bluuebunny/crossref_metadata_2025 huggingface dataset:
Attention and Temporal Expectations Modulate Power, Not Phase, of Ongoing Alpha Oscillations
Authors: van Diepen Rosanne M. , Cohen Michael X , Denys Damiaan , Mazaheri Ali
Published: March 2015

The perception of near-threshold visual stimuli has been shown to depend in part on the phase (i.e., time in the cycle) of ongoing alpha (8–13 Hz) oscillations in the visual cortex relative to the onset of that stimulus. However, it is currently unknown whether the phase of the ongoing alpha activity can be manipulated by top–down factors such as attention or expectancy. Using three variants of a cross-modal attention paradigm with constant predictable stimulus onsets, we examined if cues signaling to attend to either the visual or the auditory domain influenced the phase of alpha oscillations in the associated sensory cortices. Importantly, intermixed in all three experiments, we included trials without a target to estimate the phase at target presentation without contamination from the early evoked responses. For these blank trials, at the time of expected target and distractor onset, we examined (1) the degree of the uniformity in phase angles across trials, (2) differences in phase angle uniformity compared with a pretarget baseline, and (3) phase angle differences between visual and auditory target conditions. Across all three experiments, we found that, although the cues induced a modulation in alpha power in occipital electrodes, neither the visual condition nor the auditory cue condition induced any significant phase-locking across trials during expected target or distractor presentation. These results suggest that, although alpha power can be modulated by top–down factors such as attention and expectation, the phase of the ongoing alpha oscillation is not under such control.
Oscillatory Theta Activity during Memory Formation and Its Impact on Overnight Consolidation: A Missing Link?
Authors: Heib Dominik P. J. , Hoedlmoser Kerstin , Anderer Peter , Gruber Georg , Zeitlhofer Josef , Schabus Manuel
Published: March 2015

Sleep has been shown to promote memory consolidation driven by certain oscillatory patterns, such as sleep spindles. However, sleep does not consolidate all newly encoded information uniformly but rather “selects” certain memories for consolidation. It is assumed that such selection depends on salience tags attached to the new memories before sleep. However, little is known about the underlying neuronal processes reflecting presleep memory tagging. The current study sought to address the question of whether event-related changes in spectral theta power (theta ERSP) during presleep memory formation could reflect memory tagging that influences subsequent consolidation during sleep. Twenty-four participants memorized 160 word pairs before sleep; in a separate laboratory visit, they performed a nonlearning control task. Memory performance was tested twice, directly before and after 8 hr of sleep. Results indicate that participants who improved their memory performance overnight displayed stronger theta ERSP during the memory task in comparison with the control task. They also displayed stronger memory task-related increases in fast sleep spindle activity. Furthermore, presleep theta activity was directly linked to fast sleep spindle activity, indicating that processes during memory formation might indeed reflect memory tagging that influences subsequent consolidation during sleep. Interestingly, our results further indicate that the suggested relation between sleep spindles and overnight performance change is not as direct as once believed. Rather, it appears to be mediated by processes beginning during presleep memory formation. We conclude that theta ERSP during presleep memory formation reflects cortico-hippocampal interactions that lead to a better long-term accessibility by tagging memories for sleep spindle-related reprocessing.
Syncitium in Motion: Exploring Movement in the Skeletal Muscle Cell Shared by Scientific and Artistic Nuclei
Authors: Hodgetts Stuart
Published: March 2015

The marriage of art and science often requires the sharing of unique characteristics. Skeletal muscle cells have provided a format in which the biology mimics the interaction of the artist and scientist within a common framework. This interaction, like the complex mechanism of fused muscle cells themselves, reveals and reminds those in both disciplines of the remarkable dynamic of movement between the two fields. This movement stimulates and rewards the artist and the scientist alike. For a scientist who works closely with artists, it is important to re-visit the fundamental concepts that drive the curiosity and stimulate the passion.
Re-purposing Life in an Anti-Disciplinary and Curiosity-Driven Context
Authors: Pelling Andrew E.
Published: March 2015

The earliest descriptions of the motions of living cells were marked by a sense of vitality. Indeed, these cellular motions have fascinated a diverse array of scientists and artists for centuries. The sustained interaction between artists and scientists in the author’s lab has led to an environment that fosters novel research directions and an appreciation of the intrinsic value of curiosity. Here, the author describes the anti-disciplinary research taking place in the lab and some of the discoveries that have revealed new paradigms governing the movement and behaviors of living cells in unnatural environments.
A Computationally Efficient Method for Incorporating Spike Waveform Information into Decoding Algorithms
Authors: Ventura Valérie , Todorova Sonia
Published: March 2015

Spike-based brain-computer interfaces (BCIs) have the potential to restore motor ability to people with paralysis and amputation, and have shown impressive performance in the lab. To transition BCI devices from the lab to the clinic, decoding must proceed automatically and in real time, which prohibits the use of algorithms that are computationally intensive or require manual tweaking. A common choice is to avoid spike sorting and treat the signal on each electrode as if it came from a single neuron, which is fast, easy, and therefore desirable for clinical use. But this approach ignores the kinematic information provided by individual neurons recorded on the same electrode. The contribution of this letter is a linear decoding model that extracts kinematic information from individual neurons without spike-sorting the electrode signals. The method relies on modeling sample averages of waveform features as functions of kinematics, which is automatic and requires minimal data storage and computation. In offline reconstruction of arm trajectories of a nonhuman primate performing reaching tasks, the proposed method performs as well as decoders based on expertly manually and automatically sorted spikes.
Surrogate Population Models for Large-Scale Neural Simulations
Authors: Tripp Bryan P.
Published: March 2015

