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Objectives: We evaluated the thrombus-vaporizing effect of excimer laser coronary angioplasty (ELCA) in patients with ST-segment elevation myocardial infarction (STEMI) by optical coherence tomography (OCT)., Background: Larger intracoronary thrombus elevates the risk of interventional treatment and mortality in patients with STEMI., Methods: A total of 92 patients with STEMI who presented within 24 hours from the onset and underwent ELCA following manual aspiration thrombectomy (MT) were analyzed., Results: The mean baseline thrombolysis in myocardial infarction flow grade was 0.4 ± 0.6, which subsequently improved to 2.3 ± 0.7 after MT ( p < 0.0001) and 2.7 ± 0.5 after ELCA ( p =0.0001). The median residual thrombus volume after MT was 65.7 mm 3 , which significantly reduced to 47.5 mm 3 after ELCA ( p < 0.0001). Plaque rupture was identified by OCT in only 22 cases (23.9%) after MT, but was distinguishable in 36 additional cases after ELCA (total: 58 cases; 63.0%). Ruptured lesions contained a higher proportion of red thrombus than nonruptured lesions (75.9% vs. 43.3%, p =0.001). Significantly larger thrombus burden after MT (69.6 mm 3 vs. 56.3 mm 3 , p < 0.05) and greater thrombus reduction by ELCA (21.2 mm 3 vs. 11.8 mm 3 , p < 0.01) were observed in ruptured lesions than nonruptured lesions., Conclusions: ELCA effectively vaporized intracoronary thrombus in patients with STEMI even after MT. Lesions with plaque rupture contained larger thrombus burden that was frequently characterized by red thrombus and more effectively reduced by ELCA., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this article., (Copyright © 2021 Yuki Yamanaka et al.)

Purpose: To evaluate the cost-effectiveness of percutaneous cryoablation (PCA) versus robot-assisted partial nephrectomy (RPN) in patients with small renal tumors (T1a stage), considering perioperative complications., Materials and Methods: Retrospective study from November 2008 to April 2017 of 122 patients with a T1a renal mass who after being analyzed by a multidisciplinary board underwent to PCA (59 patients) or RPN (63 patients). Hospital costs in US dollars, and clinical and tumor data were compared. Non-complicated intervention was considered as an effective outcome. A hypothetical model of possible complications based on Clavien-Dindo classification (CDC) was built, grouping them into mild (CDC I and II) and severe (CDC III and IV). A decision tree model was structured from complications of published data., Results: Patients who underwent PCA were older (62.5 vs. 52.8 years old, p < 0.001), presented with more coronary disease and previous renal cancer (25.4% vs. 10.1%, p = 0.023 and 38% vs. 7.2%,  p < 0.001, respectively). Patients treated with PCA had a higher preoperative risk (American Society of Anesthesiologists-ASA ≥ 3) than those in the RPN group (25.4% vs. 0%, p < 0.001). Average operative time was significantly lower with PCA than RPN (99.92 ± 29.05 min vs. 129.28 ± 54.85 min, p < 0.001). Average hospitalization time for PCA was 2.2 ± 2.95 days, significantly lower than RPN (mean 3.03 ± 1.49 days, p = 0.04). The average total cost of PCA was significantly lower than RPN (US$12,435 ± 6,176 vs. US$19,399 ± 6,047, p < 0.001). The incremental effectiveness was 5% higher comparing PCA with RPN, resulting a cost-saving result in favor of PCA., Conclusion: PCA was the dominant strategy (less costly and more effective) compared to RPN, considering occurrence of perioperative complications.

The C-H activation/annulation reaction of various benzamides with fluoroalkylated alkynes in the presence of a Co(acac) 2 ·2H 2 O catalyst proceeded very smoothly to give the corresponding 3- and 4-fluoroalkylated isoquinolinones in excellent yields with approximately 70% regioselectivities. These regioisomers could be successfully separated and obtained in pure form. Major or minor regioisomers were determined as 4- or 3-fluoroalkylated isoquinolinones, respectively, based on X-ray crystallographic analyses.

Regioselective cobalt-catalyzed [2+2+2] cycloaddition using fluorine-containing diynes with nitriles was described. Cycloaddition of fluorinated diynes with nitriles under the influence of CoCl 2 (phen), zinc bromide, and zinc dust in dichloroethane at 80°C for 3 h took place smoothly, exclusively affording the corresponding α -fluoroalkylated pyridines in excellent yields. In addition, dinitriles as substrate were also found to be suitable for this reaction, giving the corresponding fluoroalkylated bipyridine derivatives in excellent yields.

Objective: We report the usefulness of the adjunctive technique using a 3.4-Fr TACTICS catheter, which is a distal access catheter (DAC) for coil embolization., Methods: Patients who underwent coil embolization with an adjunctive technique using a TACTICS catheter between October 2018 and May 2019 were retrospectively analyzed., Results: In all, 64 aneurysms in 51 patients were treated. Among them, 18 aneurysms in 15 patients (4 ruptured aneurysms and 14 unruptured aneurysms) required an adjunctive technique using a TACTICS catheter. The methods of embolization were the double catheter technique (DCT) for five aneurysms, stent-assisted coiling (SAT) for seven, DCT + SAC for one, and balloon-assisted technique (BAT) for five aneurysms. Aneurysms were located in the anterior communicating artery (Acom A) in three cases, distal anterior cerebral artery (dACA) in one, middle cerebral artery (MCA) in five, internal carotid artery (ICA) in six, basilar artery (BA) in one, and vertebral artery (VA) in two cases. It was easy to access distal intracranial vessels using the TACTICS catheter. In all cases, guiding of the microcatheter, coiling, and stent placement were improved. There were no complications associated with using the TACTICS catheter., Conclusion: Compared with conventional DACs, distal intracranial vessels were more easily accessed using the TACTICS catheter. A TACTICS catheter is useful because DCT and SAC require sufficient operability of the microcatheter., Competing Interests: The authors declare no conflict of interest., (©2021 The Japanese Society for Neuroendovascular Therapy.)

A mixed aqueous solution of hydrophilic poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) and poly(acrylic acid) (PAAc) becomes cloudy under acidic conditions at room temperature. The pendant carboxylic acid groups in PAAc form hydrogen bonds with the ester and phosphate groups in PMPC. While the polymers aggregate under acidic conditions, neither one associate under basic conditions because of the deprotonation of the pendant carboxy groups in PAAc. We observed that the interpolymer complex formed from PMPC, and PAAc was dissociated in aqueous solutions with increasing temperature, which is an upper critical solution temperature behavior. With increasing temperature, the molecular motion increased to dissociate the interpolymer complex. The phase transition temperature increased with increasing polymer and salt concentrations, and with decreasing pH.

