bluuebunny/crossref_metadata_2025 huggingface dataset:
Should the standard dimethyl sulfoxide concentration be reduced? Results of a European Group for Blood and Marrow Transplantation prospective noninterventional study on usage and side effects of dimethyl sulfoxide
Authors: Morris Curly , de Wreede Liesbeth , Scholten Marijke , Brand Ronald , van Biezen Anja , Sureda Anna , Dickmeiss Ebbe , Trneny Marek , Apperley Jane , Chiusolo Patrizia , van Imhoff Gustaaf W. , Lenhoff Stig , Martinelli Giovanni , Hentrich Marcus , Pabst Thomas , Onida Francesco , Quinn Michael , Kroger Nicolaus , de Witte Theo , Ruutu Tapani ,   Chronic Malignancies and Lymphoma Working Parties of the EBMT
Published: June 2014

Background

Dimethyl sulfoxide (DMSO) is essential for the preservation of liquid nitrogen–frozen stem cells, but is associated with toxicity in the transplant recipient.

Study Design and Methods

In this prospective noninterventional study, we describe the use of DMSO in 64 European Blood and Marrow Transplant Group centers undertaking autologous transplantation on patients with myeloma and lymphoma and analyze side effects after return of DMSO‐preserved stem cells.

Results

While the majority of centers continue to use 10% DMSO, a significant proportion either use lower concentrations, mostly 5 or 7.5%, or wash cells before infusion (some for selected patients only). In contrast, the median dose of DMSO given (20 mL) was much less than the upper limit set by the same institutions (70 mL). In an accompanying statistical analysis of side effects noted after return of DMSO‐preserved stem cells, we show that patients in the highest quartile receiving DMSO (mL and mL/kg body weight) had significantly more side effects attributed to DMSO, although this effect was not observed if DMSO was calculated as mL/min. Dividing the myeloma and lymphoma patients each into two equal groups by age we were able to confirm this result in all but young myeloma patients in whom an inversion of the odds ratio was seen, possibly related to the higher dose of melphalan received by young myeloma patients.

Conclusion

We suggest better standardization of preservation method with reduced DMSO concentration and attention to the dose of DMSO received by patients could help reduce the toxicity and morbidity of the transplant procedure.
Novel swine model of transfusion‐related acute lung injury
Authors: Okazaki Hitoshi , Ishikawa Osamu , Iijima Takehiko , Kohira Takahiro , Teranishi Mai , Kawasaki Shin , Saito Akira , Mikami Yu , Sugiura Asuka , Hashimoto Shiho , Shimada Eiko , Uchikawa Makoto , Matsuhashi Mika , Tsuno Nelson H. , Tanaka Minoru , Kiyokawa Nobutaka , Fujimoto Junichiro , Nagase Takahide , Tadokoro Kenji , Takahashi Koki
Published: June 2014

Background

Transfusion‐related acute lung injury (TRALI) is a life‐threatening complication of blood transfusion. Antibodies against human leukocyte antigens in donors' plasma are the major causes of TRALI. Several animal models of TRALI have been developed, and the mechanism underlying TRALI development has been extensively investigated using rodent models. Although sheep models of nonimmune TRALI have been developed, large‐animal models of antibody‐mediated TRALI are not yet available.

Study Design and Methods

To develop a swine model of TRALI, male Clawn strain miniature pigs were used. A monoclonal antibody (MoAb) against swine leukocyte antigens (SLAs) Class I (4G8, 0.3 or 1.0 mg/kg body weight [BW]) and a control antibody (1.0 mg/kg BW) were injected into the peripheral vein after priming with or without 1 μg/kg BW lipopolysaccharide (LPS; n = 3 each). Lung injury was assessed using PaO2/FiO2 (P/F) ratio and by chest X‐ray imaging. Histopathologic analysis was also conducted.

Results

Lung injury could be induced by injecting 4G8 at an amount of 1.0 mg/kg BW, after LPS. The P/F ratio 90 minutes after the administration of 4G8 significantly decreased (p < 0.05). Bilateral infiltration was shown in chest X‐ray imaging. Lung injury was confirmed by histopathologic analysis.

Conclusion

Lung injury in pigs was successfully induced by anti‐SLA MoAb. Priming with LPS is a prerequisite for inducing lung injury and the amount of the antibody is a critical condition.
Biosimilar granulocyte–colony‐stimulating factor for healthy donor stem cell mobilization: need we be afraid?
Authors: Bonig Halvard , Becker Petra S. , Schwebig Arnd , Turner Matthew
Published: June 2014

Biosimilars are approved biologics with comparable quality, safety, and efficacy to a reference product. Unlike generics, which are chemically manufactured copies of small‐molecule drugs with relatively simple chemical structures, the biosimilar designation is applied to drugs that are produced by living organisms, implying much more difficult to control manufacturing and purification procedures. To account for these complexities, the European Medicines Agency (EMA), the US Food and Drug Administration, the Australian Therapeutic Goods Administration, and other regulatory authorities have devised and implemented specific, markedly more demanding pathways for the evaluation and approval of biosimilars. To date, several biosimilars have been approved, including versions of somatropin, erythropoietin, and granulocyte–colony‐stimulating factor (G‐CSF), and several biosimilar monoclonal antibodies are currently in development. The reference G‐CSF product (Neupogen, Amgen) has been used for many years for prevention and treatment of neutropenia and also for mobilization of peripheral blood stem cells (PBSCs). However, concerns have been raised about the safety and efficacy of biosimilar G‐CSF during PBSC mobilization procedures, especially in healthy donors. This article reviews the available evidence on the use of biosimilar G‐CSF in this setting. Aggregate clinical evidence supports the assessment by the EMA of biosimilar and originator G‐CSF as highly biologically similar, with respect to desired and undesired effects.
Onset of Wiggling in a Microscopic Patterned Structure Induced by Intrinsic Stress During the Dry Etching Process
Authors: Tanaka Hiro , Hidaka Takahiro , Izumi Satoshi , Sakai Shinsuke
Published: June 2014

