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Objectives: To investigate the dose length product (DLP) and outcomes of CT fluoroscopy (CTF)-guided interventions using a novel 320-detector row CT scanner with deep-learning reconstruction (DLR) and a new bow-tie filter ( i.e., Aquilion ONE Prism Edition) and compare with a 320-detector row CT system without DLR and the new bow-tie filter ( i.e., Aquilion ONE Vision Edition) (Vision)., Methods: CTF-guided interventions performed using Prism and Vision were retrospectively investigated in terms of the technical success rates, clinical success rates of biopsies, complications, DLPs of total CT scans (total DLPs) from February 2019 to January 2021. The total CT scans included pre-interventional CT scans, CTF scans during the CTF-guided procedure, additional CT scans for additional treatment, CTF scans for additional treatment, and post-interventional CT scans., Results: In this study, 87 and 85 CTF-guided interventions were performed using Vision (Vision group) and Prism (Prism group), respectively. There was no significant difference in the technical success rate (96.6% vs 98.8%, p = 0.621), clinical success rate of biopsies (92.9% vs 93.4%, p = 1.000), and minor (8.0% vs 7.1%, p = 0.807) and major (0% vs 3.5%, p = 0.119) complications between the Prism and Vision groups. The total DLPs for the Prism group were significantly lower than those for the Vision group regardless of the procedure (278 vs 548 mGy*cm, p < 0.001, in the biopsy and 246 vs 667 mGy*cm, p < 0.001, in the drainage and aspiration)., Conclusions: CTF-guided interventions on Prism reduce the total DLP without performance degradation of the intervention., Advances in Knowledge: The total DLPs of biopsies and drainages/aspirations in the Prism group decreased by 49 and 63%, respectively.

Genomic DNA replication requires replisome assembly. We show here the molecular mechanism by which CMG (GAN-MCM-GINS)-like helicase cooperates with the family D DNA polymerase (PolD) in Thermococcus kodakarensis. The archaeal GINS contains two Gins51 subunits, the C-terminal domain of which (Gins51C) interacts with GAN. We discovered that Gins51C also interacts with the N-terminal domain of PolD's DP1 subunit (DP1N) to connect two PolDs in GINS. The two replicases in the replisome should be responsible for leading- and lagging-strand synthesis, respectively. Crystal structure analysis of the DP1N-Gins51C-GAN ternary complex was provided to understand the structural basis of the connection between the helicase and DNA polymerase. Site-directed mutagenesis analysis supported the interaction mode obtained from the crystal structure. Furthermore, the assembly of helicase and replicase identified in this study is also conserved in Eukarya. PolD enhances the parental strand unwinding via stimulation of ATPase activity of the CMG-complex. This is the first evidence of the functional connection between replicase and helicase in Archaea. These results suggest that the direct interaction of PolD with CMG-helicase is critical for synchronizing strand unwinding and nascent strand synthesis and possibly provide a functional machinery for the effective progression of the replication fork., (© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.)

The eukaryotic replisome is comprised of three family-B DNA polymerases (Polα, δ and ϵ). Polα forms a stable complex with primase to synthesize short RNA-DNA primers, which are subsequently elongated by Polδ and Polϵ in concert with proliferating cell nuclear antigen (PCNA). In some species of archaea, family-D DNA polymerase (PolD) is the only DNA polymerase essential for cell viability, raising the question of how it alone conducts the bulk of DNA synthesis. We used a hyperthermophilic archaeon, Thermococcus kodakarensis, to demonstrate that PolD connects primase to the archaeal replisome before interacting with PCNA. Whereas PolD stably connects primase to GINS, a component of CMG helicase, cryo-EM analysis indicated a highly flexible PolD-primase complex. A conserved hydrophobic motif at the C-terminus of the DP2 subunit of PolD, a PIP (PCNA-Interacting Peptide) motif, was critical for the interaction with primase. The dissociation of primase was induced by DNA-dependent binding of PCNA to PolD. Point mutations in the alternative PIP-motif of DP2 abrogated the molecular switching that converts the archaeal replicase from de novo to processive synthesis mode., (© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Replication protein A (RPA) is an essential component of DNA metabolic processes. RPA binds to single-stranded DNA (ssDNA) and interacts with multiple DNA-binding proteins. In this study, we showed that two DNA polymerases, PolB and PolD, from the hyperthermophilic archaeon Thermococcus kodakarensis interact directly with RPA in vitro. RPA was expected to play a role in resolving the secondary structure, which may stop the DNA synthesis reaction, in the template ssDNA. Our in vitro DNA synthesis assay showed that the pausing was resolved by RPA for both PolB and PolD. These results supported the fact that RPA interacts with DNA polymerases as a member of the replisome and is involved in the normal progression of DNA replication forks.

The archaeal minichromosome maintenance (MCM) has DNA helicase activity, which is stimulated by GINS in several archaea. In the eukaryotic replicative helicase complex, Cdc45 forms a complex with MCM and GINS, named as CMG (Cdc45-MCM-GINS). Cdc45 shares sequence similarity with bacterial RecJ. A Cdc45/RecJ-like protein from Thermococcus kodakarensis shows a bacterial RecJ-like exonuclease activity, which is stimulated by GINS in vitro. Therefore, this archaeal Cdc45/RecJ is designated as GAN, from GINS-associated nuclease. In this study, we identified the CMG-like complex in T. kodakarensis cells. The GAN·GINS complex stimulated the MCM helicase, but MCM did not affect the nuclease activity of GAN in vitro. The gene disruption analysis showed that GAN was non-essential for its viability but the Δgan mutant did not grow at 93°C. Furthermore, the Δgan mutant showed a clear retardation in growth as compared with the parent cells under optimal conditions at 85°C. These deficiencies were recovered by introducing the gan gene encoding the nuclease deficient GAN protein back to the genome. These results suggest that the replicative helicase complex without GAN may become unstable and ineffective in replication fork progression. The nuclease activity of GAN is not related to the growth defects of the Δgan mutant cells., (© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.)

