A natural infection by the red sea bream iridovirus-type Megalocytivirus in the golden mandarin fish Siniperca scherzeri.

An outbreak of a Megalocytivirus infection was found in the golden mandarin fish Siniperca scherzeri during September and October 2016, in Korea. Phylogeny and genetic diversity based on the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes showed a new strain. Designated as GMIV, this strain derived from the golden mandarin fish was suggested to belong to the red sea bream iridovirus (RSIV)-subgroup I. Additionally, this train clustered with the ehime-1 strain from red sea bream Pagrus major in Japan and was distinguished from circulating isolates (RSIV-type subgroup II and turbot reddish body iridovirus [TRBIV] type) in Korea. The infection level, evaluated by qPCR, ranged from 8.18 × 102 to 7.95 × 106  copies/mg of tissue individually, suggesting that the infected fish were in the disease-transmitting stage. The diseased fish showed degenerative changes associated with cytomegaly in the spleen as general sign of Megalocytivirus infection. The results confirm that the RSIV-type Megalocytivirus might have crossed the environmental and species barriers to cause widespread infection in freshwater fish.

Comparison of mode of failure between primary and revision total knee arthroplasties.

INTRODUCTION: Cognizance of common reasons for failure in primary and revision TKA, together with their time course, facilitates prevention. However, there have been few reports specifically comparing modes of failure for primary vs. revision TKA using a single prosthesis. The goal of the study was to compare the survival rates, modes of failure, and time periods associated with each mode of failure, of primary vs. revision TKA. HYPOTHESIS: The survival rates, modes of failure, time period for each mode of failure, and risk factors would differ between primary and revision TKA. MATERIAL AND METHODS: Data from a consecutive cohort comprising 1606 knees (1174 patients) of primary TKA patients, and 258 knees (224 patients) of revision TKA patients, in all of whom surgery involved a P.F.C® prosthesis (Depuy, Johnson & Johnson, Warsaw, IN), was retrospectively reviewed. The mean follow-up periods of primary and revision TKAs were 9.2 and 9.8 years, respectively. RESULTS: The average 10- and 15-year survival rates for primary TKA were 96.7% (CI 95%,±0.7%) and 85.4% (CI 95%,±2.0%), and for revision TKA 91.4% (CI 95%,±2.5%) and 80.5% (CI 95%,±4.5%). Common modes of failure included polyethylene wear, loosening, and infection. The most common mode of failure was polyethylene wear in primary TKA, and infection in revision TKA. The mean periods (i.e., latencies) of polyethylene wear and loosening did not differ between primary and revision TKAs, but the mean period of infection was significantly longer for revision TKA (1.2 vs. 4.8 years, P=0.003). DISCUSSION: Survival rates decreased with time, particularly more than 10 years post-surgery, for both primary and revision TKAs. Continuous efforts are required to prevent and detect the various modes of failure during long-term follow-up. Greater attention is necessary to detect late infection-induced failure following revision TKA. LEVEL OF EVIDENCE: Case-control study, Level III.

Nonthermal plasma induces head and neck cancer cell death: the potential involvement of mitogen-activated protein kinase-dependent mitochondrial reactive oxygen species.

Nonthermal plasma (NTP) is generated by ionization of neutral gas molecules, which results in a mixture of energy particles including electrons and ions. Recent progress in the understanding of NTP has led to its application in the treatment of various diseases, including cancer. However, the molecular mechanisms of NTP-induced cell death are unclear. The purpose of this study was to evaluate the molecular mechanism of NTP in the induction of apoptosis of head and neck cancer (HNC) cells. The effects of NTP on apoptosis were investigated using MTT, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling, Annexin V assays, and western blot analysis. The cells were examined for production of reactive oxygen species (ROS) using DCFCA or MitoSOX staining, intracellular signaling, and an animal model. NTP reduced HNC cell viability in a dose-dependent manner and induced apoptosis. NTP resulted in alteration of mitochondrial membrane potential and accumulation of intracellular ROS generated from the mitochondria in HNC cells. Blockade of ROS production by N-acetyl-L-cysteine inhibited NTP-induced apoptosis. NTP led to the phosphorylation of c-JUN N-terminal kinase (JNK) and p38, but not extracellular-regulated kinase. Treatment with JNK and p38 inhibitors alleviated NTP-induced apoptosis via ROS generation. Taken together, these results show that NTP induced apoptosis of HNC cells by a mechanism involving MAPK-dependent mitochondrial ROS. NTP inhibited the growth of pre-established FaDu tumors in a nude mouse xenograft model and resulted in accumulation of intracellular ROS. In conclusion, NTP induced apoptosis in HNC cells through a novel mechanism involving MAPK-mediated mitochondrial ROS. These findings show the therapeutic potential of NTP in HNC.

