bluuebunny/crossref_metadata_2025 huggingface dataset:
Administration of Modified Spherosome Suspension (BY963) Leads to an Increase of Acoustic Impedance in Dog Brain Tissue
Authors: Seidel G. , Beller K-D. , Kaps M.
Published: February 2016

Ultrasound contrast agents change the acoustic properties of brain tissue. This can be quantified with acoustic densitometry. In a dog model, the authors examined changes in acoustic impedance in the thalamic and parietal white‐matter regions of the brain after intravenous injection of the spherosome containing an ultrasound contrast agent (BY963) filled with perfluoropentane gas. The authors examined six sedated mongrel dogs with a Hewlett‐Packard Sonos 1500 device. BY963 filled with perfluoropentane (0.2 ml/kg body weight) was injected three times with a time interval between injections of 5 minutes. Time‐dependent changes in mean acoustic impedance were calculated. The authors found a significant increase in peak acoustic impedance after fractional injection of 0.6 ml/kg body weight (3 × 0.2 ml/kg body weight) in the thalamus region up to 7.0 IU (p = 0.006). In the parietal white matter the increase in peak acoustic impedance was not significant (p = 0.06). Statistical comparison of the increase in peak acoustic impedance between placebo and BY963 injection in the thalamus region showed a significant difference after the first injection (p = 0.01) but showed no significance after the second and third injections. The authors concluded that thalamus and parietal white matter of the brain showed different accumulations of BY963.
Mirror‐Image Artifact Can Affect Transcranial Doppler Interpretation
Authors: Ratanakorn Disya , Kremkau Frederick W. , Myers Lawrence G. , Meads Dana B. , Tegeler Charles H.
Published: February 2016

Transcranial Doppler ultrasonography (TCD) allows evaluation of blood‐flow velocity in intracranial arteries detection and monitoring of vasospasm in patients with subarachnoid hemorrhage. Spectral Doppler artifacts can affect TCD data. A 1‐month series of TCD findings showed marked fluctuation in blood‐flow velocity values in both the middle and anterior cerebral arteries of a patient with subarachnoid hemorrhage. A mirror‐image artifact of the Doppler fast Fourier transform velocity spectrum resulted in erroneous interpretation of higher flow velocity in certain vessels. This artifact may cause misinterpretation of TCD flow‐velocity data and lead to improper diagnosis of the condition and treatment of patients.
A 67‐Year‐Old Woman with Polymyalgia Rheumatica and Left Hemispatial Neglect
Authors: Mahfood J. Paul , Gold Michael , Gonzalvo America , Valeriano-Marcet Joanne
Published: February 2016

A 67‐year‐old woman with a diagnosis of polymyalgia rheumatica presented initially wih periods of confusion and incontinence ACT scan of the brain was normal and she was treated with tapering doses of corticosteroids and clinical improvement After a brief period off steroids, the patient presented with a progressive dementia, left‐sided clumsiness, gait disturbances and left hemispatial neglect An MRI at this time demonstrated a large area of edema over the right parietal lobe and intense cortical enhancement A chest CT demonstrated multiple nodules Biopsies of the lung and brain failed to identify any infectious organisms or malignant tissue The leptomeningeal biopsy revealed multiple granulomatous areas with central necrosis and hystiocytic cells consistent with idiopathic hypertrophic pachymeningitis
Intima‐media Thickness of Carotid Arteries in Borderline Hypertensives
Authors: Pit'ha J , Krajičková D , Cífková R , Hubáček J , Petržilková Z , Hejl Z , Stávek P , Skibová J , Poledne R
Published: February 2016

The purpose of this study was to assess the effect of the main risk factors for cardiovascular disease on the process of subclinical atherosclerosis in originally borderline hypertensives. The relation of far wall common carotid artery intima‐media thickness (IMT CCA) measured by B‐mode ultrasound to smoking, body mass index (BMI), blood pressure, lipids, and angiotensin‐converting enzyme (ACE) gene polymorphism was analyzed. In 48 subjects examined (mean age, 61.9 ± 2.54 years), median IMT CCA was 0.708 mm. Statistically significant differences in BMI (26.5 vs. 29.2 kg/m2, p<0.025) and HDL‐cholesterol level (1.42 vs. 1.1 mmol/l, p<0.025) between the first and third tertile of IMT CCA were found. No differences were observed between “controls” and “cases” in blood pressure, total cholesterol, and triacylglycerols. No significant differences in IMT CCA were found between smokers and nonsmokers and among different alleles of the ACE gene. These data reflect the importance of HDL‐cholesterol and BMI on the process of atherosclerosis within an otherwise homogeneous group of patients.
Clinical Application of Magnetoencephalography in a Patient with Corticobasal Degeneration
Authors: Mizuno Toshiki , Takanashi Yoshiaki , Nakase Taizen , Makino Masahiro , Iwamoto Kazuhide , Nakajima Kenji , Furuya Seiichi , Naruse Shoji , Imahori Yoshio , Yoshida Yoshikazu
Published: February 2016