Ideal interaction-free magnetite nanoparticles were prepared, and their magnetic properties were measured to clarify the true nature of magnetic anisotropy of individual magnetite nanoparticles at the nanoscale and to analyze the shape, surface, and crystalline anisotropy contributions. Spherical (17.7 nm), cubic (10.6 nm), and octahedral-shaped magnetite nanoparticles with average sizes ranging from 7.6 to 23.4 nm were synthesized using solution techniques. Then, these nanoparticles were coated with silica at appropriate shell thicknesses to prepare magnetic interaction-free samples, and their noninteractive nature was confirmed through first-order reversal curve diagrams. For these well-isolated nanoparticles, remanent magnetizations of the hysteresis loops are just equal to a half of the saturation magnetization. This result clearly indicates that uniaxial magnetic anisotropy is predominant in each nanoparticle. In order to clarify the details of the uniaxial magnetic anisotropy, the analysis of blocking temperature-switching field distribution diagrams is constructed based on thermal decay curves of isothermal remanent magnetization at various applied fields. The obtained effective magnetic anisotropy constant K eff is distributed around 10-20 kJ/m 3 and has insignificant size dependence. This result seems inconsistent with the inverse proportion relation of K eff with size predicted for surface magnetic anisotropy. The theoretical calculation suggested that the crystalline magnetic anisotropy plays a major role in magnetic properties of the magnetite nanoparticles at lower temperatures. However, it should be noted that K eff seems slightly different for the different shapes. The above study indicates that control size, shape, and interparticle interactions is required to strictly discuss such delicate differences of magnetic anisotropy of individual magnetite nanoparticles for the design of thermal seeds for magnetic hyperthermia.

Among the four bioactive cationic homo-poly(amino acids) discovered in nature, two are mirror-image isomers of poly(2,4-diaminobutyric acid) (poly-Dab) whose biosynthesis has long been unexplained. Their structural analogy plausibly suggested that they could share a common biosynthetic pathway utilizing ε-poly(l-lysine) synthetase-like enzymology but with an unprecedented process for enantiomeric inversion of polymer building blocks. To investigate this possibility, we comparatively explored the biosynthesis of poly-l-Dab and its mirror-image isomer poly-d-Dab in Streptomyces celluloflavus USE31 and Streptoalloteichus hindustanus NBRC15115, respectively, through genome mining, genetic inactivation, and heterologous expression combined with biochemical assays. While they shared the same biosynthetic pathway, the poly-d-Dab biosynthetic gene cluster additionally harbored the racemase gene. The critical finding that poly-d-Dab synthetase, in contrast to the synthetase generating the l-isomer, selectively activated d-Dab through adenylation conclusively demonstrated that free diffusible d-Dab preactivationally generated by the racemase is directly activated to be incorporated into the polymer. Our study thus represents the first demonstration of the stereoselective biosynthesis of a nonribosomal peptide governed by adenylation activity for a d-amino acid other than alanine. In silico sequence comparison between poly-Dab synthetases allowed us to identify amino acid residues potentially responsible for the discrimination of Dab enantiomers. Our results will provide significant insight not only for the future discovery of novel bioactive cationic poly(amino acids) but also for the creation of designer nonribosomal peptides with d-configuration.

Microvascular decompression(MVD)is an effective treatment for hemifacial spasm and trigeminal neuralgia. However, there are few reports regarding its use with abducens nerve palsy. Here, we report the case of a 77-year-old male who presented with diplopia and was admitted to our facility for right abducens nerve palsy. MRI constructive interference in steady-state(CISS)showed that the right anterior inferior cerebellar artery(AICA)was curved in a posterior-superior direction at the beginning of the region of origin, causing compression of the root exit zone of the right abducens nerve. In addition, MRI showed an unruptured fusiform aneurysm(5.3mm×7.1mm)of the vertebral artery(VA)involving the posterior inferior cerebellar artery(PICA). However it was not related to abducens nerve paralysis. The aneurysm was successfully treated with an occipital artery-posterior inferior cerebellar artery(OA-PICA)bypass and clipping of the proximal VA and PICA origin. The position of the offending artery was moved using a Teflon ® felt fibrin glue and the retrosigmoid transcondylar fossa approach. The patient recovered from abducens nerve paralysis with no new neurological deficit. We emphasize the importance of choosing an appropriate approach as this increases the possibility of treating abducens nerve paralysis in a timely manner.

Trichodysplasia spinulosa-associated polyomavirus (TSPyV or human polyomavirus 8) was identified from patients with trichodysplasia spinulosa, a rare skin disease affecting the faces of immunocompromised patients. Like other polyomaviruses, the TSPyV genome encodes a large T antigen (LT). However, the expression and functions of TSPyV LT in infected cells remain largely unknown. In the present study, we cloned a full-length TSPyV LT cDNA from cells transfected with the full-length of TSPyV LT DNA. Transfection study using green fluorescence protein-tagged LT expression plasmids showed that TSPyV LT was expressed in the nucleus of transfected cells. Analysis of deletion mutants identified a nuclear localization signal in TSPyV LT. Recombinant TSPyV LT exhibited an ATPase activity. TSPyV LT has a chitinase-like domain; however, no chitinase activity was detected. Immunoprecipitation assays revealed that TSPyV LT bound to retinoblastoma 1, but not to p53 in transfected cells. Expression of TSPyV LT in NIH3T3 cells induced colony formation in soft agar, suggesting its transformation activity. These data indicate that TSPyV LT may be associated with the pathogenesis of trichodysplasia spinulosa, which is a hyperplasia of keratinocytes in inner hair follicles.

Background: Chronic total occlusion (CTO) of femoropopliteal artery (FP) continues to be a lesion subset where maintaining long-term patency after endovascular treatment is challenging. We evaluated the efficacy of cutting balloon angioplasty (CBA) for de novo FP-CTOs in patients with symptomatic lower limb ischemia., Methods: Seventy-three limbs of 67 symptomatic patients with de novo FP-CTOs successfully recanalized using CBA alone were enrolled in this study. Primary patency was defined as the absence of recurrent symptoms and no deterioration of the ankle-brachial index (ABI) >0.10 from the immediate postinterventional value., Results: The mean age was 73.5 ± 7.3 years, and 59.7% of patients had diabetes mellitus. Most lesions were classified as Trans-Atlantic Inter-Society Consensus II type C (n = 18; 24.7%) or type D (n = 44; 60.3%), with mean lesion and occluded lengths of 24.8 ± 11.4 and 17.8 ± 11.2 cm, respectively. No procedure-related adverse events occurred, except one distal embolization. The ABI significantly increased after intervention from 0.52 ± 0.12 to 0.80 ± 0.15 (P < 0.0001), with marked improvement in clinical symptoms (Rutherford stage: 2.7 ± 1.0 to 1.1 ± 1.2, P < 0.0001). The mean follow-up period was 31.2 ± 18.0 months, and the primary patency rates at 12 and 24 months were 75.3% and 60.6%, respectively. The independent predictive factors of failed patency were baseline hemoglobin A1c (P = 0.031, hazard radio [HR] 1.51 per 1%), occluded length ≥15 cm (P = 0.036, HR 2.90), and severe dissection (P = 0.033, HR 2.85). Vessel calcification and diameter did not affect primary patency., Conclusions: CBA is a feasible option for endovascular treatment of FP-CTOs. Diabetic status, occlusion length, and severe dissection after CBA are independent negative predictors of long-term patency., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Intramolecular photocyclization of stilbene derivatives (Mallory reaction) is one of the efficient methods for building polycyclic aromatic hydrocarbon (PAH) frameworks, and is also expected to be applicable to synthesis of fluorine-containing PAHs (F-PAHs). In this study, dibenzoanthracene-type ( 4a ) and benzoperylene-type ( 4b ) F-PAHs were synthesized using the Mallory reaction of the 1,4-distyrylbenzene-type π-conjugated molecule ( 3a ), which was prepared by addition-defluorination of available octafluorocyclopentene (OFCP) and aryllithium in three steps. The structure of 4a originating from π⁻π interaction was characterized by X-ray crystallographic analysis. The absorption maxima of UV-Vis spectra and emission maxima of photoluminescence spectra of the PAHs were positioned at a longer wavelength compared to those of the corresponding unsubstituted PAHs, presumably due to the electron-withdrawing nature of perfluorocyclopentene (PFCP) units. The effect of PFCP units in F-PAHs was also studied by time-dependent density functional theory (TD-DFT) calculation.