Efficacy and safety using autotransfusion system with postoperative shed blood following total knee arthroplasty in haemophilia.

The purpose of this study was to evaluate the efficacy and safety of postoperative wound drain salvage and autotransfusion system in haemophilic patients undergoing elective total knee arthroplasty (TKA). No literature exists on reinfusing drained blood in patient with haemophilia undergoing TKA. Eighty-eight knees of 66 patients received cemented TKA due to end-stage haemophilic arthropathy (group I; with autotransfusion in 59 knees, group II; without autotransfusion in 29 knees). In group I, the postoperative shed blood was transfused within 6 h after surgery. The amount of blood drainage and reinfused blood, rate and amount of allogenic transfusion, postoperative change of haemoglobin level, prothrombin time (PT) and activated partial thromboplastin time were analysed. The mean postoperative blood drainage was 932 ± 479 mL in group I and 830 ± 492 mL in group II (P > 0.05). The mean volume of blood reinfused was 530 ± 265 mL in group I. Allogenic transfusion was needed in six knees (10.2%) of group I and eight knees (27.6%) of group II (P = 0.036). The mean volume of allogenic transfusion was 480 ± 49 mL in group I and 1041 ± 691 mL in group II (P > 0.05). Changes of all the laboratory results before and after TKA showed no statistically significant difference except PT was prolonged in group I (P = 0.008) at postoperative day 1. Moreover, there was no significant complication related to either reinfusion or allogenic transfusion in both groups. This study showed that reinfusion of drained blood is a simple, safe and efficacious method in patients with haemophilia undergoing TKA.

Computer-navigated total knee arthroplasty in haemophilic arthropathy.

Total knee arthroplasty (TKA) in end-stage haemophilic arthropathy is complex and challenging due to the altered bony anatomy, arthrofibrosis and muscle contractures. Computer navigation is especially advocated in patients with deformity or altered anatomy to improve alignment and to assist in ligament balancing. The objective of this study was to evaluate the results of computer-navigated TKA in haemophilic arthropathy. A consecutive series of computer-assisted TKA for the end-stage haemophilic arthropathy between February 2007 and December 2009 were evaluated. A total of 27 TKA were performed in 25 patients. Pre- and postoperative full-length weight-bearing radiographs were assessed for the axial limb alignment. The orientation of the components was measured on anteroposterior radiographs. Clinically, Knee Society score and Short Form-36 were evaluated. The mechanical axis of the leg was within a range of ±3° varus/valgus in 92% of the TKA. The coronal alignment of the femoral and tibial components was within a range of ±3 degrees in 96% of the knees. The clinical outcomes were significantly improved after the operation. There were no complications specific to the computer navigation. Computer-navigated TKA helps in restoring the mechanical axis and improves accuracy of orientation of the components in patients with end-stage haemophilic arthropathy. Potential benefits in long-term outcome require further investigation.

Comparison of chest digital tomosynthesis and chest radiography for detection of asbestos-related pleuropulmonary disease.

