bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between vascular function indexes, renal arteriolosclerosis, and renal clinical outcomes in chronic kidney disease
Authors: Namikoshi Tamehachi , Fujimoto Sohachi , Yorimitsu Daisuke , Ihoriya Chieko , Fujimoto Yasuo , Komai Norio , Sasaki Tamaki , Kashihara Naoki
Published: April 2015

Aim

Hypertension contributes critically to the development of renal arteriolosclerosis in chronic kidney disease (CKD), but the impact of vascular function indexes including central blood pressure on renal arteriolosclerosis has not been investigated. We determined whether vascular function indexes were related to renal arteriolosclerosis and renal clinical outcomes in CKD.

Methods

This cross‐sectional study was implemented in our hospital. Subjects were in‐patients with CKD aged ≥20 years who underwent a renal biopsy. Vascular function indexes included central systolic blood pressure (SBP), cardio–ankle vascular index (CAVI), and renal resistive index. Central SBP was measured non‐invasively using an automated device. Arteriolosclerosis was assessed histologically. Renal clinical outcomes included estimated glomerular filtration rate using serum creatinine (eGFRcreat) or cystatin C (eGFRcys), and the urinary albumin–creatinine ratio.

Results

Among vascular function indexes, central SBP was weakly correlated with renal arteriolosclerosis (n = 55). Renal arteriolosclerosis was increased in hypertensive or hyperuricaemic patients, and negatively correlated with serum high‐density lipoprotein (HDL) cholesterol and eGFRcys, which were independent risk factors for renal arteriolosclerosis in a stepwise multivariate regression analysis. Of the vascular function indexes, CAVI showed the strongest correlation with all renal clinical outcomes. Central SBP was correlated with only urinary albumin–creatinine ratio, while renal resistive index was correlated with eGFRcreat and urinary albumin–creatinine ratio.

Conclusion

Decreased serum HDL cholesterol was independently and most closely associated with renal arteriolosclerosis. Of the vascular function indexes, CAVI had the greatest impact on renal clinical outcomes, although it was not associated with renal arteriolosclerosis.
Oakleaf: an S locus‐linked mutation of Primula vulgaris that affects leaf and flower development
Authors: Cocker Jonathan M. , Webster Margaret A. , Li Jinhong , Wright Jonathan , Kaithakottil Gemy , Swarbreck David , Gilmartin Philip M.
Published: April 2015

In Primula vulgaris outcrossing is promoted through reciprocal herkogamy with insect‐mediated cross‐pollination between pin and thrum form flowers. Development of heteromorphic flowers is coordinated by genes at the S locus. To underpin construction of a genetic map facilitating isolation of these S locus genes, we have characterised Oakleaf, a novel S locus‐linked mutant phenotype.

We combine phenotypic observation of flower and leaf development, with classical genetic analysis and next‐generation sequencing to address the molecular basis of Oakleaf.

Oakleaf is a dominant mutation that affects both leaf and flower development; plants produce distinctive lobed leaves, with occasional ectopic meristems on the veins. This phenotype is reminiscent of overexpression of Class I KNOX‐homeodomain transcription factors. We describe the structure and expression of all eight P. vulgaris PvKNOX genes in both wild‐type and Oakleaf plants, and present comparative transcriptome analysis of leaves and flowers from Oakleaf and wild‐type plants.

Oakleaf provides a new phenotypic marker for genetic analysis of the Primula S locus. We show that none of the Class I PvKNOX genes are strongly upregulated in Oakleaf leaves and flowers, and identify cohorts of 507 upregulated and 314 downregulated genes in the Oakleaf mutant.
Distinct sensitivities to phosphate deprivation suggest that RGF peptides play disparate roles in Arabidopsis thaliana root development
Authors: Cederholm Heidi M. , Benfey Philip N.
Published: April 2015

Growing agricultural demands in the face of impending inorganic phosphate (Pi) shortages underscore a need for a better understanding of plant development under conditions of Pi deprivation. Pi is an essential nutrient that is a major component of fertilizer. Plants have evolved strategies to improve the acquisition of this nutrient by altering root development under shortage conditions. We show that signaling peptides thought to act redundantly in Arabidopsis thaliana development have distinct functions in response to Pi deprivation.

Using microscopy and confocal imaging, roots were analyzed for growth rate and cellular composition. Using expression microarrays, genes influencing development in response to phosphate deprivation were identified.

ROOT GROWTH FACTOR1 (RGF1) and RGF2 influenced different aspects of root development under conditions of Pi deprivation. We found that RGF2 influenced the longitudinal growth rate in the primary root in response to Pi deprivation, whereas RGF1 affected circumferential cell number in the root meristem.

