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Hepatic transporters, a major determinant of pharmacokinetics, have been used to profile drug properties like efficacy. Among hepatic transporters, importers alter the concentration of the drug by facilitating the transport of a drug into a cell. Despite vast pharmacokinetic studies, the interacting mechanisms of the importers with its substrates or inhibitors are not well understood. Hence, we developed compound binary classification models of whether a compound is binder or nonbinder to a hepatic transporter with experimental data of 284 compounds for four representative hepatic importers, OATP1B1, OATP1B3, OAT2, and OCT1. Support Vector Machine (SVM) along with Genetic Algorithm (GA) was used to construct the classification models of binder versus nonbinder for each target importer. To construct the models, we prepared two data sets, a training data set from Fujitsu database (284 compounds) and an external validation data set from ChEMBL database (1738 compounds). Since an experimental classification criterion between binder and nonbinder has some ambiguity, there is an intrinsic limitation to expect high predictability of the binary classification models developed with the experimental data. The predictability of the classification models calculated with external validation sets were obtained as 77.72%, 84.31%, 84.21%, and 76.38 for OATP1B1, OATP1B3, OAT2, and OCT1, respectively., (Copyright © 2015 The Japanese Society for the Study of Xenobiotics. Published by Elsevier Ltd. All rights reserved.)

Unlabelled: The life cycle of hepatitis C virus (HCV) is highly dependent on host cellular proteins for virus propagation. In order to identify the cellular factors involved in HCV propagation, we performed protein microarray assay using the HCV nonstructural 5A (NS5A) protein as a probe. Of ∼ 9,000 human cellular proteins immobilized in a microarray, approximately 90 cellular proteins were identified as NS5A interactors. Of these candidates, Pim1, a member of serine/threonine kinase family composed of three different isoforms (Pim1, Pim2, and Pim3), was selected for further study. Pim kinases share a consensus sequence which overlaps with kinase activity. Pim kinase activity has been implicated in tumorigenesis. In the present study, we verified the physical interaction between NS5A and Pim1 by both in vitro pulldown and coimmunoprecipitation assays. Pim1 interacted with NS5A through amino acid residues 141 to 180 of Pim1. We demonstrated that protein stability of Pim1 was increased by NS5A protein and this increase was mediated by protein interplay. Small interfering RNA (siRNA)-mediated knockdown or pharmacological inhibition of Pim kinase abrogated HCV propagation. By employing HCV pseudoparticle entry and single-cycle HCV infection assays, we further demonstrated that Pim kinase was involved in HCV entry at a postbinding step. These data suggest that Pim kinase may represent a new host factor for HCV entry., Importance: Pim1 is an oncogenic serine/threonine kinase. HCV NS5A protein physically interacts with Pim1 and contributes to Pim1 protein stability. Since Pim1 protein expression level is upregulated in many cancers, NS5A-mediated protein stability may be associated with HCV pathogenesis. Either gene silencing or chemical inhibition of Pim kinase abrogated HCV propagation in HCV-infected cells. We further showed that Pim kinase was specifically required at an early entry step of the HCV life cycle. Thus, we have identified Pim kinase not only as an HCV cell entry factor but also as a new anti-HCV therapeutic target., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Hepatitis C virus (HCV) usurps host cellular lipid metabolism for production of infectious virus particles. Recently, we have screened a siRNA library targeting host factors that control lipid metabolism and lipid droplet (LD) formation in cell culture grown HCV (HCVcc)-infected cells. Of 10 final candidates, we selected the gene encoding AAM-B for further characterization. We showed that siRNA-mediated knockdown of AAM-B impaired HCV propagation in Jc1-infected cells. More precisely, knockdown of AAM-B abrogated production of infectious HCV particles in both Jc1 RNA electroporated cells and Jc1-infected cells. It is worth noting that knockdown of AAM-B exerted no effect on lipid droplet formation. Moreover, AAM-B interacted with nonstructural 4B (NS4B) through the C-terminal region of NS4B. Protein interplay between AAM-B and NS4B was verified in the context of HCV replication. Using either transient or stable expression of AAM-B, we verified that AAM-B colocalized with NS4B in the cytoplasm. Immunofluorescence data further showed that AAM-B might be involved in recruitment of NS4B to sites in close proximity to LDs to facilitate HCV propagation. Collectively, this study provides new insight into how HCV utilizes cellular AAM-B to facilitate viral propagation.

Purpose: To evaluate the feasibility and safety of robotic thyroidectomy using the da Vinci surgical system., Patients and Methods: Between July 2008 and April 2011, the data revealed an initial series of 100 consecutive patients who underwent robotic thyroidectomy with the da Vinci-S surgical system using the bilateral axillo-breast approach for thyroid cancer. Prospectively collected data were analyzed retrospectively., Results: There were 88 cases of total thyroidectomy, 11 cases of lobectomy, and 1 case of total thyroidectomy with modified radical neck dissection. There was no conversion. The mean total operation time was 287.15±45.19 minutes for total thyroidectomy and 236.27±48.98 minutes for lobectomy. Nineteen patients experienced transient hypocalcemia and 3 patients experienced transient vocal fold palsy. All of the patients recovered within 3 months., Conclusions: Robotic thyroid surgery for patients with thyroid malignancy is safe and results in fewer postoperative complications than open thyroid surgery.

Objective: The purpose of our study was to assess prevalence of carotid intraplaque hemorrhage (IPH) and associations between territorial acute infarction and IPH on magnetization-prepared rapid acquisition with gradient-echo (MPRAGE) in patients with acute neurologic symptoms., Methods: 83 patients with suspected acute neurologic symptoms were evaluated with both brain diffusion weighted imaging (DWI) and carotid MPRAGE sequences. Carotid plaque with high signal intensity on MPRAGE of >200% that of adjacent muscle was categorized as IPH. We analyzed the prevalence of IPH and its correlation with territorial acute infarction., Results: Of 166 arteries, 39 had a carotid artery plaque. Of these arteries, 26 had carotid artery stenosis less than 50%. In all carotid arteries, MR-depicted IPH was found in 7.2% (12/166). High-signal intensity on DWI was found in 17.5% (29/166). Combined lesion with ipsilateral high-signal intensity on DWI and IPH on carotid MPRAGE sequence was found in 6 lesions (6/166, 3.6%). Of patients with carotid artery plaque, MR-predicted IPH was found in 30.8% (12/39) and match lesions with high-signal intensity on DWI and MPRAGE was found in 15.4% (6/39). MR-predicted IPH was significantly higher prevalence in high-grade stenosis group (p=0.010). Relative risk between carotid MPRAGE-positive signal and ipsilateral high-signal intensity on DWI in arteries with carotid artery plaques was 6.8 (p=0.010)., Conclusion: Carotid MPRAGE-positive signal in patients was associated with an increased risk of territorial acute infarction as detected objectively by brain DWI. The relative risk of stroke was increased in high-grade stenosis categories.