AIM: To compare chest digital tomosynthesis (DTS) with digital radiography for the detection of asbestos-related pleuropulmonary disease. MATERIALS AND METHODS: The institutional review board approved this study and all participants gave informed consent. Forty-five participants with a history of asbestos exposure were examined with DTS and radiography. Low-dose multidetector computed tomography (CT) in the prone position served as the reference. Two observers evaluated all images for the presence of pleural abnormalities and asbestosis. Interobserver agreement was analysed by using the k statistic. Diagnostic performance of the two imaging methods was compared using McNemar's test. RESULTS: Interobserver agreement regarding DTS findings was moderate to very good (k = 0.544-0.846) and superior to the radiographic findings (k = 0.236-1.000). Overall, the diagnostic accuracy of DTS for the lesion detection was significantly better than with radiography (all p < 0.05, except that for the comparison of diagnostic accuracy of DTS versus radiographic detection of left diaphragmatic plaques and asbestosis). DTS was more sensitive than radiography for the detection of asbestosis (82% versus 27%, p = 0.031). CONCLUSION: DTS is more accurate than radiography in the detection of pleural plaques and more sensitive than radiography in the detection of asbestosis. Interobserver agreements with respect to the DTS findings were superior to the radiographic findings.

Influence of long-term statin use in type 2 diabetic patients on thyroid nodularity in iodine-sufficient area.

Statins have marked beneficial effects on lipid profile, but also have pleiotropic actions. A previous study in an iodine-deficient area suggested that statin use is associated with reduced thyroid volume and nodularity. We performed this study to investigate how long-term statin use in type 2 diabetic patients affects thyroid nodularity in iodine-sufficient area.We recruited euthyroid type 2 diabetic patients, receiving statin therapy continuously for at least 5 years (statin group) and, age and sex matched statin-naive type 2 diabetic patients (control group). Subjects with past history of cancer, thyroid disease or treatment with lithium or amiodarone; family history of thyroid cancer; palpable goiter or thyroid nodule, and/or positive thyroperoxidase antibody were excluded. The prevalence, number, and volume of thyroid nodules, size of thyroid were evaluated in all subjects by high resolution ultrasound.Prevalence of non-palpable thyroid nodules of statin group (n=70) and control group (n=98) were 51 and 53%, respectively. There was no difference of prevalence, number, and volume of non-palpable thyroid nodules and size of thyroid between statin and control group. But, the patients aged between 60 and 65 years from statin group showed lower prevalence of non-palpable thyroid nodules than the patients with same age interval from control group (4 out of 12 patients, 33%, statin group; 19 out of 27 patients, 70%, control group; P=0.04).Long-term statin use in elderly type 2 diabetic patients was associated with lesser prevalence of thyroid nodules in an iodine-sufficient area. Our data might support a possible antiproliferative effect of statins on thyroid in old type 2 diabetic patients. But, the effect was not as strong as that in an iodine-deficient area and further studies with enough numbers of subjects and revised design will be needed.

Prognostic significance of molecular subtype in T1N0M0 breast cancer: Korean experience.

PURPOSE: Gene expression profiling studies have identified several breast cancer subtypes associated with markedly different clinical outcomes. In general, patients with stage I breast cancer have excellent outcomes. We assessed the clinicopathological characteristics and outcomes of patients with T1N0M0 breast cancer according to molecular subtype. METHODS: Seven hundred and sixty-two T1N0M0 breast cancer patients undergoing curative surgery between January 1990 and December 2007 were analyzed. Subtypes were classified according to hormone receptor (HR) and human epidermal growth factor receptor-2 (HER2) status as follows: HR+/HER2-, HR+/HER2+, HR-/HER2- (triple-negative, TN), and HR-/HER2+. RESULTS: The distribution of subtypes was HR+/HER2-, 56.6%; HR+/HER2+, 10.1%; TN, 20.1%; and HR-/HER2+, 13.3%. Marked differences were observed among subtypes in multifocality/multicentricity, histological grade, extensive intraductal components, p53 expression and the Ki-67 index. There were differences in recurrence-free survival and overall survival among patients with different molecular subtypes (log-rank p < 0.001 and 0.024, respectively). By multivariate analysis, lymphovascular invasion and classification of molecular subtype were independent predictors of recurrence (p = 0.003 and 0.043, respectively). The TN subtype showed significantly worse recurrence-free survival compared to the HR+/HER2- subtype (hazard ratio, 4.54; 95% confidence interval, 1.60-12.86; p = 0.004). CONCLUSION: Patients with T1N0M0 breast cancer, a group with generally favorable clinical outcomes, had prognoses that were associated with the molecular subtype. The TN subtype was an independent predictor for recurrence in patients with T1N0M0 breast cancer.