These data suggest that the mechanisms controlling adaptive development can depend on disparate functions of genes thought to act redundantly, thus elucidating new functions for important developmental regulators.
External guide sequence technology: a path to development of novel antimicrobial therapeutics
Authors: Davies‐Sala Carol , Soler‐Bistué Alfonso , Bonomo Robert A. , Zorreguieta Angeles , Tolmasky Marcelo E.
Published: April 2015

RNase P is a ribozyme originally identified for its role in maturation of tRNAs by cleavage of precursor tRNAs (pre‐tRNAs) at the 5′‐end termini. RNase P is a ribonucleoprotein consisting of a catalytic RNA molecule and, depending on the organism, one or more cofactor proteins. The site of cleavage of a pre‐tRNA is identified by its tertiary structure; and any RNA molecule can be cleaved by RNase P as long as the RNA forms a duplex that resembles the regional structure in the pre‐tRNA. When the antisense sequence that forms the duplex with the strand that is subsequently cleaved by RNase P is in a separate molecule, it is called an external guide sequence (EGS). These fundamental observations are the basis for EGS technology, which consists of inhibiting gene expression by utilizing an EGS that elicits RNase P–mediated cleavage of a target mRNA molecule. EGS technology has been used to inhibit expression of a wide variety of genes, and may help development of novel treatments of diseases, including multidrug‐resistant bacterial and viral infections.
Evaluation of resting state gamma power as a response marker in schizophrenia
Authors: Mitra Sayantanava , Nizamie S. Haque , Goyal Nishant , Tikka Sai Krishna
Published: April 2015

Aims

An abnormal activity in the electroencephalography (EEG) gamma band (>30 Hz) has been demonstrated in schizophrenia and this has been suggested to be reflecting a deficit in the development and maturation of the basic cognitive functions of attention, working memory and sensory processing. Hypothesizing gamma oscillatory activity as a potential EEG biomarker to antipsychotic response in schizophrenia, the present study aimed at measuring baseline spontaneous gamma activity in schizophrenia patients, and evaluating its response to antipsychotic treatment over 8 weeks.

Methods

Fifteen drug‐free/naïve patients were recruited, compared at baseline with 15 age‐, sex‐ and education‐matched healthy controls, and were followed up for 8 weeks' treatment on antipsychotics. Resting state EEG waves were recorded using high (192‐channel) resolution EEG at admission, 4 weeks and 8 weeks. Spectral power was calculated using fast Fourier transformation, Hanning window. The power was averaged region‐wise over nine regions in three frequency ranges (30–50 Hz, 50–70 Hz, 70–100 Hz).

Results

Patients and controls differed significantly at intake in terms of left temporal and parietal high (70–100 Hz) gamma power. Consequently, no significant differences were seen over the course of antipsychotic treatment on gamma spectral power in any of the regions.

Conclusions

Lack of significant effect of treatment on gamma power suggests that these gamma oscillations may be trait markers in schizophrenia.
Opportunities of marker‐assisted selection for rice fragrance through marker–trait association analysis of microsatellites and gene‐based markers
Authors: Golestan Hashemi F. S. , Rafii M. Y. , Razi Ismail M. , Mohamed M. T. M. , Rahim H. A. , Latif M. A. , Aslani F.
Published: April 2015

Developing fragrant rice through marker‐assisted/aided selection (MAS) is an economical and profitable approach worldwide for the enrichment of an elite genetic background with a pleasant aroma. The PCR‐based DNA markers that distinguish the alleles of major fragrance genes in rice have been synthesised to develop rice scent biofortification through MAS. Thus, the present study examined the aroma biofortification potential of these co‐dominant markers in a germplasm panel of 189 F2 progeny developed from crosses between a non‐aromatic variety (MR84) and a highly aromatic but low‐yielding variety (MRQ74) to determine the most influential diagnostic markers for fragrance biofortification. The SSRs and functional DNA markers RM5633 (on chromosome 4), RM515, RM223, L06, NKSbad2, FMbadh2‐E7, BADEX7‐5, Aro7 and SCU015RM (on chromosome 8) were highly associated with the 2AP (2‐acetyl‐1‐pyrroline) content across the population. The alleles traced via these markers were also in high linkage disequilibrium (R2 > 0.70) and explained approximately 12.1, 27.05, 27.05, 27.05, 25.42, 25.42, 20.53, 20.43 and 20.18% of the total phenotypic variation observed for these biomarkers, respectively. F2 plants harbouring the favourable alleles of these effective markers produced higher levels of fragrance. Hence, these rice plants can be used as donor parents to increase the development of fragrance‐biofortified tropical rice varieties adapted to growing conditions and consumer preferences, thus contributing to the global rice market.
Failure of temporal selectivity: Electrophysiological evidence for (mis)selection of distractors during the attentional blink
Authors: Bourassa Marie‐Ève , Vachon François , Brisson Benoit
Published: April 2015

The attentional blink (AB) refers to the impairment in accurate report of a second target (T2) when presented shortly after a first target (T1) in a rapid serial visual presentation of distractors. The goal of the present study was to determine whether the AB is caused by a failure in early selection processes, which leads to the selection and consolidation of the wrong item in working memory, by measuring the frequency‐related P3 to T2 and to T2 + 1. During the AB, an attenuation of the P3 to T2 was observed, as well as an increase in the amplitude of the P3 to T2 + 1. Whereas the P3 to T2 was observed only when T2 was correctly reported, the P3 to T2 + 1 was observed only in trials where T2 was incorrectly reported, and its amplitude was correlated to individual differences in misselection rate. These results support the claim that failure of temporal selection underlies the AB.
Effectiveness of a psychoeducational programme for informal caregivers of older adults
Authors: Alves Sara , Teixeira Laetitia , Azevedo Maria J. , Duarte Mafalda , Paúl Constança
Published: April 2015

Purpose of the study

This study explored the effectiveness of psychoeducational programme for informal caregivers called ‘Caring at home’.