In eukaryotic DNA replication initiation, hexameric MCM (mini-chromosome maintenance) unwinds the template double-stranded DNA to form the replication fork. MCM is activated by two proteins, Cdc45 and GINS, which constitute the 'CMG' unwindosome complex together with the MCM core. The archaeal DNA replication system is quite similar to that of eukaryotes, but only limited knowledge about the DNA unwinding mechanism is available, from a structural point of view. Here, we describe the crystal structure of an archaeal GAN (GINS-associated nuclease) from Thermococcus kodakaraensis, the homolog of eukaryotic Cdc45, in both the free form and the complex with the C-terminal domain of the cognate Gins51 subunit (Gins51C). This first archaeal GAN structure exhibits a unique, 'hybrid' structure between the bacterial RecJ and the eukaryotic Cdc45. GAN possesses the conserved DHH and DHH1 domains responsible for the exonuclease activity, and an inserted CID (CMG interacting domain)-like domain structurally comparable to that in Cdc45, suggesting its dual roles as an exonuclease in DNA repair and a CMG component in DNA replication. A structural comparison of the GAN-Gins51C complex with the GINS tetramer suggests that GINS uses the mobile Gins51C as a hook to bind GAN for CMG formation., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)

The common mismatch repair system processed by MutS and MutL and their homologs was identified in Bacteria and Eukarya. However, no evidence of a functional MutS/L homolog has been reported for archaeal organisms, and it is not known whether the mismatch repair system is conserved in Archaea. Here, we describe an endonuclease that cleaves double-stranded DNA containing a mismatched base pair, from the hyperthermophilic archaeon Pyrococcus furiosus The corresponding gene revealed that the activity originates from PF0012, and we named this enzyme Endonuclease MS (EndoMS) as the mismatch-specific Endonuclease. The sequence similarity suggested that EndoMS is the ortholog of NucS isolated from Pyrococcus abyssi, published previously. Biochemical characterizations of the EndoMS homolog from Thermococcus kodakarensis clearly showed that EndoMS specifically cleaves both strands of double-stranded DNA into 5'-protruding forms, with the mismatched base pair in the central position. EndoMS cleaves G/T, G/G, T/T, T/C and A/G mismatches, with a more preference for G/T, G/G and T/T, but has very little or no effect on C/C, A/C and A/A mismatches. The discovery of this endonuclease suggests the existence of a novel mismatch repair process, initiated by the double-strand break generated by the EndoMS endonuclease, in Archaea and some Bacteria., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)

The Japanese Population-based Osteoporosis (JPOS) Cohort Study was launched in 1996 to produce a reference database of areal bone mineral density (aBMD) by dual energy X-ray absorptiometry (DXA) and bone turnover markers in the Japanese female population and to determine risk factors for osteoporotic fractures. At baseline, 3984 women aged 15 to 79 years were randomly selected to provide representative bone status data and aBMD values for the diagnosis of osteoporosis. Follow-up surveys were conducted in 1999, 2002, 2006 and 2011/12 to determine changes in aBMD and identify incident morphometry-confirmed vertebral fractures and clinical fractures. These outcomes were obtained from 2174 women who participated in at least one follow-up survey. JPOS is a unique resource of individual-level bone health information with radiological and biological archives that include DXA images, and serum, plasma and DNA for future analyses with emerging radiological and biological techniques. The JPOS dataset is not freely available, but new collaborations are encouraged. Potential collaborators are invited to contact the Secretary General (M.I.) at the administrative office of the JPOS Study Group., (© The Author 2014; all rights reserved. Published by Oxford University Press on behalf of the International Epidemiological Association.)

DNA is constantly damaged by endogenous and environmental influences. Deaminated adenine (hypoxanthine) tends to pair with cytosine and leads to the A:T→G:C transition mutation during DNA replication. Endonuclease V (EndoV) hydrolyzes the second phosphodiester bond 3' from deoxyinosine in the DNA strand, and was considered to be responsible for hypoxanthine excision repair. However, the downstream pathway after EndoV cleavage remained unclear. The activity to cleave the phosphodiester bond 5' from deoxyinosine was detected in a Pyrococcus furiosus cell extract. The protein encoded by PF1551, obtained from the mass spectrometry analysis of the purified fraction, exhibited the corresponding cleavage activity. A putative homolog from Thermococcus kodakarensis (TK0887) showed the same activity. Further biochemical analyses revealed that the purified PF1551 and TK0887 proteins recognize uracil, xanthine and the AP site, in addition to hypoxanthine. We named this endonuclease Endonuclease Q (EndoQ), as it may be involved in damaged base repair in the Thermococcals of Archaea., (© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.)

The Wnt/ß-catenin pathway is a critical stem cell regulator and plays important roles in neuroepithelial cells during early gestation. However, the role of Wnt/ß-catenin signaling in radial glia, a major neural stem cell population expanded by midgestation, remains poorly understood. This study shows that genetic ablation of ß-catenin with hGFAP-Cre mice inhibits neocortical formation by disrupting radial glial development. Reduced radial glia and intermediate progenitors are found in the ß-catenin-deficient neocortex during late gestation. Increased apoptosis and divergent localization of radial glia in the subventricular zone are also observed in the mutant neocortex. In vivo and in vitro proliferation and neurogenesis as well as oligodendrogenesis by cortical radial glia or by dissociated neural stem cells are significantly defective in the mutants. Neocortical layer patterning is not apparently altered, while astrogliogenesis is ectopically increased in the mutants. At the molecular level, the expression of the transcription factor Pax6 is dramatically diminished in the cortical radial glia and the sphere-forming neural stem cells of ß-catenin-deficient mutants. Chromatin immunoprecipitation and luciferase assays demonstrate that ß-catenin/Tcf complex binds to Pax6 promoter and induces its transcriptional activities. The forced expression of Pax6 through lentiviral transduction partially rescues the defective proliferation and neurogenesis by ß-catenin-deficient neural stem cells. Thus, Pax6 is a novel downstream target of the Wnt/ß-catenin pathway, and ß-catenin/Pax6 signaling plays critical roles in self-renewal and neurogenesis of radial glia/neural stem cells during neocortical development., (© AlphaMed Press.)