Peri-acetabular rotational osteotomy with concomitant hip arthroscopy for treatment of hip dysplasia.

Reconstructive acetabular osteotomy is a well established and effective procedure in the treatment of acetabular dysplasia. However, the dysplasia is frequently accompanied by intra-articular pathology such as labral tears. We intended to determine whether a concomitant hip arthroscopy with peri-acetabular rotational osteotomy could identify and treat intra-articular pathology associated with dysplasia and thereby produce a favourable outcome. We prospectively evaluated 43 consecutive hips treated by combined arthroscopy and acetabular osteotomy. Intra-operative arthroscopic examination revealed labral lesions in 38 hips. At a mean follow-up of 74 months (60 to 97) the mean Harris hip score improved from 72.4 to 94.0 (p < 0.001), as did all the radiological parameters (p < 0.001). Complications included penetration of the joint by the osteotome in one patient, a fracture of the posterior column in another and deep-vein thrombosis in one further patient. This combined surgical treatment gave good results in the medium term. We suggest that arthroscopy of the hip can be performed in conjunction with peri-acetabular osteotomy to provide good results in patients with symptomatic dysplasia of the hip, and the arthroscopic treatment of intra-articular pathology may alter the progression of osteoarthritis.

Radioisotope synoviorthesis with Holmium-166-chitosan complex in haemophilic arthropathy.

Radiosynoviorthesis is a safe and easy method for synovectomy in haemophilic arthropathy. Various agents have been used in radiosynoviorthesis, especially newly developed agent Holmium-166-chitosan complex has good clinical outcome. This study analysed clinical results and radiologic evaluation of radioisotope synoviorthesis using Holmium-166-chitosan complex in haemophilic arthropathy. From March 2001 to December 2003, 58 radiosynoviorthesis were performed in 53 haemophiliacs. The average age at procedure was 13.8 years. The Arnold and Hilgartner stage of the patients was from I to IV. Holmium-166-chitosan complex was injected in 31 ankle joints, 19 elbow joints and 8 knee joints. Average follow-up was 33 months since primary procedure. The range of motion of each joint, frequency of intra-articular bleeding and factor dose used were analysed for clinical assessment. There was no significant improvement of range of motion in affected joints. After procedure, the average frequency of bleeding of the elbow joint has decreased from 3.76 to 0.47 times per month, the knee joint from 5.87 to 1.12 times per month, and the ankle joint from 3.62 to 0.73 times per month respectively (P < 0.05). After treatment, the average coagulation factor dose injected was significantly decreased to 779.3 units per month from 2814.8 units per month before treatment (P < 0.001). Radioisotope synoviorthesis with Holmium-166-chitosan complex in haemophilic arthropathy is a very safe and simple procedure with the expectation of a satisfactory outcome without serious complication. It has excellent bleeding control effect on target joint and the need for substitution of coagulation factor concentrate can be reduced.

Hyperfractionated radiotherapy with concurrent chemotherapy for para-aortic lymph node recurrence in carcinoma of the cervix.

PURPOSE: To evaluate efficacy, toxicity, and patterns of relapse in patients treated with hyperfractionated radiotherapy (HFRT) with concurrent chemotherapy for para-aortic lymph node (PALN) recurrence of cervical carcinoma. METHODS AND MATERIALS: Between September 1997 and October 2000, 12 cervical carcinoma patients with isolated PALN recurrence who had previously received radical or postoperative radiotherapy were treated with HFRT and concurrent chemotherapy. The initial FIGO stage was Stage IB in 4 (33%) patients, Stage IIA in 2 (17%), and Stage IIB in 6 (50%). The radiation field encompassed the gross recurrent PALN with the superior margin at the upper end of the T12 body and the inferior margin between L5 and S1. The fractionated dose was 1.2 Gy in 2 daily fractions, and the median total dose was 60 Gy. The weekly concurrent chemotherapy consisted of paclitaxel in 11 patients and cisplatin in 1. The median number of cycles of chemotherapy was 5. RESULTS: The latent period to PALN recurrence from the time of initial treatment for all patients ranged from 2 to 92 months (median: 12 months). One month after treatment, the clinical tumor response evaluated was complete in 33% (4/12) and partial in 67% (8/12). The 3-year overall survival rate and median survival were 19% and 21 months, respectively. The latent period to PALN recurrence was the only significant prognostic factor; the median survival of patients who relapsed in < or =24 months from the initial treatment of cervical carcinoma was 13 months vs. 45 months for those relapsed at >24 months (p = 0.026). Grade 3-4 hematologic toxicity developed in 2 patients. Six (50%) patients experienced Grade 2 nausea. There were no late gastrointestinal or neurologic complications during the follow-up period. Subsequent distant metastases after PALN treatment developed in 58% (7/12). CONCLUSION: HFRT of 60 Gy to PALN with concurrent chemotherapy could be regarded as an effective treatment modality without significant acute or late toxicity. Patients with a latent period >24 months until PALN recurrence had a more favorable survival rate than those with a latent period

