Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells

K&ouml,nig, Alexander,Yang, Jaewon,Jo, Eunji,Park, Kyu Ho Paul,Kim, Hyun,Than, Thoa Thi,Song, Xiyong,Qi, Xiaoxuan,Dai, Xinghong,Park, Soonju,Shum, David,Ryu, Wang-Shick,Kim, Jung-Hee,Yoon, Seung Kew,P Elsevier 2019 Journal of hepatology Vol.71 No.2

<P><B>Background & Aims</B></P> <P>As hepatitis B virus (HBV) spreads through the infected liver it is simultaneously secreted into the blood. HBV-susceptible <I>in vitro</I> infection models do not efficiently amplify viral progeny or support cell-to-cell spread. We sought to establish a cell culture system for the amplification of infectious HBV from clinical specimens.</P> <P><B>Methods</B></P> <P>An HBV-susceptible sodium-taurocholate cotransporting polypeptide-overexpressing HepG2 cell clone (HepG2-NTCPsec+) producing high titers of infectious progeny was selected. Secreted HBV progeny were characterized by native gel electrophoresis and electron microscopy. Comparative RNA-seq transcriptomics was performed to quantify the expression of host proviral and restriction factors. Viral spread routes were evaluated using HBV entry- or replication inhibitors, visualization of viral cell-to-cell spread in reporter cells, and nearest neighbor infection determination. Amplification kinetics of HBV genotypes B-D were analyzed.</P> <P><B>Results</B></P> <P>Infected HepG2-NTCPsec+ secreted high levels of large HBV surface protein-enveloped infectious HBV progeny with typical appearance under electron microscopy. RNA-seq transcriptomics revealed that HBV does not induce significant gene expression changes in HepG2-NTCPsec+, however, transcription factors favoring HBV amplification were more strongly expressed than in less permissive HepG2-NTCPsec−. Upon inoculation with HBV-containing patient sera, rates of infected cells increased from 10% initially to 70% by viral spread to adjacent cells, and viral progeny and antigens were efficiently secreted. HepG2-NTCPsec+ supported up to 1,300-fold net amplification of HBV genomes depending on the source of virus. Viral spread and amplification were abolished by entry and replication inhibitors; viral rebound was observed after inhibitor discontinuation.</P> <P><B>Conclusions</B></P> <P>The novel HepG2-NTCPsec+ cells efficiently support the complete HBV life cycle, long-term viral spread and amplification of HBV derived from patients or cell culture, resembling relevant features of HBV-infected patients.</P> <P><B>Lay summary</B></P> <P>Currently available laboratory systems are unable to reproduce the dynamics of hepatitis B virus (HBV) spread through the infected liver and release into the blood. We developed a slowly dividing liver-derived cell line which multiplies infectious viral particles upon inoculation with patient- or cell culture-derived HBV. This new infection model can improve therapy by measuring, in advance, the sensitivity of a patient’s HBV strain to specific antiviral drugs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Cell culture system that mimicks complete HBV life cycle from entry to egress. </LI> <LI> Efficient <I>in vitro</I> infection with crude HBV patient sera. </LI> <LI> Up to 50- and 1,300-fold net amplification of patient- and cell culture-derived input HBV in the supernatant. </LI> <LI> Polyethylene glycol-independent HBV spread to adjacent cells, forming infected cell clusters. </LI> <LI> Evaluation of patient- and cell culture-derived HBV amplification w/wo antivirals over 8 weeks. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

<i>K</i>-mixing in the doubly mid-shell nuclide ¹⁷⁰Dy and the role of vibrational degeneracy

S&ouml,derstr&ouml,m, P.-A.,Walker, P.M.,Wu, J.,Liu, H.L.,Regan, P.H.,Watanabe, H.,Doornenbal, P.,Korkulu, Z.,Lee, P.,Liu, J.J.,Lorusso, G.,Nishimura, S.,Phong, V.H.,Sumikama, T.,Xu, F.R.,Yagi, A.,Zha North-Holland Pub. Co 2016 Physics letters. Section B Vol.762 No.-

<P><B>Abstract</B></P> <P>A detailed study of the structure of the doubly mid-shell nucleus Dy 104 1 66 170 has been carried out, following isomeric and <I>β</I> decay. We have measured the yrast band up to the spin-parity <SUP> J π </SUP> = <SUP> 6 + </SUP> state, the K = 2 <I>γ</I>-vibration band up to the <SUP> 5 + </SUP> state, a low-lying negative-parity band based on a <SUP> 2 − </SUP> state that could be a candidate for the lowest energy octupole vibration state within this nucleus, and a candidate for the <SUP> K π </SUP> = <SUP> 6 + </SUP> two quasi-particle isomer. This state was determined to have an excitation energy of 1643.91(23) keV and a half life of 0.99(4) μs, with a reduced hindrance for its decay to the ground-state band an order of magnitude lower than predicted by <SUB> N p </SUB> <SUB> N n </SUB> systematics. This is interpreted as being due to <I>γ</I>-vibrational mixing from a near degeneracy of the isomer and the <SUP> 6 + </SUP> state of the <I>γ</I> band. Furthermore, the parent nucleus <SUP>170</SUP>Tb has been determined to have a half-life of 0.91 ( − 13 + 18 ) s with a possible spin-parity of <SUP> 2 − </SUP> .</P>