Because different parts of the brain have rich interconnections, it is not possible to model small parts realistically in isolation. However, it is also impractical to simulate large neural systems in detail. This article outlines a new approach to multiscale modeling of neural systems that involves constructing efficient surrogate models of populations. Given a population of neuron models with correlated activity and with specific, nonrandom connections, a surrogate model is constructed in order to approximate the aggregate outputs of the population. The surrogate model requires less computation than the neural model, but it has a clear and specific relationship with the neural model. For example, approximate spike rasters for specific neurons can be derived from a simulation of the surrogate model. This article deals specifically with neural engineering framework (NEF) circuits of leaky-integrate-and-fire point neurons. Weighted sums of spikes are modeled by interpolating over latent variables in the population activity, and linear filters operate on gaussian random variables to approximate spike-related fluctuations. It is found that the surrogate models can often closely approximate network behavior with orders-of-magnitude reduction in computational demands, although there are certain systematic differences between the spiking and surrogate models. Since individual spikes are not modeled, some simulations can be performed with much longer steps sizes (e.g., 20 ms). Possible extensions to non-NEF networks and to more complex neuron models are discussed.
Molecular phylogenetics, systematics and host-plant associations of the Bruchidius albosparsus (Fåhraeus) species group (Coleoptera, Chrysomelidae, Bruchinae) with the description of four new species
Authors: DELOBEL ALEX , RU BRUNO LE , GENSON GWENAËLLE , MUSYOKA BOAZ K. , KERGOAT GAEL J.
Published: March 2015

Bruchidius Schilsky is a large paraphyletic genus of seed beetles (Coleoptera: Chrysomelidae: Bruchinae) which consists of multiple lineages that are usually associated with narrow sets of host-plants. In this study we focus on a group that mostly develops on wattle trees (acacias) belonging to the genus Vachellia Wight & Arn. This group originally included nine species and was designated as the Bruchidius centromaculatus (Allard) species group, but recent phylogenetic analyses revealed that these species belong to a much wider group of species with similar morphologies. For reasons of anteriority we call this enlarged group Bruchidius albosparsus (Fåhraeus). Here we review the morphology of species in this group and provide new diagnoses and ecological data for 10 species. The following combinations and synonymies are proposed: Bruchidius tanaensis (Pic, 1921) (= Bruchus tanaensis Pic, 1921) comb. nov. and Bruchidius albosparsus (Fåhraeus, 1839) (= Bruchus spadiceus Fåhraeus, 1839) syn. nov. Four new species are also described: B. eminingensis sp. nov., B. gerrardiicola sp. nov., B. glomeratus sp. nov. and B. haladai sp. nov. Finally we carried out molecular phylogenetic analyses on a multi-marker dataset of 59 specimens and 35 species, including 14 species from the group. The resulting trees allow us to confirm the monophyly of the group of interest and provide a more detailed picture of their evolutionary relationships.
Nanocladius (Plecopteracoluthus) shigaensis sp. nov. (Chironomidae: Orthocladiinae) whose larvae are phoretic on nymphs of stoneflies (Plecoptera) from Japan
Authors: INOUE YASUE , KOMORI CHIHARU , KOBAYASHI TADASHI , KONDO NATSUKO , UENO RYUHEI , TAKAMURA KENZI
Published: March 2015

We identified a new species, Nanocladius (Plecopteracoluthus) shigaensis, from Shiga and Gifu Prefectures, Japan, whose larvae are phoretic on nymphs of Plecoptera. Although this new species is morphologically similar to Nanocladius (Plecopteracoluthus) asiaticus Hayashi (1998), which is phoretic on Megaloptera larvae, it differs from N. (P.) asiaticus: the color of the larval head capsule is light brown in N. (P.) shigaensis and dark brown in N. (P.) asiaticus and the larval capsule index of the former is significantly larger than that of the latter. Moreover, analyses based on DNA sequence of cytochrome c oxidase subunit I (COI) supported the hypothesis that N. (P.) shigaensis and N. (P.) asiaticus are two distinct species. This is the first record of a phoretic chironomid on a plecopteran nymph in the Palaearctic region.
Spatial Patterns of Global Precipitation Change and Variability during 1901–2010
Authors: Gu Guojun , Adler Robert F.
Published: March 2015

This study examines global precipitation changes/variations during 1901–2010 by using the long-record GPCC land precipitation analysis, the NOAA/Cooperative Institute for Climate and Satellites (CICS) reconstructed (RECONS) precipitation analysis, and the CMIP5 outputs. In particular, spatial features of long-term precipitation changes and trends and decadal/interdecadal precipitation variations are explored by focusing on the effects of various physical mechanisms such as the anthropogenic greenhouse gas (GHG) and aerosol forcings and certain internal oscillations including the Pacific decadal variability (PDV) and Atlantic multidecadal oscillation (AMO).

Precipitation increases in the Northern Hemisphere (NH) mid- to high-latitude lands observed in GPCC can also be found in RECONS and model simulations. Over tropical/subtropical land areas, precipitation reductions generally appear in all products, but with large discrepancies on regional scales. Over ocean, consistent spatial structures of precipitation change also exist between RECONS and models. It is further found that these long-term changes/trends might be due to both anthropogenic GHG and aerosols. The aerosol effect estimated from CMIP5 historical simulations is then removed from the GPCC, RECONS, and AMIP simulations. These isolated GHG-related changes/trends have many similar spatial features when compared to the CMIP5 GHG-only simulations, especially in the zonal-mean context.