Imaging of occupational lung disease.

Occupational lung disease comprises a wide variety of disorders caused by the inhalation or ingestion of dust particles or noxious chemicals. These disorders include pneumoconiosis, asbestos-related pleural and parenchymal disease, chemical pneumonitis, occupational infection, hypersensitivity pneumonitis, and organic dust toxic syndrome. Most of these disorders produce diffuse lung disease. Although many of the disorders can be detected at chest radiography, high-resolution computed tomography (CT) has been shown to be superior to chest radiography in depicting parenchymal, airway, and pleural abnormalities. Some occupational lung diseases have characteristic radiologic features suggesting the correct diagnosis, whereas in others, a combination of clinical features, related occupational history, radiologic findings, and literature supporting an association between the exposure and the disease process is required for diagnosis. With advances in chest radiology, including high-resolution CT, radiologists play a key role in the clinical evaluation of occupational lung diseases and should continue their involvement in the diagnosis and treatment of these diseases.

Atrial fibrillation surgery simplified with cryoablation to improve left atrial function.

BACKGROUND: The Maze procedure restores atrial fibrillation to normal sinus rhythm. However, concurrent left atrial functional recovery is not always achieved. To address this limitation, a modification using linear cryoablation is described. METHODS: Between July 1997 and December 1999, 83 patients received atrial fibrillation surgery in association with mitral valve surgery with or without additional concurrent procedures by either the conventional technique, group I (n = 30) or the modified technique, group II (n = 53). Onset of sinus conversion and echocardiographic assessment of postoperative left ventricular function, left atrial size, and mitral A-wave velocity were compared in the early postoperative period and 6 months after surgery. RESULTS: Sinus conversion occurred significantly earlier in group II, 2.4 +/- 5 days versus group I, 7.0 +/- 10 days. The mean transmitral A-wave velocity and the incidence of A-wave appearance in the early postoperative period and 6 months postoperatively were greater in group II than group I. CONCLUSIONS: With the current modification, restoration of sinus rhythm and superior left atrial contractile function occurred earlier than with the standard Maze III technique.

How reliable is the ultrasound densitometer for community screening to diagnose osteoporosis in spine, femur, and forearm?

