bluuebunny/crossref_metadata_2025 huggingface dataset:
Biorevitalizing effect of a novel facial serum containing apple stem cell extract, pro‐collagen lipopeptide, creatine, and urea on skin aging signs
Authors: Sanz Maria Teresa , Campos Celia , Milani Massimo , Foyaca Monica , Lamy Amandine , Kurdian Karine , Trullas Carles
Published: October 2015

Background

Epithelial regeneration in skin is achieved by the constant turnover and differentiation of keratinocytes. Epidermal and dermal stem cells compartments are fundamental for the continuous renewal of the skin. Adult stem cells are the unique source for skin tissue renewal. Plants have stem cells and plant derived stem cell extracts are now used in topical products for their potential anti‐ageing and anti‐wrinkle effects. A new dermocosmetic product containing apple stem cell extract, urea, creatine and palmitoyl tripeptide‐38 (Ureadin Fusion Serum Lift Antiarrugas, ISDIN S.A), has been recently developed to target different aspects involved in skin aging.

Study aim

To assess in vitro the effects of this new serum on the metabolic functions of human senescent fibroblasts and in vivo the anti‐aging effects by clinical and instrumental evaluation.

Methods

We evaluated the effects of the serum on the mitochondrial ROS (reactive oxygen species) production in human senescent cultured fibroblasts measured at 0.1% and 1% using the Mitoread AntiOx mtROS method. In addition we evaluated the anti‐ageing in vivo effect of this new serum applied on the face twice daily for 28 consecutive days and assessed by clinical and instrumental evaluation in 32 women with sensitive skin bearing wrinkles on crow's feet.

Results

The tested serum both at 0.1% and 1% induces a significant increase in 02 consumption, cellular ATP level and a reduction in extra‐cellular lactate concentration. The product reduces also significantly the mitochondrial ROS production. The clinical study shows a relevant anti‐wrinkle effect in 71% of the treated women with visible effects in 68% of the subjects as soon as 7 days of treatment. A significant increase in dermal density and skin elasticity was also observed.

Conclusion

The use of this novel anti‐aging serum demonstrated a significant improvement of aging skin signs with first visible results achieved after one week of use. The product seemed to optimize the metabolic functions in human senescent cultured fibroblast restoring a more efficient cell metabolism therefore contributing to the anti‐aging properties of the product.
The Effect of Luting Cement and Titanium Base on the Final Color of Zirconium Oxide Core Material
Authors: Capa Nuray , Tuncel Ilkin , Tak Onjen , Usumez Aslihan
Published: October 2015

Purpose

To evaluate the effects of different types of luting cements and different colors of zirconium cores on the final color of the restoration that simulates implant‐supported fixed partial dentures (FPDs) by using a titanium base on the bottom.

Materials and Methods

One hundred and twenty zirconium oxide core plates (Zr‐Zahn; 10 mm in width, 5 mm in length, 0.5 mm in height) were prepared in different shades (n = 20; noncolored, A2, A3, B1, C2, D2). The specimens were subdivided into two subgroups for the two types of luting cements (n = 10). The initial color measurements were made on zirconium oxide core plates using a spectrometer. To create the cement thicknesses, stretch strips with holes in the middle (5 mm in diameter, 70 μm in height) were used. The second measurement was done on the zirconium oxide core plates after the application of the resin cement (U‐200, A2 Shade) or polycarboxylate cement (Lumicon). The final measurement was done after placing the titanium discs (5 mm in diameter, 3 mm in height) in the bottom. The data were analyzed with two‐way ANOVA and Tukey's honestly significant differences (HSD) tests (α = 0.05).

Results

The ∆E*ab value was higher in the resin cement‐applied group than in the polycarboxylate cement‐applied group (p < 0.001). The highest ∆E*ab value was recorded for the zirconium oxide core‐resin cement–titanium base, and the lowest was recorded for the polycarboxylate cement–zirconium oxide core (p < 0.001).

Conclusion

The luting cement, the presence of titanium, and the color of zirconium are all important factors that determine the final shade of zirconia cores in implant‐supported FPDs.
Hybrid ‘superswarm’ leads to rapid divergence and establishment of populations during a biological invasion
Authors: Roy Denis , Lucek Kay , Walter Ryan P. , Seehausen Ole
Published: October 2015

Understanding the genetic background of invading species can be crucial information clarifying why they become invasive. Intraspecific genetic admixture among lineages separated in the native ranges may promote the rate and extent of an invasion by substantially increasing standing genetic variation. Here, we examined the genetic relationships among threespine stickleback that recently colonized Switzerland. This invasion results from several distinct genetic lineages that colonized multiple locations and have since undergone range expansions, where they coexist and admix in parts of their range. Using 17 microsatellites genotyped for 634 individuals collected from 17 Swiss and two non‐Swiss European sites, we reconstruct the invasion of stickleback and investigate the potential and extent of admixture and hybridization among the colonizing lineages from a population genetic perspective. Specifically, we test for an increase in standing genetic variation in populations where multiple lineages coexist. We find strong evidence of massive hybridization early on, followed by what appears to be recent increased genetic isolation and the formation of several new genetically distinguishable populations, consistent with a hybrid ‘superswarm’. This massive hybridization and population formation event(s) occurred over approximately 140 years and likely fuelled the successful invasion of a diverse range of habitats. The implications are that multiple colonizations coupled with hybridization can lead to the formation of new stable genetic populations potentially kick‐starting speciation and adaptive radiation over a very short timescale.
Parasite molecules and host responses in cystic echinococcosis
Authors: Díaz A. , Casaravilla C. , Barrios A. A. , Ferreira A. M.
Published: October 2015