Previous neuroimaging studies in corticobasal degeneration (CBD), such as MRI and positron emission tomography, showed lateralized cortical atrophy and a decrease in cortical oxygen metabolism, respectively. The advantage of magnetoencephalography in the research of the auditory system is that activity of both hemispheres can be detected separately. Auditory‐evoked magnetic fields (AEFs) were then applied to a patient with corticobasal degeneration. The strength of N1 OOm equivalent current dipoles (ECD) in left hemisphere was extremely smaller (20 nAm) than that in right hemisphere (48 nAm). This results implies that CBD patient might have a decrease in the number of neurons in the primary auditory cortex. This study suggests that AEF measurement is one of the most powerful tools for detecting latent impairment of auditory function in patients with corticobasal degeneration.
Brain Uptake and Plasma Metabolism of [11C]Vinpocetine: A Preliminary PET Study in a Cynomolgus Monkey
Authors: Gulyás Balázs , Halldin Christer , Karlsson Per , Chou Yuan-H. , Swahn Cari-Gunnar , Bönöck Peter , Paróczai Margit , Farde Lars
Published: February 2016

Vinpocetine, a vinca alkaloid, is a widely used therapeutic agent in patients with acute and chronic stroke. To reveal the mechanisms of vinpocetine action in the brain, vinpocetine was labeled with 11C. Positron emission tomography (PET) was used to determine the uptake and distribution of [11C]vinpocetine in brain regions and the trunk of a cynomolgous monkey in two independent measurements. The concentration of vinpocetine and its labeled metabolites was determined in blood and plasma using high‐performance liquid chromatography (HPLC). Almost identical measurements were obtained in the two independent studies. After intravenous administration, following an initial peak, the total concentration of radioactivity in blood was relatively stable with time, whereas the concentration of the unchanged compound decreased with time in an exponential manner. The uptake of [11C]vinpocetine in brain was rapid, and 5% of the radioactivity totally injected was present in the brain 2 minutes after drug administration, indicating that the compound entered the brain readily. The radioactivity uptake was heterogeneously distributed among brain regions and was highest in the thalamus, the basal ganglia, and certain neocortical regions. The high brain uptake and the heterogeneous regional distribution indicate that direct central nervous system (CNS) effects of vinpocetine must be considered as explanation for the therapeutic effects. The detailed exploration of this suggestion requires further studies.
Basilar Artery Dissection in a Young Woman: A Case Report
Authors: Kothari Milind J. , Stabley Jason , Wojnar Margaret , Edgar Kenneth , Jones Blaise V. , McNamara Kevin
Published: February 2016

The case of a young woman with basilar artery dissection, possibly precipitated by trauma, is presented, and the literature is reviewed.
Hemispheric Language Dominance Testing by Means of fMRI
Authors: Baciu Monica , Rubin Christophe , Décorps Michel , Segebarth Christoph
Published: February 2016

Presurgical evaluation of patients with intractable epilepsy routinely includes hemispheric language dominance (HLD) testing. Commonly, HLD is assessed by the intracarotid amobarbital (Wada) test.1 The advent of functional magnetic resonance imaging (fMRI) has generated considerable interest in this technique as an alternative to HLD testing.2–6 Functional MRI mapping of the brain relies on the detection of focal changes in the concentrations of deoxyhemoglobin.7
Blood Flow Velocities in Middle Cerebral Artery Branches After Subarachnoid Hemorrhage
Authors: Mursch Kay , Bransi Ahmad , Vatter Hartmut , Herrendorf Gregor , Behnke‐Mursch Julianne , Kolenda Herbert
Published: February 2016

In a prospective study, 55 patients were examined by transcranial duplex sonography (TCCS) after subarachnoid hemorrhage (SAH) to determine whether additional transcranial duplex examination on the middle cerebral artery M2 segments would aid in the examination of the MCA stem segment. The mean blood flow velocities and pulsatility index were correlated to the occurrence of delayed ischemic neurologic deficits (DIND). Out of 47 patients included, 21 did not experience any delayed deficit (group 1), 15 did (group II), and in 11 the extent to which vasospasm contributed to a neurologic deficit was unclear (group III). The highest blood flow velocity and the greatest increase of mean blood flow velocity on 1 day were significantly higher in groups II and III both in M1 and in M2. In 10 patients in group II, where the onset day of DIND was known exactly, Doppler data indicating ischemia before or at the time of DIND were observed in nine. In eight patients, Doppler of the MCA stem alone would have provided enough information to recognize the risk of symptomatic vasospasm; in one patient, only the M2 Doppler gave an indication of ischemic complication. Transcranial duplex sonography may provide additional information to TCD by accurate delineation of M1/M2 vasospasm and therefore may help plan cerebral angiography and neurointerventional treatment.
Serial Transcranial Color‐Coded Sonography in Postpartum Cerebral Angiopathy
Authors: Ihara Masafumi , Yanagihara Chie , Nishimura Yo
Published: February 2016

Reversible vasospasm in the bilateral middle cerebral artery in a patient with postpartum cerebral angiopathy was evaluated with serial transcranial color‐coded sonography (TCCS). The authors propose TCCS as the method of choice for assessing the time course of vascular changes in postpartum cerebral angiopathy because it allows for precise placement of the sample volume and adjustment of the incident angle.
THBS1 is induced by TGFB1 in the cancer stroma and promotes invasion of oral squamous cell carcinoma
Authors: Pal Samir Kumar , Nguyen Chi Thi Kim , Morita Kei‐ichi , Miki Yoshio , Kayamori Kou , Yamaguchi Akira , Sakamoto Kei
Published: February 2016

Background

THBS1 (thrombospondin‐1) is the extracellular matrix (ECM) protein that affects diverse cellular activities. It constitutes the tumor stroma, but the role of THBS1 in oral squamous cell carcinoma (OSCC) development is unclear. The aim of this study was to clarify the relevance of THBS1 in the pathogenesis of OSCC.

