bluuebunny/crossref_metadata_2025 huggingface dataset:
Electroretinographic changes after intravenous lipid emulsion therapy in a dog and a foal with ivermectin toxicosis
Authors: Pollio Danielle , Michau Tammy M. , Weaver Ellen , Kuebelbeck K. Leann
Published: July 2016

This case report describes ivermectin‐induced blindness in a dog and a foal with normal ophthalmic fundic examinations and attenuated electroretinography (ERG). Subsequent recovery inERGwas noted following intravenous lipid emulsion (ILE) therapy. A dog and a foal were evaluated for ivermectin‐induced blindness. Clinical signs included dull mentation, absent pupillary light reflexes (PLRs), and absent menace on presentation. The animals had normal fundoscopic examinations; however, in both casesERGwas consistent with neurosensory retinal dysfunction. FollowingILEtherapy for ivermectin toxicosis, return of menace,PLRs, and normal mentation were noted, as was improvement inERGand serum ivermectin levels. These are the first documented cases of ivermectin‐induced blindness in a dog and a foal with normal fundic examinations and attenuatedERG.ERGimproved in both animals afterILEtherapy.ERGmay assist in the diagnosis of ivermectin toxicosis in dogs and horses.ILEtherapy may hasten recovery in treatment of ivermectin‐induced blindness.
Islet cell transplantation from Göttingen minipigs to cynomolgus monkeys: analysis of virus safety
Authors: Morozov Vladimir A. , Ludwig Stefan , Ludwig Barbara , Rotem Avi , Barkai Uriel , Bornstein Stefan R. , Denner Joachim
Published: July 2016

Background

Xenotransplantation using pig cells, tissues or organs may be associated with the transmission of porcine zoonotic micro‐organisms. Hepatitis E virus (HEV), porcine cytomegalovirus (PCMV) and porcine endogenous retroviruses (PERVs) are potentially zoonotic micro‐organisms which do not show clinical symptoms in pigs and which are due to the low expression level difficult to detect. Göttingen Minipigs (GöMP) are often used for biomedical investigations and they are well characterized concerning the presence of numerous bacteria, fungi, viruses and parasites and therefore may be used for islet cell transplantation.

Methods

Islet cells derived from three GöMP were transplanted into four healthy, non‐diabetic cynomolgus monkeys using a macroencapsulation device. PCR, nested PCR, real‐time PCR, real‐time RT‐PCR and Western blot analyses were used to estimate the presence of PERV, PCMV and HEV in the donors and recipients.

Results

Using sensitive detection methods, no HEV was found in the donor pigs and in the pig islet cell preparations. Antibodies against PERV, PCMV and HEV were not found in all cynomolgus monkeys with exception of one monkey showing an immune response against HEV. Using real‐time PCR, no PCMV and HEV were found in the sera of all monkeys.

Conclusion

Although the donor islet cells and the recipients were negative for HEV using PCR and Western blot analysis, in one recipient, antibodies against HEV were found, indicating infection in a single case. All recipients were negative for antibodies against PERV, and all were negative for PCMV, indicating absence of infection. As HEV was not detected in the donor pig before transplantation, a more complex and regular screening of the animals using highly sensitive methods is required to avoid virus transmission.
Progressive impairment of cerebellar mGluR signalling and its therapeutic potential for cerebellar ataxia in spinocerebellar ataxia type 1 model mice
Authors: Shuvaev Anton N. , Hosoi Nobutake , Sato Yamato , Yanagihara Dai , Hirai Hirokazu
Published: July 2016

Key points

Spinocerebellar ataxia type 1 (SCA1) is a progressive neurodegenerative disease caused by a gene defect, leading to movement disorder such as cerebellar ataxia.

It remains largely unknown which functional defect contributes to the cerebellar ataxic phenotype in SCA1.

In this study, we report progressive dysfunction of metabotropic glutamate receptor (mGluR) signalling, which leads to smaller slow synaptic responses, reduced dendritic Ca2+ signals and impaired synaptic plasticity at cerebellar synapses, in the early disease stage of SCA1 model mice.

We also show that enhancement of mGluR signalling by a clinically available drug, baclofen, leads to improvement of motor performance in SCA1 mice.

SCA1 is an incurable disease with no effective treatment, and our results may provide mechanistic grounds for targeting mGluRs and a novel drug therapy with baclofen to treat SCA1 patients in the future.

Spinocerebellar ataxia type 1 (SCA1) is a progressive neurodegenerative disease that presents with cerebellar ataxia and motor learning defects. Previous studies have indicated that the pathology of SCA1, as well as other ataxic diseases, is related to signalling pathways mediated by the metabotropic glutamate receptor type 1 (mGluR1), which is indispensable for proper motor coordination and learning. However, the functional contribution of mGluR signalling to SCA1 pathology is unclear. In the present study, we show that SCA1 model mice develop a functional impairment of mGluR signalling which mediates slow synaptic responses, dendritic Ca2+ signals, and short‐ and long‐term synaptic plasticity at parallel fibre (PF)–Purkinje cell (PC) synapses in a progressive manner from the early disease stage (5 postnatal weeks) prior to PC death. Notably, impairment of mGluR‐mediated dendritic Ca2+ signals linearly correlated with a reduction of PC capacitance (cell surface area) in disease progression. Enhancement of mGluR signalling by baclofen, a clinically available GABAB receptor agonist, led to an improvement of motor performance in SCA1 mice and the improvement lasted ∼1 week after a single application of baclofen. Moreover, the restoration of motor performance in baclofen‐treated SCA1 mice matched the functional recovery of mGluR‐mediated slow synaptic currents and mGluR‐dependent short‐ and long‐term synaptic plasticity. These results suggest that impairment of synaptic mGluR cascades is one of the important contributing factors to cerebellar ataxia in early and middle stages of SCA1 pathology, and that modulation of mGluR signalling by baclofen or other clinical interventions may be therapeutic targets to treat SCA1.
A Mortar-Based Mesh Interface for Hybrid Finite-Element/Lumped-Parameter Gear Dynamic Models—Applications to Thin-Rimmed Geared Systems
Authors: Guilbert B. , Velex P. , Dureisseix D. , Cutuli P.
Published: July 2016