Cystic echinococcosis is the infection by the larvae of cestode parasites belonging to the Echinococcus granulosus sensu lato species complex. Local host responses are strikingly subdued in relation to the size and persistence of these larvae, which develop within mammalian organs as ‘hydatid cysts’ measuring up to tens of cm in diameter. In a context in which helminth‐derived immune‐suppressive, as well as Th2‐inducing, molecules garner much interest, knowledge on the interactions between E. granulosus molecules and the immune system lags behind. Here, we discuss what is known and what are the open questions on E. granulosus molecules and structures interacting with the innate and adaptive immune systems, potentially or in demonstrated form. We attempt a global biological approach on molecules that have been given consideration primarily as protective (Eg95) or diagnostic antigens (antigen B, antigen 5). We integrate glycobiological information, which traverses the discussions on antigen 5, the mucin‐based protective laminated layer and immunologically active preparations from protoscoleces. We also highlight some less well‐known molecules that appear as promising candidates to possess immune‐regulatory activities. Finally, we point out gaps in the molecular‐level knowledge of this infectious agent that hinder our understanding of its immunology.
Bridging the gap between interviewer and interviewee: developing an interview guide for individual interviews by means of a focus group
Authors: Pedersen Birgith , Delmar Charlotte , Falkmer Ursula , Grønkjær Mette
Published: October 2015

Background

In developing an interview guide, pre‐existing knowledge about the research topic is essential. In a recent study, we were interested in exploring the experiences of weight changes among women treated for breast cancer using individual interviews. However, to develop an interview guide for the individual interviews that covered relevant thematic and dynamic dimensions, we found existing literature insufficient. Thus, we turned our attention to the benefit of the focus group method.

Objectives

This study aims to discuss how a focus group prior to individual interviews may contribute in developing the thematic dimension and translating the dynamic dimension of an interview guide into everyday language.

Methods

We conducted one focus group interview of five women treated for breast cancer with experiences in weight changes. Data were analysed using content and conversation analysis and discussed with relevant literature on interview guide development.

Ethics

The study is approved by the Danish Data Protection Agency (2008‐58‐0028) and follows the ethical guidelines for qualitative research.

Results

Data generation and analysis resulted in themes for the thematic dimension, as well as three dynamic areas to consider in the individual interviews to bridge the gap between the interviewer and the interviewee. The dynamic areas are as follows: The use of words, images and metaphors – a shield and self‐protection, Multiple meanings to explore and Staying close to the everyday language.

Conclusion

The analysis made us more familiar with the content and meaning of weight changes among breast cancer survivors. Furthermore, it provided images and metaphors, multiple meanings and a sense of the women's everyday language that calls for an open interview frame to be used in subsequent individual interviews.
Beginnings—The Kidney and Nephrology in Ancient Mesopotamian Culture
Authors: Eknoyan Garabed
Published: October 2015

A defining period in the history of civilization occurred in ancient Mesopotamia. While some of Mesopotamian contributions to knowledge (writing, mathematics, astronomy) have been recognized, those made to medicine are just beginning to be studied and appreciated. The medicine of the time developed in a theocratic society where local gods controlled all aspects of life and their healers were learned scribes who established the priestly medicine of old. For their use of prayers and incantations as a component of therapy Mesopotamian medicine has been belittled and relegated to magic. In fact, the Mesopotamian healers established the basic medical skills of observation, diagnosis, prognosis and treatment and, over time, systematically produced an expanding corpus of medical knowledge that had not existed theretofore. The kidney as an organ, the components of the urogenital tract, some signs and symptoms of renal diseases, and the art of urinalysis were first conceptualized, studied, and codified in Mesopotamia. In doing so, the Mesopotamians set the foundations of rational medicine that would follow after the introduction of the phonetic alphabet and the Socratic method of questioning and debate to stimulate critical analysis.
Mortality in adult immigrants in the 2000s in Belgium: a test of the ‘healthy‐migrant’ and the ‘migration‐as‐rapid‐health‐transition’ hypotheses
Authors: Vandenheede Hadewijch , Willaert Didier , De Grande Hannelore , Simoens Steven , Vanroelen Christophe
Published: October 2015

Objective

Firstly, to map out and compare all‐cause and cause‐specific mortality patterns by migrant background in Belgium; and secondly, to probe into explanations for the observed patterns, more specifically into the healthy‐migrant, acculturation and the migration‐as‐rapid‐health‐transition theories.

Methods

Data comprise individually linked Belgian census‐mortality follow‐up data for the period 2001–2011. All official inhabitants aged 25–54 at time of the census were included. To delve into the different explanations, differences in all‐cause and chronic‐ and infectious‐disease mortality were estimated using Poisson regression models, adjusted for age, socioeconomic position and urbanicity.