Materials and methods

The expression of THBS1 was examined in 44 OSCC by immunohistochemical analysis and in 43 OSCC by cDNA microarray analysis. Cell culture experiments were conducted using human OSCC cell lines HSC3 and HO1N1 and mouse fibroblast ST2 cells to examine the effect of TGFB1 on THBS1 expression, and the effect of THBS1 on cellular behaviors.

Results

THBS1 was specifically induced in the tumor microenvironment of OSCC. THBS1 appeared to be produced mainly by the stromal cells, but also by OSCC cells. TGFB1 stimulated THBS1 expression in ST2, primary fibroblasts, and the OSCC cells. THBS1 promoted migration and invasion of HSC3 and HO1N1 in transwell migration assays. THBS1 stimulated the expression of MMP3 (matrix metalloprotease 3), MMP9, MMP11, and MMP13 in ST2 cells and MMP3, MMP11, and MMP13 in HO1N1 cells. The RGD peptide suppressed the THBS1‐stimulated migration and upregulation of MMP11 and MMP13.

Conclusions

THBS1 is a tumor‐specific ECM protein that is induced by TGFB1 and promotes migration of cancer cells and stimulates the expression of MMPs partly through the integrin signaling, thereby favoring OSCC invasion.
Comparison of Maximum Intercuspal Contacts of Articulated Casts and Virtual Casts Requiring Posterior Fixed Partial Dentures
Authors: Arslan Yeliz , Bankoğlu Güngör Merve , Karakoca Nemli Seçil , Kökdoğan Boyacı Betül , Aydın Cemal
Published: February 2016

Purpose

To evaluate the accuracy of the CEREC CAD/CAM system in reproducing the maximum intercuspal contacts of the casts, which include posterior teeth preparation for a fixed partial denture (FPD).

Materials and Methods

Ten pairs of gypsum casts were mounted in articulators in maximum intercuspal position (MIP) to serve as patient simulation models. Tooth #19 was removed from the cast. Occlusal contacts in MIP were identified with articulating paper, and digital impressions of the casts with unprepared teeth and buccal images in MIP were taken. Teeth #18 and #20 were prepared for an FPD, and full‐ and half‐arch digital impressions of the casts with prepared teeth and buccal images from different sides were taken. In each situation, screenshot images of the virtual casts with occlusal contacts were saved as JPEG files. The proportions of congruence of virtual contacts with cast contacts were analyzed by superimposing screenshot images of the virtual casts onto the screenshot images of the casts with the indicated occlusal contacts in a transparent manner using an image‐processing program. The data were statistically analyzed with a paired t‐test.

Results

The highest percentages of virtually indicated contacts identical to the cast contacts were observed in non‐prepared full‐arch digital impressions. Comparison of full‐arch impressions taken before and after tooth preparation showed no difference for congruence even if the buccal image was taken from the contralateral or ipsilateral side (p > 0.05). After tooth preparation, comparing full‐ and half‐arch digital impressions revealed that half‐arch impression showed significantly lower percentages of identical contacts (p < 0.05). When comparing the buccal image side, no significant difference was detected between ipsilateral and contralateral images both for non‐prepared and prepared casts (p > 0.05).

Conclusion

When there is no posterior antagonist contact following tooth preparation for an FPD, taking a full‐arch digital impression and designing the restoration on full‐arch virtual models can be advocated.
Influence of Veneering Fabrication Techniques and Gas‐Phase Fluorination on Bond Strength between Zirconia and Veneering Ceramics
Authors: Pharr Stewart W. , Teixeira Erica C. , Verrett Ronald , Piascik Jeffrey R.
Published: February 2016

Purpose

Porcelain chipping has been one of the main problems of porcelain‐fused‐to‐zirconia restorations. This study evaluates the bond strengths of layered, pressed, and adhesively bonded porcelain to yttria‐stabilized zirconia substrates that have undergone traditional preparation or gas‐phase fluorination.

Materials and Methods

A three‐point bending test was used to evaluate the bond strength of the porcelain and zirconia interface. Sixty‐six specimens were prepared (n = 11) following ISO 9693 and loaded until failure using an Instron testing machine. One‐half of the zirconia substrates received gas phase fluorination treatment before veneering application. Three porcelain veneering methods were evaluated: layered, pressed, and adhesively bonded porcelain. Bond strength results were interpreted using a two‐way ANOVA and a Bonferroni multiple comparisons test. Statistical significance was set at α = 0.05.

Results

ANOVA revealed a statistically significant effect of the veneering fabrication methods. No main effect was observed regarding the surface treatment to the zirconia. There was a significant effect related to the veneering method used to apply porcelain to zirconia. For untreated zirconia, layered porcelain had a significantly higher flexural strength compared to pressed or bonded, while pressed and bonded porcelains were not significantly different from one another. For zirconia specimens receiving fluorination treatment, both layered and pressed porcelains had significantly higher bond strengths than adhesively bonded porcelain. In addition, fluorinated pressed porcelain was not statistically different from the control layered or fluorinated layered porcelain.