An original hybrid gear model is introduced, which combines lumped parameter and finite elements along with a specific interface aimed at coupling mismatched discrete models. A mortar-based interface is presented, which eliminates the numerical errors induced by direct collocations between the tooth contact and gear body models. It is shown that the proposed interface can capture the instant contact conditions in the profile and lead directions for both spur and helical gears. A number of quasi-static and dynamic simulation results are presented, which illustrate the potential and practical interest of the methodology. It is observed that thin rims are more influential in the case of helical gears and that the overall dynamic tooth loads seem largely uncoupled from the local contact conditions on the teeth.
The Relationship Between Early-Stage Knee Osteoarthritis and Lower-Extremity Alignment, Joint Laxity, and Subjective Scores of Pain, Stiffness, and Function
Authors: Hicks-Little Charlie A. , Peindl Richard D. , Hubbard-Turner Tricia J. , Cordova Mitchell L.
Published: July 2016

Context:

Knee osteoarthritis (OA) is a debilitating disease that affects an estimated 27 million Americans. Changes in lowerextremity alignment and joint laxity have been found to redistribute the medial and/or lateral loads at the joint. However, the effect that changes in anteroposterior knee-joint laxity have on lower-extremity alignment and function in individuals with knee OA remains unclear.

Objective:

To examine anteroposterior knee-joint laxity, lower-extremity alignment, and subjective pain, stiffness, and function scores in individuals with early-stage knee OA and matched controls and to determine if a relationship exists among these measures.

Design:

Case control.

Setting:

Sports-medicine research laboratory.

Participants:

18 participants with knee OA and 18 healthy matched controls.

Intervention:

Participants completed the Western Ontario McMaster (WOMAC) osteoarthritis questionnaire and were tested for total anteroposterior knee-joint laxity (A-P) and knee-joint alignment (ALIGN).

Main Outcome Measures:

WOMAC scores, A-P (mm), and ALIGN (°).

Results:

A significant multivariate main effect for group (Wilks’ Λ = 0.30, F7,26 = 8.58, P < .0001) was found. Knee-OA participants differed in WOMAC scores (P < .0001) but did not differ from healthy controls on ALIGN (P = .49) or total A-P (P = .66). No significant relationships were identified among main outcome measures.

Conclusion:

These data demonstrate that participants with early-stage knee OA had worse pain, stiffness, and functional outcome scores than the matched controls; however, ALIGN and A-P were no different. There was no association identified among participants’ subjective scores, ALIGN, or A-P measures in this study.
Associations Between Functional and Isolated Performance Measures in College Women’s Soccer Players
Authors: McCann Ryan S. , Kosik Kyle B. , Terada Masafumi , Beard Megan Q. , Buskirk Gretchen E. , Gribble Phillip A.
Published: July 2016

Context:

The Star Excursion Balance Test (SEBT) and Functional Movement Screen (FMS) are functional performance measures capable of predicting lower-extremity injury risk. While suboptimal SEBT and FMS performances are influenced by multiple factors, the contribution of hip strength and flexibility to these tests is mostly unknown. Examination of hip strength and flexibility influences on the SEBT and FMS may direct clinicians to better methods of correcting functional deficits.

Objective:

Determine the relationships of isometric hip strength and hip passive range of motion (PROM) with functional performance measures.

Design:

Cross-sectional.

Setting:

Athletic training facility.

Participants:

43 NCAA Division I women’s soccer players (19.65 ± 1.12 y; 166.93 ± 3.84 cm; 60.99 ± 4.31 kg) volunteered.

Data Collection and Analysis:

All participants were tested bilaterally in the SEBT; the deep squat, in-line lunge, hurdle step, and straight leg raise, comprising a lower-extremity FMS (FMS-LE); hip internal and external rotation PROM; and isometric hip extension strength (HEXT). The mean of the 3 averaged, normalized SEBT scores was used as a composite score. Pearson product moment correlations assessed relationships of SEBT and FMS-LE scores with PROM and HEXT. Significance was set a priori at P < .05.

Results:

Pearson correlations revealed anterior (ANT) SEBT scores had a low negative association with HEXT (r = –0.33,P = .004) and a low positive association with hip internal rotation PROM (PROM-IR) (r = .43,P = .003). All other correlations were negligible.

Conclusions:

Flexibility training aimed at PROM-IR may contribute to improved ANT scores. Targeting HEXT and hip external rotation PROM are likely not preferred means of correcting deficits in SEBT and FMS-LE performance.
The Functional Movement Screen in the Prediction of Injury in Adolescent Cricket Pace Bowlers: An Observational Study
Authors: Martin Candice , Olivier Benita , Benjamin Natalie
Published: July 2016

Context:

The Functional Movement Screen (FMS) has been found to be a valid preparticipation screening tool in the prediction of injury among various athletes in different sports. The validity thereof in the prediction of injury among adolescent cricketers is yet to be established.

Objective:

To determine if a preseason FMS total score is a valid predictor of in-season injury among adolescent pace bowlers.

Design:

Prospective observational quantitative study.