Results

First‐generation immigrants have lower all‐cause and chronic‐disease mortality than the host population. This mortality advantage wears off with length of stay and is more marked among non‐Western than Western first‐generation immigrants. For example, Western and non‐Western male immigrants residing 10 years or more in Belgium have a mortality rate ratio for cardiovascular disease of 0.72 (95% CI 0.66–0.78) and 0.59 (95% CI 0.53–0.66), respectively (vs host population). The pattern of infectious‐disease mortality in migrants is slightly different, with rather high mortality rates in first‐generation sub‐Saharan Africans and rather low rates in all other immigrant groups. As for second‐generation immigrants, the picture is gloomier, with a mortality disadvantage that disappears after control for socioeconomic position.

Conclusion

Findings are largely consistent with the healthy‐migrant, acculturation and the migration‐as‐rapid‐health‐transition theories. The convergence of the mortality profile of second‐generation immigrants towards that of the host population with similar socioeconomic position indicates the need for policies simultaneously addressing different areas of deprivation.
Potassium inhibits nitric oxide and adenosine arteriolar vasodilatation via KIR and Na+/K+ATPase: implications for redundancy in active hyperaemia
Authors: Lamb Iain R. , Murrant Coral L.
Published: October 2015

Key points

Multiple vasodilators have been identified as being important in matching blood flow to the metabolism of contracting skeletal muscle and it has been hypothesized that this process is governed by redundancy between vasodilators, where one vasodilator can compensate for the loss of another.

In the present study, we aimed to determine whether redundancy between vasodilators exists by investigating whether vasodilators relevant to skeletal muscle contraction can inhibit the effects of other vasodilators.

We show that potassium can inhibit vasodilatations induced by adenosine and nitric oxide, and also that adenosine and nitric oxide can interact in a way that changes over time.

Furthermore, we show that inward rectifying potassium channels and Na+/K+ATPase are partially mechanistically responsible for the interaction between potassium and adenosine and nitric oxide.

Our data provide proof of principle that vasodilators relevant to muscle contraction interact and also that redundancy may govern the processes of active hyperaemia.

Redundancy, in active hyperaemia, where one vasodilator can compensate for another if the first is missing, would require that one vasodilator inhibits the effects of another; therefore, if the first vasodilator is inhibited, its inhibitory influence on the second vasodilator is removed and the second vasodilator exerts a greater vasodilatory effect. We aimed to determine whether vasodilators relevant to skeletal muscle contraction [potassium chloride (KCl), adenosine (ADO) and nitric oxide] inhibit one another and, in addition, to investigate the mechanisms for this interaction. We used the hamster cremaster muscle and intravital microscopy to directly visualize 2A arterioles when exposed to a range of concentrations of one vasodilator [10−8 to 10−5 m S‐nitroso‐N‐acetyl penicillamine (SNAP), 10−8 to 10−5 m ADO, 10 and 20 mm KCl] in the absence and then in the presence of a second vasodilator (10−7 m ADO, 10−7 m SNAP, 10 mm KCl). We found that KCl significantly attenuated SNAP‐induced vasodilatations by ∼65.8% and vasodilatations induced by 10−8 to 10−6 m ADO by ∼72.8%. Furthermore, we observed that inhibition of KCl vasodilatation, by antagonizing either Na+/K+ ATPase using ouabain or inward rectifying potassium channels using barium chloride, could restore the SNAP‐induced vasodilatation by up to ∼53.9% and 30.6%, respectively, and also restore the ADO‐induced vasodilatations by up to ∼107% and 76.7%, respectively. Our data show that vasodilators relevant to muscle contraction can interact in a way that alters the effectiveness of other vasodilators. These data suggest that active hyperaemia may be the result of complex interactions between multiple vasodilators via a redundant control paradigm.
Effects of conducting defects on resistive switching characteristics of SiNx-based resistive random-access memory with MIS structure
Authors: Kim Sungjun , Cho Seongjae , Ryoo Kyung-Chang , Park Byung-Gook
Published: October 2015

In this work, the effects of conducting defects on resistive switching characteristics of SiNx-based resistive random-access memory (RRAM) have been investigated. Two types of RRAM devices having metal–insulator–silicon layer configuration were fabricated. One is the device with SiNx as the resistive switching layer deposited by plasma-enhanced chemical vapor deposition (PECVD), and the other has the SiNx layer prepared by low-pressure chemical vapor deposition (LPCVD). The device cell deposited by LPCVD (LP-SiNx cell afterward) demonstrated superior uniformity of switching parameters and better endurance cycles compared with the device cell deposited by PECVD (PE-SiNx cell afterward).
High temperature coefficient of resistance achieved by ion beam assisted sputtering with no heat treatment in VyM1−yOx (M = Nb, Hf)
Authors: Vardi Naor , Sharoni Amos
Published: October 2015

Thermal imaging based on room temperature bolometer sensors is a growing market, constantly searching for improved sensitivity. One important factor is the temperature coefficient of resistance (TCR), i.e., the sensitivity of the active material. Herein, the authors report the improved TCR properties attainable by the “ion beam assisted deposition” method for room temperature deposition. VyM1−yOx (M = Nb, Hf) thin-film alloys were fabricated on 1 μm thermal SiO2 atop Si (100) substrates by reactive magnetron cosputtering at room temperature using a low energy ion source, aimed at the film, to insert dissociated oxygen species and increase film density. The authors studied the influence of deposition parameters such as oxygen partial pressure, V to M ratio, and power of the plasma source, on resistance and TCR. The authors show high TCR (up to −3.7% K−1) at 300 K, and excellent uniformity, but also an increase in resistance. The authors emphasize that samples were prepared at room temperature with no heat treatment, much simpler than common processes that require annealing at high temperatures. So, this is a promising fabrication route for uncooled microbolometers.
3D localization of electrophysiology catheters from a single x‐ray cone‐beam projection
Authors: Robert Normand , Polack George G. , Sethi Benu , Rowlands John A. , Crystal Eugene
Published: October 2015