Design and methods

The sample included 60 informal caregivers of elderly people (≥60 years) living in the community. Socio‐demographic information of informal caregivers and care receivers was collected with the COPE Index. The assessment protocol comprised four scales: 12‐item Short‐Form Health Survey, General Health Questionnaire‐12, Positive Aspects of Caregiving and Caregiver Strain Index Modified. There were five assessment moments: a pretest, a post‐test and three follow‐ups.

Results

Informal caregivers had an average age of 52.08 years (SD = 9.11) and were mainly women (90.0%), married/partnered (75.0%) and children of the care recipients (61.7%). The median time spent in care was 12 hours/day (IQR = 18). Results showed a favourable evolution after the psychoeducational intervention, a significant improvement in caregivers' mental health (p = 0.03) from pretest to 1st follow‐up (2/3 months after the psychoeducational intervention) and the maintenance of the other assessed dimensions of the caregivers' behaviour.

Implications

The psychoeducational intervention seems to promote improvements in mental health and the maintenance of other favourable conditions at baseline. These results may indicate that psychoeducational programme are beneficial to minimise or prevent adverse effects of caregiving.
Management of Polypharmacy in Dialysis Patients
Authors: St. Peter Wendy L.
Published: April 2015

Most patients receiving dialysis have other common chronic conditions in addition to end‐stage renal disease, including hypertension, diabetes, cardiovascular disease, and mineral and bone disorder, all of which require long‐term medication management. Dialysis patients take an average of 10–12 prescribed and over‐the‐counter medications from an average of 4.7 prescribers, and an average of 19 pills per day. Thus, reducing polypharmacy is not adequate as a medication therapy goal for these patients. Instead, the dialysis community should focus on ensuring that all patients receive medications that are appropriate, effective, safe and convenient. Barriers to this include a fragmented health care system with inadequate communication between multiple prescribers and pharmacies, and frequent care transitions between ambulatory care sites (dialysis centre, ambulatory primary care practice, ambulatory specialty practice) and the hospital, skilled nursing facility or long‐term care facility. Three distinct processes are necessary to prevent and solve the resultant medication‐related problems (and reduce polypharmacy). These are medication reconciliation (creating an accurate medication list that reflects all medications the patients is taking and how they are being taken), medication review (evaluating the list for appropriateness, effectiveness, safety and convenience in conjunction with the patient's health status), and ongoing patient‐centred medication therapy management (e.g., developing treatment plans centred on each patient's medication‐related goals). A team approach including pharmacists as part of the dialysis team with the dialysis facility as the primary medication home is needed.
Capelatus prykei gen. et sp.n. (Coleoptera: Dytiscidae: Copelatinae) – a phylogenetically isolated diving beetle from the Western Cape of South Africa
Authors: BILTON DAVID T. , TOUSSAINT EMMANUEL F. A. , TURNER CLIVE R. , BALKE MICHAEL
Published: April 2015

Capelatus prykei gen. et sp.n., a distinctive new lineage of copelatine diving beetle, is described from the greater Cape Town area of the Western Cape Province, South Africa, on the basis of both morphological and molecular data. The genus‐level phylogeny of Copelatinae is reconstructed using a combination of nuclear and mitochondrial DNA regions, demonstrating that Capelatus gen.n. has no close relatives within the Afrotropical region, instead forming a clade with the Palaearctic Liopterus and largely Australasian Exocelina. Capelatus gen.n. apparently represents a striking example of a phylogenetically isolated Cape lineage, which also appears to be narrowly endemic and endangered by ongoing habitat loss.
Health economics and outcomes methods in risk‐based decision‐making for blood safety
Authors: Custer Brian , Janssen Mart P. ,   for the Alliance of Blood Operators Risk‐Based Decision‐Making (RBDM) Initiative
Published: April 2015

Analytical methods appropriate for health economic assessments of transfusion safety interventions have not previously been described in ways that facilitate their use. Within the context of risk‐based decision‐making (RBDM), health economics can be important for optimizing decisions among competing interventions. The objective of this review is to address key considerations and limitations of current methods as they apply to blood safety. Because a voluntary blood supply is an example of a public good, analyses should be conducted from the societal perspective when possible. Two primary study designs are recommended for most blood safety intervention assessments: budget impact analysis (BIA), which measures the cost to implement an intervention both to the blood operator but also in a broader context, and cost‐utility analysis (CUA), which measures the ratio between costs and health gain achieved, in terms of reduced morbidity and mortality, by use of an intervention. These analyses often have important limitations because data that reflect specific aspects, for example, blood recipient population characteristics or complication rates, are not available. Sensitivity analyses play an important role. The impact of various uncertain factors can be studied conjointly in probabilistic sensitivity analyses. The use of BIA and CUA together provides a comprehensive assessment of the costs and benefits from implementing (or not) specific interventions. RBDM is multifaceted and impacts a broad spectrum of stakeholders. Gathering and analyzing health economic evidence as part of the RBDM process enhances the quality, completeness, and transparency of decision‐making.
Donation intensity and metabolic syndrome in active whole‐blood donors
Authors: Peffer K. , Verbeek A. L. M. , Swinkels D. W. , Geurts‐Moespot A. J. , den Heijer M. , Atsma F.
Published: April 2015

Background and Objectives

Increased iron and metabolic syndrome (MetS) go hand in hand. Frequent blood donation depletes iron stores. This study investigates whether high‐intensity blood donation is associated with lower MetS prevalence compared with low‐intensity blood donation, and whether iron acts as an intermediary factor.