Both PDV and AMO have influence on spatial patterns of precipitation variations during the past century. In the NH middle to high latitudes, PDV and AMO have played an important role on interdecadal/multidecadal time scales. In several tropical/subtropical regions, their impacts may even become dominant for certain time spans including the recent past two decades. Therefore, these two internal mechanisms make the estimations of GHG and aerosol effects on precipitation on decadal/interdecadal time scales very challenging, especially on regional scales.
Quality Control of Antenna Alignment and Receiver Calibration Using the Sun: Adaptation to Midrange Weather Radar Observations at Low Elevation Angles
Authors: Altube Patricia , Bech Joan , Argemí Oriol , Rigo Tomeu
Published: March 2015

A quality control method for combined online monitoring of weather radar antenna pointing biases and receiver calibration using solar signals detected by an operational radar is adapted for application to midrange radar data (80–150 km). As the original method was developed using long-range data, additional criteria based on robust statistical estimators are imposed in the sun signature detection and selection process, allowing to discard observations biased by ground clutter or precipitation and to remove very influential outliers. The validity ranges of the physical model describing the solar interferences detected by the scanning radar antenna are explicitly defined and an equation for estimation of the effective scanning width in reception is provided in a thorough theoretical derivation. The method proposed reveals its sensitivity to changes in the antenna pointing accuracy and receiver calibration when applied to operational data obtained with three C-band radars during one year. A comparative study on the goodness of fit between a three- and a five-parameter model highlights the effect on the stability and accuracy of the antenna and receiver parameters retrieved for each radar system, considering the dissimilar information content of the observations collected by each radar. The performance of the proposed methodology under the effects of the presence of ground clutter and radio local area network interferences is discussed in the results presented.
Eosinophilic Mucin Otomastoiditis and Otopolyposis
Authors: Azadarmaki Roya , Westra William , Prasad Sanjay
Published: March 2015

Objective:

The purpose of this study is to introduce and define a disease entity on a continuum of eosinophilic otitis media: eosinophilic mucin otomastoiditis and otopolyposis.

Methods:

A case of a 66-year-old woman with complicated chronic otitis media is reported. A literature review of the National Library of Medicine’s online database, with a focus on eosinophilic otitis media and eosinophilic mucin rhinosinusitis, was performed.

Results:

The authors report the case of a 66-year-old woman with a history of asthma, chronic rhinosinusitis, nasal polyposis, and chronic otitis media who presented with allergic middle ear mucin and otic polyps. Treatment involved a tympanomastoidectomy with removal of otic polyps and steroid therapy.

Conclusion:

Eosinophilic mucin otomastoiditis with otopolyposis is a disease entity on a continuum of eosinophilic otitis media. This disease process shares similarities with eosinophilic mucin rhinosinusitis. Otic polypectomy and steroids are suggested therapeutic measures.
Revisiting Joseph Jockel's No Boundaries Upstairs
Authors: Heidt Daniel
Published: March 2015

This article reviews Joseph Jockel's classic study of bilateral Canadian–American defence arrangements during the early Cold War: No Boundaries Upstairs. It considers the political context in which Jockel wrote and highlights the bilateral experiences that contributed to his unique perspective. It also reviews Jockel's account and analysis of the evolution of Canadian–American continental defence cooperation from 1945 to 1958. Finally, this article describes the longevity of Jockel's analysis as well as how historians and political scientists continue to build on his research.
A Stress “Deafness” Effect in European Portuguese
Authors: Correia Susana , Butler Joseph , Vigário Marina , Frota Sónia
Published: March 2015

Research on the perception of word stress suggests that speakers of languages with non-predictable or variable stress (e.g., English and Spanish) are more efficient than speakers of languages with fixed stress (e.g., French and Finnish) at distinguishing nonsense words contrasting in stress location. In addition, segmental and suprasegmental cues to word stress may also impact on the ability of speakers to perceive stress. European Portuguese (EP) is a language with variable stress and vowel reduction. Previous studies on EP have identified duration as the main cue for stress. In the present study, we investigated the perception of word stress in EP, both in nuclear (NP) and post-nuclear (PN) positions, by means of three experiments. Experiment 1 was an ABX discrimination task with stress and phoneme contrasts, without vowel reduction. Experiments 2 and 3 were sequence recall tasks with stress and phoneme contrasts, vowel reduction being added to the stress contrast only in experiment 3. Results showed significantly higher error rates in the stress contrast condition than in the phoneme contrast condition, when duration alone (PN), or duration and pitch accents (NP), are present in the stimuli (experiments 1 and 2). When vowel reduction is added, EP speakers are able to perceive stress contrasts (experiment 3). The results show that vowel reduction appears to be the most robust cue for stress in EP. In the absence of vowel quality cues, a stress “deafness” effect may emerge in a language with non-predictable stress that combines both suprasegmental and segmental information to signal word stress. These findings have implications for claims of a prosodic-based cross-linguistic perception of word stress in the absence of vowel quality, and for stress “deafness” as a consequence of a predictable stress grammar.
Valuable Learning or “Spinning Their Wheels”? Understanding Excess Credits Earned by Community College Associate Degree Completers
Authors: Zeidenberg Matthew
Published: March 2015

Objective:

Excess credits earned by college students, over and above those required to complete their programs of study, have become increasingly a subject of interest and concern. There has been almost no research on the extent of these credits. This study focuses on all of the associate degree programs within one state’s community college system and measures the extent of excess credits within each program.