Unlabelled: The hepatitis C virus (HCV) life cycle is tightly regulated by lipid metabolism of host cells. In order to identify host factors involved in HCV propagation, we have recently screened a small interfering RNA (siRNA) library targeting host genes that control lipid metabolism and lipid droplet formation using cell culture-grown HCV (HCVcc)-infected cells. We selected and characterized the gene encoding stearoyl coenzyme A (CoA) desaturase 1 (SCD1). siRNA-mediated knockdown or pharmacological inhibition of SCD1 abrogated HCV replication in both subgenomic replicon and Jc1-infected cells, while exogenous supplementation of either oleate or palmitoleate, products of SCD1 activity, resurrected HCV replication in SCD1 knockdown cells. SCD1 was coimmunoprecipitated with HCV nonstructural proteins and colocalized with both double-stranded RNA (dsRNA) and HCV nonstructural proteins, indicating that SCD1 is associated with HCV replication complex. Moreover, SCD1 was fractionated and enriched with HCV nonstructural proteins at detergent-resistant membrane. Electron microscopy data showed that SCD1 is required for NS4B-mediated intracellular membrane rearrangement. These data further support the idea that SCD1 is associated with HCV replication complex and that its products may contribute to the proper formation and maintenance of membranous web structures in HCV replication complex. Collectively, these data suggest that manipulation of SCD1 activity may represent a novel host-targeted antiviral strategy for the treatment of HCV infection., Importance: Stearoyl coenzyme A (CoA) desaturase 1 (SCD1), a liver-specific enzyme, regulates hepatitis C virus (HCV) replication through its enzyme activity. HCV nonstructural proteins are associated with SCD1 at detergent-resistant membranes, and SCD1 is enriched on the lipid raft by HCV infection. Therein, SCD1 supports NS4B-mediated membrane rearrangement to provide a suitable microenvironment for HCV replication. We demonstrated that either genetic or chemical knockdown of SCD1 abrogated HCV replication in both replicon cells and HCV-infected cells. These findings provide novel mechanistic insights into the roles of SCD1 in HCV replication., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Viruses exploit cellular SUMOylation machinery to favour their own propagation. We show that NS5A is a target protein of small ubiquitin-like modifier (SUMO) and is SUMOylated at lysine residue 348. We demonstrated that SUMOylation increased protein stability of NS5A by inhibiting ubiquitylation, and SUMOylation was also required for protein interaction with NS5B. These data imply that SUMO modification may contribute to HCV replication. Indeed, silencing of UBC9 impaired HCV replication in Jc1-infected cells, and HCV replication level was also significantly reduced in SUMO-defective subgenomic replicon cells. Taken together, these data indicate that HCV replication is regulated by SUMO modification of NS5A protein. We provide evidence for the first time that HCV exploits host cellular SUMO modification system to favour its own replication., (© 2014 John Wiley & Sons Ltd.)

Spontaneous intracranial vertebrobasilar dissection can manifest with various clinical symptoms, including subarachnoid hemorrhage or ischemic symptoms from impaired posterior circulation. A 29-year-old woman came to our emergency department with a sudden onset of left sided mild motor weakness and headache. Initial magnetic resonance imaging (MRI) showed mild luminal irregularities in the vertebrobasilar arteries with an eccentric periluminal hematoma. Follow-up MRI obtained 3 days later showed a progression of vertebrobasilar dissection to multifocal stenoses with an increased intramural hematoma.

In a previous study, we constructed a male reference Korean phantom; HDRK-Man (High-Definition Reference Korean-Man), to represent Korean adult males for radiation protection purposes. In the present study, a female phantom; HDRK-Woman (High-Definition Reference Korean-Woman), was constructed to represent Korean adult females. High-resolution color photographic images obtained by serial sectioning of a 26 year-old Korean adult female cadaver were utilized. The body height and weight, the skeletal mass, and the dimensions of the individual organs and tissues were adjusted to the reference Korean data. The phantom was then compared with the International Commission on Radiological Protection (ICRP) female reference phantom in terms of calculated organ doses and organ-depth distributions. Additionally, the effective doses were calculated using both the HDRK-Man and HDRK-Woman phantoms, and the values were compared with those of the ICRP reference phantoms.

To identify the novel genes involved in lipid metabolism and lipid droplet formation that may play important roles in Hepatitis C virus (HCV) propagation, we have screened the small interfering RNA library using cell culture derived HCV (HCVcc)-infected cells. We selected and characterized the gene encoding farnesyl-diphosphate farnesyltransferase 1 (FDFT1). siRNA-mediated knockdown of FDFT1 impaired HCV replication in both subgenomic replicon and HCVcc infected cells. Moreover, YM-53601, an inhibitor of FDFT1 enzyme activity, abrogated HCV propagation. HCV infection increased FDFT1 protein level but not FDFT1 mRNA level. These results suggest that HCV may modulate FDFT1 protein level to facilitate its own propagation., (Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

Background: Endoscopic thyroid surgery is gaining wide acceptance; however, existing endoscopic methods for thyroidectomy have shown several limitations. Recently, a transoral technique using video assistance and endoscopy has been reported for thyroidectomy. The aim of this study was to define a new technique of transoral thyroidectomy using a mandibular periosteal approach to complement other types of natural orifice surgery and minimally invasive surgery., Materials and Methods: Transoral periosteal thyroidectomies were performed in seven living pigs to evaluate the feasibility and safety of the new approach. Total thyroidectomies were performed in all animals. Follow-up examinations were carried out for 7 days and followed by autopsy., Results: Through three trocars in the mandibular periosteal area, it was possible to create a working space under the platysma muscle and to reach the pretracheal area. Total thyroidectomies were also performed via the transoral, mandibular periosteal approach without complications in seven orally intubated living pigs. Postoperatively, the white blood cell count remained normal in all cases. On the postoperative sacrifice of the pigs, three locally encapsulated seromas were observed. Both recurrent laryngeal nerves were intact in all cases., Conclusions: Transoral periosteal thyroidectomy could be feasible and safe.

Dissection of the middle cerebral artery (MCA) is less frequent compared with dissection of vessels in the vertebrobasilar system or dissection of the carotid artery. High-resolution cross-sectional magnetic resonance imaging (HRMRI) has emerged as a potential technique for atherosclerotic plaque imaging in MCA. We enrolled 3 patients with MCA dissection on whom HRMRI was performed for evaluation of MCA stenosis. Two patients had an embolic infarction in the MCA territory and focal dissection. One patient had a massive infarction in the MCA territory and long-segment dissection of the MCA. On HRMRI, our objectives had an intimal flap with patency of the lumen and 1 had extensive hemorrhaging in the false lumen., (Copyright © 2014. Published by Elsevier Inc.)