Purpose:

X‐ray images allow the visualization of percutaneous devices such as catheters in real time but inherently lack depth information. The provision of 3D localization of these devices from cone beam x‐ray projections would be advantageous for interventions such as electrophysiology (EP), whereby the operator needs to return a device to the same anatomical locations during the procedure. A method to achieve real‐time 3D single view localization (SVL) of an object of known geometry from a single x‐ray image is presented. svl exploits the change in the magnification of an object as its distance from the x‐ray source is varied. The x‐ray projection of an object of interest is compared to a synthetic x‐ray projection of a model of said object as its pose is varied.

Methods:

svl was tested with a 3 mm spherical marker and an electrophysiology catheter. The effect of x‐ray acquisition parameters on svl was investigated. An independent reference localization method was developed to compare results when imaging a catheter translated via a computer controlled three‐axes stage. svl was also performed on clinical fluoroscopy image sequences. A commercial navigation system was used in some clinical image sequences for comparison.

Results:

svl estimates exhibited little change as x‐ray acquisition parameters were varied. The reproducibility of catheter position estimates in phantoms denoted by the standard deviations, (σx, σy, σz) = (0.099 mm,  0.093 mm,  2.2 mm), where x and y are parallel to the detector plane and z is the distance from the x‐ray source. Position estimates (x, y, z) exhibited a 4% systematic error (underestimation) when compared to the reference method. The authors demonstrated that EP catheters can be tracked in clinical fluoroscopic images.

Conclusions:

It has been shown that EP catheters can be localized in real time in phantoms and clinical images at fluoroscopic exposure rates. Further work is required to characterize performance in clinical images as well as the sensitivity to clinical image quality.
New proteins may expand, improve genome editing
Authors: Pennisi Elizabeth
Published: October 2015

Mining bacterial immune systems yields two new editing tools and has the potential to contribute many more.
Ultrafast 2D IR spectroscopy of the excess proton in liquid water
Authors: Thämer Martin , De Marco Luigi , Ramasesha Krupa , Mandal Aritra , Tokmakoff Andrei
Published: October 2015

How well does water share its protons?

Chemists have spent centuries trying to understand what acids look like at the molecular level. It's clear now that water molecules in the liquid accommodate extra protons. Less clear is whether the protons piggyback on individual water molecules (Eigen structure) or find shared accommodation between two at a time (Zundel structure). Thämer
et al.
acquired time-resolved vibrational spectra across an unusually broad span of the mid-infrared, allowing them to monitor stretches and bends at the same time. Their results imply a more prominent role for the Zundel structure than previously anticipated.

Science
, this issue p.
78
The IPCC at a crossroads: Opportunities for reform
Authors: Carraro Carlo , Edenhofer Ottmar , Flachsland Christian , Kolstad Charles , Stavins Robert , Stowe Robert
Published: October 2015

Increase focus on policy-relevant research
Genetic determinants of in vivo fitness and diet responsiveness in multiple human gut
Bacteroides
Authors: Wu Meng , McNulty Nathan P. , Rodionov Dmitry A. , Khoroshkin Matvei S. , Griffin Nicholas W. , Cheng Jiye , Latreille Phil , Kerstetter Randall A. , Terrapon Nicolas , Henrissat Bernard , Osterman Andrei L. , Gordon Jeffrey I.
Published: October 2015

Diet shapes host and gut microbe fitness

The human gut microbiota is hugely diverse, with many strain variants having a multiplicity of effects on host metabolism and immunity. To define some of these functions, Wu
et al.
made libraries of mutants of Bacteroides species known for their capacity to process otherwise intractable dietary fiber. Germ-free mice colonized with defined gut microbiota communities containing the mutants were fed specific diets containing different ratios of fat and fiber. Genes, strains, and species were identified that were associated with specific metabolic pathways. The community responses to dietary shifts were manipulated in an attempt to characterize species for their probiotic or therapeutic potential.

Science
, this issue
10.1126/science.aac5992
>
End-bonded contacts for carbon nanotube transistors with low, size-independent resistance
Authors: Cao Qing , Han Shu-Jen , Tersoff Jerry , Franklin Aaron D. , Zhu Yu , Zhang Zhen , Tulevski George S. , Tang Jianshi , Haensch Wilfried
Published: October 2015

Making better small contacts

Semiconducting single-walled carbon nanotubes have potential size and conductivity advantages over silicon for making smaller transistors. However, as metal electrical contacts decrease in size, the associated resistance increases to impractical values. Cao
et al.
reacted molybdenum films with semiconducting carbon nanotubes to create a carbide contact. The resistance of these contacts remained low even for 10-nm-scale contacts.