Cu(I), Ag(I), and Au(I) complexes of monophosphine or bisphosphine ligands based on the 3,3,4,4,5,5-hexafluorocyclopentene skeleton were synthesized and structurally characterized by X-ray crystallographic analysis. The electron-withdrawing nature of these polyfluorinated phosphines was experimentally revealed via UV/vis absorption studies and crystal structure analysis. Successful catalytic application of the Au(I) complex for alkyne hydration reactions was investigated.

Background: Functional mitral regurgitation (MR) can occur in patients with atrial fibrillation (AF) despite having preserved left ventricular (LV) systolic function. This MR is known as atrial functional MR. The aim of this study was to clarify the mechanism of atrial functional MR using real-time three-dimensional transesophageal echocardiography (3DTEE)., Methods: Sixty patients underwent transthoracic echocardiography and 3DTEE: 16 patients with AF and significant non-organic MR and preserved LV ejection fraction (>50%) constituted the AF-MR group, 20 patients with AF and no significant MR formed the AF-NSMR group, and 24 normal subjects comprised the control group., Results: The left atrial volume index was significantly larger in the AF-MR group (95±41ml/m 2 ) than in the AF-NSMR group (38±13ml/m 2 , p<0.05) or the control group (21±7ml/m 2 , p<0.05). The 3D annular circumference was significantly longer in the AF-MR group than in the AF-NSMR group. The annular-anterior leaflet coaptation angle was smaller in the AF-MR group than in the AF-NSMR group (11±6° vs. 18±9°, p<0.05). The annular-posterior leaflet coaptation angle was comparable between the two AF groups (26±12° vs. 28±10°), whereas the annular-posterior leaflet tip angle was larger in the AF-MR group than in the AF-NSMR group (59±13° vs. 44±11°, p<0.05). The posterior leaflet bending toward LV cavity was therefore significantly larger in the AF-MR group than in the AF-NSMR group (32±10° vs. 18±15°, p<0.05)., Conclusions: In patients with AF and significant functional MR occurring despite their preserved LV systolic function, the left atrium and mitral annulus were dilated and the anterior leaflet was flattened along the mitral annular plane, whereas the posterior leaflet was bent toward the LV cavity., (Copyright © 2017 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.)

Plasmablastic lymphoma (PBL) is a rare, highly aggressive subtype of non-Hodgkin lymphoma with plasma-cell differentiation occurring typically in immune-suppressed patients such as those with AIDS. This study reports the establishment and characterization of a new cell line, PBL-1, derived from a patient with AIDS-associated PBL. Morphological assessment of PBL-1 indicated plasma-cell differentiation with a CD20(-) CD38(+) CD138(+) immunophenotype and IgH/c-myc translocation. The cell line harbours Epstein-Barr virus, but a 52.7-kbp length defect was identified in its genome, resulting in no expression of viral microRNAs encoded in the BamHI-A Rightward Transcript region. Importantly, supplementation of culture medium with >5 ng/mL of interleukin-6 (IL-6) was required for PBL-1 growth. Starvation of IL-6 or addition of tocilizumab, an inhibitory antibody for the IL-6 receptor, induced apoptosis of PBL-1. Transduction of IL-6 into PBL-1 by lentivirus vector induced autologous growth without IL-6 supplementation of culture medium. These data indicate the IL-6 dependency of PBL-1 for proliferation and survival. mTOR inhibitors induced cell death effectively, suggesting mTOR in the IL-6 signalling pathway is a potential therapeutic target for PBL. This established PBL cell line will be a useful tool to further understand the pathophysiology of PBL and aid the future development of PBL treatment.

Competing Interests: Conflict of Interest: No conflict of interest was declared by the authors.

Objective: Epstein-Barr virus (EBV) encodes at least 25 pri-microRNAs (miRNAs) in two regions of its DNA genome, BART and BHRF. B95-8, an EBV reference strain, has a deletion in the BART region. However, no information is available on the deletions or mutations in the BART and BHRF regions in clinical samples of EBV-associated lymphoma., Results: Nine DNA fragments encoding miR-BARTs and two coding miR-BHRF1s were amplified by PCR from DNA samples extracted from 16 cases of EBV-associated lymphoma. All the PCR products were sequenced directly. DNA fragments encoding miR-BARTs and miR-BHRF1-1 were successfully amplified from all samples. An adenine-to-guanine mutation in the DNA fragment encoding miR-BART2-3p was detected in four of the 16 cases, and a cytosine-to-thymidine mutation in the DNA fragment encoding miR-BART11-3p was detected in one of the 16 samples. These mutations were not associated with any histological categories of lymphoma. In conclusion, mutations were rarely observed in the DNA encoding viral miRNAs in cases of lymphoma. This suggests that the DNA sequences of EBV-encoded miR-BARTs and miR-BHRF1-1 are conserved in EBV-associated lymphoma.

