with the anatomic properties of the MB, such as its thickness, length, location, and MB muscle volume burden (MMV: the total volume of MB thickness multiplied by MB length). RESULTS: The MB showed a significantly greater thickness (P = 0.0090), length (P = 0.0300), and MMV (P = 0.0019) in MI(+)MB(+) than in MI(-)MB(+). Mean age of acute MI cases was significantly younger (P = 0.0227) in MI(+)MB(+) than in MI(+)MB(-). Frequency of plaque fissure/rupture in the proximal LAD was significantly higher in acute MI cases of MI(+)MB(+) than in MI(+)MB(-). UPLCs tended to be located proximally in MI(+)MB(+) and frequent 2.0 cm or more proximal to the MB entrance in MI(+)MB(+). CONCLUSION: In MI(+)MB(+), UPLCs tend to be located more proximally, and a plaque in the LAD proximal to the MB is prone to rupture, resulting in MI at younger age.

Novel method for quantitative evaluation of cardiac amyloidosis using (201)TlCl and (99m)Tc-PYP SPECT.

OBJECTIVE: The degree of myocardial technetium-(99m)-pyrophosphate ((99m)Tc-PYP) accumulation in cardiac amyloidosis is conventionally evaluated by the PYP score. This method involves qualitative visual evaluation on two-dimensional images. Here, we performed three-dimensional quantitative analysis using software developed in our laboratory. METHODS: We performed dual myocardial imaging using thallium-(201)-chloride ((201)Tl-Cl) and (99m)Tc-PYP in cases of suspected cardiac amyloidosis and calculated the PYP accumulation rates of all myocardial pixels showing (99m)Tc-PYP accumulation. We defined this procedure as quantitative evaluation of the degree of (99m)Tc-PYP accumulation in the myocardium. Patients were divided into two groups with and without a diagnosis of cardiac amyloidosis, and we examined the PYP accumulation rates in both groups. In addition, we examined the PYP scores of the two groups by conventional qualitative evaluation. RESULTS: The PYP scores of the cardiac amyloidosis group were significantly higher than those of the other group. The PYP accumulation rates of the cardiac amyloidosis group were significantly higher than those of the other group. There were significant differences in the PYP accumulation rate and PYP score between the two groups. There was considered to be a threshold between the two groups in the case of the PYP accumulation rate. CONCLUSIONS: When the threshold of the PYP score was defined as 3+ and that of the PYP accumulation rate as 41.5 %, the sensitivity of the PYP score and PYP accumulation rate was 84.6 %. However, the specificity of the PYP accumulation rate was higher than that of the PYP score. Quantitative evaluation by the PYP accumulation rate of the degree of (99m)Tc-PYP accumulation in the myocardium may be useful in the diagnosis of cardiac amyloidosis.

Atherosclerosis prevention in youth.

Atherosclerosis-associated circulatory disturbance is one of the most important global issues. In patients with atherosclerosis, eccentric intimal thickening and lipid deposition progress over a long period (at least 20 to 30 years). On the other hand, in patients with atherosclerosis-associated circulatory disturbance represented by myocardial infarction, the direct cause of death is thrombus formation rather than marked stenosis; wall destruction may lead to a fatal outcome. In the future, atherosclerosis susceptibility, that is, intrinsic genes, should be investigated.

[Evaluation of shortened protocol of graph plot method with ¹²³I-IMP].

The (123)I-IMP Graph Plot is a convenient method of analyzing cerebral blood flow without blood sampling. Data acquisition requires 45 minutes after the infusion of (123)I-IMP because the method is matched to the protocol of autoradiography (conventional method). However, we think that those protocols do not have to be matched because those theories are different. Therefore, we contrived a protocol for shortening that time by beginning SPECT data acquisition earlier and shortening the acquisition time compared to the conventional method. We compared count ratios of the decreased area to an area of the healthy side, quantitative values, and results of statistical analysis of the shortened protocol and the conventional method for cases of cerebral infarction and of dementia with Lewy bodies (DLB). Some count ratios of the shortened protocol were inferior to those of the conventional method, but the degrees did not affect the clinical diagnosis. In the other areas and cases also, the differences did not affect the clinical diagnosis. In addition to the results of this study, some previous reports have described that early SPECT images after infusion show the true cerebral blood flow. Therefore, we judged that this shortened protocol can be used as a clinical protocol.