Setting:

Bowlers performed the FMS before the start of the season. Injury incidence was monitored monthly throughout the season. The student t test and Fisher’s exact test were used to compare the FMS scores of the injured and noninjured bowlers as well as the injured and noninjured bowlers who scored ≤ 14.

Participants:

27 injury-free, male, adolescent pace bowlers.

Main Outcome Measures:

The FMS (scoring criteria and score sheet) and standardized self-administered injury questionnaire.

Results:

There was no difference between the noninjured group (16.55 ± 2.57) and the injured (16.1 ± 2.07) group in terms of FMS scores. There was no significant difference between injured and noninjured bowlers who scored ≤ 14. A total FMS score of 14 does not provide the sensitivity needed to assess injury risk among adolescent pace bowlers and no other accurate cut-off score could be calculated.

Conclusion:

Preseason observed total FMS score is a poor predictor of in-season injury among adolescent pace bowlers. Further research should be conducted to determine if a specific FMS test will be a more valid predictor of injury.
Protein engineering by highly parallel screening of computationally designed variants
Authors: Sun Mark G. F. , Seo Moon-Hyeong , Nim Satra , Corbi-Verge Carles , Kim Philip M.
Published: July 2016

Combining the power of combinatorial screening and computational design for optimal protein engineering.
To Conquer the Host, Influenza Virus Is Packing It In: Interferon-Antagonistic Strategies beyond NS1
Authors: Weber-Gerlach Michaela , Weber Friedemann
Published: July 2016

The nonstructural protein NS1 is well established as a virulence factor of influenza A virus counteracting induction of the antiviral type I interferon system. Recent studies now show that viral structural proteins, their derivatives, and even the genome itself also contribute to keeping the host defense under control. Here, we summarize the current knowledge on these NS1-independent interferon escape strategies.
Reversion of Cold-Adapted Live Attenuated Influenza Vaccine into a Pathogenic Virus
Authors: Zhou Bin , Meliopoulos Victoria A. , Wang Wei , Lin Xudong , Stucker Karla M. , Halpin Rebecca A. , Stockwell Timothy B. , Schultz-Cherry Stacey , Wentworth David E.
Published: July 2016

The only licensed live attenuated influenza A virus vaccines (LAIVs) in the United States (FluMist) are created using internal protein-coding gene segments from the cold-adapted temperature-sensitive master donor virus A/Ann Arbor/6/1960 and HA/NA gene segments from circulating viruses. During serial passage of A/Ann Arbor/6/1960 at low temperatures to select the desired attenuating phenotypes, multiple cold-adaptive mutations and temperature-sensitive mutations arose. A substantial amount of scientific and clinical evidence has proven that FluMist is safe and effective. Nevertheless, no study has been conducted specifically to determine if the attenuating temperature-sensitive phenotype can revert and, if so, the types of substitutions that will emerge (i.e., compensatory substitutions versus reversion of existing attenuating mutations). Serial passage of the monovalent FluMist 2009 H1N1 pandemic vaccine at increasing temperatures
in vitro
generated a variant that replicated efficiently at higher temperatures. Sequencing of the variant identified seven nonsynonymous mutations, PB1-E51K, PB1-I171V, PA-N350K, PA-L366I, NP-N125Y, NP-V186I, and NS2-G63E. None occurred at positions previously reported to affect the temperature sensitivity of influenza A viruses. Synthetic genomics technology was used to synthesize the whole genome of the virus, and the roles of individual mutations were characterized by assessing their effects on RNA polymerase activity and virus replication kinetics at various temperatures. The revertant also regained virulence and caused significant disease in mice, with severity comparable to that caused by a wild-type 2009 H1N1 pandemic virus.

IMPORTANCE
The live attenuated influenza vaccine FluMist has been proven safe and effective and is widely used in the United States. The phenotype and genotype of the vaccine virus are believed to be very stable, and mutants that cause disease in animals or humans have never been reported. By propagating the virus under well-controlled laboratory conditions, we found that the FluMist vaccine backbone could regain virulence to cause severe disease in mice. The identification of the responsible substitutions and elucidation of the underlying mechanisms provide unique insights into the attenuation of influenza virus, which is important to basic research on vaccines, attenuation reversion, and replication. In addition, this study suggests that the safety of LAIVs should be closely monitored after mass vaccination and that novel strategies to continue to improve LAIV vaccine safety should be investigated.
Cryo-electron Microscopy Reconstruction and Stability Studies of the Wild Type and the R432A Variant of Adeno-associated Virus Type 2 Reveal that Capsid Structural Stability Is a Major Factor in Genome Packaging
Authors: Drouin Lauren M. , Lins Bridget , Janssen Maria , Bennett Antonette , Chipman Paul , McKenna Robert , Chen Weijun , Muzyczka Nicholas , Cardone Giovanni , Baker Timothy S. , Agbandje-McKenna Mavis
Published: July 2016