Sialidase cleaves sialic acids on the extracellular cell surface as well as inside the cell and is necessary for normal long-term potentiation (LTP) at mossy fiber-CA3 pyramidal cell synapses and for hippocampus-dependent spatial memory. Here, we investigated in detail the role of sialidase in memory processing. Sialidase activity measured with 4-methylumbelliferyl-α-d- N -acetylneuraminic acid (4MU-Neu5Ac) or 5-bromo-4-chloroindol-3-yl-α-d- N -acetylneuraminic acid (X-Neu5Ac) and Fast Red Violet LB was increased by high-K + -induced membrane depolarization. Sialidase activity was also increased by chemical LTP induction with forskolin and activation of BDNF signaling, non-NMDA receptors, or NMDA receptors. The increase in sialidase activity with neural excitation appears to be caused not by secreted sialidase or by an increase in sialidase expression but by a change in the subcellular localization of sialidase. Astrocytes as well as neurons are also involved in the neural activity-dependent increase in sialidase activity. Sialidase activity visualized with a benzothiazolylphenol-based sialic acid derivative (BTP3-Neu5Ac), a highly sensitive histochemical imaging probe for sialidase activity, at the CA3 stratum lucidum of rat acute hippocampal slices was immediately increased in response to LTP-inducible high-frequency stimulation on a time scale of seconds. To obtain direct evidence for sialic acid removal on the extracellular cell surface during neural excitation, the extracellular free sialic acid level in the hippocampus was monitored using in vivo microdialysis. The free sialic acid level was increased by high-K + -induced membrane depolarization. Desialylation also occurred during hippocampus-dependent memory formation in a contextual fear-conditioning paradigm. Our results show that neural activity-dependent desialylation by sialidase may be involved in hippocampal memory processing., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)

Epstein-Barr virus (EBV) encodes 49 microRNAs (miRNAs) in the BART and BHRF1 regions of its genome. Although expression profiles of EBV-encoded miRNAs have been reported for EBV-positive cell lines and nasopharyngeal carcinoma, to date there is little information about total miRNA expression, including cellular and viral miRNAs, in the primary tumors of EBV-associated B-lymphoproliferative disorders. In this study, next-generation sequencing and quantitative real-time reverse transcription-PCR were used to determine the expression profiles of miRNAs in EBV-infected cell lines and EBV-associated B-cell lymphomas, including AIDS-related diffuse large B-cell lymphoma (DLBCL), pyothorax-associated lymphoma, methotrexate-associated lymphoproliferative disorder, EBV-positive DLBCL of the elderly, and Hodgkin lymphoma. Next-generation sequencing revealed that EBV-encoded miRNAs accounted for up to 34% of total annotated miRNAs in these cases. Expression of three miR-BHRF1s was significantly higher in AIDS-related DLBCL and pyothorax-associated lymphoma compared with methotrexate-associated lymphoproliferative disorder and EBV-positive DLBCL of the elderly, suggesting the association of miR-BHRF1s expression with latency III EBV infection. Heat map/clustering analysis of expression of all miRNAs, including cellular and EBV miRNAs, by next-generation sequencing demonstrated that each EBV tumor, except methotrexate-associated lymphoproliferative disorder, formed an isolated cluster. Principal component analysis based on the EBV-encoded miRNA expression showed that each EBV tumor formed a distinguished cluster, but AIDS-related DLBCL and pyothorax-associated lymphoma formed larger clusters than other tumors. These data suggest that expression of miRNAs, including EBV-encoded miRNAs, is associated with the tumor type and status of virus infection in these tumors., (© 2017 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

An efficient method for preparing unsymmetrically multisubstituted siloles is described. The reaction of 1-hydrosilyl-4-silyl-1,3-enynes with diisobutylaluminum hydride (DIBAL-H) gave multisubstituted siloles in good to high yields. This method could be applied to the synthesis of benzosiloles using 2-hydrosilyl-1-(silylethynyl)benzenes as substrates. The silole formation was also promoted even by a substoichiometric amount of DIBAL-H. The reaction provides a straightforward method to prepare siloles and benzosiloles.

JC virus (JCV) is a DNA virus causing progressive multifocal leukoencephalopathy (PML) in immunodeficient patients. In the present study, 22 genetic quasispecies with more than 1.5% variant frequency were detected in JCV genomes from six clinical samples of PML by next-generation sequencing. A mutation from A to C at nucleotide (nt) 3495 in JCV Mad1 resulting in a V-to-G amino acid substitution at amino acid (aa) position 392 of the large T antigen (TAg) was identified in all six cases of PML at 3% to 19% variant frequencies. Transfection of JCV Mad1 DNA possessing the V392G substitution in TAg into IMR-32 and human embryonic kidney 293 (HEK293) cells resulted in dramatically decreased production of JCV-encoded proteins. The virus DNA copy number was also reduced in supernatants of the mutant virus-transfected cells. Transfection of the IMR-32 and HEK293 cells with a virus genome containing a revertant mutation recovered viral production and protein expression. Cotransfection with equal amounts of wild-type genome and mutated JCV genome did not reduce the expression of viral proteins or viral replication, suggesting that the mutation did not have any dominant-negative function. Finally, immunohistochemistry demonstrated that TAg was expressed in all six pathological samples in which the quasispecies were detected. In conclusion, the V392G amino acid substitution in TAg identified frequently in PML lesions has a function in suppressing JCV replication, but the frequency of the mutation was restricted and its role in PML lesions was limited., Importance: DNA viruses generally have lower mutation frequency than RNA viruses, and the detection of quasispecies in JCV has rarely been reported. In the present study, a next-generation sequencer identified a JCV quasispecies with an amino acid substitution in the T antigen in patients with PML. In vitro studies showed that the mutation strongly repressed the expression of JC viral proteins and reduced the viral replication. However, because the frequency of the mutation was low in each case, the total expression of virus proteins was sustained in vivo. Thus, JC virus replicates in PML lesions in the presence of a mutant virus which is able to repress virus replication., (Copyright © 2016 American Society for Microbiology.)

Growing evidence suggests persistent mitochondrial permeability transition pore (mPTP) opening is a key pathophysiological event in cell death underlying a variety of diseases. While it has long been clear the mPTP is a druggable target, current agents are limited by off-target effects and low therapeutic efficacy. Therefore identification and development of novel inhibitors is necessary. To rapidly screen large compound libraries for novel mPTP modulators, a method was exploited to cryopreserve large batches of functionally active mitochondria from cells and tissues. The cryopreserved mitochondria maintained respiratory coupling and ATP synthesis, Ca 2+ uptake and transmembrane potential. A high-throughput screen (HTS), using an assay of Ca 2+ -induced mitochondrial swelling in the cryopreserved mitochondria identified ER-000444793, a potent inhibitor of mPTP opening. Further evaluation using assays of Ca 2+ -induced membrane depolarisation and Ca 2+ retention capacity also indicated that ER-000444793 acted as an inhibitor of the mPTP. ER-000444793 neither affected cyclophilin D (CypD) enzymatic activity, nor displaced of CsA from CypD protein, suggesting a mechanism independent of CypD inhibition. Here we identified a novel, CypD-independent inhibitor of the mPTP. The screening approach and compound described provides a workflow and additional tool to aid the search for novel mPTP modulators and to help understand its molecular nature., Competing Interests: T.B., S.L., M.K., K.M., N.H., M.R., B.P., Y.S., H.F. and J.S. are employees of Eisai.