In semiconductor devices, fine patterning can cause structural instability because of intrinsic compressive stress. We studied one such instability phenomenon, out-of-plane wiggling of a patterned structure with mask–dielectric ridges, to improve the yield of these highly miniaturized devices. Our simple continuum approach uses dimensionless parameters to control the bifurcation threshold of ridge wiggling. Coupled with modeling the etching process, our approach revealed the onset of buckling, agreeing well with experimental data. To study the influence of the ridge width and the elastic substrate on buckling stress and deformation, we performed numerical analyses using a finite element method (FEM).
Psychophysical auditory filter estimates reveal sharper cochlear tuning in musicians
Authors: Bidelman Gavin M. , Schug Jonathan M. , Jennings Skyler G. , Bhagat Shaum P.
Published: June 2014

Musicianship confers enhancements to hearing at nearly all levels of the auditory system from periphery to percept. Musicians' superior psychophysical abilities are particularly evident in spectral discrimination and noise-degraded listening tasks, achieving higher perceptual sensitivity than their nonmusician peers. Greater spectral acuity implies that musicianship may increase auditory filter selectivity. This hypothesis was directly tested by measuring both forward- and simultaneous-masked psychophysical tuning curves. Sharper filter tuning (i.e., higher Q10) was observed in musicians compared to nonmusicians. Findings suggest musicians' pervasive listening benefits may be facilitated, in part, by superior spectral processing/decomposition as early as the auditory periphery.
“I had no desire to be having this battle with this faceless man on the soccer field anymore”: Exploring the Ethics of Sporting Retirement
Authors: Crocket Hamish
Published: June 2014

In this article, I examine the relatively youthful sporting retirement of four athletes from popular Western team sports, namely soccer, ice hockey and rugby union. Drawing on studies of sporting transitions and retirement (e.g., Denison, 1997; Douglas & Carless, 2009; Sparkes, 1998; Sparkes & Smith, 2002; Stambulova, Alfermann, Statler, & Côté, 2009; Wylleman, Alfermann, & Lavallee, 2004) and Foucault’s notions of games of truth and ethical self-creation, I argue that these athletes’ decisions to retire from sport were based on a refusal to accept the subject position proposed to them within their sports and, subsequently, these sporting transitions formed acts of ethical self-creation. My conclusions reveal an as yet under-explored interpretation of sporting retirement in which the process of retirement forms part of an attempt to recreate an ethical self.
Global epistasis makes adaptation predictable despite sequence-level stochasticity
Authors: Kryazhimskiy Sergey , Rice Daniel P. , Jerison Elizabeth R. , Desai Michael M.
Published: June 2014

Clouding evolution's crystal ball

Because of a sort of mutation buffering process, different starting mutations can tend to end up with similar overall affects on an organism's fitness. Kryazhimskiy
et al.
evolved lines of yeast, each originating from distinct single genotypes, under the same selective regimen. A subset of clones from these adapted populations was subjected to fitness assays and sequenced. Populations with lower initial fitness, adapted more rapidly than populations with higher initial fitness, so that in the end the fitness levels were similar.

Science
, this issue p.
1519
Anticipating the next century of wastewater treatment
Authors: van Loosdrecht Mark C. M. , Brdjanovic Damir
Published: June 2014

Advances in activated sludge sewage treatment can improve its energy use and resource recovery
Not-seeing is believing
Authors: Stone Richard
Published: June 2014

“Zero knowledge” tests could let inspectors identify a nuclear weapon without learning the secrets of its design.
Daring to live on the edge
Authors: Pennisi Elizabeth
Published: June 2014

Tracking the house sparrow's spread in Africa yields secrets of these invaders—a taste for novel foods, for example.
Drawing a bead on trade
Authors: Lawler Andrew
Published: June 2014

The humble glass bead has become a tool to trace the migration of ancient goods and peoples around the Indian Ocean.
The gas surge
Authors: Malakoff David
Published: June 2014

Fracking has ignited an energy revolution, with still-uncertain consequences for climate and the environment.
Searching for life in the deep shale
Authors: Pennisi Elizabeth
Published: June 2014

Energy developers and researchers alike want to find out what's living in the Marcellus Shale's deep layers of rock.
Hunting a climate fugitive
Authors: Kintisch Eli
Published: June 2014

Plugging methane leaks in the urban maze could be key to making shale gas climate-friendly.
The bond breaker
Authors: Service Robert
Published: June 2014

By transforming methane, chemist Roy Periana aims to turn natural gas into cheap feedstocks for chemical firms.
Immunogenicity of a Synthetic Vaccine Based on Plasmodium vivax Duffy Binding Protein Region II
Authors: Ntumngia Francis B. , Barnes Samantha J. , McHenry Amy M. , George Miriam T. , Schloegel Jesse , Adams John H.
Published: June 2014