The adeno-associated viruses (AAV) are promising therapeutic gene delivery vectors and better understanding of their capsid assembly and genome packaging mechanism is needed for improved vector production. Empty AAV capsids assemble in the nucleus prior to genome packaging by virally encoded Rep proteins. To elucidate the capsid determinants of this process, structural differences between wild-type (wt) AAV2 and a packaging deficient variant, AAV2-R432A, were examined using cryo-electron microscopy and three-dimensional image reconstruction both at an ∼5.0-Å resolution (medium) and also at 3.8- and 3.7-Å resolutions (high), respectively. The high resolution structures showed that removal of the arginine side chain in AAV2-R432A eliminated hydrogen bonding interactions, resulting in altered intramolecular and intermolecular interactions propagated from under the 3-fold axis toward the 5-fold channel. Consistent with these observations, differential scanning calorimetry showed an ∼10°C decrease in thermal stability for AAV2-R432A compared to wt-AAV2. In addition, the medium resolution structures revealed differences in the juxtaposition of the less ordered, N-terminal region of their capsid proteins, VP1/2/3. A structural rearrangement in AAV2-R432A repositioned the βA strand region under the icosahedral 2-fold axis rather than antiparallel to the βB strand, eliminating many intramolecular interactions. Thus, a single amino acid substitution can significantly alter the AAV capsid integrity to the extent of reducing its stability and possibly rendering it unable to tolerate the stress of genome packaging. Furthermore, the data show that the 2-, 3-, and 5-fold regions of the capsid contributed to producing the packaging defect and highlight a tight connection between the entire capsid in maintaining packaging efficiency.

IMPORTANCE
The mechanism of AAV genome packaging is still poorly understood, particularly with respect to the capsid determinants of the required capsid-Rep interaction. Understanding this mechanism may aid in the improvement of AAV packaging efficiency, which is currently ∼1:10 (10%) genome packaged to empty capsid in vector preparations. This report identifies regions of the AAV capsid that play roles in genome packaging and that may be important for Rep recognition. It also demonstrates the need to maintain capsid stability for the success of this process. This information is important for efforts to improve AAV genome packaging and will also inform the engineering of AAV capsid variants for improved tropism, specific tissue targeting, and host antibody escape by defining amino acids that cannot be altered without detriment to infectious vector production.
Human Cytomegalovirus Can Procure Deoxyribonucleotides for Viral DNA Replication in the Absence of Retinoblastoma Protein Phosphorylation
Authors: Kuny Chad V. , Kalejta Robert F.
Published: July 2016

Viral DNA replication requires deoxyribonucleotide triphosphates (dNTPs). These molecules, which are found at low levels in noncycling cells, are generated either by salvage pathways or through
de novo
synthesis. Nucleotide synthesis utilizes the activity of a series of nucleotide-biosynthetic enzymes (NBEs) whose expression is repressed in noncycling cells by complexes between the E2F transcription factors and the retinoblastoma (Rb) tumor suppressor. Rb-E2F complexes are dissociated and NBE expression is activated during cell cycle transit by cyclin-dependent kinase (Cdk)-mediated Rb phosphorylation. The DNA virus human cytomegalovirus (HCMV) encodes a viral Cdk (v-Cdk) (the UL97 protein) that phosphorylates Rb, induces the expression of cellular NBEs, and is required for efficient viral DNA synthesis. A long-held hypothesis proposed that viral proteins with Rb-inactivating activities functionally similar to those of UL97 facilitated viral DNA replication in part by inducing the
de novo
production of dNTPs. However, we found that dNTPs were limiting even in cells infected with wild-type HCMV in which UL97 is expressed and Rb is phosphorylated. Furthermore, we revealed that both
de novo
and salvage pathway enzymes contribute to viral DNA replication during HCMV infection and that Rb phosphorylation by cellular Cdks does not correct the viral DNA replication defect observed in cells infected with a UL97-deficient virus. We conclude that HCMV can obtain dNTPs in the absence of Rb phosphorylation and that UL97 can contribute to the efficiency of DNA replication in an Rb phosphorylation-independent manner.

IMPORTANCE
Transforming viral oncoproteins, such as adenovirus E1A and papillomavirus E7, inactivate Rb. The standard hypothesis for how Rb inactivation facilitates infection with these viruses is that it is through an increase in the enzymes required for DNA synthesis, which include nucleotide-biosynthetic enzymes. However, HCMV UL97, which functionally mimics these viral oncoproteins through phosphorylation of Rb, fails to induce the production of nonlimiting amounts of dNTPs. This finding challenges the paradigm of the role of Rb inactivation during DNA virus infection and uncovers the existence of an alternative mechanism by which UL97 contributes to HCMV DNA synthesis. The ineffectiveness of the UL97 inhibitor maribavir in clinical trials might be better explained with a fuller understanding of the role of UL97 during infection. Furthermore, as the nucleoside analog ganciclovir is the current drug of choice for treating HCMV, knowing the provenance of the dNTPs incorporated into viral DNA may help inform antiviral therapeutic regimens.
HIV-1 Vpr Inhibits Kaposi's Sarcoma-Associated Herpesvirus Lytic Replication by Inducing MicroRNA miR-942-5p and Activating NF-κB Signaling
Authors: Yan Qin , Shen Chenyou , Qin Jie , Li Wan , Hu Minmin , Lu Hongmei , Qin Di , Zhu Jianzhong , Gao Shou-Jiang , Lu Chun
Published: July 2016

Kaposi's sarcoma-associated herpesvirus (KSHV) infection is required for the development of several AIDS-related malignancies, including Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). The high incidence of AIDS-KS has been ascribed to the interaction of KSHV and HIV-1. We have previously shown that HIV-1-secreted proteins Tat and Nef regulate the KSHV life cycle and synergize with KSHV oncogenes to promote angiogenesis and tumorigenesis. Here, we examined the regulation of KSHV latency by HIV-1 viral protein R (Vpr). We found that soluble Vpr inhibits the expression of KSHV lytic transcripts and proteins, as well as viral particle production by activating NF-κB signaling following internalization into PEL cells. By analyzing the expression profiles of microRNAs combined with target search by bioinformatics and luciferase reporter analyses, we identified a Vpr-upregulated cellular microRNA (miRNA), miR-942-5p, that directly targeted IκBα. Suppression of miR-942-5p relieved the expression of IκBα and reduced Vpr inhibition of KSHV lytic replication, while overexpression of miR-942-5p enhanced Vpr inhibition of KSHV lytic replication. Our findings collectively illustrate that, by activating NF-κB signaling through upregulating a cellular miRNA to target IκBα, internalized HIV-1 Vpr inhibits KSHV lytic replication. These results have demonstrated an essential role of Vpr in the life cycle of KSHV.