Background: Primary effusion lymphoma is a rare distinct large B-cell neoplasm that is associated with Kaposi's sarcoma-associated herpesvirus (KSHV) infection. Over recent years, 9 KSHV-positive/Epstein-Barr virus (EBV)-negative PEL cell lines have been established., Methods: Tumor cells were collected from the pleural effusion of a 49-year-old male with AIDS. Cells were grown in RPMI1640 culture medium supplemented with 10 % fetus bovine serum. Single cell cloning was performed successfully by a limiting dilution method in a 96-well plate. The cell line obtained was designated SPEL., Results: SPEL cells showed gourd-shaped morphology with a polarized nucleus, expressing CD38, CD138, and Blimp-1, but not B cell markers such as CD19 and CD20. Polymerase chain reaction analysis revealed that SPEL cells were positive for KSHV but negative for EBV. Tetradecanoylphorbol acetate induced expression of KSHV lytic proteins and the production of KSHV particles in SPEL cells. Subcutaneous inoculation of SPEL cells into severe combined immunodeficiency mice resulted in the formation of solid tumors. Next-generation sequencing revealed the 138 kbp genome sequence of KSHV in SPEL cells. Suberic bishydroxamate, a histone deacetylase inhibitor, induced the expression of KSHV-encoded lytic proteins and cell death in SPEL cells., Conclusions: A new KSHV-positive and EBV-negative PEL cell line, SPEL was established. This cell line may contribute to furthering our understanding of the pathogenesis of PEL and KSHV infection.

Background: Vero cells are used in laboratories for the isolation of viruses and the production of vaccines. Recently, the sequence of simian endogenous retrovirus (SERV) was identified in Vero cells (SERVagm-Vero), with homology to exogenously transmitted, pathogenic simian retroviruses (SRVs). Although SERVagm-Vero was shown to be noninfectious to human cells in vitro, SERV infection in humans is controversial. In this study, we evaluated the status of SERV infection in humans by detecting the viral genome in clinicopathological samples and commercialized vaccines, and with a serological survey of the Japanese general population., Methods: Real-time polymerase chain reaction (PCR) and reverse transcription-PCR were used to detect the SERVagm-Vero genome. We also examined the seroprevalence of SERV in 1000 individuals in the Japanese population with an enzyme-linked immunoabsorbent assay (ELISA) using mixed SERVagm-Vero gag, pol, and env proteins as antigens., Results: Real-time PCR failed to detect SERVagm-Vero genomic fragments in 783 human clinicopathological samples, and all 13 human cell lines tested were negative for the SERVagm-Vero genome. Thirteen commercialized vaccines, including five Vero-based vaccines, were also negative for the SERVagm-Vero genome on real-time PCR and reverse transcription-PCR. Eight (0.8%) were seropositive on ELISA, and western blotting showed that all eight sera contained anti-pol antibodies. All SERV-seropositive individuals were born before 1965, suggesting that SERV infection in Japan might not be associated with vaccine, because more than 90% of Japanese children born from 1964 to 2012 have received live poliovirus vaccines containing virus produced in Vero cells since the 1980s., Conclusion: We have confirmed that the vaccines we use today are free of SERVagm-Vero. Moreover, SERV infection in humans is very rare and unlikely to be associated with vaccines in the Japanese general population., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Symptomatic extensive spinal extradural meningeal cyst (SEMC) developing after traumatic brachial plexus injury (TBPI) is rare. We discuss the mechanism of extensive SEMC development, surgical strategies, and preventive measures against SEMC after TBPI., Case Description: A 58-year-old man with TBPI 16 years previously developed spastic paraparesis of the lower limbs, sensory disturbance below the periumbilical level, and dysfunction of bladder and bowel over 2 years. The patient couldn't walk and was wheelchair bound. Magnetic resonance imaging (MRI) revealed an extensive multilocular extradural cyst posterior to the spinal cord ranging from the C4 to Th6 level, associated with severe spinal cord compression. On constructive interference in steady-state MRI, the cyst was divided, with many septa, and extended to the root sleeves. During the operation, transdural communication sites of cerebrospinal fluid (CSF) into the cyst were revealed at C5/6, C6/7, and C7/Th1 levels around the nerve root sleeves. Treatment involved unroofing of the cyst wall and closure of the transdural CSF communication without cyst removal. Autologous muscle pieces were placed over the defect to close the transdural communication. Two weeks postoperatively, MRI showed decreased cyst size and reduced spinal cord compression, and the patient could walk without support. It was thought that the patient's daily lifting of heavy weights at work and an excessive exercise regimen increased CSF pressure and cyst size after TBPI., Conclusion: For patients with TBPI, it is necessary to prevent greater CSF pressure and to perform long-term follow-up MRI after injury., (Copyright © 2016 Elsevier Inc. All rights reserved.)

A novel system for simultaneous detection of pathogenic bacteria and fungi in pathological samples was developed using a real-time polymerase chain reaction (PCR) system. This system, designated the "multi-microbial real-time PCR", has the potential to simultaneously detect 68 bacterial and 9 fungal species in a 96-well plate format. All probe-primer sets were designed to produce amplicons smaller than 210 bp using formalin-fixed paraffin-embedded samples as input. The specificity and sensitivity of each probe-primer set were tested against DNA extracted from pure cultures of specific pathogens. The multi-microbial real-time PCR system revealed profiles of microorganism infection in lung samples collected at autopsy from 10 patients with acquired immunodeficiency syndrome. Staphylococcus aureus was the most common microbe detected (n=8), but with low copy numbers. High copy numbers of Pseudomonas aeruginosa were detected in the lung samples with abscess (n=6). Enterococcus faecium (n=6), Elizabethkingia meningoseptica (n=4), and Candida albicans (n=4) were also frequently detected. In addition, a latent infection of Mycobacterium tuberculosis was detected in one case of pneumonia. In conclusion, this multi-microbial real-time PCR system can be useful for detecting bacteria and fungi in pathological specimens from patients with uncertain diagnoses.

Purpose: In this study, we examined the hypothesis that the preferential conduction property along left lateral ridge (LLR) might affect the arrhythmogenicity of left pulmonary veins (LPVs)., Methods: The study population included 40 consecutive AF patients. Radiofrequency energy (RF) was sequentially delivered along the LLR from a lower to upper manner during postero-lateral CS pacing during an isoproterenol infusion., Results: The conduction time during pacing from the CS was significantly prolonged during radiofrequency (RF) deliveries (before vs. after, upper; 91±26ms vs. 127±38ms, p<0.001, lower; 86±21ms vs. 103±22ms, p<0.001). Remarkable prolongation of more than 30ms was observed in 19 of 40 patients (48%) (both LPVs, 6; only the upper LPVs, 12; and only the lower LPV, 1). Sites with a remarkable prolongation were observed at the carina between the LPVs,[4] anterior site of the upper LPV carina,[10] anterior wall of the lower LPV,[3] and bottom of the lower LPVs [2] Thirty-three arrhythmogenic foci (AMF) from the LPVs were observed in 23/40 patients (56%). The conduction time during pacing from the LPVs during the RF delivery was significantly longer in the patients with AMF from the upper LPV than in those patients without (107±36ms vs. 146±40ms, p<0.01)., Conclusion: The LLR includes the preferential conduction properties between the CS and LPVs, and the observation of the serial changes during the RF delivery could provide us information about the LPVs arrhythmogenicity.