Objective: The goal of our study was to evaluate the diagnostic performance of percutaneous lung biopsy under CT-fluoroscopic guidance for ground-glass opacity (GGO) lesions., Methods: 85 percutaneous needle lung biopsies were performed in 73 patients. Specimens were obtained by core biopsy utilising an automated cutting needle and were evaluated histologically. Final diagnosis was confirmed by independent surgical pathology, independent culture results or clinical follow-up., Results: Rates of adequate specimens obtained and of precise diagnosis by needle biopsy were 92.9% (79/85) and 90.6% (77/85) of evaluated lung lesions, respectively. Precise diagnosis was achieved in 87.1% (27/31) of lesions ≤10 mm in diameter, 90.0% (36/40) of lesions >10 mm to ≤20 mm and 100.0% (14/14) of lesions >20 mm. Precision in diagnosing GGO lesions according to the GGO component was 73.9% (17/23) for pure GGO lesions and 96.8% (60/62) for part-solid GGO lesions. Obtaining a precise diagnosis did not differ significantly according to the lesion size (p=0.3840), but differences were significant according to the GGO component (p=0.0047). Malignancy was accurately diagnosed in 35 of 36 malignant lesions for which surgery was later performed. The specific cell type determined from specimens obtained by needle biopsy was exactly the same as the final histological diagnosis obtained after surgery in 20 lesions., Conclusion: Tissue-core lung biopsy under CT-fluoroscopic guidance for a GGO lesion provides a high degree of diagnostic accuracy but is less reliable for determining the specific cell type., Advances in Knowledge: Percutaneous lung biopsy under CT-fluoroscopic guidance for GGO is useful in differentiating malignancy.

Objective: The aim of this study was to compare two different automated biopsy needles, a fully automated biopsy needle (Monopty; Bard, Covington, GA) and a semi-automated biopsy needle (Temno; Bauer Medical, Clearwater, FL), for lung biopsy., Methods: 50 consecutive percutaneous lung biopsies using the Monopty needle between June 2006 and January 2007 and 66 consecutive lung biopsies for 1 nodule in each session using the Temno needle between February 2007 and August 2008 were performed under CT fluoroscopic guidance followed by histopathological evaluation., Results: In 42/50 lung biopsies performed with the Monopty needle and 54/66 lung biopsies performed with the Temno needle, the final diagnosis was confirmed by independent surgical pathological findings or clinical follow-up. Sufficient samples for histopathological evaluation were obtained in all 50 (100%) biopsies using the Monopty needle and in 55 (83.3%) of the 66 biopsies using the Temno needle (p<0.01). Accurate diagnosis was achieved in 41 (97.6%) of 42 biopsies using the Monopty needle and in 45 (83.3%) of 54 biopsies using the Temno needle (p=0.04). Biopsy-induced complications were pneumothorax, haemoptysis and haemothorax in 44.0%, 10.0% and 6.0% of biopsies, respectively, using the Monopty needle and in 48.3%, 8.3% and 3.3%, respectively, using the Temno needle., Conclusion: There is a possibility that a fully automated biopsy needle such as the Monopty is more useful for CT scan-guided lung biopsy than semi-automated biopsy needles.

OsGEN-L has a 5'-flap endonuclease activity and plays an essential role in rice microspore development. The Class 4 RAD2/XPG family nucleases, including OsGEN-L, were recently found to have resolving activity for the Holliday junction (HJ), the intermediate of DNA strand recombination. In this study, we performed a detailed characterization of OsGEN-L, as a structure-specific endonuclease. Highly purified OsGEN-L was prepared as the full-length protein for in vitro endonuclease assays using various structured DNAs, and the 5'-flap endonuclease activity, which is stimulated in a PCNA-dependent manner, was demonstrated. In addition, the in vitro HJ resolving activity of OsGEN-L represents the first such activity originating from plant cells. OsGEN-L cleaved HJ at symmetrically related sites of the branch point. However, the two branched strands seemed to be cleaved individually, and not cooperatively, by each OsGEN-L monomer protein. The substrate specificity suggests that OsGEN-L functions in multiple processes of DNA metabolism in rice cells.

Objective: This study evaluated the feasibility and safety of the transjugular intrahepatic portosystemic shunt (TIPS) procedure using the hepatic artery-targeting guidewire technique for the puncture step., Methods: We retrospectively reviewed 11 consecutive patients (5 men and 6 women, aged 46-76 years (mean 64 years)) with portal hypertension in whom the TIPS procedure was performed. As the first step in the TIPS procedure in all cases, a micro-guidewire was inserted into the hepatic arterial branch accompanying the portal venous branch through a microcatheter coaxially advanced from a 5-French catheter positioned in the coeliac or common hepatic artery. At the puncture step, the tip of the metallic cannula was aimed 1 cm posterior to the distal part of this micro-guidewire, after which the TIPS procedure was performed. Success rate, number of punctures and complications were evaluated., Results: The TIPS procedure was successfully performed in all 11 patients. The mean number of punctures until success in entering the targeted portal venous branch was 5 (range 1-14). In 3 patients (27%), the right portal venous branch was entered at the first puncture attempt. The hepatic artery was punctured once in one patient and the bile duct was punctured once in another patient. No serious procedure-induced complications occurred., Conclusion: The TIPS procedure can be accomplished safely, precisely and relatively easily using the hepatic artery-targeting guidewire technique.