IMPORTANCE
Coinfection by HIV-1 promotes the aggressive growth of Kaposi's sarcoma-associated herpesvirus (KSHV)-related malignancies, including Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). In this study, we have shown that soluble HIV-1 Vpr inhibits KSHV lytic replication by activating NF-κB signaling following internalization into PEL cells. Mechanistic studies revealed that a cellular microRNA upregulated by Vpr, miR-942-5p, directly targeted IκBα. Suppression of miR-942-5p relieved IκBα expression and reduced Vpr inhibition of KSHV replication, while overexpression of miR-942-5p enhanced Vpr inhibition of KSHV replication. These results indicate that by activating NF-κB signaling through upregulating a cellular miRNA to target IκBα, internalized Vpr inhibits KSHV lytic replication. This work illustrates a molecular mechanism by which HIV-1-secreted regulatory protein Vpr regulates KSHV latency and the pathogenesis of AIDS-related malignancies.
Relationship between Vaccine-Induced Antibody Capture of Infectious Virus and Infection Outcomes following Repeated Low-Dose Rectal Challenges with Simian Immunodeficiency Virus SIVmac251
Authors: Gach Johannes S. , Venzon David , Vaccari Monica , Keele Brandon F. , Franchini Genoveffa , Forthal Donald N.
Published: July 2016

Antibodies are known to enhance
in vitro
infection by human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV). We measured the ability of antibodies induced by ALVAC-SIV/gp120 vaccination, given with alum or MF59 adjuvant, to capture infectious SIVmac251 and determined the association between capture and infection outcomes following low-dose, repeated rectal challenge of rhesus macaques. We found that capture correlated with the number of transmitted/founder (T/F) variants that established infection, such that animals whose plasma captured more virus were infected with a higher number of T/F strains. Capture also correlated with results of Env binding assays, indicating that greater immunogenicity resulted in greater capture. Although vaccination elicited negligible neutralizing activity against the challenge strain (50% inhibitory dilutions of >1/80 in all cases), animals with low capture and whose plasma, at a fixed dilution, inhibited a higher fraction of virus were infected at a lower rate than animals with high capture and low neutralization (
P
= 0.039); only animals with the low capture/high neutralization response profile were protected compared with unvaccinated control animals (
P
= 0.026). In a sieve analysis, high capture and low capture were distinguishable on the basis of polymorphisms in the V1 loop of Env at amino acids 144 and 145. Our results indicate that vaccine-induced antibody that binds to and captures infectious virus but does not inhibit its infectivity may enhance the likelihood of infection following rectal challenge with SIVmac251. Higher immunogenicity resulting in better antibody capture but similar anti-infectivity may not improve vaccine efficacy.

IMPORTANCE
Vaccines generally prevent viral infections by eliciting antibodies that inhibit virus infectivity. However, antibodies, including those induced by vaccination, have the potential to enhance, rather than prevent infection. We measured the ability of vaccine-induced antibodies to capture infectious simian immunodeficiency virus (SIV) and explored the relationship between virus capture and infection outcomes. We found that capture correlated with the number of SIV variants that established infection, such that animals whose plasma captured more virus were infected with a higher number of unique strains. In addition, animals whose sera had high capture but weak anti-infectivity activity were infected at a higher rate than were animals with low capture and stronger anti-infectivity activity. These results suggest that vaccines that induce antibodies that bind to and capture infectious virus but do not inhibit virus infectivity will not be effective in preventing infection.
Unmasking Stem-Specific Neutralizing Epitopes by Abolishing N-Linked Glycosylation Sites of Influenza Virus Hemagglutinin Proteins for Vaccine Design
Authors: Liu Wen-Chun , Jan Jia-Tsrong , Huang Yun-Ju , Chen Ting-Hsuan , Wu Suh-Chin
Published: July 2016

Influenza virus hemagglutinin (HA) protein consists of two components, i.e., a globular head region and a stem region that are folded within six disulfide bonds, plus several N-linked glycans that produce a homotrimeric complex structure. While N-linked glycosylation sites on the globular head are variable among different strains and different subtypes, N-linked glycosylation sites in the stem region are mostly well conserved among various influenza virus strains. Targeting highly conserved HA stem regions has been proposed as a useful strategy for designing universal influenza vaccines. Since the HA stem region is constituted by an HA1 N-terminal part and a full HA2 part, we expressed a series of recombinant HA mutant proteins with deleted N-linked glycosylation sites in the HA1 stem and HA2 stem regions of H5N1 and pH1N1 viruses. Unmasking N-glycans in the HA2 stem region (H5 N484A and H1 N503A) was found to elicit more potent neutralizing antibody titers against homologous, heterologous, and heterosubtypic viruses. Unmasking the HA2 stem N-glycans of H5HA but not H1HA resulted in more CR6261-like and FI6v3-like antibodies and also correlated with the increase of cell fusion inhibition activity in antisera. Only H5 N484A HA2 stem mutant protein immunization increased the numbers of antibody-secreting cells, germinal center B cells, and memory B cells targeting the stem helix A epitopes in splenocytes. Unmasking the HA2 stem N-glycans of H5HA mutant proteins showed a significantly improvement in the protection against homologous virus challenges but did so to a less degree for the protection against heterosubtypic pH1N1 virus challenges. These results may provide useful information for designing more effective influenza vaccines.