Materials and Methods

A random sample of 422 male and 211 female active whole‐blood donors ≥45 years of age was included in a cross‐sectional study. Lipids, glucose and iron parameters were measured after overnight fasting. MetS was defined according to the joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention. Three groups of donation intensity were created by sex‐specific tertiles of donation frequency per year and duration of donor career.

Results

MetS was present in 22·9% of donors. Prevalence of MetS was 1·46 (95% confidence interval [CI]: 0·93–2·30) times higher in men with high donation intensity, whereas in women MetS prevalence was 2·14 (95% CI: 0·94–4·86) times higher in donors with high donation intensity compared with those with low donation intensity. In men, increased prevalence of MetS was mainly associated with higher ferritin, whereas high hepcidin predominantly affected MetS prevalence in women.

Conclusion

High‐intensity blood donation is not associated with a decreased prevalence of MetS. In men and women, different iron parameters are associated with MetS prevalence. The temporal relationship between blood donation, iron and MetS, and gender differences herein need to be explored in future research.
Going into surgery: Risk factors for hypertrophic scarring
Authors: Butzelaar Lisolette , Soykan Ezgi A. , Galindo Garre Francisca , Beelen Robert H.J. , Ulrich Magda M. , Niessen Frank B. , Mink van der Molen Aebele B.
Published: April 2015

The aim of this study was to examine the association between possible risk factors for excessive scarring and the formation of hypertrophic scars (HTS). Hypertrophic skin scarring remains a difficult problem in medicine and can cause considerable morbidity. Nevertheless, few extensive prospective studies have been conducted which assess risk factors in relation to HTS formation. Therefore, a prospective observational study was performed. Patients who had elective cardiothoracic surgery were followed for the duration of 1 year and information was collected about a variety of possible risk factors in these patients. The associations between the risk factors and the development of HTSs were investigated. Body mass index, ethnic background, and scar related factors are positively associated with HTS formation. Antihypertensive therapeutics and factors influencing erythropoiesis are negatively associated with HTS formation.
Substrate degradation by the proteasome: A single-molecule kinetic analysis
Authors: Lu Ying , Lee Byung-hoon , King Randall W. , Finley Daniel , Kirschner Marc W.
Published: April 2015

Single-molecule assay of ubiquitylation

Many biological processes in cells are regulated by ubiquitin peptides that are attached to proteins. Measurement of single fluorescent molecules in cell extracts can be used to trace the kinetics of such reactions. Lu
et al.
refined assay conditions to follow ubiquitination by an E3 ubiquitin ligase (see the Perspective by Komander). They visualized the activity of the anaphase-promoting complex (APC), a ubiquitin ligase critical for control of the cell division cycle. The processive initial reaction catalyzed by the APC was replaced by slower reactions. The results show how small, commonly occurring recognition motifs can guide specific and highly controlled enzymatic events. In a companion paper, Lu
et al.
explored how the number and arrangement of added ubiquitin chains affected the interaction of ubiquitylated proteins with the proteasome (a protein complex that recognizes ubiquitylated proteins and degrades them). The extent of ubiquitylation determined the strength of interaction of a substrate protein with the proteasome, and the arrangement of the ubiquitin chains determined the movement of the protein into the proteasome and thus the rate of degradation.

Science
, this issue
10.1126/science.1248737
,
10.1126/science.1250834
; see also p.
183
Acid oceans cited in Earth's worst die-off
Authors: Hand Eric
Published: April 2015

Signature of acidification found in Permian extinctions 250 million years ago.
Thinking inside the box
Authors: Hand Eric
Published: April 2015

A look at the history of the CubeSat.
Ocean acidification and the Permo-Triassic mass extinction
Authors: Clarkson M. O. , Kasemann S. A. , Wood R. A. , Lenton T. M. , Daines S. J. , Richoz S. , Ohnemueller F. , Meixner A. , Poulton S. W. , Tipper E. T.
Published: April 2015

Ocean acidification and mass extinction

The largest mass extinction in Earth's history occurred at the Permian-Triassic boundary 252 million years ago. Several ideas have been proposed for what devastated marine life, but scant direct evidence exists. Clarkson
et al.
measured boron isotopes across this period as a highly sensitive proxy for seawater pH. It appears that, although the oceans buffered the acidifiying effects of carbon release from contemporary pulses of volcanism, buffering failed when volcanism increased during the formation of the Siberian Traps. The result was a widespread drop in ocean pH and the elimination of shell-forming organisms.

Science
, this issue p.
229
Common variants spanning
PLK4
are associated with mitotic-origin aneuploidy in human embryos
Authors: McCoy Rajiv C. , Demko Zachary , Ryan Allison , Banjevic Milena , Hill Matthew , Sigurjonsson Styrmir , Rabinowitz Matthew , Fraser Hunter B. , Petrov Dmitri A.
Published: April 2015

Chromosome number varies in humans

Pregnancy loss is often associated with a loss of chromosome number, a condition known as aneuploidy. When examining aneuploid embryos during in vitro fertilization cycles, McCoy
et al.
found a large genomic region associated with defects in maternal chromosome number (see the Perspective by Vohr and Green). This region contains a gene,
Polo-like Kinase 4
(
PLK4
), that is known to affect chromosome segregation and has variants that correlate with an increased rate of maternal aneuploidy. Surprisingly, such variants occur at relatively high levels in human populations and may be under positive selection.