Method:

I created a measure of the number of excess credits earned relative to all credits earned and measured the extent to which colleges and programs vary in the levels of excess credits. Within particular academic programs, such as business, nursing, and general studies, I generated measures of the extent to which program-related and general education courses create excess credits. I examined the transcripts of some students who have earned excess credits, to see specifically what they did.

Results:

I take no normative position on the usefulness or harm of excess credits, but instead explore some of the factors that may generate them.

Contribution:

I suggest some policies that might be implemented to reduce excess credits, if desired.
Magnetic, fluorescent, and thermo-responsive poly(MMA-NIPAM-Tb(AA)3Phen)/Fe3O4 multifunctional nanospheres prepared by emulsifier-free emulsion polymerization
Authors: Gong Ying , Dai Jingwen , Li Huan , Wang Xin , Xiong Haoran , Zhang Quanyuan , Li Penghui , Yi Changfeng , Xu Zushun , Xu Haibo , Chu Paul K
Published: March 2015

Magnetic, luminescent, and thermoresponsive multifunctional nanospheres composed of modified Fe3O4 nanoparticles as the core and rare earth complex Tb(AA)3Phen as the shell are synthesized by emulsifier-free emulsion polymerization. The core–shell spherical structure has a size between 140 and 220 nm and exhibits strong green fluorescence of the rare earth complex Tb(AA)3Phen. In the R2 relaxivity and in vivo MRI studies, the R2 relaxivity of the nanospheres is 562.56 mM–1 s–1 and enhanced T2-weighted images are observed from the nanospheres in the liver and spleen after injection as a contrast agent. The excellent superparamagnetic, thermosensitive, and fluorescent properties render the nanospheres useful in biomedical engineering and optical imaging.
The Herpesvirus Proteases as Targets for Antiviral Chemotherapy
Authors: Waxman Lloyd , Darke Paul L
Published: March 2015

Viruses of the family Herpesviridae are responsible for a diverse set of human diseases. The available treatments are largely ineffective, with the exception of a few drugs for treatment of herpes simplex virus (HSV) infections. For several members of this DNA virus family, advances have been made recently in the biochemistry and structural biology of the essential viral protease, revealing common features that may be possible to exploit in the development of a new class of anti-herpesvirus agents. The herpesvirus proteases have been identified as belonging to a unique class of serine protease, with a Ser-His-His catalytic triad. A new, single domain protein fold has been determined by X-ray crystallography for the proteases of at least three different herpesviruses. Also unique for serine proteases, dimerization has been shown to be required for activity of the cytomegalovirus and HSV proteases. The dimerization requirement seriously impacts methods needed for productive, functional analysis and inhibitor discovery. The conserved functional and catalytic properties of the herpesvirus proteases lead to common considerations for this group of proteases in the early phases of inhibitor discovery. In general, classical serine protease inhibitors that react with active site residues do not readily inactivate the herpesvirus proteases. There has been progress however, with activated carbonyls that exploit the selective nucleophilicity of the active site serine. In addition, screening of chemical libraries has yielded novel structures as starting points for drug development. Recent crystal structures of the herpesvirus proteases now allow more direct interpretation of ligand structure—activity relationships. This review first describes basic functional aspects of herpesvirus protease biology and enzymology. Then we discuss inhibitors identified to date and the prospects for their future development.
Plant-Derived and Semi-Synthetic Calanolide Compounds with in Vitro Activity against Both Human Immunodeficiency Virus Type 1 and Human Cytomegalovirus
Authors: Xu Ze-Qi , Kern Earl R , Westbrook Louise , Allen Lois B , Buckheit Robert W , Tseng Christopher K-H , Jenta Tuah , Flavin Michael T
Published: March 2015

Plant-derived and semi-synthetic calanolide compounds with anti-human immunodeficiency virus type 1 (HIV-1) activity were tested for anti-human cytomegalovirus (HCMV) activity in both cytopathic effect inhibition and plaque reduction assays. The results indicated that the anti-HCMV activity of calanolide compounds does not correlate with their activity against HIV-1. The semi-synthetic 12-keto derivatives tended to be more active against HCMV than the corresponding 12-OH congeners, which were more active against HIV-1. It appeared that the 7,8-unsaturated double bond in the chromene ring played a certain role in maintaining activities against both HCMV and HIV-1. Saturation of the double bond increased the EC50 values against both viruses, with concomitant increase in toxicity. The calanolide compounds reported here are the first non-nucleoside analogues capable of inhibiting both HIV-1 and HCMV and, therefore, may be useful chemoprophylactic agents for HCMV in HIV-infected people or vice versa.
Importance of the Alanine Methyl Ester Side Chain for the Biological Activity Profile of Dual-Function Phenyl Phosphate Derivatives of Bromo-Methoxy-Zidovudine
Authors: Venkatachalam TK , D'Cruz OJ , Uckun FM
Published: March 2015