IMPORTANCE
N-linked glycosylation sites in the stem regions of influenza virus hemagglutinin (HA) proteins are mostly well conserved among various influenza virus strains. Targeting highly conserved HA stem regions has been proposed as a useful strategy for designing universal influenza vaccines. Our studies indicate that unmasking the HA2 stem N-glycans of recombinant HA proteins from H5N1 and pH1N1 viruses induced more potent neutralizing antibody titers against homologous and heterosubtypic viruses. However, only immunization with the H5N1 HA2 stem mutant protein can refocus B antibody responses to the helix A epitope for inducing more CR6261-like/FI6v3-like and fusion inhibition antibodies in antisera, resulting in a significant improvement for the protection against lethal H5N1 virus challenges. These results may provide useful information for designing more effective influenza vaccines.
Varicella-Zoster Virus Activates CREB, and Inhibition of the pCREB-p300/CBP Interaction Inhibits Viral ReplicationIn Vitroand Skin PathogenesisIn Vivo
Authors: François Sylvie , Sen Nandini , Mitton Bryan , Xiao Xiangshu , Sakamoto Kathleen M. , Arvin Ann
Published: July 2016

Varicella-zoster virus (VZV) is an alphaherpesvirus that causes varicella upon primary infection and zoster upon reactivation from latency in sensory ganglion neurons. The replication of herpesviruses requires manipulation of cell signaling pathways. Notably, CREB, a factor involved in the regulation of several cellular processes, is activated upon infection of T cells with VZV. Here, we report that VZV infection also induced CREB phosphorylation in fibroblasts and that XX-650-23, a newly identified inhibitor of the phosphorylated-CREB (pCREB) interaction with p300/CBP, restricted cell-cell spread of VZVin vitro. CREB phosphorylation did not require the viral open reading frame 47 (ORF47) and ORF66 kinases encoded by VZV. Evaluating the biological relevance of these observations during VZV infection of human skin xenografts in the SCID mouse model of VZV pathogenesis showed both that pCREB was upregulated in infected skin and that treatment with XX-650-23 reduced infectious-virus production and limited lesion formation compared to treatment with a vehicle control. Thus, processes of CREB activation and p300/CBP binding are important for VZV skin infection and may be targeted for antiviral drug development.

IMPORTANCEVaricella-zoster virus (VZV) is a common pathogen that causes chicken pox and shingles. As with all herpesviruses, the infection is acquired for life, and the virus can periodically reactivate from latency. Although VZV infection is usually benign with few or no deleterious consequences, infection can be life threatening in immunocompromised patients. Otherwise healthy elderly individuals who develop zoster as a consequence of viral reactivation are at risk for postherpetic neuralgia (PHN), a painful and long-lasting complication. Current vaccines use a live attenuated virus that is usually safe but cannot be given to many immunodeficient patients and retains the capacity to establish latency and reactivate, causing zoster. Antiviral drugs are effective against severe VZV infections but have little impact on PHN. A better understanding of virus-host cell interactions is relevant for developing improved therapies to safely interfere with cellular processes that are crucial for VZV pathogenesis.
Hepatitis B Virus Polymerase Localizes to the Mitochondria, and Its Terminal Protein Domain Contains the Mitochondrial Targeting Signal
Authors: Unchwaniwala Nuruddin , Sherer Nathan M. , Loeb Daniel D.
Published: July 2016

To understand subcellular sites of hepatitis B virus (HBV) replication, we visualized core (Cp), polymerase (Pol), and pregenomic RNA (pgRNA) in infected cells. Interestingly, we found that the majority of Pol localized to the mitochondria in cells undergoing viral replication. The mitochondrial localization of Pol was independent of both the cell type and other viral components, indicating that Pol contains an intrinsic mitochondrial targeting signal (MTS). Neither Cp nor pgRNA localized to the mitochondria during active replication, suggesting a role other than DNA synthesis for Pol at the mitochondria. The Pol of duck hepatitis B virus (DHBV) also localized to the mitochondria. This result indicates that localization of Pol to mitochondria is likely a feature of all hepadnaviruses. To map the MTS within HBV Pol, we generated a series of Pol-green fluorescent protein (Pol-GFP) fusions and found that a stretch spanning amino acids (aa) 141 to 160 of Pol was sufficient to target GFP to the mitochondria. Surprisingly, deleting aa 141 to 160 in full-length Pol did not fully ablate Pol's mitochondrial localization, suggesting that additional sequences are involved in mitochondrial targeting. Only by deleting the N-terminal 160 amino acids in full-length Pol was mitochondrial localization ablated. Crucial residues for pgRNA packaging are contained within aa 141 to 160, indicating a multifunctional role of this region of Pol in the viral life cycle. Our studies show an unexpected Pol trafficking behavior that is uncoupled from its role in viral DNA synthesis.