Reorganization of Southern Ocean Plankton Ecosystem at the Onset of Antarctic Glaciation

Houben, Alexander J. P.,Bijl, Peter K.,Pross, J&ouml,rg,Bohaty, Steven M.,Passchier, Sandra,Stickley, Catherine E.,R&ouml,hl, Ursula,Sugisaki, Saiko,Tauxe, Lisa,van de Flierdt, Tina,Olney, Matthew,San American Association for the Advancement of Scienc 2013 Science Vol.340 No.6130

<P><B>Southern Change</B></P><P>Antarctica has been mostly covered by ice since the inception of large-scale continental glaciation during the Oligocene, which profoundly altered the isotopic and mineralogical records of the sediments surrounding the continent. <B>Houben <I>et al.</I></B> (p. 341) found records of the corresponding living systems in the fossil marine dinoflagellate cysts, which revealed that a microplankton ecosystem, similar to the one that exists today, appeared simultaneously with the first major Antarctic glaciation approximately 34 million years ago.</P>

Novel graphene/Sn and graphene/SnO_x hybrid nanostructures: Induced superconductivity and band gaps revealed by scanning probe measurements

Pá,linká,s, Andrá,s,Molná,r, Gy&ouml,rgy,Magda, Gá,bor Zsolt,Hwang, Chanyong,Tapasztó,, Levente,Samuely, Peter,Szabó,, Pavol,Osvá,th, Zoltá,n Elsevier 2017 Carbon Vol.124 No.-

<P>The development of functional composite nanomaterials based on graphene and metal nanoparticles (NPs) is currently the subject of intense research interest. In this study we report the preparation of novel type of graphene/Sn and graphene/SnOx (1 <= x <= 2) hybrid nanostructures and their investigation by scanning probe methods. First, we prepare Sn NPs by evaporating 7-8 nm tin on highly oriented pyrolytic graphite substrates. Graphene/Sn nanostructures are obtained by transferring graphene on top of the tin NPs immediately after evaporation. We show by scanning tunnelling microscopy (STM) and spectroscopy (STS) that tin NPs reduce significantly the environmental p-type doping of graphene. Furthermore, we demonstrate by low-temperature STM and STS measurements that superconductivity is induced in graphene, either directly supported by Sn NPs or suspended between them. Additionally, we prepare SnOx NPs by annealing the evaporated tin at 500 degrees C. STS measurements performed on hybrid graphene/SnOx nanostructures reveal the electronic band gap of SnOx NPs. The results can open new avenues for the fabrication of novel hybrid superconducting nanomaterials with designed structures and morphologies. (C) 2017 Elsevier Ltd. All rights reserved.</P>

Integrated Navigation and Sense & Avoid Systems for Micro Aerial Vehicles

P,V&ouml,rsmann,S,Winkler,J,-B,Park 한국항해항만학회 2006 한국항해항만학회 학술대회논문집 Vol.1 No.-

The paper deals with integrated navigation and sense & avoid systems for small unmanned aerial vehicles (UAV). First an introduction to the current UAV activities of the institute is given. It is followed by an overview about the integrated navigation system developed for small UAVs. The system is based on a tightly-coupled GPS/INS architec-ture. But instead of using delta-ranges, time-differenced carrier phases are used to aid the INS. Finally, results from navigation filter validation in flight tests are presented. After that an overview about sense and avoid strategies for application in small unmanned aircraft is given. From this a guideline for developing such a system for the institute’s UAVs is presented.

SQUIDs in biomagnetism: a roadmap towards improved healthcare

K&ouml,rber, Rainer,Storm, Jan-Hendrik,Seton, Hugh,Mä,kelä,, Jyrki P,Paetau, Ritva,Parkkonen, Lauri,Pfeiffer, Christoph,Riaz, Bushra,Schneiderman, Justin F,Dong, Hui,Hwang, Seong-min,You, Lixi IOP 2016 Superconductor science & technology Vol.29 No.11