Science
, this issue p.
235
; see also p.
180
Transatlantic lessons in regulation of mitochondrial replacement therapy
Authors: Cohen I. Glenn , Savulescu Julian , Adashi Eli Y.
Published: April 2015

The UK has approved MRT for clinical use, but the discussion has just begun in the U.S.
Severe Ketoalkalosis as Initial Presentation of Imported Human Rabies in France
Authors: Contou Damien , Dacheux Laurent , Bendib Inès , Jolivet Sarah , Rodriguez Christophe , Tomberli Françoise , Cleret de Langavant Laurent , Lavenir Rachel , Lepelletier Anthony , Larrous Florence , Troupin Cécile , Bourhy Hervé , Brun-Buisson Christian
Published: April 2015

We report a patient with an unusual initial metabolic presentation of imported human rabies who became symptomatic within 2 weeks of returning from Mali to France. This is the single case of imported human rabies identified in France within the past 11 years and the first report of viral RNA in bronchial secretions.
Genetic Determinants of Japanese Encephalitis Virus Vaccine Strain SA14-14-2 That Govern Attenuation of Virulence in Mice
Authors: Gromowski Gregory D. , Firestone Cai-Yen , Whitehead Stephen S.
Published: April 2015

The safety and efficacy of the live-attenuated Japanese encephalitis virus (JEV) SA14-14-2 vaccine are attributed to mutations that accumulated in the viral genome during its derivation. However, little is known about the contribution that is made by most of these mutations to virulence attenuation and vaccine immunogenicity. Here, we generated recombinant JEV (rJEV) strains containing JEV SA14-14-2 vaccine-specific mutations that are located in the untranslated regions (UTRs) and seven protein genes or are introduced from PCR-amplified regions of the JEV SA14-14-2 genome. The resulting mutant viruses were evaluated in tissue culture and in mice. The authentic JEV SA14-14-2 (E) protein, with amino acid substitutions L107F, E138K, I176V, T177A, E244G, Q264H, K279M, A315V, S366A, and K439R relative to the wild-type rJEV clone, was essential and sufficient for complete attenuation of neurovirulence. Individually, the nucleotide substitution T39A in the 5′ UTR (5′-UTR-T39A), the capsid (C) protein amino acid substitution L66S (C-L66S), and the complete NS1/2A genome region containing 10 mutations each significantly reduced virus neuroinvasion but not neurovirulence. The levels of peripheral virulence attenuation imposed by the 5′-UTR-T39A and C-L66S mutations, individually, were somewhat mitigated in combination with other vaccine strain-specific mutations, which might be compensatory, and together did not affect immunogenicity. However, a marked reduction in immunogenicity was observed with the addition of the NS1/2A and NS5 vaccine virus genome regions. These results suggest that a second-generation recombinant vaccine can be rationally engineered to maximize levels of immunogenicity without compromising safety.

IMPORTANCE
The live-attenuated JEV SA14-14-2 vaccine has been vital for controlling the incidence of disease caused by JEV, particularly in rural areas of Asia where it is endemic. The vaccine was developed >25 years ago by passaging wild-type JEV strain SA14 in tissue cultures and rodents, with intermittent tissue culture plaque purifications, to produce a virus clone that had adequate levels of attenuation and immunogenicity. The vaccine and parent virus sequences were later compared, and mutations were identified throughout the vaccine virus genome, but their contributions to attenuation were never fully elucidated. Here, using reverse genetics, we comprehensively defined the impact of JEV SA14-14-2 mutations on attenuation of virulence and immunogenicity in mice. These results are relevant for quality control of new lots of the current live-attenuated vaccine and provide insight for the rational design of second-generation, live-attenuated, recombinant JEV vaccine candidates.
Genetic Confirmation that the H5 Protein Is Required for Vaccinia Virus DNA Replication
Authors: Boyle Kathleen A. , Greseth Matthew D. , Traktman Paula
Published: April 2015

The duplication of the poxvirus double-stranded DNA genome occurs in cytoplasmic membrane-delimited factories. This physical autonomy from the host nucleus suggests that poxvirus genomes encode the full repertoire of proteins committed for genome replication. Biochemical and genetic analyses have confirmed that six viral proteins are required for efficient DNA synthesis; indirect evidence has suggested that the multifunctional H5 protein may also have a role. Here we show that H5 localizes to replication factories, as visualized by immunofluorescence and immunoelectron microscopy, and can be retrieved upon purification of the viral polymerase holoenzyme complex. The temperature-sensitive (
ts
) mutant D
ts
57, which was generated by chemical mutagenesis and has a lesion in H5, exhibits defects in DNA replication and morphogenesis under nonpermissive conditions, depending upon the experimental protocol. The H5 variant encoded by the genome of this mutant is
ts
for function but not stability. For a more precise investigation of how H5 contributes to DNA synthesis, we placed the
ts
57 H5 allele in an otherwise wild-type viral background and also performed small interfering RNA-mediated depletion of H5. Finally, we generated a complementing cell line, CV-1–H5, which allowed us to generate a viral recombinant in which the H5 open reading frame was deleted and replaced with mCherry (vΔH5). Analysis of vΔH5 allowed us to demonstrate conclusively that viral DNA replication is abrogated in the absence of H5. The loss of H5 does not compromise the accumulation of other early viral replication proteins or the uncoating of the virion core, suggesting that H5 plays a direct and essential role in facilitating DNA synthesis.