IMPORTANCE
Chronic infection by HBV is a serious health concern. Existing therapies for chronically infected individuals are not curative, underscoring the need for a better understanding of the viral life cycle to develop better antiviral therapies. To date, the most thoroughly studied function of Pol is to package the pgRNA and reverse transcribe it to double-stranded DNA within capsids. This study provides evidence for mitochondrial localization of Pol and defines the MTS. Recent findings have implicated a non-reverse transcription role for Pol in evading host innate immune responses. Mitochondria play an important role in controlling cellular metabolism, apoptosis, and innate immunity. Pol may alter one or more of these host mitochondrial functions to gain a replicative advantage and persist in chronically infected individuals.
An Ebola Virus-Like Particle-Based Reporter System Enables Evaluation of Antiviral Drugs
In Vivo
under Non-Biosafety Level 4 Conditions
Authors: Li Dapeng , Chen Tan , Hu Yang , Zhou Yu , Liu Qingwei , Zhou Dongming , Jin Xia , Huang Zhong
Published: July 2016

Ebola virus (EBOV) is a highly contagious lethal pathogen. As a biosafety level 4 (BSL-4) agent, however, EBOV is restricted to costly BSL-4 laboratories for experimentation, thus significantly impeding the evaluation of EBOV vaccines and drugs. Here, we report an EBOV-like particle (EBOVLP)-based luciferase reporter system that enables the evaluation of anti-EBOV agents
in vitro
and
in vivo
outside BSL-4 facilities. Cotransfection of HEK293T cells with four plasmids encoding the proteins VP40, NP, and GP of EBOV and firefly luciferase (Fluc) resulted in the production of Fluc-containing filamentous particles that morphologically resemble authentic EBOV. The reporter EBOVLP was capable of delivering Fluc into various cultured cells in a GP-dependent manner and was recognized by a conformation-dependent anti-EBOV monoclonal antibody (MAb). Significantly, inoculation of mice with the reporter EBOVLP led to the delivery of Fluc protein into target cells and rapid generation of intense bioluminescence signals that could be blocked by the administration of EBOV neutralizing MAbs. This BSL-4-free reporter system should facilitate high-throughput screening for anti-EBOV drugs targeting viral entry and efficacy testing of candidate vaccines.

IMPORTANCE
Ebola virus (EBOV) researches have been limited to costly biosafety level 4 (BSL-4) facilities due to the lack of animal models independent of BSL-4 laboratories. In this study, we reveal that a firefly luciferase-bearing EBOV-like particle (EBOVLP) with typical filamentous EBOV morphology is capable of delivering the reporter protein into murine target cells both
in vitro
and
in vivo
. Moreover, we demonstrate that the reporter delivery can be inhibited both
in vitro
and
in vivo
by a known anti-EBOV protective monoclonal antibody, 13C6. Our work provides a BSL-4-free system that can facilitate the
in vivo
evaluation of anti-EBOV antibodies, drugs, and vaccines. The system may also be useful for mechanistic study of the viral entry process.
Investigating the association between prepregnancy body mass index and adverse pregnancy outcomes: a large cohort study of 536 098 Chinese pregnant women in rural China
Authors: Pan Yi , Zhang Shikun , Wang Qiaomei , Shen Haiping , Zhang Yiping , Li Yuanyuan , Yan Donghai , Sun Lizhou
Published: July 2016

Objective

Unhealthy maternal weight before pregnancy increases the risk of various adverse pregnancy outcomes. We conducted a nutrition survey to provide baseline data on the prepregnant nutritional status of mothers in order to better understand the association between prepregnancy maternal body mass index (BMI) and adverse pregnancy outcomes.

Design

A large, prospective, population-based cohort study.

Setting

Data from the National Free Preconception Health Examination Project (NFPHEP) in China during 2010–2012.

Participants

536 098 pregnant women out of 2 120 131 were evaluated.

Primary and secondary outcome measures

The primary adverse pregnancy outcomes included preterm birth (PTB), low birth weight (LBW), spontaneous miscarriage (SM), ectopic pregnancy (EP) and stillbirth (SB). A χ2test was used to compare the prevalence of each BMI category during 2010–2012. Univariable and multiple logistic regression analyses were performed to assess the association between prepregnancy BMI and various adverse pregnancy outcomes.

Results

Between 2010 and 2012, the average BMI decreased from 21.31 to 21.16, while underweight prevalence increased from 10.40% to 14.14%. An age-stratified subgroup analysis indicated that the underweight prevalence increased from 13.52% to 17.02% among women aged 21–24 and from 10.72% to 13.71% among women aged 25–34. Overweight prevalence increased from 9.84% to 10.75% (25–34 years) and from 17.10% to 19.20% (35–49 years). Obesity prevalence increased from 2.17% to 2.42% and from 4% to 4.2% among women aged 25–34 and 35–49 respectively. Prepregnancy underweight was associated with PTB, LBW and SM; overweight women had an increased risk of LBW; obese women had a higher risk of LBW, SM, EP and SB.

Conclusions

While the average prepregnancy BMI decreased, the prevalence of underweight individuals in a very large population significantly increased. The abnormal prepregnancy BMIs were associated with increased risks of adverse pregnancy outcomes. Most notably, underweight prepregnant women appeared to be at a greater risk of developing adverse pregnancy outcomes in China's rural areas.
Glycated haemoglobin (HbA1c), diabetes and trajectories of change in episodic memory performance
Authors: Pappas Colleen , Andel Ross , Infurna Frank J , Seetharaman Shyam
Published: July 2016

Background

As the ageing population grows, it is important to identify strategies to moderate cognitive ageing.

Objective

We examined glycated haemoglobin (HbA1c) and diabetes in relation to level and change in episodic memory in older adults with and without diabetes.

Methods

Data from 4419 older adults with (n=950) and without (n=3469) diabetes participating in a nationally representative longitudinal panel study (the Health and Retirement Study) were examined. Average baseline age was 72.66 years and 58% were women. HbA1c was measured in 2006 and episodic memory was measured using immediate and delayed list recall over 4 biennial waves between 2006 and 2012. Growth curve models were used to assess trajectories of episodic memory change.