Purpose: To evaluate the safety and efficacy of emergency carotid stenting and recanalization using the modified Penumbra System (PS) technique for cases of acute extracranial internal carotid artery (ICA) occlusion and massive thrombus with acute ischemic stroke symptoms., Methods: The clinical outcomes of patients with tandem occlusion of the ICA and middle cerebral artery (MCA) who underwent recanalization by a modified PS technique after carotid artery stenting during the period December 2009 to June 2011 were reviewed., Results: Five patients with tandem occlusion of the ICA and MCA were enrolled. The mean National Institute of Health Stroke Scale (NIHSS) score on admission was 13 (range 9-18). The median time from symptom onset to arrival at hospital was 260 min. The median time lag between arrival at hospital and intra-arterial administration was 70 min. Recanalization by stenting of carotid artery occlusion was successfully performed in all patients. The mean number of recanalizations using the modified PS technique for distally migrated thrombus was 3. Complete recanalization of the MCA was observed in all patients. Symptomatic hemorrhage transformation did not occur during the hospital stay in any of the subjects. At discharge, the mean NIHSS score was 2.2 (range 1-6). At the 3-month follow-up the modified Rankin Scale was excellent in three of the five patients, good in one patient and poor in one patient., Conclusions: The modified PS technique after carotid artery stenting is a safe and effective treatment for patients with a massive thrombus extending into the distal portion of the ICA and a tandem occlusion of the ICA and MCA.

In the Visible Korean project, serially sectioned images of the pelvis were made from a female cadaver. Outlines of significant structures in the sectioned images were drawn and stacked to build surface models. To improve the accessibility and informational content of these data, a five-step process was designed and implemented. First, 154 pelvic structures were outlined with additional surface reconstruction to prepare the image data. Second, the sectioned and outlined images (in a browsing software) as well as the surface models (in a PDF file) were placed on the Visible Korean homepage in a readily-accessible format. Third, all image data were visualized with interactive elements to stimulate creative learning. Fourth, two-dimensional (2D) images and three-dimensional (3D) models were superimposed on one another to provide context and spatial information for students viewing these data. Fifth, images were designed such that structure names would be shown when the mouse pointer hovered over the 2D images or the 3D models. The state-of-the-art sectioned images, outlined images, and surface models, arranged and systematized as described in this study, will aid students in understanding the anatomy of female pelvis. The graphic data accompanied by corresponding magnetic resonance images and computed tomographs are expected to promote the production of 3D simulators for clinical practice., (Copyright © 2013 American Association of Anatomists.)

Hepatitis C virus (HCV) is highly dependent on cellular factors for its own propagation. By employing tandem affinity purification method, we identified pyruvate carboxylase (PC) as a cellular partner for NS5A protein. NS5A interacted with PC through the N-terminal region of NS5A and the biotin carboxylase domain of PC. PC expression was decreased in cells expressing NS5A and HCV-infected cells. Promoter activity of PC was also decreased by NS5A protein. However, FAS expression was increased in cells expressing NS5A and cell culture grown HCV (HCVcc)-infected cells. Silencing of PC promoted fatty acid synthase (FAS) expression level. These data suggest HCV may modulate PC via NS5A protein for its own propagation.

Spindle cell/pleomorphic lipoma is an uncommon benign adipose tissue tumor most frequently arising from the subcutaneous tissue of the back, shoulder, head and neck, and extremities. The deep cervical spaces are the rarely affected locations. Herein we report on the imaging findings of spindle cell/pleomorphic lipoma involving the retropharyngeal space in an elderly woman.

Hepatitis C virus (HCV) is highly dependent on cellular proteins for its own propagation. In order to identify the cellular factors involved in HCV propagation, we performed protein microarray assays using the HCV core protein as a probe. Of ~9,000 host proteins immobilized in a microarray, approximately 100 cellular proteins were identified as HCV core-interacting partners. Of these candidates, mitogen-activated protein kinase-activated protein kinase 3 (MAPKAPK3) was selected for further characterization. MAPKAPK3 is a serine/threonine protein kinase that is activated by stress and growth inducers. Binding of HCV core to MAPKAPK3 was confirmed by in vitro pulldown assay and further verified by coimmunoprecipitation assay. HCV core protein interacted with MAPKAPK3 through amino acid residues 41 to 75 of core and the N-terminal half of kinase domain of MAPKAPK3. In addition, both RNA and protein levels of MAPKAPK3 were elevated in both HCV subgenomic replicon cells and cell culture-derived HCV (HCVcc)-infected cells. Silencing of MAPKAPK3 expression resulted in decreases in both protein and HCV infectivity levels but not in the intracellular HCV RNA level. We showed that MAPKAPK3 increased HCV IRES-mediated translation and MAPKAPK3-dependent HCV IRES activity was further increased by core protein. These data suggest that HCV core may modulate MAPKAPK3 to facilitate its own propagation.

Background and Purpose: Patients who develop severe stroke symptoms due to acute internal carotid artery occlusion eventually in combination with a thromboembolic obstruction of the middle cerebral artery incur a major risk of developing extensive MCA infarction with a poor outcome. The purpose of this study was to evaluate the outcome for patients with tandem occlusions in the MCA and/or distal ICA, retrospectively, who had undergone stent implantation in the proximal segment of the ICA in addition to intra-arterial thrombolysis., Materials and Methods: Thirty-five patients with tandem occlusions of the MCA and/or distal ICA and acute occlusion of the proximal ICA underwent stent implantation for the proximal ICA occlusion and IAT for the tandem occlusion. Clinical outcome measures were assessed on admission and at discharge by using the National Institutes of Health Stroke Scale as well as 3 months after treatment by using the modified Rankin Scale., Results: The median NIHSS score on admission was 12 (range, 6-22). All patients had patent flow into the M1 and ICA after carotid artery stent placement and IAT. After the procedure, 19 patients (54.3%) were TICI grade III; 7 (20.0%), TICI grade IIb; and 9 (25.7%), TICI grade IIa. Symptomatic intracerebral hemorrhage occurred in 1 patient (2.9%). The overall mortality rate was 11.4% (4/35). At 3-month follow-up, the median NIHSS score was 4 (range, 1-17). NIHSS score at admission and TICI grade were all found to be independently associated with an unfavorable outcome at 3 months., Conclusions: Initial stroke severity, degree of successful revascularization, and the side of ischemia were found to independently predict the functional outcome at 3 months after treatment.

Objective: The ocular vestibular-evoked myogenic potential (oVEMP), a recently documented otolith-ocular reflex, is considered to reflect the central projections of the primary otolithic afferent fibers to the oculomotor nuclei. The aim of our study is to define air-conducted sound oVEMP abnormality in patients with acute brainstem lesions and to determine the brainstem structures involved in the generation of oVEMPs., Methods: In response to air-conducted tone burst sounds (ACS), oVEMP was measured in 52 patients with acute brainstem lesions. Individualized brainstem lesions were analyzed by means of MRI-based voxel-wise lesion-behavior mapping, and the probabilistic lesion maps were constructed., Results: More than half (n=28, 53.8%) of the patients with acute brainstem lesions showed abnormal oVEMP in response to ACS. The majority of patients with abnormal oVEMPs had lesions in the dorsomedial brainstem that contains the medial longitudinal fasciculus (MLF), the crossed ventral tegmental tract (CVTT), and the oculomotor nuclei and nerves., Conclusion: MLF, CVTT, and the oculomotor nuclei and nerves appear to be responsible for otolith-ocular responses in the brainstem., Significance: Complemented to cervical VEMP for the uncrossed otolith-spinal function, oVEMP to ACS may be applied to evaluate the crossed otolith-ocular function in central vestibulopathies., (Copyright © 2012 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.)