In a systematic search for developing a virucidal spermicide with potent anti-human immunodeficiency virus (HIV) and spermicidal activities, we synthesized and evaluated 14 phosphoramidate derivatives of 5-bromo-6-methoxy-zidovu-dine (PP-BMZ) with differing amino acid ester side chains and para substitutions on the phenyl moiety. Anti-HIV activity was tested by measuring viral p24 antigen production as a marker of viral replication in HIV-1-infected human peripheral blood mononuclear cells. The effect of various PP-BMZ compounds on human sperm motion kinematics was analysed by computer-assisted sperm analysis. Varying the Ala side chain of the phosphoramidate group to other non-polar amino acids, including the cyclic amino acids proline and tryptophan, led to significant alterations in both anti-HIV and spermicidal activities. Our findings highlight the necessity of the Ala side chain and the presence of an electron-withdrawing para-bromo substi-tutent on the phenyl moiety in addition to the bromo-methoxy functional groups on the thymine ring for the PP-BMZ compounds to be effective virucidal spermicides. These membrane permeable dual-function nucleoside analogues may provide the basis for a new strategy aimed at prevention of the sexual transmission of HIV while providing fertility control for women.
Thienothiadiazine 2,2-Dioxide Acyclonucleosides: Synthesis and Antiviral Activity
Authors: Martinez A , Esteban AI , Gil C , Conde S , Andrei G , Snoeck R , Balzarini J , De Clercq E
Published: March 2015

The synthesis of acyclonucleosides derived from thieno[3,2-c] and thieno[2,3-c][1,2,6]thiadiazine 2,2-dioxides was achieved following the silylation method. Lipase-mediated methodology was employed for deprotection of the acyclic moieties. The antiviral effects were determined against a broad spectrum of viruses, including cytomegalovirus (CMV) and varicella zoster virus (VZV). Only minor antiviral activity against VZV was observed for those acyclonucleosides carrying a benzyl group.
Antiviral and Virucidal Activities against Arenaviruses of Zinc-Finger Active Compounds
Authors: Garcia CC , Candurra NA , Damonte EB
Published: March 2015

Fifteen antiretroviral Zn-finger active compounds with diverse chemical structures, including azoic compounds, hydrazide derivatives, disulphide-based reagents and others were screened in vitro against Junin virus (JUNV), the aetiological agent of Argentine haemorrhagic fever, by a virus yield inhibition assay in Vero cells. Cytotoxicity was evaluated simultaneously by the MTT method. Of the compounds, three were totally inactive as antivirals, nine presented moderate anti JUNV-activity and three were truly active with EC50 (effective concentration 50%) values in the range 6.5–9.3 μM and with selectivity indices greater than 10. The most active inhibitors, named NSC20625, 3–7 and 2–71, demonstrated a broad range of action against arenaviruses, including several attenuated and pathogenic strains of JUNV as well as the antigenically related Tacaribe virus (TACV) and Pichinde virus (PICV). The direct treatment of JUNV and TACV virions with the compounds showed two types of behaviour: the aromatic disulphide NSC20625 was a very potent virucidal agent, whereas the other two compounds exhibited moderate or negligible virus-inactivating properties.
Anti-Varicella—Zoster Virus Bicyclic Nucleosides: Replacement of Furo by Pyrro Base Reduces Antiviral Potency
Authors: McGuigan C , Pathirana RN , Jones G , Andrei G , Snoeck R , De Clercq E , Balzarini J
Published: March 2015

We have recently reported the discovery of an entirely new category of potent antiviral agents based on novel deoxynucleoside analogues with unusual fluorescent bicyclic furo base moieties. To probe structure-activity relationships and to seek to optimize the properties of the lead compounds, we prepared pyrro analogues of the furo systems. We herein report the synthesis, characterization and antiviral evaluation of these novel compounds.
Correlation between Intracellular Pharmacological Activation of Nucleoside Analogues and HIV Suppression in Vitro
Authors: Hoggard PG , Sales SD , Kewn S , Sunderland D , Khoo SH , Hart CA , Back DJ
Published: March 2015

Following intracellular activation of HIV nucleoside analogue reverse transcriptase inhibitors, their triphosphates (ddNTPs) compete with endogenous nucleoside triphosphates (dNTPs) for incorporation into proviral DNA. In this study we have examined the effect of combinations of two thymidine analogues, stavudine (d4T) and zidovudine (ZDV), and two cytidine analogues, lamivudine (3TC) and zalcitabine (ddC) on intracellular drug activation and on the relevant competing dNTP in uninfected and persistently HIV-infected cells. Endogenous triphosphates of deoxycytidine (dCTP) and deoxythymidine (dTTP) were measured using a template primer assay and the ratio of ddNTP:dNTP was calculated. Antiviral activity of two-drug combinations was also assayed by p24 ELISA. A significant reduction in d4T triphosphate (d4TTP) [0.11±0.09 pmol/106 cells to undetectable (<0.01); P=0.039] in the presence of equimolar concentrations of ZDV and d4T, resulted in a decrease in the d4TTP/dTTP ratio of 90%. ZDVTP/dTTP was not significantly altered in the presence of d4T. 3TC (10 μM) reduced total ddC phosphates by 57% and ddCTP/dCTP by 27%. 3TC phosphorylation was comparatively unaffected by ddC, up to a concentration of 10 μ ddC (>100 times the plasma concentration achieved following standard dosing). 3TC plus ddC resulted in greater p24 inhibition than 3TC or ddC alone (P<0.001). Combining one thymidine analogue (ZDV or d4T) with one cytidine analogue (3TC or ddC) resulted in greater inhibition of p24 inhibition than with any single agent. From a pharmacological viewpoint, the combination of ZDV plus d4T should be avoided, but in vitro the combination of 3TC plus ddC confers modest benefit over either drug alone. This in vitro study illustrates that decreases in ddNTP/dNTP are consistent with a reduction in antiviral effect.
Resistance of Hepatitis B Virus to Antiviral Drugs: Current Aspects and Directions for Future Investigation
Authors: Delaney William E , Locarnini Stephen , Shaw Tim
Published: March 2015