<P>Globally, the demand for improved health care delivery while managing escalating costs is a major challenge. Measuring the biomagnetic fields that emanate from the human brain already impacts the treatment of epilepsy, brain tumours and other brain disorders. This roadmap explores how superconducting technologies are poised to impact health care. Biomagnetism is the study of magnetic fields of biological origin. Biomagnetic fields are typically very weak, often in the femtotesla range, making their measurement challenging. The earliest <I>in vivo</I> human measurements were made with room-temperature coils. In 1963, Baule and McFee (1963 <I>Am</I>. <I>Heart J</I>. <A HREF='http://dx.doi.org/10.1016/0002-8703(63)90075-9'> <B>55</B> 95−6</A>) reported the magnetic field produced by electric currents in the heart (‘magnetocardiography’), and in 1968, Cohen (1968 <I>Science</I> <A HREF='http://dx.doi.org/10.1126/science.161.3843.784'> <B>161</B> 784−6</A>) described the magnetic field generated by alpha-rhythm currents in the brain (‘magnetoencephalography’). Subsequently, in 1970, Cohen <I>et al</I> (1970 <I>Appl. Phys. Lett.</I> <A HREF='http://dx.doi.org/10.1063/1.1653195'> <B>16</B> 278–80</A>) reported the recording of a magnetocardiogram using a Superconducting QUantum Interference Device (SQUID). Just two years later, in 1972, Cohen (1972 <I>Science</I> <A HREF='http://dx.doi.org/10.1126/science.175.4022.664'> <B>175</B> 664–6</A>) described the use of a SQUID in magnetoencephalography. These last two papers set the scene for applications of SQUIDs in biomagnetism, the subject of this roadmap.</P> <P>The SQUID is a combination of two fundamental properties of superconductors. The first is flux quantization—the fact that the magnetic flux Φ in a closed superconducting loop is quantized in units of the magnetic flux quantum, Φ<SUB>0</SUB> ≡ <I>h</I>/2<I>e</I>, ≈ 2.07 × 10<SUP>−15</SUP> Tm<SUP>2</SUP> (Deaver and Fairbank 1961 <I>Phys. Rev. Lett.</I> <A HREF='http://dx.doi.org/10.1103/PhysRevLett.7.43'> <B>7</B> 43–6</A>, Doll R and Näbauer M 1961 <I>Phys. Rev. Lett.</I> <A HREF='http://dx.doi.org/10.1103/PhysRevLett.7.51'> <B>7</B> 51–2</A>). Here, <I>h</I> is the Planck constant and <I>e</I> the elementary charge. The second property is the Josephson effect, predicted in 1962 by Josephson (1962 <I>Phys. Lett.</I> <A HREF='http://dx.doi.org/10.1016/0031-9163(62)91369-0'> <B>1</B> 251–3</A>) and observed by Anderson and Rowell (1963 <I>Phys. Rev. Lett.</I> <A HREF='http://dx.doi.org/10.1103/PhysRevLett.10.230'> <B>10</B> 230–2</A>) in 1963. The Josephson junction consists of two weakly coupled superconductors separated by a tunnel barrier or other weak link. A tiny electric current is able to flow between the superconductors as a supercurrent, without developing a voltage across them. At currents above the ‘critical current’ (maximum supercurrent), however, a voltage is developed. In 1964, Jaklevic <I>et al</I> (1964 <I>Phys. Rev. Lett.</I> <A HREF='http://dx.doi.org/10.1103/PhysRevLett.12.159'> <B>12</B> 159–60</A>) observed quantum interference between two Josephson junctions connected in series on a superconducting loop, giving birth to the dc SQUID. The essential property of the SQUID is that a steady increase in the magnetic flux threading the loop causes the critical current to oscillate with a period of one flux quantum. In today’s SQUIDs, using conventional semiconductor readout electronics, one can typically detect a change in Φ corresponding to 10<SUP>−6</SUP> Φ<SUB>0</SUB> in one second. Although early practical SQUIDs were usually made from bulk superconductors, for example, niobium or Pb-Sn solder blobs, today’s devices are invariably made from thin superconducting films patterned with photolithography or even electron lithography. An extensive descri

An in vitro comparison of possibly bioactive titanium implant surfaces

G&ouml,ransson, A.,Arvidsson, A.,Currie, F.,Franke-Stenport, V.,Kjellin, P.,Mustafa, K.,Sul, Y. T.,Wennerberg, A. Wiley Subscription Services, Inc., A Wiley Company 2009 Journal of biomedical materials research. Part A Vol.a88 No.4