We used a retrospective non-randomised study to investigate the clinical effect of selective embolisation of the right gastric artery before hepatic arterial infusion chemotherapy (HAIC) using a port-catheter system. We evaluated whether the hepatic artery or the left gastric artery is the better approach for selecting the right gastric artery. A total of 367 patients (244 men and 123 women; mean age, 64.1 years) with unresectable advanced liver cancer underwent percutaneous implantation of a port-catheter system. In 294 of these patients, right gastric arterial embolisation with microcoils was attempted before placement of the port-catheter system to prevent gastric mucosal lesions. Approach was either through the hepatic artery (175 patients) or through the left gastric artery (119 patients), with success rates in catheterising the right gastric artery of 78.3% and 77.3%, respectively. If the attempt was unsuccessful, the catheter was redirected to the alternative approach, which increased the final success rate to 96.3%. Only seven patients experienced gastroduodenal mucosal lesions acutely after HAIC, as revealed by endoscopy. Embolisation of the right gastric artery is a feasible procedure that can reduce the incidence of gastric mucosal lesions associated with HAIC. Approach through either the hepatic artery or the left gastric artery is equally acceptable.

Objectives: To determine whether aquaporins (AQPs) are expressed in the synovial tissues of patients with OA and RA, and to examine the patterns of expression in patients with and without hydrarthrosis., Methods: AQPs were detected in synovial tissue samples from patients with OA and RA using RT-PCR and immunohistochemistry. Fibroblast-like synoviocytes (FLSs) from patients with OA and RA were cultured and stimulated with TNF-alpha. The expression of AQPs in FLSs was examined using RT-PCR and western blot analyses and the function of aquaglyceroporins was examined by a glycerol uptake assay., Results: AQP1, -3 and -9 mRNAs were expressed in synovial tissues from patients with OA and RA. AQP1, -3 and -9 proteins were also detected by immunohistochemistry. AQP9 mRNA was expressed more strongly in the synovial tissues of OA patients with hydrarthrosis than those without. AQP9 mRNA and protein expression were strongly induced with TNF-alpha treatment in FLSs, whereas the expression of AQP1 and -3 mRNAs was not induced with TNF-alpha treatment. AQP9 as an aquaglyceroporin was induced by TNF-alpha., Conclusions: AQP9 mRNA was detected in synovial tissues from OA and RA patients with hydrarthrosis. AQP9 expression was strongly induced in FLSs with TNF-alpha. Although the functions of AQP1, -3 and -9 in synovial tissues remain to be elucidated, it suggested that AQP9 might be related to the pathogenesis of hydrarthrosis and inflammatory synovitis.

The purpose of this study was to assess the effectiveness of lymphangiography as a treatment for various chyle leakages. Pedal lymphangiography was performed in 9 patients (6 men and 3 women; mean age, 59 years) who were unlikely to be cured only by conservative treatment - a low-fat medium-chain triglyceride diet, total parenteral nutrition and insertion of a drainage tube - and in whom chylothorax (n = 5), chylous ascites (n = 2) and lymphatic fistulae (n = 2) were refractory to conservative treatment. In 7 of these 9 patients (78%), we could detect the chyle leakage sites. In 8 of the 9 patients (89%), lymphatic leakage was stopped after lymphangiography, and surgical re-intervention was avoided. No cases had a recurrence of chyle leakage during follow-up (range, 1-54 months). Lymphangiography is effective not only for diagnosis but also as treatment for various chyle leakages. Early lymphangiography is therefore recommended for patients with chyle leakages who are unlikely to be cured by conservative treatment only.

We encountered a patient with a gastric varix that drained through the left inferior phrenic vein, which directly entered the inferior vena cava at the point just inferior to the diaphragm. In this patient, gastrorenal shunt was not seen. Balloon-occluded retrograde transvenous obliteration of the gastric varix was performed, in which 50% glucose and 5% ethanolamine oleate-iopamidol were injected as sclerosing agents while the balloon was inflated in the left inferior phrenic vein. 1 week after the procedure, the disappearance of enhancement in the gastric varix was confirmed on contrast-enhanced multidetector row CT. Furthermore, a significant reduction in the size of the varix was confirmed on endoscopic examination 4 months later.

The aim of this study is to evaluate the feasibility and safety of CT fluoroscopic-guided needle biopsy with the use of the "I-I device", which was developed to assist in precisely advancing the needle while avoiding irradiation to the operator's hand. Using the "I-I device" under CT fluoroscopic guidance, 131 percutaneous needle lung biopsies were performed followed by histopathological evaluation. The final diagnosis was confirmed by independent surgical pathological findings or clinical follow-up. The rate of success in obtaining specimens adequate for histopathological analysis was 100% (131/131). For the 104 lesions that we were able to follow up, sensitivity, specificity and accuracy in diagnosing malignancy were 93.8%, 100% and 95.2%, respectively. In 51 lesions for which surgery was performed, the specific cell type was characterized in 98.0% (50/51; malignant, n = 38; benign, n = 12). The specific cell type was precisely diagnosed and confirmed after surgery in 36 malignant lesions and 8 benign lesions. Biopsy-induced complications were pneumothorax in 34.0% (44/131) and haemoptysis in 9.9% (13/131). None of the patients had serious complications. In conclusion, CT fluoroscopic-guided lung biopsy with use of the "I-I device" provides a high degree of diagnostic accuracy, allows specific characterization of lung nodules and can be performed safely.