Despite the existence of vaccines, chronic hepatitis B virus (HBV) infection remains a major health problem worldwide. Interferon therapy successfully controls infection in only a small percentage of chronically infected individuals. The recent approval of the nucleoside analogue lamivudine for the treatment of chronic HBV infection has ushered in a new era of antiviral therapy. While lamivudine is highly effective at controlling viral infection short-term, prolonged therapy has been associated with an increasing incidence of viral resistance. Thus, it appears that lamivudine alone will not be sufficient to control chronic viral infection in the majority of individuals. In addition to lamivudine, several new nucleoside and nucleotide analogues that show promising antihepadnaviral activity are in various stages of development. Lamivudine resistance has been found to confer cross-resistance to some of these compounds and it is likely that resistance to newer antivirals may also develop during prolonged use. Drug resistance therefore poses a major threat to nucleoside analogue-based therapies for chronic HBV infection. Fortunately, combination chemotherapy (antiviral therapy with two or more agents) can minimize the chance that resistance will develop and can be expected to achieve sustained reductions in viral load, provided that suitable combinations of agents are chosen. Here we review the basis of drug resistance in HBV, with emphasis on aspects that are likely to affect drug choice in future.
Inhibition of HIV-1 Replication in Macrophages by Red Blood Cell-Mediated Delivery of a Heterodinucleotide of Azidothymidine and 9-(R)-2-(Phosphono Methoxypropyl)adenine
Authors: Franchetti P , Rossi L , Cappellacci L , Pasqualini M , Grifantini M , Balestra E , Forbici F , Perno C-F , Serafini S , Magnani M
Published: March 2015

Monocyte-derived macrophages (M/M) are considered important in vivo reservoirs for different kinds of viruses, including HIV. Hence, therapeutic strategies are urgently needed to protect these cells from virus infection or to control viral replication. In this paper, we report the synthesis, target delivery and in vitro efficacy of a new heterodinucleotide (AZTpPMPA), able to inhibit HIV-1 production in human macrophages. AZTpPMPA consists of two established anti-HIV drugs [zidovudine (AZT) and tenofovir (PMPA)] chemically coupled together by a phosphate bridge. This drug is not able to prevent p24 production when administered for 18 h to M/M experimentally infected with HIV-1 Bal (inhibition 27%), but can almost completely suppress virus production when given encapsulated into autologous erythrocytes (inhibition of p24 production 97%). AZTpPMPA is slowly converted to PMPA, AZT monophosphate and AZT (36 h half-life at 37°C) by cell-resident enzymes. Thus AZTpPMPA should be considered a new prodrug of AZT and PMPA that is able to provide stechiometric amounts of both nucleoside analogues to macrophage cells and to overcome the low phosphorylating activity of M/M for AZT and the modest permeability of PMPA.
Synthesis, Biological Activity, Pharmacokinetic Properties and Molecular Modelling Studies of Novel 1H,3H-Oxazolo[3,4-a]Benzimidazoles: Non-Nucleoside HIV-1 Reverse Transcriptase Inhibitors
Authors: Chimirri A , Monforte P , Rao A , Zappalà M , Monforte AM , De Sarro G , Pannecouque C , Witvrouw M , Balzarini J , De Clercq E
Published: March 2015

New 1H,3H-oxazolo[3,4-a]benzimidazoles (OBZs) were synthesized as HIV-1 non-nucleoside reverse transcriptase inhibitors (NNRTI) to extend the structure-activity relationships observed for an early series of related 1H,3H-thiazolo[3,4-a]benzimidazole derivatives (TBZs). The new compounds showed inhibitory activity against the replication of various HIV-1 strains, including NNRTI-resistant strains. Testing of a representative OBZ derivative in an HPLC assay on biological fluids, indicated that the sulphur substitution appreciably improved the metabolic stability of the TBZ compound. In addition, molecular modelling studies demonstrated that OBZs, TBZs and other NNRTIs have similar structural properties, that is a butterfly-like conformation, which is a key structural requirement for reverse transcriptase inhibition.
Iminosugars as Possible Broad Spectrum anti Hepatitis Virus Agents: The Glucovirs and Alkovirs
Authors: Block Timothy M , Jordan Robert
Published: March 2015

Glucosidases in the endoplasmic reticulum (ER) mediate the first step in processing N-linked oligosaccharides. Recent evidence suggests that morphogenesis and secretion of members of the hepatitis B and flavivirus families are more dependent on these enzymes than are most host glycoproteins. Thus, it is possible that glucosidase inhibitors can be designed that are safe and selective for the treatment of hepatitis B and possibly C (since hepatitis C virus is a member of the flavivirus family), making them broad spectrum with respect to hepatitis viruses. Numerous pharmacological and genetic dissections support the notion that glucosidase inhibition can have an antiviral effect, and imino sugars that competitively inhibit ER glucosidases have been proposed as anti-hepatitis drug candidates. We call this family of compounds ‘glucovirs′. Recently, however, alkylated imino sugars that retain substantial antiviral activity but lack glucosidase inhibitory activity have been described. These compounds are called ‘alkovirs' and their mechanism of action is unknown. This review considers the rationale of the glucovir and alkovir approach to the treatment of hepatitis B and C.
In vitro Activity of Polyhydroxycarboxylates against Herpesviruses and Hiv
Authors: Meerbach Astrid , Neyts Johan , Balzarini Jan , Helbig Björn , De Clercq Erik , Wutzler Peter
Published: March 2015