IMPORTANCE
Variola virus, the causative agent of smallpox, is the most notorious member of the
Poxviridae
family. Poxviruses are unique among DNA viruses that infect mammalian cells, in that their replication is restricted to the cytoplasm of the cell. This physical autonomy from the nucleus has both cell biological and genetic ramifications. Poxviruses must establish cytoplasmic niches that support replication, and the genomes must encode the repertoire of proteins necessary for genome synthesis. Here we focus on H5, a multifunctional and abundant viral protein. We confirm that H5 associates with the DNA polymerase holoenzyme and localizes to the sites of DNA synthesis. By generating an H5-expressing cell line, we were able to isolate a deletion virus that lacks the H5 gene and show definitively that genome synthesis does not occur in the absence of H5. These data support the hypothesis that H5 is a crucial participant in cytoplasmic poxvirus genome replication.
The Greater Affinity of JC Polyomavirus Capsid for α2,6-Linked Lactoseries Tetrasaccharide c than for Other Sialylated Glycans Is a Major Determinant of Infectivity
Authors: Ströh Luisa J. , Maginnis Melissa S. , Blaum Bärbel S. , Nelson Christian D. S. , Neu Ursula , Gee Gretchen V. , O'Hara Bethany A. , Motamedi Nasim , DiMaio Daniel , Atwood Walter J. , Stehle Thilo
Published: April 2015

The human JC polyomavirus (JCPyV) establishes an asymptomatic, persistent infection in the kidneys of the majority of the population and is the causative agent of the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML) in immunosuppressed individuals. The Mad-1 strain of JCPyV, a brain isolate, was shown earlier to require α2,6-linked sialic acid on the lactoseries tetrasaccharide c (LSTc) glycan for attachment to host cells. In contrast, a JCPyV kidney isolate type 3 strain, WT3, has been reported to interact with sialic acid-containing gangliosides, but the role of these glycans in JCPyV infection has remained unclear. To help rationalize these findings and probe the effects of strain-specific differences on receptor binding, we performed a comprehensive analysis of the glycan receptor specificities of these two representative JCPyV strains using high-resolution X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy, and correlated these data with the results of infectivity assays. We show here that capsid proteins of Mad-1 and WT3 JCPyV can both engage LSTc as well as multiple sialylated gangliosides. However, the binding affinities exhibit subtle differences, with the highest affinity observed for LSTc. Engagement of LSTc is a prerequisite for functional receptor engagement, while the more weakly binding gangliosides are not required for productive infection. Our findings highlight the complexity of virus-carbohydrate interactions and demonstrate that subtle differences in binding affinities, rather than the binding event alone, help determine tissue tropism and viral pathogenesis.

IMPORTANCE
Viral infection is initiated by attachment to receptors on host cells, and this event plays an important role in viral disease. We investigated the receptor-binding properties of human JC polyomavirus (JCPyV), a virus that resides in the kidneys of the majority of the population and can cause the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML) in the brains of immunosuppressed individuals. JCPyV has been reported to interact with multiple carbohydrate receptors, and we sought to clarify how the interactions between JCPyV and cellular carbohydrate receptors influenced infection. Here we demonstrate that JCPyV can engage numerous sialylated carbohydrate receptors. However, the virus displays preferential binding to LSTc, and only LSTc mediates a productive infection. Our findings demonstrate that subtle differences in binding affinity, rather than receptor engagement alone, are a key determinant of viral infection.
Intercontinental Spread of Asian-Origin H5N8 to North America through Beringia by Migratory Birds
Authors: Lee Dong-Hun , Torchetti Mia Kim , Winker Kevin , Ip Hon S. , Song Chang-Seon , Swayne David E.
Published: April 2015

Phylogenetic network analysis and understanding of waterfowl migration patterns suggest that the Eurasian H5N8 clade 2.3.4.4 avian influenza virus emerged in late 2013 in China, spread in early 2014 to South Korea and Japan, and reached Siberia and Beringia by summer 2014 via migratory birds. Three genetically distinct subgroups emerged and subsequently spread along different flyways during fall 2014 into Europe, North America, and East Asia, respectively. All three subgroups reappeared in Japan, a wintering site for waterfowl from Eurasia and parts of North America.
HIV Blocks Interferon Induction in Human Dendritic Cells and Macrophages by Dysregulation of TBK1
Authors: Harman Andrew N. , Nasr Najla , Feetham Alexandra , Galoyan Ani , Alshehri Abdullateef A. , Rambukwelle Dharshini , Botting Rachel A. , Hiener Bonnie M. , Diefenbach Eve , Diefenbach Russell J. , Kim Min , Mansell Ashley , Cunningham Anthony L.
Published: April 2015