Dual X-ray absorptiometry (DXA) is the most widely used noninvasive, quantitative diagnostic method of measuring bone density, but it exhibits several limitations for the purpose of general screening and community-based study. Thus, we evaluated the feasibility of the ultrasound densitometer SoundScan Compact for screening the osteoporosis community. We compared the reliability of the SoundScan Compact's measurement to predict both osteopenia and osteoporosis at the spine, femur, and forearm measured by the XR-36. World Health Organisation criteria and Korean reference were used for the diagnosis of osteopenia and osteoporosis. Of the 1827 subjects screened in the community, we randomly selected and invited 106 subjects to our university hospital center for clinical epidemiology for further evaluation of their bone health using DXA. The site-specific relationships among speed of sound and bone mineral density at L2-L4 spine, femoral neck, trochanter, Ward's triangle, distal radius + ulna, proximal radius + ulna, and proximal radius were highly correlated with r values of 0.633, 0.632, 0.674, 0.632, 0.668, 0.78, and 0.777 respectively (all p < 0.0001). Of the 42 subjects defined as normal in the spine by SoundScan Compact's, 27 (64.3%) were also in agreement as the normal by the XR-36. However, an agreement of 43.5% (10/23) in osteopenia and 53.7% (22/41) in osteoporosis was less sensitive. Reliability tested by kappa statistics (kappa = 0.35; t = 5.3; p < 0.0001) revealed that the degree of agreement was not coincidental. On the femur, 54.8% (23/42) agreed as normal, 73.9% (17/23) as osteopenia, and 39% (16/41) as osteoporosis (kappa = 0.33; t = 5.5; p < 0.0001). Higher reliability was found in the forearm, with a kappa value of 0.48, t = 7.2, and p < 0.0001. A magnitude of agreement was also greater in the forearm, with 83.3% (35/42) agreement in normal, 52.2% (12/23) in osteopenia, and 64% (16/25) in osteoporosis. Although the SoundScan Compact's measures different bone properties and measurements are taken at different sites, it is a very useful instrument with a high correlation and high degree of variation and reliability to predict bone health at other sites such as the spine, femur, and forearm. The ultrasound densitometer such as SoundScan Compact's merits its use for community screening and in the primary care setting to detect osteopenia and osteoporosis for the purpose of early detection and prevention.

IKKalpha controls formation of the epidermis independently of NF-kappaB.

The IKKalpha and IKKbeta catalytic subunits of IkappaB kinase (IKK) share 51% amino-acid identity and similar biochemical activities: they both phosphorylate IkappaB proteins at serines that trigger their degradation. IKKalpha and IKKbeta differ, however, in their physiological functions. IKKbeta and the IKKgamma/NEMO regulatory subunit are required for activating NF-kappaB by pro-inflammatory stimuli and preventing apoptosis induced by tumour necrosis factor-alpha (refs 5,6,7,8,9,10,11). IKKalpha is dispensable for these functions, but is essential for developing the epidermis and its derivatives. The mammalian epidermis is composed of the basal, spinous, granular and cornified layers. Only basal keratinocytes can proliferate and give rise to differentiated derivatives, which on full maturation undergo enucleation to generate the cornified layer. Curiously, keratinocyte-specific inhibition of NF-kappaB, as in Ikkalpha-/- mice, results in epidermal thickening but does not block terminal differentiation. It has been proposed that the epidermal defect in Ikkalpha-/- mice may be due to the failed activation of NF-kappaB. Here we show that the unique function of IKKalpha in control of keratinocyte differentiation is not exerted through its IkappaB kinase activity or through NF-kappaB. Instead, IKKalpha controls production of a soluble factor that induces keratinocyte differentiation.

Effects of polysaccharides from rhizomes of Curcuma zedoaria on macrophage functions.

The effects of Curcuma zedoaria, which is used as a condiment, in perfumery, and as a medicine, on immune response were investigated by measuring macrophage-stimulating activity in macrophages and RAW 264.7 cells. In this study, CZ-1 and CZ-1-III, the fractions partially purified from C. zedoaria, had a strong, dose-dependent lysosomal enzyme activity. It was suggested that active portions of CZ-1-III were polysaccharides rather than proteins. Phagocytic activity increased as a similar pattern in both the gram-negative and gram-positive bacteria, time-dependently. It was demonstrated that CZ-1-III can augment the oxygen burst response but had an even higher activity in vivo than in vitro. Also a significant increase of H2O2, NO, and TNF-alpha production was observed. However, the production of TNF-alpha at the concentration of 1,000 microg/ml decreased. These data suggested that C. zedoaria had macrophage-stimulating activity and the possibility of being used as a biological response modifier.

Heptameric ring structure of the heat-shock protein ClpB, a protein-activated ATPase in Escherichia coli.