We report a successful percutaneous transluminal angioplasty using a pull-through technique for the treatment of a hepatic arterial occlusion caused by iatrogenic subintimal dissection during the percutaneous placement of a port-catheter system.

Pseudoaneurysms can arise due to localized arterial wall disruption, owing to various factors such as inflammation, trauma, neoplasm, or surgical procedures. Once ruptured, bleeding can be life-threatening. Thus, a treatment for pseudoaneurysm is necessary. We describe a case of post-surgical ruptured pseudoaneurysm of the lingual artery that was successfully treated by transcatheter embolisation with n-butyl cyanoacrylate (NCBA).

Measurements of cosmic-ray neutron dose rates with a balloon in Sanriku, Japan (geographic location: 39 degrees N, 142 degrees E; corresponding geomagnetic latitude: 30 degrees N) were conducted at an altitude from 0.2 to 25 km on 25-26 August 2004 when solar activity was at an average level. Neutron dose rates given as ambient dose equivalent rates (H(10)) were measured with high-sensitive neutron dose equivalent counters and electronic silicon personal dosimeters (EPDs). The neutron dose rates increased with increasing altitude, but they were saturated around 15-20 km and decreased with increasing altitude beyond 20 km. The neutron ambient dose equivalent rate was 1.5 microSv/h(- 1) at 20 km. Measured values were corrected for the deviation of the energy response of the dose equivalent counter from the fluence-to-ambient dose equivalent conversion coefficient, and the corrected values were very close to the calculated values with EPCARD. On the other hand, neutron measurements by the EPDs gave about 10 times overestimation because of the high sensitivity to cosmic-ray protons.

We report a 60-year-old man who after undergoing surgical repair of an abdominal aortic aneurysm with a bifurcated graft subsequently developed a proximal anastomotic pseudoaneurysm, which was successfully treated by embolisation with n-butyl cyanoacrylate.

Interventional radiological treatment by transcatheter embolisation has been used to treat patients with symptomatic intrahepatic portosystemic venous shunt (IPSVS). We present a case of symptomatic IPSVS treated by both retrograde and antegrade transcatheter embolisation using coils and n-butyl cyanoacrylate.

The purpose of this study was to evaluate the filtering effect of the Gunther tulip retrievable vena cava filter (GTF) during treatment of deep venous thrombosis (DVT) in the lower extremity using various interventional radiological procedures. Subjects of the study were all 17 consecutive patients (8 women, 9 men; age range 18-87 years; mean age 55.9 years) with symptomatic lower limb DVT referred for interventional radiological treatment between February 2001 and September 2004. In all of these patients, the GTF was implanted during interventional radiological treatment. Trapped thrombus in the filter was evaluated with venocavography performed repeatedly during the treatment for DVT. Implantation of a total of 29 GTFs was successfully performed in the 17 patients. In 10 (58.8%), more than 2 filters were subsequently implanted to prolong implantation time. Also in 10 patients, the DVT resolved after therapy and retrieval of the final GTF was successful with one exception. Worsening of or new formation of pulmonary embolism was avoided in all patients. In 8 (47.1%) of the 17 patients, a trapped thrombus in the GTF was observed during treatment for DVT. In six patients the trapped thrombus was large, filling more than half the height of the filter. In conclusion, we found that the GTF is effective in filtering the relieved thrombus from DVT in the lower extremity and in protecting against movement of the thrombus to the pulmonary artery during therapies with interventional radiological procedures.

We report a 34-year-old man with the complication of chylous ascites after retroperitoneal lymphadenectomy that was refractory to various conservative therapies. Because surgical treatment for chylous ascites was considered, lymphangiography was performed to identify the area of leakage of chyle, after which the chylous ascites spontaneously healed.

We report a case of an intrahepatic portosystemic venous shunt (IPSVS) passing through the left inferior phrenic vein and draining into the left renal vein and then to the inferior vena cava. We incidentally detected the IPSVS while performing a transcatheter arterial embolisation for the treatment of hepatocellular carcinoma. IPSVS with the left inferior phrenic vein as the draining vein is very rare. The complete pathway of this IPSVS could be clearly demonstrated by multidetector row CT during arterial portography.

This report describes a 58-year-old female with adenosquamous carcinoma (ASC) of the liver presenting with right lower abdominal pain. In most ASC of the liver, surgery is the first choice of treatment. However, surgery often seems to be ineffective because of the aggressive behaviour of this disease. At surgical laparotomy in the present case, there was a large tumour occupying the entire right lobe of the liver and invading the diaphragm. Thus, partial hepatectomy was performed. For the residual tumour, we performed repeated transcatheter arterial infusion chemotherapy with degradable starch microspheres (DSM). After four sessions of transcatheter arterial infusion chemotherapy, tumour size decreased remarkably and tumour markers had also decreased. Despite the poor prognosis, the patient remains alive and well 12 months after laparotomy. It is suggested that minimally invasive transcatheter infusion chemotherapy with DSM can be an effective treatment preserving a high quality of life.

We report a case of small pulmonary arteriovenous fistulae precisely diagnosed by a combination of selective pulmonary arteriography and scintigraphy during (99)Tc(m)-macroaggregated albumin injection via the peripheral pulmonary artery.