Angiosarcoma-like metastatic carcinoma of the liver.

Two cases of metastatic carcinoma strikingly simulating angiosarcoma in the liver are described. The first case was a 53-year-old female with cystic liver tumors which were found 22 months after surgery for ureteral cancer. The second case was an 81-year-old female with multiple tumors in the liver and the pancreas, and a post-mortem examination was carried out. She had undergone surgery for skin cancer three years before. Both cases had an angiosarcoma-like appearance macroscopically and microscopically. Immunohistochemically, the tumor cells of both cases were negative for CD31, CD34, and Factor VIII-related antigen and positive for several types of cytokeratin, suggesting that they were not angiosarcomas but carcinomas. Angiosarcoma is the most common sarcoma arising in the liver. Thus, metastatic carcinoma, which resembles angiosarcoma, might be mistaken for angiosarcoma.

Anatomic properties of myocardial bridge predisposing to myocardial infarction.

BACKGROUND: A myocardial bridge (MB) that partially covers the course of the left anterior descending coronary artery (LAD) sometimes causes myocardial ischemia, primarily because of hemodynamic deterioration, but without atherosclerosis. However, the mechanism of occurrence of myocardial infarction (MI) as a result of an MB in patients with spontaneously developing atherosclerosis is unclear. METHODS AND RESULTS: One hundred consecutive autopsied MI hearts either with MBs [MI(+)MB(+) group; n=46] or without MBs (n=54) were obtained, as were 200 normal hearts, 100 with MBs [MI(-)MB(+) group] and 100 without MBs. By microscopy on LADs that were consecutively cross-sectioned at 5-mm intervals, the extent and distribution of LAD atherosclerosis were investigated histomorphometrically in conjunction with the anatomic properties of the MB, such as its thickness, length, and location and the MB muscle index (MB thickness multiplied by MB length), according to MI and MB status. In the MI(+)MB(+) group, the MB showed a significantly greater thickness and greater MB muscle index (P<0.05) than in the MI(-)MB(+) group. The intima-media ratio (intimal area/medial area) within 1.0 cm of the left coronary ostium was also greater (P<0.05) in the MI(+)MB(+) group than in the other groups. In addition, in the MI(+)MB(+) group, the location of the segment that exhibited the greatest intima-media ratio in the LAD proximal to the MB correlated significantly (P<0.001) with the location of the MB entrance, and furthermore, atherosclerosis progression in the LAD proximal to the MB was largest at 2.0 cm from the MB entrance. CONCLUSIONS: In the proximal LAD with an MB, MB muscle index is associated with a shift of coronary disease more proximally, an effect that may increase the risk of MI.

Application of liquid-based preparation to fine needle aspiration cytology in breast cancer.

OBJECTIVE: To examine the performance of liquid-based cytology (LBC) in breast cytology to confirm the diagnosis of carcinoma. STUDY DESIGN: Using cell clusters directly scratched from surgically removed tumor masses, we examined the immunocytochemistry, molecular biology and cytomorphology of the specimens. RESULTS: LBC was very useful for gene analysis and evaluating the immunocytochemistry. The cytologic features of LBC were slightly different from those ofa conventional aspiration cytology smear. CONCLUSION: LBC is a promising method for improving the standardization ofpreparations in breast cytology, although care should be taken to account for its characteristic cytologic features. The quantitative analysis of HER-2 mRNA correlated with the results of immunohistochemistry.

[Examination of axial zygomatic arch radiographs using the bisector method].

An axial radiograph of the zygomatic arch is taken in cases of patients with facial traumatic injury. Maintaining the patient's head in the retroflex position to take such axial radiographs is sometimes difficult because of medical conditions. In addition, since different positioning techniques for retroflexion are used by radiological technologists, the visibility of the zygomatic arch was poorly in reproduced. We contrived a novel technique for use in taking a zygomatic arch radiograph. We call it the "bisector method," and it does not require the retroflex position. We can take a zygomatic arch radiograph equal in quality to conventional axial radiographs (retroflex position) by exposing X-rays perpendicularly to the bisector of the angle between the casette and the zygomatic arch. This bisector method is relatively easy in that it does not require either the retroflex position or the expertise of a radiological technologist.

[Imaging properties of SAR-reduced sequences in 3T-MR system].