The heat-shock protein ClpB is a protein-activated ATPase that is essential for survival of Escherichia coli at high temperatures. ClpB has also recently been suggested to function as a chaperone in reactivation of aggregated proteins. In addition, the clpB gene has been shown to contain two translational initiation sites and therefore encode two polypeptides of different size. To determine the structural organization of ClpB, the ClpB proteins were subjected to chemical cross-linking analysis and electron microscopy. The average images of the ClpB proteins with end-on orientation revealed a seven-membered, ring-shaped structure with a central cavity. Their side-on view showed a two-layered structure with an equal distribution of mass across the equatorial plane of the complex. Since the ClpB subunit has two large regions containing consensus sequences for nucleotide binding, each layer of the ClpB heptamer appears to represent the side projection of one of the major domains arranged on a ring. In the absence of salt and ATP, the ClpB proteins showed a high tendency to form a heptamer. However, they dissociated into various species of oligomers with smaller sizes, depending on salt concentration. Above 0.2 M NaCl, the ClpB proteins behaved most likely as a monomer in the absence of ATP, but assembled into a heptamer in its presence. Furthermore, mutations of the first ATP-binding site, but not the second site, prevented the ATP-dependent oligomerization of the ClpB proteins in the presence of 0.3 M NaCl. These results indicate that ClpB has a heptameric ring-shaped structure with a central cavity and this structural organization requires ATP binding to the first nucleotide-binding site localized to the N-terminal half of the ATPase.

beta-catenin expression and mutational analysis in renal cell carcinomas.

beta-Catenin acts as a downstream transcriptional activator of the Wingless-Wnt signaling pathway. The beta-catenin-Tcf complex transactivates the downstream genes that regulate cell proliferation or inhibit apoptosis. The activation of this pathway through stabilization of beta-catenin is caused either by inactivating mutations of adenomatous polyposis coli (APC) tumor suppressor gene or by activating mutations in beta-catenin exon 3. To determine whether the abnormal expression and activating mutations in exon 3 of the beta-catenin gene are implicated in renal cell carcinogenesis, 52 renal cell carcinomas (RCC) were analyzed by immunohistochemistry, polymerase chain reaction-single-strand conformational polymorphism analysis (PCR-SSCP), and direct DNA sequencing. Immunohistochemically, all cases, as well as normal kidneys, showed membranous and/or cytoplasmic staining patterns without nuclear localization. However, the cytoplasmic accumulations of beta-catenin were observed in five (22.7%) of 22 cases of conventional (clear cell) renal carcinoma, but not in papillary or chromophobe renal carcinomas. The beta-catenin mutation was identified in only one case of conventional renal carcinoma and was a single-base missense mutation on codon 61, leading to substitution of glutamine by arginine. In conclusion, this study demonstrates that beta-catenin mutations are a relatively rare event in RCC and that cytoplasmic accumulations of beta-catenin protein are found only in conventional (clear cell) renal carcinomas. These data suggest that the activation of the beta-catenin signaling pathway may partly play a role in the development of conventional RCC.

Antitumor, genotoxicity and anticlastogenic activities of polysaccharide from Curcuma zedoaria.

The antitumor effect of the partially purified polysaccharide from Curcuma zedoaria was studied in mice transplanted with sarcoma 180 cells. The polysaccharide fraction, CZ-1-III, at dose of 6.25 mg/kg/d showed 50% inhibition in solid tumor growth. When mice were injected with fractions, CZ-1 and CZ-1-III, at the dose of 100.0 mg/kg, 91.6% and 97.1% of tumor growth were inhibited, respectively, indicating that the cytotoxic effect of polysaccharide on sarcoma 180 cells increases upon increasing the amount of polysaccharide administered. To assess the genotoxicity of CZ-1-III fraction, several classical toxicological tests were performed. In Ames test, CZ-1-III did not show any transformation of revertant with or without S-9 metabolic activating system, indicating the lack of mutagenic effect of the compound. To assess clastogenic effect, micronucleus and chromosomal aberration assays were performed using Chinese hamster lung (CHL) fibroblast cells. However, up to 259.0 microg/ml concentration of CZ-1-III, neither micronucleus formation nor chromosomal aberration was induced regardless of the presence of S-9 metabolic activating system. Inhibition of CZ-1-III on micronucleus formation induced by mitomycin C was exhibited in a dose-dependent manner, maximally up to 52.0%. These results strongly suggest that CZ-1-III, the polysaccharide fraction from C. Zedoaria, decreases tumor size of mouse and prevents chromosomal mutation.