A cDNA encoding rye seed chitinase-a (RSC-a) was cloned by rapid amplification of cDNA ends and PCR procedures. It consists of 1,191 nucleotides and encodes an open reading frame of 321 amino acid residues. Recombinant RSC-a (rRSC-a) was produced in the oxidative cytoplasm of Escherichia coli Origami(DE3) in a soluble form by inducing bacteria at a low temperature (20 degrees C). Purified rRSC-a showed properties similar to the original enzyme from rye seeds in terms of chitinase activity toward a soluble substrate, glycolchitin, and an insoluble substrate, chitin beads, in chitin-binding ability to chitin, and in antifungal activity against Trichoderma sp. in vitro. rRSC-a mutants were subsequently produced and purified by the same procedures as those for rRSC-a. Mutation of Trp23 to Ala decreased the chitinase activity toward both substrates and impaired the chitin-binding ability. Furthermore, the antifungal activity of this mutant was weakened with increasing of the NaCl concentration in the culture medium. Complete abolishment of both activities was observed upon the mutation of Glu126 to Gln. The roles of these residues in both activities are discussed.

The purpose of our study is to evaluate the safety of insertion and removal of the retrievable vena cava filter and the efficacy of this filter in preventing pulmonary embolism during treatment of stenosis or obstruction of the lower extremity due to deep venous thrombosis. We retrospectively evaluated placement of 10 filters in 6 patients with lower extremity deep venous thrombosis who had undergone various treatments, including those utilizing techniques of interventional radiology. Worsening of or the new formation of pulmonary embolism was prevented in all patients. All attempts at both implantation and retrieval of the filter were safely accomplished. No complication related to filter placement occurred. Filters were left in the vena cava for permanent use when deep venous thrombosis was refractory to treatment. These results demonstrate that both placement and removal of the retrievable vena cava filter can be accomplished safely and that this type of filter is useful to prevent pulmonary embolism during treatment of deep venous thrombosis of the lower extremity. Further, the use of a retrievable vena cava filter might be preferred over that of a temporary vena cava filter, especially if further implantation of a filter becomes necessary since replacement with a permanent filter would not be required when using a retrievable vena cava filter.

We expressed chitinase-1 (TBC-1) from tulip bulbs (Tulipa bakeri) in E. coli cells and used site-directed mutagenesis to identify amino acid residues essential for catalytic activity. Mutations at Glu-125 and Trp-251 completely abolished enzyme activity, and activity decreased with mutations at Asp-123 and Trp-172 when glycolchitin was the substrate. Activity changed with the mutations of Trp-251 to one of several amino acids with side-chains of little hydrophobicity, suggesting that hydrophobic interaction of Trp-251 is important for the activity. Molecular dynamics (MD) simulation analysis with hevamine as the model compound showed that the distance between Asp-123 and Glu-125 was extended by mutation of Trp-251. Kinetic studies of Trp-251-mutated chitinases confirmed these various phenomena. The results suggested that Glu-125 and Trp-251 are essential for enzyme activity and that Trp-251 had a direct role in ligand binding.

The aim of this study was to evaluate the incidence of brain infarction after percutaneous implantation of a port-catheter system via the left subclavian artery for hepatic arterial infusion chemotherapy. In 90 patients with inoperable liver cancer, a port-catheter system was implanted via the left subclavian artery. In 5 patients (5.6%) brain infarction occurred after port-catheter implantation. In one patient (1.1%) thrombi formed around the catheter, as confirmed by autopsy. The risk of brain infarction should be taken into consideration when a trans-left subclavian arterial access route is used for the percutaneous implantation of a port-catheter system for hepatic arterial infusion chemotherapy.

The antifungal activities of rye seed chitinase-a (RSC-a, class I) and -c (RSC-c, class II) were studied in detail using two different bioassays with Trichoderma sp. as well as binding and degradation experiments with the cell walls prepared from its mycelia. RSC-a inhibited more strongly the re-extension of the hyphae, containing mainly mature cells, than RSC-c did. Upon incubation of the fungus with fluorescent chitinases, FITC-labeled RSC-a was found to be located in the hyphal tips, lateral walls, and septa, while FITC-labeled RSC-c was only in the hyphal tip. RSC-a had a greater affinity for the cell walls than RSC-c. RSC-a liberated a larger amount of reducing sugar from the cell walls than RSC-c did. These results inferred that RSC-a first binds to the lateral walls and septa, consisting of the mature cell walls, and degrades mature chitin fiber, while RSC-c binds only to the hyphal tip followed by degradation of only nascent chitin. As a result, RSC-a inhibited fungal growth more effectively than RSC-c. Furthermore, it was suggested that the chitin-binding domain in RSC-a assists the antifungal action of RSC-a by binding to the fungal hypha.

We cloned a complete cDNA encoding rye seed chitinase-c, designated RSC-c, by rapid amplification of cDNA end and PCR procedures. The cDNA of RSC-c consists of 1,018 nucleotides and includes an open reading frame encoding a polypeptide of 266 amino acid residues. A recombinant RSC-c was produced by expression in Escherichia coli Origami(DE3) and purified. rRSC-c had almost the same chitinase activity toward glycolchitin and antifungal activity against Trichoderma sp. as the authentic RSC-c did. RSC-c mutants were subsequently constructed and characterized with respect to their chitinase and antifungal activities. Mutation of Glu67 to Gln completely abolished the chitinase activity and diminished the antifungal activity. Considerable decreases in both activities were observed in the mutations of Trp72 and Ser120 to Ala, and Glu89 to Gln. The roles of these residues in the catalytic event of RSC-c are discussed.

In order to understand a physiological role of chitinases in rye, the localization and accumulation of rye seed chitinase-a and -c (RSC-a and -c) in the seeds were studied by immunochemical methods. An antiserum specific to the chitin-binding domain (CB-domain), which is an N-terminal part of RSC-a, and an antiserum specific to the catalytic region of RSC-a and RSC-c were used. An immunoblot analysis detected both RSC-a and RSC-c in the endosperm of the rye seed. Immunohistochemical staining indicated that RSC-a was localized in only the aleurone cells, whereas RSC-c existed at least in the starchy endosperm and was also likely to exist in the aleurone cells. It was found by ELISA and an immunoblot analysis that RSC-a and -c accumulated in the seed during the later stage of development. Both chitinases and the Cat-domain exhibited antifungal activity toward Trichoderma species, while the CB-domain did not. Observation of the inhibition of hyphal growth of the T. species suggests that the two chitinases acted in different ways.