Myoid angioendothelioma of the spleen is an uncommon, benign vascular tumor that is morphologically characterized by a composite of vascular spaces and stromal cells with myoid feature. Herein, we report a case of the myoid angioendothelioma of the spleen, concurrent with rectal adenocarcinoma. A 41-year-old woman presented with hematochezia for several weeks. Grossly, the rectal mass was a 2.5 × 2-cm ulcerative fungating lesion. The splenic mass was a 2.2 × 2-cm well-circumscribed lesion. Microscopically, the rectal mass was a well-differentiated adenocarcinoma that invaded into the pericolic adipose tissue. The splenic mass was composed of slit-like vascular spaces and fascicles of elongated stromal cells. Vascular endothelial cells were immunopositive for CD31, factor VIII-related antigen, and CD34 but negative for CD8. Stromal cells were immunopositive for smooth muscle actin but negative for desmin., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The latent ribonuclease RNase L and the interferon-inducible 2',5'-oligoadenylate synthetase (OAS) have been implicated in the antiviral response against hepatitis C virus (HCV). However, the specific roles of these enzymes against HCV have not been fully elucidated. In this study, a scarce endogenous expression and RNA degrading activity of RNase L in human hepatoma Huh7 cells enabled us to demonstrate the antiviral activity of RNase L against HCV replication through the transient expression of the enzyme. The antiviral potential of specific members of the OAS family was further examined through overexpression and RNA interference approaches. Our data suggested that among the members of the OAS family, OAS1 p46 and OAS3 p100 mediate the RNase L-dependent antiviral activity against HCV., (Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

Background: Preservation of the external branch of the superior laryngeal nerve (EBSLN) during thyroidectomy is important because its injury may lead to frequent occurrence of vocal fatigue and the inability to perform superenergetic phonation. Most studies on the anatomy of EBSLN have been performed in Western countries and, thus, have some limitations in their application to Koreans. We explored the anatomy of EBSLNs using intraoperative neuromonitoring (IONM) in Korean adults., Methods: Between August 2011 and December 2011, 50 patients underwent thyroidectomy at Korea University, Anam Hospital in Seoul, Korea. IONM was performed with the NIM response 3.0 system., Results: Forty-two total thyroidectomies and eight lobectomies were performed with IONM and 92 EBSLNs were evaluated. Type I EBSLN was observed in 15 of the 92 nerves (16.3%), type IIa EBSLN was noted in 52 (56.5%) and type IIb EBSLN was noted in 25 (27.2%). Patients with types IIa and IIb were at higher risk of injuries and these types were more frequently observed (83.7%) compared with previous Western studies. We found that 35.9% of distal insertion sites of EBSLNs were located within 1cm of the center of the cricoid cartilage., Conclusion: Surgeons should pay close attention to preserving the EBSLN during thyroidectomy in Korean patients because their EBSLNs are more frequently located beneath the superior thyroid vessels., (Copyright © 2012. Published by Elsevier B.V.)

Hepatitis C virus (HCV) infection is a major cause of liver disease, including cirrhosis and hepatocellular carcinoma. Due to significant adverse effects and emergence of resistant strains of currently developed anti-HCV agents, plant extracts have been considered to be potential sources of new bioactive compounds against HCV. The aim of this study was to evaluate the functional effects of triterpenoid saponins contained in the root extract of Platycodon grandiflorum (PG) on viral enzyme activities and replication in both HCV replicon cells and cell culture grown HCV- (HCVcc-) infected cells. Inhibitory activities of triterpenoid saponins from PG were verified by NS5B RNA-dependent RNA polymerase assay and were further confirmed in the context of HCV replication. Six triterpenoid saponins (platycodin D, platycodin D2, platycodin D3, deapioplatycodin D, deapioplatycodin D2, and platyconic acid A), PG saponin mixture (PGSM), were identified as active components exerting anti-HCV activity. Importantly, PGSM exerted synergistic anti-HCV activity in combination with either interferon- α or NS5A inhibitors. We demonstrated that combinatorial treatment of PGSM and IFN- α efficiently suppressed colony formation with significant reduction in drug resistant variant of HCV. These data suggest that triterpenoid saponin may represent a novel anti-HCV therapeutic agent.

Purpose: High resolution magnetic resonance imaging (HRMRI) has been used as an imaging modality to depict the intracranial artery wall. The aim of this study was to compare images of the vessel wall between symptomatic and asymptomatic atherosclerotic plaques of the middle cerebral artery (MCA) using HRMRI., Materials and Methods: From September 2009 to August 2010 we prospectively screened consecutive patients for MCA stenosis using time-of-flight (TOF) MR angiography. We studied 14 patients with symptomatic MCA stenosis and 16 patients with asymptomatic MCA stenosis. The HRMRI protocol included three different scans: T1-, T2-, and proton density (PD)-weighted black blood MRI. The cross-sectional images of the MCA wall on HRMRI were compared between the two groups based on the degree of stenosis, remodeling ratio, outward or inward remodeling, plaque signal intensity, plaque surface irregularity, and presence of an intact inner wall., Results: The degree of MCA stenosis and the ratio of plaque thickening to patent lumen in the symptomatic group were significantly higher than in the asymptomatic group. Outward remodeling of the stenotic area in symptomatic group was significantly higher than that seen in the asymptomatic group, and the reverse was true for inward remodeling of the stenotic area (it was significantly higher in the asymptomatic group compared to the symptomatic group). T2- and PD-weighted high signal foci, eccentric wall thickening, and plaque volume in the stenotic area were all similar between the two groups., Conclusions: HRMRI has the potential to distinguish between atherosclerotic plaques in symptomatic and asymptomatic MCA stenoses., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Central pontine myelinolysis is a rare neurologic disorder that is defined by demyelination of longitudinally descending tracts and transversly crossing fibers in the basis pontis. Frequently observed clinical manifestations of this disorder include sudden weakness, dysphagia, loss of consciouness and locked-in syndrome. However, there have been a few studies that reported a benign course of this disease, which include cerebellar signs, such as ataxia, intention tremor, and dysarthria. Here we report on a 53-year-old male with a history of liver cirrhosis who showed the cerebellar type of central pontine myelinolysis. The patient was diagnosed with central pontine myelinolysis based on clinical presentations and magnetic resonance imaging findings after a liver transplantation. Conventional magenetic resonance imaging (MRI) revealed the preservation of the corticospinal tract and abnormal pontocerebellar fibers. However, these findings were not sufficient to define the pathophysiology of our patient. Electrophysiologic analysis and diffusion tensor imaging (DTI) were performed to investigate cerebellar signs in this case. Delayed central motor conduction time (CMCT) to the tibialis anterior muscle with transcranial magnetic stimulation (TMS) was observed, which indicated demyelination of the corticospinal tract. Also, diffusion tensor imaging showed abnormal pontocerebellar fibers, which might have been caused by cerebellar dysfunction in our patient. A combination of TMS and DTI was also used to determine the pathophysiology of this disease.