Molecules that play a role inPlasmodiummerozoite invasion of host red blood cells represent attractive targets for blood-stage vaccine development against malaria. InPlasmodium vivax, merozoite invasion of reticulocytes is mediated by the Duffy binding protein (DBP), which interacts with its cognate receptor, the Duffy antigen receptor for chemokines, on the surface of reticulocytes. The DBP ligand domain, known as region II (DBPII), contains the critical residues for receptor recognition, making it a prime target for vaccine development against blood-stage vivax malaria. In natural infections, DBP is weakly immunogenic and DBPII allelic variation is associated with strain-specific immunity, which may compromise vaccine efficacy. In a previous study, a synthetic vaccine termed DEKnull that lacked an immunodominant variant epitope in DBPII induced functional antibodies to shared neutralizing epitopes on the native Sal1 allele. Anti-DEKnull antibody titers were lower than anti-Sal1 titers but produced more consistent, strain-transcending anti-DBPII inhibitory responses. In this study, we further characterized the immunogenicity of DEKnull, finding that immunization with recombinant DEKnull produced an immune response comparable to that obtained with native recombinant DBP alleles. Further investigation of DEKnull is necessary to enhance its immunogenicity and broaden its specificity.
Identification of Occult Fusobacterium nucleatum Central Nervous System Infection by Use of PCR-Electrospray Ionization Mass Spectrometry
Authors: Nagalingam Sudha , Lisgaris Michelle , Rodriguez Benigno , Jacobs Michael R. , Lederman Michael , Salata Robert A , Hujer Andrea M. , Muehlenbachs Atis , DeLeon-Carnes Marlene , Farrell John J. , Sampath Rangarajan , Bonomo Robert A.
Published: June 2014

Anaerobic bacteria are often difficult to detect, especially after the initiation of antibiotics. We describe the application of PCR-electrospray ionization mass spectrometry (PCR/ESI-MS) using a sample of cerebrospinal fluid to identify an anaerobic Gram-negative bacillus,
Fusobacterium nucleatum
, in a patient with “culture-negative” meningitis and cerebral abscesses.
Subcutaneous Phaeohyphomycotic Nodule Due to Phialemoniopsis hongkongensis sp. nov
Authors: Tsang Chi-Ching , Chan Jasper F. W. , Ip Philip P. C. , Ngan Antonio H. Y. , Chen Jonathan H. K. , Lau Susanna K. P. , Woo Patrick C. Y.
Published: June 2014

Phialemoniopsis
species are ubiquitous dematiaceous molds associated with a wide variety of superficial and systemic infections in human. In this study, we isolated a mold from the forearm nodule biopsy specimen from a patient with underlying liver cirrhosis, ankylosing spondylosis, and tuberculosis. He was treated with itraconazole, but unfortunately, he succumbed as a result of disseminated tuberculosis with multiorgan failure. The histology results of the skin biopsy showed necrotizing granulomas in which numerous fungal elements were found. On Sabouraud dextrose agar, the fungal isolate grew as white-to-cream and smooth-to-velvety colonies. Microscopically, oval-to-cylindrical conidia were observed from abundant adelophialides, which possessed barely visible parallel collarettes but no basal septa. The azole drugs voriconazole, itraconazole, and posaconazole, as well as amphotericin B, showed high activities against this fungus. Internal transcribed spacer, 28S nuclear ribosomal DNA (nrDNA), and β-actin and β-tubulin gene sequencing showed that this fungus is most closely related to but distinct from
Phialemonium curvata
. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and hierarchical cluster analysis showed that the MALDI-TOF MS spectrum of this fungus is most closely related to that of
Phialemonium pluriloculosa
. We propose a new species,
Phialemoniopsis hongkongensis
sp. nov., to describe this fungus.
The Linker Region of NS3 Plays a Critical Role in the Replication and Infectivity of Hepatitis C Virus
Authors: Kohlway Andrew , Pirakitikulr Nathan , Ding Steve C. , Yang Feng , Luo Dahai , Lindenbach Brett D. , Pyle Anna M.
Published: June 2014

Hepatitis C virus (HCV) NS3-4A is required for viral replication and assembly. We establish that virus assembly is sensitive to mutations in the linker region between the helicase and protease domains of NS3-4A. However, we find that the protease cleavage, RNA binding, and unwinding rates of NS3 are minimally affected
in vitro
. Thus, we conclude that the NS3 linker is critical for mediating protein-protein interactions and dynamic control rather than for modulating the enzymatic functions of NS3-4A.
A Novel Partitivirus That Confers Hypovirulence on Plant Pathogenic Fungi
Authors: Xiao Xueqiong , Cheng Jiasen , Tang Jinghua , Fu Yanping , Jiang Daohong , Baker Timothy S. , Ghabrial Said A. , Xie Jiatao
Published: June 2014

Members of the family
Partitiviridae
have bisegmented double-stranded RNA (dsRNA) genomes and are not generally known to cause obvious symptoms in their natural hosts. An unusual partitivirus,
Sclerotinia sclerotiorum
partitivirus 1 (SsPV1/WF-1), conferred hypovirulence on its natural plant-pathogenic fungal host,
Sclerotinia sclerotiorum
strain WF-1. Cellular organelles, including mitochondria, were severely damaged. Hypovirulence and associated traits of strain WF-1 and SsPV1/WF-1 were readily cotransmitted horizontally via hyphal contact to different vegetative compatibility groups of
S. sclerotiorum
and interspecifically to
Sclerotinia nivalis
and
Sclerotinia minor
.
S. sclerotiorum
strain 1980 transfected with purified SsPV1/WF-1 virions also exhibited hypovirulence and associated traits similar to those of strain WF-1. Moreover, introduction of purified SsPV1/WF-1 virions into strain KY-1 of
Botrytis cinerea
also resulted in reductions in virulence and mycelial growth and, unexpectedly, enhanced conidial production. However, virus infection suppressed hyphal growth of most germinating conidia of
B. cinerea
and was eventually lethal to infected hyphae, since very few new colonies could develop following germ tube formation. Taken together, our results support the conclusion that SsPV1/WF-1 causes hypovirulence in
Sclerotinia
spp. and
B. cinerea
. Cryo-EM (cryo-electron microscopy) reconstruction of the SsPV1 particle shows that it has a distinct structure with similarity to the closely related partitiviruses
Fusarium poae
virus 1 and
Penicillium stoloniferum
virus F. These findings provide new insights into partitivirus biological activities and clues about molecular interactions between partitiviruses and their hosts.