The antiviral activity of 17 polyhydroxycarboxylates derived from phenolic compounds was evaluated against herpesviruses and HIV. When present during virus adsorption several of the polymers exhibited potent activity against herpes simplex virus type 1 (HSV-1), HSV-2, thymidine kinase deficient HSV-1, human cytomegalovirus (HCMV) and HIV-1 and HIV-2 at concentrations that were not toxic to the host cells. A close correlation was found between the 50% inhibitory concentrations of the polyhydroxycarboxylates against HCMV-induced cytopathicity, their inhibitory effect on the expression of HCMV-specific immediate early antigens and their inhibitory effects on HCMV adsorption to the cells. The antiviral activity of the phenolic polymers was dependent on the presence of a sufficient number of carboxylic groups. The mechanism of antiviral action of the polyhydroxycarboxylates can thus be ascribed to inhibition of virus adsorption. This type of compound may have potential in a vaginal gel to prevent sexual transmission of HSV and HIV.
Inhibitory Effect of Modified 5′-Capped Short RNA Fragments on Influenza Virus RNA Polymerase Gene Expression
Authors: Tado Motoki , Abe Takayuki , Hatta Toshifumi , Ishikawa Masahide , Nakada Susumu , Yokota Tomoyuki , Takaku Hiroshi
Published: March 2015

We have shown previously that the 5′-capped short phosphodiester RNA fragments, Cap decoy, (Gm 12 nt) are potent inhibitors of influenza virus RNA polymerase gene expression. Here we investigate the modified capped RNA derivative containing phosphorothioate oligonucleotides (Cap decoy) as a potential influenza virus RNA polymerase inhibitor. The modified 5′-capped short phosphorothioate RNA fragments (Gms 12–15 nt) with the 5′-capped structure (m7GpppGm) were synthesized by T7 RNA polymerase. The 5′-capped short RNA fragments (Gms 12–15 nt) were encapsulated in liposome particulates and tested for their inhibitory effects on influenza virus RNA polymerase gene expression in the clone 76 cells. The 12–15 nt long Gms RNA fragments showed highly inhibitory effects. By contrast, the inhibitory effects of the 13 nt long short RNA fragments (Gm 13 nt) were considerably less in comparison with the 5′-capped short phosphorothioate RNA fragments (Gms 12–15 nt). In particular, the various Gms RNA chain lengths showed no significant differences in the inhibition of influenza virus RNA polymerase gene expression. Furthermore, the capped RNA with a phosphorothioate backbone was resistant to nuclease activity. These phosphorothioate RNA fragments exhibited higher inhibitory activity than the 5′-capped short RNA fragments (Gm 12 nt). These decoys may prove to be useful in anti-influenza virus therapeutics.
In Search of Authentic Inhibitors of HIV-1 Integration
Authors: Debyser Zeger , Cherepanov Peter , Van Maele Bénédicte , De Clercq Erik , Witvrouw Myriam
Published: March 2015

Current strategies for the treatment of HIV infection are based on cocktails of drugs that target the viral reverse transcriptase or protease enzymes. At present, the clinical benefit of this combination therapy for HIV-infected patients is considerable, although it is not clear how long this effect will last taking into account the emergence of multiple drug-resistant viral strains. Addition of new anti-HIV drugs targeting additional steps of the viral replication cycle may increase the potency of inhibition and prevent resistance development. During HIV replication, integration of the viral genome into the cellular chromosome is an essential step catalysed by the viral integrase. Although HIV integrase is an attractive target for antiviral therapy, so far all research efforts have led to the identification of only one series of compounds that selectively inhibit the integration step during HIV replication, namely the diketo acids. In this review we try to address the question why it has proven so difficult to find potent and selective integrase inhibitors. We point to potential pitfalls in defining an inhibitor as an authentic integrase inhibitor, and propose new strategies and technologies for the discovery of authentic HIV integration inhibitors.
A Combination of Poor Adherence and a Low Baseline Susceptibility Score is Highly Predictive for HAART Failure
Authors: Van Vaerenbergh Kristien , De Geest Sabina , Derdelinckx Inge , Bobbaers Herman , Carbonez An , Deschamps Ann , De Graeve Veerle , De Saar Veerle , Ceunen Helga , De Smet Koen , Maes Bart , Peetermans Willy , Schrooten Yoeri , Desmyter Jan , De Clercq Erik , Van Ranst Marc , Van Wijngaerden Eric , Vandamme Anne-Mieke
Published: March 2015