Dendritic cells (DCs) and macrophages are present in the tissues of the anogenital tract, where HIV-1 transmission occurs in almost all cases. These cells are both target cells for HIV-1 and represent the first opportunity for the virus to interfere with innate recognition. Previously we have shown that both cell types fail to produce type I interferons (IFNs) in response to HIV-1 but that, unlike T cells, the virus does not block IFN induction by targeting IFN regulatory factor 3 (IRF3) for cellular degradation. Thus, either HIV-1 inhibits IFN induction by an alternate mechanism or, less likely, these cells fail to sense HIV-1. Here we show that HIV-1 (but not herpes simplex virus 2 [HSV-2] or Sendai virus)-exposed DCs and macrophages fail to induce the expression of all known type I and III IFN genes. These cells do sense the virus, and pattern recognition receptor (PRR)-induced signaling pathways are triggered. The precise stage in the IFN-inducing signaling pathway that HIV-1 targets to block IFN induction was identified; phosphorylation but not K63 polyubiquitination of TANK-binding kinase 1 (TBK1) was completely inhibited. Two HIV-1 accessory proteins, Vpr and Vif, were shown to bind to TBK1, and their individual deletion partly restored IFN-β expression. Thus, the inhibition of TBK1 autophosphorylation by binding of these proteins appears to be the principal mechanism by which HIV-1 blocks type I and III IFN induction in myeloid cells.

IMPORTANCE
Dendritic cells (DCs) and macrophages are key HIV target cells. Therefore, definition of how HIV impairs innate immune responses to initially establish infection is essential to design preventative interventions, especially by restoring initial interferon production. Here we demonstrate how HIV-1 blocks interferon induction by inhibiting the function of a key kinase in the interferon signaling pathway, TBK1, via two different viral accessory proteins. Other viral proteins have been shown to target the general effects of TBK1, but this precise targeting between ubiquitination and phosphorylation of TBK1 is novel.
A Single Amino Acid Deletion in the Matrix Protein of Porcine Reproductive and Respiratory Syndrome Virus Confers Resistance to a Polyclonal Swine Antibody with Broadly Neutralizing Activity
Authors: Trible Benjamin R. , Popescu Luca N. , Monday Nicholas , Calvert Jay G. , Rowland Raymond R. R.
Published: April 2015

Assessment of virus neutralization (VN) activity in 176 pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) identified one pig with broadly neutralizing activity. A Tyr-10 deletion in the matrix protein provided escape from broad neutralization without affecting homologous neutralizing activity. The role of the Tyr-10 deletion was confirmed through an infectious clone with a Tyr-10 deletion. The results demonstrate differences in the properties and specificities of VN responses elicited during PRRSV infection.
Transformation with Oncogenic Ras and the Simian Virus 40 T Antigens Induces Caspase-Dependent Sensitivity to Fatty Acid Biosynthetic Inhibition
Authors: Xu Shihao , Spencer Cody M. , Munger Joshua
Published: April 2015

Oncogenesis is frequently accompanied by the activation of specific metabolic pathways. One such pathway is fatty acid biosynthesis, whose induction is observed upon transformation of a wide variety of cell types. Here, we explored how defined oncogenic alleles, specifically the simian virus 40 (SV40) T antigens and oncogenic Ras
12V
, affect fatty acid metabolism. Our results indicate that SV40/Ras
12V
-mediated transformation of fibroblasts induces fatty acid biosynthesis in the absence of significant changes in the concentration of fatty acid biosynthetic enzymes. This oncogene-induced activation of fatty acid biosynthesis was found to be mammalian target of rapamycin (mTOR) dependent, as it was attenuated by rapamycin treatment. Furthermore, SV40/Ras
12V
-mediated transformation induced sensitivity to treatment with fatty acid biosynthetic inhibitors. Pharmaceutical inhibition of acetyl-coenzyme A (CoA) carboxylase (ACC), a key fatty acid biosynthetic enzyme, induced caspase-dependent cell death in oncogene-transduced cells. In contrast, isogenic nontransformed cells were resistant to fatty acid biosynthetic inhibition. This oncogene-induced sensitivity to fatty acid biosynthetic inhibition was independent of the cells' growth rates and could be attenuated by supplementing the medium with unsaturated fatty acids. Both the activation of fatty acid biosynthesis and the sensitivity to fatty acid biosynthetic inhibition could be conveyed to nontransformed breast epithelial cells through transduction with oncogenic Ras
12V
. Similar to what was observed in the transformed fibroblasts, the Ras
12V
-induced sensitivity to fatty acid biosynthetic inhibition was independent of the proliferative status and could be attenuated by supplementing the medium with unsaturated fatty acids. Combined, our results indicate that specific oncogenic alleles can directly confer sensitivity to inhibitors of fatty acid biosynthesis.

IMPORTANCE
Viral oncoproteins and cellular mutations drive the transformation of normal cells to the cancerous state. These oncogenic alterations induce metabolic changes and dependencies that can be targeted to kill cancerous cells. Here, we find that the cellular transformation resulting from combined expression of the SV40 early region with an oncogenic Ras allele is sufficient to induce cellular susceptibility to fatty acid biosynthetic inhibition. Inhibition of fatty acid biosynthesis in these cells resulted in programmed cell death, which could be rescued by supplementing the medium with nonsaturated fatty acids. Similar results were observed with the expression of oncogenic Ras in nontransformed breast epithelial cells. Combined, our results suggest that specific oncogenic alleles induce metabolic dependencies that can be exploited to selectively kill cancerous cells.
Systematic review: unmet supportive care needs in people diagnosed with chronic liver disease
Authors: Valery Patricia C , Powell Elizabeth , Moses Neta , Volk Michael L , McPhail Steven M , Clark Paul J , Martin Jennifer
Published: April 2015

Objective

People with chronic liver disease, particularly those with decompensated cirrhosis, experience several potentially debilitating complications that can have a significant impact on activities of daily living and quality of life. These impairments combined with the associated complex treatment mean that they are faced with specific and high levels of supportive care needs. We aimed to review reported perspectives, experiences and concerns of people with chronic liver disease worldwide. This information is necessary to guide development of policies around supportive needs screening tools and to enable prioritisation of support services for these patients.