Background & Aims: Hepatitis C virus (HCV) requires host cellular proteins for its own propagation. To identify the cellular factors necessary for HCV propagation, we have recently screened the small interfering RNA (siRNA) library targeting cell cycle genes using cell culture grown HCV (HCVcc)-infected cells. In the current study, we have selected and characterized the gene encoding Cyclin A2 (CycA2). Deregulation of CycA2 has been implicated in many types of cancers, including hepatocellular carcinoma., Methods: The effects of CycA2 on HCV propagation were investigated by siRNA-mediated knockdown assay, in vitro and in vivo protein binding assays, luciferase reporter gene assay, and immunoblot assay., Results: We showed that siRNA-mediated depletion of CycA2 significantly inhibited HCV replication in both HCV subgenomic replicon cells and HCVcc-infected cells. Furthermore, HCV non-structural 5B (NS5B) specifically interacted with CycA2 in vitro and in vivo. Protein interaction was mediated through the cyclin box of CycA2 and the palm domain of NS5B. We further showed that R/HxL motif in the palm domain of HCV NS5B mediated protein interaction with CycA2 and this interaction was necessary for HCV replication. Moreover, we demonstrated that tylophorine, the natural plant product exerting a CycA2 inhibitory function, abrogated HCV replication., Conclusions: HCV regulates CycA2 via NS5B protein for its own propagation. In addition, tylophorine may be a potential therapeutic agent for HCV., (Copyright © 2012 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.)

We identified heat shock protein 72 (Hsp72) as a host factor that was differentially expressed in cells expressing nonstructural 5A (NS5A) protein. To investigate how NS5A modulates Hsp72 in hepatitis C virus (HCV) life cycle, we examined the role of Hsp72 in HCV replication and virus production. NS5A specifically interacted with Hsp72. Both Hsp72 and nuclear factor of activated T cells 5 (NFAT5) levels were increased in cells expressing NS5A protein. Treatments of N-acetylcysteine and glutathione markedly reduced protein levels of both NFAT5 and Hsp72. Knockdown of NFAT5 resulted in decrease in Hsp72 level in cells expressing NS5A. Importantly, silencing of Hsp72 expression resulted in decrease in both RNA replication and virus production in HCV-infected cells. These data indicate that NS5A modulates Hsp72 via NFAT5 and reactive oxygen species activation for HCV propagation., (© 2011 Blackwell Publishing Ltd.)

Purpose: Sectioned images of cadavers enable creation of realistic three-dimensional (3D) models. In order to build a 3D model of a structure, the structure has to be outlined in the sectioned images. The outlining process is time consuming; therefore, users want to be provided with outlined images. The more detailed structures are outlined, the greater potential for wider application of the outlined images., Methods: In the Visible Korean, sectioned images (intervals 0.2 mm) of the entire body of a male cadaver were prepared. In the available 1,702 sectioned images (intervals 1 mm), 937 structures were outlined over a period of 8 years. The outlined images were altered to black-filled images for each structure; black-filled images were selected for distribution in order to maintain small file sizes., Results: We attempted to determine whether black-filled images could be used in various situations. The outlines of these images were interpolated for production of new images at 0.2 mm intervals. The outlines were then filled with different colors for construction of color-filled images of all structures. Volume and surface reconstructions of the black-filled images were executed in order to build volume and surface models., Conclusion: The black-filled images with corresponding sectioned images presented here are the source of realistic 3D models for use in medical simulation systems.

Purpose: Circulating tumor cells (CTC) clearly correlate with unfavorable outcomes for patients with metastatic breast cancer, but the long-term prognostic implications of CTC for molecular subtypes of operable breast cancer are not yet known. We explored the relationships between previously established prognostic factors and CTC in operable breast cancer, and the significance of CTC by breast cancer molecular subtype., Methods: We retrospectively evaluated 166 patients with operable breast cancer (stage I-IIIA) diagnosed from April 1997 to May 2003. CTC were detected using cytokeratin-20 (CK-20) mRNA expression in peripheral blood samples that were collected just prior to surgery under general anesthesia. Clinicopathological characteristics of the cancer were analyzed according to CTC status. Metastasis-free survival (MFS) and overall survival (OS) were analyzed according to CTC status and breast cancer molecular subtype., Results: CK-20 mRNA-positive CTC was detected in 37 of 166 patients (22.3%) and was not correlated with any previous clinical factors in univariate analysis (p>0.05). After a median follow-up of 100 months, the patients with CK-20 mRNA-positive CTC had less favorable outcomes in terms of MFS and OS than those without detectable CTC (log-rank p<0.05). Among molecular subtypes of operable breast cancer, the patients with CK-20 mRNA-positive CTC had shorter MFS and OS in triple negative and human epidermal growth factor 2 (HER2) breast cancer subtype (log-rank, p<0.05)., Conclusion: CK-20 mRNA-positive CTC may lend insight into tumor progression as a prognostic indicator especially in the triple negative and HER2 subtypes of operable breast cancer.

We present the results of our approach for treating 12 consecutive cases of acute middle cerebral artery (MCA) stroke by performing balloon-expandable stent (BES) placement after immediate reocclusion due to the underlying stenosis after intra-arterial thrombolysis (IAT). We retrospectively reviewed the clinical outcomes of 12 patients with acute MCA stroke who underwent recanalization by BES placement in an underlying stenosis after IAT. The time to treatment, urokinase dose, duration of the procedure, recanalization rates and symptomatic hemorrhage were analyzed. Clinical outcome measures were assessed on admission and at discharge (the National Institutes of Health stroke scores [NIHSS]) as well as three months after treatment (modified Rankin scales [mRS]). The median NIHSS score on admission was 8.6. Four patients received IV rtPA. The median time from symptom onset to IAT was 236 minutes and the median duration of IAT was 62 minutes. The median dose of urokinase was 140,000 units. Initial recanalization after stent deployment (thrombolysis in cerebral ischemia attack grade of II or III) was achieved in all patients. Two patients died in the hospital due to aspiration pneumonia during medical management. In two patients, in-stent reocclusion occurred within 48 hours after stent deployment. At discharge, the median NIHSS score in ten patients (including the patients with reobstruction) was 2.4. The three-month outcome was excellent (mRS, 0-1) in eight patients. In this study, BES deployment was safe and effective in patients with an immediately reoccluded MCA after successful IAT.