Science
, this issue p.
68
It takes the world to understand the brain
Authors: Huang Z. Josh , Luo Liqun
Published: October 2015

International brain projects discuss how to coordinate efforts
Comparative Evaluation of Three Nucleic Acid-Based Assays for BK Virus Quantification
Authors: Descamps Veronique , Martin Elodie , Morel Virginie , François Catherine , Helle François , Duverlie Gilles , Castelain Sandrine , Brochot Etienne
Published: October 2015

With the growing importance of BK virus (BKV), effective and efficient screening for BKV replication in plasma and urine samples is very important for monitoring renal transplant and hematopoietic stem cell transplant recipients, who are at increased risk of BKV-associated diseases. However, recent assays proposed by many manufacturers have not been tested, and the available tests have not been standardized. The aim of the present study was to evaluate and compare the performances of three commercially available kits, R-gene, GeneProof, and RealStar, on plasma and urine specimens from patients infected with various genotypes and to determine the correlations with the results from a reference laboratory. A qualitatively excellent global agreement (96.8%) was obtained. RealStar PCR tended to give a higher sensitivity, especially for subtype Ib1 samples. Comparison of 30 plasma samples and 53 urine samples showed a good agreement between the three assays, with Spearman's Rho correlation coefficient values falling between 0.92 and 0.98 (
P
< 0.001). Moreover, a perfect correlation was obtained for comparison of the assay performances with the AcroMetrix BKV panel (
P
< 0.001 for all comparisons). According to Bland-Altman analysis, more than 95% (240/249 comparisons) of sample comparisons were situated in the range of the mean ± 2 standard deviations (SD). The greatest variability between assays was observed for 10.2% of subtype Ib2 samples, with differences of >1 log
10
copies/ml. In conclusion, this study demonstrated the reliable and comparable performances of the R-gene, GeneProof, and RealStar real-time PCR systems for quantification of BKV in urine and plasma samples. All three real-time PCR assays are appropriate for screening of BKV replication in patients.
Characterization of Five Zoonotic Streptococcus suis Strains from Germany, Including One Isolate from a Recent Fatal Case of Streptococcal Toxic Shock-Like Syndrome in a Hunter
Authors: Eisenberg Tobias , Hudemann Christoph , Hossain Hamid M. , Hewer Angela , Tello Khodr , Bandorski Dirk , Rohde Manfred , Valentin-Weigand Peter , Baums Christoph Georg
Published: October 2015

A
Streptococcus suis
isolate from a German hunter with streptococcal toxic shock-like syndrome (STSLS) and four additional zoonotic isolates were genotyped as
mrp
+
epf
* (variant 1890)
sly
+
cps2
+
. All five zoonotic German strains were characterized by high multiplication in human blood samples
ex vivo
, but induction of only low levels of proinflammatory cytokines compared to a Chinese STSLS strain.
MDA5 Is Critical to Host Defense during Infection with Murine Coronavirus
Authors: Zalinger Zachary B. , Elliott Ruth , Rose Kristine M. , Weiss Susan R.
Published: October 2015

Infection with the murine coronavirus mouse hepatitis virus (MHV) activates the pattern recognition receptors melanoma differentiation-associated gene 5 (MDA5) and Toll-like receptor 7 (TLR7) to induce transcription of type I interferon. Type I interferon is crucial for control of viral replication and spread in the natural host, but the specific contributions of MDA5 signaling to this pathway as well as to pathogenesis and subsequent immune responses are largely unknown. In this study, we use MHV infection of the liver as a model to demonstrate that MDA5 signaling is critically important for controlling MHV-induced pathology and regulation of the immune response. Mice deficient in MDA5 expression (MDA5−/−mice) experienced more severe disease following MHV infection, with reduced survival, increased spread of virus to additional sites of infection, and more extensive liver damage than did wild-type mice. Although type I interferon transcription decreased in MDA5−/−mice, the interferon-stimulated gene response remained intact. Cytokine production by innate and adaptive immune cells was largely intact in MDA5−/−mice, but perforin induction by natural killer cells and levels of interferon gamma, interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α) in serum were elevated. These data suggest that MDA5 signaling reduces the severity of MHV-induced disease, at least in part by reducing the intensity of the proinflammatory cytokine response.

IMPORTANCEMulticellular organisms employ a wide range of sensors to detect viruses and other pathogens. One such sensor, MDA5, detects viral RNA and triggers induction of type I interferons, chemical messengers that induce inflammation and help regulate the immune responses. In this study, we sought to determine the role of MDA5 during infection with mouse hepatitis virus, a murine coronavirus used to model viral hepatitis as well as other human diseases. We found that mice lacking the MDA5 sensor were more susceptible to infection than were mice with MDA5 and experienced decreased survival. Viral replication in the liver was similar in mice with and without MDA5, but liver damage was increased in MDA5−/−mice, suggesting that the immune response is causing the damage. Production of several proinflammatory cytokines was elevated in MDA5−/−mice, suggesting that MDA5 may be responsible for keeping pathological inflammatory responses in check.
Whole-Genome Sequencing of Kaposi's Sarcoma-Associated Herpesvirus from Zambian Kaposi's Sarcoma Biopsy Specimens Reveals Unique Viral Diversity
Authors: Olp Landon N. , Jeanniard Adrien , Marimo Clemence , West John T. , Wood Charles
Published: October 2015