The relationship between adherence, virological response to highly active antiretroviral therapy (HAART) and the presence and development of genotypic resistance was assessed in 41 HIV-infected patients on HAART. Four adherence parameters (drug taking adherence, dosing adherence, timing adherence and drug holidays) were scored prospectively using electronic event monitoring. Genotypic resistance at baseline and after therapy failure was scored retrospectively and a genotype-based susceptibility score was calculated. Overall median adherence rates were high. All adherence parameters were better in virological responders ( n=31) compared to non-responders ( n=10), drug taking adherence and number of drug holidays being significantly different. Responders had a significantly higher susceptibility score. Stepwise logistic regression showed that the number of drug holidays and a low susceptibility score were highly predictive for therapy failure. Despite the presence of a limited number of baseline resistance mutations, perfectly adherent patients can control virus replication for a prolonged period.
Short-Term Treatment with Novel Ribonucleotide Reductase Inhibitors Trimidox and Didox Reverses Late-Stage Murine Retrovirus-Induced Lymphoproliferative Disease with Less Bone Marrow Toxicity Than Hydroxyurea
Authors: Mayhew CN , Phillips JD , Cibull ML , Elford HL , Gallicchio VS
Published: March 2015

We evaluated the ability of a short course of treatment with the ribonucleotide reductase (RR) inhibitor hydroxyurea (HU) and two novel RR inhibitors Trimidox (TX) and Didox (DX) to influence late-stage murine retrovirus-induced lymphoproliferative disease. LPBM5 murine leukaemia virus retrovirus-infected mice were treated daily with HU, TX or DX for 4 weeks, beginning 9 weeks post-infection, after development of immunodeficiency and lymphoproliferative disease. Drug effects on disease progression were determined by evaluating spleen weight and histology. Effects on haematopoiesis were determined by measuring peripheral blood indices (white blood cells and haematocrit) and assay of femur cellularity and femoral and splenic content of colony-forming units granulocytemacrophage (CFU-GM) and burst-forming units-erythroid (BFU-E). HU, TX and DX partially reversed late-stage retrovirus-induced disease, resulting in spleen weights significantly below pre-treatment values. Spleen histology was also improved by RR inhibitor treatment (DX>TX>HU). However, as expected, HU was significantly myelosuppressive, inducing a reduction in peripheral indices associated with depletion of femoral CFU-GM and BFU-E. In contrast, although TX and DX were moderately myelosuppressive, both drugs were significantly better tolerated than HU. In summary, short-term treatment in late-stage murine retroviral disease with HU, TX or DX induced dramatic reversal of disease pathophysiology. However, the novel RR inhibitors TX and DX had more effective activity and significantly less bone marrow toxicity than HU.
Evidence for Dual Effects of DNA-Reactive Bile Acid Derivatives (Bamets) on Hepatitis B Virus Life Cycle in an In Vitro Replicative System
Authors: Romero Marta R , Martinez-Diez Maria C , Larena Monica G , MacIas Rocio IR , Dominguez Mercedes , Garcia-Monzon Carmelo , Serrano Maria A , Marin Jose JG
Published: March 2015

A liver targeting strategy to direct antiviral drugs toward hepatitis B virus (HBV) was investigated. As model drugs we used cisplatin-bile acid derivatives (Bamets) to determine the production of virions by HBV-transfected hepatoblastoma cells (HepG2 2.2.15). Drug uptake was determined using flameless atomic absorption spectrometry to measure platinum cell contents. Cytotoxic effect was determined by formazan formation and neutral red uptake tests. The release of viral surface protein was evaluated by ELISA. The abundance of HBV-DNA in the medium was determined by quantitative real-time PCR and its structure by Southern blot analysis. The uptake of Bamets by HepG2 2.2.15 cells was higher than that of cisplatin. At concentrations lower than 10 μM, distinct Bamets have no toxic effect on host cells, whereas cisplatin dramatically reduced cell viability at concentrations higher than 1 μM. All the drugs tested inhibited the release of viral proteins to the medium, but induced a marked and progressive dose-dependent increase in the amount of viral DNA in the medium. This was mainly due to the release of short fragments of HBV-DNA in the case of cisplatin. On the contrary, Bamets induced an enhanced release of circular forms of HBV-DNA. These findings suggest the existence of a dual effect of Bamets on HBV life-cycle by enhancing the production of DNA replicative intermediates but reducing the secretion of complete virions. Altogether these characteristics recommend consideration of these compounds as a useful experimental tool in the investigation of novel liver targeted therapeutic agents based on bile acid derivatives for the treatment of HBV infections, or to carry out further studies on the HBV life cycle.
In Vitro Anti-Influenza Virus Activity of the Pavine Alkaloid (-)-Thalimonine Isolated from Thalictrum Simplex L
Authors: Serkedjieva Julia , Velcheva Maria
Published: March 2015

The pavine alkaloid (-)-thalimonine (Thl), isolated from the Mongolian plant Thalictrum simplex inhibited markedly the reproduction of influenza virus A/Germany/27, str. Weybridge (H7N7) and A/Germany/34, str. Rostock (H7N1) in cell cultures of chicken embryo fibroblasts. In a number of assays at a non-toxic concentration range of 0.1–6.4 μM the alkaloid inhibited viral reproduction in a selective and specific way (selectivity index=640, 106.6, respectively). Expression of viral glycoproteins haemagglutinin (HA), neuraminidase (NA) and nucleoprotein (NP) on the surface of infected cells, virus-induced cytopathic effect, infectious virus yields, HA production and virus-specific protein synthesis were all reduced. The inhibition was dose-related and depended on virus inoculum. The time of addition experiments indicated that viral reproduction was markedly inhibited when Thl was added at 4–5 h of infection. No inactivating effect on extracellular virus was found.