Kaposi sarcoma and primary effusion lymphoma cells are infected with Kaposi sarcoma-associated herpesvirus (KSHV), predominantly in the latent form, and KSHV replication is observed rarely. Angiogenesis plays a crucial role in the pathogenesis of both Kaposi sarcoma and primary effusion lymphoma. In this study, we found that fumagillin, a potent angiogenesis inhibitor, induced replication of KSHV in primary effusion lymphoma cell lines. The transcript and protein product of replication transcriptional activator (RTA) were induced by 1-10 μM fumagillin at 24 and 48 h, respectively. Western blot analysis demonstrated that 10 μM fumagillin induced not only RTA expression but also other KSHV-encoded lytic proteins. A real-time PCR array detecting KSHV gene expression demonstrated that the expression profiles of KSHV induced by fumagillin were similar to those induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), but the amounts of each transcript were lower than those induced by TPA. Finally, real-time PCR demonstrated an increase in that viral DNA copy number per cell in fumagillin-stimulated primary effusion lymphoma cell lines, indicating replication of KSHV. In addition to TPA, 10 μM fumagillin resulted in growth inhibition of primary effusion lymphoma cell lines. These observations suggest that an angiogenesis inhibitor is an agent with potent effects on cell growth and KSHV reactivation in primary effusion lymphoma cells., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Trichodysplasia spinulosa-associated polyomavirus (TSV) was identified in, and shown to be the probable cause of, trichodysplasia spinulosa, a rare skin disease. To date, serological analyses have revealed that TSV infection is common among adults in the general population of Europe and Australia. However, there have been no reports of TSV in Asia., Objective: To study the prevalence of TSV infection in Japan., Study Design: TSV-VP1 expressed in a recombinant baculovirus expression system in an insect cell line, Tn5, self-assembled into virus-like particles. Overall, 1000 serum samples were examined by enzyme-linked immunosorbent assays using virus-like particles of TSV as antigen. Participants ranged in age from 0 to 94 years., Results: Overall, 629 of 1000 serum samples (62.9%) were positive for anti-TSV antibodies. The seropositive rate increased with age and the seroprevalence of TSV significantly increased from 17.1% (25/146) in children aged from 0 to 4 years to 78.7% (472/600) in adults aged over 20 years (odds ratio = 0.056, 95% confidence interval = 0.035-0.900, P = 0.000, Chi-squared test). TSV seropositivity was not different between sera obtained in 1980 and 2012, and was not associated with sex. Competitive assay demonstrated that TSV antibodies did not cross-react with BK virus or Merkel cell polyomavirus., Conclusions: These results provide evidence that TSV circulates widely in the Japanese population, with primary exposure occurring mainly at early childhood, similar to that previously reported in other countries., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: The features of intrinsic ganglionated plexi (GP) in both atria after extensive pulmonary vein isolation (PVI) and their clinical implications have not been clarified in patients with atrial fibrillation (AF)., Objective: The purpose of this study was to assess the features of GP response after extensive PVI and to evaluate the relationship between GP responses and subsequent AF episodes., Methods: The study population consisted of 216 consecutive AF patients (104 persistent AF) who underwent an initial ablation. We searched for the GP sites in both atria after an extensive PVI., Results: GP responses were determined in 186 of 216 patients (85.6%). In the left atrium, GP responses were observed around the right inferior GP in 116 of 216 patients (53.7%) and around the left inferior GP in 57 of 216 (26.4%). In the right atrium, GP responses were observed around the posteroseptal area: inside the CS in 64 of 216 patients (29.6%), at the CS ostium in 150 of 216 (69.4%), and in the lower right atrium in 45 of 216 (20.8%). The presence of a positive GP response was an independent risk factor for AF recurrence (hazard ratio 4.04, confidence interval 1.48-11.0) in patients with paroxysmal, but not persistent, AF. The incidence of recurrent atrial tachyarrhythmias in patients with paroxysmal AF with a positive GP response was 51% vs 8% in those without a GP response (P = .002)., Conclusion: The presence of GP responses after extensive PVI was significantly associated with increased AF recurrence after ablation in patients with paroxysmal AF., (Copyright © 2015 Heart Rhythm Society. Published by Elsevier Inc. All rights reserved.)

We report a case of Merkel cell carcinoma (MCC) on the dorsal aspect of the right middle finger associated with multiple squamous cell carcinomas (SCC) possibly arising in chronic radiation dermatitis of the hand of an 80-year-old surgeon. In spite of resection of the primary lesion and right axillary lymph nodes, he died of the tumor 5 months after the first visit. Cutaneous and lymph node lesions of MCC were negative for Merkel cell polyoma virus (MCPyV) by immunostaining using monoclonal antibody (CM2B4) and anti-large T antigen of MCPyV polyclonal antibody, and real-time polymerase chain reaction. Several differences in clinicopathological findings have been found between MCPyV-positive cases and negative ones. Several authors have reported that MCPyV-negative cases have a worse prognosis than MCPyV-positive ones. Furthermore, in cases of MCC associated with SCC, most tumors have been reported to be MCPyV-negative. We should pay more attention to the relationship between the carcinogenesis of MCC and ionizing irradiation., (© 2014 Japanese Dermatological Association.)

Background: The mechanism of persistent atrial fibrillation (AF) is multifactorial, and arrhythmogenic foci (AMF) might be involved in the occurrence of persistent AF. In this study, we examined the electrophysiological features of AMF during and immediately after ablation, and evaluated the relationship between the presence and number of residual AMF on the risk of AF recurrence after a vigorous sinus rhythm restoration ablation in patients with long-standing persistent AF., Methods: The study consisted of 117 consecutive patients with persistent AF who underwent catheter ablation (CA). We performed direct cardioversion to restore sinus rhythm before the pulmonary vein (PV) isolation and at the end of the CA. Then we evaluated the features of the AMF inducible with isoproterenol and the pacing-based AF inducibility., Results: After the completion of ablation, AF could still be induced in 37 of 117 patients (31.6%). Spontaneous PV AMF during CA were observed in 104 of 117 patients (91%), and non-PV AMF in 63 of 117 (54%). Residual non-PV AMF were significantly associated with the pacing-based AF inducibility and an enlarged left atrial volume. In the multivariate analysis, the AF duration (1.01 [range, 1.00-1.02] months; P = 0.012), left atrial volume (1.01 [range, 1.01-1.02] mm; P = 0.006), and residual AMF (3.95 [range, 1.32-11.8] yes, no; P = 0.004) were independent risk factors for recurrent AF., Conclusions: Residual AMF are associated with an increased long-term AF recurrence after sinus rhythm restoration ablation for long-standing persistent AF., (Copyright © 2014 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.)