IMPORTANCE
Members of the
Partitiviridae
are believed to occur commonly in their phytopathogenic fungal and plant hosts. However, most partitiviruses examined so far appear to be associated with latent infections. Here we report a partitivirus, SsPV1/WF-1, that was isolated from a hypovirulent strain of
Sclerotinia sclerotiorum
and describe its biological and molecular features. We have demonstrated that SsPV1 confers hypovirulence. Furthermore, SsPV1 can infect and cause hypovirulence in
Botrytis cinerea
. Our study also suggests that SsPV1 has a vigorous ability to proliferate and spread via hyphal contact. SsPV1 can overcome vegetative incompatibility barriers and can be transmitted horizontally among different vegetative compatibility groups of
S. sclerotiorum
, even interspecifically. Cryo-EM reconstruction of SsPV1 shows that it has a distinct structure with similarity to closely related partitiviruses. Our studies exploit a novel system, SsPV1 and its hosts, which can provide the means to explore the mechanisms by which partitiviruses interact with their hosts.
Virus-Induced Tubule: a Vehicle for Rapid Spread of Virions through Basal Lamina from Midgut Epithelium in the Insect Vector
Authors: Jia Dongsheng , Mao Qianzhuo , Chen Hongyan , Wang Aiming , Liu Yuyan , Wang Haitao , Xie Lianhui , Wei Taiyun
Published: June 2014

The plant reoviruses, plant rhabdoviruses, tospoviruses, and tenuiviruses are transmitted by insect vectors in a persistent propagative manner. These viruses induce the formation of viral inclusions to facilitate viral propagation in insect vectors. The intestines of insect vectors are formed by epithelial cells that lie on the noncellular basal lamina surrounded by visceral muscle tissue. Here, we demonstrate that a recently identified plant reovirus, southern rice black-streaked dwarf virus (SRBSDV), exploits virus-containing tubules composed of virus-encoded nonstructural protein P7-1 to directly cross the basal lamina from the initially infected epithelium toward visceral muscle tissues in the intestine of its vector, the white-backed planthopper (
Sogatella furcifera
). Furthermore, such tubules spread along visceral muscle tissues through a direct interaction of P7-1 and actin. The destruction of tubule assembly by RNA interference with synthesized double-stranded RNA targeting the P7-1 gene inhibited viral spread in the insect vector
in vitro
and
in vivo
. All these results show for the first time that a virus employs virus-induced tubule as a vehicle for viral spread from the initially infected midgut epithelium through the basal lamina, facilitating the rapid dissemination of virus from the intestine of the insect vector.

IMPORTANCE
Numerous plant viruses are transmitted in a persistent manner by sap-sucking insects, including thrips, aphids, planthoppers, and leafhoppers. These viruses, ingested by the insects, establish their primary infection in the intestinal epithelium of the insect vector. Subsequently, the invading virus manages to transverse the basal lamina, a noncellular layer lining the intestine, a barrier that may theoretically hinder viral spread. The mechanism by which plant viruses cross the basal lamina is unknown. Here, we report that a plant virus has evolved to exploit virus-induced tubules to pass through the basal lamina from the initially infected midgut epithelium of the insect vector, thus revealing the previously undescribed pathway adapted by the virus for rapid dissemination of virions from the intestine of the insect vector.
A Novel Life Cycle Modeling System for Ebola Virus Shows a Genome Length-Dependent Role of VP24 in Virus Infectivity
Authors: Watt Ari , Moukambi Felicien , Banadyga Logan , Groseth Allison , Callison Julie , Herwig Astrid , Ebihara Hideki , Feldmann Heinz , Hoenen Thomas
Published: June 2014

Work with infectious Ebola viruses is restricted to biosafety level 4 (BSL4) laboratories, presenting a significant barrier for studying these viruses. Life cycle modeling systems, including minigenome systems and transcription- and replication-competent virus-like particle (trVLP) systems, allow modeling of the virus life cycle under BSL2 conditions; however, all current systems model only certain aspects of the virus life cycle, rely on plasmid-based viral protein expression, and have been used to model only single infectious cycles. We have developed a novel life cycle modeling system allowing continuous passaging of infectious trVLPs containing a tetracistronic minigenome that encodes a reporter and the viral proteins VP40, VP24, and GP
1,2
. This system is ideally suited for studying morphogenesis, budding, and entry, in addition to genome replication and transcription. Importantly, the specific infectivity of trVLPs in this system was ∼500-fold higher than that in previous systems. Using this system for functional studies of VP24, we showed that, contrary to previous reports, VP24 only very modestly inhibits genome replication and transcription when expressed in a regulated fashion, which we confirmed using infectious Ebola viruses. Interestingly, we also discovered a genome length-dependent effect of VP24 on particle infectivity, which was previously undetected due to the short length of monocistronic minigenomes and which is due at least partially to a previously unknown function of VP24 in RNA packaging. Based on our findings, we propose a model for the function of VP24 that reconciles all currently available data regarding the role of VP24 in nucleocapsid assembly as well as genome replication and transcription.