Saponins are a group of naturally occurring plant glycosides which possess a wide range of pharmacological properties, including anti-tumorigenic and antiviral activities. To investigate whether saponin has anti-hepatitis C virus (HCV) activity, we examined the effect of saponin on HCV replication. HCV replication was efficiently inhibited at a concentration of 10 µg/ml of saponin in cell culture grown HCV (HCVcc)-infected cells. Inhibitory effect of saponin on HCV replication was verified by quantitative real-time PCR, reporter assay, and immunoblot analysis. In addition, saponin potentiated IFN-α-induced anti-HCV activity. Moreover, saponin exerted antiviral activity even in IFN-α resistant mutant HCVcc-infected cells. To investigate how cellular genes were regulated by saponin, we performed microarray analysis using HCVcc-infected cells. We demonstrated that suppressor of cytokine signaling 2 (SOCS2) protein level was distinctively increased by saponin, which in turn resulted in inhibition of HCV replication. We further showed that silencing of SOCS2 resurrected HCV replication and overexpression of SOCS2 suppressed HCV replication. These data imply that saponin inhibits HCV replication via SOCS2 signaling pathway. These findings suggest that saponin may be a potent therapeutic agent for HCV patients.

Mechanical clot disruption for the treatment of acute basilar artery occlusion (BAO) is known to provide a benefit. We aimed to determine the safety, recanalization rate and time-to-flow restoration of mechanical clot disruption and low dose urokinase (UK) infusions for the treatment of patients with acute BAO. Between June 2006 and June 2010, 21 patients with acute BAO underwent endovascular treatment that included angioplasty or stent placement. The time to treatment, duration of the procedure, dose of urokinase (UK), recanalization rates and symptomatic hemorrhages were analyzed. Clinical outcome measures were assessed at admission and at the time of discharge using the National Institutes of Health Stroke Scale (NIHSS) score and at three months after treatment using the modified Rankin Score (mRS). On admission, the median NIHSS score was 13.2. Median time from symptom onset to arrival at hospital was 356 minutes, and median time from symptom onset to intraarterial thrombolysis (IAT) was 49 minutes. We used the following interventional treatment regimens: Intra-arterial (IA) UK and a minimal mechanical procedure (n=14), IA UK with angioplasty (n=1), IA UK with angioplasty and stent placement (n=3) and IA UK with HyperForm (n=3). The recanalization (thrombolysis in cerebral ischemia grade II or III) rate was 90.5% (19/21). There was symptomatic hemorrhage in one patient (4.8%). The median NIHSS score at discharge was 6.3. The three-month outcome was favorable (mRS: 0-2) for 14 patients (66.7%) and poor (mRS: 3-6) for seven patients (33.3%). The overall mortality at three months was 14.3% (three patients died). Low-dose IAT with mechanical clot disruption is a safe and effective treatment for treatment for acute BAO.

The interpretation of computed tomographs (CTs) and magnetic resonance images (MRIs) to diagnose clinical conditions requires basic knowledge of sectional anatomy. Sectional anatomy has traditionally been taught using sectioned cadavers, atlases, and/or computer software. The computer software commonly used for this subject is practical and efficient for students but could be more advanced. The objective of this research was to present browsing software developed from the Visible Korean images that can be used for teaching sectional anatomy. One thousand seven hundred and two sets of MRIs, CTs, and sectioned images (intervals, one millimeter) of a whole male cadaver were prepared. Over 900 structures in the sectioned images were outlined and then filled with different colors to elaborate each structure. Software was developed where four corresponding images could be displayed simultaneously; in addition, the structures in the image data could be readily recognized with the aid of the color-filled outlines. The software, distributed free of charge, could be a valuable tool to teach medical students. For example, sectional anatomy could be taught by showing the sectioned images with real color and high resolution. Students could then review the lecture by using the sectioned and color-filled images on their own computers. Students could also be evaluated using the same software. Furthermore, other investigators would be able to replace the images for more comprehensive sectional anatomy., (Copyright © 2011 Wiley-Liss, Inc.)

Hepatitis C virus (HCV) RNA replication requires cellular factors as well as viral non-structural proteins (NS protein). Using small interfering RNA (siRNA) library screening, we previously identified c-Fos as a host factor involved in HCV propagation. In the present study, we demonstrated that silencing of c-Fos expression resulted in decrease of HCV propagation in cell culture grown HCV (HCVcc)-infected cells; whereas overexpression of c-Fos significantly increased HCV propagation. We further confirmed the positive role of c-Fos in HCV propagation by both HCV-luciferase reporter assay and immunofluorescence analysis. We showed that c-Fos level was upregulated by HCV infection. Furthermore, phorbol 12-myristate 13-acetate (PMA)-induced c-Fos level was synergistically increased by HCV infection. These data suggest that c-Fos acts as a positive regulator of HCV propagation and may contribute to HCV-associated pathogenesis., (Copyright © 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

The life cycle of hepatitis C virus (HCV) is highly dependent on cellular factors. Using small interfering RNA (siRNA) library screening, we identified peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (Pin1) as a host factor involved in HCV propagation. Here we demonstrated that silencing of Pin1 expression resulted in decreases in HCV replication in both HCV replicon cells and cell culture-grown HCV (HCVcc)-infected cells, whereas overexpression of Pin1 increased HCV replication. Pin1 interacted with both the NS5A and NS5B proteins. However, Pin1 expression was increased only by the NS5B protein. Both the protein binding and isomerase activities of Pin1 were required for HCV replication. Juglone, a natural inhibitor of Pin1, inhibited HCV propagation by inhibiting the interplay between the Pin1 and HCV NS5A/NS5B proteins. These data indicate that Pin1 modulates HCV propagation and may contribute to HCV-induced liver pathogenesis.