Conclusion

The choice of veneering fabrication technique was critical when evaluating the zirconia to porcelain interfacial bond strength. Bonded porcelain to zirconia had a lower flexural strength than layered or pressed porcelain, regardless of zirconia surface treatment. In addition, fluorination had an effect on the bond strength of pressed porcelain.
Whose “Storm and Stress” Is It? Parent and Child Reports of Personality Development in the Transition to Early Adolescence
Authors: Göllner Richard , Roberts Brent W. , Damian Rodica I. , Lüdtke Oliver , Jonkmann Kathrin , Trautwein Ulrich
Published: February 2016

The present study investigated Big Five personality trait development in the transition to early adolescence (from the fifth to eighth grade). Personality traits were assessed in 2,761 (47% female) students over a 3‐year period of time. Youths’ self‐reports and parent ratings were used to test for cross‐informant agreement. Acquiescent responding and measurement invariance were established with latent variable modeling. Growth curve models revealed three main findings: (a) Normative mean‐level changes occurred for youths’ self‐report data and parent ratings with modest effects in both cases. (b) Agreeableness and Openness decreased for self‐reports and parent ratings, whereas data source differences were found for Conscientiousness (decreased for self‐reports and remained stable for parent ratings), Extraversion (increased for self‐reports and decreased for parent ratings), and Neuroticism (remained stable for self‐reports and decreased for parent ratings). (c) Girls showed a more mature personality overall (self‐reports and parent ratings revealed higher levels of Agreeableness, Conscientiousness, and Openness) and became more extraverted in the middle of adolescence (self‐reports). Personality changes modestly during early adolescence whereby change does not occur in the direction of maturation, and substantial differences exist between parent ratings and self‐reports.
Can breeder reproductive status, performance and egg quality be enhanced by supplementation and transition of n‐3 fatty acids?
Authors: Delezie E. , Koppenol A. , Buyse J. , Everaert N.
Published: February 2016

The aim of this experiment was to investigate the effect of n‐3 fatty acid (FA) supplemented diets on breeder performance, productivity and egg quality. Breeders (n = 480) were fed the supplemented diet from 18 weeks onwards; the inclusion level of n‐3 FA was increased from 1.5% to 3.0% from 34 weeks of age onwards until 48 weeks of age. Ross‐308 broiler breeders (n = 480) were fed one of four different diets: a basal diet rich in n‐6 FA (control diet) or one of three diets rich in n‐3 FA. For the n‐3 FA diets, eicosapentaenoic acid (EPA, 20:5 n‐3) and docosahexaenoic acid (DHA, 22:6 n‐3) were fed to the broiler breeders at different ratios formulated to obtain EPA/DHA ratios of 1/1, 1/2 or 2/1. Differences in performance, reproduction and egg quality parameters due to n‐3 supplementation were noted more for the 1.5% followed by the 3.0% fed broilers than their 1.5% supplemented counterparts. Egg weight (p < 0.001) and egg mass (p = 0.003) were significantly lower and feed conversion (p = 0.008) significantly higher for the n‐3 FA (at 3.0% inclusion level) fed broilers compared to the control group. For the EPA‐ and DHA‐fed breeders, a higher proportional abdominal fat percentage (p = 0.025) and proportional albumen weight (%) (p = 0.041) were found respectively. Dietary treatments did not affect reproduction. It can be concluded that the results of the present experiment indicate no significant differences between treatments at 1.5% inclusion levels. However, increasing this level to 3.0% is not recommended due to the rather negative effects on the measured parameters. It should be further investigated whether these adverse effects were obtained due to (i) the higher supplementation level, (ii) combining a supplementation level of 1.5% with 3% or (iii) the duration of supplementation.
A genetic delineation of Patchouli (Pogostemon cablin) revealed by specific‐locus amplified fragment sequencing
Authors: Huang Hui‐Run , Wu Wei , Zhang Ji‐Xiu , Wang Ling‐Jian , Yuan Yong‐Ming , Ge Xue‐Jun
Published: February 2016

Patchouli plant (Pogostemon cablin (Blanco) Benth.) is of important economic value, and it has been grown for medicinal use for more than 1000 years in China and Southeast Asia. There are limited data to underpin the genetic and genomic resource management for Patchouli. Herein, we used specific‐locus amplified fragment sequencing to generate a genetic delineation of P. cablin collected from Vietnam, South China, and Indonesia (Sumatra). In total, 15 457 835 reads, 61 334 specific‐locus amplified fragments, and 511 reliable single nucleotide polymorphisms were obtained. On the basis of model‐based grouping and neighbor‐joining trees, we divided the studied accessions into six distinct groups: one Vietnamese group, two Chinese groups, and three Indonesian groups. We also measured the contents of patchouli alcohol and pogostone; all accessions belonged to patchoulol‐type except for three accessions from the species' northernmost distribution in China, which have a high content of pogostone. The results from both genetic structure and chemotypes were highly consistent with the possible migration history of Patchouli. Accordingly, ex situ conservation should be immediately established for Patchouli, particularly for the pogostone‐type and the germplasm in Vietnam.
The multivariate association between genomewide DNA methylation and climate across the range of Arabidopsis thaliana
Authors: Keller Thomas E. , Lasky Jesse R. , Yi Soojin V.
Published: February 2016