Reduction of the specific absorption ratio (SAR) is essential in MR systems with high magnetic fields. Therefore, a variety of SAR-reduced sequences are available on modern 3T-MR systems, and some sequences can be applied under the same conditions as conventional sequences. The purpose of this study was to compare the imaging properties of a phantom acquired by SAR-reduced sequences with those of conventional sequences. Our data showed that effective relaxation time differed in each of the sequences, because the echo space and RF pulse of the SAR-reduced sequences varied from those of the conventional sequences. Further, the image contrast of the SAR-reduced sequences differed from the image contrast of the conventional ones.

[Examination of retention index in brain tumor 201Tl scintigraphy using iNRT].

The retention index (R.I.) in Thalliun-201-chloride (201Tl-Cl) brain tumor single photon emission computed tomography (SPECT) imaging analysis is useful in assessing grades of malignancy. When the T/N ratio that influences the R.I. is calculated, it is ideal to draw an ROI on the same slices and coordinates of both early and delayed transaxial images. However, in many institutions, the image slice and ROI are determined manually by operators (i.e., by the conventional method). The Co-registration function within iNRT software enables an ROI to be automatically drawn. In this study, we obtained the R.I. from six brain tumor patients employing two different methods (i.e., Co-registration and the conventional method). Each R.I. was obtained from the same personal computer that was used to transfer the transaxial images. We then compared and evaluated the six R.I. values of seven operators. (2 radiologists and 5 radiological technologists). The results showed that the R.I. values did not differ significantly between the two methods, except in two cases. Furthermore, the coefficient of variation (CV) of the R.I. obtained from the Co-registration method was lower than the conventional method in all cases. The variations among operators were also smaller in the Co-registration method. These outcomes may have been due to the fact that in the conventional method it is difficult to identify the same slices of early and delayed images. Therefore, there were differences among operators in their ways of specifying positions of the ROI. This may have resulted in differences in the number of counts/pixel of the ROI. In conclusion, the R.I. obtained by using the Co-registration method would be more useful for improving diagnostic accuracy in (201)Tl-Cl brain tumor SPECT imaging than the conventional method.

[Examination of normal database in three-dimensional SSP].

The results of statistical image analyses, such as three-dimensional stereotactic surface projection (3D-SSP), are affected by the normal database (NDB) used. As a general rule, each institution must establish its own NDB for analysis. However, the numbers of institutions with the ability to establish their own NDB are limited. Therefore, it would be ideal if an NDB suitable for other institutions could be established from standard volunteer data gathered at a single institution. In this study, we examined the validity of establishing an NDB suitable for other institutions from the standard volunteer data gathered at our institution, using E.CAM (used as a reference) and IRIX. We established three NDBs. One was established from E.CAM, and the other two were from IRIX (in which overall FWHM was the same as that of E.CAM with Butterworth filter cutoff frequency and using an identical algorithm for image-reconstruction of E.CAM). We then compared the mean values and standard deviations of the NDBs, and also examined the effects on the Z-score map. We determined that the NDB that had been established was suitable for other institutions, using identical FWHM (resolution of SPECT imaging) and an algorithm for image-reconstruction.

Interobserver variation in the histopathological assessment of nonalcoholic steatohepatitis.

Pathological assessment is considered as the gold standard for the diagnosis of nonalcoholic steatohepatitis (NASH). However, there is no agreement of histological features required for the diagnosis of NASH. In the present study, eight experienced hepatopathologists read liver biopsy specimen slides of 21 cases of NASH and suspected NASH independently and were asked to assess histopathological features and render a diagnosis. Interobserver variation among pathologists was evaluated by kappa statistics. Significant, good agreement was present in evaluation of the extent of steatosis and grade of fibrosis. Agreement was moderate concerning the localization of steatosis, localization of fibrosis, and glycogen nuclei. Only slight or poor agreement was seen in evaluation of type of steatosis, ballooning, intralobular necroinflammatory change, portal inflammation, and degree of neutrophilic infiltration. Thus the agreement varied for histological variables. Significant, moderate agreement was seen in the diagnosis of NASH but agreement was poor in the diagnosis of suggestive NASH. The agreement for the diagnosis of NASH was not high as for the individual histological findings that were thought to be the basis of the diagnosis. In conclusion, some histological features in NASH might prove useful for the development of a standardized and reliable pathological diagnosis and scoring system.