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent for Kaposi's sarcoma (KS). Both KSHV and KS are endemic in sub-Saharan Africa where approximately 84% of global KS cases occur. Nevertheless, whole-genome sequencing of KSHV has only been completed using isolates from Western countries—where KS is not endemic. The lack of whole-genome KSHV sequence data from the most clinically important geographical region, sub-Saharan Africa, represents an important gap since it remains unclear whether genomic diversity has a role on KSHV pathogenesis. We hypothesized that distinct KSHV genotypes might be present in sub-Saharan Africa compared to Western countries. Using a KSHV-targeted enrichment protocol followed by Illumina deep-sequencing, we generated and analyzed 16 unique Zambian, KS-derived, KSHV genomes. We enriched KSHV DNA over cellular DNA 1,851 to 18,235-fold. Enrichment provided coverage levels up to 24,740-fold; therefore, supporting highly confident polymorphism analysis. Multiple alignment of the 16 newly sequenced KSHV genomes showed low level variability across the entire central conserved region. This variability resulted in distinct phylogenetic clustering between Zambian KSHV genomic sequences and those derived from Western countries. Importantly, the phylogenetic segregation of Zambian from Western sequences occurred irrespective of inclusion of the highly variable genes K1 and K15. We also show that four genes within the more conserved region of the KSHV genome contained polymorphisms that partially, but not fully, contributed to the unique Zambian KSHV whole-genome phylogenetic structure. Taken together, our data suggest that the whole KSHV genome should be taken into consideration for accurate viral characterization.

IMPORTANCE
Our results represent the largest number of KSHV whole-genomic sequences published to date and the first time that multiple genomes have been sequenced from sub-Saharan Africa, a geographic area where KS is highly endemic. Based on our new sequence data, it is apparent that whole-genome KSHV diversity is greater than previously appreciated and differential phylogenetic clustering exists between viral genomes of Zambia and Western countries. Furthermore, individual genes may be insufficient for KSHV genetic characterization. Continued investigation of the KSHV genetic landscape is necessary in order to effectively understand the role of viral evolution and sequence diversity on KSHV gene functions and pathogenesis.
Deer Prion Proteins Modulate the Emergence and Adaptation of Chronic Wasting Disease Strains
Authors: Duque Velásquez Camilo , Kim Chiye , Herbst Allen , Daude Nathalie , Garza Maria Carmen , Wille Holger , Aiken Judd , McKenzie Debbie
Published: October 2015

Transmission of chronic wasting disease (CWD) between cervids is influenced by the primary structure of the host cellular prion protein (PrP
C
). In white-tailed deer,
PRNP
alleles encode the polymorphisms Q95 G96 (wild type [wt]), Q95 S96 (referred to as the S96 allele), and H95 G96 (referred to as the H95 allele), which differentially impact CWD progression. We hypothesize that the transmission of CWD prions between deer expressing different allotypes of PrP
C
modifies the contagious agent affecting disease spread. To evaluate the transmission properties of CWD prions derived experimentally from deer of four
PRNP
genotypes (wt/wt, S96/wt, H95/wt, or H95/S96), transgenic (tg) mice expressing the wt allele (tg33) or S96 allele (tg60) were challenged with these prion agents. Passage of deer CWD prions into tg33 mice resulted in 100% attack rates, with the CWD H95/S96 prions having significantly longer incubation periods. The disease signs and neuropathological and protease-resistant prion protein (PrP-res) profiles in infected tg33 mice were similar between groups, indicating that a prion strain (Wisc-1) common to all CWD inocula was amplified. In contrast, tg60 mice developed prion disease only when inoculated with the H95/wt and H95/S96 CWD allotypes. Serial passage in tg60 mice resulted in adaptation of a novel CWD strain (H95
+
) with distinct biological properties. Transmission of first-passage tg60CWD-H95
+
isolates into tg33 mice, however, elicited two prion disease presentations consistent with a mixture of strains associated with different PrP-res glycotypes. Our data indicate that H95-
PRNP
heterozygous deer accumulated two CWD strains whose emergence was dictated by the PrP
C
primary structure of the recipient host. These findings suggest that CWD transmission between cervids expressing distinct PrP
C
molecules results in the generation of novel CWD strains.

IMPORTANCE
CWD prions are contagious among wild and captive cervids in North America and in South Korea. We present data linking the amino acid variant Q95H in white-tailed deer cellular prion protein (PrP
C
) to the emergence of a novel CWD strain (H95
+
). We show that, upon infection, deer expressing H95-PrP
C
molecules accumulated a mixture of CWD strains that selectively propagated depending on the
PRNP
genotype of the host in which they were passaged. Our study also demonstrates that mice expressing the deer S96-
PRNP
allele, previously shown to be resistant to various cervid prions, are susceptible to H95
+
CWD prions. The potential for the generation of novel strains raises the possibility of an expanded host range for CWD.
Induction and Antagonism of Antiviral Responses in Respiratory Syncytial Virus-Infected Pediatric Airway Epithelium
Authors: Villenave Rémi , Broadbent Lindsay , Douglas Isobel , Lyons Jeremy D. , Coyle Peter V. , Teng Michael N. , Tripp Ralph A. , Heaney Liam G. , Shields Michael D. , Power Ultan F.
Published: October 2015