<P>The aim of the study was to compare Ca and P formation (CaP) and subsequent bone cell response of a blasted and four different possibly bioactive commercially pure (cp) titanium surfaces; 1. Fluoride etched (Fluoride), 2. Alkali-heat treated (AH), 3. Magnesium ion incorporated anodized (TiMgO), and 4. Nano HA coated and heat treated (nano HA) in vitro. Furthermore, to evaluate the significance of the SBF formed CaP coat on bone cell response. The surfaces were characterized by Optical Interferometry, Scanning Electron Microscopy (SEM) and X-ray Photoelectron Spectroscopy (XPS). CaP formation was evaluated after 12, 24 and 72 h in simulated body fluid (SBF). Primary human mandibular osteoblast-like cells were cultured on the various surfaces subjected to SBF for 72 h. Cellular attachment, differentiation (osteocalcin) and protein production (TGF-β<SUB>1</SUB>) was evaluated after 3 h and 10 days respectively. Despite different morphological appearances, the roughness of the differently modified surfaces was similar. The possibly bioactive surfaces gave rise to an earlier CaP formation than the blasted surface, however, after 72 h the blasted surface demonstrated increased CaP formation compared to the possibly bioactive surfaces. Subsequent bone cell attachment was correlated to neither surface roughness nor the amount of formed CaP after SBF treatment. In contrast, osteocalcin and TGF-β<SUB>1</SUB> production were largely correlated to the amount of CaP formed on the surfaces. However, bone response (cell attachment, osteocalcin and TGF-F production) on the blasted controls were similar or increased compared to the SBF treated fluoridated, AH and TiMgO surface. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009</P>

Two-hole structure outsideNi78: Existence of aμsisomer ofCo76andβdecay intoNi76

S&ouml,derstr&ouml,m, P.-A.,Nishimura, S.,Xu, Z. Y.,Sieja, K.,Werner, V.,Doornenbal, P.,Lorusso, G.,Browne, F.,Gey, G.,Jung, H. S.,Sumikama, T.,Taprogge, J.,Vajta, Zs.,Watanabe, H.,Wu, J.,Baba, H.,Dom American Physical Society 2015 PHYSICAL REVIEW C - Vol.92 No.5

High tolerance of hepatitis B virus to thermal disinfection

K&ouml,nig, Alexander,Than, Thoa Thi,Todt, Daniel,Yoon, Seung Kew,Steinmann, Jochen,Steinmann, Eike,Windisch, Marc P. Elsevier 2019 Journal of hepatology Vol.71 No.6

Promoter methylation of the Wnt/β‐catenin signaling antagonist <i>Dkk‐3</i> is associated with poor survival in gastric cancer

Yu, Jun,Tao, Qian,Cheng, Yuen Y.,Lee, Kwan Y.,Ng, Simon S. M.,Cheung, Kin F.,Tian, Linwei,Rha, Sun Y.,Neumann, Ulf,R&ouml,cken, Christoph,Ebert, Matthias P. A.,Chan, Francis K. L.,Sung, Joseph J. Y. Wiley Subscription Services, Inc., A Wiley Company 2009 Cancer Vol.115 No.1

<P><B>Abstract</B></P><P><B>BACKGROUND:</B></P><P>Abnormal activation of the Wnt/β‐catenin signaling pathway is common and critical in the pathogenesis of digestive cancers. In this study, the authors investigated the promoter methylation of the dickkopf homolog 3 gene <I>Dkk‐3</I> in these cancers and its prognostic significance in gastric cancer.</P><P><B>METHODS:</B></P><P><I>Dkk‐3</I> methylation was assessed in 173 patients with gastric cancers (including 104 patients who were followed for up to 4090 days) and in 128 patients with colorectal cancer. Cell growth was evaluated by using a colony‐formation assay. For survival analyses, the authors used Kaplan‐Meier plots, the log‐rank test, and Cox proportional regression.</P><P><B>RESULTS:</B></P><P><I>Dkk‐3</I> was silenced or down‐regulated in 12 of 17 gastric cancer cell lines (70.6%) and in 3 of 9 colon cancer cell lines (33.3%). The loss of gene expression was associated with promoter methylation, which could be restored by demethylating agents. Ectopic expression of <I>Dkk‐3</I> suppressed colony formation. Moreover, methylation of <I>Dkk‐3</I> was detected in 117 of 173 primary gastric tumors (67.6%) and in 67 of 128 colorectal tumors (52.3%). The clinical significance and the prognostic value of <I>Dkk‐3</I> methylation also were examined in 104 gastric cancers and in 84 colorectal cancers. Multivariate analysis indicated that <I>Dkk‐3</I> methylation was associated significantly and independently with poor disease survival (relative risk, 2.534; 95% confidence interval, 1.54–4.17; <I>P</I> = .002) in gastric cancer, but not in colorectal cancer. Kaplan‐Meier survival curves revealed that patients who had <I>Dkk‐3</I> methylated gastric cancers had a significantly shorter survival (median, 0.76 years) compared with patients who did not have <I>Dkk‐3</I> methylation (median, 2.68 years; <I>P</I> < .0001; log‐rank test).</P><P><B>CONCLUSIONS:</B></P><P>Epigenetic silencing of the <I>Dkk‐3</I> gene by promoter methylation was a common event in gastric cancer and was associated with a poor outcome in such patients. Cancer 2009. © 2008 American Cancer Society.</P>