IMPORTANCE
Ebola viruses cause severe hemorrhagic fevers in humans, with no countermeasures currently being available, and must be studied in maximum-containment laboratories. Only a few of these laboratories exist worldwide, limiting our ability to study Ebola viruses and develop countermeasures. Here we report the development of a novel reverse genetics-based system that allows the study of Ebola viruses without maximum-containment laboratories. We used this system to investigate the Ebola virus protein VP24, showing that, contrary to previous reports, it only modestly inhibits virus genome replication and transcription but is important for packaging of genomes into virus particles, which constitutes a previously unknown function of VP24 and a potential antiviral target. We further propose a comprehensive model for the function of VP24 in nucleocapsid assembly. Importantly, on the basis of this approach, it should easily be possible to develop similar experimental systems for other viruses that are currently restricted to maximum-containment laboratories.
High-Throughput Identification of Loss-of-Function Mutations for Anti-Interferon Activity in the Influenza A Virus NS Segment
Authors: Wu Nicholas C. , Young Arthur P. , Al-Mawsawi Laith Q. , Olson C. Anders , Feng Jun , Qi Hangfei , Luan Harding H. , Li Xinmin , Wu Ting-Ting , Sun Ren
Published: June 2014

Viral proteins often display several functions which require multiple assays to dissect their genetic basis. Here, we describe a systematic approach to screen for loss-of-function mutations that confer a fitness disadvantage under a specified growth condition. Our methodology was achieved by genetically monitoring a mutant library under two growth conditions, with and without interferon, by deep sequencing. We employed a molecular tagging technique to distinguish true mutations from sequencing error. This approach enabled us to identify mutations that were negatively selected against, in addition to those that were positively selected for. Using this technique, we identified loss-of-function mutations in the influenza A virus NS segment that were sensitive to type I interferon in a high-throughput fashion. Mechanistic characterization further showed that a single substitution, D92Y, resulted in the inability of NS to inhibit RIG-I ubiquitination. The approach described in this study can be applied under any specified condition for any virus that can be genetically manipulated.

IMPORTANCE
Traditional genetics focuses on a single genotype-phenotype relationship, whereas high-throughput genetics permits phenotypic characterization of numerous mutants in parallel. High-throughput genetics often involves monitoring of a mutant library with deep sequencing. However, deep sequencing suffers from a high error rate (∼0.1 to 1%), which is usually higher than the occurrence frequency for individual point mutations within a mutant library. Therefore, only mutations that confer a fitness advantage can be identified with confidence due to an enrichment in the occurrence frequency. In contrast, it is impossible to identify deleterious mutations using most next-generation sequencing techniques. In this study, we have applied a molecular tagging technique to distinguish true mutations from sequencing errors. It enabled us to identify mutations that underwent negative selection, in addition to mutations that experienced positive selection. This study provides a proof of concept by screening for loss-of-function mutations on the influenza A virus NS segment that are involved in its anti-interferon activity.
Fine Mapping of Murine Antibody Responses to Immunization with a Novel Soluble Form of Hepatitis C Virus Envelope Glycoprotein Complex
Authors: Ruwona Tinashe B. , Giang Erick , Nieusma Travis , Law Mansun
Published: June 2014

The hepatitis C virus (HCV) envelope glycoprotein E1E2 complex is a candidate vaccine antigen. Previous immunization studies of E1E2 have yielded various results on its ability to induce virus-neutralizing antibodies in animal models and humans. The murine model has become a vital tool for HCV research owing to the development of humanized mice susceptible to HCV infection. In this study, we investigated the antibody responses of mice immunized with E1E2 and a novel soluble form of E1E2 (sE1E2) by a DNA prime and protein boost strategy. The results showed that sE1E2 elicited higher antibody titers and a greater breadth of reactivity than the wild-type cell-associated E1E2. However, immune sera elicited by either immunogen were only weakly neutralizing. In order to understand the contrasting results of binding and serum neutralizing activities, epitopes targeted by the polyclonal antibody responses were mapped and monoclonal antibodies (MAbs) were generated. The results showed that the majority of serum antibodies were directed to the E1 region 211 to 250 and the E2 regions 421 to 469, 512 to 539, 568 to 609, and 638 to 651, instead of the well-known immunodominant E2 hypervariable region 1 (HVR1). Unexpectedly, in MAb analysis, ∼12% of MAbs isolated were specific to the conserved E2 antigenic site 412 to 423, and 85% of them cross-neutralized multiple HCV isolates. The epitopes recognized by these MAbs are similar but distinct from the previously reported HCV1 and AP33 broadly neutralizing epitopes. In conclusion, E1E2 can prime B cells specific to conserved neutralizing epitopes, but the levels of serum neutralizing antibodies elicited are insufficient for effective virus neutralization. The sE1E2 constructs described in this study can be a useful template for rational antigen engineering.