Airway epithelium is the primary target of many respiratory viruses. However, virus induction and antagonism of host responses by human airway epithelium remains poorly understood. To address this, we developed a model of respiratory syncytial virus (RSV) infection based on well-differentiated pediatric primary bronchial epithelial cell cultures (WD-PBECs) that mimics hallmarks of RSV disease in infants. RSV is the most important respiratory viral pathogen in young infants worldwide. We found that RSV induces a potent antiviral state in WD-PBECs that was mediated in part by secreted factors, including interferon lambda 1 (IFN-λ1)/interleukin-29 (IL-29). In contrast, type I IFNs were not detected following RSV infection of WD-PBECs. IFN responses in RSV-infected WD-PBECs reflected those in lower airway samples from RSV-hospitalized infants. In view of the prominence of IL-29, we determined whether recombinant IL-29 treatment of WD-PBECs before or after infection abrogated RSV replication. Interestingly, IL-29 demonstrated prophylactic, but not therapeutic, potential against RSV. The absence of therapeutic potential reflected effective RSV antagonism of IFN-mediated antiviral responses in infected cells. Our data are consistent with RSV nonstructural proteins 1 and/or 2 perturbing the Jak-STAT signaling pathway, with concomitant reduced expression of antiviral effector molecules, such as MxA/B. Antagonism of Jak-STAT signaling was restricted to RSV-infected cells in WD-PBEC cultures. Importantly, our study provides the rationale to further explore IL-29 as a novel RSV prophylactic.

IMPORTANCE
Most respiratory viruses target airway epithelium for infection and replication, which is central to causing disease. However, for most human viruses we have a poor understanding of their interactions with human airway epithelium. Respiratory syncytial virus (RSV) is the most important viral pathogen of young infants. To help understand RSV interactions with pediatric airway epithelium, we previously developed three-dimensional primary cell cultures from infant bronchial epithelium that reproduce several hallmarks of RSV infection in infants, indicating that they represent authentic surrogates of RSV infection in infants. We found that RSV induced a potent antiviral state in these cultures and that a type III interferon, interleukin IL-29 (IL-29), was involved. Indeed, our data suggest that IL-29 has potential to prevent RSV disease. However, we also demonstrated that RSV efficiently circumvents this antiviral immune response and identified mechanisms by which this may occur. Our study provides new insights into RSV interaction with pediatric airway epithelium.
E1a transactivation of human HSP70 gene promoter substitution mutants is independent of the composition of upstream and TATA elements.
Authors: Taylor I C , Kingston R E
Published: October 2015

We have analyzed 41 deletion, linker scan, and substitution mutants of the human HSP70 gene promoter for activation by the adenovirus E1a region. No natural element of the HSP70 gene promoter was required for activation. To investigate specific interactions between E1a and transcription factors, a set of 24 promoters containing all possible combinations of eight different upstream or TATA motifs was investigated for E1a stimulation. E1a transactivated the promoter regardless of the particular TATA motif present. Furthermore, there was no dramatic correlation between any upstream motif and activation by E1a. These data suggest that E1a does not stimulate transcription via an interaction with any specific transcription factor but instead suggest that E1a interacts via the general transcription machinery.
Molecular and functional analysis of the muscle-specific promoter region of the Duchenne muscular dystrophy gene.
Authors: Klamut H J , Gangopadhyay S B , Worton R G , Ray P N
Published: October 2015

Duchenne muscular dystrophy (DMD) gene transcripts are most abundant in normal skeletal and cardiac muscle and accumulate as normal myoblasts differentiate into multinucleated myotubes. In this report we describe our initial studies aimed at defining the cis-acting sequences and trans-acting factors involved in the myogenic regulation of DMD gene transcription. A cosmid clone containing the first exon of the DMD gene has been isolated, and sequences lying upstream of exon 1 were analyzed for homologies to other muscle-specific gene promoters and for their ability to direct muscle-specific transcription of chimeric chloramphenicol acetyltransferase (CAT) gene constructs. The results indicate that the transcriptional start site for this gene lies 37 base pairs (bp) upstream of the 5' end of the published cDNA sequence and that 850 bp of upstream sequence can direct CAT gene expression in a muscle-specific manner. Sequence analysis indicates that in addition to an ATA and GC box, this region contains domains that have been implicated in the regulation of other muscle-specific genes: a CArG box at -91 bp; myocyte-specific enhancer-binding nuclear factor 1 binding site homologies at -58, -535, and -583 bp; and a muscle-CAAT consensus sequence at -394 bp relative to the cap site. Our observation that only 149 bp of upstream sequence is required for muscle-specific expression of a chimeric CAT gene construct further implicates the CArG and myocyte-specific enhancer-binding nuclear factor 1 binding homologies as important domains in the regulation of this gene. On the other hand, the unique profile of myogenic cell line-specific induction displayed by our DMD promoter-CAT gene constructs suggests that other as yet undefined cis-acting sequences and/or trans-acting factors may also be involved.
Regulatory elements mediating transcription from the Drosophila melanogaster actin 5C proximal promoter.
Authors: Chung Y T , Keller E B
Published: October 2015