We describe a fatal case of drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome with human herpesvirus-6B (HHV-6B)-associated lymphadenitis and virus-associated hemophagocytic syndrome triggered by an over-the-counter medication to treat respiratory and influenza-like symptoms. Histologically, the structure of the lymph node was disrupted with infiltration of large lymphocytes carrying intranuclear acidophilic inclusion bodies. Immunohistochemistry and real-time PCR analysis revealed that these large lymphocytes were positive for HHV-6B. Numerous HHV-6 particles were detected in the inclusion body of the lymphocytes by electron microscopy. Interestingly, immunohistochemistry revealed that HHV-6B-infected cells in the lymph node were CD3(+), CD4(+), CD25(+), and FoxP3(+) T cells, indicating a phenotypic resemblance to regulatory T-cells. This case provides direct evidence of HHV-6 infection in CD25(+)/FoxP3(+) T cells in a case of acute lymphadenitis of DRESS syndrome, suggesting a significant role of HHV-6 infection of regulatory T-cells in the pathogenesis of DRESS syndrome., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objectives: To evaluate the feasibility and safety of a virtual 3-Fr system [5-Fr sheathless-guiding catheter (GC)] for percutaneous coronary intervention (PCI)., Background: The use of miniaturized devices for PCI is gaining popularity because of increased patient comfort and decreased risk of access site complications., Methods: From July 2010 to December 2012, consecutive patients who underwent elective PCI (planned or ad hoc PCI) at our hospital were enrolled. PCI using the virtual 3-Fr system was attempted as our initial strategy, unless a 6-Fr or larger GC was considered to be suitable [lesions with heavy calcification, large (>2 mm) side branches, or chronic total occlusion]., Results: Five hundred sixty-six patients underwent elective PCI during the study period, and 132 patients who met the criteria underwent PCI using the virtual 3-Fr system. Procedures using the virtual 3-Fr system were successful in 126 patients (95%); 111 (84%) were performed using the transradial approach, 110 (83%) were ad hoc procedures, and 45 (31%) were complex coronary lesions (type B2 or C). Six patients required conversion to a conventional 5- or 6-Fr sheath and catheter system. No intraoperative complications occurred, and radial artery patency was achieved in all patients who underwent transradial procedures., Conclusions: PCI using the virtual 3-Fr system is a feasible and viable alternative to conventional procedures that use a sheath and GC in appropriately selected patients. This small-caliber system may minimize endovascular trauma, particularly during transradial coronary procedures., (© 2014 Wiley Periodicals, Inc.)

Trichodysplasia spinulosa-associated polyomavirus (TSV) was identified in a seven-month-old girl with myocarditis. The number of TSV genomes detected was higher in the heart than in the other organs. The full-length TSV genome was cloned from the heart. This suggests a possible role of TSV infection in the pathogenesis of myocarditis in infants.

π-Conjugated polymers based on indigo unit were prepared. Dehalogenative polycondensation of N -hexyl-6,6'-dibromoindigo with a zerovalent nickel complex gave a homopolymer, P(HexI) , in 77% yield. Copolymer of N -hexyl-indigo and pyridine, P(HexI-Py) , was also prepared in 50% yield. P(HexI) showed good solubility in organic solvents, whereas P(HexI-Py) was only soluble in acids such as HCOOH. The weight-average molecular weights ( M w ) of P(HexI) and P(HexI-Py) were determined to be 10,000 and 40,000, respectively, by a light scattering method. Pd-catalyzed polycondensation between 6,6'-dibromoindigo with N -BOC (BOC = t -butoxycarbonyl) substituents and a diboronic compound of 9,9-dioctylfluorene afforded the corresponding alternating copolymer, P(BOCI-Flu) , as a deep red solid in 98% yield. P(BOCI-Flu) was soluble in N -methyl-2-pyrroridone and showed an M w of 29,000 in GPC analysis. Treatment of P(BOCI-Flu) with CF₃COOH smoothly led to a BOC-deprotection reaction to give an insoluble deep green polymer, P(I-Flu) , in a quantitative yield. Diffuse reflectance spectra of powdery P(BOCI-Flu) and P(I-Flu) showed peaks at about 580 nm and 630 nm, respectively, which are thought to originate from the indigo unit.

Background: Severe fever with thrombocytopenia syndrome (SFTS) is caused by SFTS virus (SFTSV), a novel bunyavirus reported to be endemic in central and northeastern China. This article describes the first identified patient with SFTS and a retrospective study on SFTS in Japan., Methods:  Virologic and pathologic examinations were performed on the patient's samples. Laboratory diagnosis of SFTS was made by isolation/genome amplification and/or the detection of anti-SFTSV immunoglobulin G antibody in sera. Physicians were alerted to the initial diagnosis and asked whether they had previously treated patients with symptoms similar to those of SFTS., Results: A female patient who died in 2012 received a diagnosis of SFTS. Ten additional patients with SFTS were then retrospectively identified. All patients were aged ≥50 years and lived in western Japan. Six cases were fatal. The ratio of males to females was 8:3. SFTSV was isolated from 8 patients. Phylogenetic analyses indicated that all of the Japanese SFTSV isolates formed a genotype independent to those from China. Most patients showed symptoms due to hemorrhage, possibly because of disseminated intravascular coagulation and/or hemophagocytosis., Conclusions: SFTS has been endemic to Japan, and SFTSV has been circulating naturally within the country.

Sialidase removes sialic acid from sialoglycoconjugates and plays crucial roles in many physiological and pathological processes. Various human cancers express an abnormally high level of the plasma membrane-associated sialidase isoform.Visualization of sialidase activity in living mammalian tissues would be useful not only for understanding sialidase functions but also for cancer diagnosis. However, since enzyme activity of mammalian sialidase is remarkably weak compared with that of bacterial and viral sialidases, it has been difficult to detect sialidase activity in mammalian tissues. We synthesized a novel benzothiazolylphenol-based sialic acid derivative (BTP-Neu5Ac) as a fluorescent sialidase substrate. BTP-Neu5Ac can visualize sialidase activities sensitively and selectively in acute rat brain slices. Cancer cells implanted orthotopically in mouse colons and human colon cancers (stages T3-T4) were also clearly detected with BTP-Neu5Ac. The results suggest that BTP-Neu5Ac is useful for histochemical imaging of sialidase activities.