Epigenetic changes can occur due to extracellular environmental conditions. Consequently, epigenetic mechanisms can play an intermediate role to translate environmental signals to intracellular changes. Such a role might be particularly important in plants, which often show strong local adaptation and have the potential for heritable epigenetic states. However, little is currently known about the role of epigenetic variation in the ecological mechanisms of adaptation. Here, we used multivariate redundancy analyses to examine genomewide associations between DNA methylation polymorphisms and climate variation in two independent panels of Arabidopsis accessions, including 122 Eurasian accessions as well as in a regional panel of 148 accessions in Sweden. At the single‐nucleotide methylation level, climate and space (geographic spatial structure) explain small yet significant amount of variation in both panels. On the other hand, when viewed in a context of genomic clusters of methylated and unmethylated cytosines, climate and space variables explain much greater amounts of variation in DNA methylation than those explained by variation at the single‐nucleotide level. We found that the single‐nucleotide methylation polymorphisms with the strongest associations with climate were enriched in transposable elements and in potentially RNA‐directed methylation contexts. When viewed in the context of genomic clusters, variation of DNA methylation at different sequence contexts exhibit distinctive segregation along different axes of variation in the redundancy analyses. Genomewide methylation showed much stronger associations with climate within the regional panel (Sweden) compared to the global (Eurasia). Together, these findings indicate that genetic and epigenetic variation across the genome may play a role in response to climate conditions and local adaptation.
Hybridization during altitudinal range shifts: nuclear introgression leads to extensive cyto‐nuclear discordance in the fire salamander
Authors: Pereira Ricardo J. , Martínez‐Solano Iñigo , Buckley David
Published: February 2016

Ecological models predict that, in the face of climate change, taxa occupying steep altitudinal gradients will shift their distributions, leading to the contraction or extinction of the high‐elevation (cold‐adapted) taxa. However, hybridization between ecomorphologically divergent taxa commonly occurs in nature and may lead to alternative evolutionary outcomes, such as genetic merger or gene flow at specific genes. We evaluate this hypothesis by studying patterns of divergence and gene flow across three replicate contact zones between high‐ and low‐elevation ecomorphs of the fire salamander (Salamandra salamandra) that have experienced altitudinal range shifts over the current postglacial period. Strong population structure with high genetic divergence in mitochondrial DNA suggests that vicariant evolution has occurred over several glacial–interglacial cycles and that it has led to cryptic differentiation within ecomorphs. In current parapatric boundaries, we do not find evidence for local extinction and replacement upon postglacial expansion. Instead, parapatric taxa recurrently show discordance between mitochondrial and nuclear markers, suggesting nuclear‐mediated gene flow across contact zones. Isolation with migration models support this hypothesis by showing significant gene flow across all five parapatric boundaries. Together, our results suggest that, while some genomic regions, such as the mitochondria, may follow morphologic species traits and retreat to isolated mountain tops, other genomic regions, such as nuclear markers, may flow across parapatric boundaries, sometimes leading to a complete genetic merger. We show that despite high ecologic and morphologic divergence over prolonged periods of time, hybridization allows for evolutionary outcomes alternative to extinction and replacement of taxa in response to climate change.
Comparative spatial genetics and epigenetics of plant populations: heuristic value and a proof of concept
Authors: Herrera Carlos M. , Medrano Mónica , Bazaga Pilar
Published: February 2016

Despite the recent upsurge of interest on natural epigenetic variation of nonmodel organisms, factors conditioning the spatial structure of epigenetic diversity in wild plant populations remain virtually unexplored. We propose that information on processes shaping natural epigenetic variation can be gained using the spatial structure of genetic diversity as null model. Departures of epigenetic isolation‐by‐distance (IBD) patterns from genetic IBD patterns for the same sample, particularly differences in slope of similarity‐distance regressions, will reflect the action of factors that operate specifically on epigenetic variation, including imperfect transgenerational inheritance and responsiveness to environmental factors of epigenetic marks. As a proof of concept, we provide a comparative analysis of spatial genetic and epigenetic structure of 200 mapped individuals of the perennial herb Helleborus foetidus. Plants were fingerprinted using nuclear microsatellites, amplified fragment length polymorphisms (AFLP) and methylation‐sensitive AFLP markers. Expectations from individual‐level IBD patterns were tested by means of kinship‐distance regressions. Both genetic and epigenetic similarity between H. foetidus individuals conformed to theoretical expectations under individual‐level IBD models. Irrespective of marker type, there were significant negative linear relationships between the kinship coefficient for plant pairs and their spatial separation. Regression slopes were significantly steeper for epigenetic markers. Epigenetic similarity between individuals was much greater than genetic similarity at shortest distances, such epigenetic ‘kinship excess’ tending to decrease as plant separation increased. Results suggest that moderate‐to‐high heritability and responsiveness to local environments are major drivers of epigenetic spatial structure in H. foetidus, and illustrate the heuristic value of comparing genetic and epigenetic spatial structure for formulating and testing hypotheses on forces shaping epigenetic diversity in wild plant populations.
Development and evaluation of real‐time PCR assays for bloodmeal identification in Culicoides midges
Authors: VAN DER SAAG M. R. , GU X. , WARD M. P. , KIRKLAND P. D.
Published: February 2016

Culicoides (Diptera: Ceratopogonidae) midges are the biological vectors of a number of arboviruses of veterinary importance. However, knowledge relating to the basic biology of some species, including their host‐feeding preferences, is limited. Identification of host‐feeding preferences in haematophagous insects can help to elucidate the transmission dynamics of the arboviruses they may transmit. In this study, a series of semi‐quantitative real‐time polymerase chain reaction (qPCR) assays to identify the vertebrate host sources of bloodmeals of Culicoides midges was developed. Two pan‐reactive species group and seven species‐specific qPCR assays were developed and evaluated. The assays are quick to perform and less expensive than nucleic acid sequencing of bloodmeals. Using these assays, it was possible to rapidly test nearly 700 blood‐fed midges of various species from several geographic locations in Australia.
When virulence originates from nonagricultural hosts: evolutionary and epidemiological consequences of introgressions following secondary contacts inVenturia inaequalis
Authors: Leroy Thibault , Caffier Valérie , Celton Jean‐Marc , Anger Nicolas , Durel Charles‐Eric , Lemaire Christophe , Le Cam Bruno
Published: February 2016

In pathogens, introgressions through secondary contacts between divergent populations from agricultural and nonagricultural disease reservoirs are expected to have crucial evolutionary and epidemiological implications. Despite the importance of this question for disease management, experimental demonstrations of these implications remain scarce.