Purpose: The aim of this study was to evaluate the usefulness of three-dimensional CT angiography (3D CTA) with bone subtraction in a comparison with 3D CTA without bone subtraction for the detection of intracranial aneurysms., Materials and Methods: Among 337 consecutive patients who had intracranial aneurysms detected on 3D CTA, 170 patients who underwent digital subtraction angiography (DSA) were included in the study. CTA was performed with a 16-slice multidetector-row CT (MDCT) scanner. We created the 3D reconstruction images with and without bone subtraction by using the volume rendering technique. Three neuroradiologists in a blinded fashion interpreted both 3D CTA images with and without bone subtraction. The diagnostic accuracy of both techniques was evaluated using the alternative free-response receiver operating characteristic (ROC) analysis. The sensitivity and positive predictive value were also evaluated., Results: A total of 200 aneurysms (size: 2-23 mm) were detected in 170 patients. The area under the receiver operating characteristic curve (Az) for 3D CTA with bone subtraction (mean, Az=0.933) was significantly higher than that for 3D CTA without bone subtraction (mean, Az=0.879) for all observers (P<0.05). The sensitivity of 3D CTA with bone subtraction for three observers was 90.0, 92.0 and 92.5%, respectively, while the sensitivity of 3D CTA without bone subtraction was 83.5, 83.5 and 87.5%, respectively. No significant difference in positive predictive value was observed between the two modalities., Conclusions: 3D CTA with bone subtraction showed significantly higher diagnostic accuracy for the detection of intracranial aneurysms as compared to 3D CTA without bone subtraction., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Even though the hybrid phantom embodies both the anatomic reality of voxel phantoms and the deformability of stylized phantoms, it must be voxelized to be used in a Monte Carlo code for dose calculation or some imaging simulation, which incurs the inherent limitations of voxel phantoms. In the present study, a voxel phantom named VKH-Man (Visible Korean Human-Man), was converted to a polygon-surface phantom (PSRK-Man, Polygon-Surface Reference Korean-Man), which was then adjusted to the Reference Korean data. Subsequently, the PSRK-Man polygon phantom was directly, without any voxelization process, implemented in the Geant4 Monte Carlo code for dose calculations. The calculated dose values and computation time were then compared with those of HDRK-Man (High Definition Reference Korean-Man), a corresponding voxel phantom adjusted to the same Reference Korean data from the same VKH-Man voxel phantom. Our results showed that the calculated dose values of the PSRK-Man surface phantom agreed well with those of the HDRK-Man voxel phantom. The calculation speed for the PSRK-Man polygon phantom though was 70-150 times slower than that of the HDRK-Man voxel phantom; that speed, however, could be acceptable in some applications, in that direct use of the surface phantom PSRK-Man in Geant4 does not require a separate voxelization process. Computing speed can be enhanced, in future, either by optimizing the Monte Carlo transport kernel for the polygon surfaces or by using modern computing technologies such as grid computing and general-purpose computing on graphics processing units programming.

Hepatitis C Virus (HCV) nonstructural 5A (NS5A) is a pleiotropic protein involved in viral RNA replication and modulation of the cellular physiology in HCV-infected cells. To elucidate the mechanisms of the HCV life cycle, we identified cellular factors interacting with the NS5A protein in HCV-infected cells. Huh7.5 cells were electroporated with HCV Jc1 RNA. Cellular factors associated with HCV NS5A were identified by immunoprecipitation with Dynabead-conjugated NS5A antibody and LC-MS/MS. Phosphatidylinositol 4-kinase type IIIα (PI4KIIIα) was identified as a binding partner for the NS5A protein. NS5A derived from both genotypes 1b and 2a interacted with PI4KIIIα. NS5A interacted with PI4KIIIα through amino acids 401-600 of PI4KIIIα and domain I of NS5A. Interference of the protein interaction between NS5A and PI4KIIIα decreased HCV propagation. Knockdown of PI4KIIIα significantly reduced HCV replication in Huh7 cells harboring the subgenomic replicon and in Huh7.5 cells infected with cell culture grown virus (HCVcc). Silencing of PI4KIIIα further inhibited HCV release into the tissue culture medium. NS5A may recruit PI4KIIIα to the HCV RNA replication complex. These data suggest that PI4KIIIα is an essential host factor that supports HCV proliferation and therefore PI4KIIIα may be a legitimate target for anti-HCV therapy.

To investigate the molecular mechanisms underlying interferon alpha (IFNα) treatment failure in hepatitis C virus (HCV) patients with chronic hepatitis, we aimed to develop an IFNα-resistant clone of HCV. By treating JFH-1-infected Huh7.5 cells with a prolonged low-dose treatment of IFNα, we selected a clone of HCV that survived against 100 U/ml of IFNα. By genetic analysis of this clone, we found four substitution mutations in the C-terminal coding sequence of non-structural 5A (NS5A). By introducing these four mutations into wild-type JFH-1, we established a new HCV clone that acquired IFNα resistant phenotype. These data suggest that four amino acid substitutions in NS5A are involved in IFNα resistance and thus this newly established HCV may be a useful tool for elucidating the molecular mechanisms of IFNα resistance in HCV patients., (Copyright © 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

This study compared the efficacy of gadoxetic acid-enhanced MRI and gadopentetate dimeglumine-enhanced MRI in the detection of small hepatocellular carcinoma (HCC). Both MRI techniques were performed on 43 patients with a total of 59 HCCs (size range, 0.5-2.0 cm), with a mean interval between the two MRI studies of 3 days (range, 2-7 days). Two observers reviewed both data sets in consensus. Diagnostic accuracy and sensitivity were evaluated using the alternative-free response receiver operator characteristic (ROC) method. The gadoxetic acid set showed a trend toward increased area under the ROC curve (Az value = 0.958) compared with the gadopentetate dimeglumine set (Az value = 0.927), but the difference was not significant (p = 0.362). The sensitivity of the gadoxetic acid set (n = 51, 86.4%) was significantly higher than that of the gadopentetate dimeglumine set (n = 38, 64.4%) (p = 0.0001). Gadoxetic acid-enhanced MRI is a more sensitive diagnostic tool for detection of HCC than gadopentetate dimeglumine-enhanced MRI.

We describe a patient with Wernicke's encephalopathy who showed spontaneous upbeat nystagmus with decelerating slow phases that changed to downbeat nystagmus during upward gaze and increased during downward gaze. He also showed horizontal gaze-evoked nystagmus and impaired upward smooth pursuit. Magnetic resonance imaging demonstrated symmetric lesions involving the bilateral medial thalami, periaqueductal gray matters and inferior cerebellar peduncles. In this patient, the decelerating slow phases and disobedience to Alexander's law of upbeat nystagmus suggest both deficient (leaky) and unstable neural integrators subserving vertical eye motion. Dysfunction of the interstitial nucleus of Cajal or its descending pathway to the vestibulocerebellum via the paramedian tract cell groups may be responsible for the upbeat nystagmus and its modulation by gazes in our patient with Wernicke's encephalopathy., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Interferon (IFN) response rate in hepatitis C virus (HCV) patients has been varied with genotypes. In this study, we investigated the effects of HCV NS5A protein on IFN resistance and compared the genotypic differences of NS5A. We showed that IFN-α-, poly I:C-, and Sendai virus-induced ISRE transcriptional activities were inhibited by both genotype 1b and 2a NS5A protein. We demonstrated that not only genotype 1b but also genotype 2a NS5A exerted the similar extent of IFN-α-induced antiviral activity. We showed that NS5A derived from both genotype 1b and 2a showed no significant differential IFN responses as seen in HCV patients. These data imply that some other host factor may be involved in genotypic differences of IFN antagonism in HCV patients., (Copyright © 2010 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

Aim: To compare the diagnostic accuracy and sensitivity of gadoxetic acid-enhanced MRI and superparamagnetic iron oxide (SPIO)-enhanced magnetic resonance imaging (MRI) for the detection of hepatocellular carcinoma (HCC)., Materials and Methods: Eighty-nine patients (118 HCCs) who underwent three-dimensional gadoxetic acid-enhanced MRI and SPIO-enhanced MRI with a mean interval of 4.7 days (range 3-7 days), were included in this study. Two observers reviewed the gadoxetic acid set (unenhanced, early dynamic, 10 and 20 min hepatocyte-phase images) and SPIO set [unenhanced and ferucarbotran-enhanced T1-, T2-turbo spin-echo (TSE), and T2* weighted imaging (WI)] in consensus. Diagnostic accuracy and sensitivity were evaluated using the alternative-free response receiver operator characteristic (ROC) method., Results: The area under ROC curve (Az value) and sensitivity of the gadoxetic acid set (Az 0.964; sensitivity 90.7%) were significantly higher than those of the SPIO set (Az 0.830; sensitivity 84.7%; p<0.05). There were 14 and seven lesions that were verified only on the gadoxetic acid set and only on the SPIO set, respectively. Four HCCs were clearly revealed as hypointense only on gadoxetic acid-enhanced hepatocyte phase imaging, but were occult on other sequences, including the SPIO set., Conclusion: Gadoxetic acid-enhanced MRI is better than SPIO-enhanced MRI for the detection of HCCs.