The haemodynamics in non-tumorous abnormalities on CT arterial portography (CTAP) owing to cholecystic venous direct inflow to the liver were compared with the haemodynamics in hepatocellular carcinoma. 53 patients who simultaneously underwent CTAP and CT during hepatic arteriography (CTHA) to detect hepatocellular carcinoma had the late phase added to CTHA. Changes in size, shape and pattern of 47 non-tumorous enhancement abnormalities on the liver around the gall bladder or in the dorsum of segment IV between the early and late phases on biphasic CTHA as well as of 60 tumorous lesions were determined. Enhancement on biphasic CTHA was seen in all 47 lesions with a non-tumorous portal defect (early phase alone, n=8; late phase alone, n = 3; both, n = 36). In these 47 lesions, the size and the shape of enhancement changed in 63.8% and 51.1%, respectively, between the early and late phases on CTHA; the pattern of enhancement did not change in 72.3%. On the other hand, the size of enhancement on biphasic CTHA changed in only 16.7% of 60 tumours, and the shape in only 5%, although the enhancement pattern changed in a large proportion (80%). In conclusion, owing to the difference in haemodynamics, non-tumorous abnormalities caused by cholecystic venous inflow and tumours are clearly delineated on biphasic CTHA. Thus, adding the late phase to previous single phase CTHA (i.e. performing biphasic CTHA) is useful in differentiating the two entities.

This study was conducted to investigate the haemodynamics of the liver parenchyma in the presence of intrahepatic portosystemic venous shunt. 3 patients with intrahepatic portosystemic venous shunts and 24 patients with normal intrahepatic haemodynamics underwent both CT arterial portography and CT during hepatic arteriography. Angiographic findings with helical CT were compared, and CT attenuated values were measured in both groups. The liver parenchyma on CT arterial portography had lower attenuation than on CT during hepatic arteriography in all patients with intrahepatic portosystemic venous shunts. Overall average CT attenuation was 92.2 +/- 7.7 Hounsfield units (HU) on CT arterial portography and 149.9 +/- 8.5 HU after CT during hepatic arteriography, with the opposite findings in all patients without intrahepatic portosystemic venous shunt: CT attenuation 142.0 +/- 25.7 HU on CT arterial portography and 100.7 +/- 16.4 HU after CT during hepatic arteriography. In conclusion, the portal venous supply to the liver parenchyma decreased due to intrahepatic portosystemic venous shunts, with a compensatory increase in hepatic arterial blood supply.

A cDNA encoding tulip bulb chitinase-1 (TBC-1) was cloned using a combination of immunoscreening from a lambda ZAP cDNA library with anti-TBC-1 antiserum and the 5' rapid amplification of cDNA end (RACE) method, and sequenced. The cDNA consists of 1,106 nucleotides and included an open reading frame encoding a polypeptide of 314 amino acids. Comparison of the deduced amino acid sequence and the determined protein sequence indicated the presence of a signal peptide and an extra peptide composed of 26 and 13 amino acids at the N- and C-termini, respectively. The deduced sequence of TBC-1 had 10-20% and 63% sequence similarities to plant class III chitinases and gladiolus bulb class IIIb chitinase (GBC-a), respectively. The cDNA encoding mature TBC-1 was amplified by polymerase chain reaction (PCR), ligated into the expression vector pET-22b, and expressed in Escherichia coli BL21(DE3). The recombinant TBC-1 (rTBC-1) expressed in E. coli was purified by gel filtration followed by ion-exchange chromatography. Specific activity of the rTBC-1 was almost same as the authentic TBC-1 toward glycolchitin. This is the first report on the cDNA cloning of a class III chitinase having C-terminal extra peptide.

The causes of non-tumorous abnormalities in the hepatic hilum seen on CT hepatic arteriography were investigated. 13 patients with non-tumorous defects of portal perfusion in the hepatic hilum on CT arterial portography underwent both CT hepatic arteriography from the common hepatic artery and CT obtained during proper hepatic arteriography. The findings of non-tumorous portal defects on these two angiographic studies using helical CT were compared. In the 13 patients, 14 non-tumorous defects of portal perfusion in the hepatic hilum on CT arterial portography were detected as enhanced areas in 10 regions (dorsum of segment IV, 7/10; dorsum of the lateral segment, 3/4) on CT hepatic arteriography via the common hepatic artery, but none were enhanced on CT obtained during proper hepatic arteriography. In conclusion, the main cause of non-tumorous enhancement in the hepatic hilum seen on CT hepatic arteriography is non-portal direct inflow via the parabiliary venous system.

The positions of disulfide bonds of rye seed chitinase-a (RSC-a) were identified by the isolation of disulfide-containing peptides produced with enzymatic and/or chemical cleavages of RSC-a, followed by sequencing them. An unequivocal assignment of disulfide bonds in this enzyme was as follows: Cys3-Cysl8, Cys12-Cys24, Cys15-Cys42, Cys17-Cys31, and Cys35-Cys39 in the chitin-binding domain (CB domain), Cys82-Cys144, Cys156-Cys164, and Cys282-Cys295 in the catalytic domain (Cat domain), and Cys263 was a free form.

Direct inflow of the aberrant right gastric vein independent of the portal venous system to the liver parenchyma is a well known cause of non-tumorous abnormalities on CT angiography. However, there have been no previous reports of aberrant gastric venous drainage as the cause of non-tumorous abnormalities in the left lobe of the liver parenchyma adjacent to the falciform ligament on CT angiography. We present a case of direct inflow of an aberrant gastric vein to this region of the liver, which influenced the non-tumorous enhancement shown by CT hepatic arteriography.