Design

Systematic searches of PubMed, MEDLINE, CINAHL and PsycINFO from the earliest records until 19 September 2014. Data were extracted using standardised forms. A qualitative, descriptive approach was utilised to analyse and synthesise data.

Results

The initial search yielded 2598 reports: 26 studies reporting supportive care needs among patients with chronic liver disease were included, but few of them werepatient-reportedneeds, none used a validated liver disease-specific supportive care need assessment instrument, and only three included patients with cirrhosis. Five key domains of supportive care needs were identified: informational or educational (eg, educational material, educational sessions), practical (eg, daily living), physical (eg, controlling pruritus and fatigue), patient care and support (eg, support groups), and psychological (eg, anxiety, sadness).

Conclusions

While several key domains of supportive care needs were identified, most studies included hepatitis patients. There is a paucity of literature describing the supportive care needs of the chronic liver disease population likely to have the most needs—namely those with cirrhosis. Assessing the supportive care needs of people with chronic liver disease have potential utility in clinical practice for facilitating timely referrals to support services.
Secondhand smoke in outdoor settings: smokers’ consumption, non-smokers’ perceptions, and attitudes towards smoke-free legislation in Spain
Authors: Sureda Xisca , Fernández Esteve , Martínez-Sánchez Jose M , Fu Marcela , López María J , Martínez Cristina , Saltó Esteve
Published: April 2015

Objective

To describe where smokers smoke outdoors, where non-smokers are exposed outdoors to secondhand smoke (SHS), and attitudes towards smoke-free outdoor areas after the implementation of national smoke-free legislation.

Design

This cross-sectional study was conducted between June 2011 and March 2012 (n=1307 participants).

Setting

Barcelona, Spain.

Participants

Representative, random sample of the adult (≥16 years) population.

Primary and secondary outcomes

Proportion of smoking and prevalence of exposure to SHS in the various settings according to type of enclosure. Percentages of support for outdoor smoke-free policies according to smoking status.

Results

Smokers reported smoking outdoors most in bars and restaurants (54.8%), followed by outdoor places at work (46.8%). According to non-smokers, outdoor SHS exposure was highest at home (42.5%) and in bars and restaurants (33.5%). Among non-smoking adult students, 90% claimed exposure to SHS on university campuses. There was great support for banning smoking in the majority of outdoor areas, which was stronger among non-smokers than smokers. Over 70% of participants supported smoke-free playgrounds, school and high school courtyards, and the grounds of healthcare centres.

Conclusions

Extending smoking bans to selected outdoor settings should be considered in further tobacco control interventions to protect non-smokers from SHS exposure and to establish a positive model for youth. The majority of public support for some outdoor smoke-free areas suggests that it is feasible to extend smoking bans to additional outdoor settings.
Richard's back: death, scoliosis and myth making
Authors: Lund Mary Ann
Published: April 2015

The body of a mediaeval monarch was always under scrutiny, and Richard III's was no exception. In death, however, his body became subject to new forms of examination and interpretation: stripped naked after the battle of Bosworth, his corpse was carried to Leicester and exhibited before being buried. In 2012, it was rediscovered. The revelation that Richard suffered from scoliosis prompts this article to re-evaluate the historical sources about Richard's physique and his posthumous reputation. This article argues that Richard's death and his myth as ‘crookback’ are inextricably linked and traces attitudes to spinal curvature in the early modern period. It also considers how Shakespeare represented Richard as deformed, and aspects of performance history which suggest physical vulnerability. It then considers Richard's scoliosis from the perspective of medical history, reviewing classical accounts of scoliosis and arguing that Richard was probably treated with a mixture of axial traction and pressure. It demonstrates from the evidence of Richard's medical household that he was well placed to receive hands-on therapies and considers in particular the role of his physician and surgeon, William Hobbes. Finally, it shows how the case of Richard III demonstrates the close relationship between politics and medicine in the period and the contorted process of historical myth making.
Pipeline embolization for definitive endoluminal reconstruction of blister-type carotid aneurysms after clip wrapping
Authors: Kalani M Yashar S , Albuquerque Felipe C , Levitt Michael , Nakaji Peter , Spetzler Robert F , McDougall Cameron
Published: April 2015

Background

Ruptured blister aneurysms of the carotid artery are difficult to safely treat. We present a novel strategy of microsurgical clip wrapping of internal carotid artery blister aneurysms in the setting of acute rupture, followed by delayed placement of a pipeline embolization device for definitive treatment.

Clinical presentation

We present two cases of ruptured blister aneurysms of the internal carotid artery treated by wrapping of the diseased segment of the vessel, followed by delayed deployment of a flow diverting stent once the patient was out of the vasospasm window but during the same hospitalization.

Results

Clip wrapping followed by flow diversion in a delayed fashion results in anatomic remodeling of the diseased artery without a high morbidity.

Conclusions

A combined approach of acute surgical stabilization followed by definitive endovascular reconstruction may reduce hemorrhagic complications while improving long term treatment durability.