IMPORTANCE
Hepatitis C virus infects 2 to 3% of the world's population and is a leading cause of liver failures and the need for liver transplantation. The virus envelope glycoprotein complex E1E2 produced by detergent extraction of cells overexpressing the protein was evaluated in a phase I clinical trial but failed to induce neutralizing antibodies in most subjects. In this study, we designed a novel form of E1E2 which is secreted from cells and is soluble and compared it to wild-type E1E2 by DNA immunization of mice. The results showed that this new E1E2 is more immunogenic than wild-type E1E2. Detailed mapping of the antibody responses revealed that antibodies to the conserved E2 antigenic site 412 to 423 were elicited but the serum concentrations were too low to neutralize the virus effectively. This soluble E1E2 provides a new reagent for studying HCV and for rational vaccine design.
Broadly Neutralizing Antibody VRC01 Prevents HIV-1 Transmission from Plasmacytoid Dendritic Cells to CD4 T Lymphocytes
Authors: Su Bin , Lederle Alexandre , Laumond Géraldine , Ducloy Camille , Schmidt Sylvie , Decoville Thomas , Moog Christiane
Published: June 2014

Plasmacytoid dendritic cells (pDC) poorly replicate human immunodeficiency virus type 1 (HIV-1) but efficiently transfer HIV-1 to adjacent CD4 T lymphocytes. We found that coculture with T lymphocytes downregulates SAMHD1 expression, enhances HIV-1 replication, and increases pDC maturation and alpha interferon (IFN-α) secretion. HIV-1 transfer to T lymphocytes is inhibited by broadly neutralizing antibody VRC01 with efficiency similar to that of cell-free infection of T lymphocytes. Interestingly, prevention of HIV-1 transmission by VRC01 retains IFN-α secretion. These results emphasize the multiple functions of VRC01 in protection against HIV-1 acquisition.
Blockade of Type I Interferon (IFN) Production by Retroviral Replicating Vectors and Reduced Tumor Cell Responses to IFN Likely Contribute To Tumor Selectivity
Authors: Lin Amy H. , Burrascano Cindy , Pettersson Par L. , Ibañez Carlos E. , Gruber Harry E. , Jolly Douglas J.
Published: June 2014

We developed a Moloney mouse leukemia virus (MLV)-based retroviral replicating vector (RRV), Toca 511, which has displayed tumor specificity in resected brain tumor material and blood in clinical trials. Here, we investigated the interaction between Toca 511 and human host cells, and we show that RRVs do not induce type I interferon (IFN) responses in cultured human tumor cells or cultured human primary cells. However, exogenous type I IFN inhibited RRV replication in tumor cells and induced IFN-regulated genes, albeit at a lower level than in primary cells. Unexpectedly, RRVs did not induce IFN-α production upon incubation
in vitro
with human plasmacytoid dendritic cells (pDCs), whereas lentivirus vector and heat-treated RRVs did. Coincubation of RRVs with heat-treated RRVs or with lentivirus vector suppressed IFN-α production in pDCs, suggesting that native RRV has a dominant inhibitory effect on type I IFN induction. This effect is sensitive to trypsin treatment. In addition, heat treatment inactivated that activity but exposed an immune-stimulatory activity. The immune-stimulating component is sensitive to deglycosidases, trypsin, and phospholipase C treatment. Experiments with retroviral nonreplicating vectors and virus-like particles demonstrated that the immunosuppressive activity is not associated with the amphotropic envelope or the glyco-Gag protein. In summary, our data provide evidence that RRVs do not directly trigger type I IFN responses in IFN-responsive tumor cells. Moreover, RRVs appear to carry a heat-labile component that actively suppresses activation of cellular innate immune responses in pDCs. Inhibition of IFN induction by RRVs and the reduced response to IFN should facilitate tumor-specific infection
in vivo
.

IMPORTANCE
RRVs have a convincing preference for replicating in tumor cells in animal models, and we observed similar preferences in the initial treatment of human glioblastoma patients. This study investigates the basis for the interaction between RRV and human host cells (tumor versus nontumor)
in vitro
. We found that RRVs do not trigger an IFN-α/β response in tumor cells, but the cells are capable of responding to type I IFNs and of producing them when stimulated with known agonists. Surprisingly, the data show that RRVs can actively inhibit induction of cellular innate immunity and that this inhibitory activity is heat labile and trypsin sensitive and not attributable to the envelope protein. These data partially explain the observed
in vivo
tumor specificity.
Bilateral anterior lenticonus in a case of Alport syndrome: a clinical and histopathological correlation after successful clear lens extraction
Authors: Sonarkhan Shilpa , Ramappa Muralidhar , Chaurasia Sunita , Mulay Kaustubh
Published: June 2014

A 29-year-old woman presented with symptoms of gradual, progressive, painless diminution of vision in both eyes for past 8–10 years. On examination, uncorrected visual acuity was 20/60 in the right eye and 20/126 in the left eye. Anterior segment examination was essentially normal except for a conical protrusion in the pupillary axis of the anterior lens capsule with a clear underlying lens. During surgery, central protruded fragile lens capsule was meticulously handled to create well-centered continuous curvilinear capsulorhexis by using paediatric rhexis forceps. Histopathology of excised capsule showed markedly thinned lens capsule with normal epithelium. Electron microscopy of the anterior lens capsule showed multiple linear and irregular zones of dehiscence. Few of these had fibrillar, irregular electron-dense material and vacuoles; and adjacent cells were irregular suggestive of Alport syndrome. Postoperative course was uneventful and visual acuity improved to 20/20 unaided. Clear lens extraction is an effective means of rehabilitating visually symptomatic anterior lenticonus of Alport syndrome.
Bilateral ectopic pregnancy following ICSI
Authors: Polat Mehtap , Boynukalın Fazilet Kübra , Yaralı İrem , Yaralı Hakan
Published: June 2014