Purpose: The aim of this study was to compare the diagnostic accuracy and sensitivity of gadoxetic acid-enhanced magnetic resonance imaging (MRI) with multi-detector row computed tomography (MDCT) for the detection of hepatocellular carcinomas (HCCs)., Materials: Sixty-two patients (81 HCCs) who underwent MDCT and gadoxetic acid-enhanced MRI using a 3-dimensional volumetric interpolated technique with a mean interval of 7 days (range, 3-11 days) were included in this study. Two observers reached a consensus on 2 sets of images: the gadoxetic acid set (unenhanced, early dynamic, 10-minute, and 20-minute hepatocyte phase images) and the 3-phase MDCT. Diagnostic accuracy and sensitivity were evaluated using the alternative-free response receiver operating characteristic method., Results: There was a trend toward increased area under the receiver operating characteristic curve (Az value) for the gadoxetic acid set (0.963) as compared with the MDCT (0.930), but no significant difference was found (P = 0.41). Sensitivity of the gadoxetic acid set (91.4%) was better than that of the MDCT (71.6%; P = 0.0001). There were 12 lesions that showed only arterial hypervascularization on MDCT but showed arterial hypervascularization and delayed hypointensity on the gadoxetic acid set., Conclusions: Gadoxetic acid-enhanced MRI, including hepatocyte phase imaging, is more sensitive than MDCT for the detection of HCCs.

Background/aims: The nonstructural 5A (NS5A) protein of hepatitis C virus (HCV) has been implicated in HCV-induced liver pathogenesis. Wnt/beta-catenin signaling has also been involved in tumorigenesis. To elucidate the molecular mechanism of HCV pathogenesis, we examined the potential effects of HCV NS5A protein on Wnt/beta-catenin signal transduction cascades., Methods: The effects of NS5A protein on beta-catenin signaling cascades in hepatic cells were investigated by luciferase reporter gene assay, confocal microscopy, immunoprecipitation assay, and immunoblot analysis., Results: beta-Catenin-mediated transcriptional activity is elevated by NS5A protein, in the context of HCV replication, and by infection of cell culture-produced HCV. NS5A protein directly interacts with endogenous beta-catenin and colocalizes with beta-catenin in the cytoplasm. NS5A protein inactivates glycogen synthase kinase 3beta and increases subsequent accumulation of beta-catenin in HepG2 cells. beta-Catenin was also accumulated in HCV patients' liver tissues. In addition, the accumulation of beta-catenin in HCV replicon cells requires both activation of phosphatidylinositol 3-kinase and inactivation of GSK3beta., Conclusions: NS5A activates beta-catenin signaling cascades through increasing the stability of beta-catenin. This modulation is accomplished by the protein interplay between viral and cellular signaling transducer. These data suggest that NS5A protein may directly be involved in Wnt/beta-catenin-mediated liver pathogenesis.

Hepatitis C virus (HCV) is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC). However, the mechanism of HCV pathogenesis is not well understood. Our previous in vitro studies suggested that non-structural 5A (NS5A) protein may play an important role in liver pathogenesis. To elucidate the mechanism of HCV-induced liver pathogenesis, we investigated the histopathological changes of liver in transgenic mice harbouring the NS5A gene. We generated transgenic mice harbouring HCV NS5A gene under the control of hepatitis B virus (HBV) enhancer. Pathological changes were analysed by immunohistochemical staining and western blot analysis. Lipid composition and reactive oxygen species (ROS) production in NS5A transgenic mice were analysed. HCV NS5A transgenic mice developed extraordinary steatosis over 6 months old and induced HCC in some mice. NS5A was co-localized with apolipoprotein A-I in fatty hepatocytes. In addition, the extraordinarily high levels of ROS, NF-kappaB and STAT3 were detected in hepatocytes of NS5A transgenic mice. These data suggest that NS5A, independent of other HCV viral proteins, may play an important role in the development of hepatic pathologies, including steatosis and hepatoceullular carcinoma in transgenic mice., (2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.)

Hepatitis delta virus (HDV) infection causes fulminant hepatitis and liver cirrhosis. To elucidate the molecular mechanism of HDV pathogenesis, we examined the effects of HDV viral proteins, the small hepatitis delta antigen (SHDAg) and the large hepatitis delta antigen (LHDAg), on NF-kappaB signaling pathway. In this study, we demonstrated that TNF-alpha-induced NF-kappaB transcriptional activation was increased by LHDAg but not by SHDAg in both HEK293 and Huh7 cells. Furthermore, LHDAg promoted TRAF2-induced NF-kappaB activation. Using coimmunoprecipitation assays, we demonstrated that both SHDAg and LHDAg interacted with TRAF2 protein. We showed that isoprenylation of LHDAg was not required for the increase of NF-kappaB activity. We further showed that only LHDAg but not SHDAg increased the TNF-alpha-mediated nuclear translocation of p65. This was accomplished by activation of IkappaBalpha degradation by LHDAg. Finally, we demonstrated that LHDAg augmented the COX-2 expression level in Huh7 cells. These data suggest that LHDAg modulates NF-kappaB signaling pathway and may contribute to HDV pathogenesis.

Hepatitis C virus (HCV) infection is often associated with hepatic steatosis and yet the molecular mechanisms of HCV-associated steatosis are poorly understood. Because sterol regulatory element-binding proteins (SREBPs) are the major transcriptional factors in lipogenic gene expression including fatty acid synthase (FAS), we examined the effects of HCV nonstructural proteins on the signaling pathways of SREBP. In this study, we demonstrated that HCV nonstructural 4B (NS4B) protein increased the transcriptional activities of SREBPs. We also showed that HCV NS4B enhanced the protein expression levels of SREBPs and FAS. This was further confirmed in the context of viral RNA replication and HCV infection. The up-regulation of both SREBP and FAS by NS4B protein required phosphatidylinositol 3-kinase activity. We also demonstrated that NS4B protein induced a lipid accumulation in hepatoma cells. In addition, NS4B protein synergistically elevated the transcriptional activity of HCV core-mediated SREBP-1. These results strongly suggest that NS4B may play an important role in HCV-associated liver pathogenesis by modulating the SREBP signaling pathway.