Recently, we identified a virulent population of the apple scab pathogenVenturia inaequalisthat migrated from nonagricultural hosts to European domestic apple orchards. Here, we investigated the occurrence of gene flow between agricultural and nonagricultural populations sampled in two orchards, and thereafter its consequences on the pathogenicity of hybrids.

Population genetic structure and demographic inferences based on the genotypes of 104 strains revealed a high amount of gene flow between the two populations in one orchard. In this site, mating between populations was made possible by the presence of a common host. Our results revealed an invasion of the virulent trait in the agricultural population; a main direction of introgression in hybrids from the agricultural to nonagricultural genetic backgrounds; and a population of hybrids with transgressive traits.

We demonstrate a secondary contact with gene flow between divergent populations of pathogens. Our findings highlight evolutionary and epidemiological changes in pathogens and have concrete implications for sustainable disease management.
Noninvasive brain stimulation treatments for addiction and major depression
Authors: Dunlop Katharine , Hanlon Colleen A. , Downar Jonathan
Published: February 2016

Major depressive disorder (MDD) and substance use disorders (SUDs) are prevalent, disabling, and challenging illnesses for which new treatment options are needed, particularly in comorbid cases. Neuroimaging studies of the functional architecture of the brain suggest common neural substrates underlying MDD and SUDs. Intrinsic brain activity is organized into a set of functional networks, of which two are particularly relevant to psychiatry. The salience network (SN) is crucial for cognitive control and response inhibition, and deficits in SN function are implicated across a wide variety of psychiatric disorders, including MDD and SUDs. The ventromedial network (VMN) corresponds to the classic reward circuit, and pathological VMN activity for drug cues/negative stimuli is seen in SUDs/MDD. Noninvasive brain stimulation (NIBS) techniques, including rTMS and tDCS, have been used to enhance cortico–striatal–thalamic activity through the core SN nodes in the dorsal anterior cingulate cortex, dorsolateral prefrontal cortex, and anterior insula. Improvements in both MDD and SUD symptoms ensue, including in comorbid cases, via enhanced cognitive control. Inhibition of the VMN also appears promising in preclinical studies for quenching the pathological incentive salience underlying SUDs and MDD. Evolving techniques may further enhance the efficacy of NIBS for MDD and SUD cases that are unresponsive to conventional treatments.
Enhancing the evaluation of PI3K inhibitors through 3D melanoma models
Authors: Shannan Batool , Chen Quan , Watters Andrea , Perego Michela , Krepler Clemens , Thombre Rakhee , Li Ling , Rajan Geena , Peterson Scott , Gimotty Phyllis A. , Wilson Melissa , Nathanson Katherine L. , Gangadhar Tara C. , Schuchter Lynn M. , Weeraratna Ashani T. , Herlyn Meenhard , Vultur Adina
Published: February 2016

Targeted therapies for mutant BRAF metastatic melanoma are effective but not curative due to acquisition of resistance. PI3K signaling is a common mediator of therapy resistance in melanoma; thus, the need for effective PI3K inhibitors is critical. However, testing PI3K inhibitors in adherent cultures is not always reflective of their potential in vivo. To emphasize this, we compared PI3K inhibitors of different specificity in two‐ and three‐dimensional (2D, 3D) melanoma models and show that drug response predictions gain from evaluation using 3D models. Our results in 3D demonstrate the anti‐invasive potential of PI3K inhibitors and that drugs such as PX‐866 have beneficial activity in physiological models alone and when combined with BRAF inhibition. These assays finally help highlight pathway effectors that could be involved in drug response in different environments (e.g. p4E‐BP1). Our findings show the advantages of 3D melanoma models to enhance our understanding of PI3K inhibitors.
Epidemiological Evidence for a Decreasing Incidence of Neonatal Abstinence Syndrome, 2000–11
Authors: Uebel Hannah , Wright Ian M. , Burns Lucy , Hilder Lisa , Bajuk Barbara , Breen Courtney , Abdel‐Latif Mohamed E. , Ward Meredith , Eastwood John , Feller John M. , Falconer Janet , Clews Sara , Oei Ju Lee
Published: February 2016

Background

This study analyses the incidence of Neonatal Abstinence Syndrome (NAS) in a large geographically defined population in Australia.

Method

Database linkage analysis of all births between 2000 and 2011 in New South Wales (NSW), Australia. The diagnosis of NAS was derived from hospital coding P96.1, ‘Neonatal withdrawal symptoms from maternal use of drugs of addiction’. Temporal trends were studied by comparing epoch 1 (2000–05) with epoch 2 (2006–11). The relationship with changes in maternal factors was further analysed.

Results

The NAS was coded in 3842 of 1 022 263 live born infants (0.38%). NAS incidence peaked at 5.07 per 1000 live births in 2002, decreasing to 3.18 in 2011 and was negatively correlated with maternal age (r = −0.7). The rate of NAS in epoch 2 (3.4 per 1000 births, 95% CI 3.28, 3.58) was significantly lower than in epoch 1 (4.1 per 1000 births, 95% CI 3.96, 4.33). Epoch 2 mothers were significantly older (mean 29.8 years vs. 28.3 years), less likely to be multiparous (OR 0.7, 95% CI 0.6, 0.9) or smoke (OR 0.4, 95% CI 0.4, 0.5). They were more likely to engage in antenatal care earlier (mean first visit: 14.1 vs. 18.9 weeks). Most infants (~80%) were born at term (>37 weeks gestation).