Bilateral tubal ectopic pregnancy is a rare clinical condition with an estimated prevalence of 1/200 000 spontaneous pregnancies. There is paucity of data on the prevalence of this rare condition following intracytoplasmic sperm injection and embryo transfer (ICSI-ET) cycles. We report two patients with bilateral tubal ectopic pregnancy following ICSI-ET. Both patients had normal, reassuring β-human chorionic gonadotropin dynamics during follow-up; the diagnosis was performed when no gestational sac was noted at the first planned antenatal visit. Of the two patients, one was treated medically and the other surgically with laparoscopic salpingotomy and salpingectomy for the right and left sides, respectively. Both patients thereafter conceived and delivered healthy infants following subsequent ICSI-ET attempts.
The challenges of antiphospholipid syndrome: experience from diagnosis to self-care
Authors: Larmour Kathryn , Lewis Gareth , Benson Gary , Hanko Jennifer
Published: June 2014

A young woman presented to our unit with pancreatitis and acute kidney injury (AKI) 4 weeks after initiation of an oral contraceptive. She subsequently developed seizures due to posterior reversible encephalopathy syndrome and required ongoing haemodialysis for oliguric AKI. Routine antiphospholipid antibody screen was normal, but arterial and venous thromboses were identified on renal biopsy. Further coagulation studies identified an antiphospholipid-dependent inhibitor confirming the suspected diagnosis of antiphospholipid syndrome. She remained seizure free with control of hypertension and was established on anticoagulation. She remained haemodialysis dependent performing this independently at a new self-care unit. She provides us with valuable insights into her experience encouraging us to reconsider our current methods of education and communication in our younger population of patients living with chronic disease.
The mid-Cambrian (Series 3, Guzhangian; Marjuman) trilobite Deiracephalus Resser, 1935, from western Newfoundland
Authors: Westrop Stephen R. , Dengler Alyce A.
Published: June 2014

Species of Deiracephalus Resser, 1935, are rare elements in most Guzhangian (upper Marjuman) trilobite faunas of Laurentian North America, and are characterized by striking cephalic spinosity that includes very long genal and occipital or preoccipital glabellar spines. Almost all previous reports of the genus have assigned sclerites to two species, Deiracephalus aster (Walcott, 1916) and Deiracephalus unicornis Palmer, 1962. However, new material from the Shallow Bay Formation of western Newfoundland and restudy of type material from the southern Appalachians show that as many as eight species are present, although not all of them are sufficiently well known to be formally named. Deiracephalus aster and D. unicornis are restricted to their respective holotypes from the Conasauga Formation of Alabama; new species from the Shallow Bay Formation are Deiracephalus narwhali, Deiracephalus rhinocerotis, Deiracephalus dynastoides, and Deiracephalus phanaeus. Deiracephalus unicornis, D. narwhali, D. rhinocerotis, and a poorly known species from Nevada all possess preoccipital glabellar spines, and form an informal “unicornis group” of species. This group occurs high in the traditional Crepicephalus Zone (latest Guzhangian) and offers a potentially important means of biostratigraphic correlation. Species with occipital spines are older than the unicornis group and extend down into the Cedaria Zone as used traditionally in North America.
Evaluating the effects of liming and wood-ash treatment on forest ecosystems through systematic meta-analysis
Authors: Reid Carolyn , Watmough Shaun A.
Published: June 2014

Liming and wood-ash addition have long been used to attenuate the effects of acidic deposition on forest soils with the goal of promoting tree growth. We performed quantitative meta-analyses of treatment studies from managed forest ecosystems to assess general tendencies of effects of treatment on seven selected measures of performance thought to reasonably reflect the effects of Ca-addition treatment. We retrieved over 350 independent trials from 110 peer-reviewed liming and wood-ash addition studies that were integrated to determine soil pH, base saturation (BS), tree foliar Ca concentration, tree growth, ectomychorrhizae root colonization, soil C-to-N ratio, and microbial indices. The results were quantified through three separate meta-analysis effect size metrics: unweighted relative values and two weighted metrics, Hedges’ d and ln R. A surprising number of treatment trials (22%–85%) reported no significant effect, and soil pH and foliar Ca appeared more responsive to liming than to wood-ash addition, whereas BS and tree growth appeared more responsive to wood-ash addition. For six of the seven parameters, estimated mean effect sizes were similar in magnitude and positive in direction for all three meta-analysis metrics. Regression tree optimal models explained 38% of the variation in pH, 47% of the variation in BS, 51% of the variation in foliar Ca concentration, and 26% of the variation in tree growth. The largest predictors of effect size, within our selected group, were as follows: soil type for pH; soil type, trial duration in years, and species (hardwood or softwood) for BS; treatment dose and type for foliar Ca concentration; and trial duration, initial soil pH, and tree species for tree growth. This analysis shows that Ca additions are not universally beneficial and provides insight into when Ca additions to forest soils are likely to be most effective.
EFFICIENT SCHEME FOR CONGESTION CONTROL IN NETWORK-ON-CHIP WITH QoS CONSIDERATION
Authors: SOUDANI ADEL , ALDAMMAS AHMED , AL-DHELAAN ABDULLAH
Published: June 2014

Embedded distributed multimedia applications based on the use of on-chip networks for communication and messages exchange requires specific and enhanced quality of service (QoS) management. To reach the desired performances at the application level, the network-on-chip (NoC) router should implement per flit handling strategy with wide granularity. This purpose requires an enhanced internal architecture that ensures from one hand a specific management according to a service classification and from the other hand, it enhances the routing process.