Results

In growth curve models adjusted for age, sex, education, race, depressive symptoms and waist circumference, higher HbA1c levels and having diabetes were associated with poorer baseline episodic memory (p=0.036 and <0.001, respectively) and greater episodic memory decline (p=0.006 and 0.004, respectively). The effect of HbA1c on episodic memory decline was smaller than the effect of age. The results were stronger for women than men and were not modified by age or race. When the main analyses were estimated for those with and without diabetes separately, HbA1c was significantly linked to change in episodic memory only among those with diabetes.

Conclusions

Higher HbA1c and diabetes were both associated with declines in episodic memory, with this relationship further exacerbated by having diabetes and elevated HbA1c. HbA1c appeared more important for episodic memory performance among women than men.
Republished: Transvenous ethanol sclerotherapy of feeding arteries for treatment of a dural arteriovenous fistula
Authors: Alexander Matthew David , Halbach Van , Nicholson Andrew , Settecase Fabio , Darflinger Robert J , Amans Matthew R
Published: July 2016

Dural arteriovenous fistulas (DAVFs) can be complex lesions that require a well trained eye for proper characterization and management decisions. With numerous possible arteries to supply and veins to drain them, DAVFs are often complex lesions. To best treat these complex lesions, the neurointerventionalist should be adept at treating them with multiple techniques. This report describes a unique case in which a DAVF was treated with arterial sclerotherapy using ethanol from a transvenous approach.
Mid-term and long-term follow-up of intracranial aneurysms treated by the p64 Flow Modulation Device: a multicenter experience
Authors: Briganti Francesco , Leone Giuseppe , Ugga Lorenzo , Marseglia Mariano , Macera Antonio , Manto Andrea , Delehaye Luigi , Resta Maurizio , Resta Mariachiara , Burdi Nicola , Nuzzi Nunzio Paolo , Divenuto Ignazio , Caranci Ferdinando , Muto Mario , Solari Domenico , Cappabianca Paolo , Maiuri Francesco
Published: July 2016

Background

Experience with the endovascular treatment of cerebral aneurysms using the p64 Flow Modulation Device is still limited. This study discusses the results and complications of this new flow diverter device.

Methods

40 patients (30 women, 10 men) with 50 cerebral aneurysms treated in six Italian neurointerventional centers with the p64 Flow Modulation Device between April 2013 and September 2015 were retrospectively reviewed.

Results

Complete occlusion was obtained in 44/50 aneurysms (88%) and partial occlusion in 3 (6%). In the other three aneurysms (6%), two cases of asymptomatic in-stent thrombosis and one intraprocedural occlusion of the parent vessel occurred. Technical complications were observed in eight procedures (16%). Permanent morbidity due to acute in-stent thrombosis and consequent ischemic stroke occurred in one patient (2.5%). No delayed aneurysm rupture, subarachnoid or intraparenchymal hemorrhage, or ischemic complications occurred and there were no deaths.

Conclusions

Endovascular treatment with the p64 Flow Modulation Device is a safe treatment for unruptured cerebral aneurysms, resulting in a high rate of occlusion. As with other flow diverter devices, we recommend this treatment mainly for large-necked aneurysms of the internal carotid artery siphon. However, endovascular treatment with the p64 device should also be encouraged in difficult cases such as aneurysms of the posterior circulation and beyond the circle of Willis.
Thermal constraints on stream consumer responses to a marine resource subsidy
Authors: Smits Adrianne P. , Schindler Daniel E. , Armstrong Jonathan B. , Brett Michael T. , Carter Jackie L. , Santos Bianca S.
Published: July 2016

Spawning migrations of Pacific salmon (Oncorhynchus spp.) to coastal watersheds provide a rich resource subsidy to freshwater consumers. However, variation in thermal regimes and spawning activity across the landscape constrain the ability of poikilothermic consumers to assimilate eggs and carcasses. We investigated how sockeye salmon (Oncorhynchus nerka) spawning density and stream temperature affect the growth, body condition, and fatty acid composition of juvenile coho salmon (Oncorhynchus kisutch), a known egg predator, in seven tributaries of the Wood River in Southwest Alaska. We compared mean body size of juvenile coho salmon in late summer among 3–7 years per stream and found that the largest mean size occurred in warm streams in which sockeye salmon spawned, although overall subsidy magnitude (spawner density) had no effect on consumer body size. Individuals that consumed more salmon eggs (estimated from δ15N) were larger and had altered fatty acid composition but did not have higher relative body condition. These results indicate that effects of marine subsidies on freshwater consumers depend both on local habitat conditions and on individual variation in energy allocation.
Higher U(1)-gerbe connections in geometric prequantization
Authors: Fiorenza Domenico , Rogers Christopher L. , Schreiber Urs
Published: July 2016

We promote geometric prequantization to higher geometry (higher stacks), where a prequantization is given by a higher principal connection (a higher gerbe with connection). We show fairly generally how there is canonically a tower of higher gauge groupoids and Courant groupoids assigned to a higher prequantization, and establish the corresponding Atiyah sequence as an integrated Kostant–Souriau [Formula: see text]-group extension of higher Hamiltonian symplectomorphisms by higher quantomorphisms. We also exhibit the [Formula: see text]-group cocycle which classifies this extension and discuss how its restrictions along Hamiltonian [Formula: see text]-actions yield higher Heisenberg cocycles. In the special case of higher differential geometry over smooth manifolds, we find the [Formula: see text]-algebra extension of Hamiltonian vector fields — which is the higher Poisson bracket of local observables — and show that it is equivalent to the construction proposed by the second author in [Formula: see text]-plectic geometry. Finally, we indicate a list of examples of applications of higher prequantization in the extended geometric quantization of local quantum field theories and specifically in string geometry.