The major cytoskeletal actin gene of Drosophila melanogaster, the actin 5C gene, has two promoters, the proximal one of which controls constitutive synthesis of actin in all growing tissues. To locate regulatory elements required for constitutive activity of the proximal promoter, mutants of this promoter were fused to the bacterial chloramphenicol acetyltransferase gene and assayed for transient expression activity in cultured Drosophila embryonic Schneider line 2 cells. An essential regulatory element has been located 313 base pairs upstream from the cap site. Deletion of this element lowered expression to one-third of the wild-type level. The element has the sequence AAGTTGTAGTTG, as shown by protein-binding footprinting with the reagent methidiumpropyl-EDTA-Fe(II). This element is probably not a general one, since it was not detected in a search of the published 5'-flanking sequences of 27 Drosophila genes. In addition to this regulatory element, there are five GAGA elements in the actin 5C proximal promoter, some or all of which are essential for the promoter activity as shown by an in vivo competition assay. Although this promoter has no classical TATA element, there is an essential promoter region about 35 base pairs upstream from the cap site that could be a TATA surrogate. The promoter also shows sequences homologous to the alcohol dehydrogenase factor 1-binding site and to the core of the vertebrate serum response element, but mutations of these sites did not affect promoter activity in transient expression assays.
The DNA-binding activity of transcription factor PTF1 parallels the synthesis of pancreas-specific mRNAs during mouse development.
Authors: Petrucco S , Wellauer P K , Hagenbüchle O
Published: October 2015

We have studied the expression of the alpha-amylase, trypsin, and elastase II genes in the acinar pancreas during mouse development. Transcriptional control is the major mechanism by which the differential accumulation of alpha-amylase, trypsin, and elastase II mRNAs is determined during late embryogenesis. The synthesis of pancreatic mRNAs is detected around day 15 of gestation and involves most if not all acinar cells. The DNA-binding activity of the pancreas-specific transcription factor PTF1, which binds to enhancers of genes expressed in this tissue, is detected for the first time at day 15 of gestation. The appearance of the factor at this early stage of development suggests that it plays an important role during pancreas differentiation.
The product of the H19 gene may function as an RNA.
Authors: Brannan C I , Dees E C , Ingram R S , Tilghman S M
Published: October 2015

The mouse H19 gene was identified as an abundant hepatic fetal-specific mRNA under the transcriptional control of a trans-acting locus termed raf. The protein this gene encoded was not apparent from an analysis of its nucleotide sequence, since the mRNA contained multiple translation termination signals in all three reading frames. As a means of assessing which of the 35 small open reading frames might be important to the function of the gene, the human H19 gene was cloned and sequenced. Comparison of the two homologs revealed no conserved open reading frame. Cellular fractionation showed that H19 RNA is cytoplasmic but not associated with the translational machinery. Instead, it is located in a particle with a sedimentation coefficient of approximately 28S. Despite the fact that it is transcribed by RNA polymerase II and is spliced and polyadenylated, we suggest that the H19 RNA is not a classical mRNA. Instead, the product of this unusual gene may be an RNA molecule.
Structure of germ line immunoglobulin alpha heavy-chain RNA and its location on polysomes.
Authors: Radcliffe G , Lin Y C , Julius M , Marcu K B , Stavnezer J
Published: October 2015

We describe the structure of the major germ line RNA transcribed from unrearranged immunoglobulin alpha heavy-chain genes in immunoglobulin M-expressing cells of the I.29 mu B-cell lymphoma, a cell line capable of switching to immunoglobulin A expression upon lipopolysaccharide treatment. This germ line alpha RNA has a small open reading frame that does not include the C alpha domain, and this RNA appears to be present on polysomes in I.29 mu cells.
Transcription of the promoter of the rat NF-1 gene depends on the integrity of an Sp1 recognition site.
Authors: Ammendola R , Gounari F , Piaggio G , De Simone V , Cortese R
Published: October 2015

The transcription start site and promoter of the rat gene coding for the transcription factor NF-1 have been identified. The NF-1 promoter was fused to the chloramphenicol acetyltransferase-coding sequence, and the resulting plasmid was transcriptionally active in the HepG2 cell line. Footprinting and gel retardation analysis indicated that the transcription factor Sp1 binds to the NF-1 promoter. Mutants in the Sp1-binding site displayed a strong reduction in transcriptional activity.
Effects of methotrexate on transfected DNA stability in mammalian cells.
Authors: Pallavicini M G , DeTeresa P S , Rosette C , Gray J W , Wurm F M
Published: October 2015

The chromosomal locations, amounts, and level of expression of transfected, amplified c-myc and dihydrofolate reductase sequences were measured in cells cultured in the presence and absence of methotrexate. These studies show that the location and amount of transfected sequences, as well as the level of expression, were more variable when the cells were cultured in methotrexate.
A gene that encodes a protein consisting solely of zinc finger domains is preferentially expressed in transformed mouse cells.
Authors: Ernoult-Lange M , Kress M , Hamer D
Published: October 2015

We describe the cloning and characterization of the mouse MOK-2 gene, a new member of the Krüppel family of zinc finger proteins. Sequencing of both cDNA and genomic clones showed that the predicted MOK-2 protein consists of seven zinc finger domains with only five additional amino acids. The finger domains of MOK-2 are highly homologous to one another but not to those of other zinc finger proteins. MOK-2 is preferentially expressed in transformed cell lines, brain tissue, and testis tissue. Its possible role in cellular transformation is discussed.