In this context, this paper proposes a new mechanism for QoS management in NoC. This mechanism is based on the use of central memory where flits are in-queued according to their class of service. This scheme enables an optimal flit scheduling phase and provides more capabilities to drop low important flits when the router shows congestion state symptoms. The paper presents, also, a protocol structure that fills with this architecture and introduces a signaling mechanism to make efficient the QoS management through the proposed architecture. The circuit performances and its adaptability to achieve QoS with low power processing and high bandwidth in on chip multiprocessor systems will be studied in this paper.
MODELING OF SUPERIOR MESENTERIC ARTERY ANEURYSM USING FLUID–STRUCTURE INTERACTION
Authors: EBRAHIMI BAHARAK , HASSANI KAMRAN
Published: June 2014

The aim of this study was to model the blood flow and predict related hemodynamics characteristics in healthy superior mesenteric artery (SMA) and saccular aneurysm cases. A fluid–structure interaction (FSI) method was performed, using an arbitrary Langrangian–Eulerian mesh. The computational mesh was generated using anatomical data from available human computed tomography (CT)-images. Combining constitution and momentum equations, projection method, the discretized resultant equation were numerically solved for velocity, pressure, shear stress and vortices for healthy/aneurysmal artery. The results including velocity contours, pressure contours, shear rate values, and vortices were obtained and analyzed for three main steps including peak systole, diastole, and end of cardiac cycle. Profiles show the varying velocity and pressure for a pulsatile flow input before and after aneurysms. They also show the formation of single or multiple vortices at aneurysmal area and decrease of wall shear stress with aneurysm enlargement. Furthermore, shear rate values at the neck of aneurysms exceed throughout the entire cardiac cycle. The outcome of the computational analysis is then compared to information available on pressure, vortices and wall shear stress from some clinical findings.
Prediction of rice productivity in Kandhamal district of Orissa using simulation modeling
Authors: Panda Banshidhar , Acharya Milu , Panigrahi Dwitikrishna
Published: June 2014

Simulation technique has been employed to predict rice yield of Kandhamal plateaus in Orissa (India) using the data of previous years. Preliminary simulation model has been developed. The test for uniformity and independence has been conducted using Kolmogrov–smironov test and auto-correlation test, respectively. The result obtained has been subjected to testing of hypothesis by using two sided test. Data for five years (1995 to 2000) are used for model validation and the sample size is increased to 12 years i.e., from 1995 to 2007 for prediction up to 2012. Sensitivity analysis is conducted by changing the parameters within feasible limits to find out the effect on the model.
Identification and lateral membrane localization of cyclin M3, likely to be involved in renal Mg2+handling in seawater fish
Authors: Islam Zinia , Hayashi Naoko , Inoue Hana , Umezawa Takahiro , Kimura Yuuri , Doi Hiroyuki , Romero Michael F. , Hirose Shigehisa , Kato Akira
Published: June 2014

The kidney of marine teleosts is the major site of Mg2+excretion and produces urine with a high Mg2+concentration. However, the transporters involved in Mg2+excretion are poorly understood. The cyclin M (Cnnm; also known as ancient conserved domain protein) family comprises membrane proteins homologous to the bacterial Mg2+and Co2+efflux protein, CorC. To understand the molecular mechanism of Mg2+homeostasis in marine teleosts, we analyzed the expression of the C nnm family genes in the seawater (SW) pufferfish, torafugu ( Takifugu rubripes), and the closely related euryhaline species, mefugu ( Takifugu obscurus). Database mining and phylogenetic analysis indicated that the Takifugu genome contains six members of the Cnnm family: two orthologs of Cnnm1, one of Cnnm2, one of Cnnm3, and two of Cnnm4. RT-PCR analyses indicated that Cnnm2, Cnnm3, and Cnnm4a are expressed in the kidney, whereas other members are mainly expressed in the brain. Renal expression of Cnnm3 was upregulated in SW mefugu, whereas renal expression of Cnnm2 was upregulated in freshwater (FW) mefugu. No significant difference was observed in renal expression of Cnnm4a between SW and FW mefugu. In situ hybridization and immunohistochemical analyses of the SW mefugu kidney revealed that Cnnm3 is expressed in the proximal tubule, and its product localizes to the lateral membrane. When Cnnm3 was expressed in Xenopus laevis oocytes, whole cellular Mg2+content and free intracellular Mg2+activity significantly decreased. These results suggest that Cnnm3 is involved in body fluid Mg2+homeostasis in marine teleosts.
General lysosomal hydrolysis can process prorenin accurately
Authors: Xa Lucie K. , Lacombe Marie-Josée , Mercure Chantal , Lazure Claude , Reudelhuber Timothy L.
Published: June 2014

Renin, an aspartyl protease that catalyzes the rate-limiting step of the renin-angiotensin system, is first synthesized as an inactive precursor, prorenin. Prorenin is activated by the proteolytic removal of an amino terminal prosegment in the dense granules of the juxtaglomerular (JG) cells of the kidney by one or more proteases whose identity is uncertain but commonly referred to as the prorenin-processing enzyme (PPE). Because several extrarenal tissues secrete only prorenin, we tested the hypothesis that the unique ability of JG cells to produce active renin might be explained by the existence of a PPE whose expression is restricted to JG cells. We found that inducing renin production by the mouse kidney by up to 20-fold was not associated with the concomitant induction of candidate PPEs. Because the renin-containing granules of JG cells also contain several lysosomal hydrolases, we engineered mouse Ren1 prorenin to be targeted to the classical vesicular lysosomes of cultured HEK-293 cells, where it was accurately processed and stored. Furthermore, we found that HEK cell lysosomes hydrolyzed any artificial extensions placed on the protein and that active renin was extraordinarily resistant to proteolytic degradation. Altogether, our results demonstrate that accurate processing of prorenin is not restricted to JG cells but can occur in classical vesicular lysosomes of heterologous cells. The implication is that renin production may not require a specific PPE but rather can be achieved by general hydrolysis in the lysosome-like granules of JG cells.