Conclusion

The incidence of NAS as a discharge diagnosis has decreased in our population since 2002. Mothers are also older and engaging earlier in prenatal care. Whether these changes alter NAS presentation and diagnosis or whether pregnant women are using drugs that do not cause typical NAS (e.g. amphetamines) is uncertain and requires further study.
Regulatory Transformation: Lessons from Connecticut's Department of Energy and Environmental Protection
Authors: Esty Daniel
Published: February 2016

As economies and citizen priorities continue to evolve, governance practices must also adjust to changing circumstances and public expectations. This article explores the important topic of regulatory transformation, drawing from both the academic literature and the author's recent experience heading Connecticut's Department of Energy and Environmental Protection. The article makes the case for a focus on vision and execution as the critical starting points for good governance in the twenty‐first century, drawing on best practices from the world of management. It then addresses some of the constraints faced by government organizations in this evolving era. Finally, the article identifies five core elements of regulatory excellence: integration, innovation, incentives, investment, and implementation.
Taylor's Power Law and the Statistical Modelling of Infectious Disease Surveillance Data
Authors: Enki Doyo Gragn , Noufaily Angela , Farrington Paddy , Garthwaite Paul , Andrews Nick , Charlett Andre
Published: February 2016

Surveillance data collected on several hundred different infectious organisms over 20 years have revealed striking power relationships between their variance and mean in successive time periods. Such patterns are common in ecology, where they are referred to collectively as Taylor's power law. In the paper, these relationships are investigated in detail, with the aim of exploiting them for the descriptive statistical modelling of infectious disease surveillance data. We confirm the existence of variance-to-mean power relationships, with exponent typically between 1 and 2. We investigate skewness-to-mean relationships, which are found broadly to match those expected of Tweedie distributions, and thus confirm the relevance of the Tweedie convergence theorem in this context. We suggest that variance- and skewness-to-mean power laws, when present, should inform statistical modelling of infectious disease surveillance data, notably in descriptive analysis, model building, simulation and interval and threshold estimation, threshold estimation being particularly relevant to outbreak detection.
Identifying Subgroups of Enhanced Predictive Accuracy from Longitudinal Biomarker Data by Using Tree-Based Approaches: Applications to Fetal Growth
Authors: Foster Jared C. , Liu Danping , Albert Paul S. , Liu Aiyi
Published: February 2016

Longitudinal monitoring of biomarkers is often helpful for predicting disease or a poor clinical outcome. We consider the prediction of both large and small for gestational age births by using longitudinal ultrasound measurements, and we attempt to identify subgroups of women for whom prediction is more (or less) accurate, should they exist. We propose a tree-based approach to identifying such subgroups, and a pruning algorithm which explicitly incorporates a desired type I error rate, allowing us to control the risk of false discovery of subgroups. The methods proposed are applied to data from the Scandinavian Fetal Growth Study and are evaluated via simulations.
Assessing the Inequality of Lifetime Healthcare Expenditures: a Nearest Neighbour Resampling Approach
Authors: Wong Albert , Boshuizen Hendriek , Polder Johan , Ferreira José António
Published: February 2016

The rise in healthcare expenditures has raised doubts about the sustainability of health systems and instigated a discussion on their design. Policy making in this field requires a proper understanding of how healthcare expenditures evolve throughout an individual's lifetime, and of how they vary between individuals. Given the lack of data on healthcare expenditures during an individual's lifetime, we developed a new nearest neighbour resampling approach to construct realistic individual life cycles of healthcare expenditures based on cross-sectional data from the Netherlands. This approach provides insight into lifetime healthcare expenditures. Our main finding is that the inequality in lifetime healthcare expenditures is much smaller than the inequality as derived from cross-sectional healthcare expenditures.
A TILLING approach to generate broad‐spectrum resistance to potyviruses in tomato is hampered by eIF4E gene redundancy
Authors: Gauffier Camille , Lebaron Caroline , Moretti André , Constant Carole , Moquet Frédéric , Bonnet Grégori , Caranta Carole , Gallois Jean‐Luc
Published: February 2016

Genetic resistance to pathogens is important for sustainable maintenance of crop yields. Recent biotechnologies offer alternative approaches to generate resistant plants by compensating for the lack of natural resistance. Tomato (Solanum lycopersicum) and related species offer a model in which natural and TILLING‐induced potyvirus resistance alleles may be compared. For resistance based on translation initiation factor eIF4E1, we confirm that the natural allele Sh–eIF4E1PI24–pot1, isolated from the wild tomato species Solanum habrochaites, is associated with a wide spectrum of resistance to both potato virus Y and tobacco etch virus isolates. In contrast, a null allele of the same gene, isolated through a TILLING strategy in cultivated tomato S. lycopersicum, is associated with a much narrower resistance spectrum. Introgressing the null allele into S. habrochaites did not extend its resistance spectrum, indicating that the genetic background is not responsible for the broad resistance. Instead, the different types of eIF4E1 mutations affect the levels of eIF4E2 differently, suggesting that eIF4E2 is also involved in potyvirus resistance. Indeed, combining two null mutations affecting eIF4E1 and eIF4E2 re‐establishes a wide resistance spectrum in cultivated tomato, but to the detriment of plant development. These results highlight redundancy effects within the eIF4E gene family, where regulation of expression alters susceptibility or resistance to potyviruses. For crop improvement, using loss‐of‐function alleles to generate resistance may be counter‐productive if they narrow the resistance spectrum and limit growth. It may be more effective to use alleles encoding functional variants similar to those found in natural diversity.
Metabolic Engineering of Probiotic Saccharomyces boulardii
Authors: Liu Jing-Jing , Kong In Iok , Zhang Guo-Chang , Jayakody Lahiru N. , Kim Heejin , Xia Peng-Fei , Kwak Suryang , Sung Bong Hyun , Sohn Jung-Hoon , Walukiewicz Hanna E. , Rao Christopher V. , Jin Yong-Su
Published: February 2016