Intrahepatic portosystemic venous shunt (IPSVS) is relatively uncommon, and is usually associated with chronic hepatitis or cirrhosis. We present a case of IPSVS that was considered to be caused by increased blood flow from a large abdominal tumour. The characteristic intrahepatic haemodynamics were demonstrated by CT angiography.

The correlation between an aberrant pancreaticoduodenal vein and a portal perfusion defect in the dorsal part of segment IV as demonstrated on CT arterial portography (CTAP) was investigated. 14 patients with non-tumorous defects of portal perfusion in the dorsal part of segment IV of the liver parenchyma, shown on CTAP underwent CT during pancreaticoduodenal arteriography. The defect on CTAP was shown as an enhanced area resulting from non-portal venous inflow in eight (57%) of 14 patients on CT during pancreaticoduodenal arteriography. In conclusion, the non-portal venous supply via an aberrant pancreaticoduodenal vein occasionally causes a defect of portal perfusion in the dorsal part of segment IV on CT arterial portography.

In some cases, focal fatty change of the liver parenchyma has been attributed to local systemic venous inflow replacing the portal venous supply. However, no reports of focal fatty change in the dorsal portion of segment IV of the liver caused by direct inflow of an aberrant pancreaticoduodenal vein (APDV) through the parabiliary venous system (PVS) are available. We report a 60-year-old woman with focal fatty change in segment IV of the liver which occurred after gastrectomy with APDV shown on CT performed during selective gastroduodenal arteriography.

The complete amino acid sequences of tulip bulb chitinase-1 and -2 (TBC-1 and -2) were determined. The sequences of the TBC-1 and TBC-2 were solved by analysis of peptides derived by enzymatic digestions as well as by chemical cleavages with cyanogen bromide (CNBr), o-iodosobenzoic acid, and hydroxylamine. TBC-1 and TBC-2 both consisted of 275 amino acid residues and had molecular masses of 30,825 and 30,863, respectively. They shared 247 identical residues, that is 90% identity. Comparison of their sequences with that of gladiolus bulb class IIIb chitinase-a (GBC-a) showed that 63% of the residues of both TBC-1 and TBC-2 are identical to that of GBC-a. From these results, it was seen that TBC-1 and -2 are class IIIb chitinases. The characteristic difference in specific activity between TBC-1 and -2 was also discussed on the basis of their amino acid sequences.

The complete amino acid sequence of pokeweed leaf chitinase-A was determined. First all 11 tryptic peptides from the reduced and S-carboxymethylated form of the enzyme were sequenced. Then the same form of the enzyme was cleaved with cyanogen bromide, giving three fragments. The fragments were digested with chymotrypsin or Staphylococcus aureus V8 protease. Last, the 11 tryptic peptides were put in order. Of seven cysteine residues, six were linked by disulfide bonds (between Cys25 and Cys74, Cys89 and Cys98, and Cys195 and Cys208); Cys176 was free. The enzyme consisted of 208 amino acid residues and had a molecular weight of 22,391. It consisted of only one polypeptide chain without a chitin-binding domain. The length of the chain was almost the same as that of the catalytic domains of class IL chitinases. These findings suggested that this enzyme is a new kind of class IIL chitinase, although its sequence resembles that of catalytic domains of class IL chitinases more than that of the class IIL chitinases reported so far. Discussion on the involvement of specific tryptophan residue in the active site of PLC-A is also given based on the sequence similarity with rye seed chitinase-c.

Six chitinase isoforms, designated TBC-1 to TBC-6, were purified to homogenity from the bulbs of four species (Tulipa bakeri, T. tarda, T. turkestanica, and T. praestans) of the genus Tulipa by CM-cellulose column chromatography, Butyl-Toyopearl 650M hydrophobic column chromatography, gel filtration on Sephadex G-75, and Mono-S fast protein liquid chromatography (FPLC). The chitinases had molecular weights of 30,000 and isoelectric points of 5.2 to 6.1. These chitinases were found to proteins with similar amino acid compositions and N-terminal sequences. The tulip chitinases all had two half-cystine residues, one more than gladiolus bulb class IIIb chitinase, but many fewer than chitinases of plant class I (15-17 Cys residues/mol), II (5-8 Cys residues/mol), or III (6 Cys residues/mol). The N-terminal sequences of tulip chitinases were similar to the sequence of the gladiolus chitinase, but did not resemble sequence of any class of plant chitinase. The optimal pH of these chitinases toward glycolchitin was pH 5. TBC-1 hydrolyzed (GlcNAc)6 into (GlcNAc)2, (GlcNAc)3, and (GlcNAc)4, and hydrolyzed (GlcNAc)5 into (GlcNAc)2 and (GlcNAc)3.

Carboxyl groups of rye seed chitinase-c (RSC-c) were modified with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and glycine ethyl ester (GEE) at pH 5.5 and 5 degrees C in the presence and absence of (GlcNAc)4. In the absence of (GlcNAc)4, 5.2 carboxyl groups were modified by 90 min-reaction and the chitinase activity was reduced to 2.0%, while in the presence of (GlcNAc)4, 4.6 carboxyl groups were modified and 72% of the activity was retained. To identify the carboxyl group protected by (GlcNAc)4 from the modification, RSC-c was first modified with EDC and GEE in the presence of (GlcNAc)4 and then radiolabeled with EDC and [14C]GEE in the absence of (GlcNAc)4. Analyses of the radioactive peptides from the tryptic and chymotryptic digests of radiolabeled RSC-c showed that the main radiolabeled carboxyl group is that of Asp95, suggesting that Asp95 is located at or near substrate-binding site of RSC-c.