Glucagon-like peptide 2 (GLP-2) is an intestinotropic peptide that binds to GLP-2 receptor (GLP-2R), a class-B G protein-coupled receptor. The GLP-2R antagonist GLP-2(3-33) has relatively high partial agonistic activity, and there are as yet no ideal known potent GLP-2R antagonists. We therefore prepared several truncated forms of human GLP-2 and characterized them by binding and reporter assays to find antagonists more potent than GLP-2(3-33). We found that GLP-2(11-33) was the most potent orthosteric GLP-2R antagonist, with binding activity almost equal to those of GLP-2 and GLP-2(3-33) and weaker intrinsic agonistic activity than GLP-2(3-33). GLP-2(11-33) retained weak agonistic activity toward human, cynomolgus monkey, dog, and Syrian hamster GLP-2Rs. However, it had no agonistic activity toward rat GLP-2R. GLP-2(11-33) potentiated the agonistic activity of an ago-allosteric modulator of GLP-2R, compound 1 (N-[1-(2,5-dichlorothiophen-3-yl)-2-(phenylsulfanyl)ethylidene]hydroxylamine), synergistically toward human GLP-2R. In the case of rat GLP-2R, GLP-2(11-33) decreased the agonistic activity of compound 1, although GLP-2 and GLP-2(3-33) increased this activity additively. These findings suggest that the binding sites of the ago-allosteric modulator and GLP-2 overlap, at least in rat GLP-2R. GLP-2(11-33) is a novel, useful tool for analyzing the mode of action of agonists and ago-allosteric modulators of GLP-2R.

Kaposi's sarcoma-associated herpesvirus (KSHV), the etiologic agent of Kaposi's sarcoma, causes malignancies frequently in patients with acquired immunodeficiency syndrome. In the United States and Europe, KSHV infection is common among men who have sex with men. However, the seroprevalence of KSHV among men who have sex with men in Japan is unknown. In the present study, the seroprevalence of KSHV was investigated among 230 men who have sex with men and 400 age- and area of residence-matched men (controls) using a mixed-antigen (KSHV-encoded K8.1, open reading frame 59, 65, and 73 proteins) enzyme-linked immunosorbent assay and an immunofluorescence assay. Among the Japanese men who have sex with men, serological assays revealed that 27 (11.7%) were seropositive for KSHV; 20 (5%) of the men in the control group were also KSHV seropositive. The seroprevalence of KSHV among men who have sex with men was significantly higher than in the control group (odds ratio = 2.52, 95% confidence intervals = 1.38-4.62, P = 0.0019, Chi-square test). Infection with the human immunodeficiency virus, Treponema pallidum, or hepatitis B and C virus did not correlate with KSHV infection. Furthermore, the association of KSHV seropositivity with specific sexual activities was not statistically significant. In conclusion, a higher KSHV seroprevalence was found among Japanese men who have sex with men than among the controls, suggesting that the circulation of KSHV infection is more efficient among men who have sex with men in Japan than among men who do not engage in such sexual activities., (Copyright © 2013 Wiley Periodicals, Inc.)

Background: Merkel cell polyomavirus (MCPyV), human polyomavirus-6 (HPyV6), and human polyomavirus-7 (HPyV7) were identified as viruses shed from the skin. Serological analysis revealed that these viruses are common among the general population. However, there is little information about the presence of MCPyV, HPyV6, and HPyV7 in the sera and tissues of immunocompromised individuals. The aims of this study are to know if immune status affects the presence of MCPyV, HPyV6, and HPyV7 in the serum, and to reveal the presence of these viruses in diseased tissues of unknown etiology., Methods: Sera from HIV-1-positive and -negative patients were examined by real-time PCR and nested PCR detecting MCPyV, HPyV6 and HPyV7. In addition, diseased tissue samples of unknown etiology were examined., Results: Nine out of 23 serum samples (39.1%) from HIV-1-positive patients who had not received anti-retroviral therapy were positive for MCPyV, which is significantly higher than HIV-1-negative patients (6/110, 5.5%, P<0.01, Chi-square test). MCPyV DNA was detected in tissue samples of Merkel cell carcinoma (22/30 [73%]), encephalitis (4/19 [21%]), pneumonia (3/17 [18%]), and myocarditis (8/14 [57%]). With the exception of Merkel cell carcinoma samples, MCPyV-positive tissues showed low copy numbers of MCPyV DNA by real-time PCR and no expression of the MCPyV large T antigen by immunohistochemistry. HPyV6 and HPyV7 were rarely detected in serum and tissue samples., Conclusions: These results suggest that MCPyV viremia is associated with host immunity, and that circulation of HPyV6 and HPyV7 in the serum is rare.

Glucagon-like peptide 2 (GLP-2) is an intestinotropic peptide that binds to GLP-2 receptor (GLP- 2R), a class-B G protein-coupled receptor (GPCR) coupled with Gα(s). Few small-molecule agonists had been reported for class-B GPCRs, but we recently reported the first scaffold compounds of ago-allosteric modulators for human GLP-2R. Methyl 2-{[(2Z)-2-(2,5-dichlorothiophen- 3-yl)-2-(hydroxyimino)ethyl]sulfanyl}benzoate (compound 1) and its de-esterified derivative (compound 2) induced placental alkaline phosphatase (PLAP) activity in HEK293 cells overexpressing human GLP-2R and PLAP driven by cAMP response element. In this study, we observed that rat, Syrian hamster, and dog GLP-2Rs also responded to compounds 1 and 2 in the same reporter system. However, no agonistic activity of the compounds toward mouse GLP-2R was detected. Mutagenesis studies showed that mutant human GLP-2Rs with Pro392Leu substitution of mouse GLP-2R for human GLP-2R amino acid residues nullified the PLAP activity of compound 2, although these mutant receptors responded to GLP-2. This finding suggests that the Pro392 residue of human GLP-2R is essential for the agonistic activity of compound 2.

A large thrombus load on the culprit coronary artery of patients with acute myocardial infarction (MI) is associated with increased procedural complications and adverse coronary events following angioplasty. This case series describes effective removal of large, occlusive thrombi in acute MI via direct aspiration using a standard 6 Fr guide catheter, following failed conventional catheter aspiration. This procedure is a simple and rapid alternative for challenging thrombi-containing coronary lesions when current thrombectomy catheters fail.

Glucagon-like peptide 2 (GLP-2) is an intestinotropic peptide that binds to GLP-2 receptor (GLP-2R), a class-B G protein-coupled receptor (GPCR). Few synthetic agonists have been reported so far for class-B GPCRs. Here, we report the first scaffold compounds of ago-allosteric modulators for human GLP-2R, derived from methyl 2-{[(2Z)-2-(2,5-dichlorothiophen-3-yl)-2-(hydroxyimino)ethyl]sulfanyl}benzoate (compound 1)., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Purpose: To describe a novel technique using an antegrade wire in a retrograde microcatheter advanced through a transcollateral vessel for recanalization of challenging infrapopliteal chronic total occlusions., Technique: A 75-year-old diabetic man presented with critical limb ischemia manifested as nonhealing ulcers on the toes. Baseline angiography revealed a blunt, long, total occlusion of the anterior tibial artery. A retrograde microcatheter was advanced over a guidewire tracking the collateral channel from the planter artery. Antegrade and retrograde microcatheters were aligned inside the occluded lesion. An antegrade wire was then advanced further down through the retrograde microcatheter. Final angiography of the anterior tibial artery following balloon dilation demonstrated a satisfactory result, without evidence of significant residual stenoses or flow-limiting dissections. Complete wound healing was achieved at 3 weeks., Conclusion: This alternative wire method may be useful when traditional interventional approaches are unfeasible.