Saccharomyces boulardii
is a probiotic yeast that has been used for promoting gut health as well as preventing diarrheal diseases. This yeast not only exhibits beneficial phenotypes for gut health but also can stay longer in the gut than
Saccharomyces cerevisiae
. Therefore,
S. boulardii
is an attractive host for metabolic engineering to produce biomolecules of interest in the gut. However, the lack of auxotrophic strains with defined genetic backgrounds has hampered the use of this strain for metabolic engineering. Here, we report the development of well-defined auxotrophic mutants (
leu2
,
ura3
,
his3
, and
trp1
) through clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9-based genome editing. The resulting auxotrophic mutants can be used as a host for introducing various genetic perturbations, such as overexpression or deletion of a target gene, using existing genetic tools for
S. cerevisiae
. We demonstrated the overexpression of a heterologous gene (
lacZ
), the correct localization of a target protein (red fluorescent protein) into mitochondria by using a protein localization signal, and the introduction of a heterologous metabolic pathway (xylose-assimilating pathway) in the genome of
S. boulardii
. We further demonstrated that human lysozyme, which is beneficial for human gut health, could be secreted by
S. boulardii
. Our results suggest that more sophisticated genetic perturbations to improve
S. boulardii
can be performed without using a drug resistance marker, which is a prerequisite for
in vivo
applications using engineered
S. boulardii
.
CO Metabolism in the Thermophilic Acetogen Thermoanaerobacter kivui
Authors: Weghoff Marie Charlotte , Müller Volker
Published: February 2016

The thermophilic acetogenic bacterium
Thermoanaerobacter kivui
, previously described not to use carbon monoxide as a carbon and energy source, was adapted to grow on CO. This was achieved by using a preculture grown on H
2
plus CO
2
and by increasing the CO concentration in small, 10% increments.
T. kivui
was finally able to grow within a 100% CO atmosphere. Growth on CO was found in complex and mineral media, and vitamins were not required. Carbon monoxide consumption was accompanied by acetate and hydrogen production. Cells also grew on synthesis gas (syngas) with the simultaneous use of CO and H
2
coupled to acetate production. CO oxidation in resting cells was coupled to hydrogen and acetate production and accompanied by the synthesis of ATP. A protonophore abolished ATP synthesis but stimulated H
2
production, which is consistent with a chemiosmotic mechanism of ATP synthesis. Hydrogenase activity was highest in crude extracts of CO-grown cells, and carbon monoxide dehydrogenase (CODH) activity was highest in H
2
-plus-CO
2
- or CO-grown cells. The genome of
T. kivui
harbors two CODH gene clusters, and both CODH proteins were present in crude extracts, but one CODH was more prevalent in crude extracts from CO-grown cells.
Ultrastructural Variability of the Exosporium Layer of Clostridium difficile Spores
Authors: Pizarro-Guajardo Marjorie , Calderón-Romero Paulina , Castro-Córdova Pablo , Mora-Uribe Paola , Paredes-Sabja Daniel
Published: February 2016

The anaerobic sporeformer
Clostridium difficile
is the leading cause of nosocomial antibiotic-associated diarrhea in developed and developing countries. The metabolically dormant spore form is considered the transmission, infectious, and persistent morphotype, and the outermost exosporium layer is likely to play a major role in spore-host interactions during the first contact of
C. difficile
spores with the host and for spore persistence during recurrent episodes of infection. Although some studies on the biology of the exosporium have been conducted (J. Barra-Carrasco et al., J Bacteriol 195:3863–3875, 2013,
http://dx.doi.org/10.1128/JB.00369-13
; J. Phetcharaburanin et al., Mol Microbiol 92:1025–1038, 2014,
http://dx.doi.org/10.1111/mmi.12611
), there is a lack of information on the ultrastructural variability and stability of this layer. In this work, using transmission electron micrographs, we analyzed the variability of the spore's outermost layers in various strains and found distinctive variability in the ultrastructural morphotype of the exosporium within and between strains. Through transmission electron micrographs, we observed that although this layer was stable during spore purification, it was partially lost after 6 months of storage at room temperature. These observations were confirmed by indirect immunofluorescence microscopy, where a significant decrease in the levels of two exosporium markers, the N-terminal domain of BclA1 and CdeC, was observed. It is also noteworthy that the presence of the exosporium marker CdeC on spores obtained from
C. difficile
biofilms depended on the biofilm culture conditions and the strain used. Collectively, these results provide information on the heterogeneity and stability of the exosporium surface of
C. difficile
spores. These findings have direct implications and should be considered in the development of novel methods to diagnose and/or remove
C. difficile
spores by using exosporium proteins as targets.